E-Pub Ahead of Schedule: Bentham Science Publishers are pleased to offer electronic publication of accepted papers prior to scheduled publication. These peer-reviewed papers can be cited using the date of access and the unique DOI number. Any final changes in manuscripts will be made at the time of print publication and will be reflected in the final electronic version of the issue. Articles ahead of schedule may be ordered by pay-per-view at the relevant links by each article stated via the E-Pub Ahead of Schedule.

Disclaimer: Articles appearing in E-Pub Ahead-of-Schedule sections have been peer-reviewed and accepted for publication in this journal and posted online before scheduled publication. Articles appearing here may contain statements, opinions, and information that have errors in facts, figures, or interpretation. Accordingly, Bentham Science Publishers, the editors and authors and their respective employees are not responsible or liable for the use of any such inaccurate or misleading data, opinion or information contained of articles in the E-Pub Ahead-of- Schedule.

Editorial: New methodology and approaches to intracellular lipid transport in atherosclerosis and cardiovascular disease
Professor Basil D Roufogalis and Doctor Thomas Grewal
[PMID: 21470126 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00147]


Methodologies for investigating natural medicines for the treatment of nonalcoholic fatty liver disease (NAFLD)
Moon Sun Kim, Sidney Kung, Thomas Grewal, Basil D Roufogalis
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21470125 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00148]


Cellular cholesterol efflux pathways: Impact on Intracellular Lipid Trafficking and Methodological Considerations
Ilaria Zanotti, Elda Favari, Franco Bernini
[Abstract] [Purchase Article] [PMID: 21470124 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00149]


Following intracellular cholesterol transport by linear and non-linear optical microscopy of intrinsically fluorescent sterols
Daniel Wüstner
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21470123 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00150]


ATP Binding Cassette Transporter A1 (ABCA1) associated proteins: potential drug targets in the metabolic syndrome and atherosclerotic disease?
Christa Buechler and Sabrina Bauer
[Abstract] [Purchase Article] [PMID: 21470122 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00151]


Combined Light and Electron Microscopy using Diaminobenzidine Photooxidation to Monitor Trafficking of Lipids Derived from Lipoprotein Particles
Clemens Röhrl, Claudia Meisslitzer-Ruppitsch, Robert Bittman, Zaiguo Li, Georg Pabst, Ruth Prassl, Witta Strobl, Josef Neumüller, Adolf Ellinger, Margit Pavelka and Herbert Stangl
[Abstract] [Purchase Article] [PMID: 21470121 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00152]


Exploring sterol and lipid biology in the functional genomic era.
Mathew Traini and Wendy Jessup
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21470120 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00153]


Signal transduction pathways provide opportunities to enhance HDL and apoAI-dependent reverse cholesterol transport
Vishwaroop Mulay, Peta Wood, Carles Rentero, Carlos Enrich and Thomas Grewal
[Abstract] [Purchase Article] [PMID: 21470119 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00154]


Nanocrystals, a new tool to study lipoprotein metabolism and atherosclerosis
Joerg Heeren and Oliver Bruns
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21470118 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00155]


Novel aspects of apoptosis modulating drugs
Matthias Ocker
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605077 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00182]


New Drugs, Old Fashioned Ways: Er Stress Induced Cell Death
Pietro Di Fazio, Matthias Ocker, Roberta Montalbano
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605076 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00183]


Biomarkers for hepatocellular apoptosis in the management of liver diseases
Marie V. St-Pierre, Jean-François Dufour
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605075 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00184]


Current Status of Therapeutic Targeting of Developmental Signalling Pathways in Oncology
Tobias Kiesslich, Frieder Berr, Beate Alinger, Ralf Kemmerling, Martin Pichler, Matthias ocker, Daniel Neureiter
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605074 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00185]


Therapeutic targeting of apoptotic pathways: Novel aspects in pancreatic cancer
A. Neesse, T.M. Gress and P. Michl
[Abstract] [Purchase Article] [PMID: 21605073 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00186]


The therapeutic potential of RNA interference: novel approaches for cancer treatment
Kinya Okamoto, Yoshikazu Murawaki
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605072 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00187]


Apoptotic potency of angiostatic compounds in the treatment of cancer
Esther Raskopf, Tilman Sauerbruch, Volker Schmitz
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605071 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00188]


Combination of hypoxia and RNA-interference targeting VEGF induces apoptosis in hepatoma cells via autocrine mechanisms
Esther Raskopf, Annabelle Vogt, Georges Decker, Sarah Hirt,  Katjana Daskalow, Thorsten Cramer, Jens Standop, Maria-Angeles Gonzalez-Carmona, Tilman Sauerbruch, Volker Schmitz
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605070 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00189]


Apoptosis signalling activated by TNF in the lower gastrointestinal tract - Review
Natalya Benderska, Saritha Chakilam, Manuela Hugle, Jelena Ivanovska, Muktheshwar Gandesiri, Jan Schulze-Lührmann, Khuloud Bajbouj, Ronald Croner, Regine Schneider-Stock
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605069 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00190]


The Tumor Stroma as Mediator of Drug Resistance - A Potential Target to Improve Cancer Therapy?
Susanne Sebens and Heiner Schäfer
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605068 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00191]


Glycobiology in malignant gliomas: expression and functions of galectins and possible therapeutic options
Herwig M. Strik, Malgorzata Kolodziej, Wolfgang Oertel, Jörg Bäsecke
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21605067 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00192]


Editorial: Frontiers in Ovarian Stimulation
Gian Mario Tiboni
[PMID: 21658001 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00195]


ESR1, ESR2 and FSH receptor gene polymorphisms in combination: a useful genetic tool for the prediction of poor responders
Anagnostou E, Mavrogianni D, Theofanakis Ch, Drakakis P, Bletsa R, Demirol A, Gurgan T, Antsaklis A, Loutradis D
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21658000 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00196]


present and future of recombinant gonadotropins in reproductive medicine
Vincenzo De Leo, Maria Concetta Musacchio, Alessandra Di Sabatino, Claudia Tosti, Giuseppe Morgante, and Felice Petraglia
[Abstract] [Purchase Article] [PMID: 21657999 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00197]


Ovarian stimulation: today and tomorrow
Fatemi H. M,  Blockeel C,  Devroey P
[Abstract] [Purchase Article] [PMID: 21657998 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00198]


Significantly lower pregnancy rates in the presence of progesterone elevation in patients treated with GnRH antagonists and gonadotrophins: A systematic review and meta-analysis
E.M. Kolibianakis,  C.A. Venetis,  J  Bontis,  B.C. Tarlatzis
[Abstract] [Purchase Article] [PMID: 21657997 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00199]


Possibilities and limits of ovarian reserve testing in ART
Antonio La Marca, Cindy Argento, Giovanna Sighinolfi, Valentina Grisendi, Marilena Carbone, Giovanni D’Ippolito, Alfredo Carducci Artenisio, Gaspare Stabile, Annibale Volpe
[Abstract] [Purchase Article] [PMID: 21657996 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00200]


Genetic profile of SNP(s) and ovulation induction
Loutradis D, Theofanakis Ch, Anagnostou E, Mavrogianni D, Partsinevelos GA
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21657995 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00201]


Modeling Follicle Stimulating Hormone Levels in Serum for Controlled Ovarian Hyperstimulation I: Comparing Gonadotropin Products
David H. McCulloh, Priya Maseelall, Jose M. Colon and Peter G. McGovern, David H. McCulloh
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21657994 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00202]


Modeling Follicle Stimulating Hormone (FSH) Levels in Serum for Controlled Ovarian Hyperstimulation II: The Underlying Mechanisms
David H. McCulloh, Jose M. Colon  and Peter G. McGovern
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21657993 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00203]


Modeling Follicle Stimulating Hormone Levels in Serum for Controlled Ovarian Hyperstimulation III: Improved Gonadotropin Administration
David H. McCulloh, Jose M. Colon  and Peter G. McGovern
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21657992 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00204]


The Role of LH in Ovarian Stimulation
Elkin Muñoz, Ernesto Bosch,  Iria Fernández,  Susana Portela,  Ginna Ortiz, José Remohí, Antonio Pellicer
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21657991 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00205]


Mining the Adenovirus "Virome" for Systemic Oncolytics
Michael A. Barry, Eric A. Weaver, and Christopher Y. Chen
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740366 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00206]


Targeted and armed oncolytic poxviruses for cancer: the lead example of JX-594
Caroline J Breitbach, Steve H Thorne, John C Bell, David H Kirn
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740365 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00207]


Recent Clinical Experience With Oncolytic Viruses
O. G. Donnelly, F. Errington-Mais, R. Prestwich, K. Harrington, H. Pandha, R. Vile and A.A. Melcher
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740364 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00208]


Pro-oncogenic cell signaling machinery as a target for oncolytic viruses
Emma Borrego-Diaz, Rajesh Mathew, Dana Hawkinson, Tuba Esfandyari, Zhengian Liu, Patrick W Lee, Faris Farassati
[Abstract] [Purchase Article] [PMID: 21740363 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00209]


Synergistic Interaction of Telomerase-Specific Oncolytic Virotherapy and Chemotherapeutic Agents for Human Cancer
Toshiyoshi Fujiwara, Shunsuke Kagawa and Hiroshi Tazawa
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740362 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00210]


Attenuated oncolytic Measles Virus strains as cancer therapeutics
P. Msaouel, I.D. Iankov, A. Dispenzieri, E. Galanis
[Abstract] [Purchase Article] [PMID: 21740361 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00211]


Targeted oncolytic herpes simplex viruses for aggressive cancers
Jennifer wong, cleo lee, kevin zhang, paul s. Rennie, william jia
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740360 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00212]


Oncolytic HSV-1 Virotherapy: Clinical Experience and Opportunities for Progress
Balveen Kaur, E. Antonio Chiocca and Timothy P Cripe
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740359 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00213]


Strategy of Cancer Targeting Gene-Viro-Therapy (CTGVT) — a trend in both cancer gene therapy and cancer virotherapy
Xin Yuan Liu, Hua Guang Li, Dong Qin Yang, Jin Fa Gu
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740358 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00214]


Clinical Development of Oncolytic viruses in China
Min Liang
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740357 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00215]


The Molecular Basis Of Herpesviruses As Oncolytic Agents
Laura Menotti, Gabriella Campadelli-Fiume, Patrizia Nanni, Pier Luigi Lollini and  Carla De Giovanni
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740356 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00217]


Oncolytic Viruses for Induction of Anti-tumor Immunity
Alex W. Tong, Neil Senzer, Vincenzo Cerullo, Nancy Smyth Templeton, Akseli Hemminki, and John Nemunaitis
[Abstract] [Purchase Article] [PMID: 21740355 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00218]


Bugs and Drugs: Oncolytic Virotherapy in Combination with Chemotherapy
Sonia Tusell Wennier, Jia Liu and Grant McFadden
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740354 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00219]


Fusogenic Oncolytic Herpes Simplex Viruses as a Potent and Personalized Cancer Vaccine
Qi-Xiang Li, Guohong Liu and Xiaoliu Zhang
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740353 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00220]


Editorial: Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF): surgical adjuvants, preparations for in situ regenerative medicine and tools for tissue engineering
Tomasz Bielecki
[PMID: 21740380 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00221]


In search of a consensus terminology in the field of platelet concentrates for surgical use: Platelet-Rich Plasma (PRP), Platelet-Rich Fibrin  (PRF), fibrin gel polymerization and leukocytes
David M. Dohan Ehrenfest, Tomasz Bielecki, Allan Mishra, Piero Borzini, Francesco Inchingolo, Gilberto Sammartino, Lars Rasmusson and Peter A. Everts
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740379 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00222]


Platelet concentrates for topical use: bedside device and blood transfusion technology. Quality and Versatility
Piero Borzini, Valeria Balbo, Laura Mazzucco
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740378 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00223]


Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates ? An evidence-based answer comparing a Pure Platelet-Rich Plasma (P-PRP) Gel and a Leukocyte- and Platelet-Rich Fibrin (L-PRF).
David M. Dohan Ehrenfest, Tomasz Bielecki, Ryo Jimbo, Giovanni Barbé, Marco Del Corso, Francesco Inchingolo and Gilberto Sammartino
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740377 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00224]


The role of leukocytes from L-PRP/L-PRF in wound healing and immune defense: new perspectives
Tomasz Bielecki,  David M. Dohan Ehrenfest, Peter A. Everts and Andrzej Wiczkowski
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740376 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00225]


Is the use of autologous platelet-rich plasma gels in gynecologic, cardiac, and general, reconstructive surgery beneficial?
Peter A.M. Everts, Maarten M. Hoogbergen, Tjaarda A.Weber, Roger J.J. Devilee, Gust van Monftort, Ignace H.J.T. de Hingh
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740375 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00226]


Applications of leukocyte- and platelet-rich plasma (L-PRP) in trauma surgery
Ting Yuan, Shang-Chun Guo, Pei Han, Chang-Qing Zhang, Bing-Fang Zeng
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740374 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00227]


Sports Medicine Applications of Platelet Rich Plasma
Allan Mishra, Kimberly Harmon, James Woodall, and Amy Vieira
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740373 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00228]


Leukocyte- and platelet-rich fibrin (L-PRF) for long-term delivery of growth factor in rotator cuff repair: review, preliminary results and future directions
Matthias A. Zumstein, Simon Berger Martin Schober, Pascal Boileau, Richard W. Nyffeler, Michael Horn, and Clemens A. Dahinden
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740372 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00229]


Current knowledge and perspectives for the use of Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF) in oral and maxillofacial surgery. Part 1: periodontal and dentoalveolar surgery
Marco Del Corso, Alain Vervelle, Alain Simonpieri, Ryo Jimbo, rancesco Inchingolo, Gilberto Sammartino and David M. Dohan Ehrenfest
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740371 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00230]


Current knowledge and perspectives for the use of Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF) in oral and maxillofacial surgery. Part 2: bone graft, implant and reconstructive surgery
Alain Simonpieri, Marco Del Corso, Alain Vervelle, Ryo Jimbo, Francesco Inchingolo, Gilberto Sammartino and David M. Dohan Ehrenfest
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740370 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00231]


The Role of “Eye Platelet Rich Plasma” (E-Prp) for  Wound Healing in Ophthalmology
Jorge L. Alió, Francisco Arnalich-Montiel, Alejandra E. Rodriguez
[Abstract] [Purchase Article] [PMID: 21740369 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00232]


L-PRP/L-PRF in esthetic plastic surgery, regenerative medicine of the skin and chronic wounds
Agata Cieslik-Bielecka, Joseph Choukroun, Guillaume Odin and David M. Dohan Ehrenfest
[Abstract] [FULL-TEXT INQUIRY] [PMID: 21740368 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00233]


Editorial: Primary sjögren’s syndrome research and therapy: has a new dawn arrived?


Wan-fai Ng
Musculoskeletal Research Group
Institute of Cellular Medicine
Newcastle University
Newcastle upon Tyne
NE2 4HH
United Kingdom
Tel: +44-191-2228125
e-mail: Wan-fai.Ng@ncl.ac.uk
[BSP/CPB/E-Pub/00247]


Ectopic Lymphoid Neogenesis And Lymphoid Chemokines In Sjogren’s Syndrome: At The Interplay Between Chronic Inflammation, Autoimmunity And Lymphomagenesis
Michele Bombardieri and Costantino Pitzalis
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00248]


Autoantibodies and Sjögren’s syndrome:a physiologist’s perspective
Peter M. Smith and Luke J Dawson
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00249]


B-Lymphocytes Govern The Pathogenesis Of Sjögren's Syndrome
Pierre Youinou, Alain Saraux and Jacques-Olivier Pers
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00250]


Biologic Therapies in Primary Sjögren's syndrome
Dr S J Bowman
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00251]


Primary Sjögren’s Syndrome : Time For Prospective Cohorts
Jacques-Eric Gottenberg and Xavier Mariette
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00252]


Current And Future Challenges In Primary Sjögren’s Syndrome
Arjan Vissink, Hendrika Bootsma, Fred KL Spijkervet, Shen Hu, David T Wong and  Cees GM Kallenberg
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00253]


Primary Sjögren’s Syndrome And The Type I Interferon System
Gunnel Nordmark, Maija-Leena Eloranta and  Lars Rönnblom
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00254]


Local Activation and Systemic Dysregulation of T Lymphocytes in Sjögren’s Syndrome
Robert Busch, Andreas V. Hadjinicolaou and Frances C. Hall
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00255]


Epigenetics and Sjögren’s syndrome
Christelle Le Dantec, Marie-Michèle Varin, Wesley H Brooks, Jacques-Olivier Pers, Pierre Youinou and Yves Renaudineau
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00256]


Editorial: The bioenergetics of cancer, the Warburg hypothesis and the mitochondrial function
Jiri Neuzil and Rafael Moreno-Sánchez
[BSP/CPB/E-Pub/00257]


Cell Hierarchy, Metabolic Flexibity And Systems Approaches To Cancer Treatment
Patries M. Herst and Michael V. Berridge
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00258]


p53 Regulation of Energy Metabolism and Mitochondria Regulation of p53 in Cancer Cells: An Insight into the Role of Manganese Superoxide Dismutase
Yulan Sun, Aaron K. Holley and Daret K. St. Clair
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00259]


Combined Chemotherapy or Biotherapy with Jasmonates: Targeting Energy Metabolism for Cancer Treatment
Uri Elia and Eliezer Flescher
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00260]


Regulation of the metabolism of polyunsaturated fatty acids and butyrate in colon cancer cells
Jirina Hofmanová, Alena Vaculová and Alois Kozubík
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00261]


Use of anti-cancer drugs, mitocans, to enhance the immune responses against tumors
Hahn, T. Polanczyk, M.J. Borodovsky, A. Ramanathapuram and L.V. Akporiaye E.T Ralph, S.J.
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00262]


Targeting the Mitochondrial Electron Transport Chain Complexes for the Induction of Apoptosis and Cancer Treatment
Jakub Rohlena, Lan-feng Dong and Jiri Neuzil
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00263]


The warburg hypothesis and the atp supply in cancer cells Is oxidative phosphorylation impaired in malignant neoplasias?
Sara Rodríguez-Enríquez, Juan Carlos Gallardo-Pérez, Alvaro Marín-Hernández and Rafael Moreno-Sánchez
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00264]


Potentiation of Anti-Cancer Treatment by Modulators of Energy Metabolism
Maria C. Shoshan
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00265]


The Potential of 11C-acetate PET for Monitoring the Fatty Acid Synthesis Pathway in Tumors
Laura M. DeFord-Watts, Akiva Mintz, and Steven J. Kridel
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00266]


Regulation of Glycolytic and Mitochondrial Metabolism by Ras
J. Chesney and S. Telang
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00267]


Editorial: The chemotherapy of parasitic diseases: macrocyclic lactones
Aránzazu González-Canga
[BSP/CPB/E-Pub/00268]


History of avermectin and ivermectin, with notes on the history of other macrocyclic lactone antiparasitic agents
William C. Campbell
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00269]


Macrocyclic Lactone Anthelmintics: Spectrum Of Activity And Mechanism Of Action
Timothy G. Geary and Yovany Moreno
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00270]


Resistance to Macrocyclic Lactones
Adrian J Wolstenholme and Ray M. Kaplan
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00271]


Pharmacokinetic features of the antiparasitic Macrocyclic Lactones
Quintin A McKellar and Cengiz Gokbulut
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00272]


Macrocyclic lactones and cellular transport-related drug interactions -A perspective from in vitro assays to nematode control in the field-
Lifschitz, A. Ballent, M. Lanusse, C.
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00273]


Extra-Label Use of Ivermectin in Some Minor Ruminant Species: Pharmacokinetic Aspects
González-Canga, A. Belmar-Liberato, R. Escribano, M.
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00274]


Recent developments in the analysis of avermectin and milbemycin residues in food safety and the environment
Martin Danaher, Wolfgang Radeck, Lucija Kolar, Jemma Keegan and Vesna Cerkvenik-Flajs
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00275]


Toxicity in Animals: Target Species
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00276]
K N Woodward


Treatment of MDR1 mutant dogs with macrocyclic lactones
Joachim Geyer and Christina Janko
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00277]


Ivermectin Residue Depletion in Food Producing Species and its Presence in Animal Foodstuffs With a View to Human Safety
Escribano, M. San Andrés, M.I. de Lucas, J.J. González-Canga, A.
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00278]


Acute Human Toxicity of Macrocyclic Lactones
Chen-Chang Yang
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00279]


A review on the toxicity and non-target effects of macrocyclic lactones in terrestrial and aquatic environment
Jean-Pierre Lumaret, Faiek Errouissi, Kevin Floate, Jörg Römbke, Keith Wardhaugh
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00280]


Use of Macrocyclic Lactones in Cattle in the USA
Lora R. Ballweber and Laurie A. Baeten
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00281]


Macrocyclic Lactones for Parasite Control in Equids
E. T. Lyons and S. C. Tolliver
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00282]


Macrocyclic Lactones in the Treatment and Control of Parasitism in Small Companion Animals
Thomas J. Nolan and James B. Lok
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00283]


Avermectin Use In Aquaculture
Tor E. Horsberg
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00284]


Ivermectin in human medicine, an overview of the current status of its clinical applications
González, P. González, F.A. Ueno, K.
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00285]


Beyond providing drugs, the mectizan ® donation creating paradigm shifts in health care and health systems strengthening
Dr Adrian Hopkins
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00285]


Editorial: Current optical procedures used in cell biology

Yasutomo Nomura
Department of Systems Life Engineering
Maebashi Institute of Technology
460-1 Kamisadori
Maebashi, 371-0816
JAPAN
E-mail: ynomura@maebashi-it.ac.jp

Masataka Kinjo
Laboratory of Molecular Cell Dynamics
Faculty of Advanced Life Science
Hokkaido University
Sapporo, 001-0021
JAPAN
kinjo@sci.hokudai.ac.jp
[BSP/CPB/E-Pub/00286]


Image analysis of colocalization of nuclear DNA and GFP labelled HIF-1α in stable transformants
Takehito Goto, Michihiko Sato, Eiji Takahashi and Yasutomo Nomura
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00287]


The reaction mechanism of calcium-activated photoprotein bioluminescence
Takashi Hirano
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00288]


Low-Coherence Dynamic Light Scattering and its Potential for Measuring Cell Dynamics
Katsuhiro Ishii
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00289]


A Novel Molecular Design Strategy For Efficient Two-Photon Absorption Materials
Jun Kawamata and Yasutaka Suzuki
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00290]


Magnetically-Modulated Atomic Force Microscopy for Analysis of Soft Matter Systems
Masami Kageshima
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00291]


Single Molecular observation of DNA and DNA Complexes by Atomic Force Microscopy
Takuya Matsumoto
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00292]


Effect of Isoproterenol on Local Contractile Behaviors of Rat Cardiomyocytes Measured by Atomic Force Microscopy
Takaharu Okajima
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00293]


Current Research on Protein-Protein Interactions Among Auxin-Signaling Factors in Regulation of Plant Growth and Development
Hideki Muto and Masataka Kinjo
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00294]


Use of Carbohydrate-Conjugated Nanoparticles for an Integrated Approach to Functional Imaging of Glycans and Understanding of Their Molecular Mechanisms
Noriko Nagahori, Makiyo Uchida, Masataka Kinjo and Tadashi Yamashita
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00295]


Direct Quantification of Mitochondria and Mitochondrial DNA Dynamics
Dr. Yasutomo Nomura
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00296]


Atomic Force Microscopy for the Examination of Single Cell Rheology
Takaharu Okajima
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00297]


Current Status and Future Prospects for Research on Tyrosine Sulfation
Nobuya Sasaki
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00298]


Synthesis and Application of Visible Light Sensitive Azobenzene
Shinjiro Sawada, Nobuo Kato and Kunihiro Kaihatsu
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00299]


Fluorescence Imaging of Mitochondria in Living Cells Using a Novel Fluorene Derivative with a Large Two-Photon Absorption Cross-Section
Seiji Tani, Kenta Nakagawa, Takuya Honda, Hiromasa Saito, Yasutaka Suzuki, Jun Kawamata, Makiyo Uchida, Akira Sasaki and Masataka Kinjo
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/0300]


Highly Controllable Optical Tweezers Using Dynamic Electronic Holograms
Johtaro Yamamoto and Toshiaki Iwai
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00301]


Recent Advances in the Study of Glycosphingolipids
Tadashi Yamashita
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00302]


Editorial: Novel high through-put approach and molecular modelling in drug discovery from natural products and clinical implications
Chun Guang Li
[BSP/CPB/E-Pub/00303]


In silico search for drug targets of natural compounds
Lixia Yao
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00304]


Pharmacophore, QSAR, and Binding Mode Studies of Substrates ofHuman Cytochrome P450 2D6 (CYP2D6)using Molecular Docking and Virtual Mutations and an Application to Chinese Herbal Medicine Screening
Sui-Lin Mo, Wei-Feng Liu, Chun-Guang Li, Hai-Bin Luo, Rongshi Li and Shu-Feng Zhou
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00305]


Integrated Analysis on the Physicochemical Properties of Dihydropyridine Calcium Channel Blockers in Grapefruit Juice Interactions
Yoshihiro Uesawa, Takahiro Takeuchi and Kiminori Mohri
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00306]


Herb-drug interactions: methods to identify potential influence of genetic variations in genes encoding drug metabolizing enzymes and drug transporters Running title: Herb-drug interaction pharmacogenetics
M. Hu, D. Q. Wang, Y. J. Xiao, V. W. L. Mak and B. Tomlinson
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00307]


Editorial: The Era Of -Omics
David M. Pereira
[BSP/CPB/E-Pub/00308]


Soy Isoflavones and Cardiovascular Disease Epidemiological, clinical and -omics perspectives
Gil-Izquierdo, A. Peñalvo, J.L Gil, J.I. Medina, S. Horcajada, M.N. Lafay, S. Silberberg, M. Llorach, R. Zafrilla, P. García-Mora and  P. Ferreres, F
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00309]


Plant secondary metabolites in cancer chemotherapy: where are we?
David M. Pereira, Patrícia Valentão, Georgina Correia-da-Silva, Natércia Teixeira and Paula B. Andrade
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00310]


Genomics and Cancer Drug Resistance
António S. Rodrigues, Joana Dinis, Marta Gromicho, Célia Martins, António Laires and José Rueff
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00311]


Genomics and Pharmacogenomics of Brain Disorders
Ramón Cacabelos, Rocío Martínez-Bouza, Juan Carlos Carril, Lucía Fernández-Novoa, Valter Lombardi, Iván Carrera, Lola Corzo and Adam McKay
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00312]


Non-alcoholic steatohepatitis: new insights from OMICS studies
Martel Cécile, Degli Esposti Davide, Bouchet Antonin, Brenner Catherine and Lemoine Antoinette BRENNER Catherine
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00313]


Application of lipidomics to assess lipogenesis in drug development and pre-clinical trials
Antonin Lamazière, Claude Wolf and Peter J. Quinn
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00314]


Intestinal MicrobiOMICS to Define Health and Disease in Human and Mice
Matteo Serino, Chantal Chabo and Remy Burcelin
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00315]


Editorial: New treatment strategies in schizophrenia beyond dopamine: Glutamatergic neurotransmission and more

Christoph Mulert, MD
Department of Psychiatry
UKE Hamburg
Martinistraße 52, 20246 Hamburg, Germany.
Email: cmulert@uke.de
phone: +49/(0)40/7410-59521
fax:  +49/(0)40/7410-59805
[BSP/CPB/E-Pub/00316]


The glutamate hypothesis of schizophrenia: Neuroimaging and drug development
Alice Egerton and James M. Stone
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00317]


Selective activation of muscarinic acetylcholine receptors for the treatment of schizophrenia
Brian Dean
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00318]


Current progress in the genetic research of schizophrenia: relevance for drug discovery?
Dan Rujescu, Just Genius, Jens Benninghoff and Ina Giegling
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00319]


Serotonin Receptors as Targets for Drugs Useful to Treat Psychosis  and Cognitive Impairment in Schizophrenia
H. Y. Meltzer, B. W. Massey and M. Horiguchi
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00320]


Treatment of Cognitive Dysfunction in Schizophrenia
H.M. Ibrahim  and C.A. Tamminga
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00321]


Disturbed function of GABAergic interneurons in schizophrenia: Relevance for medical treatment?
J. Genius, I. Giegling, J. Benninghoff and D. Rujescu
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00322]


Metabotropic glutamate receptors as targets for novel antipsychotic treatments
L.J. Gray, A.J. Hannan and X. Zhang
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00323]


Subjective well-being of patient with schizophrenia as a target of drug treatment
E. Karamatskos, C. Mulert, M. Lambert and D. Naber
[Abstract] [Purchase Article] [BSP/CPB/E-Pub/00324]


Immunological Treatment Options for Schizophrenia
Norbert Müller and Markus J. Schwarz
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00325]


From Glutamatergic dysfunction to cognitive impairment: Boundaries in the therapeutic of the schizophrenia
Gaspar PA, Bustamante ML, Rojo LE and Martinez A
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00326]


Pharmacotherapy of schizophrenic patients: Achievements, unsolved needs, future research necessities
Hans-Jürgen Möller
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00327]


Recent advances in targeting the ionotropic glutamate receptors in treating schizophrenia
Robert E. McCullumsmith, John Hammond, Adam Funk and James H. Meador-Woodruff
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00328]


Functional Magnetic Resonance Imaging and treatment strategies in schizophrenia
Susanne Karch, Oliver Pogarell and Christoph Mulert
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/0029]


DTNBP1 (Dysbindin) Gene Variants: In vivo evidence for effects on hippocampal glutamate status
C Wirth, F Schubert, M Lautenschlager, R Brühl, A Klär, T Majic, UE Lang, A Ehrlich, G Winterer, T Sander, M Schouler-Ocak and J Gallinat
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00330]


Innovative treatment approaches in schizophrenia enhancing neuroplasticity: Aerobic exercise, erythropoetin and repetitive transcranial magnetic stimulation
Thomas Wobrock,. Alkomiet Hasan and Peter Falkai
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00331]


Altered Cortical GABA Neurotransmission in Schizophrenia: Insights into Novel Therapeutic Strategies
Ana Stan and David A. Lewis
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00332]


Editorial: Imaging in the age of molecular medicine

Professor Dr. Fabian Kiessling
Chair of Experimental Molecular Imaging
University of Aachen (RWTH)
Pauwelsstraße 20
D-52074 Aachen
Phone: +49 (0) 241 8080116
Fax: +49 (0) 241 803380116
E-Mail: fkiessling@ukaachen.de
[BSP/CPB/E-Pub/00333]


Investigation of Superparamagnetic Iron Oxide Nanoparticles for MR-Visualization of Surgical Implants
Slabu, G. Güntherodt, T. Schmitz-Rode, M. Hodenius, N. Krämer, H. Donker, G.A. Krombach, J.Otto, U. Klinge and M. Baumann
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00334]


Monitoring Molecular, Functional and Morphologic Aspects of Bone Metastases Using Non-Invasive Imaging
Tobias Bäuerle, Dorde Komljenovic and Wolfhard Semmler
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00335]


Imaging of hypoxia using PET and MRI
Gaertner FC, Souvatzoglou M, Brix G and Beer AJ
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00336]


Cellular and Subcellular Imaging in Live Mice Using Fluorescent Proteins
Robert M. Hoffman
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00337]


Theranostic systems and strategies for monitoring nanomedicine-mediated drug targeting
Sijumon Kunjachan, Jabadurai Jayapaul, Marianne E. Mertens, Gert Storm, Fabian Kiessling and Twan Lammers
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00338]


Imaging in the age of molecular medicine: Monitoring of anti-angiogenic treatments
W. Lederle, M. Palmowski and F. Kiessling
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00339]


Translational Optical Imaging in Diagnosis and Treatment of Cancer
Stijn Keereweer, Merlijn Hutteman, Jeroen D.F. Kerrebijn, Cornelis J.H. van de Velde, Alexander L. Vahrmeijer and Clemens W.G.M. Löwik
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00340]


Apoptosis imaging to monitor cancer therapy: The road to fast treatment evaluation?
Marijke De Saint-Hubert, Matthias Bauwens, Alfons Verbruggen and Felix M. Mottaghy
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00341]


Deep Tissue Optical and Optoacoustic Molecular Imaging Technologies for Small Animal Research and Drug Discovery
Daniel Razansky, Nikos Deliolanis, Claudio Vinegoni and Vasilis Ntziachristos
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00342]


Enzymatically Activatable Diagnostic Probes
Thorek DLJ and Grimm J
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00343]


Editorial:Carbon Monoxide – A Toxic Gas on the Edge to Clinical Application

Rene Schmidt
Department of Anesthesiology and Critical Care Medicine
University Medical Center, Hugstetter Strasse 55
D-79106 Freiburg im Breisgau
Germany
Phone: +49 761 270 23060
Fax: +49 761 270 23960
E-mail: rene.schmidt@uniklinik-freiburg.de
[BSP/CPB/E-Pub/00344]


Carbon monoxide, a Two-Face for the protection of the liver
Florian Bösch and Tung Yu Tsui
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00345]


Role of Carbon monoxide in vascular diseases
Ignazio Barbagallo, Giuseppina Marrazzo, Alessandro Frigiola, Agata Zappala and Giovanni Li Volti
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00346]


Role of Carbon Monoxide in Kidney Function: Is a little Carbon Monoxide Good for the Kidney?
Eva Csongradi, Luis A. Juncos, Heather A. Drummond, Trinity Vera and David E. Stec
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00347]


Heme Oxygenase-1/CO as Protective Mediators in Cigarette Smoke-Induced Lung Cell Injury and Chronic Obstructive Pulmonary Disease
Tamás Dolinay, Augustine M. K. Choi and Stefan W. Ryter
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00348]


Carbon Monoxide In Acute Lung Injury
Simone Faller and Alexander Hoetzel
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00349]


Cytoprotection by Inhaled Carbon Monoxide before Cardiopulmonary Bypass in Preclinical Models
Torsten Loop, Christian Schlensak and Ulrich Goebel
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00350]


Application of Carbon Monoxide for Transplantation
Atsunori Nakao and Yoshiya Toyoda
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00351]


Carbon Monoxide – Toxicity of Low-Dose Application
Rene Schmidt, Helen Ryan and Alexander Hoetzel
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00352]


Carbon Monoxide and the Pancreas
Christian I. Schwer
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00353]


Antibacterial Effects of Carbon Monoxide
Jayne Louise Wilson, Helen E Jesse, Robert K Poole and Kelly S Davidge
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00354]


Reducing the development of antibiotic resistance in critical care units
Marjolein Deege and David L. Paterson
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00355]


A novel lily-of-the-valley fragrance contrast agent for magnetic resonance and fluorescence imaging of prostate cancer cells
Alexander Sturzu, Hartmut Echner, Uwe Klose, Sumbla Sheikh, Thomas Nägele, Christian Schwentner, Ulrike Ernemann and Stefan Heckl
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00356]


Editorial: Non-invasive delivery of iRNAs, proteins, peptides, cytokines and nanoparticles

Bingmei M. Fu
Department of Biomedical Engineering
The City College of the City University of New York
160 Convent Ave.
New York, NY 10031
(212)-650-7531 (O)
(212)-650-6727 (F)
fu@ccny.cuny.edu
http://bme.ccny.cuny.edu/people/faculty/bfu
[BSP/CPB/E-Pub/00357]


Pharmacokinetics/Pharmacodynamics Model-Supported Early Drug Development
Bin Chen, Jennifer Q. Dong, Wei-Jian Pan and Ana Ruiz
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00358]


Ultrasound-Induced Blood-Brain Barrier Opening
Elisa E. Konofagou, Yao-Sheng Tung, James Choi, Thomas Deffieux, Babak Baseri and Fotios Vlachos
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00359]


Experimental Methods and Transport Models for Drug Delivery across the Blood-Brain Barrier
Bingmei M Fu
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00360]


Nanocarriers for the Simultaneous Co-Delivery of Therapeutic Genes and Anticancer Drugs
Nazia Choudhury and Huixin He
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00361]


Challenges and Strategies in Developing Microneedle Patches For Transdermal Delivery of Protein and Peptide Therapeutics
Fei Wu, Sixing Yang, Weien Yuan and Tuo Jin
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00362]


Release of Growth Factors, Cytokines and Therapeutic Molecules by Hyaluronan-based Hydrogels
Robert A. Peattie
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00363]


Nanocarriers to solid tumors: considerations on tumor penetration and exposure of tumor cells to therapeutic agents
Manali Bhagat, Susan Halligan and Stavroula Sofou
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00364]


Critical Issues In Delivery of RNAi Therapeutics In Vivo
Stephanie Rivera and Fan Yuan
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00365]


Editorial: Drug development in dermatology: new targets for old diseases?

Riccardo G. Borroni, M.D., Ph.D.
Molecular Diagnostics Laboratory
Fondazione IRCCS Policlinico S. Matteo
Viale C. Golgi, 19
27100 Pavia, Italy
Phone: +39 0382 501508
Fax: +39 0382 526379
e-mail: r.borroni@smatteo.pv.it
[BSP/CPB/E-Pub/00366]


Psoriasis, from pathogenesis to therapeutic strategies: IL-21 as a novel potential therapeutic target
Elisabetta Botti, Giulia Spallone, Roberta Caruso, Giovanni Monteleone, Sergio Chimenti and Antonio Costanzo
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00367]


Role of Defensins and Cathelicidin LL37 in Auto-immune and Auto-inflammatory diseases
Loredana Frasca and Roberto Lande
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00368]


New pharmaceutical concepts for sebaceous gland diseases: implementing today’s pre-clinical data into tomorrow’s daily clinical practice
Vasiliki A. Zampeli, Evgenia Makrantonaki, Thrasivoulos Tzellos and Christos C. Zouboulis
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00369]


Gene Therapy of Skin Adhesion Disorders
Alessia Cavazza and Fulvio Mavilio
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00370]


Apoptotic pathways in the pathogenesis of pemphigus: targets for new therapies
Roberta Lotti, Alessandra Marconi and Carlo Pincelli
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00371]


Editorial: Cardiovascular and metabolic diseases: from mechanisms to novel therapeutical implications

Borbély A
Division of Clinical Physiology
Institute of Cardiology
University of Debrecen
Medical and Health Science Center
Faculty of Medicine
H-4032, Debrecen
Móricz Zsigmond krt. 22.
Hungary
Tel./Fax: 0036-52-255928
E-mail: borattila@freemail.hu
[BSP/CPB/E-Pub/003702]


The role of an experimental model of atherosclerosis: apoE-knockout mice in developing new drugs against atherogenesis
Jacek Jawien
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003703]


Regeneration of Vessel Wall Functionality and Vascular Restoration Therapy with Biodegradable Stents – Current Status
Rafał Depukat, Łukasz Rzeszutko and Dariusz Dudek
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003704]


How drugs influencing central blood pressure prevent atherosclerosis complications?
Piotr Jankowski, Michel E. Safar and Kalina Kawecka-Jaszcz
[Abstract] [Purchase Article] [BSP/CPB/E-Pub/003705]


Novel therapeutic approaches in limiting oxidative stress and inflammation
Agnieszka Spychalowicz, Grzegorz Wilk, Tomasz Śliwa, Dominik Ludew and Tomasz J. Guzik
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003706]


Genetics of hypertrophic and dilated cardiomyopathy
Felix W. Friedrich and Lucie Carrier
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003707]


Fabry Disease Cardiomyopathy: from genes to clinical manifestations
Attila Béla Kertész and István Édes
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003708]


Understanding the molecular and cellular changes behind aortic valve stenosis
Inês Falcão-Pires, Cristina Gavina and Adelino F. Leite-Moreira
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003709]


Pharmacotherapy of aortic stenosis – success or failure ?
Pawel Petkow Dimitrow, Marek Jawien and Renata Rajtar-Salwa
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003710]


Left ventricular hypertrophy in isolated aortic stenosis: primetime for the ventricle
Cristina Gavina, Inês Falcão-Pires, Francisco Rocha-Gonçalves and Adelino Leite-Moreira
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003711]


How cardiomyocytes make the heart old
Zoltán Papp, Dániel Czuriga, László Balogh, Ágnes Balogh and Attila Borbély
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003712]


Alteration of the beta-adrenergic signaling pathway in human heart failure
Nazha Hamdani and Wolfgang A. Linke
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003713]


Cellular mechanisms for diastolic dysfunction in the human heart
Dániel Czuriga, Walter J. Paulus, István Czuriga, István Édes, Zoltán Papp and Attila Borbély
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003714]


Diastolic heart failure and LV dyssynchrony
Mario Kasner, Dirk Westermann, Heinz-Peter Schultheiss and Carsten Tschöpe
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003715]


Development of Stable Lyophilized Protein Drug Products
Richard L. Remmele, Sampathkumar Krishnan and William J. Callahan
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003716]


Editorial: New Frontiers in Immunology and Infectious Diseases
Stefan Borgmann
[BSP/CPB/E-Pub/003717]


Bacteria and their toxins tamed for immunotherapy
Irena Adkins, Jana Holubova, Martina Kosova and Lenka Sadilkova
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003718]


Clinical Experience with Gene Therapy and Bispecific Antibodies for T Cell-based Therapy of Cancer 
Patrick A. Baeuerle and Christian Itin
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003719]


Current Immunotherapy of Multiple Sclerosis and Future Challenges: Relevance of Immune-mediated Repair
Michael D. Carrithers
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003720]


Infections in patients with rheumatic diseases treated with TNF antagonists
Beatriz Pérez-Zafrilla, Loreto Carmona and Juan J Gómez-Reino
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003721]


Monoclonal Antibodies For The Immunotherapy Of Solid Tumours
Richard Mauerer and Rudolf Gruber
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003722]


The intrinsic apoptosis pathways as a target in anticancer therapy
Verena Jendrossek
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003723]


Recent Advances in Active Specific Cancer Vaccine Treatment for Colorectal Cancer
Kiyotaka Okuno, Fumiaki Sugiura, Kyogo Itoh, Koji Yoshida, Takuya Tsunoda and Yusuke Nakamura
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003724]


Dendritic cells: elegant arbiters in human reproduction
Sabine e. Segerer, Claudia Rennemeier, Ulrike Kaemmerer, Torsten Frambach, Johannes Dietl and Lorenz Rieger
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003725]


Editorial: Biomedical Engineering Imaging
Kerry Hourigan

Division of Biological Engineering, Monash University, Wellington Road, Clayton 3800, Australia. kerry.hourigan@monash.edu
[BSP/CPB/E-Pub/003726]


Dye Visualization – A Method for Investigating Biomechanical Flows
Thomas Leweke
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003727]


Functional Imaging to Understand Biomechanics: A Critical Tool For the Study of Biology, Pathology and the Development of Pharmacological Solutions
R. Aidan Jamison, James A. Armitage, Josie Carberry, Marcus J. Kitchen, Stuart B. Hooper and Andreas Fouras
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003728]


Patient-Specific Modeling and Multi-Scale Blood Simulation for Computational Hemodynamic Study on the Human Cerebrovascular System
Marie Oshima,  Ryo Torii,  Shigefumi Tokuda, Shigeki Yamada and Akio Koizumi
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003729]


Recent software developments and applications in functional imaging
Lingfeng Wen, Stefan Eberl, Michael Fulham and (David) Dagan Feng
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003730]


Ultrasound: Medical Imaging and Beyond
Haim Azhari
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003731]


Opto-acoustic Imaging of Drug Discovery Biomarkers
Bohumil Bednar and Vasilis Ntziachristos
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003732]


Generation of Human single-chain Antibody to the CD99 Cell Surface Determinant Specifically Recognizing Ewing’s Sarcoma Tumor Cells
Mara Gellini, Alessandro Ascione, Michela Flego, Alessandra Mallano, Maria Luisa Dupuis, Silvia Zamboni, Manuela Terrinoni, Valeria D’Alessio, Maria Cristina Manara, Katia Scotlandi, Piero Picci and Maurizio Cianfriglia
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003733]


Fluorescence Detection of MMP-9.II. Ratiometric FRET-Based Sensing With Dually Labeled Specific Peptide
Rafal Fudala,Ryan Rich,Anindita Mukerjee, Amalendu P. Ranjan,Jamboor K. Vishwanatha, Anna K. Kurdowska, Zygmunt Gryczynski, Julian Borejdo and Ignacy Gryczynski
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003734]

Editorial: “Antidotes and rescue therapies”

Richard Y. Wang, DO
National Center for Environmental Health Centers for Disease Control and Prevention
Atlanta, GA 30341 rywang@cdc.gov

Ziad N. Kazzi, MD
Department of Emergency Medicine
Emory University
Atlanta, Georgia, 30303
[BSP/CPB/E-Pub/003735]


Antidote-Control Of Aptamer Therapeutics: The Road To A Safer Class Of Drug Agents
K.M. Bompiani, R.S. Woodruff, R.C. Becker, S.M. Nimjee and B.A. Sullenger
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003736]


Black Widow Spider (Latrodectus Mactans) Antivenom In Clinical Practice
Andrew A. Monte
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003737]


Antidotes For Acute Cyanide Poisoning
Stephen W. Borron and Frederic J. Baud
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003738]


Dexrazoxane For The Prevention Of Cardiac Toxicity And Treatment Of Extravasation Injury From The Anthracycline Antibiotics
James H. Doroshow
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003739]


Calcium And Zinc Dtpa Administration For Internal Contamination With Plutonium-238 And Americium-241
Ziad N. Kazzi, Alexander Heyl and Dr. Johann Ruprecht
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003740]


Legalon® Sil: The Antidote Of Choice In Patients With Acute Hepatotoxicity From Amatoxin Poisoning
Ulrich Mengs, Ralf -Torsten Pohla and Todd Mitchellb
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003741]


Reactive Skin Decontamination Lotion (Rsdl) For The Decontamination Of Chemical Warfare Agent (Cwa) Dermal Exposure
Schwartz, Hurst, Kirk, Reedy and Braue EH Jr
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003742]


Antivenoms For Snakebite: Design, Function, And Controversies
Eric J. Lavonas
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003743]


Acetylcysteine Therapy for Acetaminophen Poisoning
Kennon Heard and Jody Green
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003744]


Editorial: Biomedical applications of molecular vibrational imaging
Yoshinori Harada

Department of Pathology and Cell Regulation
Kyoto Prefectural University of Medicine
Graduate School of Medical Science
465 Kajii-cho
Kawaramachi Hirokoji
Kamigyo-ku, Kyoto
602-8566
Japan
TEL: +81-75-251-5322
Fax: +81-75-251-5353,
E-mail: yoharada@koto.kpu-m.ac.jp
[BSP/CPB/E-Pub/003745]


Biomedical applications of molecular vibrational imaging “Metallic nanoparticles as SERS agents for biomolecular imaging”
Jun Ando and Katsumasa Fujita
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003746]


Near-infrared spectroscopy in studies of brain oxygenation
Hideo Eda
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003747]


Raman molecular imaging of cells and tissues: towards functional diagnostic imaging without labeling
Yoshinori Harada and Tetsuro Takamatsu
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003748]


Coherent anti-Stokes Raman scattering microscopy for high speed non-staining biomolecular imaging
Mamoru Hashimoto,Takeo Minamikawa and Tsutomu Araki
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003749]


Precise analysis of the autofluorescence characteristics of rat colon under UVA and violet light excitation
Keimei Nakano, Yoshinori Harada, Yoshihisa Yamaoka, Kiichiro Miyawaki, Katsuichi Imaizumi, Hideyuki Takaoka, Masaya Nakaoka, Naoki Wakabayashi, Toshikazu Yoshikawa and Tetsuro Takamatsu
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003750]


IR super-resolution microspectroscopy and its application to single cells
Makoto Sakai, Keiichi Inoue and Masaaki Fujii
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003751]


FRET Enhanced Fluorescent Nanodiamonds
Rafal Fudala, Sangram Rout, Badri P. Maliwal, T. W. Zerda, Ignacy Gryczynski, Eric Simanek, Julian Borejdo, Ryan Rich, Irina Akopova and Zygmunt Gryczynski
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003752]


Editorial: New targets and new drugs: from in silico planning to in vivo testing
Guest Editor: Giovanni Maga
[BSP/CPB/E-Pub/003753]


Allosteric Inhibitors of Bcr-Abl: Towards Novel Myristate-Pocket Binders ?
Marco Radi, Silvia Schenone and Maurizio Botta
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003754]


Therapeutic Genes for Anti-HIV/AIDS Gene Therapy
chiara bovolenta, simona porcellini and luca alberici
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003755]


Peptides as drugs: From screening to application
Ursula Dietrich, Ralf Dürr and Joachim Koch
[Abstract] [Purchase Article] [BSP/CPB/E-Pub/003756]


Capsid Assembly as a Point of Intervention for Novel Anti-viral Therapeutics
Vishwanath R. Lingappa, Clarence R. Hurt and Edward Garvey
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003757]


Poly(ADP-ribosylation) and neoplastic transformation:Effect of PARP inhibitors
Francesca Donà, Ilaria Chiodi, Cristina Belgiovine, Tatiana Raineri, Roberta Ricotti, Chiara Mondello and A. Ivana Scovassi
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003758]


X-ray Crystallography as a tool for Mechanism-of-action Studies and Drug Discovery
Eric Ennifar
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003759]


The power of enzyme kinetics in the drug development process
Alberta Samuele, Emmanuele Crespan, Anna Garbelli, Laura Bavagnoli and Giovanni Maga
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003760]


Simple and General Criterion for “In Silico” Screening of Candidate HIV Drugs
Nevena Veljkovic, Sanja Glisic, Jelena Prljic, Vladimir Perovic and Veljko Veljkovic
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003761]



Abstracts

Methodologies for investigating natural medicines for the treatment of nonalcoholic fatty liver disease (NAFLD)
Moon Sun Kim, Sidney Kung, Thomas Grewal, Basil D Roufogalis
[FULL-TEXT INQUIRY] [PMID: 21470125 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00148]

Non-alcoholic fatty liver disease (NAFLD) is emerging as a prominent condition in Western countries. In this review we describe the characteristics and current treatments of NAFLD and discuss opportunities for developing new therapeutic management approaches, with a particular emphasis on development of animal studies and in vitro assays for identification of components of natural product medicines. The main manifestation of NAFLD is hepatic lipid accumulation in the form of lipid droplets (LDs), known as hepatic steatosis (fatty liver). Current treatments for NAFLD generally aim to reduce triglyceride (TG) accumulation, often utilizing thiazolidinedines (TZDs) and fibrates, which are known to lower TG levels in hyperlipidemia, diabetes and metabolic syndrome. Both of these compounds act through activation of nuclear receptors of the Peroxisome Proliferator-Activated Receptor (PPAR) family, thereby activating genes involved in triglyceride metabolism. Thus treatment using natural PPARα and PPARγ ligands, such as polyunsaturated fatty acids (PUFA), has also been considered. Alternatively, natural medicines for the treatment of NAFLD have a long and successful history of controlling disease without prominent side effects. However, active compounds in natural medicine responsible for lowering hepatic TG levels are yet poorly characterized. This points to the need for medium-high throughput screening assays to identify active components within natural herbs. As outlined in this review, the quantification of the size and number of lipid droplets could provide an opportunity to screen compound libraries derived from natural medicine for their potential to reduce NAFLD.
[Back to top]


Cellular cholesterol efflux pathways: Impact on Intracellular Lipid Trafficking and Methodological Considerations
Ilaria Zanotti, Elda Favari, Franco Bernini
[Purchase Article] [PMID: 21470124 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00149]

Intracellular lipid homeostasis is regulated by multiple mechanisms devoted to the tight control of cholesterol levels. Cholesterol efflux to extracellular acceptors represents a cellular response to excess accumulation of lipids that occurs by both passive and active processes and was shown to exert a beneficial, antiatherosclerotic activity. Up to now 3 lipid transporters responsible for cholesterol removal from cells have been characterized: ATP Binding Cassette A1 (ABCA1), ATP Binding Cassette G1 (ABCG1) and Scavenger Receptor Class B Type I (SR-BI). These proteins widely differ in the pathway of efflux they mediate, as well as in the nature of extracellular acceptor they interact with. The experimental investigation of cholesterol efflux pathways can be efficiently performed in vitro, following precise criteria in the selection of cell types and extracellular acceptors to specifically investigate the mechanism involved. The aim of this review is to describe how lipid transporter-mediated cholesterol efflux influences the intracellular trafficking of cholesterol and to provide methodological considerations for experimental evaluation of this process.
[Back to top]


Following intracellular cholesterol transport by linear and non-linear optical microscopy of intrinsically fluorescent sterols
Daniel Wüstner
[FULL-TEXT INQUIRY] [PMID: 21470123 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00150]

Elucidation of intracellular cholesterol transport is important for understanding the molecular basis of several metabolic and neuronal diseases, like atheroclerosis or lysosomal storage disorders. Progress in this field depends crucially on the development of new technical approaches to follow the cellular movement of this essential lipid molecule. In this article, a survey of the various methods being used for analysis of sterol trafficking is given. Various classical biochemical methods are presented and their suitability for analysis of sterol trafficking is assessed. Special emphasis is on recent developments in imaging technology to follow the intracellular fate of intrinsically fluorescent sterols as faithful cholesterol markers. In particular, UV-sensitive wide field and multiphoton microscopy of the sterol dehydroergosterol, DHE, is explained and new methods of quantitative image analysis like pixel-wise bleach rate fitting and multiphoton image correlation spectroscopy are introduced. Several applications of the new technology including observation of vectorial sterol trafficking in polarized human hepatoma cells for investigation of reverse cholesterol transport are presented.
[Back to top]


ATP Binding Cassette Transporter A1 (ABCA1) associated proteins: potential drug targets in the metabolic syndrome and atherosclerotic disease?
Christa Buechler and Sabrina Bauer
[Purchase Article] [PMID: 21470122 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00151]

The metabolic syndrome is defined as a cluster of disorders including visceral obesity, insulin resistance, hypertension, hypertriglyceridemia and low HDL. Patients have an increased risk to develop atherosclerotic disease characterized by excessive macrophage cholesterol deposition in the vascular wall. HDL removes excess cellular cholesterol which is subsequently transported to the liver for biliary excretion and thereby preserves cholesterol homeostasis. Circulating HDL levels are maintained by hepatic ATP-binding cassette transporter A1 (ABCA1) which also transports peripheral cell cholesterol to extracellular lipid acceptors. Lipid export activity of ABCA1 improves pancreatic β-cell function and ameliorates insulin release. ABCA1 affects plasma membrane cholesterol distribution and formation of lipid rafts representing signalling platforms for diverse receptors including toll-like receptor 4 (TLR4). Pharmacological activation of ABCA1 pathways presumably progresses metabolic diseases, and current approaches demonstrate beneficial effects of small peptides mimicking ABCA1 ligands which stabilize ABCA1 and enhance lipid efflux similar to its physiological acceptor apolipoprotein A-I.

Research of the last decade has resulted in the identification of several ABCA1 binding proteins influencing ABCA1 signalling, stability and activity. In the current review the proteins suggested to form a complex with ABCA1 are summarized and their up to now characterized features towards ABCA1 functions are described.
[Back to top]


Combined Light and Electron Microscopy using Diaminobenzidine Photooxidation to Monitor Trafficking of Lipids Derived from Lipoprotein Particles
Clemens Röhrl, Claudia Meisslitzer-Ruppitsch, Robert Bittman, Zaiguo Li, Georg Pabst, Ruth Prassl, Witta Strobl, Josef Neumüller, Adolf Ellinger, Margit Pavelka and Herbert Stangl
[Purchase Article] [PMID: 21470121 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00152]

Diaminobenzidine (DAB) photooxidation is a method for conversion of fluorescent signals into electron-dense precipitates that are visible in the electron microscope. Recently, we have applied this method to analyze organelles involved in holo-high density lipoprotein (HDL) particle uptake at the ultrastructural level. In the present work we extended the spectrum of molecules visualized via photooxidation to monitor the uptake of HDL-derived lipids in HepG2 cells. By the combined light-electron microscopic method and with the aid of the DAB photooxidation technique, it became possible for the first time to visualize different intracellular pathways of lipoprotein particle-derived lipids and analyze the compartments involved at the ultrastructural level. HDL-Alexa 568 was used to visualize holo-HDL particle uptake. Reconstituted HDL particles containing the fluorescent cholesterol analogues Bodipy-cholesterol, Bodipy-cholesteryl oleate, or cholesteryl Bodipy-ester were used to visualize uptake of the HDL-associated sterol. In Bodipy-cholesteryl oleate and cholesteryl Bodipy-ester, the cholesterol moiety or the fatty acid moiety is fluorescently labeled, respectively; in contrast, Bodipy-cholesterol is an analogue of free cholesterol. The cellular compartments involved in their intracellular routes after uptake were analyzed in the fluorescence and electron microscope after DAB photooxidation. Bodipy-cholesterol was found to be localized in tubular endosomes and multivesicular bodies (MVBs), in the trans-Golgi network, and in stacked Golgi cisternae. In contrast, HepG2 cells incubated with HDL containing Bodipy-cholesteryl oleate or cholesteryl Bodipy-ester gave an uptake pattern comparable to that of holo-HDL particles, with MVBs being involved. Bodipy-cholesteryl oleate was also found in lysosomes. These results indicate that HDL-derived cholesterol and cholesteryl ester are transported by different intracellular pathways in HepG2 cells. Thus, the DAB photooxidation method enables the analysis of intracellular transport of lipoprotein particle-derived lipids at the light and at the ultrastructural level.
[Back to top]


Exploring sterol and lipid biology in the functional genomic era.
Mathew Traini and Wendy Jessup
[FULL-TEXT INQUIRY] [PMID: 21470120 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00153]

The growing prevalence of disorders of lipid and sterol homeostasis such as obesity and atherosclerosis in Western societies has increased demand for the development of therapies for their prevention and treatment. Crucial for this development is an understanding of the underlying cellular mechanisms which are involved in both healthy and diseased states. However, gaps remain in our knowledge of fundamental processes, such as intracellular sterol transport, lipid storage and mobilisation. Functional genomic strategies, such as genome-wide gene knockdown screens, are increasingly being exploited as powerful strategies to fill these gaps. This review discusses experimental approaches and findings in recent functional genomics studies of sterol and lipid biology, both in model organisms and human cells.
[Back to top]


Signal transduction pathways provide opportunities to enhance HDL and apoAI-dependent reverse cholesterol transport
Vishwaroop Mulay, Peta Wood, Carles Rentero, Carlos Enrich and Thomas Grewal
[Purchase Article] [PMID: 21470119 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00154]

Binding of High Density Lipoprotein (HDL) and its major apolipoprotein A-I (apoA-I) to cell surface receptors is believed to initiate a plethora of signaling cascades that promote atheroprotective cell behavior, including the removal of excess cholesterol from lipid-loaded macrophages. More specifically, HDL and apoA-I binding to scavenger receptor BI (SR-BI) and ATP-binding cassette (ABC) transporter A1 has been shown to activate protein kinase A and C (PKA, PKC), Rac/Rho GTPases, Janus Kinase 2 (JAK2), calmodulin as well as mitogen-activated protein kinases (MAPK). Some of these signaling events upregulate mobilization of cholesterol from cellular pools, while others promote efflux pathways through increased expression, stability, and cell surface localization of SR-BI and ABCA1. This review aims to summarize the current knowledge of HDL- and apoA-I -induced signal transduction pathways that are linked to cholesterol efflux and discusses the underlying mechanisms that could couple ligand binding to SR-BI and ABCA1 with signaling and cholesterol export. Additional focus is given on the potential of pharmacological intervention to modulate the activity of signaling cascades for the inhibition or regression of cholesterol accumulation in atherosclerotic lesions.
[Back to top]


Nanocrystals, a new tool to study lipoprotein metabolism and atherosclerosis
Joerg Heeren and Oliver Bruns
[FULL-TEXT INQUIRY] [PMID: 21470118 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00155]

Nanotechnology deals with structures with a maximum size of 100 nanometers and is applied in various scientific disciplines. The basis for this is its potential to create many new materials such as nanoparticles which are suitable for a vast range of applications in electronics or energy production but also in biomedicine. Nanoparticles have exceptional physical properties useful for different applications ranging from material sciences to biomedical imaging. In life sciences nanoparticles provide a novel tool to study metabolic processes such as the metabolism of lipoproteins or to non-invasively detect diseases in a very early stage. Major hallmarks of early atherosclerotic lesion formation are endothelial dysfunction and accumulation of large amounts of lipoprotein-derived cholesterol esters in macrophages within the vessel wall. Since conventional methods such as plasma marker analyses are not specific and sensitive enough to reliably assess the risk of cardiovascular events at an early stage, nanoparticles-based imaging technologies might provide a valuable tool for the non-invasive assessment of atherosclerotic lesions in the future. In this review, we will give an overview on the characteristics of modern nanoparticles and will emphasize the current studies utilizing nanoparticles for the visualization of both lipoprotein metabolism and atherosclerosis.
[Back to top]


Novel aspects of apoptosis modulating drugs
Matthias Ocker
[FULL-TEXT INQUIRY] [PMID: 21605077 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00182]


New Drugs, Old Fashioned Ways: Er Stress Induced Cell Death
Pietro Di Fazio, Matthias Ocker, Roberta Montalbano
[FULL-TEXT INQUIRY] [PMID: 21605076 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00183]

Discovery of small molecules able to induce several cellular self-killing mechanisms improved cancer therapy in the last years. Research focused on canonical apoptotic (mitochondria or death receptor related) pathways to induce cell death in several hematologic and solid malignancies, showing that treatment with different synthetic and natural compounds reactivates the cell death machinery previously silenced in resistant cancer cells. Besides the canonical apoptotic pathways, alternative pathways of cell death induction have recently been rediscovered as potential new targets for cancer therapy.

Under certain conditions, protein folding can be disturbed causing an accumulation of unfolded proteins inside the endoplasmic reticulum (ER). This situation leads to stress ER, involving the transcriptional and translational machinery to induce the expression and post-transcriptional modifications of many factors involved in ER stress response mediated cell death. In this scenario, some apoptotic players like caspase 4 or caspase 12 start to control cell fate by inducing downstream cell death proteins.  Recently, inhibitors of protein deacetylases have been demonstrated to potently induce this alternative cell death pathway and will be reviewed here.
[Back to top]


Biomarkers for hepatocellular apoptosis in the management of liver diseases
Marie V. St-Pierre, Jean-François Dufour
[FULL-TEXT INQUIRY] [PMID: 21605075 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00184]

Apoptosis is a rare event in normal hepatocytes.  However, multiple signals can trigger apoptosis in hepatocytes and it plays a role in the pathogenesis of many liver diseases.  This review summarizes the mechanisms of hepatocellular apoptosis and the importance of apoptosis in the pathological processes of liver disease.  The potential for non-invasive biomarkers of apoptosis to gauge the extent and follow the evolution of clinical disease is emphasized.
[Back to top]


Current Status of Therapeutic Targeting of Developmental Signalling Pathways in Oncology
Tobias Kiesslich, Frieder Berr, Beate Alinger, Ralf Kemmerling, Martin Pichler, Matthias ocker, Daniel Neureiter
[FULL-TEXT INQUIRY] [PMID: 21605074 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00185]

Signalling pathways such as Hedgehog (Hh), Wnt, Notch, bone morphogenetic protein (BMP) and transforming growth factor-β (TGF-β) hold a central position in regulation of vertebrate development by controlling vital processes such as migration, differentiation and proliferation. Insights into the mechanistic aspects of cancer initiation and progression have pointed to striking similarities between tumourigenesis and embryonic development. These observations can partly be explained by the fact that similar cellular signalling mechanisms are employed in both situations. This review focuses on the role and therapeutic potential of Hh, Wnt, Notch and BMP/TGF-β signalling and discusses i) their signal transduction mechanisms during development and tumourigenesis, ii) evidence of pathway activation in different types of cancers, and, iii) strategies for pharmacological targeting. Numerous studies have demonstrated a crucial role of developmental signalling in a variety of tumours, where their signalling mechanisms contribute to oncogenic properties such as tumour cell proliferation, apoptosis inhibition and / or metastatic migration. From the literature available, it is obvious that the relative importance and the oncogenic mechanisms of developmental pathways vary with the tumour type, the stage of the disease as well as the interaction with the tumour microenvironment, thus highlighting the complexity of cellular signalling strategies employed during tumourigenesis. Intensive research activities are devoted to identification of drugs that interfere with oncogenic signalling by developmental pathways. First clinical data for such compounds – e.g. GDC-0449 for the Hh pathway – are promising and indicate that targeted therapy of developmental signalling pathways has potential for future anti-cancer therapies.
[Back to top]


Therapeutic targeting of apoptotic pathways: Novel aspects in pancreatic cancer
A. Neesse, T.M. Gress and P. Michl
[Purchase Article] [PMID: 21605073 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00186]

Pancreatic cancer constitutes one of the most aggressive tumours with a 5-year survival rate of less than 5%. It is characterized by a high degree of resistance to apoptosis which is associated by high expression levels of multiple pro-survival proteins of the extrinsic and intrinsic apoptosis signalling cascades. This review focuses on current knowledge of apoptotic pathways involved in pancreatic cancer and mechanisms of resistance to apoptosis, including alterations in the death receptor and mitochondrial pathways, as well as anti-apoptotic effects of NF-kB and Akt signalling and the impact of histon-modifying enzymes such as histondeacetylases (HDAC). Furthermore, the therapeutic implications of modulating pro-survival pathways by specific inhibitors investigated in preclinical and clinical trials will be discussed.
[Back to top]


The therapeutic potential of RNA interference: novel approaches for cancer treatment
Kinya Okamoto, Yoshikazu Murawaki
[FULL-TEXT INQUIRY] [PMID: 21605072 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00187]

Exo- and endogenous RNA interference (RNAi) is a recently developed post-transcriptional gene silencing system and is regarded as one of the most effective and specific gene silencing techniques with therapeutic potential. siRNA technology is also expected to be an invaluable treatment tool for viral infections, dominant disorders, neurological disorders, and cancers. Novel and increasingly effective therapies are urgently needed as conventional radio/chemotherapy is of limited efficacy in advanced stages of some malignant diseases. In some cancers, it is known that resistance to chemotherapy is mainly attributed to increased expression of anti-apoptotic genes, e.g. members of the bcl-2 family that stabilize the mitochondrial membrane. The siRNA-mediated knockdown of bcl-2 leads to pronounced anti-tumor effects in a pancreatic cancer model, especially in combination with chemotherapy even at otherwise ineffective concentrations. The future success of this approach will depend on the development of effective, specific, and safe delivery systems.

In addition to therapeutic RNAi, endogenous RNAi processes may also contribute to cancer development. For example, miRNAs have been shown to suppress target gene expression through binding to the 3’-untranslated regions (3’-UTR) of target mRNAs. miRNAs control many functions in regard to cell viability, including differentiation, proliferation, and apoptosis, especially during cancer progression. More investigation into miRNAs will lead to new diagnostic and therapeutic approaches in the near future.
[Back to top]


Apoptotic potency of angiostatic compounds in the treatment of cancer
Esther Raskopf, Tilman Sauerbruch, Volker Schmitz
[FULL-TEXT INQUIRY] [PMID: 21605071 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00188]

When tumours outgrow their vascular supply, they become hypoxic because of nutrient deficiency. This increases the expression and secretion of proangiogenic factors, like vascular endothelial growth factor (VEGF), leading to the activation of endothelial cells. The activated endothelial cells migrate, proliferate and form new blood vessels, resulting in increased tumour growth. This process is called tumour angiogenesis. Inhibiting tumour angiogenesis and therefore tumour growth is a well known concept in the treatment of cancer, such as hepatocellular carcinoma (HCC). This can be done by endogenous angiogenesis inhibitors, like angiostatin and its derivates. These are known to affect endothelial cell functions including the induction of apoptosis. The impact of these angiostatic factors on the cell is manifold. This also applies for so called small molecules, which affect tyrosine kinases such as receptors or intracellular signal transduction proteins. Other approaches, like monoclonal antibodies, target a single molecule, mainly VEGF, to inhibit receptor-binding and downstream signal transduction. Gene silencing, mainly via RNA interference (RNAi) intervenes on RNA-level, leading to reduced gene expression and protein secretion. Due to intense research in this field, there is rising evidence that also tumour cells themselves are influenced by angiostatic treatment approaches and the underlying molecular mechanisms are more and more revealed. Here we give a (short) review regarding the pro-apoptotic potency of anti-angiogenic compounds like angiostatic molecules, sequestering antibodies, small molecules and RNAi approaches targeting endothelial and tumour cell survival to inhibit angiogenesis and tumour growth.
[Back to top]


Combination of hypoxia and RNA-interference targeting VEGF induces apoptosis in hepatoma cells via autocrine mechanisms
Esther Raskopf, Annabelle Vogt, Georges Decker, Sarah Hirt,  Katjana Daskalow, Thorsten Cramer, Jens Standop, Maria-Angeles Gonzalez-Carmona, Tilman Sauerbruch, Volker Schmitz
[FULL-TEXT INQUIRY] [PMID: 21605070 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00189]

Control of VEGF signaling is an intense objective of pre-clinical and clinical studies in HCC disease with steadily increasing clinical application. Despite its emerging role, several aspects of anti-VEGF based treatments are poorly investigated, like the impact on tumor cells themselves, such as the effect on intracellular signaling and apoptosis induction in hepatoma cells.

Effects of siRNA-VEGF on VEGF, VEGF-receptor expression and VEGF-A signaling such as AKT and JNK phosphorylation were determined under normoxic or hypoxic conditions in murine hepatoma cells. Apoptosis induction was analyzed by SubG1-fraction, JC1-staining and caspase-8 activation. VEGF receptor expression was analysed by semiquantitative real time PCR.

Independent of oxygen status, siRNA-VEGF reduced VEGF levels resulting in decreased AKT and increased JNK phosphorylation in Hepa129 cells. The VEGF-receptors neuropilin-1 (Nrp1) and neuropilin-2 (Nrp2) were downregulated following siRNA-VEGF treatment or hypoxia induction respectively. Functionally, hypoxia significantly increased the apoptosis rate (as analyzed by SubG1-fraction, JC1-staining and JNK-phosphorylation) which was further stimulated by siRNA-VEGF treatment.

Our data indicate that antitumoral efficacy of an anti-VEGF based treatment with siRNA is partly based on negative autocrine feedback mechanisms which are even enhanced under hypoxic conditions. This observation helps to understand why antitumoral efficacy can be maintained despite of counteracting stimulation of tumoral VEGF secretion due to hypoxia. The direct impact on tumor cells further underscores the attractiveness of an anti-VEGF based siRNA treatment.
[Back to top]


Apoptosis signalling activated by TNF in the lower gastrointestinal tract - Review
Natalya Benderska, Saritha Chakilam, Manuela Hugle, Jelena Ivanovska, Muktheshwar Gandesiri, Jan Schulze-Lührmann, Khuloud Bajbouj, Ronald Croner, Regine Schneider-Stock
[FULL-TEXT INQUIRY] [PMID: 21605069 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00190]

The tumor necrosis factor (TNF) gene is an immediate early gene, rapidly transcribed in a variety of cell types following exposure to a broad range of pathogens and signals of inflammation and stress. Regulation of TNF gene expression at the transcriptional level is cell type- and stimulus-specific, involving epigenetic mechanisms or miRNAs. A better knowledge of the molecular mechanisms that control TNF gene regulation and TNF signalling will provide deeper understanding of the initiation and development of apoptotic and inflammatory processes triggered by TNF cytokine in the gut. The described efforts to embed TNF in clinical treatment regiments reflect its attractive effectiveness in killing tumor cells. Whether the described strategies will achieve the success of incorporating TNF in lower gastrointestinal tract therapy for inflammatory diseases and cancer remains to be determined.
[Back to top]


The Tumor Stroma as Mediator of Drug Resistance - A Potential Target to Improve Cancer Therapy?
Susanne Sebens and Heiner Schäfer
[FULL-TEXT INQUIRY] [PMID: 21605068 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00191]

Tumors irrespective of their origin are heterogenous cellular entities whose growth and progression greatly depend on reciprocal interactions between genetically altered (neoplastic) cells and their non-neoplastic microenvironment. Thus, microenvironmental factors promote many steps in carcinogenesis, e.g. proliferation, invasion, angiogenesis, metastasis and chemoresistance. Drug resistance, either intrinsic or acquired, essentially limits the efficacy of chemotherapy in many cancer patients. To some extent, this resistance is maintained by reduced drug accumulation, alterations in drug targets and increased repair of drug-induced DNA damage. However, the pivotal mechanism by which tumor cells elude the cytotoxic effect of chemotherapeutic drugs is their efficient protection from induction and excecution of apoptosis. It is meanwhile well established, that cellular and non-cellular components of the tumoral microenvironment, e.g. myofibroblasts and extracellular matrix (ECM) proteins, respectively, contribute to the ani-apoptotic protection of tumor cells. Cellular adhesion molecules (e.g. L1CAM or CD44), chemokines (e.g. CXCL12), integrins and other ECM receptors which are involved in direct and indirect interactions between tumor cells and their microenvironment have been identified as suitable molecular targets to overcome chemoresistance. Accordingly, several therapeutic strategies based on these targets have been already elaborated and tested in preclinical and clinical studies, including inhibitors and blocking antibodies for CD44/hyaluronan, integrins, L1CAM and CXCL12. Even though these approaches turned out to be promising, the upcoming challenge will be to prove the efficacy of these strategies in improving treatment and prognosis of cancer patients.
[Back to top]


Glycobiology in malignant gliomas: expression and functions of galectins and possible therapeutic options
Herwig M. Strik, Malgorzata Kolodziej, Wolfgang Oertel, Jörg Bäsecke
[FULL-TEXT INQUIRY] [PMID: 21605067 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00192]

Malignant gliomas, the most common malignant primary brain tumors, have a deleterious clinical prognosis of approximately 12 months in unselected series. The resistance against antineoplastic therapy is apparently not only associated with a high proliferative potential, marked antiapoptotic resistance and high migratory capacity. Effective mechanisms to escape the immune response of the organism and an intense neoangiogenesis also contribute to the aggressive growth of these neoplasms.

In addition to a number of molecular mechanisms, the group of glycohydrate-binding galectins seems to contribute to the aggressive growth of malignant gliomas. Galectin-1, -3, -4 and -8 – have been shown to be overexpressed in malignant gliomas. Galectin-1 is known to be involved in glioma cell migration and possibly also in proliferation.

In this review, various aspects of glioma biology and their therapeutic relevance is discussed. The role of galectins in apoptosis-resistance, immune response and angiogenesis is discussed and explained why these molecules interesting targets of glioma therapy.
[Back to top]


ESR1, ESR2 and FSH receptor gene polymorphisms in combination: a useful genetic tool for the prediction of poor responders
Anagnostou E, Mavrogianni D, Theofanakis Ch, Drakakis P, Bletsa R, Demirol A, Gurgan T, Antsaklis A, Loutradis D
[FULL-TEXT INQUIRY] [PMID: 21658000 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00196]

Purpose: Previous studies in humans concluded that a multigenic model including specific FSHR, ESR1 and ESR2 genotype patterns may partially explain the poor response to FSH. The aim of our study is to analyse three different loci -polymorphisms in ESR1 Pvu II, ESR2 Rsa I and Ser680Asn FSH receptor gene- in a Greek population and their involvement in stimulation outcome and pregnancy rates. Methods: Each locus was studied alone, and in combination with the others. We performed both restriction fragment length polymorphism analysis and real-time polymerase chain reaction. A total of 109 normally ovulating female patients underwent IVF or ICSI. Results: Studying each locus alone, no significant results were drawn for ESR1 and ESR2 genes. Concerning the FSHR polymorphism, the women carrying the AA variant presented higher total amount of gonadotrophins used (P=0,048) and tended to have higher number of stimulation days (P=0,057). Considering the ESR1 and FSHR gene polymorphisms in combination, the TC/SA combination presents the highest number of pregnancies in poor responders group (3/4 pregnancies carried this genotype), in good responders group (4/12 pregnancies carried this genotype) and in the total population (10/26 pregnancies carried this genotype). Except the CC/AA combination, all other genotype combinations presented incidence of pregnancy, with TC/SA having the highest incidence. The CC/AA genotype presents the worst profile of ovulation induction, confirming a poor responder profile: the total amount of gonadotrophins used was highest in CC/AA group (P<0,05). The peak E2, the number of follicles and of retrieved oocytes and the pregnancy rate were significantly lower (P<0,05). This genotype combination seems to be over-presented in the poor responders group in a statistically significant way (P=0,038). Women with CC/AA combination have 1,5-2,4 times more risk to be poor responders in comparison with women that do not carry that combination. Conclusion: This study supports the hypothesis that a multigenic model, including the well studied ESR1 and FSHR genes is involved in the controlled ovarian stimulation outcome indicating that the CC/AA genotype presents the worst ovulation induction profile, while the TC/SA genotype presents the higher number of pregnancies in our population.
[Back to top]


present and future of recombinant gonadotropins in reproductive medicine
Vincenzo De Leo, Maria Concetta Musacchio, Alessandra Di Sabatino, Claudia Tosti, Giuseppe Morgante, and Felice Petraglia
[Purchase Article] [PMID: 21657999 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00197]

Pharmacological ovarian stimulation has a major role in reproductive medicine and has been used in anovulatory patients and in the induction of multifollicular development required for the procedures of assisted reproductive techniques (ART). Currently, gonadotropins are the most important tools to proceed with ovarian stimulation for all purposes, including ART and anovulation disorders, like hypogonadotropic hypogonadism and hypothalamic hypophyseal dysfunction.

Gonadotropin preparations derived from human urine have been used clinically since the early 1960s and the first urine-derived preparation containing only FSH (urofollitropin) became available in 1983. More recently, the application of recombinant DNA technology has resulted in the development of recombinant FSH produced in mammalian cells.

In the last period, LH became available by recombinant DNA technology and is now a new option for protocols of ovarian stimulation.

Treatment with gonadotropins has been shown to be effective in males affected by hypogonadotropic hypogonadism. This success has resulted in attempts to utilize FSH therapy in oligozoospermic men, aimed at obtaining a quantitative increase in sperm count.

The purpose of this review was to examine the pharmacological aspects and different clinical applications of recombinant gonadotropins  (FSH, LH, hCG) in the treatment of female infertility in all its aspects and their use also in the treatment of male infertility. This review will trace these events, from the past through to the present, and conclude with a glance towards the future.
[Back to top]


Ovarian stimulation: today and tomorrow
Fatemi H. M,  Blockeel C,  Devroey P
[Purchase Article] [PMID: 21657998 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00198]

In assisted reproductive technology, medications and ovarian stimulation play a crucial role. The availability of gonadotrophins and GnRH analogues has allowed the tailoring of several stimulation schemes.

The two most commonly used gonadotrophin forms are urinary hMG and recombinant FSH in combination with GnRH agonists or GnRH antagonists.

Cycles stimulate with recombinant FSH appear to have a higher risk of premature progesterone rise in the late follicular phase, if not triggered on time.

Recently, corifollitropin alfa, a new long acting recombinant FSH was introduced which sustain multiple follicular growth for 7 days in women undergoing ovarian stimulation using GnRH antagonists.
GnRH antagonist and agonist do have similar live birthrate. However, GnRH antagonists reduce significantly the risk of OHSS.

Moreover, with the introduction of  GnRH antagonists, it became possible to trigger ovulation with a bolus of a GnRH agonist as an alternative to hCG.

The early OHSS  is eliminated completely with the  GnRH agonist trigger. However, due to an uncorrectable luteal phase deficiency, a poor clinical outcome is present, when GnRHa is used to trigger final oocyte maturation in IVF/ICSI antagonist protocols.

Therefore, it has been suggested to cryopreserve the embryos and transfer in consecutive natural cycles Future trials should aim to eliminate OHSS and multiple pregnancy rates by performing a single stimulation in a simplified corifolitropin alfa/GnRH antagonist cycle triggered by a GnRH agonist followed by Cryo-thawed SET in consecutive natural cycles.  With this approach, the two major complications of COH for IVF could be eliminated without jeopardizing the outcome.
[Back to top]


Significantly lower pregnancy rates in the presence of progesterone elevation in patients treated with GnRH antagonists and gonadotrophins: A systematic review and meta-analysis
E.M. Kolibianakis,  C.A. Venetis,  J  Bontis,  B.C. Tarlatzis
[Purchase Article] [PMID: 21657997 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00199]

The current meta-analysis aimed to answer the following research question: is progesterone elevation on the day of hCG administration associated with the probability of clinical pregnancy in women undergoing ovarian stimulation for IVF using GnRH antagonists? A literature search in MEDLINE, EMBASE and CENTRAL electronic databases followed by extensive hand-searching from two independent reviewers was performed to identify relevant studies. Eventually five eligible studies (n=585 patients) were identified. No significant differences were present between patients with and those without progesterone elevation regarding female age, duration of stimulation and total dose of gonadotrophins required. However, patients with progesterone elevation were characterized by higher serum estradiol levels on the day of hCG administration (+956 pg/ml, 95% +248 to +1664, random effects model, p=0.008) and more COCs retrieved (+2.9, 95% CI +1.5 to +4.4, fixed effects model, p<0.001). Progesterone elevation on the day of hCG administration was associated with a significantly decreased probability of clinical pregnancy per cycle (-9%, 95% CI -17 to -2, fixed model effects, p=0.02). In conclusion, in patients treated with GnRH antagonists and gonadotrophins,  progesterone elevation on the day of hCG administration is significantly associated with a lower probability of clinical pregnancy.
[Back to top]


Possibilities and limits of ovarian reserve testing in ART
Antonio La Marca, Cindy Argento, Giovanna Sighinolfi, Valentina Grisendi, Marilena Carbone, Giovanni D’Ippolito, Alfredo Carducci Artenisio, Gaspare Stabile, Annibale Volpe
[Purchase Article] [PMID: 21657996 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00200]

Markers of ovarian reserve are associated with ovarian aging as they decline with chronologic age, and hence may predict stages of reproductive aging including the menopause transition.

Assessment of ovarian reserve include measurement of serum follicle stimulating hormone (FSH), anti-Müllerian hormone (AMH), and inhibin-B. Ultrasound determination of antral follicle count (AFC), ovarian vascularity and ovarian volume also can have a role. The clomiphene citrate challenge test (CCCT), exogenous FSH ovarian reserve test (EFORT), and GnRH-agonist stimulation test (GAST) are dynamic methods that have been used in the past to assess ovarian reserve.

In infertile women, ovarian reserve markers can be used to predict low and high oocyte yield and treatment failure in women undergoing in vitro fertilization. However the markers may have limitations when an in depth analysis of their accuracy, cost, convenience, and utility is performed As ovarian reserve markers may permit the identification of both the extremes of ovarian stimulation, a possible role for their measurement may be in the individualization of treatment strategies in order to reduce the clinical risk of ART along with optimized treatment burden. It is fundamental to clarify the cost/benefit of its use in the ovarian reserve testing  before initiation of an IVF cycle and whether the ovarian reserve markers-determined strategy of ovarian stimulation for assisted conception may be associated to improved live birth rate.
[Back to top]


Genetic profile of SNP(s) and ovulation induction
Loutradis D, Theofanakis Ch, Anagnostou E, Mavrogianni D, Partsinevelos GA
[FULL-TEXT INQUIRY] [PMID: 21657995 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00201]

Obtaining an adequate number of good quality oocytes while minimizing adverse drug reactions (ADRs) and cycle cancellation rates is considered the gold standard in controlled ovarian hyperstimulation (COH) for fertility treatment. Patients who undergo IVF/ICSI cycles tend to present with different responses to exogenous gonadotrophin administration. Research has shown that the secret probably lies in the various single nucleotide polymorhisms (SNPs) in their receptor genes. The decryption of human genome provided specialists with additional information in assessing and even predicting ovarian response to COH. In this context, the study of Pharmacogenomics, Pharmacogenetics and SNPs unravels as a promising field in optimizing fertility treatment. Several SNPs in FSH and estrogen receptor genes have been detected so far, but only three of them, one in FSH receptor and two in estrogen receptor genes have been associated with ovarian response to COH. It seems that the Asn/Ser variant of the FSH receptor functions more efficiently, while the Ser/Ser and Asn/Asn variants have a tendency to resist to FSH stimulation. With regards to estrogen receptor 1 (ESR1), the Pvull and the Xbal polymorphisms seem to be associated with differences in the response to ovarian stimulation, while the Rsal polymorphism in estrogen receptor 2 (ESR2) is currently under investigation. There exists evidence supporting the hypothesis that a set of genes, all related to the FSH hormone mechanism of action, may participate along with other factors to the control of ovarian response to FSH, thus a cautious interpretation of polymorphism detection results is considered mandatory. However, identifying potential genetic markers that could predict ovarian response and implementing them in routine screening tests for every woman entering an IVF/ICSI cycle, would be able to tailor fertility treatment to each patients needs thus maximizing the success rate and eliminating potential side-effects of fertility drugs.
[Back to top]


Modeling Follicle Stimulating Hormone Levels in Serum for Controlled Ovarian Hyperstimulation I: Comparing Gonadotropin Products
David H. McCulloh, Priya Maseelall, Jose M. Colon and Peter G. McGovern, David H. McCulloh
[FULL-TEXT INQUIRY] [PMID: 21657994 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00202]

Background: Despite the dangers of gonadotropin administration, our understanding of serum levels of gonadotropins in our patients is poor. We created a mathematical model of gonadotropin administration in order to learn about the temporal profiles of Follicle Stimulating Hormone (FSH) levels in patients related to the doses of different gonadotropin products administered.

Methods: Model parameters for each product were determined from published pharmacokinetic information available from the manufacturers.

Results: For each product, serum levels of FSH rose then fell following a single injection of FSH. When a gonadotropin dose was injected repeatedly (daily), each injection resulted in a peak level of FSH that was higher than the level attained from the previous injection. Daily FSH peaks rose asymptotically, requiring 3-10 days to approach the maximum. The maximum averaged 3.5 times the peak of the first injection. Decline of FSH following the final injection required 3-10 days to approach the baseline. Time courses and peak values of FSH using different gonadotropin preparations varied widely.

Conclusion: Knowledge of the slow changes of FSH that occur during modeled FSH administration will provide a framework for understanding the gonadotropin profiles that occur in patients during controlled ovarian hyperstimulation.
[Back to top]


Modeling Follicle Stimulating Hormone (FSH) Levels in Serum for Controlled Ovarian Hyperstimulation II: The Underlying Mechanisms
David H. McCulloh, Jose M. Colon  and Peter G. McGovern
[FULL-TEXT INQUIRY] [PMID: 21657993 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00203]

Background: Serum levels of follicle stimulating hormone (FSH) in patients change slowly during repeated daily injections of gonadotropin, requiring several days to achieve stable values. Using a numerical (computer) model we mimicked the serum levels of FSH using two kinetic constants that describe biological phenomena: diffusion of FSH from the injection site into the blood (τ_in) and disappearance of FSH from the blood by excretion/degradation (τ_out). We investigated the effects of these two parameters on the FSH profiles seen in the model.

Methods: The parameters, τ_in and τ_out, were systematically changed to determine their effects on the delay from injection to the peak following a single injection (T_max). Other values were investigated systematically during repeated daily injections. 

Results:τ The models parameters τ_in and t_out affected T_max and several features of FSH level during daily injections of gonadotropin. These included the maximum level of FSH in the serum (relative to the level attained following the first injection), the rate of rise of peaks following injections after the first, and the shortening of the delay from injection to peak. 

Conclusion:  Knowledge of τ_in and τ_out permits us to predict serum FSH levels and time course during daily injections of gonadotropin. 
[Back to top]


Modeling Follicle Stimulating Hormone Levels in Serum for Controlled Ovarian Hyperstimulation III: Improved Gonadotropin Administration
David H. McCulloh, Jose M. Colon  and Peter G. McGovern
[FULL-TEXT INQUIRY] [PMID: 21657992 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00204]

Background:  A numerical model with two pharmacokinetic parameters (the rate of diffusion into the blood (τ_in) and the rate of disappearance from the blood (τ_out)) mimicks the slow changes of follicle stimulating hormone (FSH) seen in serum. It is often desired to change serum FSH levels more rapidly. Since the pharmacokinetic parameters cannot be changed in patients, we used the model to investigate changing the amount and timing of gonadotropin administration to change FSH levels in serum more rapidly.

Methods:  Parameters defining the model were obtained as described previously [1].  The doses administered, and the timing of administration were individually specified. 

Results: The rise of serum FSH was made more rapid by administering an initial injection larger than the subsequent injections.  When a rapid decrease of serum FSH was desired, administration of the new, lower dose was delayed. 

Conclusion: Use of a model that simulates the serum levels of FSH during gonadotropin administration provided a framework for learning how to achieve the desired serum FSH levels more rapidly in patients. With knowledge of a particular patientτs τ_in and τ_out, optimal FSH administration for that patient can be determined through modeling.
[Back to top]


The Role of LH in Ovarian Stimulation
Elkin Muñoz, Ernesto Bosch,  Iria Fernández,  Susana Portela,  Ginna Ortiz, José Remohí, Antonio Pellicer
[FULL-TEXT INQUIRY] [PMID: 21657991 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00205]

LH is a glycoprotein that plays a crucial role in folliculogenesis during the natural ovarian cycles. It has the same activity and shares receptors with hCG. However the use of LH in combination with FSH in controlled ovarian stimulation remains controversial. A practical approach concerning the usefulness of LH according to the endogenous level of LH is described herein. Specific groups of patients can benefit from ovarian stimulation with LH. New applications of LH/hCG activity are also discussed.
[Back to top]


Mining the Adenovirus "Virome" for Systemic Oncolytics
Michael A. Barry, Eric A. Weaver, and Christopher Y. Chen
[FULL-TEXT INQUIRY] [PMID: 21740366 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00206]

Adenoviruses (Ads) are arguably one of the most potent viruses for in vivo gene therapy, vaccine, and oncolytic applications. The attraction for the use of Ads stems from their ability to infect a wide range of dividing and non-dividing cell types in some cases to efficiencies of nearly 100%. Additional benefits include their stability, the ability to purify the vector to concentrations of up to 10¹³ particles/ml, and the fact that viral vectors self-assemble into particles of specific size (~100 nm). The vast majority of clinical applications of Ad have utilized Ad serotype 5 (Ad5) viruses. Considering that at least half of humans are already immune to Ad5, Ad5 oncolytics may not be optimal for clinical translation. Given this and that there are 54 different serotypes of human Ads, this review considers the utility of mining these alternate Ad serotypes for viruses that can evade Ad5 immunity and kill different types of cancer.
[Back to top]


Targeted and armed oncolytic poxviruses for cancer: the lead example of JX-594
Caroline J Breitbach, Steve H Thorne, John C Bell, David H Kirn
[FULL-TEXT INQUIRY] [PMID: 21740365 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00207]

Oncolytic viruses (OVs) are designed to replicate in, and subsequently lyse cancer cells. Numerous oncolytic virus platforms are currently in development. Here we review preclinical and clinical experience with JX-594, the lead candidate from the targeted and armed oncolytic poxvirus class. JX-594 is derived from a vaccinia vaccine strain that has been engineered for 1) enhanced cancer targeting and 2) has been “armed” with the therapeutic transgene granulocyte-macrophage colony stimulating factor (GM-CSF) to stimulate anti-tumoral immunity. Poxviruses have many ideal features for use as oncolytic agents. The development of oncolytic vaccinia viruses is supported by a large safety database accumulated in the smallpox eradication program. In addition, poxviruses have evolved unique capabilities for systemic spread through the blood that can be harnessed for the treatment of metastatic disease. JX-594 demonstrates a high degree of cancer selectivity and systemic efficacy by multiple mechanisms-of-action (MOAs) in preclinical testing. Data from Phase 1 and 2 clinical trials has confirmed that these features result in potent and systemic efficacy in patients with treatment refractory metastatic cancers.
[Back to top]


Recent Clinical Experience With Oncolytic Viruses
O. G. Donnelly, F. Errington-Mais, R. Prestwich, K. Harrington, H. Pandha, R. Vile and A.A. Melcher
[FULL-TEXT INQUIRY] [PMID: 21740364 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00208]

There has been interest in using viruses to treat cancer for over a century.  Recent clinical efforts, driven on by significant preclinical advances, have focussed on the safety of using replication-competent viruses.  Recently Publish: 1 June 2011ed clinical trials of six oncolytic viruses (adenovirus, reovirus, measles, herpes simplex, Newcastle disease virus and vaccinia) have added to the accumulating data that endorse oncolytic viruses as a safe and well tolerated treatment approach.  Conclusive evidence of efficacy remains to be demonstrated, but randomised clinical trials are now underway.
[Back to top]


Pro-oncogenic cell signaling machinery as a target for oncolytic viruses
Emma Borrego-Diaz, Rajesh Mathew, Dana Hawkinson, Tuba Esfandyari, Zhengian Liu, Patrick W Lee, Faris Farassati
[Purchase Article] [PMID: 21740363 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00209]

Viruses function in close harmony with the signaling machinery of their host. Upon exposure to the cell, a battery of viral products become engaged in boosting friendly signaling elements of the host or suppressing harmful ones. The efficiency of viral replication is indeed the biological outcome of this interaction between cellular and host signaling molecules. Oncolytic viruses, natural or man-made, follow the same set of rules of engagement. Pro-oncogenic cell signaling machinery, therefore, is undoubtedly the most important area influencing the development of the next generation of effective, specific and rationally designed oncolytic viruses. Ras signaling, with its central role in what is known today as molecular oncology, is an attractive topic for studying the behavior of viruses versus cancer cells and to develop strategies to target cancer cells on the basis of such platform. This work reviews the development of oncolytic herpes viruses capable of targeting Ras signaling pathway along with a few other examples of viruses which are developed to contain specificity for certain pro-oncogenic characteristics of their host cells.        
[Back to top]


Synergistic Interaction of Telomerase-Specific Oncolytic Virotherapy and Chemotherapeutic Agents for Human Cancer
Toshiyoshi Fujiwara, Shunsuke Kagawa and Hiroshi Tazawa
[FULL-TEXT INQUIRY] [PMID: 21740362 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00210]

Replication-selective tumor-specific viruses present a novel approach for treatment of neoplastic disease. These vectors are designed to induce virus-mediated lysis of tumor cells after selective viral propagation within the tumor. Telomerase activation is considered to be a critical step in carcinogenesis through the maintenance of telomeres, and its activity correlates closely with human telomerase reverse transcriptase (hTERT) expression. We constructed an attenuated adenovirus 5 vector, in which the hTERT promoter element drives expression of E1 genes, OBP-301 (Telomelysin). Since only tumor cells that express telomerase activity would activate this promoter, the hTERT proximal promoter allows for preferential expression of viral genes in tumor cells, leading to selective viral replication and oncolytic cell death. OBP-301 alone exhibited substantial antitumor effects both in animal models and in clinical trials; data regarding combination therapy with OBP-301 and chemotherapeutic agents are preliminary but encouraging. This article reviews synergistic interaction of virotherapy and chemotherapy, and illustrates the potential application for the treatment of human cancer.
[Back to top]


Attenuated oncolytic Measles Virus strains as cancer therapeutics
P. Msaouel, I.D. Iankov, A. Dispenzieri, E. Galanis
[Purchase Article] [PMID: 21740361 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00211]

Attenuated measles virus vaccine strains have emerged as a promising oncolytic vector platform, having shown significant anti-tumor activity against a broad range of malignant neoplasms. Measles virus strains derived from the attenuated Edmonston-B (MV-Edm) vaccine lineage have been shown to selectively infect, replicate in and lyse cancer cells while causing minimal cytopathic effect on normal tissues. This review summarizes the preclinical data that led to the rapid clinical translation of oncolytic measles vaccine strains and provides an overview of early clinical data using this oncolytic platform.  Furthermore, novel approaches currently under development to further enhance the oncolytic efficacy of MV-Edm strains, including strategies to circumvent immunity or modulate immune system responses, combinatorial approaches with standard treatment modalities, virus retargeting as well as strategies for in vivo monitoring of viral replication are discussed.
[Back to top]


Targeted oncolytic herpes simplex viruses for aggressive cancers
Jennifer wong, cleo lee, kevin zhang, paul s. Rennie, william jia
[FULL-TEXT INQUIRY] [PMID: 21740360 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00212]

Herpes simplex virus (HSV) is a well-known vector that is often used for gene therapy to treat cancers. The most attractive feature of HSV is its ability to destroy tumors through a distinctive oncolytic mechanism where the virus can destroy cancer cells via cell lysis, a killing function that no anti-cancer drugs can mimic. Importantly, HSV is a safe and effective virus that can be easily manipulated to preferentially replicate in tumor cells. In the last 20 years of re-engineering efforts, a number of HSV designs, including the classical G207, have been focused on deleting viral genes in order to render the virus tumor specific. Although such designs can successfully destroy tumor xenografts in animal models, with minimal impact on normal tissues, a common trade-off is the marked attenuation of the virus. This problem is most profound in many clinical tumors, where virus dissemination is often hindered by the difficult cellular and molecular terrain of the human tumor mass. In order to harness all of HSV’s replication potential to destroy tumor cells, efforts in our lab, as well as others, last several years have been focused on engineering an oncolytic HSV to target tumor cells without deleting any viral genes, and have since generated highly tumor specific viruses including our transcriptional translational dually regulated HSV (TTDR-HSV). In this review, we will discuss the improvements associated with the newer TTDR-HSV design compared to the classical defective HSV designs such as G207 and tk- HSV.  Lastly, we will review additional cellular features of aggressive tumors, such as their immense cellular heterogeneity and volatility, which may serve to hinder the dissemintation of TTDR-HSV. The challenge for future studies would be to explore how TTDR-HSV could be redesigned and/or employed with combinatorial approaches to better target and destroy the heterogeneous and dynamic cell populations in the aggressive tumor mass.
[Back to top]


Oncolytic HSV-1 Virotherapy: Clinical Experience and Opportunities for Progress
Balveen Kaur, E. Antonio Chiocca and Timothy P Cripe
[FULL-TEXT INQUIRY] [PMID: 21740359 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00213]

Oncolytic virotherapy with mutants derived from Herpes simplex virus (HSV) type 1 exhibit significant antitumor effects in preclinical models. Several mutants have now been tested in clinical trials for a variety of cancer types, and all have been found to be safe. While there have been hints of antitumor efficacy with prolonged survival in some cases compared with historical controls, dramatic responses have been elusive. We review the clinical experience Publish: 1 June 2011ed to date and discuss some of the biologic factors that may be limiting for virus infection and spread, as well as new strategies currently under development to enhance antitumor efficacy.
[Back to top]


Strategy of Cancer Targeting Gene-Viro-Therapy (CTGVT) — a trend in both cancer gene therapy and cancer virotherapy
Xin Yuan Liu, Hua Guang Li, Dong Qin Yang, Jin Fa Gu
[FULL-TEXT INQUIRY] [PMID: 21740358 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00214]

Cancer Targeting Gene-Viro-Therapy (CTGVT) and Gene Armed Oncolytic Virus Therapy (GAOVT) both are identical by inserting an antitumor gene into an oncolytic virus. This approach has gradually become a hot topic in cancer therapy, because that CTGVT (GAOVT) has much higher antitumor than that of either gene therapy alone or oncolytic virotherapy alone. We proposed the CTGVT strategy in 1999-2001, insisted it as a long term systematic approach to be examined over 10 years and have Publish: 1 June 2011ed 58 SCI papers some in good Journals. The CD gene armed oncolytic adenovirus therapy (GAOVT) for cancer treatment with potent antitumor effect was also named in our laboratory in 2003. Several modifications to CTGVT will be carried out by our group and will be introduced briefly in this paper. Most importantly, the modifications of CTGVT usually resulted in complete eradication of xenograft tumors in nude mice. In future best antitumor drugs may emerge from the modified CTGVT strategy and not from either gene therapy or virotherapy alone.
[Back to top]


Clinical Development of Oncolytic viruses in China
Min Liang
[FULL-TEXT INQUIRY] [PMID: 21740357 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00215]

The oncolytic virus, being a promising new therapeutic strategy for cancer, has inspired a wave of recent clinical research and development in China. The first commercialized oncolytic virus, Oncorine, was approved by Chinese SFDA in November 2005 for nasopharyngeal carcinoma combined with chemotherapy. Since then, a number of oncolytic viruses have been moved into clinical trials. Among these are the armed oncolytic adenoviruses such as H103 (expressing the heat shock protein) currently has finished phase I trial, and KH901 (expressing GM-CSF) now launched in phase II trial In this review, we will discuss the current status of ongoing oncolytic virus projects being conducted at various clinical stages in China, including the preliminary market response for Oncorine after it was  launched into the Chinese market in 2006.
[Back to top]


The Molecular Basis Of Herpesviruses As Oncolytic Agents
Laura Menotti, Gabriella Campadelli-Fiume, Patrizia Nanni, Pier Luigi Lollini and  Carla De Giovanni
[FULL-TEXT INQUIRY] [PMID: 21740356 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00217]

Herpes simplex viruses (HSVs) have entered clinical trials as oncolytic agents. The following properties make them good candidates. It is a mild pathogen; drugs (Aciclovir) are available to control viral infection; the large genome is amenable to genetic engineering, they can be rendered cancer-specific by deletion of genes, envelope glycoproteins allow the insertion of heterologous ligands to achieve modification of the natural tropism. Genetically modified HSVs have  been thoroughly  tested in humans. New generation recombinants retargeted to cancer-specific heterologous receptors have been generated and are presently evaluated in pre-clinical settings. They will be reviewed along with the molecular bases of cancer specificity and the strategies for the enhancement of oncolytic potential of HSV recombinants.
[Back to top]


Oncolytic Viruses for Induction of Anti-tumor Immunity
Alex W. Tong, Neil Senzer, Vincenzo Cerullo, Nancy Smyth Templeton, Akseli Hemminki, and John Nemunaitis
[Purchase Article] [PMID: 21740355 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00218]

Oncolytic virotherapy is an evolving but, as yet, unrealized treatment option for cancer.  This approach harnesses the cancer-restricted replicative activity of engineered viruses to achieve tumor cell kill.  Tumors that are resistant to chemotherapy or radiotherapy can be susceptible to viral oncolysis because of distinct cell kill mechanisms.  There is now compelling evidence that collateral induction of anti-tumor immune responses contributes substantially to viral anti-tumor activities. In addition to the expected anti-viral immune clearance, the "danger" signal created by virus-infected cells can generate immune co-stimulation known to override immune suppression and reverse tolerance within the tumor microenvironment.  Our recent findings indicate that immune activation augments the clinical outcomes of oncolytic virotherapy.  Strikingly similar and robust clinical response rates (>25%) were observed among advanced cancer patients following intratumoral treatments with adenoviral (AdD24) and herpes simplex (JS1/34.5-/47) constructs armed with an integrated granulocyte-macrophage colony-stimulating factor (GMCSF) payload.  Both agents produced regressions in injected as well as distant, uninjected lesions, demonstrating systemic effectiveness.  We discuss the innate and adaptive immune activating events that may contribute to these clinical outcomes, and examine systemic delivery strategies to tilt the immunological balance from viral clearance to tumor elimination.
[Back to top]


Bugs and Drugs: Oncolytic Virotherapy in Combination with Chemotherapy
Sonia Tusell Wennier, Jia Liu and Grant McFadden
[FULL-TEXT INQUIRY] [PMID: 21740354 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00219]

Single agent therapies are rarely successful in treating cancer, particularly at metastatic or end stages, and survival rates with monotherapies alone are generally poor. The combination of multiple therapies to treat cancer has already driven significant improvements in the standard of care treatments for many types of cancers.  The first combination treatments exploited for cancer therapy involved the use of several cytotoxic chemotherapy agents.  Later, with the development of more targeted agents, the use of novel, less toxic drugs, in combination with the more classic cytotoxic drugs has proven advantageous for certain cancer types.  Recently, the combination of oncolytic virotherapy with chemotherapy has shown that the use of these two therapies with very distinct anti-tumor mechanisms may also lead to synergistic interactions that ultimately result in increased therapeutic effects not achievable by either therapy alone.  The mechanisms of synergy between oncolytic viruses (OVs) and chemotherapeutic agents are just starting to be elucidated.  It is evident, however, that the success of these OV-drug combinations depends greatly on the particular OV, the drug(s) selected, and the cancer type targeted.  This review summarizes the different OV-drug combinations investigated to date, including the use of second generation armed OVs, which have been studied with the specific purpose of generating synergistic interactions with particular chemotherapy agents.  The known mechanisms of synergy between these OV-drug combinations are also summarized. The importance of further investigating these mechanisms of synergy will be critical in order to maximize the therapeutic efficacy of OV-drug combination therapies in the future.
[Back to top]


Fusogenic Oncolytic Herpes Simplex Viruses as a Potent and Personalized Cancer Vaccine
Qi-Xiang Li, Guohong Liu and Xiaoliu Zhang
[FULL-TEXT INQUIRY] [PMID: 21740353 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00220]

The recent FDA approval of Sipuleucel-T for the treatment of prostate cancer represents an important milestone of cancer immunotherapy, which, for the first time, validates the concept of bringing true clinical benefit to cancer patients by stimulating patients’ own anti-tumor immunity.  Among the different experimental cancer immunotherapies, oncolytic virotherapy may represent a low-cost yet potent and personalized cancer vaccine for the treatment of solid tumors.  This review describes the constructions of several human herpes simplex virus (HSV)-derived oncolytic viruses as candidate cancer vaccines, which induce specific and potent anti-tumor immunity in pre-clinical models, and thus resulting in stronger overall anti-tumor efficacy as compared to oncolytic effect alone.  This article also describes the approaches to enhance the antitumor immunity of oncolytic HSVs, and in particular, the key role played by integrating membrane-fusion activity into these viruses.  Additionally, this article reviews the potential effect of certain chemotherapeutic agents (e.g. cyclophosphamide) in boosting antitumor immunity induced by oncolytic HSV, and the mechanisms behind it.  In summary, all the preclinical and clinical data have suggested that HSV-based oncolytic virotherapies could likely be developed as a new generation of cancer vaccines for the treatment of solid tumors.
[Back to top]


In search of a consensus terminology in the field of platelet concentrates for surgical use: Platelet-Rich Plasma (PRP), Platelet-Rich Fibrin  (PRF), fibrin gel polymerization and leukocytes
David M. Dohan Ehrenfest, Tomasz Bielecki, Allan Mishra, Piero Borzini, Francesco Inchingolo, Gilberto Sammartino, Lars Rasmusson and Peter A. Everts
[FULL-TEXT INQUIRY] [PMID: 21740379 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00222]

In the field of platelet concentrates for surgical use, most products are termed Platelet-Rich Plasma (PRP). Unfortunately, this term is very general and incomplete, leading to many confusions in the scientific database. In this article, a panel of experts discusses this issue and proposes an accurate and simple terminology system for platelet concentrates for surgical use. Four main categories of products can be easily defined, depending on their leukocyte content and fibrin architecture: Pure Platelet-Rich Plasma (P-PRP), such as cell separator PRP, Vivostat PRF or Anitua’s PRGF; Leukocyte- and Platelet-Rich Plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan, Angel or GPS PRP; Pure Plaletet-Rich Fibrin (P-PRF), such as Fibrinet; and Leukocyte- and Platelet-Rich Fibrin (L-PRF), such as Choukroun’s PRF. P-PRP and L-PRP refer to the unactivated liquid form of these products, their activated versions being respectively named P-PRP gels and L-PRP gels. The purpose of this search for a terminology consensus is to plead for a more serious characterization of these products. Researchers have to be aware of the complex nature of these living biomaterials, in order to avoid misunderstandings and erroneous conclusions. Understanding the biomaterials or believing in the magic of growth factors ? From this choice depends the future of the field.
[Back to top]


Platelet concentrates for topical use: bedside device and blood transfusion technology. Quality and Versatility
Piero Borzini, Valeria Balbo, Laura Mazzucco
[FULL-TEXT INQUIRY] [PMID: 21740378 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00223]

More or less after a decade of experimental and pioneering manual procedures to prepare platelet-rich plasma (PRP) for topical use, several portable and bedside devices were made available to prepare the PRP at the point-of-care. This technical opportunity increased the number of patients who got access to the treatment with autologous PRP and PRP-gel. Since topical treatment of tissue with PRP and PRP-gel was restricted to autologous preparation, blood transfusion centers that professionally prepare donor-derived platelet concentrates were not able to cover the overwhelming request for autologous PRP supply. Principally for logistic and organization reasons blood transfusion centers usually fail the challenge of prompt delivery of PRP to the physician over large territory. Nevertheless the blood bank production of platelet concentrates is associated with high standardization and quality controls not achievable from bedside and portable devices. Furthermore it easy to demonstrate that high-volume blood bank-produced platelet concentrates are less expensive than low-volume PRP produced by portable and bedside devices. Taking also in consideration the ever-increasing safety of the blood components, the relationship between bedside device-produced and blood-bank-produced PRP might be reconsidered. Here we discuss this topic concluding that the variety of sources of PRP production is an opportunity for versatility and that, ultimately, versatility is an opportunity for the patient’s care.
[Back to top]


Do the fibrin architecture and leukocyte content influence the growth factor release of platelet concentrates ? An evidence-based answer comparing a Pure Platelet-Rich Plasma (P-PRP) Gel and a Leukocyte- and Platelet-Rich Fibrin (L-PRF).
David M. Dohan Ehrenfest, Tomasz Bielecki, Ryo Jimbo, Giovanni Barbé, Marco Del Corso, Francesco Inchingolo and Gilberto Sammartino
[FULL-TEXT INQUIRY] [PMID: 21740377 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00224]

Platelet concentrates for surgical use are tools of regenerative medicine designed for the local release of platelet growth factors into a surgical or wounded site, in order to stimulate tissue healing or regeneration. Leukocyte content and fibrin architecture are 2 key characteristics of all platelet concentrates and allow to classify these technologies in 4 families, but very little is known about the impact of these 2 parameters on the intrinsic biology of these products. In this demonstration, we highlight some outstanding differences in the growth factor and matrix protein release between 2 families of platelet concentrate: Pure Platelet-Rich Plasma (P-PRP, here the Anitua’s PRGF - Preparation Rich in Growth Factors - technique) and Leukocyte- and Platelet-Rich Fibrin (L-PRF, here the Choukroun’s method). These 2 families are the extreme opposites in terms of fibrin architecture and leukocyte content. The slow release of 3 key growth factors (Transforming Growth Factor β1 (TGFβ1), Platelet-Derived Growth Factor AB (PDGF-AB) and Vascular Endothelial Growth Factor (VEGF)) and matrix proteins (fibronectin, vitronectin and thrombospondin-1) from the L-PRF and P-PRP gel membranes in culture medium is described and discussed. During 7 days, the L-PRF membranes slowly release significantly larger amounts of all these molecules than the P-PRP gel membranes, and the 2 products display different release patterns. In both platelet concentrates, vitronectin is the sole molecule to be released almost completely after only 4 hours, suggesting that this molecule is not trapped in the fibrin matrix and not produced by the leukocytes. Moreover the P-PRP gel membranes completely dissolve in the culture medium after less than 5 days only, while the L-PRF membranes are still intact after 7 days. This simple demonstration shows that the polymerization and final architecture of the fibrin matrix considerably influence the strength and the growth factor trapping/release potential of the membrane. It also suggests that the leukocyte populations have a strong influence on the release of some growth factors, particularly TGFβ1. Finally, the various platelet concentrates present very different biological characteristics, and an accurate definition and characterization of the different families of product is a key issue for a better understanding and comparison of the reported clinical effects of these surgical adjuvants.
[Back to top]


The role of leukocytes from L-PRP/L-PRF in wound healing and immune defense: new perspectives
Tomasz Bielecki,  David M. Dohan Ehrenfest, Peter A. Everts and Andrzej Wiczkowski
[FULL-TEXT INQUIRY] [PMID: 21740376 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00225]

Platelet concentrates for topical use are innovative tools of regenerative medicine and their effects in various therapeutical situations are hotly debated. Unfortunately, this field of research mainly focused on the platelet growth factors, and the fibrin architecture and the leukocyte content of these products are too often neglected. In the four families of platelet concentrates, 2 families contain significant concentrations of leukocytes: L-PRP (Leukocyte- and Platelet-Rich Plasma) and L-PRF (Leukocyte- and Platelet-Rich Fibrin). The presence of leukocytes has a great impact on the biology of these products, not only because of their immune and antibacterial properties, but also because they are turntables of the wound healing process and the local factor regulation. In this article, the various kinds of leukocytes present in a platelet concentrate are described (particularly the various populations of granulocytes and lymphocytes), and we insist on the large diversity of factors and pathways that these cells can use to defend the wound site against infections and to regulate the healing process. Finally, the impact of these cells in the healing properties of the L-PRP and L-PRF is also discussed: if antimicrobial properties were already pointed out, effects in the regulation of cell proliferation and differentiation were also hypothesized. Leukocytes are key actors of many platelet concentrates, and a better understanding of their effects is an important issue for the development of these technologies.
[Back to top]


Is the use of autologous platelet-rich plasma gels in gynecologic, cardiac, and general, reconstructive surgery beneficial?
Peter A.M. Everts, Maarten M. Hoogbergen, Tjaarda A.Weber, Roger J.J. Devilee, Gust van Monftort, Ignace H.J.T. de Hingh
[FULL-TEXT INQUIRY] [PMID: 21740375 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00226]

Tissue repair at wound sites begins with clot formation, and subsequently platelet degranulation with the release of platelet growth factors, which are necessary and well-regulated processes to achieve wound healing. Platelet-derived growth factors are biologically active substances that enhance tissue repair mechanisms, such as chemotaxis, cell proliferation, angiogenesis, extracellular matrix deposition, and remodeling. This review describes the biological background and results on the topical use of autologous platelet-rich plasma and platelet gel in gynecologic, cardiac, and general surgical procedures, including chronic wound management and soft-tissue injuries.
[Back to top]


Applications of leukocyte- and platelet-rich plasma (L-PRP) in trauma surgery
Ting Yuan, Shang-Chun Guo, Pei Han, Chang-Qing Zhang, Bing-Fang Zeng
[FULL-TEXT INQUIRY] [PMID: 21740374 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00227]

Leukocyte- and platelet-rich plasma (L-PRP) contains high concentrations of platelet, leukocytes and other bioactivities, which play an prominent role in both bone and soft tissue healing processes. Large numbers of studies provide evidence for application of L-PRP in experiments and clinical practice. It has been identified to improve cellular chemotaxis, proliferation and differentiation, angiogenesis, and production of extracellular matrix, but also responsible for stimulating defense mechanisms against infections. L-PRP is now playing an increasing role in the management of patients with traumatic injuries. However, most studies are only anecdotal or case reports, and then larger controlled studies are needed. This article introduces the reader to L-PRP properties and L-PRP applications in trauma surgery, including applications of L-PRP in bone healing, acute soft tissue wound healing, and repairing of acute muscle, tendon, ligament, nerve and cartilage injury caused by trauma.
[Back to top]


Sports Medicine Applications of Platelet Rich Plasma
Allan Mishra, Kimberly Harmon, James Woodall, and Amy Vieira
[FULL-TEXT INQUIRY] [PMID: 21740373 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00228]

Platelet rich plasma (PRP) is a powerful new biologic tool in sports medicine. PRP is a fraction of autologous whole blood containing and increased number of platelets and a wide variety of cytokines such as platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF) and transforming growth factor beta-1 (TGF-B1), fibroblast growth factor (FGF), Insulin-like growth factor-1 (IGF-1) among many others.  Worldwide interest in this biologic technology has recently risen sharply.  Basic science and preclinical data support the use of PRP for a variety of sports related injuries and disorders.   The published, peer reviewed, human data on PRP is limited.    Although the scientific evaluation of clinical efficacy is in the early stages, elite and recreational athletes already use PRP in the treatment of sports related injuries.  Many questions remain to be answered regarding the use of PRP including optimal formulation, including of leukocytes, dosage and rehabilitation protocols.  In this review, a classification for platelet rich plasma is proposed and the in-vitro, preclinical and human investigations of PRP applications in sports medicine will be reviewed as well as a discussion of rehabilitation after a PRP procedure. The regulation of PRP by the World Anti-Doping Agency will also be discussed. PRP is a promising technology in sports medicine; however, it will require more vigorous study in order to better understand how to apply it most effectively.
[Back to top]


Leukocyte- and platelet-rich fibrin (L-PRF) for long-term delivery of growth factor in rotator cuff repair: review, preliminary results and future directions
Matthias A. Zumstein, Simon Berger Martin Schober, Pascal Boileau, Richard W. Nyffeler, Michael Horn, and Clemens A. Dahinden
[FULL-TEXT INQUIRY] [PMID: 21740372 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00229]

Surgical repair of the rotator cuff repair is one of the most common procedures in orthopedic surgery. Despite it being the focus of much research, the physiological tendon-bone insertion is not recreated following repair and there is an anatomic non-healing rate of up to 94%. During the healing phase, several growth factors are upregulated that induce cellular proliferation and matrix deposition. Subsequently, this provisional matrix is replaced by the definitive matrix. Leukocyte- and platelet-rich fibrin (L-PRF) contain growth factors and has a stable dense fibrin matrix. Therefore, use of L-PRF in rotator cuff repair is theoretically attractive. The aim of the present study was to determine 1) the optimal protocol to achieve the highest leukocyte content; 2) whether L-PRF releases growth factors in a sustained manner over 28 days; 3) whether standard/gelatinous or dry/compressed matrix preparation methods result in higher growth factor concentrations. 1) The standard L-PRF centrifugation protocol with 400 x g showed the highest concentration of platelets and leukocytes. 2) The L-PRF clots cultured in medium showed a continuous slow release with an increase in the absolute release of growth factors TGF-β1, VEGF and MPO in the first 7 days, and for IGF1, PDGF-AB and platelet activity (PF4=CXCL4) in the first 8 hours, followed by a decrease to close to zero at 28 days. Significantly higher levels of growth factor were expressed relative to the control values of normal blood at each culture time point. 3) Except for MPO and the TGFβ-1, there was always a tendency towards higher release of growth factors (ie, CXCL4, IGF-1, PDGF-AB, and VEGF) in the standard/gelatinous- compared to the dry/compressed group. L-PRF in its optimal standard/gelatinous-type matrix can store and deliver locally specific healing growth factors for up to 28 days and may be a useful adjunct in rotator cuff repair.
[Back to top]


Current knowledge and perspectives for the use of Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF) in oral and maxillofacial surgery. Part 1: periodontal and dentoalveolar surgery
Marco Del Corso, Alain Vervelle, Alain Simonpieri, Ryo Jimbo, rancesco Inchingolo, Gilberto Sammartino and David M. Dohan Ehrenfest
[FULL-TEXT INQUIRY] [PMID: 21740371 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00230]

Platelet concentrates for surgical use are innovative tools of regenerative medicine, and were widely tested in oral and maxillofacial surgery. Unfortunately, the literature on the topic is contradictory and the published data are difficult to sort and interpret. In periodontology and dentoalveolar surgery, the literature is particularly dense about the use of the various forms of Platelet-Rich Plasma (PRP) - Pure Platelet-Rich Plasma (P-PRP) or Leukocyte- and Platelet-Rich Plasma (L-PRP) - but still limited about Platelet-Rich Fibrin (PRF) subfamilies. In this first article, we describe and discuss the current published knowledge about the use of PRP and PRF during tooth avulsion or extraction, mucogingival surgery, Guided Tissue Regeneration (GTR) or bone filling of periodontal intrabony defects, and regeneration of alveolar ridges using Guided Bone Regeneration (GBR), in a comprehensive way and in order to avoid the traps of a confusing literature and to highlight the underlying universal mechanisms of these products. Finally, we particularly insist on the perspectives in this field, through the description and illustration of the systematic use of L-PRF (Leukocyte- and Platelet-Rich Fibrin) clots and membranes during tooth avulsion, cyst exeresis or the treatment of gingival recessions by root coverage. The use of L-PRF also allowed to define new therapeutic principles: NTR (Natural Tissue Regeneration) for the treatment of periodontal intrabony lesions and Natural Bone Regeneration (NBR) for the reconstruction of the alveolar ridges. In periodontology, this field of research will soon find his golden age by the development of user-friendly platelet concentrate procedures, and the definition of new efficient concepts and clinical protocols.
[Back to top]


Current knowledge and perspectives for the use of Platelet-Rich Plasma (PRP) and Platelet-Rich Fibrin (PRF) in oral and maxillofacial surgery. Part 2: bone graft, implant and reconstructive surgery
Alain Simonpieri, Marco Del Corso, Alain Vervelle, Ryo Jimbo, Francesco Inchingolo, Gilberto Sammartino and David M. Dohan Ehrenfest
[FULL-TEXT INQUIRY] [PMID: 21740370 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00231]

Platelet concentrates for surgical use are innovative tools of regenerative medicine, and were widely tested in oral and maxillofacial surgery. Unfortunately, the literature on the topic is contradictory and the published data are difficult to sort and interpret. In bone graft, implant and reconstructive surgery, the literature is particularly dense about the use of the various forms of Platelet-Rich Plasma (PRP) - Pure Platelet-Rich Plasma (P-PRP) or Leukocyte- and Platelet-Rich Plasma (L-PRP) - but still limited about Platelet-Rich Fibrin (PRF) subfamilies. In this second article, we describe and discuss the current published knowledge about the use of PRP and PRF during implant placement (particularly as surface treatment for the stimulation of osseointegration), the treatment of peri-implant bone defects (after peri-implantitis, during implantation in an insufficient bone volume or during immediate post-extraction or post-avulsion implantation), the sinus-lift procedures and various complex implant-supported treatments. Other potential applications of the platelet concentrates are also highlighted in maxillofacial reconstructive surgery, for the treatment of patients using bisphosphonates, anticoagulants or with post-tumoral irradiated maxilla. Finally, we particularly insist on the perspectives in this field, through the description and illustration of the use of L-PRF (Leukocyte- and Platelet-Rich Fibrin) clots and membranes during the regeneration of peri-implant bone defects, during the sinus-lift procedure and during complex implant-supported rehabilitations. The use of L-PRF allowed to define a new therapeutic concept called the Natural Bone Regeneration (NBR) for the reconstruction of the alveolar ridges at the gingival and bone levels. As it is illustrated in this article, the NBR principles allow to push away some technical limits of global implant-supported rehabilitations, particularly when combined with other powerful biotechnological tools: metronidazole solution, adequate bone substitutes and improved implant designs and surfaces (for example here AstraTech Osseospeed or Intra-Lock Ossean implants). As a general conclusion, we are currently living a transition period in the use of PRP and PRF in oral and maxillofacial surgery. PRPs failed to prove strong strategic advantages that could justify their use in daily practice, and the use of most PRP techniques will probably be limited to some very specific applications where satisfactory results have been reached. Only a few simple, inexpensive and efficient techniques such as the L-PRF will continue to develop in oral and maxillofacial surgery in the next years. This natural evolution illustrates that clinical sciences need concrete and practical solutions, and not hypothetical benefits. The history of platelet concentrates in oral and maxillofacial surgery finally demonstrates also how the techniques evolve and sometimes promote the definition of new therapeutical concepts and clinical protocols in the today’s era of regenerative medicine.
[Back to top]


The Role of “Eye Platelet Rich Plasma” (E-Prp) for  Wound Healing in Ophthalmology
Jorge L. Alió, Francisco Arnalich-Montiel, Alejandra E. Rodriguez
[Purchase Article] [PMID: 21740369 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00232]

Blood derived products have demonstrated their capacity to enhance healing and stimulate the regeneration of different tissues and this enhancing effect is attributed to the growth factors and bioactive proteins that are synthesized and present in blood. Eye platelet rich plasma (E-PRP) provides higher concentration of essential growth factors and cell adhesion molecules by concentrating platelets in a small volume of plasma as compared with autologous serum, the latter being used widely in ophthalmology for epithelial wound healing of the cornea for the last two decades. These growth factors and cell adhesion molecules have a major role in wound healing and enhance the physiological process at the site of the injury/surgery via eye drops or clot. E-PRP has been used more recently, and has achieved successful outcomes in peer-review articles in the treatment of dormant ulcers (epithelial defects of the cornea that fail to heal), moderate to severe dry eye syndrome, ocular surface syndrome post Laser In Situ Keratomileusis (LASIK), and for surface reconstruction after corneal perforation associated with amniotic membrane transplantation. Preparation of E-PRP in the two available formulations, eyedrops and clot, is inexpensive and easy although it requires following strict sterility conditions using sterile and disposable materials and operating inside a laminar flow hood. No serious adverse effects have been described with the use of these products, and it is generally well tolerated. In summary, Platelet enriched plasma in the form obtained in ophthalmology, E-PRP, is a reliable and effective therapeutic tool to enhance epithelial wound healing in ocular surface disease.
[Back to top]


L-PRP/L-PRF in esthetic plastic surgery, regenerative medicine of the skin and chronic wounds
Agata Cieslik-Bielecka, Joseph Choukroun, Guillaume Odin and David M. Dohan Ehrenfest
[FULL-TEXT INQUIRY] [PMID: 21740368 PubMed - indexed for MEDLINE] [BSP/CPB/E-Pub/00233]

The use of platelet concentrates for topical use is of particular interest for the promotion of skin wound healing. Fibrin-based surgical adjuvants are indeed widely used in plastic surgery since many years in order to improve scar healing and wound closure. However, the addition of platelets and their associated growth factors opened a new range of possibilities, particularly for the treatment of chronic skin ulcers and other applications of regenerative medicine on the covering tissues. In the 4 families of platelet concentrates available, 2 families were particularly used and tested in this clinical field: L-PRP (Leukocyte- and Platelet-rich Plasma) and L-PRF (Leukocyte- and Platelet-Rich Fibrin). These 2 families have in common the presence of significant concentrations of leukocytes, and these cells are important in the local cleaning and immune regulation of the wound healing process. The main difference between them is the fibrin architecture, and this parameter considerably influences the healing potential and the therapeutical protocol associated to each platelet concentrate technology. In this article, we describe the historical evolutions of these techniques from the fibrin glues to the current L-PRP and L-PRF, and discuss the important functions of the platelet growth factors, the leukocyte content and the fibrin architecture in order to optimize the numerous potential applications of these products in regenerative medicine of the skin. Many outstanding perspectives are appearing in this field and require further research.
[Back to top]


Ectopic Lymphoid Neogenesis And Lymphoid Chemokines In Sjogren’s Syndrome: At The Interplay Between Chronic Inflammation, Autoimmunity And Lymphomagenesis
Michele Bombardieri and Costantino Pitzalis
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00248]

It has long been demonstrated that a subset of patients with Sjogren’s syndrome (SS) develop ectopic lymphoid structures (ELS) in the salivary glands (SG). These structures are characterised by periductal clusters of T and B lymphocytes, development of high endothelial venules and differentiation of follicular dendritic cells (FDC) networks. Evidence in patients with and animal models of SS demonstrated that the formation and maintenance of ELS in the SG is critically dependent on the ectopic expression of lymphotoxins (LT) and lymphoid chemokines CXCL13, CCL19, CCL21 and CXCL12. Several cell types, including resident epithelial, stromal and endothelial cells as well as different subsets of infiltrating immune cells, have been shown to be capable of producing some of these factors during chronic inflammation in SS. In this review we focus on the cellular and molecular mechanisms regulating the formation of ELS in SS SG, with particular emphasis on the role of lymphoid chemokines. In addition, we summarise accumulating data in support of the notion that ELS in SS represent functional niches whereby autoreactive B cells undergo affinity maturation, clonal selection and differentiation into autoantibody producing cells, thus contributing to autoimmunity over and above secondary lymphoid organs. Furthermore, we review the emerging role of ELS and lymphoid chemokines in driving extranodal B cell lymphomagenesis in SS and we focus on recent evidence suggesting that ELS identify subsets of SS patients at increased risk of developing systemic manifestations and lymphoma.
[Back to top]


Autoantibodies and Sjögren’s syndrome:a physiologist’s perspective
Peter M. Smith and Luke J Dawson
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00249]

Sjögren’s syndrome is a systemic autoimmune condition centred around salivary gland dysfunction and atrophy. There are a plethora of antibodies that mark the decline of the salivary glands, most of which relate to apoptopic mediated destruction of acinar cells. The best known of these autoantibodies, anti-Ro and anti-La form part of the diagnostic criteria for the condition. An emerging viewpoint in recent years is that glandular dysfunction precedes rather than follows glandular atrophy and attention has shifted to the interface between the immune system and the secretory process. An autoantibody against the muscarinic type 3 acetylcholine receptor occupies precisely this position because it inhibits the acetylcholine receptor which controls salivary and lacrimal fluid secretion. The consequence of identifying an autoantibody that can cause rather than simply reflect the disease process are manifold and could have a huge impact on development of therapeutic treatments. Furthermore, a “functional” autoantibody directed against an important component of the autonomic nervous system could also account for some of the extraglandular features of Sjögren’s syndrome.
[Back to top]


B-Lymphocytes Govern The Pathogenesis Of Sjögren's Syndrome
Pierre Youinou, Alain Saraux and Jacques-Olivier Pers
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00250]

T cells have originally occupied central stage of the debate on the type of lymphocytes governing the pathogenesis of Sjögren’s syndrome (SS). However, B cells has since been substituted for T cells, and insights into their functions have revealed that they accomplish various tasks. Beyond the paradigm that T lymphocytes maintain strict control over B lymphocytes, these latter cells solicit their own help from the former, release a flurry of cytokines, and act as antigen-presenting cells. In SS, excessive of the B cell-activating factor (BAFF) may cause B-cell quantitative anomalies, such as inflation of mature B (Bm)2/Bm2’ cells in the circulation, or accumulation of transitional type 2, marginal zone (MZ) and memory B cells within the exocrine gland infiltrates. These excesses are also associated with B-cell qualitative anomalies, such as the internal synthesis of BAFF, and a default mechanism that promotes the autoantibody production in ectopic germinal centers or MZ equivalents. Thus, SS should rather be conceived as a quintessential model for B cell-induced autoimmunity. Such a view opens novel prospects for treatment, and indeed B cell-ablative therapy has already been shown to be beneficial to these patients.
[Back to top]


Biologic Therapies in Primary Sjögren's syndrome
Dr S J Bowman
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00251]

Primary Sjögren’s Syndrome (PSS) is characterized by dryness of the eyes and mouth due to lymphocytic infiltration of secretory exocrine glands. As well as disabling dryness, patients commonly have fatigue and arthralgia and an associated reduction in quality of life. The condition principally affects adult women and is relatively common – approximately 1:1000 to 1:250 adult women are estimated to have the condition in European/North American studies.

Current therapy is principally symptomatic with the use of artificial tears and oral gels, pastilles and sprays. Medications to stimulate residual glandular secretion can be helpful for appropriate individuals. A proportion of patients also develop extraglandular features such as skin vasculitis, or lung, neurological, haematological or other systemic involvement. Conventional general immunosuppressive therapies such as corticosteroids or disease-modifying drugs, have been used in some patients with these clinical features.

Biologic therapies specifically directed against molecules involved in disease pathogenesis represent a potentially more effective approach to therapeutic intervention in rheumatic diseases including PSS. The greatest experience in PSS is with rituximab, an anti-B-cell monoclonal antibody already in use for the treatment of B-cell lymphoma and rheumatoid arthritis. A randomised placebo controlled study is currently recruiting in France and a further study is planned in the UK.

This review discusses the utility of biologic therapies in PSS, potential challenges for their use, the available data on rituximab and the potential role for other biologic therapies currently in development, or in clinical trials, in other autoimmune conditions.
[Back to top]


Primary Sjögren’s Syndrome : Time For Prospective Cohorts
Jacques-Eric Gottenberg and Xavier Mariette
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00252]

Many important clinical questions concerning primary Sjögren’s syndrome (pSS) remains, that can only be adequately addressed by prospective cohorts.

Thus, a better knowledge of the clinical outcome, the course of disease activity and the risk factors of lymphoma requires the setting up of cohorts with biobanks. The homogeneous collection of clinical data, disease activity, patient-related outcome, and biological samples, including DNA, RNA and serum, is definitively mandatory to determine new biological prognostic factors and identify disease activity markers. Three large prospective cohorts have already started to enroll patients with pSS. This will be highly invaluable for scientists and clinicians to gain a better insight into the pathogenesis of pSS, as well as to identify  prognostic markers and new therapeutic targets.
[Back to top]


Current And Future Challenges In Primary Sjögren’s Syndrome
Arjan Vissink, Hendrika Bootsma, Fred KL Spijkervet, Shen Hu, David T Wong and  Cees GM Kallenberg
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00253]

Sjögren’s syndrome (SS) is an autoimmune inflammatory disorder of exocrine glands. SS particularly affects the lacrimal and salivary glands. Dry mouth and dry eyes are frequently proffered as presenting symptoms, but non-specific symptoms such as malaise and fatigue, and extraglandular manifestations like purpura, polyneuropathy and arthritis are also often present. Moreover, lymphomas develop in about 7.5% of SS patients, mostly marginal zone B-cell lymhomas. Futhermore, SS has a very substantial impact on the patients’ quality of life and their daily activities.

Recently, many breakthroughs have been seen in salivary diagnostics, which not only can be used for diagnosis but also for monitoring of disease activity and disease progression as well as for objectively scoring the effect of intervention treatment with biologicals. In addition, salivary proteomics, genomics and system biology have been shown to be very promising tools in unravelling the pathophysiology of SS, thus providing in depth insight in its underlying mechanisms which could give clues for intervention therapies with biologicals. The latter is of particular interest as B cell depletion therapy has been shown a very promising therapy for a subgroup of SS patients. When applying salivary diagnostics in combination with instruments to rate disease activity and progression in SS, one might be able to select those SS patients who respond to a particular type of biological. These topics are addressed in this review and promises for the near future are described.
[Back to top]


Primary Sjögren’s Syndrome And The Type I Interferon System
Gunnel Nordmark, Maija-Leena Eloranta and  Lars Rönnblom
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00254]

Patients with primary Sjögren’s syndrome (pSS) have an activated type I interferon (IFN) system that contribute to the etiopathogenesis and clinical manifestations of the disease. The type I IFN system consists of the stimuli for type I IFN production, the receptors, cells and transcription factors involved in the synthesis of type I IFNs, the type I IFN-receptor and the effects on target cells. Increased type I IFN activity has been demonstrated in sera from patients with pSS and IFN-α, the main type I IFN, has been detected in the minor salivary glands. Gene expression profiling of peripheral blood mononuclear cells (PBMCs) and minor salivary glands from pSS patients display an up-regulation of type I IFN-induced genes, an “IFN signature”. The professional IFN-α producing plasmacytoid dendritic cell (pDC) shows a reduced frequency in the peripheral blood, but has been detected in the salivary glands, possibly due to tissue recruitment. Polymorphisms in the interferon regulatory factor 5 (IRF5) and signal transducer and activator of transcription 4 (STAT4) genes in the type I IFN system, are associated with increased risk for pSS. A postulated disease model is that an initial viral infection induces type I IFN production in the salivary glands with subsequent activation of the adaptive immune system resulting in the production of autoantibodies against the RNA-binding proteins SSA/SSB/RNP. Interferogenic immune complexes are formed, which trigger the pDCs to an ongoing type I IFN production, which sustain the disease process. Potential therapeutic targets can be identified within the type I IFN system.
[Back to top]


Local Activation and Systemic Dysregulation of T Lymphocytes in Sjögren’s Syndrome
Robert Busch, Andreas V. Hadjinicolaou and Frances C. Hall
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00255]

T cells are implicated in both local and systemic pathophysiology of primary Sjögren’s syndrome (PSS). Lymphocytic infiltrates in exocrine glands are dominated by CD4+ T cells, some contributing to ectopic lymphoid tissue, others, unusually, exhibiting cytotoxic potential. Cytokine secretion patterns are complex, with Th1 and Th17 components implicated in pathology. Circulating T cells exhibit phenotypes consistent with hyperactivation, cytokine imbalance, and homeostatic alterations; CD4 lymphopenia is recognized as a risk factor for developing lymphoma. Evidence of oligoclonal expansion is found locally and systemically. Functional alterations (e.g. cytokine secretion profile, migratory potential, target cell interactions) are less clearly defined. Attempts at T cell-targeted therapy of PSS have been limited, although therapy targeted at other arms of the immune response may also affect T cells. A better understanding of T-cell dysregulation in PSS is required in order to understand its contribution to disease, aid prognosis, and improve therapeutic interventions aimed at this aspect of the disease.
[Back to top]


Epigenetics and Sjögren’s syndrome
Christelle Le Dantec, Marie-Michèle Varin, Wesley H Brooks, Jacques-Olivier Pers, Pierre Youinou and Yves Renaudineau
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00256]

There is growing evidence that epigenetics, the study of heritable changes in gene expression that do not involve mutations in the DNA itself, may play an essential role in autoimmune diseases (AID). In Sjögren’s syndrome (SS), a chronic AID characterized by an epithelis of the exocrine glands, epigenetic studies have focused on three mechanisms: DNA methylation and its consequences including human endogenous retrovirus (HERV) expression; microRNA expression; and protein post-translational modifications associated with autoantibody production. Although in its infancy, comprehension of the epigenetic (dys)regulation in SS may help us to understand: why SS affects predominantly middle-aged women; why genetically predisposed individuals develop SS but not others; why flare-ups occur; why treatment responses differ between patients; and why some patients develop lymphoma. From these studies will arise a better comprehension of the pathophysiology of SS as well as development of new diagnostic and prognostic biomarkers, and novel therapeutics for prevention and perhaps early intervention.
[Back to top]


Cell Hierarchy, Metabolic Flexibity And Systems Approaches To Cancer Treatment
Patries M. Herst and Michael V. Berridge
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00258]

The proliferative cancer cell paradigm that has driven cancer drug development for the past 50 years has failed to generate treatments that cure most metastatic adult cancers.  This view is supported not only by cumulative experience with conventional cytotoxic anticancer drugs, but also by the application of highly-targeted anticancer compounds against, for example, BCR-ABL in CML and mutant BRAF in metastatic melanoma.  Such drugs often send their respective cancers into complete molecular remission but fail to effect cures because a small population of quiescent or slowly self-renewing cancer cells that are drug and radiation resistant survive treatment indefinitely.  This review explores the grounds for an emerging cancer paradigm that views cancer as a disorganized tissue with hierarchical cellular compartments in which the boudaries are less well-defined than in normal tissues with plasticity controlled by epigenetic changes mediated by the local microenvironment.  Increased metabolic flexibility and adaptability give cancer cells an additional survival advantage that may be able to be targeted with drugs like metformin. Combining approaches that target the increased metabolic flexibility of cancer cells as well as ablating rapidly-proliferating cells and self-renewing cancer stem cells in individual cancers are needed to address the holy grail of cancer cure.
[Back to top]


p53 Regulation of Energy Metabolism and Mitochondria Regulation of p53 in Cancer Cells: An Insight into the Role of Manganese Superoxide Dismutase
Yulan Sun, Aaron K. Holley and Daret K. St. Clair
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00259]

Accumulated evidence suggests that p53 plays an important role in the regulation of metabolism and intracellular redox homeostasis through transcription-dependent and -independent mechanisms. Mitochondria, the power plant of cells, provide cells with ATP for their functions by regulating energy metabolism. In addition, as the byproducts of metabolism, reactive oxygen species (ROS) generated in the mitochondria can serve as signaling molecules to regulate p53 function. The regulation of p53 by mitochondria, especially redox-mediated regulation, may be involved in controlling the cellular switch between survival and death. The interplay between p53 and manganese superoxide dismutase (MnSOD), an important mitochondrial antioxidant enzyme, is an example of how nuclear and mitochondrial p53 coordinate their response to different levels of stress and contribute to the fate of cells.
[Back to top]


Combined Chemotherapy or Biotherapy with Jasmonates: Targeting Energy Metabolism for Cancer Treatment
Uri Elia and Eliezer Flescher
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00260]

Mitochondria are known to play a key role in various cellular processes essential to both the life and death of cells, including calcium homeostasis, programmed cell death, and energy metabolism. Over 80 years ago, Otto Warburg discovered that in contrast to normal cells which produce most of their ATP via mitochondrial oxidative phosphorylation, cancer cells preferentially utilize glycolysis for production of ATP, a phenomenon known today as the "Warburg effect", and one which has been of great importance in the emergence of novel drugs and chemotherapeutic agents specifically targeting cancer cells. Several groups have reported in recent years that members of the plant stress hormones family of jasmonates, and some of their synthetic derivatives, exhibit anti-cancer activity in vitro and in vivo. Jasmonates have been shown to act directly on mitochondria of cancer cells, leading to mitochondrial swelling, membrane depolarization and cytochrome c release. Throughout the last few years, different groups have demonstrated that combination of jasmonates and various cytotoxic and chemotherapeutic agents yielded a synergistic cytotoxic effect. These results have been demonstrated in a variety of different cancer cell lines and may provide a strong basis for future clinical treatments which involve combination of MJ and different anti-cancerous agents. The potential synergistic effect may allow reduction of the administered dose, decrease of unwanted side effects, and reduction of the likelihood that the tumor will display resistance to the combined therapy.
[Back to top]


Regulation of the metabolism of polyunsaturated fatty acids and butyrate in colon cancer cells
Jiřina Hofmanová, Alena Vaculová and Alois Kozubík
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00261]

Experimental and epidemiological evidence supports the idea that dietary fat and fiber influence colon carcinogenesis. Particularly, their components, n-3 polyunsaturated fatty acids (PUFAs) and butyrate, have been proven to exhibit beneficial effects on colon epithelial cell metabolism, signaling, and kinetics, thus preventing colon inflammation and cancer. Moreover, these effects may be strengthened by PUFA and butyrate combination. It appears that administration of these compounds might be a relatively nontoxic form of supportive therapy improving cancer treatment outcomes and slowing down or preventing recurrence of certain types of cancer. However, their efficient application has to be based on solid scientific evidence of their mechanisms of action from the molecular and cellular to the organismal level. In this review, we emphasize the role of lipids and their metabolism during tumor development, describe some important mechanisms considering cellular and molecular levels of PUFA and butyrate action in colon epithelial cells, and particularly focus on the interaction of their metabolism and the signaling pathways with respect to the differences in response of normal and cancer colon cells.
[Back to top]


Use of anti-cancer drugs, mitocans, to enhance the immune responses against tumors
Hahn, T. Polanczyk, M.J. Borodovsky, A. Ramanathapuram and L.V. Akporiaye E.T Ralph, S.J.
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00262]

Cytotoxic drugs in cancer therapy are used with the expectation of selectively killing and thereby eliminating the offending cancer cells. If they should die in an appropriate manner, the cells can also release danger signals that promote an immune reaction that reinforces the response against the cancer. The identity of these immune-enhancing danger signals, how they work extra- and intracellularly, and the molecular mechanisms by which some anti-cancer drugs induce cell death to bring about the release of danger signals are the major focus of this review. A specific group of mitocans, the vitamin E analogs that act by targeting mitochondria to drive ROS production and also promote a more immunogenic means of cancer cell death exemplify such anti-cancer drugs. The role of reactive oxygen species (ROS) production and the events leading to the activation of the inflammasome and pro-inflammatory mediators induced by dying cancer cell mitochondria are discussed along with the evidence for their contribution to promoting immune responses against cancer.  Current knowledge of how the danger signals interact with immune cells to boost the anti-tumor response is also evaluated.
[Back to top]


Targeting the Mitochondrial Electron Transport Chain Complexes for the Induction of Apoptosis and Cancer Treatment
Jakub Rohlena, Lan-feng Dong and Jiri Neuzil
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00263]

Treatment of cancer is by no means universally successful and often manifests harmful side effects. The best way to improve the success rate and reduce the side effects would be to develop compounds that are able to kill cancer cells while leaving normal cells unaffected. In this respect, mitocans (an acronym from ‘mitochondria’ and ‘cancer’), a summary term we proposed for compounds that induce cell death by targeting mitochondria, show an encouraging trend. Here we provide an overview of mitocans specific for the mitochondrial electron transport chain. These mitocans are particularly interesting, because a frequent consequence of electron transport chain inhibition is the induction of superoxide formation resulting in the preferential killing of cancer cells, as these tend to be more sensitive than normal cells to sudden increases in oxidative stress. Furthermore, macromolecular complexes of the electron transport chain only rarely mutate in cancer, and represent useful targets for anti-cancer drug development when widely-applicable agents are sought.
[Back to top]


The warburg hypothesis and the atp supply in cancer cells Is oxidative phosphorylation impaired in malignant neoplasias?
Sara Rodríguez-Enríquez, Juan Carlos Gallardo-Pérez, Alvaro Marín-Hernández and Rafael Moreno-Sánchez
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00264]

Since Warburg proposed that cancer cells exhibit increased glycolysis due to apparent mitochondrial damage, numerous researchers have assumed that glycolysis is the predominant ATP supplier for cancer cell energy-dependent processes.  However, chemotherapeutic strategies using glycolytic inhibitors have been unsuccessful in arresting tumor proliferation indicating that the original Warburg proposal may not be applicable to all existing neoplasias.  This review analyzes recent information on mitochondrial metabolism in several malignant neoplasias emphasizing that, although tumor cells maintain a high glycolytic rate, the principal ATP production may derive from active oxidative phosphorylation.  Thus, anti-mitochondrial drug therapy may be an adequate strategy to arrest proliferation of oxidative phosphorylation-dependent neoplasias.
[Back to top]


Potentiation of Anti-Cancer Treatment by Modulators of Energy Metabolism
Maria C. Shoshan
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00265]

Oncogene-driven proliferative signaling in tumor cells requires comprehensive upregulation of cellular energy metabolism and macromolecule syntheses. These alterations are now known to include not only upregulated glycolysis, but also increased fatty acid metabolism, glutaminolysis, deregulated mitochondrial function and more. Many prospective targets for tumor-specific pharmacological modulation of metabolism have therefore been identified. While the prospective drugs do not necessarily show very high antitumor activity by themselves, they may by depriving tumor cells of energy and building blocks for repair and proliferation come to be of major clinical use as potentiators of standard chemotherapeutic drugs and/or radiation. To this end, not only inhibitors of specific enzyme functions are being investigated, but also drugs affecting the complex signaling networks that regulate organismal and cellular energy status. This review provides examples of how modulators of energy metabolism (MEMs) targetting different aspects of tumor cell metabolism have been found to potentiate cancer treatment in vitro and in vivo.
[Back to top]


The Potential of 11C-acetate PET for Monitoring the Fatty Acid Synthesis Pathway in Tumors
Laura M. DeFord-Watts, Akiva Mintz, and Steven J. Kridel
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00266]

Positron emission tomography (PET) is a molecular imaging modality that provides the opportunity to rapidly and non-invasively visualize tumors derived from multiple organs.  In order to do so, PET utilizes radiotracers, such as ¹⁸F-FDG and ¹¹C-acetate, whose uptake coincides with altered metabolic pathways within tumors.  Increased expression and activity of enzymes in the fatty acid synthesis pathway is a frequent hallmark of cancer cells.  As a result, this pathway has become a prime target for therapeutic intervention.  Although multiple drugs have been developed that both directly and indirectly interfere with fatty acid synthesis, an optimal means to assess their efficacy is lacking.  Given that ¹¹C-acetate is directly linked to the fatty acid synthesis pathway, this probe provides a unique opportunity to monitor lipogenic tumors by PET.  Herein, we review the relevance of the fatty acid synthesis pathway in cancer.  Furthermore, we address the potential utility of ¹¹C-acetate PET in imaging tumors, especially those that are not FDG-avid.  Last, we discuss several therapeutic interventions that could benefit from ¹¹C-acetate PET to monitor therapeutic response in patients with certain types of cancers.
[Back to top]


Regulation of Glycolytic and Mitochondrial Metabolism by Ras
J. Chesney and S. Telang
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00267]

High glucose uptake is a characteristic of most metastatic tumors and activation of Ras signaling in immortalized cells increases glycolytic flux into lactate, de novo nucleic acid synthesis and the tricarboxylic acid cycle, and increases NADH shuttling, oxygen consumption and uncoupling of ATP synthase from the proton gradient.   Fructose-2,6-bisphosphate, C-Myc, HIF1α and AKT each have been found to be key regulators of glycolysis and to be controlled by Ras signaling, and there is abundant evidence for cross-talk between these regulators.  The reprogramming of glycolytic and mitochondrial metabolism by Ras enables an integrated activation of energetic and anabolic pathways via the redox state of NADH that is required for the survival and growth of neoplastic cells in poorly vascularized tumors.  Several small molecule antagonists specific for essential metabolic enzymes have been found to be selectively toxic to Ras-transformed cells as opposed to wild-type cells, indicating that this metabolic reprogramming and addiction may have utility for the development of anti-neoplastic agents.
[Back to top]


History of avermectin and ivermectin, with notes on the history of other macrocyclic lactone antiparasitic agents
William C. Campbell
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00269]

The macrocyclic lactones enjoy a position of prominence in the control of parasites, and their history may be of interest, and even of use, in an age in which the search for chemotherapeutic agents has been transformed by modern technology.  Much of their history has been recorded piecemeal in a wide variety of publications.  The present review provides additional detail, and offers a personal perspective on the history of ivermectin and related avermectins.  Brief notes are included on the subsequent development of other macrocyclic lactones.   Milbemycin preceded the avermectins as a macrocyclic lactone of agricultural importance, but was used for a different purpose.  Development of the avermectins arose from the isolation, in the laboratories of the Kitasato Institute, of a novel soil-dwelling bacterium and its transmittal (in 1974) to the laboratories of Merck & Co., Inc.  There it was found (in 1975) to produce a potent anthelmintic substance, which was then identified and transmuted by interdisciplinary research into an antiparasitic product.  Initially the focus was on its applicability to veterinary science and animal husbandry; and after developmental research by many scientific teams, the product was introduced commercially (in 1981) for the control of endoparasitic nematodes and ectoparasitic arthropods in livestock.  Subsequently, special applications in human medicine were developed, and were successfully implemented in partnership with World Health Organization and several non-governmental organizations  (NGOs).
[Back to top]


Macrocyclic Lactone Anthelmintics: Spectrum Of Activity And Mechanism Of Action
Timothy G. Geary and Yovany Moreno
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00270]

Macrocyclic lactones (MLs), exemplified by the prototype of the class, ivermectin (IVM), are mainstays of programs for the control of nematode and arthropod parasites and pests. Since their introduction 30 years ago, research has revealed that they act on a family of ligand-gated chloride channels gated by glutamate, which is largely restricted to animals in the phyla Nematoda and Arthropoda. Studies on IVM in model organisms have contributed greatly to our understanding of ML pharmacology, but our understanding of the basis for differences among species and among MLs in potency and spectrum remains far from complete.
[Back to top]


Resistance to Macrocyclic Lactones
Adrian J Wolstenholme and Ray M. Kaplan
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00271]

Resistance to the macrocyclic lactones (MLs) has been confirmed or suspected in many target organisms and is a serious problem in some. For some species, such as parasitic nematodes of small ruminants, ML resistance has become severe enough to threaten effective worm control worm control. Resistance is also a major concern in horse parasites and an emerging problem in cattle. Despite this, we have insufficient understanding of the mechanisms of ML resistance, especially in nematodes. Some insect and mite agricultural pests express higher levels of detoxifying enzymes, leading to cross-resistance to other pesticide classes. A major difficulty is in the identification of true resistance and distinguishing this from other causes of treatment or prophylaxis failure – some in vitro assays for ML resistance are available but more are badly needed. Changes in the way anthelmintic drugs are used in livestock farming have been proposed, based on the treatment of those animals that would benefit most, and schemes have been devised for identifying those animals, flocks and herds. The continued sustainable use of these invaluable drugs may depend on the adoption and improvement of such schemes, as resistance is likely to become an ever more serious problem.
[Back to top]


Pharmacokinetic features of the antiparasitic Macrocyclic Lactones
Quintin A McKellar and Cengiz Gokbulut
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00272]

The macrocyclic lactones have pharmacokinetic properties which enhance their use against endo- and ectoparasites in animals and man. The most consistent physico-chemical feature of the group which contributes to their kinetic characteristics is high lipid solubility. This appears to be necessary for their pharmacodynamic action as well as common kinetic features such as large volumes of distribution and the influence of body fat composition on their disposition.

They are used in all domestic animal species and are undoubtedly influenced by the anatomical and physiological differences in these species, however body fat composition also appears to exert a major influence on distribution, metabolism and persistence between species and between breeds and individuals. A myriad of formulations have been developed to enhance the convenience of administration in the different domestic animals and the macrocyclic lactones are delivered orally, subcutaneously and topically to good effect. Lipid based excipients have been developed in “depot” formulations to extend the period of effective prevention of parasite re-infection.

Subtle structural changes have been made to the macrocyclic lactone molecules to reduce distribution to the central nervous system and mammary gland, thus allowing use of some compounds such as selamectin (SLM) in “toxicity sensitive” breeds of collie dog which lack P-glycoprotein efflux systems in their central nervous systems and the use of eprinomectin (EPM) in dairy cattle with a nil-milk withdrawal period.

Gender differences exist in the pharmacokinetics of these compounds which may be associated with body (fat) composition or metabolism. Feeding may also reduce the availability of macrocyclic lactones which bind particulate digestive material and parasitism may impact the kinetics of the drugs because parasitized animals have altered pathophysiological processes, especially in the gastro intestinal tract but also because of the impact which parasitism may have on the body condition (and fat deposition) in animals.

The pharmacokinetics of macrocyclic lactones may be affected by co-administration with compounds which interfere with P-glycoprotein transporters and these interactions have been explored as possible mechanisms for enhancing the effectiveness of these antiparasitics.

The objective of this article is to provide a comprehensive review of the pharmacokinetics of macrocyclic lactones and to interpret where that information may prove clinically useful.
[Back to top]


Macrocyclic lactones and cellular transport-related drug interactions -A perspective from in vitro assays to nematode control in the field-
Lifschitz, A. Ballent, M. Lanusse, C.
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00273]

Macrocyclic lactones (MLs) are antiparasitic drugs used against endo-ectoparasites. Regarding the wide use of MLs in different species, it is likely that drug-drug interactions may occur after their co-administration with other compounds. A new paradigm was introduced in the study of the pharmacology of MLs during the last years since the interactions of MLs with ATP-binding cassete (ABC) transporters have been described. The current review article gives an update on the available information concerning drug-drug interactions involving the MLs. The basis of the methodological approaches used to evaluate transport interactions, and the impact of the pharmacology-based modulation of drug transport on the MLs disposition kinetics and clinical efficacy, are discussed in an integrated manner.

A different number of in vitro and ex vivo methods have been reported to study the characterization of the interactions between MLs and ABC transporters. The production of the ABC transporters knockout mice has provided valuable in vivo tools to study this type of drug-drug interaction. In vivo trials performed in different species corroborated the effects of ABC transporter modulators on the pharmacokinetics behaviour of MLs. Important pharmacokinetic changes on plasma disposition of MLs have been observed when these compounds are co-administered with P-glycoprotein modulators. The modulation of the activity of P-glycoprotein was evaluated as a strategy not only to increase the systemic availability of MLs but also to improve their clinical efficacy. The understanding of the MLs interactions may supply relevant information to optimize their use in veterinary and human therapeutics.
[Back to top]


Extra-Label Use of Ivermectin in Some Minor Ruminant Species: Pharmacokinetic Aspects
González-Canga, A. Belmar-Liberato, R. Escribano, M.
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00274]

The characterisation of ivermectin pharmacokinetics can be used to predict and to ensure an optimal activity in the target species and for designing programmes aimed for parasite control. Ivermectin pharmacokinetic studies performed in several minor ruminant species are reviewed in this paper with the aim of facilitating the adoption of rational basis for the establishment of appropriate dosage schedules.
[Back to top]


Recent developments in the analysis of avermectin and milbemycin residues in food safety and the environment
Martin Danaher, Wolfgang Radeck, Lucija Kolar, Jemma Keegan and Vesna Cerkvenik-Flajs
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00275]

A review of the developments on the analysis residues of avermectins and milbemycins (both macrocyclic lactones) is presented.  The macrocyclic lactones (MLs) are an important class of chemicals, which are used worldwide as veterinary drugs and as crop protection agents.  As a result, residues of MLs are important from both a food safety and environmental perspective.  A review of the developments in ML residues in food was carried out in detail in 2006.  As a result, this paper covers recent developments in the area of food analysis, which are mainly multi-residue assays based on liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS).  A brief coverage of HPLC fluorescence (HPLC-FLD) based methods is included for completeness.  The paper will carry out a comprehensive review of ML residues in environmental samples.  These additional sections are reflective of the growing number of research papers published on LC-MS/MS and environmental applications in recent years.
[Back to top]


Toxicity in Animals: Target Species
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00276]
K N Woodward

The macrocyclic lactone endectocides typified by ivermectin are safe and effective drugs when used according to label directions. However, off-label use, misuse and overdosing can result in toxicity in animal patients as revealed by pharmacovigilance activities. Preclinical toxicity studies demonstrates that the major clinical signs of toxicity are those associated with neurotoxic effects and these are the most common adverse drug reactions noted in overdosed treated animals. Subpopulations of some strains or breeds of some species appear to be uniquely sensitive to the neurotoxic effects of the macrocyclic lactones due to enhanced brain penetration by these drugs as a result of a deficiency in P-glycoprotein arising as a result of a mutation in the MDR1 gene.
[Back to top]


Treatment of MDR1 mutant dogs with macrocyclic lactones
Joachim Geyer and Christina Janko
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00277]

P-glycoprotein, encoded by the multidrug resistance gene MDR1, is an ATP-driven drug efflux pump which is highly expressed at the blood-brain barrier of vertebrates. Drug efflux of macrocyclic lactones by P-glycoprotein is highly relevant for the therapeutic safety of macrocyclic lactones, as thereby GABA-gated chloride channels, which are confined to the central nervous system in vertebrates, are protected from high drug concentrations that otherwise would induce neurological toxicity. A 4-bp deletion mutation exists in the MDR1 gene of many dog breeds such as the Collie and the Australian Shepherd, which results in the expression of a non-functional P-glycoprotein and is associated with multiple drug sensitivity. Accordingly, dogs with homozygous MDR1 mutation are in general pone to neurotoxicity by macrocyclic lactones due to their increased brain penetration. Nevertheless, treatment of these dogs with macrocyclic lactones does not inevitably result in neurological symptoms, since, the safety of treatment highly depends on the treatment indication, dosage, route of application, and the individual compound used as outlined in this review. Whereas all available macrocyclic lactones can safely be administered to MDR1 mutant dogs at doses usually used for heartworm prevention, these dogs will experience neurological toxicity following a high dose regimen which is common for mange treatment in dogs. Here, we review and discuss the neurotoxicological potential of different macrocyclic lactones as well as their treatment options in MDR1 mutant dogs.
[Back to top]


Ivermectin Residue Depletion in Food Producing Species and its Presence in Animal Foodstuffs With a View to Human Safety
Escribano, M. San Andrés, M.I. de Lucas, J.J. González-Canga, A.
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00278]

From a human safety perspective, the administration of ivermectin to food producing animal species entails potential risks related to the presence of drug residues in edible tissues, milk, and other derived products. The European Medicines Agency has established the maximum residue limits for ivermectin in the European Union, with values of 100 μg∙kg^-1 in fat and liver and 30 μg∙kg^-1 in kidney for all mammalian food producing species, in order to ensure that the amount of ivermectin that can be found in animal foodstuff is below dangerous levels for the consumers. According to these values, withdrawal periods after subcutaneous injection were recently established in the European Union (2009), in 49 days for products containing ivermectin as a single active substance or in combination with closantel, and in 66 days when combined with clorsulon. The marker residue for ivermectin was found to be H₂B_1a, which is the major component of the parent compound. The tissue distribution of residues and the overall ratios of marker to total residues were generally similar in most species, and the highest concentrations of ivermectin residues were found in fat and liver with high levels also detected in injection site muscles. Ivermectin is not licensed for use in animals from which milk is produced for human consumption, however its extra-label use should be considered regarding human safety, due to its long persistence in milk and milk-derived products.
[Back to top]


Acute Human Toxicity of Macrocyclic Lactones
Chen-Chang Yang
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00279]

Macrocyclic lactones, including avermectins and milbemycins, are novel parasiticides and insecticides that are produced through fermentation by soil-dwelling microorganisms. Although various macrocyclic lactones may differ in their potency and safety, all of them are believed to share common pharmacologic/toxicologic mechanisms, i.e. leading to paralysis and death of parasites and other target organisms via the activation of a glutamate-gated chloride channel in the invertebrate nerve and muscle cells and/or through the effect on gamma-aminobutyric acid (GABA) receptors. Ivermectin is the first macrocyclic lactone that was released for use in both animals and humans, and has demonstrated both excellent efficacy and high tolerability in the treatment of parasite infestations. Other macrocyclic lactones, such as abamectin, emamectin, and moxidectin were subsequently commercialized and have been used as insecticides and acaricides for crop protection or parasiticides for animal health.

Although ivermectin therapy is generally well tolerated, adverse effects that are usually transient and mild-to-moderate can occur. Severe adverse effects are rare and can generally be effectively controlled by symptomatic measures. Non-therapeutic exposures to ivermectin and other macrocyclic lactones may also result in toxic effects; significant toxicity however probably develops only after large amount of oral ingestion. Although the exact mechanisms remain unclear, macrocyclic lactones in large doses may pass through the BBB to produce GABA-mimetic toxic effects. Severely poisoned patients usually present with coma, hypotension, respiratory failure, and even death. Despite the lack of specific therapy, the prognosis is likely to be favorable unless the poisoned patients are complicated with severe hypotension or respiratory failure.
[Back to top]


A review on the toxicity and non-target effects of macrocyclic lactones in terrestrial and aquatic environment
Jean-Pierre Lumaret, Faiek Errouissi, Kevin Floate, Jörg Römbke, Keith Wardhaugh
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00280]

The avermectins, milbemycins and spinosyns are collectively referred to as macrocyclic lactones (MLs) which comprise several classes of chemicals derived from cultures of soil micro-organisms. These compounds are extensively and increasingly used in veterinary medicine and agriculture. Due to their potential effects on non-target organisms, large amounts of information on their impact in the environment has been compiled in recent years, mainly caused by legal requirements related to their marketing authorization or registration. The main objective of this paper is to critically review the present knowledge about the acute and chronic ecotoxicological effects of MLs on organisms, mainly invertebrates, in the terrestrial and aquatic environment. Detailed information is presented on the mode-of-action as well as the ecotoxicity of the most important compounds representing the three groups of MLs.

This information, based on 360 references, is mainly provided in nine tables, presenting the effects of abamectin, ivermectin, eprinomectin, doramectin, emamectin, moxidectin, and spinosad on individual species of terrestrial and aquatic invertebrates as well as plants and algae. Since dung dwelling organisms are particularly important non-targets, as they are exposed via dung from treated animals over their whole life-cycle, the information on the effects of MLs on dung communities is compiled in an additional table. The results of this review clearly demonstrate that regarding environmental impacts many macrocyclic lactones are substances of high concern particularly with larval instars of invertebrates. Recent studies have also shown that susceptibility varies with life cycle stage and impacts can be mitigated by using MLs when these stages are not present. However information on the environmental impact of the MLs is scattered across a wide range of specialised scientific journal with research focusing mainly on ivermectin and to a lesser extent on abamectin, doramectin and moxidectin. By comparison, information on compounds such as eprinomectin, emamectin, selamectin is still relatively scarce.
[Back to top]


Use of Macrocyclic Lactones in Cattle in the USA
Lora R. Ballweber and Laurie A. Baeten
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00281]

The use of macrocyclic lactones has become the main stay for the treatment of endo- and ectoparasites in the cattle industry.  Here we review those drugs that are currently approved for use in cattle in the United States.  The general efficacy, tissue distribution and toxicity of each drug formulation are discussed.  Included is a discussion regarding the current status for nematode anthelmintic resistance in cattle populations within the United States.  Also provided is a current summary of ecological effects of macrocyclic lactones residues in manure.
[Back to top]


Macrocyclic Lactones for Parasite Control in Equids
E. T. Lyons and S. C. Tolliver
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00282]

Macrocyclic lactones (MLs) revolutionized parasite control in horses and other animals.  They are unique in that they are effective against arthropods and nematodes. The first of the widely used avermectins was ivermectin.  In 1983, it was marketed for use in horses as an injectable formulation but was withdrawn in 1984 after about a year and half on the market because of adverse problems.  It was replaced by a paste formulation and an oral/stomach tube liquid formulation.  Ivermectin is highly active on bots, ascarids, large and small strongyles, pinworms, strongyloides, stomach worms, and some other internal parasite species.  Another ML, moxidectin, became available in 1997 as a gel formulation for oral administration.  The parasiticidal activity of this compound is similar to ivermectin except efficacy is less on the common bot (Gasterophilus intestinalis) but high on encysted small strongyles.  Recently however lower than initial activity on ascarids and small strongyles has been found for both ivermectin and moxidectin.
[Back to top]


Macrocyclic Lactones in the Treatment and Control of Parasitism in Small Companion Animals
Thomas J. Nolan and James B. Lok
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00283]

Macrocyclic lactones (MLs) have many anti-parasitic applications in small companion animal medicine.  They were first developed as chemoprophylactics against heartworm (Dirofilaria immitis) infection to be applied monthly for retroactive killing of third- and fourth-stage larvae.  ML-containing products formulated for oral (ivermectin, milbemycin oxime), topical (selamectin, moxidectin) or injectable sustained release (moxidectin, ivermectin) are approved for heartworm prevention in dogs or cats.  Clearance of microfilariae and gradual or “soft” killing of adult heartworms constitute increasingly prevalent extra-label uses of MLs against D. immitis. Some commercial ML formulations contain sufficient levels of active ingredient (milbemycin oxime, selamectin, moxidectin) to support additional label claims against gastrointestinal nematode parasites such as hookworms (Ancylostoma spp.) and ascarid round worms (Toxocara spp. and Toxascaris leonina). Beyond these approved applications, safe, extra-label uses of MLs against nematodes parasitizing the urinary tract, such as Capillaria spp., and parasites of the tissues, such as Dipetalonema reconditum, Dirofilaria repens, Thelazia spp. and Spirocerca lupi, in dogs and cats as well as exotic pets have been reported.  MLs as a group have intrinsic insecticidal and acaricidal activity, and topical or otic formulations of certain compounds (selamectin, moxidectin, milbemycin oxime or ivermectin) are approved for treatment and control of fleas, certain ixodid ticks, sarcoptiform and demodectic mange mites and psoroptiform ear mites.  Extra-label applications of MLs against ectoparasites include notoedric mange mites, dermanyssids such as Ornythonussus bacoti, numerous species of fur mite (e.g. Cheyletiella spp. and Lynxacarus) and trombiculids (“chiggers”) in cats, dogs and nontraditional or exotic pets.
[Back to top]


Avermectin Use In Aquaculture
Tor E. Horsberg
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00284]

The main indication for use of avermectins in aquaculture-produced fish is infestations with ectoparasitic copepods. The compounds ivermectin and emamectin benzoate are predominantly used as in-feed formulations on salmonid fish against copepods in the family Caligidae: Lepeophtheirus salmonis, Caligus elongatus  and C. rogercresseyi. These agents are well-documented as very effective on all developmental stages of the parasites. The duration of effect can be up to 10 weeks. The safety margin for ivermectin is narrow, but better for emamectin benzoate. Environmental impact from these chemicals on bottom-dwelling and sediment-dwelling organisms occurs, but these are restricted to the immediate area around the production site. Avermectins are incompletely absorbed from the intestine of the fish and slowly excreted. They penetrate the blood-brain barrier of the fish, ivermectin more than emamectin benzoate. Resistance has developed against these agents in L. salmonis in almost all major salmon producing areas. The situation must be viewed as serious and can render these agents completely ineffective for salmon lice control.
[Back to top]


Ivermectin in human medicine, an overview of the current status of its clinical applications
González, P. González, F.A. Ueno, K.
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00285]

Ivermectin is a broad spectrum antiparasitic veterinary drug introduced in human medicine in 1987. It is considered the drug of choice in onchocerciasis and strongyloidiasis infections, and remains as a therapeutic option for mass treatment in lymphatic filariasis, for which it has widely proved its efficacy. While research continued for human use, new therapeutic targets for ivermectin have emerged. It is currently the better therapeutic option in the treatment of gnathostomiasis and crusted scabies, and could be an alternative option in ascariasis and Mansonella infections. Although these uses are already included in clinical guidelines, more trials are needed to increase their grade of evidence and to obtain their official approval. Concerning other minor uses such as the treatment of enterobiasis or against Trichuris trichiura, more research is still needed in order to test the real activity of ivermectin. The use of ivermectin in human medicine has shown an outstanding low rate of adverse reactions, with the exception of treatment of loiasis and onchocerciasis, where the death of a high microfilarial load may cause severe encephalopathy. However special attention must be paid to the emergence of the first documented cases of resistance in treatment of scabies.
[Back to top]


Beyond providing drugs, the mectizan ® donation creating paradigm shifts in health care and health systems strengthening
Dr Adrian Hopkins
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00285]

The donation of Mectizan®   by Merck & Co Inc. in 1987 “as much as was needed for as long as was needed for onchocerciasis control” was a major change from traditional corporate drug donations.  The company realised that those who needed the drug most would never be able to purchase it, and so gave it away. The donation enabled the Onchocerciasis Control Programme in West Africa to add Mectizan distribution to its ongoing control strategy. For the first time there was hope for those living in other areas of Africa, Latin America and Yemen. Governments and non-governmental development organizations quickly got together to begin treatment in these new areas. Two new programmes and partnerships were created; the African Programme for Onchocerciasis Control and the Onchocerciasis Elimination Programme for the Americas. These programmes have been in the forefront of developing new strategies, including the Community Directed approach, which has now expanded into other disease control programmes at the community level, such as Vitamin A distribution and malaria control. This donation has led not only  to the probability of elimination of onchocerciasis in the Americas in the near future, but is stimulating approaches to the elimination in Africa, in areas considered impossible five years ago. Other major pharmaceutical donations have followed, initiating the plan to eliminate lymphatic filariasis worldwide, and also stimulating interest in controlling other “neglected tropical diseases,” which affect the poorest billion of the world’s population, making this now a reality.
[Back to top]


Image analysis of colocalization of nuclear DNA and GFP labelled HIF-1α in stable transformants
Takehito Goto, Michihiko Sato, Eiji Takahashi and Yasutomo Nomura
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00287]

HIF-1a is regarded as a target for drug development in several diseases such as cancer.  For high throughput screening of HIF-1α-targeted drug, we need to examine the activity quantitatively.In the present study, we proposed a method where stable expression system of HIF-1awas combined with image correlation analysis.  When the stable transformants were labeled with DRAQ5, we could detect Co²⁺-induced nuclear translocation by the use of cross-correlation analysis of the dual labeling images.  In the case of high throughput screening for HIF-1α-targeted drug, we should use Pearson’s correlation coefficient to judge nuclear translocation.
[Back to top]


The reaction mechanism of calcium-activated photoprotein bioluminescence
Takashi Hirano
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00288]

Calcium-activated photoproteins are important and useful bioluminescent reagents for detecting the calcium ion (Ca²⁺) in biological systems. In conjunction with photon imaging technology, they can be used to observe Ca²⁺-related life processes in a living cell. To develop useful applications of calcium-activated photoproteins, we need to understand the molecular basis of the bioluminescence reaction. For this purpose, this review describes the oxygenation, chemiexcitation, and light emission processes of calcium-activated photoproteins in the bioluminescence reaction together with the fundamental chemistry of the luminous substrate, coelenterazine, based on recent results from mechanistic chemical studies of these primary processes. Finally, the whole reaction mechanism, including the active site structures of apoproteins, along with available information about the molecular mechanism and the crystallographic structures of calcium-activated photoproteins are summarized.
[Back to top]


Low-Coherence Dynamic Light Scattering and its Potential for Measuring Cell Dynamics
Katsuhiro Ishii
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00289]

Fluorescence correlation spectroscopy is a powerful technique for studying the structures and dynamics of living cells.  Dynamic light scattering (DLS) is also used to study dynamic characteristics and it has the potential to measure cell dynamics.  However, it is difficult to apply DLS to highly scattering media.  In this article, we review low-coherence dynamic light scattering (LC-DLS).  It strongly suppresses the influence of multiple scattering and has a greater potential for measuring cell dynamics than conventional DLS.  The properties of LC-DLS are described theoretically and experimentally.  Measurement of the diffusion coefficients of macromolecules in turbid media and interparticle and molecular interactions by LC-DLS is demonstrated.
[Back to top]


A Novel Molecular Design Strategy For Efficient Two-Photon Absorption Materials
Jun Kawamata and Yasutaka Suzuki
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00290]

This paper describes a novel molecular design strategy for obtaining efficient two-photon absorption (TPA) materials. The most popular strategy for enhancing the TPA cross-section (σ⁽²⁾) of a molecule is to enhance its transition dipole moment. However, this strategy also red shifts the one-photon absorption (OPA) band. Consequently, molecules with large transition dipole moments typically exhibit strong OPA at visible wavelengths, making it difficult to use such molecules for TPA-related applications in the visible wavelength region. Therefore, an alternative molecular design principle for TPA materials to enhance the transition dipole moment is strongly required. The present paper describes a novel molecular design strategy for reducing the detuning energy by incorporating an azulenyl moiety in a large, planar π-electron system. This strategy enhances σ⁽²⁾ without significantly red shifting the OPA band.
[Back to top]


Magnetically-Modulated Atomic Force Microscopy for Analysis of Soft Matter Systems
Masami Kageshima
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00291]

Experimental method of studying viscoelasticity, a common idea to understand properties of microscopic biological soft matter systems, especially single biopolymer chains, using atomic force microscopy (AFM) with magnetically-driven cantilever is surveyed. The experimental setup of applying well-characterized excitation to the cantilever and the analysis method to derive the viscoelasticity of the system under study are briefly introduced. Examples of measuring viscoelasticity of single peptide molecule and single titin molecule are shown. Considering the close relation of viscoelasticity and the time-scale for nonequilibrium dynamics in soft matter, extension of the method to a frequency-resolved analysis is attempted. A result of measuring viscoelasticity spectrum of a single dextran chain is shown. Challenges in further progress of the method are also described.
[Back to top]


Single Molecular observation of DNA and DNA Complexes by Atomic Force Microscopy
Takuya Matsumoto
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00292]

Atomic force microscopy (AFM) provides a novel way to understand the structure-function relationship of protein synthesis at a single-molecular level. High-resolution AFM imaging in air, liquid and vacuum allows for single DNA, RNA and proteins to be observed at the nano-scale in addition to their conformational transitions, bound states, temporal behavior, and assembly. The recent development of frequency modulation mode AFM has led to imaging hydration structures of DNA in water and molecular polarization of DNA complexes in vacuum. Real-time imaging in near-physiological environments captures transcriptional activation with characteristic conformation of DNA-protein complexes. We review current achievements and the future potential of methodological and biological AFM to reveal insights into DNA, RNA and their complexes.
[Back to top]


Effect of Isoproterenol on Local Contractile Behaviors of Rat Cardiomyocytes Measured by Atomic Force Microscopy
Takaharu Okajima
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00293]

Inotropic agents induce changes in the contraction amplitude and frequency of cardiomyocytes (CMs). However, it is unknown how local contractions of CMs treated by inotropic agents behave spatiotemporally. In this study, the effect of isoproterenol, a positive inotropic agent, on local contractions of isolated neonatal rat CMs was explored by atomic force microscopy (AFM). We observed that changes in local contraction amplitude of CM in the presence of isoproterenol were heterogeneous; they were unchanged or increased, at different positions, with respect to the amplitude of untreated CMs. Interestingly, spatial heterogeneities of local contraction amplitude of CM in the presence of isoproterenol did not obviously correlate with the local elasticity, indicating that the local contractions were facilitated by cooperative dynamics of the cytoskeletal structure in relatively large regions, rather than those just under AFM indentation. Moreover, local contraction amplitude of CM in the presence of isoproterenol was not proportional to that in the control condition, showing that the former change was no longer additive in local scales.
[Back to top]


Current Research on Protein-Protein Interactions Among Auxin-Signaling Factors in Regulation of Plant Growth and Development
Hideki Muto and Masataka Kinjo
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00294]

In many signaling pathways, various factors have been isolated by molecular genetic studies and whole-genome analysis.  Understanding the interactions among these factors is crucial to understanding the signaling process as a whole.  Recent studies on auxin signaling in the regulation of plant growth and development have discovered primary factors interacting with each other, and elucidated a very simple pathway modulating gene expression in response to auxin.  However, these studies of auxin-signaling led to the question of how such a simple pathway generates multiple types of regulation in various processes of different cells throughout the life of a plant.  Here, we provide an overview of recent progress in auxin biology focusing on protein-protein interactions in the signal transduction pathway and discuss various possibilities and approaches to resolve the issue.
[Back to top]


Use of Carbohydrate-Conjugated Nanoparticles for an Integrated Approach to Functional Imaging of Glycans and Understanding of Their Molecular Mechanisms
Noriko Nagahori, Makiyo Uchida, Masataka Kinjo and Tadashi Yamashita
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00295]

Functional analysis of carbohydrates is needed to understand the initial interface between membranes and the outer world. For this analysis we need individual protocols such as a method to modify the surfaces of nanoparticles with a variety of carbohydrates effectively and exhaustively, to synthesize an oligosaccharide on each particle’s surface by chemical or enzymatic sugar elongation reaction, and to analyze the binding properties of carbohydrates using surface plasmon resonance (SPR). Moreover, to elucidate the functions of carbohydrates in vivo, we attempted real-time live cell / animal imaging using quantum dots (QDs) conjugated with carbohydrates. In this article, we describe the basic strategies for scooping up proteins from crude sample mixtures via interaction with carbohydrates. This approach was used to identify proteins that interacted with GM2, a ganglioside that is abundant on the surfaces of human lung cancer cells.
[Back to top]


Direct Quantification of Mitochondria and Mitochondrial DNA Dynamics
Dr. Yasutomo Nomura
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00296]

Mitochondria are known to be one of major organelles within a celland to play a crucial role in many cellular functions.  These organelles show the dynamic behaviors such as fusion, fission and the movement along cytoskeletal tracks.  Besides mitochondria, mitochondrial DNA is also highly motile.  Molecular analysis revealed that several proteins are involved in mitochondria and mitochondrial DNA dynamics.  In addition to the degeneration of specific nerves with high energy requirement, mutation of genes coding these proteins results in metabolic diseases.  During the last few years, a significant amount of relevant data has been obtained on molecular basis of these diseases but mitochondrial dynamics in cells derived from the patients is poorly understood.  So far time-lapse fluorescence microscopy, fluorescence recovery after photobleaching and image correlation methods have been used to study organellar motion.  Especially, image correlation method has possibility to evaluate diffusion coefficient mitochondria and mitochondrial DNAsimultaneously and directly.  When we search candidates for compounds that modulate mitochondrial dynamics by high throughput screening, image correlation methodmay be useful although the careful interpretation is required for crowded and heterogeneous environment within a cell.
[Back to top]


Atomic Force Microscopy for the Examination of Single Cell Rheology
Takaharu Okajima
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00297]

Rheological properties of living cells play important roles in regulating their various biological functions. Therefore, measuring cell rheology is crucial for not only elucidating the relationship between the cell mechanics and functions, but also mechanical diagnosis of single cells. Atomic force microscopy (AFM) is becoming a useful technique for single cell diagnosis because it allows us to measure the rheological properties of adherent cells at any region on the surface without any modifications. In this review, we summarize AFM techniques for examining single cell rheology in frequency and time domains. Recent applications of AFM for investigating the statistical analysis of single cell rheology in comparison to other micro-rheological techniques are reviewed, and we discuss what specificity and universality of cell rheology are extracted using AFM.
[Back to top]


Current Status and Future Prospects for Research on Tyrosine Sulfation
Nobuya Sasaki
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00298]

Tyrosine (Tyr) sulfation is a common posttranslational modification of secreted proteins or membrane-bound proteins that is implicated in numerous physiological and pathological processes. The Tyr sulfation modifies protein-protein interactions involved in leukocyte adhesion, homeostasis, and receptor-mediated signaling. To data, 80 Tyr-sulfated proteins have been identified. As new methodologies and bioinformatics for the detection of Tyr sulfation become available, the number of Tyr-sulfated acceptor proteins discovered is bound to increase. Further, recent advances in microscopy and fluorescence technology will provide information on the true spatial and temporal nature of Tyr-sulfated proteins within the intact cell. This review summarizes the methods for the detection of Tyr O-sulfation as well as the biological functions of sulfated Tyr. Further, illustrative examples of the impact of Tyr sulfation on the pharmacological properties are presented.
[Back to top]


Synthesis and Application of Visible Light Sensitive Azobenzene
Shinjiro Sawada, Nobuo Kato and Kunihiro Kaihatsu
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00299]

Methods for regulating peptide conformation by non-harmful light stimuli can be useful for remotely controlling cellular functions in vitro. Here, we synthesized a series of p-heteroatom-substituted azobenzenes and studied their photoisomerization properties. The trans-isomer of p-sulfur-substituted azobenzene was effectively isomerized by visible light irradiation and the cis-isomer was thermally stable at physiological temperature. We developed a novel visible light sensitive amino acid (AZO), via p-sulfur-substituted azobenzene, and utilized it as a photosensitive modulator of the SV40 nuclear localization signal (NLS). The cellular uptake of the AZO-NLS conjugate was controlled by visible light irradiation. Our technology can be utilized for regulating not only the cellular uptake, but also the function of peptides within cells by non-harmful visible light irradiation.
[Back to top]


Fluorescence Imaging of Mitochondria in Living Cells Using a Novel Fluorene Derivative with a Large Two-Photon Absorption Cross-Section
Seiji Tani, Kenta Nakagawa, Takuya Honda, Hiromasa Saito, Yasutaka Suzuki, Jun Kawamata, Makiyo Uchida, Akira Sasaki and Masataka Kinjo
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/0300]

We have examined optical properties of a fluorene derivative with two positively charged substituents, 1,1’-diethyl-4,4’-(9,9-diethyl-2,7-fluorenediyl-2,1-ethane­diyl)dipyridi­nium perchlorate (1), in water.  The photoluminescence quantum yield of 1 was relatively high (35%) for use as a fluorescent probe in water.  We also examined two-photon absorption (TPA) properties of 1 in methanol.  The maximum value of the TPA cross-section (730 GM at 750 nm, 1 GM = 10^-50 cm⁴ s photon^-1 molecule^-1) was larger than that for most two-photon-excited fluorescent dyes including a classical mitochondria-selective fluorescent dye rhodamine 123.  Preliminary fluorescence imaging experiments of the mitochondria in Paramecium caudatum and Hela living cells were carried out with 1.  Bright green fluorescence was observed from the mitochondria in both living cells loaded 1 without toxicity effects.  These our results indicate that water-soluble fluorene derivative 1 is a promising candidate as a two-photon-excited fluorescence probe for mitochondria in living cells.
[Back to top]


Highly Controllable Optical Tweezers Using Dynamic Electronic Holograms
Johtaro Yamamoto and Toshiaki Iwai
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00301]

Dielectric particles including living cells are trapped within focused laser beam spots, and as a result, they can be transferred by displacing the beam spots.  Such the particle manipulating technique is called optical tweezers.  Holographic optical tweezers (HOT) enables highly flexible and precise control of particles, introducing holography technique to them.  HOT is one of the most expected techniques for investigations of cell-cell signaling which require precise arraying of living cells.  We had developed a new highly controllable HOT system where two different intensity patterns, a carrier beam spot and a beam array, are generated quasi-simultaneously by time-division multiplexing.  Particles are transferred to the beam array by the carrier beam spot displaced in real time by phase shifting of holograms.  In this review, we introduce our work, the construction of the system, demonstration of manipulating particles and investigations of the spatio-temporal stability of trapped particles in our system.
[Back to top]


Recent Advances in the Study of Glycosphingolipids
Tadashi Yamashita
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00302]

Glycosphingolipids (GSLs) are present in all mammalian cell plasma membranes and intracellular membrane structures. They are especially concentrated in plasma membrane lipid domains that are specialized for cell signaling. Plasma membranes show typical structures called rafts and caveola domain structures, with large amounts of sphingolipids, cholesterol, and sphingomyelin in the cell membranes. Plasma membranes have two faces, many kinds of receptors for intercellular signal transducers such as GPI-anchored proteins on the exoplasmic faces of the rafts/caveolae and src family kinases on the cytosolic face. Thus they play a role in transmembrane signal transduction, following the phosphorylation of some substrates and gene expression. On the other hand, their functions have become clear through the study of gene-manipulated mice. For further advances, a visual method to display diversity of biological functions is necessary. For this purpose, the use of high-performance microscopes and live cell imaging technologies are useful for more detailed understanding.
[Back to top]



In silico search for drug targets of natural compounds
Lixia Yao
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00304]

Natural compounds represent a significant source for the development of novel medicines. Finding the target proteins for a natural compound is the most important step towards understanding its molecular mechanism for therapeutic usage. In fact, the search for target proteins could be considered the first step of the drug discovery and development pipeline. While experimental determination of compound-protein interactions remains very challenging, effective in silico approaches have been developed and have demonstrated appealing advantages, including their low-cost and capability to scale up easily. The goal of this article is to provide an introduction to in silico search for drug targets of natural compounds. I first review currently available natural compounds databases and human gene/protein databases, and the rapidly emerging databases for known drug-target interactions. These resources provide the 'materials' for in silico approaches and define the gold standard of 'positives' for evaluating them. I then introduce three classes of computational methods for target identification of natural compounds, namely molecular docking, quantitative structure-relationship relationship (QSAR) modeling, and data mining and integrative analysis. Use of these methods is explained using real examples, and the advantages and disadvantages of each method are compared. As these state-of-the-art methods continue to mature amid significant challenges, this field appears poised for a period of significant growth, with untold benefits to drug discovery and natural product development.
[Back to top]


Pharmacophore, QSAR, and Binding Mode Studies of Substrates ofHuman Cytochrome P450 2D6 (CYP2D6)using Molecular Docking and Virtual Mutations and an Application to Chinese Herbal Medicine Screening
Sui-Lin Mo, Wei-Feng Liu, Chun-Guang Li, Hai-Bin Luo, Rongshi Li and Shu-Feng Zhou
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00305]

The highly polymorphic human cytochrome P450 2D6 (CYP2D6) metabolizes about 25% of currently used drugs. In this study, we have explored the interaction of a large number of substrates (n = 120) with wild-type and mutated CYP2D6 by molecular dockingusing the CDOCKER module. Before we conducted the molecular docking and virtual mutations, the pharmacophore and QSAR models of CYP2D6 were developedand validated. Finally, we explored the interaction of a traditional Chinese herbal formula, Fangjifuling decoction, with CYP2D6 by virtual screening. The optimized pharmacophore model derived from 20 substrates of CYP2D6 contained two hydrophobic featuresand one hydrogen bond acceptor feature, giving a relevance ratio of 76% when a validation set of substrates were tested. However, our QSAR models gave poor prediction of the binding affinity of substrates. Our docking study demonstrated that 117 out of 120 substrates could be docked into the active site of CYP2D6. Forty one out of 117 substrates (35.04%) formed hydrogen bonds with various active site residues of CYP2D6 and 53 (45.30%) substrates formed a strong π-π interaction with Phe120 (53/54), with only carvedilol showing π-π interaction with Phe483. The active site residues involving hydrogen bond formation with substrates included Leu213, Lys214, Glu216, Ser217, Gln244, Asp301, Ser304, Ala305, Phe483, and Phe484. Furthermore, the CDOCKER algorithm was further applied to study the impact of mutations of 28 active site residues (mostly non-conserved) of CYP2D6 on substrate binding modes using five probe substrates including bufuralol, debrisoquine, dextromethorphan, sparteine, and tramadol. All mutations of the residues examined altered the hydrogen bond formation and/or aromatic interactions, depending on the probe used in molecular docking. Apparent changes of the binding modes have been observed with the Glu216Asp and Asp301Glu mutants. Overall, 60 compounds out of 130 from Fangjifuling decoction matchedour pharmacophore modelfor CYP2D6 substrates. Fifty four out of these 60compounds could be docked into the active site of CYP2D6 and 24 of 54 compounds formed hydrogen bonds with Glu216, Asp301, Ser304, and Ala305 in CYP2D6. These results haveprovided further insights into the factors that determining the binding modes of substrates toCYP2D6. Screening of high-affinity ligands for CYP2D6 from herbal formula using computational models is a useful approach to identify potential herb-drug interactions.
[Back to top]


Integrated Analysis on the Physicochemical Properties of Dihydropyridine Calcium Channel Blockers in Grapefruit Juice Interactions
Yoshihiro Uesawa, Takahiro Takeuchi and Kiminori Mohri
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00306]

Concomitant consumption of grapefruit juice (GFJ) causes increases in the plasma concentration of a variety of drugs due to inhibition of intestinal CYP3A enzyme. Dihydropyridine calcium channel blockers belong to the category of drugs that are most prone to undergo such interaction. Increases inarea under the plasma concentration-time curve (AUC) due to GFJ differ greatly depending on the dihydropyridine administered. Therefore, a meta-analysis of each dihydropyridine was performed based on availableliterature. The criteria for using a publication were: subjects were healthy adults, dihydropyridinesweretaken with GFJ concomitantly or within one hour after intake of the juice, and the control groupadministered water in place of GFJ.In these studies, the investigations on GFJ interactions with 13 dihydropyridines such as amlodipine, azelnidipine, benidipine, cilnidipine, efonidipine, felodipine, manidipine, nicardipine, nifedipine, nimodipine, nisoldipine, nitrendipine and pranidipine were reported. As a result of meta-analyses, statistically significant interactions were not identified in amlodipine. Next, correlation analyses between the physicochemical properties and interaction strengths of the dihydropyridineswere performed to clarify the cause of the variation in the strengths that was dependent on the dihydropyridine. LogP, molecular weight, topological polar surface area (tPSA), molar refractivity, water diffusion, molecular volume, molecular density, molecular polarizability, and refractive index were calculated from the chemical structures. The interaction strength was defined as the logarithmic values of the increasing AUC ratio. The correlation analyses indicated a relationship of logP and tPSA with the interaction strengths. These findings suggest that the wide difference in the potency of interaction of each dihydropyridine may be explained by the presence of hydrophobic and electrostatic interactions between dihydropyridines and intestinal CYP3A enzyme.
[Back to top]


Herb-drug interactions: methods to identify potential influence of genetic variations in genes encoding drug metabolizing enzymes and drug transporters Running title: Herb-drug interaction pharmacogenetics
M. Hu, D. Q. Wang, Y. J. Xiao, V. W. L. Mak and B. Tomlinson
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00307]

Herbal supplements are often used concomitantly with conventional medications resulting in considerable potential for herb-drug interactions. These interactions, which are generally through interfering with pharmacokinetic and/or pharmacodynamic pathways, may result in beneficial effects or more often adverse reactions such as toxicity or treatment failure and may be influenced by multiple environmental and/or genetic factors. The pharmacogenetic approach may help to identify some interactions which may be more pronounced or only occur in specific groups of subjects although the complex nature of the herbal medicines may limit the discovery of such an interaction. Preclinical studies such as gene expression profiling in rodent liver may help to define metabolic pathways influenced by herbal medicines and facilitate more accurate targeting of human in vivo studies. This review discusses the mechanisms of herb-drugs interaction and the potential influence of genetic variation on herb-drug interactions based on available clinical evidence.

[Back to top]


Soy Isoflavones and Cardiovascular Disease Epidemiological, clinical and -omics perspectives
Gil-Izquierdo, A. Peñalvo, J.L Gil, J.I. Medina, S. Horcajada, M.N. Lafay, S. Silberberg, M. Llorach, R. Zafrilla, P. García-Mora and  P. Ferreres, F
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00309]

Cardiovascular disease (CVD) mortality rates are lower in Asian countries where dietary patterns are very different from Western diet. A number of studies have linked these lower rates to the inclusion of soy products as a staple food in those countries. Soy is the richest dietary source of isoflavones, a type of phytoestrogen associated with many potentially beneficial effects. Isoflavone-containing soy protein consumption has been linked to reduced levels of LDL cholesterol in hypercholesterolemic patients. This effect is increased with the concomitant administration of isoflavones, and seems to be also complemented by the isoflavone capacity to restore the endothelial function in patients with weak and moderated endothelial disfunction. The effects are variable depending on individuals’ metabolism and in particular to their ability to convert daidzein to equol that seems to be restricted to approximately 1/3 of the population. Equol production has been indeed linked to a decreased arterial stiffness and antiatherosclerotic effects via NO production. Because the relevance of isoflavones consumption on the modulation of cardiovascular risk still remains unclear, this paper aims to review the existing knowledge on the biological activity of the isoflavones on the human cardiovascular system from an epidemiological, clinical and –omics point of view.
[Back to top]


Plant secondary metabolites in cancer chemotherapy: where are we?
David M. Pereira, Patrícia Valentão, Georgina Correia-da-Silva, Natércia Teixeira and Paula B. Andrade
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00310]

Nowadays we have a number of chemical and biological agents at our disposal to treat chronic pathologies such as cancer. Although most drugs display significant activity, thus improving the clinical outcome, side-effects and emergence of resistances cannot be looked down.

From an historical point of view, higher plants have been very important in the search of new therapeutic agents and they were in the origin of the first medicines used in human health. The contribute of plants to treat pathologies such as cancer is far from being over, mainly due to the high number of new drugs that are currently being evaluated in clinical trials. Metabolomics-based studies have rendered several new chemical entities, some of them with remarkable complex chemistry, which sometimes results in novel mechanisms of action, higher potency and lower toxicity.

In this review, we will focus the most important plant-derived classes of compounds in clinical use, as well as those currently in clinical trials, with special focus on vinca alkaloids, taxanes, combretastatins, podophylotoxins and camptothecins . The molecular mechanism of action and spectrum of activity will also be discussed.
[Back to top]


Genomics and Cancer Drug Resistance
António S. Rodrigues, Joana Dinis, Marta Gromicho, Célia Martins, António Laires and José Rueff
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00311]

Cellular drug resistance is a major obstacle in cancer therapy. Mechanisms of resistance can be associated with altered expression of ATP-binding cassette (ABC) family of transporters on cell membrane transporters, the most common cause of multi-drug resistance (MDR), but can also include alterations of DNA repair pathways, resistance to apoptosis and target modifications.

Anti-cancer treatments may be divided into different categories based on their purpose and action: chemotherapeutic agents damage and kill dividing cells; hormonal treatments prevent cancer cells from receiving signals essential for their growth; targeted drugs are a relatively new cancer treatment that targets specific proteins and pathways that are limited primarily to cancer cells or that are much more prevalent in cancer cells; and antibodies function by either depriving the cancer cells of necessary signals or by causing their direct death. In any case, resistance to anticancer therapies leads to poor prognosis of patients. Thus, identification of novel molecular targets is critical in development of new, efficient and specific cancer drugs.

The aim of this review is to describe the impact of genomics in studying some of the most critical pathways involved in cancer drug resistance and in improving drug development. We shall also focus on the emerging role of microRNAs, as key gene expression regulators, in drug resistance. Finally, we shall address the specific mechanisms involved in resistance to tyrosine kinase inhibitors in chronic myeloid leukemia.
[Back to top]


Genomics and Pharmacogenomics of Brain Disorders
Ramón Cacabelos, Rocío Martínez-Bouza, Juan Carlos Carril, Lucía Fernández-Novoa, Valter Lombardi, Iván Carrera, Lola Corzo and Adam McKay
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00312]

CNS disorders are the third major problem of health in developed countries, with approximately 10% of direct costs associated with a pharmacological treatment of doubtful cost-effectiveness. There is an alarming abuse of psychotropic drugs worldwide and only 20-30% of patients with CNS disorders appropriately respond to conventional drugs. The pathogenesis of most CNS disorders is the result of the interplay of genetic and epigenetic factors with environmental factors leading to post-transcriptional changes and proteomic and metabolomic dysfunctions. It is estimated that genetics accounts for 20% to 95% of variability in drug disposition and pharmacodynamics, and about 25-60% of the Western population is defective in genes responsible for drug metabolism. In the European population only 25% of subjects are pure extensive metabolizers for the trigenic cluster integrated by the CYP2D6, CYP2C19 and CYP2C9 genes. About 50% of adverse drug events in CNS disorders might be attributed to pharmacogenomic factors. The rationale for practical pharmacogenomics and personalized therapeutics based on individual genomic profiles implies the management of different types of genes and their products including (i) genes associated with the mechanism of action of psychotropic drugs (neurotransmitters, receptors, transporters), (ii) genes encoding enzymes responsible for drug metabolism (phase I, phase II reactions), (iii) disease-specific genes associated with a particular pathogenic cascade), and (iv) pleiotropic genes with multilocative effects in metabolomic networks. The incorporation of genomic medicine procedures and pharmacogenomics into clinical practice, together with educational programs for the correct use of medication, must help to optimize therapeutics in CNS disorders.
[Back to top]


Non-alcoholic steatohepatitis: new insights from OMICS studies
Martel Cécile, Degli Esposti Davide, Bouchet Antonin, Brenner Catherine and Lemoine Antoinette BRENNER Catherine
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00313]

Non-alcoholic fatty liver disease (NAFLD) is the most common liver pathology characterized by fat accumulation in a context of metabolic syndrome or insulin resistance. It can be associated with obesity, diabetes, hyperinsulinemia, dyslipidemia as well as hypertension. NAFLD consists of a large spectrum of hepatic lesions including benign steatosis, non-alcoholic steatohepatitis (NASH), cirrhosis or hepatocellular carcinoma. Upon chronic stress, NASH would occur via at least “two-hits” process involving modulation of a high number of genes and proteins. Firstly, the accumulation of fat, either due to the increased inflow of free fatty acids or de novo lipogenesis, leads to steatosis. Secondly, when adaptive mechanisms for stress tolerance are overwhelmed, lipotoxicity and chronic inflammation trigger major hepatic damages, mainly via oxidative and inflammatory stress, lipid peroxidation and cell death. As a consequence, all these processes concur to favor steatohepatitis, fibrosis and cancer. Recently, the elucidation of physiopathological signaling cascades controlling NAFLD and NASH benefited from large-scale studies, namely the omics, such as transcriptomics, genomics, proteomics, and lipidomics. The advent of lipidomics would allow shedding light upon the respective roles of triglyceride and fatty acid metabolites in the lipotoxic liver injury hypothesis for the pathogenesis of NASH. In this review, the contribution of the omics to the understanding of the molecular basis of NASH is discussed that could offer perspectives for novel biomarkers discovery.
[Back to top]


Application of lipidomics to assess lipogenesis in drug development and pre-clinical trials
Antonin Lamazière, Claude Wolf and Peter J. Quinn
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00314]

The rising incidence of cardiovascular and metabolic diseases in industrialized countries has led the pharmaceutical industry to make them key areas of drug development. These diseases imply a clustering of metabolic factors where lipid metabolites play a relevant role. Measurement of pharmacodynamic endpoints of drugs on lipid metabolism pathways and downstream biological processes appear crucial for a rational drug discovery/development. Fortunately, recent mass spectrometers with an enhanced sensitivity and resolution in combination with multivariate statistical analysis provide the practical possibility to analyze and measure wide portions of the lipidome. The final goal is to identify lipid signatures which fit with specific pharmacologic responses to therapeutic intervention. Focusing on applications of lipidomics for drug development this review outlines the methodological steps, from analytical measurements to data processing and to graphical representation, for an efficient implementation of informative lipid signatures.
[Back to top]


Intestinal MicrobiOMICS to Define Health and Disease in Human and Mice
Matteo Serino, Chantal Chabo and Remy Burcelin
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00315]

Over the last five years an increasing effort has been made to understand the role of intestinal microbiota in health and disease, resulting in regarding to it as a new organ actively involved in the control of host metabolism, both in humans and mice. Amongst hundreds (up to thousand) germ species inhabiting the intestine, few of them are cultivable. Nevertheless, next-generation sequencing-based molecular technologies have been developed, allowing to overcome this problem and shed light on the way the gut microbiota undergoes dramatic changes during (patho)-physiological modifications of the host. Hence, the study of the overall gut germ genome (metagenome) and transcriptome (microbiome) has been launched. Thus, Genomics and Transcriptomics have begun to be increasingly used, opening the so called “Omics” era, including Proteomics and Metabolomics techniques as well. Taken together, the “Omics” allow the study of gut microbiota impact on whole host metabolism, resulting in the definition of new metabolic profiles (i.e. the presence of metabolites within the blood defines a metabolomic profile), others than those based on nucleic acid analyses only. Once demonstrated the involvement of gut microbiota within metabolic diseases, “Omics” analyses has allowed the identification of the obesity-induced gut microbiota imbalance, characterized by increased Firmicutes to Bacteroidetes ratio (metagenomics) and of the so called “core microbiome”, focusing on the gut microbiota at a gene- rather than, solely, at a taxonomic-level. In addition, metabolomics studies revealed, for instance, the implication of gut microbiota to nonalcoholic fatty liver disease in insulin-resistant mice. Additionally, the use of germ-free (axenic) mice has made possible the microflora transfer to investigate the mechanisms through which gut microbes modulate host metabolism, albeit the molecular actors of the host–gut-microbiota interplay remain to be fully determined. Here, we report the role of “Omics” in the multiple analyses of gut microbiota-driven metabolic modifications of the host, proposing also to focus on lipopolysaccharides (LPS), the Gram negative proinflammatory molecules we already showed to be the initiators of metabolic diseases.
[Back to top]


The glutamate hypothesis of schizophrenia: Neuroimaging and drug development
Alice Egerton and James M. Stone
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00317]

Over the last 50 years, evidence for central involvement of glutamatergic neurotransmission in the pathophysiology of schizophrenia has accumulated. Recent advances in neuroimaging technology now allow several components of glutamatergic neurotransmission to be assessed in the living human brain. Positron emission tomography (PET) or single photon emission tomography (SPET) in combination with select radiotracers allows visualization of glutamatergic receptors in vivo, and magnetic resonance (MR) – based techniques allow mapping of the effects of glutamatergic agents on regional brain activation, and the measurement of regional glutamate concentrations. These imaging studies have provided evidence for regional glutamatergic abnormalities in psychosis, and are beginning to describe both the evolution of these abnormalities over the course of the illness and their response to therapeutic intervention. In parallel, advances in small animal imaging and the development of animal models have provided a platform to explore the neuropathological consequences of glutamatergic abnormality, and the potential antipsychotic efficacy of novel compounds. The molecular diversity of the glutamatergic system has driven the design of several compounds targeting aspects of glutamatergic transmission, and clinical trials have yielded encouraging results. Here, we review the contribution of imaging studies to date in understanding glutamatergic abnormalities in psychosis, and discuss the potential of new glutamatergic compounds for treatment of the disorder.
[Back to top]


Selective activation of muscarinic acetylcholine receptors for the treatment of schizophrenia
Brian Dean
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00318]
It is being increasingly recognised that the future of drug development will need to be based on a comprehensive understanding of disease pathophysiology.  Thus this review focuses on a growing body of information suggesting that decreases in muscarinic receptors are involved in the pathophysiology of schizophrenia.  This review will address evidence to support the hypothesis that drugs that can increase the activity of muscarinic receptors have the potential to have antipsychotic affects and improve cognitive deficits in subjects with the schizophrenia.  How drugs directed towards allosteric binding sites are overcoming the problem of using orthosteric receptor agonists in the treatment of disease, because of their propensity to cause agonist-induced receptor down-regulation and subsequent drug desensitisation, will be discussed.  The discovery of allosteric binding sites on muscarinic receptors will be reviewed as will the ability of different classes of drugs to stimulate each muscarinic receptor without causing agonist-induced receptor down-regulation.  Finally, progress in developing allosteric muscarinic receptor agonists and modulators will be discussed and it will be argued the muscarinic M1 receptor allosteric agonists and/or modulators may have the potential to improve the cognitive deficits associated with schizophrenia whilst muscarinic M4 receptor modulates may have antipsychotic effects.
[Back to top]


Current progress in the genetic research of schizophrenia: relevance for drug discovery?
Dan Rujescu, Just Genius, Jens Benninghoff and Ina Giegling
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00319]

Schizophrenia is a devastating brain disease. The mode of inheritance is complex and non-Mendelian with a high heritability of ca. 65-80%. Given this complexity, until most recently it was notoriously difficult to identify disease genes. Due to new technologies the last few years have brought an explosion of interest in human genetics of complex diseases. The knowledge resulting from the availability of the complete sequence of the human genome, the systematic identification of single nucleotide polymorphisms (SNPs) throughout the genome, and the development of parallel genotyping technology (microarrays) established the conditions that brought about the current revolution in our ability to probe the genome for identifying disease genes. All these studies have opened a window into the biology of common complex diseases and have provided proof of principle and yielded a multitude of genes showing strong association with complex diseases. New findings in schizophrenia will be summarized in this review and discussed in the light of a possible translation into the development of better treatment.
[Back to top]


Serotonin Receptors as Targets for Drugs Useful to Treat Psychosis  and Cognitive Impairment in Schizophrenia
H. Y. Meltzer, B. W. Massey and M. Horiguchi
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00320]

The concept that the efficacy of all antipsychotic drugs (APDs) can be explained by their action on dopamine (DA) D₂ receptors is most challenged by drugs such as clozapine which target serotonin (5-HT)_2A receptors as an essential component of their efficacy and tolerability. The 5-HT_2A receptor, along with 5-HT_1A, 5-HT _2C, 5-HT ₆ or 5-HT ₇ receptors, all of which are components of the mechanism of action of clozapine, represent important targets for treating multiple aspects of schizophrenia, especially psychosis and cognitive impairment. The class of atypical antipsychotic drugs (APDs), of which clozapine is the prototype, share in common more effective 5-HT _2A receptor inverse agonism and weaker interference with D₂ receptor stimulation, either through D₂ receptor blockade or partial D₂ receptor agonism. This has led to development of a selective 5-HT_2A antagonist, ACP-103 (pimavanserin), which has been found to be effective as monotherapy in L-DOPA psychosis and has promise as an add-on agent for sub-effective doses of atypical APDs. We review here the extensive preclinical evidence to support the importance of 5-HT_2A receptor inverse agonism to the action of clozapine and related atypical APDs, and evidence supporting the potential of selective 5-HT_2A, 5-HT ₆ ,  and 5-HT ₇, antagonists, 5-HT_1A partial agonists and 5-HT_2C agonists for development of drugs which ameliorate psychosis or cognitive impairment.
[Back to top]


Treatment of Cognitive Dysfunction in Schizophrenia
H.M. Ibrahim  and C.A. Tamminga
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00321]

Cognitive impairment is a core feature of schizophrenia that substantially accounts for poor functional outcomes associated with this disease in areas such as work, independent living and social relationships. Until recently, drug development in schizophrenia has focused on developing compounds that mainly target the positive psychotic symptoms of the illness. Although current antipsychotic drugs treat psychosis in schizophrenia rather well, their impact on cognitive dysfunction is minimal. In recent years there has been growing interest in developing novel treatments for cognitive deficits in schizophrenia. In this review we discuss pharmacologic strategies considered most likely to improve cognition. These putative molecular targets include receptors for acetylcholine, dopamine, glutamate, g-aminobutyric acid (GABA), serotonin and histamine. In addition, we propose that not only pharmacological, but also psychological treatments should be considered to enhance cognition in schizophrenia.
[Back to top]


Disturbed function of GABAergic interneurons in schizophrenia: Relevance for medical treatment?
J. Genius, I. Giegling, J. Benninghoff and D. Rujescu
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00322]

For decades treatment of schizophrenia was restricted to drugs, which mainly target positive symptoms by interfering with the dopaminergic neurotransmission.

Since a large body of experimental and clinical data implicate that schizophrenia may primarily be a consequence of an imbalance in the glutamatergic system, specifically the networks containing GABAergic interneurons (γ-amino butyric acid), new drugs modulating glutamatergic neurotransmission are being developed. Targeting this dysfunction may follow different strategies, including application of direct or indirect NMDA (N-methyl-D-aspartate) receptor agonists or drugs modulating the function of metabotropic glutamate receptors. Meanwhile, the first substances have proven to be effective in animal models of schizophrenia and now enter the stage of clinical trials. The most promising data have been obtained in studies employing agonists of the metabotropic glutamate receptor. A choice of these substances is presented in this review.
[Back to top]


Metabotropic glutamate receptors as targets for novel antipsychotic treatments
L.J. Gray, A.J. Hannan and X. Zhang
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00323]

In recent years metabotropic glutamate receptors have emerged as key targets for the design of new antipsychotic medications for schizophrenia, in particular mGluR5 and mGluR2/3. These receptors exhibit diverse interactions with other neurotransmitter receptors and critical elements of intracellular signalling cascades known to be important to the pharmacotherapy of schizophrenia. In addition, mGluR5 and mGluR2/3 are intimately involved in behavioural domains related to the symptoms of this disorder. Both animal and clinical studies using novel drugs targeting these receptors have provided encouraging results. The number of patents registered for drugs targeting metabotropic glutamate receptors has grown dramatically, and positive allosteric modulators for both receptors show particular promise.
[Back to top]


Subjective well-being of patient with schizophrenia as a target of drug treatment
E. Karamatskos, C. Mulert, M. Lambert and D. Naber
[Purchase Article] [BSP/CPB/E-Pub/00324]

An important development within the last decades is the consideration of the patient's perspective and the acknowledgement that the majority of patients are able to judge their state of well-being. Several self-report scales such as the “The Subjective Well-being under Neuroleptics Scale” (SWN) have been established. Additionally to their beneficial impact, current antipsychotics have considerable limitations. Antipsychotic-related side effects, such as extrapyramidal-motor symptoms, weight gain and obesity, apathy and anhedonia have an important influence on the patient’s wellbeing. Evidence suggests that the so-called neuroleptic-induced deficit syndrome under antipsychotics might be caused by the inhibition of the dopaminergic reward system. A reduced activation of the ventral striatum, including the nucleus accumbens is associated with negative symptom severity. Second-generation antipsychotics (henceforth SGA) block striatal D2 receptors less and show a weaker binding to D2 receptors, have interactions with several other neurotransmitters and inhibit to a lesser degree the reward functions compared to first-generation antipsychotics (henceforth FGA). This may support the reduction of negative symptoms, contributes to a higher subjective well-being, a better medication adherence and thereby an improved therapeutic outcome. The involvement of the patient and the consideration of his/her subjective wellbeing will be a major aspect in the development of new treatment strategies in schizophrenia and has a significant impact on the adherence and the long-term prognosis.
[Back to top]


Immunological Treatment Options for Schizophrenia
Norbert Müller and Markus J. Schwarz
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00325]

The exact pathophysiological mechanism leading to dopaminergic dysfunction in schizophrenia is still unclear, but inflammation is postulated to be a key player: a dysfunction in the activation of the type 1 immune response seems to be associated with decreased activity of the key enzyme in tryptophan/kynurenine metabolism, indoleamine 2,3-dioxygenase (IDO), resulting in increased production of kynurenic acid - a N-methyl-D-aspartate (NMDA) antagonist in the central nervous system (CNS) - and reduced glutamatergic neurotransmission. The differential activation of microglia cells and astrocytes as functional carriers in the immune system in the CNS may also contribute to this imbalance. Existing antipsychotics, which predominantly act as dopamine D2 receptor antagonists, have several shortcomings. The immunological effects of many existing antipsychotics, however, rebalance in part the immune imbalance and the overproduction of kynurenic acid. The immunological imbalance results in an inflammatory state with increased prostaglandin E2 (PGE2) production and increased cyclo-oxygenase-2 (COX-2) expression. Growing evidence from clinical studies with COX-2 inhibitors points to favourable effects of anti-inflammatory therapy in schizophrenia, in particular in an early stage of the disorder. Further options for immunomodulating therapies in schizophrenia will be discussed.
[Back to top]


From Glutamatergic dysfunction to cognitive impairment: Boundaries in the therapeutic of the schizophrenia
Gaspar PA, Bustamante ML, Rojo LE and Martinez A
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00326]

Cognitive deficits are trait markers in schizophrenia and the improvement of these dysfunctions has been considered as a new frontier of treatment in this disease. A current model for the patophysiology of schizophrenia states that N-methyl-D-aspartate receptor (NMDA) hypofunction leads to a dysregulation of gamma-amino butyric acid (GABA) fast- spiking interneurons, consequently disinhibiting pyramidal glutamatergic output and disturbing signal-to-noise ratio. In this way, the modulation of the glutamate activity might constitute a highly promising target for future therapeutic interventions of this disease. In the present review, we discuss key regulatory elements for glutamatergic neurotransmission and provide new insights into their potential role in developing pharmacological treatments. Also, we emphasize the role of certain chemical families as potential sources of new lead compounds with affinity for metabotropic glutamate receptors (mGluRs) with cognitive enhancing properties.
[Back to top]


Pharmacotherapy of schizophrenic patients: Achievements, unsolved needs, future research necessities
Hans-Jürgen Möller
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00327]

Schizophrenia is a severe mental disorder, characterized by a complex symptomatology and a chronic relapsing course, leading to a more or less intensive residual syndrome. The disorder has strong consequences on social functioning and on the patients’ quality of life and is associated with a huge economical burden for the society. The best possible drug treatment should be therefore provided.

The introduction of the FGAs was an important step forward, since they were the first specific drug treatment for schizophrenia in the first place. However, the positive therapeutic effect was counterbalanced by problematic side-effects, predominantly different kinds of EPS.

The SGAs have a much lower liability to induce EPS, with clozapine inducing no EPS. Additionally, SGA demonstrated some advantages in terms of a broader spectrum of efficacy, including e.g. negative symptoms. However, for most of the single compounds this efficacy advantage was not as high as initially expected.

Thus, there is a need for new developments. Currently, drugs in development are based on the classical transmitter related approaches, following especially the model of clozapine and several other multi-receptor compounds.

More and more, the glutamatergic system is of interest in this context as well. Totally new directions, e.g. based on genetic findings or focussing more on brain plasticity are headed for. The question is, whether beside the classical concept of antipsychotics, which cover all symptomatic domains of schizophrenia, compounds demonstrating efficacy only in one syndrome such as e.g. negative symptoms will have a chance in the future development.
[Back to top]


Recent advances in targeting the ionotropic glutamate receptors in treating schizophrenia
Robert E. McCullumsmith, John Hammond, Adam Funk and James H. Meador-Woodruff
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00328]

The treatment of schizophrenia has been focused on modulation of dopamine receptors for over 50 years. Recent developments have implicated other neurotransmitter systems in the pathophysiology of this illness. The discovery and characterization of glutamate receptors and their roles in the brain has lead to novel approaches for the treatment of schizophrenia. In this article, we review drugs that modulate ionotropic gluamate receptors and discuss their efficacy for the treatment of this often debilitating severe mental illness.
[Back to top]


Functional Magnetic Resonance Imaging and treatment strategies in schizophrenia
Susanne Karch, Oliver Pogarell and Christoph Mulert
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/0029]

Neurobiological correlates of various treatment approaches (e.g. psychopharmacology, cognitive behavioural therapy) have become an important issue in recent neuroimaging studies. In schizophrenia, dysfunctions especially in frontal and parietal brain regions as well as the ventral striatum have been demonstrated in numerous studies. The review includes functional MRI studies about neurobiological correlates of treatment effects in schizophrenic patients. Treatment-associated changes in brain activity were shown especially in the anterior cingulate cortex, the dorsolateral prefrontal cortex, the ventral striatum and the amygdala. In addition, the differential effects of various treatment approaches including different neuroleptics (atypical neuroleptics, typical neuroleptics) as well as cognitive behavioural therapy will be considered in this review. The influence of functional differences on the differential effect of typical and atypical antipsychotics on cognitive functions will be discussed.
[Back to top]


DTNBP1 (Dysbindin) Gene Variants: In vivo evidence for effects on hippocampal glutamate status
C Wirth, F Schubert, M Lautenschlager, R Brühl, A Klär, T Majic, UE Lang, A Ehrlich, G Winterer, T Sander, M Schouler-Ocak and J Gallinat
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00330]

In linkage and association studies the DTNBP1 gene has been identified as a major susceptibility gene for schizophrenia. Reduced expression of DTNBP1 was found in the hippocampus and prefrontal cortex in post mortem brains of schizophrenic patients. In vitro and animal models provide evidence that the DTNBP1 gene product dysbindin modulates the activity of the neurotransmitter glutamate in hippocampal neurons and is crucial for cell functioning and synaptogenesis. This study is the first to investigate the effects of genetic variants of DTNBP1 on the status of the glutamate system as well as neuronal integrity (N-acetylaspartate, NAA) in the hippocampus and a cortical region, the anterior cingulate cortex (ACC), in humans.

Methods: In 79 healthy subjects, the association of single nucleotide polymorphisms (SNPs) rs760665 and rs909706 with absolute concentrations of glutamate and NAA in the left hippocampus and the ACC were investigated, using proton magnetic resonance spectroscopy (MRS) at 3 Tesla and a well established quantification procedure.

Results: Hippocampal glutamate concentration was significantly affected by genotype of rs760665 (F=4.406,df=2,p=0.016) and rs909706 (F=3.171,df=2,p=0.048). For the concentration of NAA, a weak association with rs760665 was observed in the contrast analysis. None of the metabolites measured in the ACC showed a significant connection with either genotype.

Conclusion:The results support a role of DTNBP1 gene variants in the glutamate neurotransmission system in the human brain at least in the hippocampus. This is compatible to growing evidence of a crucial role of glutamate in the pathobiology of schizophrenia. In addition, the weak association between DTNBP1 genotype and NAA is in line with a regulatory influence of dysbindin on synaptogenesis and neuronal survival.
[Back to top]


Innovative treatment approaches in schizophrenia enhancing neuroplasticity: Aerobic exercise, erythropoetin and repetitive transcranial magnetic stimulation
Thomas Wobrock,. Alkomiet Hasan and Peter Falkai
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00331]

Schizophrenia is a brain disorder associated with subtle, but replicable cerebral volume loss mostly prevalent in frontal and temporal brain regions. Post-mortem studies of the hippocampus point to a reduction of the neuropil constituting mainly of synapses associated with changes of molecules mediating plastic responses of neurons during development and learning. Derived from animal studies interventions to enhance neuroplasticity by inducing adult neurogenesis, synaptogenesis, angiogenesis and long-term potentiation (LTP) were developed and the results translated into clinical studies in schizophrenia. Out of these interventions aerobic exercise has been shown to increase hippocampal volume, elevate N-acetyl-aspartate in the hippocampus as neuronal marker, and improve short-term memory in schizophrenia. The hematopoietic growth factor erythropoetin (EPO) is involved in brain development and associated with the production and differentiation of neuronal precursor cells. A first study demonstrated a positive effect of EPO application on cognition in schizophrenia patients. In randomised controlled studies with small sample size, the efficacy of repetitive transcranial magnetic stimulation (rTMS), a biological intervention focussing on the enhancement of LTP, has been shown for the improvement of positive and negative symptoms in schizophrenia,. The putative underlying neurobiological mechanisms of these interventions including the role of neurotrophic factors are outlined and implications for future research regarding neuroprotection strategies to improve schizophrenia are discussed.
[Back to top]


Altered Cortical GABA Neurotransmission in Schizophrenia: Insights into Novel Therapeutic Strategies
Ana Stan and David A. Lewis
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00332]

Altered markers of cortical GABA neurotransmission are among the most consistently observed abnormalities in postmortem studies of schizophrenia.  The altered markers are particularly evident between the chandelier class of GABA neurons and their synaptic targets, the axon initial segment (AIS) of pyramidal neurons.  For example, in the dorsolateral prefrontal cortex of subjects with schizophrenia immunoreactivity for the GABA membrane transporter is decreased in presynaptic chandelier neuron axon terminals, whereas immunoreactivity for the GABA_A receptor α2 subunit is increased in postsynaptic AIS.  Both of these molecular changes appear to be compensatory responses to a presynaptic deficit in GABA synthesis, and thus could represent targets for novel therapeutic strategies intended to augment the brain’s own compensatory mechanisms. Recent findings that GABA inputs from neocortical chandelier neurons can be powerfully excitatory provide new ideas about the role of these neurons in the pathophysiology of cortical dysfunction in schizophrenia, and consequently in the design of pharmacological interventions.
[Back to top]


Investigation of Superparamagnetic Iron Oxide Nanoparticles for MR-Visualization of Surgical Implants
Slabu, G. Güntherodt, T. Schmitz-Rode, M. Hodenius, N. Krämer, H. Donker, G.A. Krombach, J.Otto, U. Klinge and M. Baumann
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00334]

For the development of a surgical mesh implant that is visible in magnetic resonance imaging (MRI), superparamagnetic iron oxides (SPIOs) are integrated into the material of the mesh. In order to get a high quality mesh regarding both mechanical and imaging properties, a narrow size distribution and homogenous spatial distribution, as well as a strong magnetization of SPIOs within the filament of the mesh are required. In this work, six different samples of SPIOs composed of a magnetite core are synthesized with and without stabilizing dodecanoic acid and analyzed using a superconducting quantum interference device (SQUID), transmission electron microscope (TEM) and a magnetic force microscope (MFM) to determine the properties that are beneficial for the assembly and imaging of the implant. These analyses show the feasibility of visualization of surgical implants with incorporated SPIOs and the influence of the agglomeration of SPIOs on their magnetization and on a homogenous spatial distribution within the polymer of the mesh.
[Back to top]


Monitoring Molecular, Functional and Morphologic Aspects of Bone Metastases Using Non-Invasive Imaging
Tobias Bäuerle, Dorde Komljenovic and Wolfhard Semmler
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00335]

Bone is among the most common locations of metastasis and therefore represents an important clinical target for diagnostic follow-up in cancer patients. In the pathogenesis of bone metastases, disseminated tumor cells proliferating in bone interact with the local microenvironment stimulating or inhibiting osteoclast and osteoblast activity. Non-invasive imaging methods monitor molecular, functional and morphologic changes in both compartments of these skeletal lesions – the bone and the soft tissue tumor compartment. In the bone compartment, morphologic information on skeletal destruction is assessed by computed tomography (CT) and radiography. Pathogenic processes of osteoclast and osteoblast activity, however, can be imaged using optical imaging, positron emission tomography (PET), single photon emission CT (SPECT) and skeletal scintigraphy. Accordingly, conventional magnetic resonance imaging (MRI) and CT as well as diffusion-weighted MRI and optical imaging are used to assess morphologic aspects on the macroscopic and cellular level of the soft tissue tumor compartment. Imaging methods such as PET, MR spectroscopy, dynamic contrast-enhanced techniques and vessel size imaging further elucidate on pathogenic processes in this  compartment including information on metabolism and vascularization. By monitoring these aspects in bone lesions, new insights in the pathogenesis of skeletal metastases can be gained. In translation to the clinical situation, these novel methods for the monitoring of bone metastases might be applied in patients to improve follow-up of these lesions, in particular after therapeutic intervention. This review summarizes established and experimental imaging techniques for the monitoring of tumor and bone cell activity including molecular, functional and morphological aspects in bone metastases.
[Back to top]


Imaging of hypoxia using PET and MRI
Gaertner FC, Souvatzoglou M, Brix G and Beer AJ
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00336]

Tumor hypoxia is the result of an inadequate supply of oxygen to tumor cells which can be caused by multiple factors. It is associated with aggressive local tumor growth and invasion, increased risk of metastasis, higher resistance to radiotherapy (RT) and chemotherapy, overall resulting in a poor clinical prognosis. Many locally advanced solid tumors may exhibit hypoxic and/or anoxic tissue areas that are heterogeneously distributed within the tumor mass.

As hypoxia is a negative prognostic factor concerning response to radiotherapy and chemotherapy, in vivo measurement of tumor hypoxia could be helpful to identify patients with worse prognosis or patients that could benefit from appropriate treatments such as intensity modulated radiotherapy (IMRT) that may accurately conform the dose distribution to small intratumoral regions showing differences in the oxygen level. A manifold of different methods to assess the oxygen tension (pO2) in tissues have been developed, each of them offering advantages as well as drawbacks. They range from invasive direct measurement techniques of the pO2 in tissue by using a polarographic electrode, to non-invasive imaging techniques such as positron emission tomography (PET) or magnetic resonance imaging (MRI). This article provides an overview over the various methods, with a particular emphasis on PET and MRI for imaging of hypoxia, and reviews their performance in preclinical and clinical studies.
[Back to top]


Cellular and Subcellular Imaging in Live Mice Using Fluorescent Proteins
Robert M. Hoffman
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00337]

Fluorescent proteins have revolutionized in vivo biology.  Due to their intrinsic brightness, multiple colors, and ease of generic manipulation, fluorescent proteins are the genetic reporters of choice for in vivo imaging.  The present report reviews applications of fluorescent protein for imaging cancer progression, gene expression, angiogenesis, stem cells, bacterial infection, Leishmania, and asthma at the cellular level in live mice.  With fluorescent protein-expressing cells and a highly sensitive small animal imaging system, cellular and subcellular dynamics can now be observed in live mice in real time. Fluorescent proteins thus enable both micro as well as macro imaging technology and thereby provide the basis for the new field of in vivo cell biology.
[Back to top]


Theranostic systems and strategies for monitoring nanomedicine-mediated drug targeting
Sijumon Kunjachan, Jabadurai Jayapaul, Marianne E. Mertens, Gert Storm, Fabian Kiessling and Twan Lammers
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00338]"

Nanomedicine formulations are considered to be superior to standard low-molecular-weight drugs because of an increased drug accumulation at the pathological site and a decreased localization to healthy non-target tissues, together leading to an improved balance between the efficacy and the toxicity of (chemo-) therapeutic interventions. To better understand and further improve nanomedicine-mediated drug targeting, it is important to design systems and strategies which are able to provide real-time feedback on the localization, the release and the therapeutic efficacy of these formulations. The advances made over the past few years with regard to the development of novel imaging agents and techniques have provided a broad basis for the design of theranostic nanomedicine materials, i.e. multicomponent carrier constructs in which drugs and imaging agents are combined, and which can be used to address issues related to drug localization, drug release and drug efficacy. Here, we summarize several recent efforts in this regard, and we show that theranostic systems and strategies hold significant potential for monitoring and improving nanomedicine-mediated drug targeting.
[Back to top]


Imaging in the age of molecular medicine: Monitoring of anti-angiogenic treatments
W. Lederle, M. Palmowski and F. Kiessling
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00339]

Angiogenesis is a complex multistep process and a crucial pre-requisite for tumor growth, invasion and metastasis. A profound knowledge of the mechanisms including the elucidation of markers for angiogenic vessels is essential for the generation of new anti-angiogenic chemotherapeutic agents and the improvement of specific imaging techniques. During the last decades, numerous angiogenesis inhibitors have been developed and some of them have shown promising results in preclinical and clinical trials. However, the response to anti-angiogenic treatment is often delayed and shows high inter-individual variations. In order to improve anti-angiogenic therapy, new specific surrogate markers are necessary that allow the characterization of different angiogenic steps, especially at the early stage. In this respect, non-invasive imaging is a potent tool for characterizing the tumor vascularization and for sensitive and longitudinal treatment monitoring. In particular, new molecular imaging techniques might ultimately improve the characterization of the angiogenic tumor phenotype and stage. This review summarizes the current status of different imaging modalities e.g. MRI, CT, US, nuclear and optical imaging with respect to the imaging of tumor angiogenesis and of anti-angiogenic treatments. It also includes new approaches in molecular imaging, which give deep insight into the tumor stage and the response of tumor vessels to anti-angiogenic therapy. Thus, this may lead to a more personalized cancer therapy in future.
[Back to top]


Translational Optical Imaging in Diagnosis and Treatment of Cancer
Stijn Keereweer, Merlijn Hutteman, Jeroen D.F. Kerrebijn, Cornelis J.H. van de Velde, Alexander L. Vahrmeijer and Clemens W.G.M. Löwik
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00340]

In cancer imaging, many different modalities are used that each have their specific features, leading to the combined use of different techniques for the detection, staging and treatment evaluation of cancer. Optical imaging using near-infrared fluorescence light is a new imaging modality that has recently emerged in the field of cancer imaging. After extensive preclinical research, the first steps of translation to the clinical practice are currently being made.

In this article, we discuss the preclinical and clinical results of near-infrared optical imaging for non-invasive detection and classification of tumors, therapy monitoring, sentinel lymph node procedures, and image-guided cancer surgery. Widespread availability of imaging systems and optical contrast agents will enable larger studies on their clinical benefit and can help establish a definitive role in clinical practice.
[Back to top]


Apoptosis imaging to monitor cancer therapy: The road to fast treatment evaluation?
Marijke De Saint-Hubert, Matthias Bauwens, Alfons Verbruggen and Felix M. Mottaghy
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00341]

Molecular imaging of biological processes may allow detection of therapy effects before the tumor is reduced in size. The most frequently used PET tracer in oncology, 2-[18F]fluoro-2-deoxyglucose (FDG), suffers from low specificity due to uptake in inflammatory cells. The proliferation marker, 3’-[18F]fluoro-3’-deoxy-L-thymidine (FLT), is less influenced by the inflammatory response following therapy but here disease- and drug-specific effects need to be considered.

Since cancer therapy mainly intends to eliminate cancer cells, imaging of cell death offers a direct way to image therapy response. This review gives an overview of the radiopharmaceutical development and in vivo evaluation of radioligands that have emerged so far for detection and assessment of apoptosis and necrosis.  Two radiopharmaceuticals that can image cell death have made it to clinical trials for follow up of tumor treatment: i) ^99mTc-and ¹²³I-labelled AnxA5  for the response to treatment of for example lymphoma and lung cancer and ii) ¹⁸F-ML10 for the evaluation of brain tumors post-radiation.  Other agents need further optimization.
[Back to top]


Deep Tissue Optical and Optoacoustic Molecular Imaging Technologies for Small Animal Research and Drug Discovery
Daniel Razansky, Nikos Deliolanis, Claudio Vinegoni and Vasilis Ntziachristos
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00342]

For centuries, biological discoveries were based on optical imaging, in particular microscopy but also several chromophoric assays and photographic approaches. With the recent emergence of methods appropriate for bio-marker in vivo staining, such as bioluminescence, fluorescent molecular probes and proteins, as well as nanoparticle-based targeted agents, significant attention has been shifted toward in vivo interrogations of different dynamic biological processes at the molecular level. This progress has been largely supported by the development of advanced tomographic imaging technologies suitable for obtaining volumetric visualization of bio-marker distributions in small animals at a whole-body or whole-organ scale, an imaging frontier that is not accessible by the existing tissue-sectioning microscopic techniques due to intensive light scattering beyond the depth of a few hundred microns. Major examples of such recently developed optical imaging modalities are reviewed here, including bioluminescence tomography (BLT), fluorescence molecular tomography (FMT), and optical projection tomography (OPT). The pharmaceutical imaging community has quickly appropriated itself of these novel forms of optical imaging, since they come with very compelling advantages, such as quantitative three-dimensional capabilities, direct correlation to the biological cultures, easiness and cost-effectiveness of use, and the use of safe non-ionizing radiation. Some multi-modality approaches, combining light with other imaging modalities such as X-Ray CT or MRI, giving the ability to acquire both an optical contrast reconstruction along with a hi-fidelity anatomical images, are also reviewed. A separate section is devoted to the hybrid imaging techniques based on the optoacoustic phenomenon, such as multispectral optoacoustic tomography (MSOT), which are poised to leverage the traditional contrast and specificity advantages of optical spectrum by delivering an ever powerful set of capabilities, including real-time operation and high spatial resolution, not affected by the scattering nature of biological tissues.
[Back to top]


Enzymatically Activatable Diagnostic Probes
Thorek DLJ and Grimm J
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00343]

Molecular imaging of disease development, progression and treatment is seen as key to further advancement in the understanding and triumph over illness. The role of enzymes is to catalyze the biochemical reactions that help regulate health, and when dysregulated in complex organisms lead to or indicate disease. The ability to image the action of these proteins for diagnostic purposes opens a window for the researcher and clinician to witness specifc molecular events in vitro and in vivo. Such probes have been developed and deployed for the optical, radionuclide and magnetic resonance modalities and offer significant benefits over conventional agents. The signal of enzymatically-activated probes is regulated by the specific activity of the desired enzyme. This allows for a higher signal to background ratio over non-specific and targeted agents. It also enables the modulation of contrast agent distribution (and even cellular accumulation) following enzymatic activity. This review summarizes the strategies and probes in development and use in this emergent field of molecular imaging, with a particular focus on the research and medical relevance of these advances.
[Back to top]


Carbon monoxide, a Two-Face for the protection of the liver
Florian Bösch and Tung Yu Tsui
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00345]

Carbon monoxide is generally believed to be a ‘toxic’ gas molecule due to its binding capability with hemoglobin. Overexposure to carbon monoxide leads to a hypoxic state that may cause the death of a mammalian. In contrast, directly exposure of carbon monoxide may protect cells or organs from various disease insults. The paradox effects of carbon monoxide might vary on the ways of exposure and the amounts being exposed. Here we highlighted the characteristics of this gas molecule and summarized its protective effects and therapeutic potentials in liver diseases and liver transplantation.
[Back to top]


Role of Carbon monoxide in vascular diseases
Ignazio Barbagallo, Giuseppina Marrazzo, Alessandro Frigiola, Agata Zappala and Giovanni Li Volti
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00346]

During the degradation of heme by the enzyme heme oxygenase (HO), Carbon monoxide (CO) is generated. Although it is considered as a non - significant and potentially toxic waste gas of heme catabolism, CO is a key signaling molecule used to regulate different cardiovascular functions. In this review, we focus the protective roles of CO in vascular injury/disease, which may be important to explore the overall protective roles of HO-1/CO system in the pathogenesis of human vascular disease. Key Words: Carbon monoxide, Heme oxygenase, oxidative stress, hypertension, vascular diseases.
[Back to top]


Role of Carbon Monoxide in Kidney Function: Is a little Carbon Monoxide Good for the Kidney?
Eva Csongradi, Luis A. Juncos, Heather A. Drummond, Trinity Vera and David E. Stec
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00347]

Carbon monoxide (CO) is an endogenously produced gas resulting from the degradation of heme by heme oxygense or from fatty acid oxidation. Heme oxygenase (HO) enzymes are constitutively expressed in the kidney (HO-2) and HO-1 is induced in the kidney in response to several physiological and pathological stimuli.  While the beneficial actions of HO in the kidney have been recognized for some time, the important role of CO in mediating these effects has not been fully examined. Recent studies using CO inhalation therapy and carbon monoxide releasing molecules (CORMs) are demonstrating that increases in CO alone can be beneficial to the kidney in several forms of acute renal injury by limiting oxidative injury, decreasing cell apoptosis, and promoting cell survival pathways.  Renal CO is also emerging as a major regulator of renal vascular and tubular function acting to protect the renal vasculature against excessive vasoconstriction and to promote natriuresis by limiting sodium reabsorption in tubule cells.  Within this review, recent studies on the physiological actions of CO in the kidney will be explored as well as the potential therapeutic avenues that are being developed targeting CO in the kidney which may be beneficial in diseases such as acute renal failure and hypertension.
[Back to top]


Heme Oxygenase-1/CO as Protective Mediators in Cigarette Smoke-Induced Lung Cell Injury and Chronic Obstructive Pulmonary Disease
Tamás Dolinay, Augustine M. K. Choi and Stefan W. Ryter
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00348]

Chronic obstructive pulmonary disease (COPD) is a disease involving airways restriction, alveolar destruction, and loss of lung function, primarily due to cigarette smoke (CS) exposure. The inducible stress protein heme oxygenase-1 (HO-1) has been implicated in cytoprotection against the toxic action of many xenobiotics, including CS. HO-1 also protects against elastase-induced emphysema. Differential expression of HO-1 in epithelial cells and macrophages may contribute to COPD susceptibility. Genetic polymorphisms in the HO-1 gene, which may account for variations in HO-1 expression among subpopulations, may be associated with COPD pathogenesis. Carbon monoxide (CO), a primary reaction product of HO-1 has been implicated in cytoprotection in many acute lung injury models, though it’s precise role in chronic CS-induced lung injury remains unclear. CO is a potential biomarker of CS exposure and of inflammatory lung conditions. To date, a single clinical trial has addressed the possible therapeutic potential of CO in COPD patients. The implications of the cytoprotective potential of HO-1/CO system in CS-induced lung injury and COPD are discussed.
[Back to top]


Carbon Monoxide In Acute Lung Injury
Simone Faller and Alexander Hoetzel
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00349]

Despite modern clinical practice in critical care medicine, acute lung injury still causes unacceptably high rates of morbidity and mortality. Therefore, the challenge today is to identify new and effective strategies in order to improve the outcome of these patients. Carbon monoxide, endogenously produced by the heme oxygenase enzyme system, has emerged as promising gaseous therapeutic that exerts protective effects against inflammation, oxidative and mechanical stress, and apoptosis, thus potentially limiting acute lung injury. In this review we discuss the effects of inhaled carbon monoxide on acute lung injury that results from ischemia-reperfusion, transplantation, sepsis, hyperoxia, or mechanical ventilation, the latter referred to as ventilator-induced lung injury. Multiple investigations using in vivo and in vitro models have demonstrated anti-inflammatory, anti-apoptotic, and anti-proliferative properties of carbon monoxide in the lung when applied at low dose prior to or during stressful stimuli. The molecular mechanisms that are modulated by carbon monoxide exposure are still not fully understood. Carbon monoxide mediated lung protection involves several signaling pathways including mitogen activated protein kinases, nuclear factor-κB, activator protein-1, Akt, peroxisome proliferating-activated receptor-γ, early growth response-1, caveolin-1, hypoxia-inducible factor-1α, caspases, Bcl-2-family members, heat shock proteins, or molecules of the fibrinolytic axis. At present, clinical trials on the efficacy and safety of CO investigate whether the promising laboratory findings might be translatable to humans.
[Back to top]


Cytoprotection by Inhaled Carbon Monoxide before Cardiopulmonary Bypass in Preclinical Models
Torsten Loop, Christian Schlensak and Ulrich Goebel
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00350]

Although a potentially toxic gaseous molecule, carbon monoxide recently gains rising scientifically and clinical interest as its beneficial effects and mechanisms of action are defined substantially in various in vitro and in vivo experiments. Its anti-inflammatory, anti-apoptotic and anti-proliferative properties but its increasing impact concerning numerous disease models in means of protection, well describe this gas as a new and challenging therapeutic alternative.

In this review, we focus on the extensively analyzed advantageous value of pre- and postconditioning with inhaled carbon monoxide in the context of lung and kidney injury, induced by the low perfusion during and after cardiopulmonary bypass. Mechanisms like the heat shock response as well as an expanded view regarding toxicity and side effects are described broadly.
[Back to top]


Application of Carbon Monoxide for Transplantation
Atsunori Nakao and Yoshiya Toyoda
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00351]

Carbon monoxide (CO) is an invisible, chemically inert, colorless and odorless gas and is toxic at high concentrations due to its interference with oxygen delivery. However, CO is endogenously and physiologically generated in mammalian cells via the catabolism of heme in a rate-limiting step of heme oxygenase systems, and CO potently protects against cellular injury. CO relaxes blood vessels and exerts anti-thrombotic effects by inhibiting platelet aggregation and derepressing fibrinolysis. In addition, CO reduces ischemia/reperfusion injury and inflammatory responses. CO inhibits apoptosis of endothelial and epithelial cells and reduces proliferation of smooth muscle cells, fibroblasts and T lymphocytes. Thus, there is accumulating evidence to support the notion that CO treatment of transplant donors, organs, or recipients can prevent graft dysfunction due to rejection or ischemia/reperfusion injury.  This invited review discusses recent advances and current knowledge pertaining to CO research in the field of transplantation.  In addition, we will discuss the clinical applicability of CO as a promising therapeutic strategy for the treatment of transplant patients.
[Back to top]


Carbon Monoxide – Toxicity of Low-Dose Application
Rene Schmidt, Helen Ryan and Alexander Hoetzel
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00352]

Carbon monoxide (CO) has long been considered a purely toxic by-product of incomplete combustion processes. Acute exposure to high concentrations of CO is one of the leading causes of fatal poisoning in industrial countries. However, after two decades of intensive research, there is ample evidence that CO endogenously produced by heme oxygenase enzymes has essential physiological functions and is of vital importance for cellular hemostasis. Furthermore, exogenously applied CO in low concentrations mediates potent cytoprotective effects. An overwhelming number of different in vitro and in vivo models demonstrated the protective action of CO application, e.g., in ischemia/reperfusion, transplantation, oxidative stress, inflammation, and others. Protection by this gaseous molecule could be illustrated for most organs, most species, and for different routes of administration. Now being on the verge of entering clinical trials, the question emerges whether the administration of low-dose CO would be safe for patients when used as a potential therapeutic. Therefore, this review summarizes in particular toxicological data obtained from low-dose CO exposure and discusses its impact on a possible clinical application.
[Back to top]


Carbon Monoxide and the Pancreas
Christian I. Schwer
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00353]

Carbon monoxide (CO), often referred to as the silent killer, is a colorless, odorless and tasteless gas. It combines with hemoglobin to produce carboxyhemoglobin, which is ineffective for delivering oxygen to animal and human tissues. On the other hand, CO is endogenously produced in the body as a byproduct of heme degradation catalyzed by the heme oxygenase (HO) enzymes. In the past decade, evidence has accumulated to suggest important physiological roles for CO in mammalian tissues. In the pancreas, modulation of endogenous CO production or administration of exogenous CO may represent a therapeutic option for the treatment of endocrine and exocrine pancreatic disorders. In cell culture, CO exerts anti-diabetic effects and brief exposure of purified mouse islets to CO ameliorates functional performance after transplantation. Recent advances include the observation that CO carriers possess potent anti-proliferative effects in an in vitro model of pancreatic fibrosis. In vivo, CO confers tissue protection in animal models of pancreatic disease, including those with hyperglycemia and inflammatory injury of the gland. However, there are still a number of unanswered questions surrounding its physiological and pathophysiological relevance and the preferred route of CO administration in the pancreas still remains to be settled. This brief review focuses on the roles, effects and mechanisms of action of CO in the pancreas.
[Back to top]


Antibacterial Effects of Carbon Monoxide
Jayne Louise Wilson, Helen E Jesse, Robert K Poole and Kelly S Davidge
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00354]

Carbon monoxide (CO) is a colourless and odourless gas that has long been considered as a potent respiratory poison. Recent advances have demonstrated its production by haem oxygenases in both mammals and microbes, and it has roles as a gasotransmitter in higher organisms. This review concentrates on the application of CO, via carbon monoxide-releasing molecules (CO-RMs), as an anti-bacterial agent. Currently, the scope of literature on the effects of CO on bacteria is small, and we have included discussions on the production of CO by bacteria via haem oxygenase enzymes, the use of CO as an energy source, and existing knowledge on CO sensors in bacteria. CO is known to target haem proteins and is an effective inhibitor of respiration, even when provided at concentrations much higher than prevailing oxygen. We review here data suggesting that CO-RMs are more effective inhibitors of respiration than is CO gas, perhaps due to the ability of CO-RMs to deliver CO selectively to intracellular targets. We also consider the recently reported transcriptomic consequences of CO-RM treatment of Escherichia coli, revealing a myriad of unexpected targets for CO and potential CO sensors. Finally, we consider the use of CO and CO-RMs as anti-bacterial agents in vivo, and the future prospects for this gaseous molecule.
[Back to top]


Reducing the development of antibiotic resistance in critical care units
Marjolein Deege and David L. Paterson
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00355]

Bacteria becoming resistant to an increasing number of antibiotic classes are a major problem at hospitals including critical care units worldwide. Awareness of this problem and the need to prevent the development of antibiotic resistance are very important, especially since very few new antibiotics will become available in the near future.

This article gives an overview of the mechanisms of antibacterial resistance and actual resistance data worldwide of the most prevalent Gram positive (MRSA, VISA/VRSE and VRE) and Gram negative bacteria (Pseudomonas aeruginosa, Acinetobacter spp., ESBL producing Enterobacteriaceae and Stenotrophomonas maltophilia). Furthermore, strategies to reduce antibiotic resistance are reviewed. Most important is institution of infection control policies including guidelines on surveillance, isolation of colonized patients and contact precautions, hand hygiene, decolonization measures and environmental decontamination. Antimicrobial stewardship, or striking the balance between an optimal antibiotic treatment for a patient and a minimal risk of development of antibiotic resistance, is another important strategy. Finally, optimizing of antibiotic dosage regimens and thus avoiding underdosage is essential to avoid selection of the most resistant subpopulation of bacteria during antibiotic treatment.

Intensive care units with knowledge of local epidemiology of resistance, an effective infection control program and antimicrobial stewardship policy tailored to their specific needs, and using optimal antibiotic dosing regimens have both locally decreased the risk of an outbreak with multi-resistant bacteria, and maybe even more important help to reduce the development of antibiotic resistance.
[Back to top]


A novel lily-of-the-valley fragrance contrast agent for magnetic resonance and fluorescence imaging of prostate cancer cells
Alexander Sturzu, Hartmut Echner, Uwe Klose, Sumbla Sheikh, Thomas Nägele, Christian Schwentner, Ulrike Ernemann and Stefan Heckl
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00356]

Detection of prostate carcinoma metastases is currently performed either via indirect tests like the prostate specific antigen (PSA) or prostate cancer gene 3 (PCA3) or by biopsies from masses found with medical imaging methods.

Our goal was to use an ectopic odorant receptor to target prostate-derived cells throughout the body for imaging by magnetic resonance and fluorescence imaging.

We synthesized a conjugate containing undecylic aldehyde (an antagonist of the human olfactory receptor hOR17-4), gadolinium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (a common magnetic resonance contrast agent) and fluorescein isothiocyanate (a fluorescent dye).

Two different prostate cancer cell lines as well as five other different malignant cell lines and healthy prostate epithelial cells were incubated with this conjugate and evaluated by flow cytometry, confocal laser scanning microscopy and magnetic resonance imaging.

The prostate-derived healthy and malignant cells showed stronger fluorescence than the non-prostate cancer cell lines in the flow cytometry and confocal laser scanning microscopy experiments.

In the magnetic resonance imaging experiments the T1 relaxation time reduction (higher signal intensity) was also stronger for the prostate-derived cells than for the non-prostate cells.

The examined conjugate showed high prostate-cell-specificity. This property makes it of potential value in the diagnosis of prostate cancer lymph node metastases.
[Back to top]


Pharmacokinetics/Pharmacodynamics Model-Supported Early Drug Development
Bin Chen, Jennifer Q. Dong, Wei-Jian Pan and Ana Ruiz
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00358]

Pharmacokinetic/pharmacodynamic (PK/PD) modeling & simulation (M&S) provides quantitative assessment of dose/exposure-response relationships with extensive applications at the late stage drug development as well as during regulatory decision making.  However, at preclinical and early phase clinical drug development, the importance of PK/PD M&S has not been as widely recognized.  We reviewed selected PK/PD M&S literatures in order to convey importance of M&S in these early development phases.  We focused on the application of M&S to select and optimize lead candidates, the use of preclinical PK/PD data to project the range of clinical doses, and the development of comprehensive dose/exposure-response models that can be used to forecast the probability of achieving a target response based on Phase 1 safety, PK and biomarker information.  We also reviewed several other M&S approaches that are often used in early drug development such as physiologically-based pharmacokinetic (PBPK) modeling, meta-analysis, PK-pharmacogenomics modeling, and etc. Our aims were to provide expert opinions on the practical utility of PK/PD model-based approaches that have positive impact on early decision-making with the goal of improving the success rate of early to late stage drug development.
[Back to top]


Ultrasound-Induced Blood-Brain Barrier Opening
Elisa E. Konofagou, Yao-Sheng Tung, James Choi, Thomas Deffieux, Babak Baseri and Fotios Vlachos
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00359]

Over 4 million U.S. men and women suffer from Alzheimer's disease; 1 million from Parkinson's disease; 350,000 from multiple sclerosis (MS); and 20,000 from amyotrophic lateral sclerosis (ALS). Worldwide, these four diseases account for more than 20 million patients. In addition, aging greatly increases the risk of neurodegenerative disease. Although great progress has been made in recent years toward understanding of these diseases, few effective treatments and no cures are currently available. This is mainly due to the impermeability of the blood-brain barrier (BBB) that allows only 5% of the 7000 small-molecule drugs available to treat only a tiny fraction of these diseases.  On the other hand, safe and localized opening of the BBB has been proven to present a significant challenge. Of the methods used for BBB disruption shown to be effective, Focused Ultrasound (FUS), in conjunction with microbubbles, is the only technique that can induce localized BBB opening noninvasively and regionally. FUS may thus have a huge impact in trans-BBB brain drug delivery. The primary objective in this paper is to elucidate the interactions between ultrasound, microbubbles and the local microenvironment during BBB opening with FUS, which are responsible for inducing the BBB disruption.  The mechanism of the BBB opening in vivo is monitored through the MRI and passive cavitation detection (PCD), and the safety of BBB disruption is assessed using H&E histology at distinct pressures, pulse lengths and microbubble diameters. It is hereby shown that the BBB can be disrupted safely and transiently under specific acoustic (pressures under 0.45 MPa) and microbubble (diameter under 8 μm) conditions.
[Back to top]


Experimental Methods and Transport Models for Drug Delivery across the Blood-Brain Barrier
Bingmei M Fu
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00360]

The blood-brain barrier (BBB) is a dynamic barrier essential for maintaining the micro-environment of the brain. Although the special anatomical features of the BBB determine its protective role for the central nervous system (CNS) from blood-born neurotoxins, however, the BBB extremely limits the therapeutic efficacy of drugs into the CNS, which greatly hinders the treatment of major brain diseases. This review summarized the unique structures of the BBB, described a variety of in vivo and in vitro experimental methods for determining the transport properties of the BBB, e.g., the permeability of the BBB to water, ions, and solutes including nutrients, therapeutic agents and drug carriers, and presented newly developed mathematical models which quantitatively correlate the anatomical structures of the BBB with its barrier functions. Finally, on the basis of the experimental observations and the quantitative models, several strategies for drug delivery through the BBB were proposed.
[Back to top]


Nanocarriers for the Simultaneous Co-Delivery of Therapeutic Genes and Anticancer Drugs
Nazia Choudhury and Huixin He
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00361]

Due to the molecular complexity of cancer, combination therapy is becoming increasingly important for better long-term prognosis with fewer side effects. To further increase the therapeutic effects, advanced drug delivery systems (DDSs), capable of simultaneously delivering multiple drugs to the site of action with specific time-programmed release profiles, are important requirements.  Nanocarriers for the simultaneous co-delivery of multiple chemical drugs in combination therapy have been extensively reviewed. Here we focus on the nanotechnology enabled DDSs for the simultaneous co-delivery of therapeutic genes and chemical drugs for cancer treatment. The opportunities for this combination strategy and their challenges will be discussed.
[Back to top]


Challenges and Strategies in Developing Microneedle Patches For Transdermal Delivery of Protein and Peptide Therapeutics
Fei Wu, Sixing Yang, Weien Yuan and Tuo Jin
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00362]

The birth of microneedles, an array of needles sufficiently long to penetrate epidermis but small enough to do not cause skin injury and pain feeling, has offered a highly promising solution for non-invasive delivery of protein and peptide drugs, a long-cherished desire over eighty years. However, the attempts to develop clinically feasible microneedle transdermal delivery methods encountered series of difficulties, for which a decade research efforts have yet to result in a single product. Microneedles may be incorporated into devices as skin pre-treatment tools, skin microinjectors as well as transdermal patches by their functions in drug delivery. They may also be categorized to insoluble solid microneedles, hollow microneedles, soluble/degradable solid microneedles and phase-transition microneedles by their structure and forming materials. This review article is aimed to update the progress and discuss the technical challenges raised in developing protein/peptide loaded microneedle patches.
[Back to top]


Release of Growth Factors, Cytokines and Therapeutic Molecules by Hyaluronan-based Hydrogels
Robert A. Peattie
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00363]

Hyaluronan (HA) is a highly biocompatible biopolymer that is widely used for a variety of therapeutic purposes including surgical preparations, adhesion prevention, viscosupplementation and drug and cytokine delivery.  Delivery can be accomplished effectively when HA-based carriers are synthesized in the form of hydrogels, though doing so normally requires chemical modification of the native HA structure.  Solute delivery from HA-based gels can be either “simple”, that is from a gel not including separate components intended to control release, or “regulated” when specific components are included for that purpose.  A variety of modified forms of HA have been developed and used for delivery of desired molecules in therapeutic, clinical, veterinary and laboratory research environments, and the number of such applications is likely to grow in future years.
[Back to top]


Nanocarriers to solid tumors: considerations on tumor penetration and exposure of tumor cells to therapeutic agents
Manali Bhagat, Susan Halligan and Stavroula Sofou
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00364]

Solid tumors constitute the majority of diagnosed cancers. For effective killing, therapeutic agents should ideally be delivered uniformly and at lethal doses to all cancer cells comprising the tumors, while keeping normal organ toxicities to a minimum. This requirement sets two of the major challenges in drug delivery to solid cancers: uniformity in delivery, and delivery of at least a minimum amount of therapeutics per cancer cell. Herein we review various approaches that aim to improve the penetration and content release of delivered therapeutic agents from nanocarriers of self-assembling nature. Biophysical characteristics of solid tumors are briefly discussed to motivate and rationalize the design of reported nanoparticle structures. This review does not aim to be exhaustive of the various designs and strategies, but to mostly give a flavor of the general current directions aiming to address these challenges.
[Back to top]


Critical Issues In Delivery of RNAi Therapeutics In Vivo
Stephanie Rivera and Fan Yuan
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00365]

RNA interference (RNAi) is a fundamental mechanism of gene regulation and has been harnessed to produce a new class of drugs for treatment of various diseases. A key issue in these applications is how to effectively deliver RNAi therapeutics into target cells. This review is focused on advances in RNA delivery in vivo. To achieve it, novel strategies have been developed to enhance stability of RNA in cells and tissues, overcome barriers to transport of RNA or its carriers in the body, and reduce immunogenicity and cytotoxicity of treatment. Approaches to RNA delivery are divided into three categories in this review: biological, chemical, and physical. Advantages and disadvantages of each method are discussed. At present, effective delivery of RNAi therapeutics in vivo is still a challenge although significant advances have been made in this field.
[Back to top]


Psoriasis, from pathogenesis to therapeutic strategies: IL-21 as a novel potential therapeutic target
Elisabetta Botti, Giulia Spallone, Roberta Caruso, Giovanni Monteleone, Sergio Chimenti and Antonio Costanzo
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00367]

Psoriasis is a common (1–3% of the population worldwide), multifactorial, immune-mediated chronic skin disease. In psoriasis pathogenesis an over-reaction of local innate immune response initiates inflammation with subsequent involvement of adaptive immune response leading to the production of a panel of cytokines, chemokines and growth factors leading to epidermal hyperplasia. Recently, IL-21 has been involved in this process as this cytokine is overexpressed in psoriatic skin and can cause  epidermal hyperplasia and inflammation when injected intradermally into mice. Moreover blockade of IL-21 with a human antibody against IL-21 reduces the epidermal thickness and the expression of Th1 and Th17 genes in the well-characterized model of human psoriasis-xenograft mouse. Therefore, the inhibition of this cytokine may be therapeutically effective in the treatment of psoriasis. Here we will review recent data on psoriasis pathogenesis focusing on the role of IL-21 as novel therapeutic target.
[Back to top]


Role of Defensins and Cathelicidin LL37 in Auto-immune and Auto-inflammatory diseases
Loredana Frasca and Roberto Lande
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00368]

Defensins and cathelicidins are anti-microbial peptides (AMPs) that act as natural antibiotics and are part of the innate immune defence in many species. We consider human defensins and LL37, the only human member of the cathelicidin family. In particular, we refer to the human alpha-defensins called human neutrophil peptides (HNP1 through 4), which are produced by neutrophils, HD5 and HD6, mainly expressed in Paneth cells of intestine, the human beta-defensins HBD1, HBD2 and HBD3, synthesized by epithelial cells and LL37, which is located in granulocytes, but is also produced by epithelial cells of the skin, lungs, and gut. In the last years, the study of AMPs activity and regulation has allowed to understand the important role of these peptides not only in the innate defence mechanisms against bacteria, viruses, fungi, but also in the regulation of immune cell activation and migration. Complementary studies have disclosed a role for AMPs in modulating many physiological processes that involve non-immune cells, such as activation of woud healing, angiogenesis, cartilage remodeling. Due to the pleiotropic tasks of these peptides, many of them are now being discovered to contribute to immune pathology of chronic diseases that affect skin, gut, joints; this is supported by many examples of immune-mediated pathologies in which their expression is disregulated. In this article we review the current literature that suggests a role for human defensins and LL37 in pathogenic mechanisms of several chronic diseases that are considered of auto-immune or auto-inflammatory origin.
[Back to top]


New pharmaceutical concepts for sebaceous gland diseases: implementing today’s pre-clinical data into tomorrow’s daily clinical practice
Vasiliki A. Zampeli, Evgenia Makrantonaki, Thrasivoulos Tzellos and Christos C. Zouboulis
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00369]

The human sebaceous gland is a microscopic branched type multiacinar gland been present everywhere on the body except on the palms and soles, whereas they are sparsely located on the dorsum of hands and feet. Several medical conditions are related with sebaceous gland pathology, such as acne, sebaceous hyperplasia, sebaceous adenoma and sebaceous carcinoma. Acne is a common, complex, chronic disorder of the human pilosebaceous unit that mostly occurs in adolescence and young adulthood. The sebaceous gland plays an exquisite role in the initiation of the disease. The multifactorial nature of the pathogenesis of acne includes increased sebum production, alteration of the quality of sebum lipids, inflammatory processes, interaction with neuropeptides and dysregulation of the hormone microenvironment, follicular hyperkeratinization and inflammation maintained by Propionbacterium acnes products within the follicle. On the other hand, the sebaceous gland, as a major and critical compartment of human skin, is also affected through ageing, both intrinsic and extrinsic, which lead to distinct clinical and histological changes. Intrinsic ageing of the sebaceous gland is determined primarily by genetic factors and hormonal status, with androgens playing a major role. A clinical manifestation associated with intrinsic ageing changes is skin xerosis. Extrinsic ageing of human sebaceous gland is mainly caused by accumulating UV irradiation, especially UVA. Photoageing of sebaceous gland is expressed with a wide spectrum of benign and malignant sebaceous tumours, such as sebaceous hyperplasia, sebaceous carcinoma and Muir-Torre syndrome. This review will focus on the pathogenesis of the most common sebaceous gland diseases and their molecular pathways which may represent future pharmaceutical targets.
[Back to top]


Gene Therapy of Skin Adhesion Disorders
Alessia Cavazza and Fulvio Mavilio
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00370]

Gene therapy is a potential treatment for severe inherited disorders for which there is little hope of finding a conventional cure. These include lethal diseases like immunodeficiencies and metabolic disorders, and non lethal conditions associated to poor quality of life and life-long symptomatic treatments, like muscular dystrophy, cystic fibrosis or thalassemia. Skin adhesion defects belong to both groups. For the non-lethal forms, gene therapy, or transplantation of cultured skin derived from genetically corrected epidermal stem cells, represents a very attractive therapeutic option, and potentially a definitive treatment. Recent advances in gene transfer and stem cell culture technology are making this option closer than ever. This paper critically reviews the progress and prospects of gene therapy for skin adhesion defects, and the factors currently limiting its development.
[Back to top]


Apoptotic pathways in the pathogenesis of pemphigus: targets for new therapies
Roberta Lotti, Alessandra Marconi and Carlo Pincelli
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/00371]

Pemphigus is a group of rare autoimmune blistering diseases of the skin in which autoantibodies to desmosome cadherins, desmogleins, induce loss of cell-cell adhesion (acantholysis). In addition to steric hindrance and activation of intracellular phosphorylation cascade signaling pathways, apoptosis has been suggested to contribute to the mechanism by which pathogenic IgG induces acantholysis. We review the literature examining the role of apoptosis in pemphigus. Current data recognize a central role of apoptosis in the mechanisms of blister induction. In particular, here we stress the key role of FasL in pemphigus, as it is able to first induce apoptosis, then acantholysis. Being pro-apoptotic molecules important in blister formation, they could represent new specific targets for pemphigus treatment.
[Back to top]


The role of an experimental model of atherosclerosis: apoE-knockout mice in developing new drugs against atherogenesis
Jacek Jawien
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003703]

Although atherosclerosis was previously thought to be mainly a degenerative disease, it is now well ascertained that its pathogenesis is inflammatory. There was a pivotal role of apoE-knockout mice in understanding the inflammatory background of atherosclerosis. Currently, atherosclerosis is known as a chronic inflammatory disease, in most cases initiated by hypercholesterolemia.

Since 1992 the mouse has become an excellent model for experimental atherosclerosis research. Until 1992, the diet-induced atherosclerosis mouse model has been used effectively, but the lesions tended to be small and were limited to early fatty-streak stage. This model was also criticized because of the toxicity and inflammatory responses due to the diet. In 1992 the first line of gene targeted animal models, namely apolipoprotein E-knockout mice was developed. Of the genetically engineered models, the apoE-deficient model is the only one that develops extensive atherosclerotic lesions on a chow diet. It is also the model in which the lesions have been characterized most thoroughly. The lesions develop into fibrous plaques; however, there is no evidence that plaque rupture occurs in this model. The LDL receptor - deficient model has elevated LDL levels, but no lesions, or only very small lesions, form on the chow diet, however, robust lesions do form on the western-type diet. The creation of apoE-knockout mice has changed the face of atherosclerosis research.

The apoE-deficient mouse model of atherosclerosis can then be used to: 1) identify atherosclerosis susceptibility modifying genes, by the candidate-gene and gene-mapping methods; 2) identify the role of various cell types in atherogenesis; 3) identify environmental factors affecting atherogenesis; and 4) assess therapies that might block atherogenesis or lesion progression.

ApoE-deficient mice have also been used to look for environmental and drug effects on atherosclerosis and to test novel therapies.

Gene-targeted mouse models has changed the face of atherosclerotic research and helped in creation of the new theory of atherosclerosis - as an inflammatory disease. Nowadays, apoE-knockout mice model is therefore used in developing new drugs against atherosclerosis. This review describes how new groups of agents are searched.
[Back to top]


Regeneration of Vessel Wall Functionality and Vascular Restoration Therapy with Biodegradable Stents – Current Status
Rafał Depukat, Łukasz Rzeszutko and Dariusz Dudek
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003704]

The novel vascular solution, the bioresorbable scaffold, has already been called Vascular Regeneration Therapy (VRT), providing a new quality in interventional cardiology. This new generation of stents gives a potential advantage over the permanent metal prosthesis, mainly restoration of vasomotion after full biodegradation. In the article we summarize the latest achievements in stent technologies allowing complete regeneration of arterial wall functions after implantation of biodegradable scaffold.
[Back to top]


How drugs influencing central blood pressure prevent atherosclerosis complications?
Piotr Jankowski, Michel E. Safar and Kalina Kawecka-Jaszcz
[Purchase Article] [BSP/CPB/E-Pub/003705]

Although the differences between central and peripheral blood pressure (BP) have been known for decades, the consequences of decision-making based on peripheral rather than central BP have only recently been recognized. Central PP is closer to the heart, coronary and carotid arteries, which are the most important sites of cardiovascular events. The influence of cyclic stretch (owing to cyclic changes in BP) on the arterial wall has been documented at every stage of atherosclerosis development. Apart from mediating atherosclerosis progression and plaque instability, the pulsatile component of BP is the main mechanism leading to plaque rupture and, consequently, to acute coronary syndromes and other vascular complications. Latest evidence suggests that the effect of some antihypertensive drugs on central BP (especially on central pulse pressure) is greater when compared with the effect on peripheral pressure, especially if systolic or pulse pressure are measured. Recently, a new group of drugs (advanced glycation end products cross-link breakers) were developed which have ability to decrease pulsatile component of blood pressure. The principal goal of the present review is to update the latest advances in this field.
[Back to top]


Novel therapeutic approaches in limiting oxidative stress and inflammation
Agnieszka Spychalowicz, Grzegorz Wilk, Tomasz Śliwa, Dominik Ludew and Tomasz J. Guzik
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003706]

The interaction between reactive oxygen species (ROS) and inflammation plays and important role in the pathogenesis of endothelial dysfunction and cardiovascular disease, cancer and other diseases. Thus, antioxidant strategies may be very important in immune regulation and in limiting inflammation. Surprisingly, large clinical trials have shown that ROS scavenging by antioxidant vitamins is ineffective or even harmful in spite of the fact that reactive oxygen species themselves are pro-inflammatory, regulate immune system and enhance atherosclerosis. Therefore, there is a need of novel, more specific antioxidant and anti-inflammatory approaches aimed on prevention of ROS formation, by targeting specific molecular pathways involved in ROS generation and their activation of pro-inflammatory cascades. Potential targets include the NADPH oxidases (Nox enzymes), xanthine oxidase, endothelial nitric oxide synthase and mitochondrial oxidases. Nox enzymes play central role, as they can stimulate other enzymatic sources of ROS. The interplay between inflammation and oxidative stress is discussed in the context of adipose tissue, perivascular inflammation and role of central nervous system in immune regulation. All of the above participate in “brain-vessel axis” critical in the pathogenesis of numerous pathologies. Role of cytokines such as TNF-alpha, IL-17 or IL-6 and their links to superoxide and hydrogen peroxide production are discussed.   Statins, angiotensin converting enzyme inhibitors and angiotensin II receptor antagonists block upstream signaling of Nox activation, including MAP kinase signaling or G protein activation, which contributes to their clinical effectiveness.  Here we discuss novel possibilities that drugs directly inhibiting Nox activation could successfully inhibit oxidative stress and inflammation related to cardiovascular disease. Moreover we describe potential gene therapy approaches in limiting oxidative stress in the vasculature. These approaches can become also useful in cancer immunomodulation.
[Back to top]


Genetics of hypertrophic and dilated cardiomyopathy
Felix W. Friedrich and Lucie Carrier
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003707]

Cardiomyopathies are categorized as extrinsic, being caused by external factors, such as hypertension, ischemia, inflammation, valvular dysfunction, or as intrinsic, which correspond to myocardial diseases without identifiable external causes. These so called primary cardiomyopathies can be categorized in four main forms: hypertrophic, dilated, restrictive, and arrhythmogenic right ventricular cardiomyopathy. Cardiomyopathies are diagnosed by clinical expression, echocardiography, electrocardiography, non-invasive imaging, and sometimes by cardiac catheterization to rule out external causes as ischemia. The two main forms of primary cardiomyopathies are the hypertrophic and dilated cardiomyopathies. Most of hypertrophic cardiomyopathy and 20-50% of dilated cardiomyopathy are familial showing a wide genetic and phenotypic heterogeneity. This review presents the current knowledge on the causative genes, molecular mechanisms and the genotype–phenotype relations of hypertrophic and dilated cardiomyopathies.
[Back to top]


Fabry Disease Cardiomyopathy: from genes to clinical manifestations
Attila Béla Kertész and István Édes
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003708]

Fabry disease is an X chromosome linked disorder caused by the inherited deficiency of lysosomal enzyme α-galactosidase A. The deficiency results in abnormal degradation of certain glycosphingolipids. Although the disease is known for more than hundred years and the underlying molecular basis is getting to be well defined, there are still a lot of unanswered questions regarding the different clinical presentations, available diagnostic procedures and therapeutic interventions. The scope of the article is to review the molecular basis of Fabry disease and summarize the available data about Fabry disease cardiomyopathy, highlight the controversies of current knowledge and evaluate future research directions.
[Back to top]


Understanding the molecular and cellular changes behind aortic valve stenosis
Inês Falcão-Pires, Cristina Gavina and Adelino F. Leite-Moreira
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003709]

Morbidity from degenerative aortic valve disease (AVS) is increasing worldwide, concomitant with the ageing of the population and the growing consumption of high caloric and cholesterol diets of the western countries.

Despite the high prevalence of AVS, with its high mortality and morbidity, studies on the molecular and cellular mechanisms underlying the onset of aortic valve degeneration have only advanced in the last 15 years. The result of this effort is now beginning to reveal several mechanisms with great therapeutic targeting potential that may alter the natural history of this progressive pathology. Indeed, the view of this disease has changed from being an unmodifiable degenerative disease to an active biological process regulated by highly conserved cellular pathways.

The progression of AVS includes inflammation, angiogenesis and remodelling of the extracellular matrix leading to osteogenesis in the aortic valve and revealing many mechanisms and risk factors similar to atherosclerosis. Therefore statins and angiotensin II antagonists seemed promising treatment options; however, experimental results are still controversial. Nonetheless, valvular degeneration results in dramatic myocardial changes induced by chronic pressure overload such as left ventricular hypertrophy as well as other paramount myocardial extracellular changes.

Currently, a strong impulse for future research to investigate the pathophysiological mechanisms and their modulation in order to prevent/delay the onset or progression of valve degeneration is needed. In the present review, we focused on the molecular and cellular mechanisms underlying degenerative AVS and its myocardial impact.
[Back to top]


Pharmacotherapy of aortic stenosis – success or failure ?
Pawel Petkow Dimitrow, Marek Jawien and Renata Rajtar-Salwa
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003710]

Aortic stenosis (AS) is a degenerative valvular disease that has, until recently, been considered to be a progressive and the most importantly unmodifiable process. However, recent histopathologic studies have clearly reported that the both development and progression of AS is based on an biochemically active process which is mediated by a chronic inflammation (featured by many similarities with atherosclerosis) and includes inflammatory-cell infiltrates, lipoproteins, lipids and factors responsible for calcification (extracellular-bone-matrix proteins and bone mineral). Targeted drug therapy to prevent the progression of calcific AS disease should ideally be based on the knowledge of risk factors and the molecular pathogenesis of the disease. Treatment trials using various drugs have been undertaken to test whether medical therapy (statins, renin-angiotensin inhibition and anti-osteoporosis drugs) can prevent or beneficially modify the disease course. Although retrospective and non-randomized studies suggested a positive effect of statins, benefit has not been seen in perspective randomized controlled trials. Inhibition of renin-angiotensin has shown discordant results in retrospective studies with no randomized controlled data published. In the future, we need to consider other medical therapies (anti-osteoporosis drugs are attractive candidates) that might target different pathways in this disease process. Additionally, we need to detect the optimal moment to start statin therapy for this chronic slowly progressive disease; treatments aimed at the early disease process (pre-obstructive form of aortic sclerosis) may be ineffective with end-stage tissue changes.
[Back to top]


Left ventricular hypertrophy in isolated aortic stenosis: primetime for the ventricle
Cristina Gavina, Inês Falcão-Pires, Francisco Rocha-Gonçalves and Adelino Leite-Moreira
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003711]

Aortic stenosis is the most common type of valvular heart disease and its recent increase is related to aging. The decreased aortic valve area imposes a chronic systolic pressure overload to the left ventricle. In response, the ventricle hypertrophies in an attempt to normalize the increased wall stress, but this response is not uniform in patients with similar degrees of stenosis and its regression after surgical correction is variable, suggesting that several factors, other than load, can explain these differences. These findings are particularly important since the presence of left ventricular hypertrophy after aortic valve replacement is an independent predictor of worse outcome, probably because it indicates irreversible remodeling. Age, gender, hypertension, patient-prosthesis mismatch and interstitial remodeling also play an important role in this setting, raising the possibility of intervention beyond valve replacement. The possibility of combining estrogen treatment, antihypertensive agents, antioxidants and modulators of the renin-angiotensin-aldosterone system with surgical treatment to promote reverse remodeling is very appealing. On the other hand, a preventive strategy to intervene earlier in patients with significant left ventricular mass and avoid patient-prosthesis mismatch, especially in the younger and those with systolic dysfunction, can have a significant impact on prognosis. Further evidence, with well designed clinical trials, is needed but the spotlight must be in the ventricle, not the valve.
[Back to top]


How cardiomyocytes make the heart old
Zoltán Papp, Dániel Czuriga, László Balogh, Ágnes Balogh and Attila Borbély
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003712]

Naturally occurring decline in cardiovascular reserve with age associates with a combination of the reduction in cardiomyocyte number and altered cardiomyocyte function. Recent investigations suggested that about half of the cardiomyocytes is the same as at birth, while the other half of the cardiomyocytes is the result of cardiomyocyte renewal in the senescent heart. In addition, the total number of cardiomyocytes is estimated to be less by one third in the old heart than the number of cardiomyocytes at birth. Thus, the reduction in cardiomyocyte number of the aging heart cannot be fully compensated by cardiomyocyte renewal. Aging of long-lived differentiated myocardial cells, as well as of cardiac progenitor stem cells may contribute to an increased rate of apoptosis, and decreased capacity of cell duplication and/or differentiation. In addition, differentiated cardiomyocytes are prone for accumulating biological by-products of cellular metabolism and of incompletely processed oxidative insults. In this context, interactions between lysosomes and mitochondria may provide a mechanistic background for the age-dependent alterations in cardiac macromolecules. This reasoning postulates a direct relationship between the number of pro-oxidative, ill-functioning mitochondria and the amount of ballast-overloaded lysosomes in long-lived cardiomyocytes. Accumulation of biological garbage and telomere shortening might be considered as hallmarks of cardiomyocyte aging with implications for depressed cardiac function and cardiomyocyte renewal. Changes in protein expression together with posttranslational modifications of myocardial proteins affect excitation-contraction coupling and explain the declining mechanical function of the cardiomyocytes. Altogether, these changes represent a significant part of the reduced cardiovascular reserve in aged individuals.
[Back to top]


Alteration of the beta-adrenergic signaling pathway in human heart failure
Nazha Hamdani and Wolfgang A. Linke
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003713]

Heart failure is characterized by elevated circulating catecholamine levels and extensive abnormalities in β-adrenergic receptor signaling. Under physiological conditions, sympathetic modulation via catecholamines induces positive inotropic, chronotropic and lusitropic responses. Well established in heart failure patients is a pronounced activation of the sympathetic system, accompanied by downregulation and desensitization of β-adrenergic receptors, leading to a markedly diminished β-adrenergic contractile response. In this review, we will discuss the normal β-adrenergic receptor signaling pathway in the heart and focus on the pathphysiological alterations of β-adrenergic receptor signaling and contractile proteins in heart failure.
[Back to top]


Cellular mechanisms for diastolic dysfunction in the human heart
Dániel Czuriga, Walter J. Paulus, István Czuriga, István Édes, Zoltán Papp and Attila Borbély
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003714]

Left ventricular (LV) diastolic dysfunction is an important contributor to many different cardiovascular diseases. LV diastolic dysfunction can manifest itself as slow LV relaxation, slow LV filling or high diastolic LV stiffness. Diastolic abnormalities have been described in the senescent heart, in heart failure with normal ejection fraction (HFNEF), in diabetic cardiomyopathy, in aortic valve stenosis (AVS), in hypertrophic cardiomyopathy (HCM), as well as in Fabry disease (FD), however, exact cellular and molecular alterations behind the diastolic deterioration in these diseases are not yet completely characterized. Several studies thoroughly investigated altered cardiomyocyte function, changes of contractile myofilaments, extracellular collagen deposition and advanced glycation end products (AGEs) cross-linking in the background of diastolic dysfunction. These clinical and experimental data suggest that underlying mechanisms of LV diastolic dysfunction are divergent in different cardiac pathologies, therefore the present review aims to summarize mechanisms at the cellular level of diastolic abnormalities in various cardiovascular diseases.
[Back to top]


Diastolic heart failure and LV dyssynchrony
Mario Kasner, Dirk Westermann, Heinz-Peter Schultheiss and Carsten Tschöpe
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003714]

Our knowledge of diastolic heart failure (DHF) is still limited with regard to pathophysiology, diagnosis and clinical treatment. Amongst others, LV dyssynchrony was suggested to be an additional factor involved in the pathogenesis of subgroup of patients with DHF. In 20-30% of patients with DHF a systolic LV dyssynchrony could be detected and about 20% DHF patients evidenced a diastolic dyssyncrony. Both systolic and diastolic dyssynchrony may contribute to the impairment of cardiac function and clinical manifestation in DHF. Opposite to the systolic heart failure, wide QRS complex is uncommon which incriminates that dyssynchrony in DHF is rather related to regional disperse in contractility than to electromechanical coupling delay. Asynchronous LV relaxation and impairment of ventricular restoring forces may also impair the LV filing and lead to a diastolic dyssynchrony. Particularly in patients with preserved LV contractility mechanical LV dyssynchrony induces energy wastage and consequently reduces cardiac reserves. However, up to date it is not clear to what degree LV dyssynchrony is involved in the pathomechanisms of this subpopulation of DHF.
[Back to top]


Development of Stable Lyophilized Protein Drug Products
Richard L. Remmele, Sampathkumar Krishnan and William J. Callahan
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003716]

Freeze drying, or lyophilization is widely used for biopharmaceuticals to improve the long term storage stability of labile molecules. This review examines general theory and practice of rational lyophilization of biopharmaceuticals. Formulation development involving the selection of appropriate excipients, their associated physical properties, and mechanism of action in achieving a stable drug product are primary considerations for a successful lyophilization program. There are several parameters considered critical on the basis of their relationship to lyophilization cycle development and protein product stability. This along with the importance of analytical methods to provide insight toward understanding properties of drug product stability and cake structure are discussed. Also, aspects of instability found in lyophilized biopharmaceutical products, their degradation pathways and control are elucidated. Finally, container-closure requirements and drug product handling are described in context of the caveats to avoid compromising drug product quality.
[Back to top]


Bacteria and their toxins tamed for immunotherapy
Irena Adkins, Jana Holubova, Martina Kosova and Lenka Sadilkova
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003718]

Bacterial toxins share the ability to enter host cells to target various intracellular proteins and to modulate host immune responses. Over the last 20 years, toxins and their mutated variants, as well as live attenuated bacteria, have been exploited for vaccination and immunotherapy of various infectious, malignant and autoimmune diseases. The ability of Bordetella pertussis adenylate cyclase toxin to translocate its adenylate cyclase domain across the host cell membrane, as well as the pathways of intracellular trafficking of Bacillus anthracis lethal and edema toxins, Shigella dysenteriae shiga toxin or Escherichia coli shiga-like toxin,have been repeatedly exploited for the delivery of antigenic epitopes into host cells and for stimulation of antigen-specific T cell responses. Similarly, E. coli α-hemolysin, or effector proteins of Yersinia and Salmonella secreted by the type III secretion systems, were used to facilitate the delivery of fused heterologous proteins or peptides for antigenic presentation. Vibrio cholerae cholera toxin, E. coli heat-labile enterotoxin, B. pertussis pertussis toxin or the Cry1A protein of Bacillus thuringiensis have shown a great potential to act as adjuvants and to stimulate mucosal as well as systemic immune responses. The immunotherapeutic potential of some toxins, like Clostridium perfringens perfringolysin O, Streptococcus intermedius intermedilysin, or Streptococcus pneumoniae pneumolysin needs to be evaluated further. The Bordetella adenylate cyclase toxoid used as a vaccine delivery tool, or Corynebacterium diphtheriae diphtheria toxin and Pseudomonas aeruginosa exotoxin A-based immunotoxins, are currently in various phases of clinical trials for cancer immunotherapy, as are some antigen-delivering Salmonella and Listeria monocytogenes strains.
[Back to top]


Clinical Experience with Gene Therapy and Bispecific Antibodies for T Cell-based Therapy of Cancer 
Patrick A. Baeuerle and Christian Itin
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003719]

Unlike any other cell type, T cells have a unique potential to eliminate cancer cells and to eventually cure cancer patients. As a result, researchers have made extensive efforts over the past three decades to develop therapeutics with the potential to mount T cell responses against cancer cells. One way in which such T cell responses can be triggered is by vaccines and adjuvants, potentially leading to tumor-specific T cell clones and lasting immunity. Alternatively, they can be induced with the help of recombinant proteins that either are expressed in patients’ T cells by gene therapeutic means, or are delivered to patients as pharmaceuticals for temporary engagement of T cells. With both recombinant technologies, cytotoxic T cells can be engaged for cancer cell lysis regardless of T cell receptor specificity and with the prospect of bypassing both complex T cell regulation and frequent immune escape mechanisms of tumor cells. In this review, we will focus on recombinant approaches for T cell engagement that currently are in clinical development. Approaches transfecting patient T cells with genes encoding recombinant T cell receptors or antibody fusion proteins will be compared to those temporarily engaging T cells by infused recombinant bispecific proteins. Initial experience has recently been gained in the clinic with both technologies such that their fundamental differences can now be discussed on the basis of patient data.
[Back to top]


Current Immunotherapy of Multiple Sclerosis and Future Challenges: Relevance of Immune-mediated Repair
Michael D. Carrithers
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003720]

Multiple sclerosis is the most common chronic inflammatory disease of the central nervous system and can cause severe neurological disability. Current immune therapy with beta-interferons, glatiramer acetate, immune suppressives, or selective adhesion molecule inhibitors can reduce the frequency and severity of acute attacks in a majority of patients but have little effect on the progressive phase of the disease.  Patients who require aggressive immune therapy are also at risk for the development of potentially fatal opportunistic infections.  Here current and emerging immune therapy options are discussed, and immune-mediated strategies to preserve normal immune surveillance and mediate tissue repair are proposed.
[Back to top]


Infections in patients with rheumatic diseases treated with TNF antagonists
Beatriz Pérez-Zafrilla, Loreto Carmona and Juan J Gómez-Reino
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003721]

TNF has a critical role in inflammation and immunity, and therapeutic inhibition of TNF with antagonist could potentially lead to immune suppression. Data gathered from clinical trials and clinical observation show a minor but significant increased risk of infections in patients suffering from rheumatic diseases treated with monoclonal TNF antibodies and soluble TNF receptors. This increase risk applies to patients but also to the underlying disease. Pathogens causing these infections include intracellular bacteria, and to some extend opportunistic microorganisms as mycobacteria and fungi. Preventive strategies and patient selection have an important impact on the risk of these infections.
[Back to top]


Monoclonal Antibodies For The Immunotherapy Of Solid Tumours
Richard Mauerer and Rudolf Gruber
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003722]

More than 80% of all cancers are caused by solid malignancies. More than 90% of these tumours are of epithelial origin. The main principles in tumour treatment are surgery, radiotherapy, chemotherapy or combinations of these. Complete surgical removal of the tumour is the most effective therapy for solid malignancies. Recent advances in early cancer detection led to a higher rate of resectable primary tumours and therefore prognosis will be determined especially by metastasis. Even in early stages some tumour cells may disseminate for example into the bone marrow. If these occult metastasis get evident they are mostly incurable by surgery and often highly resistant to chemotherapy. Developing new therapeutic agents to destroy these resting cells is a major challenge for the improvement of cancer therapy in the future. Advances to reach these goals were made in immunological therapies with monoclonal antibodies (MABs). These MABs are for example directed against tumor-associated antigens (TAAs). By binding selectively on tumor cells they can activate immunological effector mechanisms, e.g. antibody dependent cell cytotoxicity (ADCC) or complement mediated lysis. Other mechanisms are the blocking of inhibiting molecules to re-activate anergic tumor infiltrating lymphocytes (TILs). Furthermore growing tumours depend on oxygen supply, i.e. on effective neovascularisation. Antibodies against VEGF are able to inhibit neovascularisation and are therefore used successfully in tumour therapy.
[Back to top]


The intrinsic apoptosis pathways as a target in anticancer therapy
Verena Jendrossek
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003723]

Tumor cells need to disrupt apoptosis pathways to escape the cytotoxic action of oncogene activation and microenvironmental stress during the carcinogenic process. However, the cytotoxic action of classical chemotherapy, and radiotherapy includes the induction of apoptotic cell death. Therefore, apoptosis resistance of tumor cells contributes to the failure of chemotherapy and radiotherapy.

During the last two decades, extensive research aimed at an improved understanding of the complex signaling pathways that control apoptosis execution in normal cells and of the endogenous factors that mediate apoptosis resistance of cancer cells. Among these, the Bcl-2 protein family attracted major attention for the development of compounds that specifically target apoptosis resistance of cancer cells. Bcl-2 proteins are master regulators of the intrinsic apoptosis pathway that is crucial for apoptosis execution in response to DNA-damage. The review will highlight current knowledge on the regulation of apoptosis pathways and discuss several approaches that target the Bcl-2 rheostat to counteract tumor cell intrinsic apoptosis resistance that may therefore be of value for a biological modulation of apoptosis resistance.
[Back to top]


Recent Advances in Active Specific Cancer Vaccine Treatment for Colorectal Cancer
Kiyotaka Okuno, Fumiaki Sugiura, Kyogo Itoh, Koji Yoshida, Takuya Tsunoda and Yusuke Nakamura
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003724]

Cloning techniques to identify genes and peptides of tumor-associated antigens have created new possibilities for the immunotherapy of patients with advanced cancer. Here, we review recent clinical trials of specific cancer vaccines, mainly HLA-restricted peptides, and epitope-encoding vectors for advanced colorectal cancer (CRC). Many researchers initially focused on carcinoembryonic antigen (CEA) as an immunologic target antigen that is overexpressed on virtually all CRCs. A recombinant vaccine containing the CEA gene and dendritic cells (DCs) loaded with CEA peptide was administered to patients with CEA-elevated CRC. Although CEA-specific responses were detected, the clinical responses were limited. Recently, new types of clinical trials—namely, a personalized protocol to take into account the immunological diversity of cytotoxic T cell responses among patients and a novel cancer-testis antigen protocol that uses multiple peptides derived from genes identified by the cDNA array method—have been introduced. The personalized protocol seemed to be better than the classical (non-personalized) protocol in terms of clinical response and survival. Novel cancer-testis antigen protocols that use multiple CRC-derived peptides were recently conducted in patients with advanced CRC. The preliminary study yielded promising results regarding specific T cell responses to peptides and survival benefits. In this review, we summarize these results and discuss future perspectives.
[Back to top]


Dendritic cells: elegant arbiters in human reproduction
Sabine e. Segerer, Claudia Rennemeier, Ulrike Kaemmerer, Torsten Frambach, Johannes Dietl and Lorenz Rieger
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003725]

The female reproductive tract represents a great challenge to the residing immune cells: Concomitantly, those immune-competent cells have to provide tolerogenic mechanisms favoring the development of a successful pregnancy while permitting protection against harmful pathogens. The predominant cell population facing this “double edged” regulatory capacity within the reproductive tract is that of dendritic cells (DC). There is evidence that DC represent a highly adaptive cell type, which can either be transformed in an immune-stimulatory phenotype after exposure to inflammatory or infectious signals, or in an immune inhibitory phenotype preventing T cell activation when located in an adequate anti-inflammatory microenvironment. Thus, this review highlights this two-faced character of DC focusing on their morphology and function within the human reproductive tract and especially during pregnancy.
[Back to top]


Dye Visualization – A Method for Investigating Biomechanical Flows
Thomas Leweke
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003727]

Flow visualization using dye is an inexpensive and easy-to-implement experimental technique. It can be used for a rapid qualitative assessment of fluid flows in configurations relevant to biomedical or biotechnological applications, which often involve small spatial dimensions and flow velocities (low Reynolds numbers). This paper gives an overview of the practical aspects related to dye visualization in liquids (dyes, introduction of dye into the flow, illumination), and discusses the information that can be obtained by this method, which includes the distribution of coherent structures/vortices, the location of recirculation zones, and certain characteristic spatial and temporal scales. Visualization results for three examples of generic flows related to biomechanical applications are presented: the flow behind a contraction in a pipe (stenosis), the wake of a particle moving along a wall, and the flow inside a lid-driven mixing vessel (bioreactor).
[Back to top]


Functional Imaging to Understand Biomechanics: A Critical Tool For the Study of Biology, Pathology and the Development of Pharmacological Solutions
R. Aidan Jamison, James A. Armitage, Josie Carberry, Marcus J. Kitchen, Stuart B. Hooper and Andreas Fouras
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003728]

We present four case studies of the literature discussing the effects of physical forces on biological function. While the field of biomechanics has existed for many decades, it may be considered by some a poor cousin to biochemistry and other traditional fields of medical research. In these case studies, including cardiovascular and respiratory systems, we demonstrate that, in fact, many systems historically believed to be controlled by biochemistry are dominated by biomechanics. We discuss both the previous paradigms that have advanced research in these fields and the changing paradigms that will define the progressions of these fields for decades to come. In the case of biomechanical effects of flowing blood on the endothelium, this has been well understood for decades. In the cases of platelet activation and liquid clearance from the lungs during birth, these discoveries are far more recent and perhaps not as universally accepted. While only a few specific examples are examined here, it is clear that not enough attention is paid to the possible mechanical links to biological function. The continued development of these research areas, with the inclusion of physical effects, will hopefully provide new insight into disease development, progression, diagnosis and effective therapies.
[Back to top]


Patient-Specific Modeling and Multi-Scale Blood Simulation for Computational Hemodynamic Study on the Human Cerebrovascular System
Marie Oshima,  Ryo Torii,  Shigefumi Tokuda, Shigeki Yamada and Akio Koizumi
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003729]

To develop a targeted drug delivery system for cerebrovascular disorders such as stroke, it is important to obtain detailed information on flow rates and hemodynamics of the human cerebrovascular system for individual patients. A patient-specific integrated numerical simulation system has been developed by the authors such that vascular geometry is constructed from medical images such as magnetic resonance imaging (MRI) or computed tomography (CT) data, and computational conditions are modeled mathematically to represent the realistic in vivo environments.

In general, the three-dimensional numerical simulation using a patient-specific model is conducted only for a localized diseased region with atherosclerosis or an aneurysm. Although the analysis region is only a part of the circulatory system, the simulation should include the effects from the entire circulatory system.  Since the peripheral network determines the flow distributions in the cerebrovascular system, the paper reviews the recent simulation methods to take into account the network by coupling the image-based three-dimensional simulation with a one- and zero-dimensional simulations as an outflow boundary condition The paper shows the mathematical modeling of the multi-scale outflow boundary condition and its applications to patient-specific models of the arterial circle of Willis. The results are compared to those using the conventional, free-stream boundary condition. As a result, the multi-scale outflow boundary condition shows a significant difference in flow rate of each artery and in flow distribution in the arterial circle of Willis.
[Back to top]


Recent software developments and applications in functional imaging
Lingfeng Wen, Stefan Eberl, Michael Fulham and (David) Dagan Feng
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003730]

Functional imaging allows the quantification of biochemical or biophysiological changes in-vivo through the visualization of the spatial distribution and temporal changes of administrated radiopharmaceuticals. Instrumentation advances such as PET-CT (positron emission tomography - computed tomography) and PET-MR (positron emission tomography - magnetic resonance), improvements in image processing and reconstruction, the development of target and disease-specific radiotracers and improved kinetic modelling techniques, have substantially enhanced our ability to measure functional changes in normal and diseased states. Various combinations of these advances and refinements are now used in routine clinical practice for patient care. In this paper we review recent literature on software developments and applications in image restoration, motion correction, kinetic analysis, and image processing in the field of functional imaging.
[Back to top]


Ultrasound: Medical Imaging and Beyond
Haim Azhari
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003731]

Medical applications of ultrasound were first investigated about seventy years ago.  It has rapidly evolved since then, becoming an essential tool in medical imaging. Ultrasound ability to provide real time images with frame rates exceeding several hundred frames per second allows one to view rapid anatomical changes as well as to guide minimal invasive procedures. By, combining Doppler techniques with anatomical images ultrasound provides real time quantitative flow information as well. It is portable, versatile, cost effective and considered sufficiently hazardless to monitor pregnancy. Moreover, ultrasound has the unique capacity to offer therapeutic capabilities in addition to its outstanding imaging abilities.  It can be used for physiotherapy, lithotripsy, and thermal ablation, and recent studies have demonstrated its usefulness in drug delivery, gene therapy and molecular imaging.
[Back to top]


Opto-acoustic Imaging of Drug Discovery Biomarkers
Bohumil Bednar and Vasilis Ntziachristos
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003732]

Optical imaging has seen significant developments over the past decade as an investigational tool for in-vivo visualization of cellular and sub-cellular events. With the recent addition of optoacoustic (photoacoustic) methods, in particular multi-spectral opto-acoustictomography (MSOT), to the already rich armamentarium of photonic methods the capacity of optical molecular imaging across scales haswidened significantly. MSOT brings unique features into optical imaging, namely high resolution optical imaging over several millimeters to centimeters of tissue depth and the ability to simultaneously resolve multiple tissue molecules and extrinsically administered optical or optoacoustic agents with physiological or molecular specificity. Here, we discuss the implications of utilizing MSOT in the context of drug discovery and review suitable optoacousticagents against disease and drug efficacy biomarkers. The combination of existing knowledge on generating optical targeted contrast, with the high resolution deep tissue visualization offered by MSOT, allows for the development of next-generation biological optical imaging and corresponding drug discovery applications.
[Back to top]


Generation of Human single-chain Antibody to the CD99 Cell Surface Determinant Specifically Recognizing Ewing’s Sarcoma Tumor Cells
Mara Gellini, Alessandro Ascione, Michela Flego, Alessandra Mallano, Maria Luisa Dupuis, Silvia Zamboni, Manuela Terrinoni, Valeria D’Alessio, Maria Cristina Manara, Katia Scotlandi, Piero Picci and Maurizio Cianfriglia
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003733]

The survival of pediatric patients with cancer entities including osteosarcoma and Ewing's sarcoma (ES), remains extremely low hence novel treatment approaches are urgently needed. Therefore, based on the concept of targeted therapy, numerous potential targets for the treatment of these cancers have been evaluated pre-clinically or in some cases even clinically during the last decade. In ES the CD99 protein is an attractive target antigen. In this respect, a new entry site for therapeutic intervention may derive from specific human antibodies against CD99.

Human scFvC7 was isolated from a semi-synthetic ETH-2 antibody phage library panned on the extracellular portion of recombinant human CD99 protein. The scFvC7 was genetically sequenced, tested for CD99 recognition on an array of recombinant CD99 fragments and measured for binding affinity by ELISA. Finally, it was tested for staining CD99 antigen on a large panel of tumor and normal cells and tissues by cytofluorimetric and immunohystochemical assays.

The new antibody scFvC7 recognizes the CD99 extracellular domain included between residues 50 and 74 with a binding affinity of 2.4 x 10^-8 M. In contrast with all other antibodies to CD99 so far isolated, scFvC7 shows a unique specificity in cancer cell recognition: It stained prevalently ES cells while no or weak reactivity was observed on the majority of the other tumor and normal cells and tissues.
[Back to top]


Fluorescence Detection of MMP-9.II. Ratiometric FRET-Based Sensing With Dually Labeled Specific Peptide
Rafal Fudala,Ryan Rich,Anindita Mukerjee, Amalendu P. Ranjan,Jamboor K. Vishwanatha, Anna K. Kurdowska, Zygmunt Gryczynski, Julian Borejdo and Ignacy Gryczynski
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003734]

In our previous paper we showed that theMMP-9 enzyme recognizes a specific peptide sequence,Lys-Gly-Pro-Arg-Ser-Leu-Ser-Gly-Lys,and cleaves the peptide into two parts [1]. In this study, the peptide is labeledwith two dyes, carboxyfluorescein (5-FAM) and Cy5. A highly efficient energy transfer of over 80% results in a dominant emission of Cy5 at ~670 nm with an excitation of 470 nm. Severance of the peptide by the MMP-9 enzyme eliminatesFörster Resonance Energy Transfer (FRET) and strongly increases the fluorescence of the 5-FAM dye.In this manuscript we describe the strategy for a FRET-based method for MMP-9 enzyme detection. The basic aim is to apply a ratio-metric sensing technique in which a ratio of green/red fluorescence intensity is measured as a function of enzyme concentration. The ratio-metric method eliminates many experimental variables and enables accurate MMP-9 detection.
[Back to top]


Antidote-Control Of Aptamer Therapeutics: The Road To A Safer Class Of Drug Agents
K.M. Bompiani, R.S. Woodruff, R.C. Becker, S.M. Nimjee and B.A. Sullenger
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003736]

Aptamers, or nucleic acid ligands, have gained clinical interest over the past 20 years due to their unique characteristics, which are a combination of the best facets of small molecules and antibodies. The high binding affinity and specificity of aptamers allows for isolation of an artificial ligand for theoretically any therapeutic target of interest. Chemical manipulations of aptamers also allow for fine-tuning of their bioavailability, and antidote control greatly expands their clinical use. Here we review the various methods of antidote control of aptamer therapeutics – matched oligonucleotide antidotes and universal antidotes. We also describe the development, recent progress, and potential future therapeutic applications of these types of aptamer-antidote pairs.
[Back to top]


Black Widow Spider (Latrodectus Mactans) Antivenom In Clinical Practice
Andrew A. Monte
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003737]

Black widow spider (Latrodectus mactans) envenomation has been recognized since antiquity. The syndrome, latrodectism, is characterized by painful muscle rigidity and autonomic disturbances such as tachycardia, hypertension, and diaphoresis. Symptoms typically last for 1-3 days. Treatment has ranged from local folk remedies to administration of specific antivenom. Opioid analgesics combined with muscle relaxants, such as benzodiazepines, are only effective at symptomatic and temporary control. Antivenom is by far the most efficacious therapy available based on symptom resolution, need for subsequent therapy, and hospital admission rates. Fear of allergic type reactions from antivenom administration has limited its use in the United States. A new purified F(ab)2 fragment Latrodectus mactans antivenom, Analatro®, is currently undergoing clinical trials. The product is expected to have similar efficacy and be associated with fewer adverse reactions when compared to the currently available partially purified whole IgG Merck product. This shift in the risk-benefit analysis may ultimately lead to more antivenom administration in significantly envenomated patients.
[Back to top]


Antidotes For Acute Cyanide Poisoning
Stephen W. Borron and Frederic J. Baud
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003738]

Cyanide poisoning can present in multiple ways, given its widespread industrial use, presence in combustion products, multiple physical forms, and chemical structures. The primary target of toxicity is mitochondrial cytochrome oxidase. The onset and severity of poisoning depend on the route, dose, physicochemical structure and other variables. Common poisoning features include dyspnea, altered respiratory patterns, abnormal vital signs, altered mental status, seizures, and lactic acidosis. Our present knowledge supports cyanide poisoning treatment based on excellent supportive care with adjunctive antidotal therapy. Multiple antidotes exist and vary in regional availability. All currently marketed antidotes appear to be effective. Antidotal mechanisms include chelation, formation of stable, less toxic complexes, methemoglobin induction, and sulfane sulfur supplementation for detoxification by endogenous rhodanese. Each antidote has advantages and disadvantages. For example, hydroxocobalamin is safer than the methemoglobin inducers in patients with smoke inhalation. Research for new, safer and more effective cyanide antidotes continues.
[Back to top]


Dexrazoxane For The Prevention Of Cardiac Toxicity And Treatment Of Extravasation Injury From The Anthracycline Antibiotics
James H. Doroshow
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003739]

The cumulative cardiac toxicity of the anthracycline antibiotics and their propensity to produce severe tissue injury following extravasation from a peripheral vein during intravenous administration remain significant problems in clinical oncologic practice. Understanding of the free radical metabolism of these drugs and their interactions with iron proteins led to the development of dexrazoxane, an analogue of EDTA with intrinsic antineoplastic activity as well as strong iron binding properties, as both a prospective cardioprotective therapy for patients receiving anthracyclines and as an effective treatment for anthracycline extravasations. In this review, the molecular mechanisms by which the anthracyclines generate reactive oxygen species and interact with intracellular iron are examined to understand the cardioprotective mechanism of action of dexrazoxane and its ability to protect the subcutaneous tissues from anthracycline-induced tissue necrosis.
[Back to top]


Calcium And Zinc Dtpa Administration For Internal Contamination With Plutonium-238 And Americium-241
Ziad N. Kazzi, Alexander Heyl and Dr. Johann Ruprecht
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003740]

The accidental or intentional release of plutonium or americium can cause acute and long term adverse health effects if they enter the human body by ingestion, inhalation, or injection. These effects can be prevented by rapid removal of these radionuclides by chelators such as calcium or zinc diethylenetriaminepentaacetate (calcium or zinc DTPA). These compounds have been shown to be efficacious in enhancing the elimination of members of the actinide family particularly plutonium and americium when administered intravenously or by nebulizer. The efficacy and adverse effects profile depend on several factors that include the route of internalization of the actinide, the route, type, and time of administration of the chelator, and whether the calcium or zinc salt of DTPA is used. Current and future research efforts should be directed at overcoming limitations associated with the use of these complex drugs by using innovative methods that can enhance their structural and therapeutic properties.
[Back to top]


Legalon® Sil: The Antidote Of Choice In Patients With Acute Hepatotoxicity From Amatoxin Poisoning
Ulrich Mengs, Ralf -Torsten Pohla and Todd Mitchellb
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003741]

More than 90% of all fatal mushroom poisonings worldwide are due to amatoxin containing species that grow abundantly in Europe, South Asia, and the Indian subcontinent. Many cases have also been reported in North America. Initial symptoms of abdominal cramps, vomiting, and a severe cholera-like diarrhea generally do not manifest until at least six to eight hours following ingestion and can be followed by renal and hepatic failure. Outcomes range from complete recovery to fulminant organ failure and death which can sometimes be averted by liver transplant. There are no controlled clinical studies available due to ethical reasons, but uncontrolled trials and case reports describe successful treatment with intravenous silibinin (Legalon® SIL). In nearly 1,500 documented cases, the overall mortality in patients treated with Legalon® SIL is less than 10% in comparison to more than 20% when using penicillin or a combination of silibinin and penicillin. Silibinin, a proven antioxidative and anti-inflammatory acting flavonolignan isolated from milk thistle extracts, has been shown to interact with specific hepatic transport proteins blocking cellular amatoxin re-uptake and thus interrupting enterohepatic circulation of the toxin. The addition of intravenous silibinin to aggressive intravenous fluid management serves to arrest and allow reversal of the manifestation of fulminant hepatic failure, even in severely poisoned patients. These findings together with the available clinical experience justify the use of silibinin as Legalon® SIL in Amanita poisoning cases.
[Back to top]


Reactive Skin Decontamination Lotion (Rsdl) For The Decontamination Of Chemical Warfare Agent (Cwa) Dermal Exposure
Schwartz, Hurst, Kirk, Reedy and Braue EH Jr
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003742]

Rapid decontamination of the skin is the single most important action to prevent dermal absorption of chemical contaminants in persons exposed to chemical warfare agents (CWA) and toxic industrial chemicals (TICs) as a result of accidental or intentional release. Chemicals on the skin may be removed by mechanical means through the use of dry sorbents or water. Recent interest in decontamination systems which both partition contaminants away from the skin and actively neutralize the chemical has led to the development of several reactive decontamination solutions. This article will review the recently FDA-approved Reactive Skin Decontamination Lotion (RSDL) and will summarize the toxicity and efficacy studies conducted to date. Evidence of RSDL’s superior performance against vesicant and organophosphorus chemical warfare agents compared to water, bleach, and dry sorbents, suggests that RSDL may have a role in mass human exposure chemical decontamination in both the military and civilian arenas.
[Back to top]


Antivenoms For Snakebite: Design, Function, And Controversies
Eric J. Lavonas
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003743]

Background: Animal-derived antivenoms have been used to treat snake envenomation for more than 100 years. Major technological advantages in the past 30 years have produced antivenoms that are highly purified and chemically modified to reduce the risk of acute hypersensitivity reactions. Like all pharmaceutical manufacture, commercial-scale antivenom production requires making trade-offs between cost, purity, pharmacokinetic profile, and production yield.

Scope: This article reviews the current state of the art for antivenom production and development. Particular attention is paid to controversies and trade-offs used to achieve a balance between improved safety and pharmacokinetic performance.
[Back to top]


Acetylcysteine Therapy for Acetaminophen Poisoning
Kennon Heard and Jody Green
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003744]

Acetylcysteine has been used to treat acetaminophen overdose for nearly 50 years. While no placebo controlled trials have been conducted, the efficacy of acetylcysteine is accepted for the prevention of hepatic injury when administered early after acetaminophen overdose. Acetylcysteine can be administered as an infusion or oral solution. The duration of treatment varies from 21 to 72 hours, depending on the protocol. Acetylcysteine also prevents death when administered to patients with hepatic failure from acetaminophen.
[Back to top]



Biomedical applications of molecular vibrational imaging “Metallic nanoparticles as SERS agents for biomolecular imaging”
Jun Ando and Katsumasa Fujita
[Abstract] [FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003746]

Raman spectroscopy is a promising technique for the identification and analysis of molecules in a sample without any labeling or modification. Since Raman scattering spectra provide information about intracellular molecular distributions, metabolism and chemical reactions, Raman microscopy has been widely utilized for bio-imaging and bio-functional analysis. By using metallic nanostructures, Raman scattering from molecules at the vicinity of a metal surface is significantly enhanced, which is called surface-enhanced Raman scattering (SERS). Due to localized field enhancement, the highly sensitive detection of molecules can be achieved and many attempts to apply it for biological and biomedical research have been made. In this article, we review the applications of metallic nanoparticles as SERS agents for molecular analysis and imaging of biological cells and tissues.
[Back to top]


Near-infrared spectroscopy in studies of brain oxygenation
Hideo Eda
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003747]

Near-infrared spectroscopy (NIRS) is widely used in studies of brain activity. In this article I explain the principles and theoretical limits of NIRS. I also discuss the need for the development of portable NIRS.
[Back to top]


Raman molecular imaging of cells and tissues: towards functional diagnostic imaging without labeling
Yoshinori Harada and Tetsuro Takamatsu
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003748]

Raman spectroscopy has long been used as a powerful tool for chemical composition analyses. Raman scattering light measurement has the advantage of being able to examine biomolecules in a nondestructive and non-labeling manner. However, molecular imaging of cells and tissues by using Raman microscopy had been hampered due to weak Raman signals and required long acquisition time. Recent advances in imaging devices, such as development of slit-scanning Raman microscopy, have enabled us to acquire high-resolution Raman images of biomedical samples. Thus, Raman molecular imaging now has wide application potential for in-situ functional analysis of biomolecules in living bodies, such as studies of intracellular drug pharmacokinetics and oxygen saturation of blood capillaries. There is a possibility that Raman scattering light measurement can be applied for functional diagnostic imaging of human bodies, taking advantage of its noninvasive merits in the future. This review briefly highlights recent topics in spontaneous Raman molecular imaging of cells and tissues.
[Back to top]


Coherent anti-Stokes Raman scattering microscopy for high speed non-staining biomolecular imaging
Mamoru Hashimoto,Takeo Minamikawa and Tsutomu Araki
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003749]

Vibrational microscopy (Raman microscopy and infrared microscopy), which observes molecular vibrations, gives us the information of molecular species without staining because the observed signals are originated from intrinsic molecules of a cell. However, infrared radiation is absorbed with water, and the long wavelength (3-10 _m) limits the spatial resolution to several micrometers. Spontaneous emission of Raman scattering is quite feeble, and the Raman scattering often overlaps with one-photon fluorescence from a specimen. Coherent anti-Stokes Raman scattering (CARS) microscopy, which is one of the nonlinear Raman microscopy, is a method to overcome those problems. In this review, present system of CARS microscopy, the methods of background rejection, and applications are introduced.
[Back to top]


Precise analysis of the autofluorescence characteristics of rat colon under UVA and violet light excitation
Keimei Nakano, Yoshinori Harada, Yoshihisa Yamaoka, Kiichiro Miyawaki, Katsuichi Imaizumi, Hideyuki Takaoka, Masaya Nakaoka, Naoki Wakabayashi, Toshikazu Yoshikawa and Tetsuro Takamatsu
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003750]

Purpose Tissue autofluorescence study is a promising means of endoscopic detection of colonic neoplasia, but the mechanism of autofluorescence eruption has still not been verified. The purpose of this study was to precisely analyze the autofluorescence characteristics of freshly prepared normal rat colon under UVA and violet light excitation.

Methods: Excised rat colons were studied by using multichannel spectrophotometry, spectroscopic imaging, confocal microscopy, combined two-photon excited fluorescence and second-harmonic generation (SHG) microscopy, and fluorescence lifetime imaging microscopy.

Results: Spectroscopic analysis of freshly prepared colon sections revealed that the mucosa and the submucosa showed strong autofluorescence under UVA and violet light excitation. The combined images of two-photon and SHG microscopy revealed that the mucosal epithelia are the important source of autofluorescence. Nicotinamide adenine dinucleotide seems to be one of the major substances involved in the autofluorescence of the mucosal layer on 365-nm light excitation. The autofluorescence spectra of the luminal surfaces were identical to those of the mucosa on cross-sectional examinations with 365-nm excitation. The main origin of autofluorescence of the luminal surface with 365-nm excitation is the epithelial cells in the mucosa without overlay of submucosal fluorescence.
[Back to top]


IR super-resolution microspectroscopy and its application to single cells
Makoto Sakai, Keiichi Inoue and Masaaki Fujii
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003751]

For many years, spatial resolution is the most critical problem in IR microspectroscopy. This is because the spatial resolution of a conventional infrared microscope is restricted by the diffraction limit, which is almost the same as the wavelength of IR light, ranging from 2.5 to 25 μm. In the recent years, we have developed two novel types of far-field IR super-resolution microscopes using 2-color laser spectroscopies, those are transient fluorescence detected IR (TFD-IR) spectroscopy and vibrational sum-frequency generation (VSFG) spectroscopy. In these ways, because both transient fluorescence and VSFG signal have a wavelength in the visible region, the image is observed at the resolution of visible light, which is about 10 times smaller than that of IR light (that is, IR super-resolution). By using these techniques, we can map the specific IR absorption band with sub-micrometer spatial resolution, visualization of the molecular structure and reaction dynamics in a non-uniform environment such as a cell becomes a possibility. In the present reviews, we introduce our novel IR super-resolution microspectroscopy and its application to single cells in detail.
[Back to top]


FRET Enhanced Fluorescent Nanodiamonds
Rafal Fudala, Sangram Rout, Badri P. Maliwal, T. W. Zerda, Ignacy Gryczynski, Eric Simanek, Julian Borejdo, Ryan Rich, Irina Akopova and Zygmunt Gryczynski
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003752]

Fluorescent nanodiamonds (FNDs) are one of the new and very promising biocompatible nanomaterials that can be used both as fluorescence imaging agent and a highly versatile platform for the controlled functionalization to target and deliver a wide spectrum of therapeutic agents. Among the remarkable fluorescence properties are excellent photostability, emission between 600-700nm, quantum yield of 1 and moderately long fluorescence lifetimes. However the low absorption cross section of fluorescent (N-V)- centers limits FNDs’ brightness. In this work we show that an approach based on the Forster resonance energy transfer (FRET) may significantly enhance the fluorescence signal observed from a single ND. We demonstrate that organic dyes (fluorophores) attached to the FND surface can efficiently transfer the excitation energy to (N-V)- centers. Multiple dyes positioned in close proximity to the ND facile surface may serve as harvesting antennas transferring excitation energy to the fluorescent centers. We propose that, with the help of some of the functional groups present on the FND surface, we can either directly link flurophores or use scalable dendrimer chemistry to position many organic dyes at a calibrated distance. Also, the remaining multiple functional groups will be still available for particle targeting and drug delivery. This opens new way for designing a new type of theranostics particles of ultra-high brightness, high photostability, specific targeting, and high capacity for drug delivery.
[Back to top]


Allosteric Inhibitors of Bcr-Abl: Towards Novel Myristate-Pocket Binders ?
Marco Radi, Silvia Schenone and Maurizio Botta
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003754]

Among the currently available options for the treatment of chronic myeloid leukemia (CML), ATP-competitive tyrosine kinases inhibitors (Gleevec®, Dasatinib® and Nilotinib®) represent one of the most promising therapeutic approaches developed in the last 10-15 years. However, the initial enthusiasm generated by the high response rate to these drugs has been dampened by the development of resistance. The T315I mutation at the gatekeeper residue is very frequent in advanced phases of the disease and is one of the main causes of resistance, disrupting important contact points between the inhibitors and the enzyme. Different strategies have been implemented to overcome this resistance, such as the recent development of more selective non-ATP competitive inhibitors targeting sites outside the ATP-binding cleft. Some of these allosteric inhibitors alter the kinase conformation, while others directly compete with the protein substrates. Another interesting family of allosteric inhibitors is represented by those compounds targeting the myristate-pocket of Bcr-Abl (myristate-pocket binders). The binding of these inhibitors blocks the Bcr-Abl kinase in the inactive conformation and provides an advantage in overcoming drug resistance due to kinase mutations. This review reports the last findings in the development of novel myristate-pocket binders of Bcr-Abl as promising anti-leukemia agents.
[Back to top]


Therapeutic Genes for Anti-HIV/AIDS Gene Therapy
chiara bovolenta, simona porcellini and luca alberici
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003755]

The multiple therapeutic approaches developed so far to cope HIV-1 infection, such as anti-retroviral drugs, germicides and several attempts of therapeutic vaccination have provided significant amelioration in terms of life-quality and survival rate of AIDS patients. Nevertheless, no approach has demonstrated efficacy in eradicating this lethal, if untreated, infection. The curative power of gene therapy has been proven for the treatment of monogenic immunodeficiensies, where permanent gene modification of host cells is sufficient to correct the defect for life-time. No doubt, a similar concept is not applicable for gene therapy of infectious immunodeficiensies as AIDS, where there is not a single gene to be corrected; rather engineered cells must gain immunotherapeutic or antiviral features to grant either short- or long-term efficacy mostly by acquisition of antiviral genes or payloads. Anti-HIV/AIDS gene therapy is one of the most promising strategy, although challenging, to eradicate HIV-1 infection. In fact, genetic modification of hematopoietic stem cells with one or multiple therapeutic genes is expected to originate blood cell progenies resistant to viral infection and thereby able to prevail on infected unprotected cells. Ultimately, protected cells will re-establish a functional immune system able to control HIV-1 replication. Sixty gene therapy clinical trials against AIDS employing different viral vectors and transgenes have been approved or are currently ongoing worldwide. This review will overview anti-HIV-1 infection gene therapy field evaluating strength and weakness of the transgenes and payloads used in the past and of those potentially exploitable in the future.
[Back to top]


Peptides as drugs: From screening to application
Ursula Dietrich, Ralf Dürr and Joachim Koch
[Purchase Article] [BSP/CPB/E-Pub/003756]

Peptides are ideally suited to mimic natural ligands and thereby function in an antagonistic or agonistic way. Furthermore they are able to physiologically disrupt functional complexes due to their small size and specific binding properties. Proteins form homo- or heteromeric (macro)molecular complexes and intricate networks by interacting with small molecules, peptides, nucleic acids or other proteins. On average, five interaction partners have been estimated for any given cellular protein, illustrating the complexity of the formed ‘interactomes’ and the impact of their investigation. Many protein-protein interactions are mediated by hot-spots, which comprise only a small part of the large binding interface but account for 80% of the binding energy. Thus, these hot spots provide an ‘Achilles heel’ for pharmaceutical interventions aiming at the disruption of functional protein-protein complexes.

Methods to select peptides for defined target structures include display technologies on phages, ribosomes or yeast, and the yeast-two-hybrid system. Once selected, these peptides can be optimized for their binding affinity using peptide arrays. Stabilization of biologically unstable peptides is achieved by the introduction of non-natural amino acids to form so-called peptidomimetics that are resistant to cellular proteases. Moreover, lipocalins and peptide aptamers represent scaffolded binding structures with unique binding characteristics and enhanced stability.

In case of extracellular targets, like cell surface receptors or pathogens in patients` plasma, peptide inhibitors have direct access. Addressing intracellular targets with peptides is more difficult since short hydrophilic peptides generally do not cross plasma membranes on their own. However, intracellular uptake of peptides can be achieved by coupling to carrier systems like liposomes or nanoparticles or upon fusion to a protein transduction domain. Alternatively, peptides may be expressed within cells after transduction with viral vectors in a gene therapy setting. This review will summarize the broad potential of peptides as drugs, with special emphasis on peptides which inhibit protein-protein interactions.
[Back to top]


Capsid Assembly as a Point of Intervention for Novel Anti-viral Therapeutics
Vishwanath R. Lingappa, Clarence R. Hurt and Edward Garvey
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003757]

In general, drug discovery in the therapeutic field of infectious disease has a stellar track record. And yet, subsets of pathogens, for example many classes of viruses other than HIV, HSV, influenza, and HCV have been poorly addressed. In addition, the development of resistance remains a specter of great concern for almost all current chemotherapy directed against infectious diseases, including viruses. Within the viral lifecycle, capsid assembly stands out as a step occurring in all viruses, which has not been the subject of extensive drug discovery programs.  Until recently, the common view of assembly was that all the necessary information for assembly was contained in the sequence of the viral protein, in other words, the capsid self-assembles. In the last few years, a body of data has opened new opportunities for antiviral pharmaceutical research. Evidence that host proteins may play catalytic or essential structural roles in viral capsid assembly suggests that these host proteins and their functions are novel targets for small molecule therapeutics. Here we review the current understanding of the capsid assembly process with emphasis on recent data that demonstrate the essential role of host proteins in capsid assembly.   Furthermore, this dependency of assembly on host factors appears quite sensitive to small molecule intervention.  Implications of this alternate mechanism of capsid assembly are also considered. For example, the dependency on host factors could impose a potent barrier to development of viral resistance to a host-targeted anti-capsid chemotherapeutic.  Finally, we give specific examples of the current state of drug discovery programs that have focused on therapeutic inhibition of host-assisted viral capsid assembly.
[Back to top]


Poly(ADP-ribosylation) and neoplastic transformation:Effect of PARP inhibitors
Francesca Donà, Ilaria Chiodi, Cristina Belgiovine, Tatiana Raineri, Roberta Ricotti, Chiara Mondello and A. Ivana Scovassi
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003758]

Poly(ADP-ribose) polymerases (PARPs) and poly(ADP-ribosylation) play essential roles in several biological processes, among which neoplastic transformation and telomere maintenance. In this paper, we review the poly(ADP-ribosylation) process together with the highly appealing use of PARP inhibitors for the treatment of cancer. In addition, we report our results concerning poly(ADP-ribosylation) in a cellular model system for neoplastic transformation developed in our laboratory. Here we show that PARP-1 and PARP-2 expression increases during neoplastic transformation, together with the basal levels of poly(ADP-ribosylation). Furthermore, we demonstrate a greater effect of the PARP inhibitor 3-aminobenzamide (3AB) on cellular viability in neoplastically transformed cells compared to normal fibroblasts and we show that prolonged 3AB administration to tumorigenic cells causes a decrease in telomere length. Taken together, our data support an active involvement of poly(ADP-ribosylation) in neoplastic transformation and telomere length maintenance and confirm the relevant role of poly(ADP-ribosylation) inhibition for the treatment of cancer.
[Back to top]


X-ray Crystallography as a tool for Mechanism-of-action Studies and Drug Discovery
Eric Ennifar
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003759]

Knowledge of three-dimensional structures of biological macromolecules is essential for a complete understanding of many biological processes. X-ray crystallography is the most widely used technique in structural biology and can provide highly detailed structures of proteins, nucleic acids or macromolecular complexes without any size limit. In the past decade, several recent advances in biological crystallography and automation of data collection and structure solution allowed extraordinary progresses and now more than 58 000 crystal structures have been deposited into the Protein Data Bank. This wealth of structural data significantly helped the elucidation of many biological processes and led to the development of new drugs. In this review we will show how of X-ray crystallography can provide insights into the mechanism of action of biological processes and can contribute to the rationale development of ligands through structure-based drug design.
[Back to top]


The power of enzyme kinetics in the drug development process
Alberta Samuele, Emmanuele Crespan, Anna Garbelli, Laura Bavagnoli and Giovanni Maga
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003760]

Drug development is a long and expensive process. It starts from the identification of a small molecule (hit compound) endowed with the ability to suppress a cellular or viral enzyme essential for the development of a given disease and proceeds through subsequent rounds of structural changes and optimization until the desired pharmacological properties are reached (lead compound). At any point of the hit-to-lead optimization process, it is of essence to monitor the behavior of the intermediate molecules with respect to their molecular targets. This involves precise mechanism of action studies as well as quantitative measurement of the performance of the compound against its target. Enzyme kinetic studies are thus an essential component of the drug development process. Relevant examples of the power of enzyme kinetics in the antiviral drug development process will be discussed in the context of anti-HIV chemotherapy.
[Back to top]


Simple and General Criterion for “In Silico” Screening of Candidate HIV Drugs
Nevena Veljkovic, Sanja Glisic, Jelena Prljic, Vladimir Perovic and Veljko Veljkovic
[FULL-TEXT INQUIRY] [BSP/CPB/E-Pub/003761]

Highly active antiretroviral therapy (HAART) dramatically has changed the course of HIV infection. Currently, this therapy involves the use of agents from at least two distinct classes of antivirals: a protease inhibitor in combination with two nucleoside/nucleotide reverse transcriptase inhibitors (N(t)RTIs), or a non-nucleoside reverse transcriptase inhibitor (NNRTI) in combination with NRTIs. Recently, the third family of antivirals started to be used clinically, with the advent of enfuvirtide, the first fusion inhibitor. This broad spectrum of anti-HIV agents recently was extended with compounds inhibiting HIV integrase and vital entry. But these advances did not come without a cost: there were the short- and long-term drug toxicities, emergence of drug resistance, and persistence of viral reservoirs. For these reasons, there is a pressing need for novel anti-HIV drugs, particularly those that have a novel action mechanism, as these might be less likely to show cross-resistance with current therapies. The field of bioinformatics has become a major part of the drug discovery pipeline playing a key role in improvement and acceleration of the time and money consuming process of the drug development. Here we review the application of the EIIP/AQVN (Electron-Ion Interaction Potential, EIIP; Average Quasi Valence Number, AQVN) bioinformatics concept in the development of new anti-HIV drugs and propose a simple theoretical criterion for a virtual screening of molecular libraries for promising lead anti-HIV compounds and refinement of selected lead compounds in order to increase their biological activity.
[Back to top]