Guava Publications

At Sigma-Aldrich, we aim to bring innovative solutions closer to you. The publications below show how our products play a part in research.


 October 2017


A cellular threshold for active ERK1/2 levels determines Raf/MEK/ERK-mediated growth arrest versus death responses
Original Research Article
Cellular Signalling, Available online 3 October 2017, Pages
Seung-Keun Hong, Pui-Kei Wu, Jong-In Park


 September 2017


Effects of growth phase and nitrogen limitation on biochemical composition of two strains of Tisochrysis lutea
Original Research Article
Algal Research, Volume 27, November 2017, Pages 177-189

Fiz da Costa, Fabienne Le Grand, Claudie Quéré, Gaël Bougaran, Jean Paul Cadoret, René Robert, Philippe Soudant
Differential effects of FXR or TGR5 activation in cholangiocarcinoma progression
Original Research Article
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, Available online 13 September 2017, Pages

O. Erice, I. Labiano, A. Arbelaiz, A. Santos-Laso, P. Munoz-Garrido, R. Jimenez-Agüero, P. Olaizola, A. Caro-Maldonado, N. Martín-Martín, A. Carracedo, E. Lozano, J.J. Marin, C.J. O'Rourke, J.B. Andersen, J. Llop, V. Gómez-Vallejo, D. Padro, A. Martin, M. Marzioni, L. Adorini, M. Trauner, L. Bujanda, M.J. Perugorria, J.M. Banales
In Vivo Selection of a Computationally Designed SCHEMA AAV Library Yields a Novel Variant for Infection of Adult Neural Stem Cells in the Subventricular Zone
Original Research Article
Molecular Therapy, Available online 8 September 2017, Pages

David S. Ojala, Sabrina Sun, Jorge L. Santiago-Ortiz, Mikhail G. Shapiro, Philip A. Romero, David V. Schaffer
In vitro antitumor activity of novel benzimidazole-based Cu(II) complexes
Original Research Article
Bioorganic & Medicinal Chemistry, Available online 5 September 2017, Pages

Jiyong Hu, Yan Guo, Jin'an Zhao, Junshuai Zhang

Graphical abstract


Twinfilin 1 enhances milk bio-synthesis and proliferation of bovine mammary epithelial cells via the mTOR signaling pathway
Biochemical and Biophysical Research Communications, Available online 31 August 2017, Pages
Lu Li, Lijie Liu, Bo Qu, Xueying Li, Xuejun Gao, Minghui Zhang
A PEGylated hyaluronic acid conjugate for targeted cancer immunotherapy
Original Research Article
Journal of Controlled Release, Available online 26 August 2017, Pages

Jung Min Shin, Se Jin Oh, Seunglee Kwon, V.G. Deepagan, Minchang Lee, Seok Ho Song, Hyo-Jung Lee, Suyeon Kim, Kwon-Ho Song, Tae Woo Kim, Jae Hyung Park

Graphical abstract



 August 2017


DNA-loaded cationic liposomes efficiently function as a vaccine against malarial proteins   
Original Research Article
Molecular Therapy - Methods & Clinical Development, Available online 23 August 2017, Pages
Wesley L. Fotoran, Rachele Santangelo, Beatriz N.M. de Miranda, Darrell J. Irvine, Gerhard Wunderlich

Graphical Abstract

Extracellular matrix remodeling and matrix metalloproteinase inhibition in visceral adipose during weight cycling in mice
Original Research Article
Experimental Cell Research, Available online 19 August 2017, Pages
Cíntia Rabelo e Paiva Caria, Érica Martins Ferreira Gotardo, Paola Souza Santos, Simone Coghetto Acedo, Thainá Rodrigues de Morais, Marcelo Lima Ribeiro, Alessandra Gambero
Baculovirus-Induced Recombinant Protein Expression in Human Mesenchymal Stromal Stem Cells: A Promoter Study   
New Biotechnology, Available online 19 August 2017, Pages
Gundula Sprick, Tobias Weidner, Denise Salzig, Peter Czermak
Generation of iPSC line from desmin-related cardiomyopathy patient carrying splice site mutation of DES gene   
Stem Cell Research, Available online 19 August 2017, Pages
Aleksandr Khudiakov, Daria Kostina, Anna Zlotina, Tatiana Nikulina, Alexey Sergushichev, Alexandra Gudkova, Alexey Tomilin, Anna Malashicheva, Anna Kostareva
Generation of iPSC line from patient with arrhythmogenic right ventricular cardiomyopathy carrying mutations in PKP2 gene
Stem Cell Research, Available online 18 August 2017, Pages

Aleksandr Khudiakov, Daria Kostina, Anna Zlotina, Natalia Yany, Alexey Sergushichev, Tatiana Pervunina, Alexey Tomilin, Anna Kostareva, Anna Malashicheva
Synergistic antitumor effects of combination treatment with metronomic doxorubicin and VEGF-targeting RNAi nanoparticles
Original Research Article
Journal of Controlled Release, Available online 16 August 2017, Pages
Gijung Kwak, Sung Duk Jo, Dongkyu Kim, Hyosuk Kim, Myung Goo Kim, Kwangmeyung Kim, Ick Chan Kwon, Sun Hwa Kim

Graphical abstract

Improved therapeutic efficacy of mammalian expressed-recombinant interferon gamma against ovarian cancer cells   
Original Research Article
Experimental Cell Research, Available online 10 August 2017, Pages
Ali Razaghi, Carina Villacrés, Vincent Jung, Narges Mashkour, Michael Butler, Leigh Owens, Kirsten Heimann

Graphical abstract

A novel RIPK1 inhibitor that prevents retinal degeneration in a rat glaucoma model   
Original Research Article
Experimental Cell Research, Available online 9 August 2017, Pages
Yun-Ju Do, Jee-Won Sul, Ki-Hong Jang, Nam Sook Kang, Young-Hoon Kim, Young-Gwan Kim, Eunhee Kim
Derivation of a disease-specific human induced pluripotent stem cell line from a biliary atresia patient   
Stem Cell Research, Available online 8 August 2017, Pages

Lipeng Tian, Lindsey Eldridge, Pooja Chaudhari, Linyi Zhang, Robert A. Anders, Kathleen B. Schwarz, Zhaohui Ye, Yoon-Young Jang
Biochemical responses to cadmium exposure in Oncorhynchus mykiss erythrocytes   
Original Research Article
Ecotoxicology and Environmental Safety, Volume 145, November 2017, Pages 476-482
Patrick Orlando, Sonia Silvestri, Enea Ferlizza, Giulia Andreani, Emilio Carpenè, Giancarlo Falcioni, Luca Tiano, Gloria Isani
Platelet Toll-like receptor (TLR) expression and TLR-mediated platelet activation in acute myocardial infarction   
Original Research Article
Thrombosis Research, Volume 158, October 2017, Pages 8-15
Kathryn E. Hally, Anne C. La Flamme, Peter D. Larsen, Scott A. Harding
Significant inhibition of Tembusu virus envelope and NS5 gene using an adenovirus-mediated short hairpin RNA delivery system   
Original Research Article
Infection, Genetics and Evolution, Available online 3 August 2017, Pages
Hongzhi Wang, Qiang Feng, Lei Wei, Liling Zhuo, Hao Chen, Youxiang Diao, Yi Tang
[Ru(pipe)(dppb)(bipy)]PF6: A novel ruthenium complex that effectively inhibits ERK activation and cyclin D1 expression in A549 cells   
Original Research Article
Toxicology in Vitro, Available online 1 August 2017, Pages
Guilherme A. Ferreira-Silva, Marina M. Ortega, Marco A. Banionis, Graciana Y. Garavelli, Felipe T. Martins, Julia S.M. Dias, Cláudio Viegas, Jaqueline C. de Oliveira, Fabio B. do Nascimento, Antonio C. Doriguetto, Marilia I.F. Barbosa, Marisa Ionta


 July 2017


Pyruvate kinase M1 interacts with A-Raf and inhibits endoplasmic reticulum stress-induced apoptosis by activating MEK1/ERK pathway in mouse insulinoma cells   
Original Research Article
Cellular Signalling, Volume 38, October 2017, Pages 212-222
Yuta Horiuchi, Daiki Nakatsu, Fumi Kano, Masayuki Murata 

Graphical abstract

Long Non-coding RNA–mRNA Correlation Analysis Reveals the Potential Role of HOTAIR in Pathogenesis of Sporadic Thoracic Aortic Aneurysm   
Original Research Article
European Journal of Vascular and Endovascular Surgery, Available online 27 July 2017, Pages
X. Guo, Q. Chang, H. Pei, X. Sun, X. Qian, C. Tian, H. Lin
Pro-inflammatory chemokines and cytokines dominate the blister fluid molecular signature in epidermolysis bullosa patients and affect leukocyte and stem cell migration   
Original Research Article
Journal of Investigative Dermatology, Available online 20 July 2017, Pages
Vitali Alexeev, Julio Cesar Salas-Alanis, Francis Palisson, Lila Mukhtarzada, Giulio Fortuna, Jouni Uitto, Andy South, Olga Igoucheva
Migration ability and Toll-like receptor expression of human mesenchymal stem cells improves significantly after three-dimensional culture   
Biochemical and Biophysical Research Communications, Available online 19 July 2017, Pages
Panpan Zhou, Zilin Liu, Xue Li, Bing Zhang, Xiaoyuan Wang, Jing Lan, Qing Shi, Dong Li, Xiuli Ju
Tyrosine kinase inhibitors as modulators of trastuzumab-mediated antibody-dependent cell-mediated cytotoxicity in breast cancer cell lines   
Original Research Article
Cellular Immunology, Available online 15 July 2017, Pages
Denis M. Collins, Kathy Gately, Clare Hughes, Connla Edwards, Anthony Davies, Stephen F. Madden, Kenneth J. O'Byrne, Norma O'Donovan, John Crown


 May 2017


Thiosemicarbazones and 4-thiazolidinones indole-based derivatives: Synthesis, evaluation of antiproliferative activity, cell death mechanisms and topoisomerase inhibition assay   
Original Research Article
European Journal of Medicinal Chemistry, Volume 136, 18 August 2017, Pages 305-314
Jamerson Ferreira de Oliveira, Talitha Santos Lima, Débora Barbosa Vendramini-Costa, Sybelle Christianne Batista de Lacerda Pedrosa, Elizabeth Almeida Lafayette, Rosali Maria Ferreira da Silva, Sinara Monica Vitalino de Almeida, Ricardo Olímpio de Moura, Ana Lúcia Tasca Gois Ruiz, João Ernesto de Carvalho, Maria do Carmo Alves de Lima

Graphical abstract


Upregulation of the long non-coding RNA SNHG1 predicts poor prognosis, promotes cell proliferation and invasion, and reduces apoptosis in glioma   
Original Research Article
Biomedicine & Pharmacotherapy, Volume 91, July 2017, Pages 906-911
Qiang Wang, Qing Li, Peng Zhou, Danni Deng, Lian Xue, Naiyuan Shao, Ya Peng, Feng Zhi

Graphical abstract


Molecular Diagnosis of Mosaic Overgrowth Syndromes Using a Custom-Designed Next-Generation Sequencing Panel 
Original Research Article
The Journal of Molecular Diagnostics, Available online 11 May 2017, Pages
Fengqi Chang, Liu Liu, Erica Fang, Guangcheng Zhang, Tiansheng Chen, Kajia Cao, Yanchun Li, Marilyn M. Li
Apoptosis induction by 7-chloroquinoline-1,2,3-triazoyl carboxamides in triple negative breast cancer cells   
Original Research Article
Biomedicine & Pharmacotherapy, Volume 91, July 2017, Pages 510-516
Karine Rech Begnini, Wladimir R. Duarte, Liziane Pereira da Silva, Julieti H. Buss, Bruna S. Goldani, Mariana Fronza, Natália Vieira Segatto, Diego Alves, Lucielli Savegnago, Fabiana Kömmling Seixas, Tiago Collares
Cell culture media supplemented with raffinose reproducibly enhances high mannose glycan formation   
Original Research Article
Journal of Biotechnology, Available online 29 April 2017, Pages
David Brühlmann, Anais Muhr, Rebecca Parker, Thomas Vuillemin, Blanka Bucsella, Franka Kalman, Serena Torre, Fabio La Neve, Antonio Lembo, Tobias Haas, Markus Sauer, Jonathan Souquet, Hervé Broly, Jürgen Hemberger, Martin Jordan
Glycolipids from spinach suppress LPS-induced vascular inflammation through eNOS and NK-κB signaling   
Original Research Article
Biomedicine & Pharmacotherapy, Volume 91, July 2017, Pages 111-120
Masakazu Ishii, Tatsuo Nakahara, Daisuke Araho, Juri Murakami, Masahiro Nishimura
Prediction of the fertility of stallion frozen-thawed semen using a combination of computer-assisted motility analysis, microscopical observation and flow cytometry   
Original Research Article
Theriogenology, Available online 27 April 2017, Pages
I. Barrier Battut, A. Kempfer, N. Lemasson, L. Chevrier, S. Camugli
Treatment schedule and estrogen receptor-status influence acquisition of doxorubicin resistance in breast cancer cells
Original Research Article
European Journal of Pharmaceutical Sciences, Available online 26 April 2017, Pages
Logeswari Ponnusamy, Prathap Kumar S. Mahalingaiah, Kamaleshwar P. Singh
Copper(II) complexes inducing apoptosis in cancer cells, and demonstrating DNA and HSA interactions   
Original Research Article
Polyhedron, Available online 26 April 2017, Pages
Jiyong Hu, Chunli Liao, Yan Guo, Fan Yang, Wei Sang, Jin'an Zhao


 April 2017

Facile encapsulation of hydroxycamptothecin nanocrystals into zein-based nanocomplexes for active targeting drug delivery and cell imaging   
Original Research Article
Acta Biomaterialia, Available online 19 April 2017, Pages
Hongdi Wang, Wei Zhu, Yunna Huang, Zhixian Li, Yanbin Jiang, Qiuling Xie

Graphical abstract  


Susceptibility of stallion spermatozoa to different oxidative challenges: role of seminal plasma   
Original Research Article
Journal of Equine Veterinary Science, Available online 18 April 2017, Pages
G.K.V. Kawai, J.R.C. Gurgel, J.D.A. Losano, A. Dalmazzo, C.C. Rocha, R.H. Tsunoda, P.A.A. Góes, B.R. Rui, D.S.R. Angrimani, M.E.O.A. Assumpção, C.M. Mendes, V.H. Barnabe, M. Nichi 
Peptidoglycan accelerates granulopoiesis through a TLR2- and MyD88-dependent pathway   
Biochemical and Biophysical Research Communications, Available online 15 April 2017, Pages
Masaya Takehara, Soshi Seike, Teruhisa Takagishi, Keiko Kobayashi, Masahiro Nagahama 
First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri   
Original Research Article
Microbial Pathogenesis, Volume 107, June 2017, Pages 219-224
Junyan Liu, Lin Li, Bing Li, Brian M. Peters, Zhenbo Xu, Mark E. Shirtliff 
CuS Nanoagents for Photodynamic and Photothermal Therapies: Phenomena and Possible Mechanisms   
Original Research Article
Photodiagnosis and Photodynamic Therapy, Available online 5 April 2017, Pages
Lihua Li, Leila H. Rashidi, Mengyu Yao, Lun Ma, Lingling Chen, Junying Zhang, Yu Zhang, Wei Chen 
First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri   
Original Research Article
Microbial Pathogenesis, Volume 107, June 2017, Pages 219-224
Junyan Liu, Lin Li, Bing Li, Brian M. Peters, Zhenbo Xu, Mark E. Shirtliff 
CuS Nanoagents for Photodynamic and Photothermal Therapies: Phenomena and Possible Mechanisms   
Original Research Article
Photodiagnosis and Photodynamic Therapy, Available online 5 April 2017, Pages
Lihua Li, Leila H. Rashidi, Mengyu Yao, Lun Ma, Lingling Chen, Junying Zhang, Yu Zhang, Wei Chen 


 March 2017


Early-stage autophagy protects nucleus pulposus cells from glucose deprivation-induced degeneration via the p-eIF2α/ATF4 pathway   
Original Research Article
Biomedicine & Pharmacotherapy, Volume 89, May 2017, Pages 529-535
Hongze Chang, Feng Cai, Yan Zhang, Mintao Xue, Liang Liu, Anli Yang, Xiaodong Liu
Linking of mPGES-1 and iNOS activates stem-like phenotype in EGFR-driven epithelial tumor cells   
Original Research Article
Nitric Oxide, Available online 1 March 2017, Pages
Erika Terzuoli, Federica Finetti, Filomena Costanza, Antonio Giachetti, Marina Ziche, Sandra Donnini

Graphical abstract  


miR-600 Acts as a Bimodal Switch that Regulates Breast Cancer Stem Cell Fate through WNT Signaling   
Original Research Article
Cell Reports, Volume 18, Issue 9, 28 February 2017, Pages 2256-2268
Rita El Helou, Guillaume Pinna, Olivier Cabaud, Julien Wicinski, Ricky Bhajun, Laurent Guyon, Claire Rioualen, Pascal Finetti, Abigaelle Gros, Bernard Mari, Pascal Barbry, Francois Bertucci, Ghislain Bidaut, Annick Harel-Bellan, Daniel Birnbaum, Emmanuelle Charafe-Jauffret, Christophe Ginestier

Graphical Abstract  

Induction of Chromosome Instability by Activation of Yes Associated Protein and Forkhead box M1 in Liver Cancer
Original Research Article
Gastroenterology, Available online 27 February 2017, Pages
Sofia M.E. Weiler, Federico Pinna, Thomas Wolf, Teresa Lutz, Aman Geldiyev, Carsten Sticht, Maria Knaub, Stefan Thomann, Michaela Bissinger, Shan Wan, Stephanie Rössler, Diana Becker, Norbert Gretz, Hauke Lang, Frank Bergmann, Vladimir Ustiyan, Tatiana V. Kalin, Stephan Singer, Ju-Seog Lee, Jens U. Marquardt, Peter Schirmacher, Vladimir V. Kalinichenko, Kai Breuhahn
Induction of Chromosome Instability by Activation of Yes Associated Protein and Forkhead box M1 in Liver Cancer
Original Research Article
Gastroenterology, Available online 27 February 2017, Pages
Sofia M.E. Weiler, Federico Pinna, Thomas Wolf, Teresa Lutz, Aman Geldiyev, Carsten Sticht, Maria Knaub, Stefan Thomann, Michaela Bissinger, Shan Wan, Stephanie Rössler, Diana Becker, Norbert Gretz, Hauke Lang, Frank Bergmann, Vladimir Ustiyan, Tatiana V. Kalin, Stephan Singer, Ju-Seog Lee, Jens U. Marquardt, Peter Schirmacher, Vladimir V. Kalinichenko, Kai Breuhahn
Effect of diterpenoid kaurenoic acid on genotoxicity and cell cycle progression in gastric cancer cell lines   
Original Research Article
Biomedicine & Pharmacotherapy, Volume 89, May 2017, Pages 772-780
Plínio Cerqueira dos Santos Cardoso, Carlos Alberto Machado da Rocha, Mariana Ferreira Leal, Marcelo de Oliveira Bahia, Diego Di Felipe Ávila Alcântara, Raquel Alves dos Santos, Natália dos Santos Gonçalves, Sérgio Ricardo Ambrósio, Bruno Coêlho Cavalcanti, Caroline Aquino Moreira-Nunes, Claudia do Ó Pessoa, Rommel Mário Rodríguez Burbano
Interactions between tafenoquine and artemisinin-combination therapy partner drug in asexual and sexual stage Plasmodium falciparum   
Original Research Article
International Journal for Parasitology: Drugs and Drug Resistance, Volume 7, Issue 2, August 2017, Pages 131-137
Karen Kemirembe, Mynthia Cabrera, Liwang Cui

Graphical abstract  


Sensitivity of eastern oyster (Crassostrea virginica) spermatozoa and oocytes to dispersed oil: Cellular responses and impacts on fertilization and embryogenesis   
Original Research Article
Environmental Pollution, Available online 24 March 2017, Pages
J. Vignier, A.K. Volety, A. Rolton, N. Le Goïc, F.-L.E. Chu, R. Robert, P. Soudant

Graphical abstract  

Microcarrier choice and bead-to-bead transfer for human mesenchymal stem cells in serum-containing and chemically defined media   
Review Article
Process Biochemistry, Available online 22 March 2017, Pages
Jasmin Leber, Jan Barekzai, Miriam Blumenstock, Björn Pospisil, Denise Salzig, Peter Czermak

Graphical abstract  


Anticancer copper(II) phosphorus dendrimers are potent proapoptotic Bax activators   
Original Research Article
European Journal of Medicinal Chemistry, Available online 21 March 2017, Pages
Serge Mignani, Nabil El Brahmi, Laure Eloy, Joel Poupon, Valérie Nicolas, Anke Steinmetz, Said El Kazzouli, Mosto M. Bousmina, Mireille Blanchard-Desce, Anne-Marie Caminade, Jean-Pierre Majoral, Thierry Cresteil

Graphical abstract 

Multivalent phosphorus dendrimers are antiproliferative agents displaying distinct mode of action: activation of caspase-3 for the plain dendrimer and stimulation of Bax translocation and AIF release for the Cu-complexed dendrimer.Image 1  


Cytoplasmic droplet acting as a mitochondrial modulator during sperm maturation in dogs   
Original Research Article
Animal Reproduction Science, Available online 30 March 2017, Pages
D.S.R. Angrimani, J.D.A. Losano, C.F. Lucio, G.A.L. Veiga, F.C. Landim, M. Nichi, C.I. Vannucchi
Notch signaling pathway promotes the development of ovine ovarian follicular granulosa cells   
Original Research Article
Animal Reproduction Science, Available online 28 March 2017, Pages
Jiongjie Jing, Xiaolong Jiang, Jianwei Chen, Xiaolei Yao, Miaomiao Zhao, Pengfei Li, Yangyang Pan, Youshe Ren, Wenzhong Liu, Lihua Lyu
Shelf life of diluted human interferon-α for veterinary clinical use   
Original Research Article
Biologicals, Available online 27 March 2017, Pages
Massimo Amadori, Giovanni Fiorentini


 February 2017


Immunological, clinical, haematological and oxidative responses to long distance transportation in horses
Original Research Article
Research in Veterinary Science, Volume 115, December 2017, Pages 78-87

Barbara Padalino, Sharanne Lee Raidal, Nicole Carter, Pietro Celi, Gary Muscatello, Leo Jeffcott, Kumudika de Silva
Evaluation of changes in promoters, use of UCOES and chain order to improve the antibody production in CHO cells
Original Research Article
Protein Expression and Purification, Available online 2 February 2017, Pages

Maria del Refugio Rocha-Pizaña, Guadalupe Ascencio-Favela, Brenda Maribell Soto-García, M.L. Martinez-Fierro, Moisés Alvarez Mario
Silica nanoparticles induced intrinsic apoptosis in neuroblastoma SH-SY5Y cells via CytC/Apaf-1 pathway
Original Research Article
Environmental Toxicology and Pharmacology, Available online 30 January 2017, Pages

Yanyan Yang, Yongbo Yu, Jiahui Wang, Yanbo Li, Yang Li, Jia Wei, Tong Zheng, Minghua Jin, Zhiwei Sun


 November 2016


Structural changes of Arthrospira sp. after low energy sonication treatment for microalgae harvesting: Elucidating key parameters to detect the rupture of gas vesicles

Bioresource Technology

Martí Lecina, Benjamin Sanchez, Carles Solà, Jordi Prat, Mònica Roldán, Mariona Hernández, Ramon Bragós, Carlos J. Paredes, Jordi J. Cairó

The buoyancy suppression by low energy sonication (LES) treatment (0.8 W·mL−1, 20 kHz, 10 s) has recently been proposed as an initial harvesting step for Arthrospira sp. This paper aims to describe the structural changes in Arthrospira sp. after LES treatment and to present how these structural changes affect the results obtained by different analytical techniques. Transmission electron microscopy (TEM) micrographs of trichomes evidenced the gas vesicles rupture but also revealed a rearrangement of thylakoids and more visible phycobilisomes were observed. Differences between treated and untreated samples were detected by confocal microscopy, flow cytometry and optical microscopy but not by electrical impedance spectroscopy (EIS). After LES treatment, 2-fold increase in autofluorescence at 610/660 nm was measured (phycocyanin/allophycocyanin emission wavelengths) and a ten-fold decrease in side scatter light intensity (due to a reduction of trichome’s inner complexity). This was further confirmed by optical microscopy showing changes on trichomes appearance (from wrinkled to smooth).

Toll-like receptor 9 expression and activation in acute coronary syndrome patients on dual anti-platelet therapy

Thrombosis Research

Kathryn E Hally, Anne C La Flamme, Peter D Larsen, Scott A Harding

The Toll-like receptor 9 (TLR9) pathway can activate platelets but its role in acute coronary syndromes (ACS) is unknown. This study examined TLR9 expression and platelet activation in response to ODN2006, a TLR9 agonist, in healthy subjects and in ACS subjects treated with dual anti-platelet therapy (DAPT).


TLR9 expression was examined in both resting and thrombin receptor activator peptide (TRAP)-activated platelets (1 and 10 μM) from healthy and ACS subjects by flow cytometry. In both cohorts, ODN2006-mediated platelet activation (5 μM) was examined in whole blood (WB) and platelet-rich plasma (PRP) using cell-surface CD62p and CD63 expression by flow cytometry.


Baseline TLR9 expression was significantly greater in ACS subjects compared to healthy subjects (p < 0.01). Following TRAP activation, TLR9 expression increased dose-dependently in healthy subjects. However, no difference in TLR9 expression was seen in ACS platelets following TRAP activation. ODN2006 treatment resulted in significant increases in cell-surface expression of CD62p and CD63 in both WB (all p < 0.001) and PRP (all p < 0.001) in comparison to unstimulated platelets in healthy subjects. Despite DAPT, ODN2006 treatment produced significant increases in both activation markers in the ACS cohort across WB and PRP (all p < 0.0001). Elevated baseline expression of TLR9 in ACS platelets may indicate increased sensitivity to TLR9 agonists and contribute to increased platelet activation in these patients. Furthermore, ODN2006 stimulation can activate platelets in ACS subjects despite treatment with DAPT.


This study demonstrates TLR9 expression and activation to be of potential therapeutic importance in ASC patients.

Macrophage integrins modulate response to ultra-high molecular weight polyethylene particles and direct particle-induced osteolysis  


Toral D. Zaveri, Natalia V. Dolgova, Jamal S. Lewis, Kiri Hamaker, Michael J. Clare-Salzler, Benjamin G. Keselowsky

Aseptic loosening due to peri-prosthetic osteolysis is one of the primary causes for failure of artificial joint replacements. Implant-derived wear particles, often ultra-high molecular weight polyethylene (UHMWPE) microparticles, initiate an inflammatory cascade upon phagocytosis by macrophages, which leads to osteoclast recruitment and activation, ultimately resulting in osteolysis. Investigation into integrin receptors, involved in cellular interactions with biomaterial-adsorbed adhesive proteins, is of interest to understand and modulate inflammatory processes. In this work, we investigate the role of macrophage integrins Mac-1 and RGD-binding integrins in response to UHMWPE wear particles. Using integrin knockout mice as well as integrin blocking techniques, reduction in macrophage phagocytosis and inflammatory cytokine secretion is demonstrated when these receptors are either absent or blocked. Along this line, various opsonizing proteins are shown to differentially modulate microparticle uptake and macrophage secretion of inflammatory cytokines. Furthermore, using a calvarial osteolysis model it is demonstrated that both Mac-1 integrin and RGD-binding integrins modulate the particle induced osteolysis response to UHMWPE microparticles, with a 40% decrease in the area of osteolysis by the absence or blocking of these integrins, in vivo. Altogether, these findings indicate Mac-1 and RGD-binding integrins are involved in macrophage-directed inflammatory responses to UHMWPE and may serve as therapeutic targets to mitigate wear particle induced peri-prosthetic osteolysis for improved performance of implanted joints.

Casearin D inhibits ERK phosphorylation and induces downregulation of cyclin D1 in HepG2 cells   

Toxicology in Vitro

Guilherme Álvaro Ferreira-Silva, Carla Carolina Lopes Lages, Patricia Sartorelli, Flávia Rie Hasegawa, Marisi Gomes Soares, Marisa Ionta

Cancer is a public health problem which represents the second cause of death in the world. In this framework, it is necessary to identify novel compounds with antineoplastic potential. Plants are an important source for discovering novel compounds with pharmacological potential. In this study, we aimed to investigate the antiproliferative potential of isolated compounds from Casearia sylvestris on tumor cell lines. Crude extract effectively reduced cell viability of 4 tumor cell lines (HepG2, A549, U251-MG, and HT-144) after 48 h treatment. HepG2 and HT-144 were the most responsive cells. Three fractions (aqueous ethanol, n-hexane and ethyl acetate) were tested against HepG2 and HT-144 cells and we observed that compounds with antiproliferative activity were concentrated in n-hexane and ethyl acetate fractions. The casearins A, G and J were isolated from n-hexane fraction, while casearin D was obtained from ethyl acetate fraction. We demonstrated that casearin D significantly inhibited the clonogenic capacity of HepG2 cells after 24 h exposure indicating its antiproliferative activity. In addition, G1/S transition cell cycle arrest in HepG2 cells was also observed. These effects are related, at least in part, to ability of the casearin D in reducing ERK phosphorylation and cyclin D1 expression levels.

Never deem lightly the “less harmful” low-molecular-weight PAH, NPAH, and OPAH — Disturbance of the immune response at real environmental levels   


Cui Wang, Jinhuan Yang, Linwensi Zhu, Lu Yan, Dezhao Lu, Quan Zhang, Meirong Zhao, Zhuoyu Li

The upcoming energy structure optimization and the implementation of strict emissions control will effectively alleviated the pollution of high-molecular-weight (HMW) polycyclic aromatic hydrocarbons (PAHs) in the atmosphere. Compared to HMW PAHs, the immune response to low-molecular-weight (LMW) PAHs is recognized as “less harmful”, despite the high proportions of these substances. The present study intends to investigate the effects of several of the most abundant LMW PAHs on macrophages RAW264.7 at environmentally relevant doses. The data assembled herein showed that Fluoranthene (Fluo, PAH) formed a π-π interaction with the Phe12 residue of AhR while inhibiting the transcription of CYP1A1 and CYP1B1, and ultimately induced the inflammatory cytokines in RAW264.7. The 1-Nitropyrene (1-Nitro, NPAH) formed both a π-π interaction and a hydrogen bond with AhR, stimulated CYP1A1transcription, while suppressed the cytokine levels. Additionally, the inflammation potency caused by TPAHs was highly correlated with the cytotoxic potency rather than the oxidative stress potency. When stimulated by LPS, the transcription of IL-6 was inhibited by Fluo, and 1-Nitro suppressed both IL-6 and TNFα transcription. Furthermore, only 1-Nitro gave a significant inhibition on phagocytosis. The effects of 9-Fluorenone (9-Fluo, OPAH) on macrophages remained insignificant throughout the study since the low affinity for AhR, which resulted in low cytotoxicity. Collectively, this study suggested that LMW PAHs tended to cause mild inflammation when they bind without activating AhR. During infection, AhR ligands caused immunosuppression and this potency for TPAHs may be higher in AhR activator than that in AhR inactivator.

Immune priming and portal of entry effectors improve response to vibrio infection in a resistant population of the European abalone   

Fish & Shellfish Immunology

Bruno Dubief, Flavia L.D. Nunes, Olivier Basuyaux, Christine Paillard

Since 1997, populations of the European abalone Haliotis tuberculata suffer mass mortalities attributed to the bacterium Vibrio harveyi. These mortalities occur at the spawning season, when the abalone immune system is depressed, and when temperatures exceed 17 °C, leading to favorable conditions for V. harveyi proliferation. In order to identify mechanisms of disease resistance, experimental successive infections were carried out on two geographically distinct Brittany populations: one that has suffered recurrent mortalities (Saint-Malo) and one that has not been impacted by the disease (Molène). Furthermore, abalone surviving these two successive bacterial challenges and uninfected abalone were used for several post-infection analyses. The Saint-Malo population was found to be resistant to V. harveyi infection, with a survival rate of 95% compared to 51% for Molène. While in vitro quantification of phagocytosis by flow cytometry showed strong inhibition following the first infection, no inhibition of phagocytosis was observed following the second infection for Saint-Malo, suggesting an immune priming effect. Moreover, assays of phagocytosis of GFP-labelled V. harveyi performed two months post-infection show an inhibition of phagocytosis by extracellular products of V. harveyi for uninfected abalone, while no effect was observed for previously infected abalone from Saint-Malo, suggesting that the effects of immune priming may last upwards of two months. Detection of V. harveyi by qPCR showed that a significantly greater number of abalone from the susceptible population were positive for V. harveyi in the gills, indicating that portal of entry effectors may play a role in resistance to the disease. Collectively, these results suggest a potential synergistic effect of gills and hemolymph in the resistance of H. tuberculata against V. harveyi with an important involvement of the gills, the portal of entry of the bacteria.

Celecoxib-loaded PEA microspheres as an auto regulatory drug-delivery system after intra-articular injection   

Journal of Controlled Release

Maarten Janssen, Ufuk Tan Timur, Nina Woike, Tim JM Welting, Guy Draaisma, Marion Gijbels, Lodewijk van Rhijn, George Mihov, Jens Thies, Pieter Emans

In this study, we investigated the potential of celecoxib-loaded polyester amide (PEA) microspheres as an auto-regulating drug delivery system for the treatment of pain associated with knee osteoarthritis (OA). Celecoxib release from PEA microspheres and inflammation responsive release of a small molecule from PEA was investigated in vitro. Inflammation responsive release of a small molecule from PEA was observed when PEA was exposed to cell lysates obtained from a neutrophil-like Hl-60 cell line. Following a short initial burst release of ~ 15% of the total drug load in the first days, celecoxib was slowly released throughout a period of > 80 days. To investigate biocompatibility and degradation behavior in vivo, celecoxib-loaded PEA microspheres were injected in OA-induced (ACLT + pMMx) or contralateral healthy knee joints of male Lewis rats. Bioactivity of celecoxib from loaded PEA microspheres was confirmed by PGE2 measurements in total rat knee homogenates. Intra-articular biocompatibility was demonstrated histologically, where no cartilage damage or synovial thickening and necrosis were observed after intra-articular injections with PEA microspheres. Degradation of PEA microspheres was significantly higher in OA induced knees compared to contralateral healthy knee joints, while loading the PEA microspheres with celecoxib significantly inhibited degradation, indicating a drug delivery system with auto regulatory behavior. In conclusion, this study suggests the potential of celecoxib-loaded PEA microspheres to be used as a safe drug delivery system with auto regulatory behavior for treatment of pain associated with OA of the knee

COMMD9 promotes TFDP1/E2F1 transcriptional activity via interaction with TFDP1 in non-small cell lung cancer   

Cellular Signalling

Weihua Zhan, Wenjuan Wang, Tianyu Han, Caifeng Xie, Tingting Zhang, Mingxi Gan, Jian-Bin Wang

COMMD protein family is an evolutionarily conserved gene family implicated in a number of critical processes including inflammation, copper homeostasis, sodium balance, endosomal sorting and cancer. In an effort to profile the expression pattern of COMMD family in several non-small cell lung cancer (NSCLC) cell lines, we found that compared with that in human bronchial epithelial (HBE) cells, the mRNA expression levels of five COMMD genes including COMMD3, COMMD4, COMMD5, COMMD6 and COMMD8 were significantly down-regulated, whereas COMMD9 was up-regulated in NSCLC cell lines. Here we reported that the expression of COMMD9 protein was significantly increased in various NSCLC cell lines and tissue samples. SiRNA-induced knocking down of COMMD9 inhibited proliferation and migration, arrested cell cycle at G1/S transition and induced autophagy in NSCLC cells. Mechanistically, COMMD9 interacted with the TFDP1 through COMM domain, and DNA-binding domain of TFDP1 was required for this interaction. Moreover, decreased expression COMMD9 attenuated TFDP1/E2F1 activation accompanied with enhanced p53 signaling pathway. Taken together, these findings demonstrate that COMMD9 participates in TFDP1/E2F1 activation and plays a critical role in non-small cell lung cancer.

Delineating the Role of βIV-Tubulins in Pancreatic Cancer: βIVb-Tubulin Inhibition Sensitizes Pancreatic Cancer Cells to Vinca Alkaloids   


G. Sharbeen, J. McCarroll, J. Liu, J. Youkhana, L.F. Limbri, A.V. Biankin, A. Johns, M. Kavallaris, D. Goldstein, P.A. Phillips

Pancreatic cancer (PC) is a lethal disease which is characterized by chemoresistance. Components of the cell cytoskeleton are therapeutic targets in cancer. βIV-tubulin is one such component that has two isotypes—βIVa and βIVb. βIVa and βIVb isotypes only differ in two amino acids at their C-terminus. Studies have implicated βIVa-tubulin or βIVb-tubulin expression with chemoresistance in prostate, breast, ovarian and lung cancer. However, no studies have examined the role of βIV-tubulin in PC or attempted to identify isotype specific roles in regulating cancer cell growth and chemosensitivity. We aimed to determine the role of βIVa- or βIVb-tubulin on PC growth and chemosensitivity. PC cells (MiaPaCa-2, HPAF-II, AsPC1) were treated with siRNA (control, βIVa-tubulin or βIVb-tubulin). The ability of PC cells to form colonies in the presence or absence of chemotherapy was measured by clonogenic assays. Inhibition of βIVa-tubulin in PC cells had no effect chemosensitivity. In contrast, inhibition of βIVb-tubulin in PC cells sensitized to vinca alkaloids (Vincristine, Vinorelbine and Vinblastine), which was accompanied by increased apoptosis and enhanced cell cycle arrest. We show for the first time that βIVb-tubulin, but not βIVa-tubulin, plays a role in regulating vinca alkaloid chemosensitivity in PC cells. The results from this study suggest βIVb-tubulin may be a novel therapeutic target and predictor of vinca alkaloid sensitivity for PC and warrants further investigation.

Nanotopography controls cell cycle changes involved with skeletal stem cell self-renewal and multipotency   


Louisa C.Y. Lee, Nikolaj Gadegaard, María C. de Andrés, Lesley-Anne Turner, Karl V. Burgess, Stephen J. Yarwood, Julia Wells, Manuel Salmeron-Sanchez, R.M. Dominic Meek, Richard O.C. Oreffo, Matthew J. Dalby

In culture isolated bone marrow mesenchymal stem cells (more precisely termed skeletal stem cells, SSCs) spontaneously differentiate into fibroblasts, preventing the growth of large numbers of multipotent SSCs for use in regenerative medicine. However, the mechanisms that regulate the expansion of SSCs, while maintaining multipotency and preventing fibroblastic differentiation are poorly understood. Major hurdles to understanding how the maintenance of SSCs is regulated are (a) SSCs isolated from bone marrow are heterogeneous populations with different proliferative characteristics and (b) a lack of tools to investigate SSC number expansion and multipotency. Here, a nanotopographical surface is used as a tool that permits SSC proliferation while maintaining multipotency. It is demonstrated that retention of SSC phenotype in culture requires adjustments to the cell cycle that are linked to changes in the activation of the mitogen activated protein kinases. This demonstrates that biomaterials can offer cross-SSC culture tools and that the biological processes that determine whether SSCs retain multipotency or differentiate into fibroblasts are subtle, in terms of biochemical control, but are profound in terms of determining cell fate.

Systematic Evaluation of Monoclonal Antibodies and Immunoassays for the Detection of Interferon-γ and Interleukin-2 in Old and New World Non-Human Primates   

Journal of Immunological Methods

Ankie Höglind, Irene Areström, Cecilia Ehrnfelt, Khosro Masjedi, Bartek Zuber, Luis Giavedoni, Niklas Ahlborg

Non-human primates (NHP) provide important animal models for studies on immune responses to infections and vaccines. When assessing cellular immunity in NHP, cytokines are almost exclusively analyzed utilizing cross-reactive anti-human antibodies. The functionality of antibodies has to be empirically established for each assay/application as well as NHP species.


A rational approach was employed to identify monoclonal antibodies (mAb) cross-reactive with many NHP species. Panels of new and established mAbs against human Interferon (IFN)-γ and Interleukin (IL)-2 were assessed for reactivity with eukaryotically expressed recombinant IFN-γ and IL-2, respectively, from Old (rhesus, cynomolgus and pigtail macaques, African green monkey, sooty mangabey and baboon) and New World NHP (Ma's night monkey, squirrel monkey and common marmoset). Pan-reactive mAbs, recognizing cytokines from all NHP species, were further analyzed in capture assays and flow cytometry with NHP peripheral blood mononuclear cells (PBMC).


Pan-reactive mAb pairs for IFN-γ well as IL-2 were identified and used in ELISA to measure IFN-γ and IL-2, respectively, in Old and New World NHP PBMC supernatants. The same mAb pairs displayed high functionality in ELISpot and FluoroSpot for the measurement of antigen-specific IFN-γ and IL-2 responses using cynomolgus PBMC. Functionality of pan-reactive mAbs in flow cytometry was also verified with cynomolgus PBMC.


The development of well-defined immunoassays functional with a panel of NHP species facilitates improved analyses of cellular immunity and enables inclusion in multiplex cytokine assays intended for a variety of NHP.

Process Development for Expansion of Human Mesenchymal Stromal Cells in a 50 L Single-use Stirred Tank Bioreactor

Biochemical Engineering Journal

Tristan Lawson, Daniel E. Kehoe, Aletta C. Schnitzler, Peter J. Rapiejko, Kara A. Der, Kathryn Philbrick, Sandhya Punreddy, Susan Rigby, Robert Smith, Qiang Feng, Julie R. Murrell, Martha S. Rook

The application of cell-based therapeutics requires development of refined and scalable culture processes. Stirred tank bioreactors facilitate growth of human mesenchymal stromal cells (hMSC) while meeting these needs. A process for expansion of hMSCs in a 50 L bioreactor was developed. Parameters evaluated include agitation rate, pH and dissolved oxygen (DO) control set-points, and media formulation. The pH and DO levels were determined empirically in 3 L bioreactors prior to implementation at the 50 L scale. The agitation operating range for microcarrier cultures in the 50 L bioreactor was calculated based on theoretical and empirical analyses of solid suspension and shear limitations. Additionally, small-scale experiments demonstrated that hMSC growth was improved in αMEM supplemented with human platelet lysate in comparison to DMEM supplemented with FBS. A 43-fold expansion of harvested hMSCs was achieved in 11 days in a 50 L bioreactor incorporating these process improvements. Cells expanded in the bioreactor exhibited the expected surface marker expression, trilineage differentiation potential, T cell growth inhibition and indoleamine 2, 3-dioxygenase inducibility. The results highlight that identification of optimal pH, DO, agitation rates and culture medium for microcarrier-based bioreactor expansion of adherent cells is paramount to developing a platform to support cell-based therapies.

In-silico and in-vitro anti-cancer potential of a curcumin analogue (1E, 6E)-1, 7-di (1H-indol-3-yl) hepta-1, 6-diene-3, 5-dione   

Biomedicine & Pharmacotherapy

Shamim Akhtar Sufi, Lakshmi Narayana Adigopula, Safiulla Basha Syed, Victor Mukherjee, Mohane S. Coumar, H. Surya Prakash Rao, Rukkumani Rajagopalan

Previously we showed that BDMC, an analogue of curcumin suppresses growth of human breast and laryngeal cancer cell line by causing apoptosis. Here, we demonstrate the enhanced anti-cancer activity of a heterocyclic ring (indole) incorporated curcumin analogue ((1E, 6E)-1, 7-di (1H-indol-3-yl) hepta-1, 6-diene-3, 5-Dione), ICA in short, in comparison to curcumin.


ICA was synthesized by a one pot condensation reaction. Anti-cancer potential of ICA was assessed in three human cancer cell lines of different origin (Lung adenocarcinoma (A549), leukemia (K562) and colon cancer (SW480)) by MTT assay. Mode of cell death was determined by acridine orange-ethidium bromide (Ao-Eb) staining. Putative cellular targets of ICA were investigated by molecular docking studies. Cell cycle analysis following curcumin or ICA treatment in SW480 cell line was carried out by flow cytometry. Expression levels of Cyclin D1 and apoptotic markers, such as Caspase 3, 8 and 9 were studied by western blot analysis in SW480 cell line treated with or without ICA and curcumin.


The yield of ICA synthesis was found to be 69% with a purity of 98%. ICA demonstrated promising anti-cancer activity compared to curcumin alone, as discerned by MTT assay. ICA was non-toxic to the cell line of normal origin. We further observed that ICA is ∼2 fold more potent than curcumin in inhibiting the growth of SW480 cells. Ao-Eb staining revealed that ICA could induce apoptosis in all the cell lines tested. Molecular docking studies suggest that ICA may possibly exhibit its anticancer effect by inhibiting EGFR in A549, Bcr-Abl in K562 and GSK-3β kinase in SW480 cell line. Moreover, ICA showed strong binding avidity for Bcl-2 protein in silico, which could result in induction of apoptosis. Cell cycle analysis revealed that both curcumin and ICA induced concomitant cell cycle arrest at G0/G1 and G2/M phase. Western blot shows that ICA could effectively down regulate the expression of cell cycle protein cyclin D1, while promoting the activation of Caspase 3, 8 and 9 when compared to curcumin in human colon cancer cell line SW480.


The result of this study indicates that ICA could hold promise to be a potential anti-cancer agent. Since ICA has shown encouraging results in terms of its anti-cancer activity compared to curcumin, further research is necessary to fully delineate the underlying molecular mechanism of its anticancer potential.

Inhibition of the MAPK Pathway Alone is Insufficient to Account for all of the Cytotoxic Effects of Naringenin in MCF-7 Breast Cancer Cells

Biochimie Open

Lauren Eanes, Yashomati M. Patel

Estrogen receptor (ER) antagonists such as tamoxifen (Tam) have been used successfully to treat ER+ breast cancers for more than 30 years. Unfortunately, long term use of Tam can result in resistance. Tam resistance is associated with the activation of growth factor signaling pathways that promote cell proliferation and survival. The mitogen activated protein kinase (MAPK), is up-regulated in Tam resistant (Tam-R) cells. Previous studies have reported that the flavanone, naringenin (Nar) can inhibit cell proliferation and induce apoptosis in ER+ breast cancer cells. Furthermore, Nar has been shown to inhibit the MAPK signaling pathways in MCF-7 cells. In this report we investigated whether inhibition of MAPK alone is mediating the effects of Nar on cell proliferation and viability. These studies will determine the mechanism of action of Nar. Tam-R MCF-7 breast cancer cells were treated with Nar or U0126, a MAPK kinase inhibitor. Our studies show that while both U0126 and Nar impaired cell proliferation and viability the combination of U0126 and Nar resulted in greater inhibition of cell viability than either compound alone. It has been previously reported that Nar can bind the ER. Our lab has also shown that Nar localizes ERα to a peri-nuclear region of the cell. Confocal microscopy revealed that in U0126 treated cells ERα displayed an even distribution across the cytoplasm as seen in untreated Tam-R cells. These studies suggest that MAPK is not the only target of Nar.

Glycogen storage disease type Ib neutrophils exhibit impaired cell adhesion and migration   

Biochemical and Biophysical Research Communications

Goo-Young Kim, Young Mok Lee, Joon Hyun Kwon, Hyun Sik Jun, Janice Chou

Glycogen storage disease type Ib (GSD-Ib), characterized by impaired glucose homeostasis, neutropenia, and neutrophil dysfunction, is an inherited autosomal recessive disorder caused by a deficiency in the glucose-6-phosphate transporter (G6PT). Neutrophils play an essential role in the defense against invading pathogens. The recruitment of neutrophils towards the inflammation sites in response to inflammatory stimuli is a tightly regulated process involving rolling, adhesion, and transmigration. In this study, we investigated the role of G6PT in neutrophil adhesion and migration using in vivo and in vitro models. We showed that the GSD-Ib (G6pt−/−) mice manifested severe neutropenia in both blood and bone marrow, and treating G6pt−/− mice with granulocyte colony-stimulating factor (G-CSF) corrected neutropenia. However, upon thioglycolate challenge, neutrophils from both untreated and G-CSF-treated G6pt−/−mice exhibited decreased ability to migrate to the peritoneal cavity. In vitro migration and cell adhesion of G6PT-deficient neutrophils were also significantly impaired. Defects in cell migration were not due to enhanced apoptosis or altered fMLP receptor expression. Remarkably, the expression of the β2 integrins CD11a and CD11b, which are critical for cell adhesion, was greatly decreased in G6PT-deficient neutrophils. This study suggests that deficiencies in G6PT cause impairment in neutrophil adhesion and migration via aberrant expression of β2 integrins, and our finding should facilitate the development of novel therapies for GSD-Ib

The effect of dietary supplementation of algae rich in docosahexaenoic acid on boar fertility   


E.M. Murphy, C. Stanton, C.O.' Brien, C. Murphy, S. Holden, R.P. Murphy, P. Varley, M.P. Boland, S. Fair

The objective of this study was to assess the effects of dietary supplementation of a commercial algal product rich in docosahexaenoic acid (DHA) on boar fertility as assessed in vitro and in vivo. Boars were fed one of three experimental diets for 19 weeks: (i) Control (Ctl) diet (n = 31), (ii) Ctl diet plus 75g All-G-Rich per day (n = 31) or (iii) Ctl diet plus 150g All-G-Rich per day (n = 30). Parameters assessed were (i) raw semen quality; volume, sperm concentration, total motility and morphology (ii) liquid semen quality; progressive motility, viability, hypotonic resistance and acrosomal integrity (iii) frozen-thawed semen quality; motility, thermal stress, viability, membrane fluidity and mitochondrial activity (iv) sperm and seminal plasma (SP) fatty acid composition (FAC) (v) total antioxidant capacity (TAC) of SP and (vi) farrowing rates and litter sizes of sows (n = 1158) inseminated with liquid semen. Boars consuming 75g All-G-Rich had a larger semen volume (P < 0.05) and a higher total sperm number (P < 0.01) than the Ctl treatment, however, there was no effect of treatment on any other semen quality parameter (P > 0.05). There was no effect of dietary treatment on the FAC and TAC of SP or on farrowing rate and litter size (P > 0.05). There was an effect of dietary treatment on the FAC of sperm, represented by an 1.72 and 1.60 fold increase in the DHA content for 75 and 150g treatments, respectively, compared to the Ctl treatment. In conclusion, a significant increase in semen volume and total sperm number in boars supplemented 75g All-G-Rich daily, resulted in an increase in production of 3 to 4 more doses per ejaculate, thus, indicating that the feeding regime described within this study has the potential for increasing the output of boar studs




Title Abstract
Altered expression of CD63 and exosomes in scleroderma dermal fibroblasts

Journal of Dermatological Science, Available online 1 July 2016, Pages

Kayo Nakamura, Masatoshi Jinnin, Miho Harada, Hideo Kudo, Wakana Nakayama, Kuniko Inoue, Aki Ogata, Ikko Kajihara, Satoshi Fukushima, Hironobu Ihn Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto 860-8556, Japan
Exosomes are small vesicles shed from various cells. They contain proteins, lipids, and nucleic acids, and are regarded as a tool of cell-cell communication. To reveal the putative role of exosomes in systemic sclerosis (SSc), and to elucidate the effect of exosomes on wound healing. The expression of common markers for exosomes (CD63, CD9, and CD81) and type I collagen were examined with real-time PCR, immunohistochemical analysis, ELISA, immunoblotting, and flow cytometry. The effect of serum-derived exosomes on wound healing was tested on full-thickness wounds in the mid-dorsal skin of BALB/c mice. The expression levels of CD63 as well as CD9 and CD81 tended to be increased in SSc dermal fibroblasts compared to normal fibroblasts. Increased exosomes in a cultured media of SSc fibroblasts stimulated the expression levels of type I collagen in normal fibroblasts. As the mechanism, collagen-related microRNA levels in SSc fibroblast-derived exosomes were dysregulated, indicating that both the amount and the content of exosomes were altered in SSc. On the other hand, SSc sera showed significantly decreased exosome levels compared to normal sera. The frequencies of vascular involvements, including skin ulcers or pitting scars, were significantly increased in patients with decreased serum exosome levels. The healing of mice wounds was accelerated by treatment with serum-derived exosomes. Vascular abnormalities in SSc may account for the decreased serum exosome levels by the disturbed transfer of exosomes from the skin tissue to the blood stream. Our study suggests the possibility that SSc patients with vascular involvements have decreased serum exosome levels, which causes the delay of wound healing due to down-regulation of collagen, resulting in higher susceptibility to pitting scars and/or ulcers. Exosome research will lead to a detailed understanding of SSc pathogenesis and new therapeutic approaches.
Electrosprayed Montelukast/Poly (Lactic-co-Glycolic Acid) Particle based Coating: a New Therapeutic Approach towards the Prevention of In-stent Restenosis  
Acta Biomaterialia, Available online 7 July 2016, Pages


Maedeh Zamania, Molamma P. Prabhakarana, Jaleh Varshosazb, Priyadarshini S. Mhaisalkarc, Seeram Ramakrishnaa a Department of Mechanical Engineering, National University of Singapore, 2 Engineering Drive 3, Singapore 117576, Singapore b Department of Pharmaceutics, Novel Drug Delivery Systems Research Centre and School of Pharmacy, Isfahan University of Medical Sciences, Isfahan 81746-73461, Iran c School of Materials and Science Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798, Singapore
Drug-eluting stents (DESs), have shown promising results in prevention of in-stent restenosis after percutaneous coronary intervention (PCI). The elevated level of leukotrienes (LTs) detected in injured arteries after PCI, together with the potential role of LTs in inflammatory cascades and structural alterations in arterial wall provides the rationale for development of therapeutic strategies for prevention of in-stent restenosis using LTs receptor antagonists. Montelukast (MK) is a selective cysLT1 receptor antagonist, with anti-inflammatory and anti-proliferative properties, which has been used for treatment of various diseases. Here, we report on the fabrication of MK/PLGA particles by electrospraying, aiming towards the development of particle based coating of DESs. The electrosprayed particles incorporated with 3% and 6% w/w MK exhibited fairly spherical shape with smooth surfaces and narrow size distribution. Sustained release of MK for up to 40 days was obtained for both formulations, with higher initial burst release and drug release rate for the particles with higher drug loading. The LTD4 induced proliferation and migration of human coronary artery smooth muscle cells (HCASMCs) by 35% and 85%, respectively, which was substantially antagonized using MK incorporated particles. Nevertheless, MK antagonism preserved the normal proliferation and migration of human coronary artery endothelial cells (HCAECs). Moreover, MK antagonism inhibited the LTD4 induced phenotypic transition of HCASMCs from contractile to synthetic type. The electrosprayed MK-PLGA particles can be employed as a coating for DESs to inhibit the formation of neointimal hyperplasia responsible for in-stent restenosis, yet preserve the healing rate of the stented vessel.
Design and validation of a consistent and reproducible manufacture process for the production of clinical-grade bone marrow–derived multipotent mesenchymal stromal cells   

Cytotherapy, Available online 14 July 2016, Pages

Margarita Codinach1, Margarita Blanco1, Isabel Ortega1, Mireia Lloret1, Laura Reales1, Maria Isabel Coca1, Sílvia Torrents1, Manel Doral1, Irene Oliver-Vila1, Miriam Requena-Montero1, Joaquim Vives1, Joan Garcia-López1,2 1 Divisió de Teràpies Avançades/XCELIA, Banc de Sang i Teixits, Edifici Dr. Frederic Duran i Jordà, Passeig Taulat, Barcelona, Spain 2 Chair of Transfusion Medicine and Cellular and Tissue Therapies, Universitat Autònoma de Barcelona, Campus UAB, Cerdanyola del Vallès, Bellaterra, Spain
Multipotent mesenchymal stromal cells (MSC) have achieved a notable prominence in the field of regenerative medicine, despite the lack of common standards in the production processes and suitable quality controls compatible with Good Manufacturing Practice (GMP). Herein we describe the design of a bioprocess for bone marrow (BM)–derived MSC isolation and expansion, its validation and production of 48 consecutive batches for clinical use. BM samples were collected from the iliac crest of patients for autologous therapy. Manufacturing procedures included: (i) isolation of nucleated cells (NC) by automated density-gradient centrifugation and plating; (ii) trypsinization and expansion of secondary cultures; and (iii) harvest and formulation of a suspension containing 40 ± 10 × 106 viable cells. Quality controls were defined as: (i) cell count and viability assessment; (ii) immunophenotype; and (iii) sterility tests, Mycoplasma detection, endotoxin test and Gram staining. A 3-week manufacturing bioprocess was first designed and then validated in 3 consecutive mock productions, prior to producing 48 batches of BM-MSC for clinical use. Validation included the assessment of MSC identity and genetic stability. Regarding production, 139.0 ± 17.8 mL of BM containing 2.53 ± 0.92 × 109 viable NC were used as starting material, yielding 38.8 ± 5.3 × 106 viable cells in the final product. Surface antigen expression was consistent with the expected phenotype for MSC, displaying high levels of CD73, CD90 and CD105, lack of expression of CD31 and CD45 and low levels of HLA-DR. Tests for sterility, Mycoplasma, Gram staining and endotoxin had negative results in all cases. Herein we demonstrated the establishment of a feasible, consistent and reproducible bioprocess for the production of safe BM-derived MSC for clinical use.
Wenshen Xiaozheng Tang induces apoptosis and inhibits migration of ectopic endometriotic stromal cells
Journal of Ethnopharmacology, Available online 9 July 2016, Pages

Zhenzhen Zhang a,b, Xiaolan Cheng a,b, Tao Gui a,b, Jia Tao a,b, Meihua Huang a,b, Li Zhu a,b, Mei Luo a,b, Peng Cao a,b, Guiping Wan a,b a Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, 210028, Jiangsu, P.R. China b Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing, 210028, Jiangsu, P.R. China
Wenshen Xiaozheng Tang (WXT), a traditional Chinese medicine prescription, exerted a good therapeutic effect on endometriosis. However, the underlying mechanism is unclear. In the present study, we sought to evaluate the effect of WXT on the proliferation and migration of ectopic endometriotic stromal cells and explore the potential molecular mechanism. Primary stromal cells derived from ectopic endometriotic lesions of patients with endometriosis were isolated and cultured. The inhibition effect of WXT on cell proliferation was determined by MTT. Apoptosis of ectopic endometriotic cells treated with WXT was analyzed with Annexin V-FITC/7-AAD staining. The activation of caspases was detected by western blot analysis. The influence of WXT on migration of ectopic endometriotic cells was measured by scratch wound healing assay and Transwell assay. The DNA binding activity of NF-κB and the expression of nuclear p65 protein were determined by electrophoretic mobility shift assay and western blot analysis, respectively. The impact of WXT on the expression of NF-κB regulated gene products involved in apoptosis and migration was determined by western blot analysis. WXT inhibited the proliferation of ectopic endometriotic cells in a time- and dose-dependent manner. In addition, WXT treatment resulted in significant induction of apoptosis through the activation of caspases and inhibition of migration in ectopic endometriotic cells. WXT notably suppressed constitutive NF-κB-DNA-binding activity as well as TNF-α induced nuclear translocation of NF-κB p65 subunit in ectopic endometriotic cells. Moreover, WXT diminished the expression of NF-κB regulated gene products involved in apoptosis and migration, including c-IAP1, c-IAP2, XIAP, survivin, Mcl-1, COX-2 and MMP-9. Our results indicate that WXT induces apoptosis and inhibits migration of ectopic endometriotic stromal cells.
A combination of an iron chelator with an antioxidant effectively diminishes the dendritic loss, tau-hyperphosphorylation, amyloids-β accumulation and brain mitochondrial dynamic disruption in rats with chronic iron-overload  

Neuroscience, Available online 9 July 2016, Pages

Jirapas Sripetchwandee a,b, Suwakon Wongjaikam a,b, Warunsorn Krintratun a,b, Nipon Chattipakorn a,b, Siriporn C. Chattipakorn a,c a Neurophysiology Unit, Cardiac Electrophysiology Research and Training Center, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand b Department of Physiology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand c Department of Oral Biology and Diagnostic Sciences, Faculty of Dentistry, Chiang Mai University, Chiang Mai 50200, Thailand
Iron-overload can cause cognitive impairment due to blood–brain barrier (BBB) breakdown and brain mitochondrial dysfunction. Although deferiprone (DFP) has been shown to exert neuroprotection, the head-to-head comparison among iron chelators used clinically on brain iron-overload has not been investigated. Moreover, since antioxidant has been shown to be beneficial in iron-overload condition, its combined effect with iron chelator has not been tested. Therefore, the hypothesis is that all chelators provide neuroprotection under iron-overload condition, and that a combination of an iron chelator with an antioxidant has greater efficacy than monotherapy. Male Wistar rats (n = 42) were assigned to receive a normal diet (ND) or a high-iron diet (HFe) for 4 months. At the 2nd month, HFe-fed rats were treated with a vehicle, deferoxamine (DFO), DFP, deferasirox (DFX), n-acetyl cysteine (NAC) or a combination of DFP with NAC, while ND-fed rats received vehicle. At the end of the experiment, rats were decapitated and brains were removed to determine brain iron level and deposition, brain mitochondrial function, BBB protein expression, brain mitochondrial dynamic, brain apoptosis, tau-hyperphosphorylation, amyloid-β (Aβ) accumulation and dendritic spine density. The results showed that iron-overload induced BBB breakdown, brain iron accumulation, brain mitochondrial dysfunction, impaired brain mitochondrial dynamics, tau-hyperphosphorylation, Aβ accumulation and dendritic spine reduction. All treatments, except DFX, attenuated these impairments. Moreover, combined therapy provided a greater efficacy than monotherapy. These findings suggested that iron-overload induced brain iron toxicity and a combination of an iron chelator with an antioxidant provided a greatest efficacy for neuroprotection than monotherapy.
A Screen of FDA-Approved Drugs for Inhibitors of Zika Virus Infection   

Cell Host & Microbe, Available online 28 July 2016, Pages

Nicholas J. Barrows1,2, Rafael K. Campos1,2, Steven T. Powell1, K. Reddisiva Prasanth1, Geraldine Schott-Lerner1, Ruben Soto-Acosta1, Gaddiel Galarza-Muñoz1, Erica L. McGrath3, Rheanna Urrabaz-Garza4, Junling Gao3, Ping Wu3, Ramkumar Menon4, George Saade4, Ildefonso Fernandez-Salas5, Shannan L. Rossi6, Nikos Vasilakis6, Andrew Routh1,7, Shelton S. Bradrick1, Mariano A. Garcia-Blanco1,8 1 Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, 301 University Blvd, Galveston, TX 77555, USA 2 Department of Molecular Genetics and Microbiology, Duke University, Durham, NC 27710, USA 3 Department of Neuroscience and Cell Biology, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA 4 Department of Obstetrics and Gynecology, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA 5 Centro Regional de Investigación en Salud Publica INSP, 19 Poniente Esquina 4a Norte s/n, Colonia Centro, Tapachula, Chiapas 30700, C.P., Mexico 6 Department of Pathology, Center for Biodefense and Emerging Infectious Diseases, Center for Tropical Diseases, and Institute for Human Infections and Immunity, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA 7 Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, 301 University Blvd, Galveston, TX 77555, USA 8 Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169857, Republic of Singapore
Currently there are no approved vaccines or specific therapies to prevent or treat Zika virus (ZIKV) infection. We interrogated a library of FDA-approved drugs for their ability to block infection of human HuH-7 cells by a newly isolated ZIKV strain (ZIKV MEX_I_7). More than 20 out of 774 tested compounds decreased ZIKV infection in our in vitro screening assay. Selected compounds were further validated for inhibition of ZIKV infection in human cervical, placental, and neural stem cell lines, as well as primary human amnion cells. Established anti-flaviviral drugs (e.g., bortezomib and mycophenolic acid) and others that had no previously known antiviral activity (e.g., daptomycin) were identified as inhibitors of ZIKV infection. Several drugs reduced ZIKV infection across multiple cell types. This study identifies drugs that could be tested in clinical studies of ZIKV infection and provides a resource of small molecules to study ZIKV pathogenesis.
Development and validation of a cell-based assay system to assess human immunodeficiency virus type 1 integrase multimerization

Original Research Article
Journal of Virological Methods, Available online 26 July 2016, Pages

Tomofumi Nakamuraa, Joseph R. Campbella, Amber R. Moorea, Sachiko Otsua, Haruo Aikawab, Hirokazu Tamamurab, Hiroaki Mitsuyaa,c,d a Departments of Infectious Diseases and Hematology, Kumamoto University Graduate School of Medical Sciences, Kumamoto 860-8556, Japan b Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Chiyoda-ku, Tokyo, 101-0062, Japan c Experimental Retrovirology Section, National Center for Global Health and Medicine Research Institute, Shinjuku, Tokyo 162-8655, Japan d HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
Multimerization of HIV-1 integrase (IN) subunits is required for the concerted integration of HIV-1 proviral DNA into the host genome. Thus, the disruption of IN multimerization represents a new avenue for intervening HIV-1 infection. Here, we generated a cell-based assay system to assess IN multimerization using a newly constructed bimolecular fluorescence complementation (BiFC-IN) system. BiFC-IN proteins were efficient in emitting fluorescence, and amino acid (AA) substitutions associated with IN multimerization attenuated fluorescence, suggesting that the BiFC-IN system may be useful for evaluating the profile of IN multimerization. A recently reported non-catalytic site IN inhibitor (NCINI), which allosterically induces IN over-multimerization/aggregation, significantly increased fluorescence in the BiFC-IN system. An IN's substitution, A128T, associated with viral resistance to NCINIs, decreased the NCINI-induced increase of fluorescence, suggesting that A128T reduces the potential for IN over-multimerization. Moreover, E11K and F181T substitutions known to inhibit IN tetramerization also reduced the NCINI-induced fluorescence increase, suggesting that NCINI-induced IN over-multimerization was more likely to occur from tetramer subunits than from dimer subunits. The present study demonstrates that our cell-based BiFC-IN system may be useful in elucidating the profile of IN multimerization, and also help evaluate and identify novel compounds that disrupt IN multimerization in living cells.
Effects of different ligands on epidermal growth factor receptor (EGFR) nuclear translocation   

Biochemical and Biophysical Research Communications, Available online 25 July 2016, Pages

Jerusa A.Q.A. Faria a, Carolina de Andrade a, Alfredo M. Goes a, Michele A. Rodrigues a,b, Dawidson A. Gomes a a Department of Biochemistry and Immunology, Universidade Federal de Minas Gerais, Av. Antonio Carlos, 6627, Belo Horizonte, MG, 31270-901, Brazil b Department of General Pathology, Universidade Federal de Minas Gerais, Av. Antonio Carlos, 6627, Belo Horizonte, MG, 31270-901, Brazil
The epidermal growth factor receptor (EGFR) is activated through binding to specific ligands and generates signals for proliferation, differentiation, migration, and cell survival. Recent data show the role of nuclear EGFR in tumors. Although many EGFR ligands are upregulated in cancers, little is known about their effects on EGFR nuclear translocation. We have compared the effects of six EGFR ligands (EGF, HB-EGF, TGF-α, β-Cellulin, amphiregulin, and epiregulin) on nuclear translocation of EGFR, receptor phosphorylation, migration, and proliferation. Cell fractionation and confocal immunofluorescence detected EGFR in the nucleus after EGF, HB-EGF, TGF-α and β-Cellulin stimulation in a dose-dependent manner. In contrast, amphiregulin and epiregulin did not generate nuclear translocation of EGFR. EGF, HB-EGF, TGF-α and β-Cellulin showed correlations between a higher rate of wound closure and increased phosphorylation of residues in the carboxy-terminus of EGFR, compared to amphiregulin and epiregulin. The data indicate that EGFR is translocated to the nucleus after stimulation with EGF, HB-EGF, TGF-α and β-Cellulin, and that these ligands are related to increased phosphorylation of EGFR tyrosine residues, inducing migration of SkHep-1 cells.