Scientific Program

Sessions:

Cell Science & Molecular Biology

Abstract

In collaboration with the colleagues in Bone Marrow Transplantation Processing Laboratories of Medical College of Wisconsin and Froedtert Memorial Lutheran Hospital (Milwaukee, WI, USA), we have developed and modified the strategy used for differentiation and maturation of fast monocyte-derived dendritic cells (fast-DCs) within only 48-72 hours of in vitro culture to be effective as mature standard-DCs (generated in 7-10 days of in vitro culture) in priming and propagation of antigen-specific T-cell responses. The use of fast-DCs not only reduces labor and supply cost, as well as workload and time, but also increases the DCs yield from monocytes, which may facilitate DCs-based vaccination for infectious and neoplastic diseases. Novel approaches for generation, pulsing with different antigen sources, and transduction with adenoviral vector of fast-DCs as well as using of fast-DCs for priming and propagation of antigen-specific cytotoxic T-cell (CTL) effectors and expansion of human γδ and CD56+ T-cells will be discussed here. Also, the feasibility of a new approach by which antigen-specific T-cell lines are generated by sequential primings with antigen-pulsed fast-DCs followed by priming and expansion using antigen-pulsed Epstein Barr virus-transformed B lymphoblastoid cell lines (BLCL) to overcome the limitation of CTL effectors expansion imposed by the lack of sufficient DCs will be discussed here. Hopefully, the protocols described here will be of a practical value for generation of fast-DCs based cellular vaccination as adjunct and, more importantly, as prophylaxis against common infectious and neoplastic diseases in high-risk immunocompromized patients, particularly recipients of hematopoietic stem cells transplant

Biography

Prof. Dr. Gamal Ramadan has completed his PhD in Molecular Immunology from, Hannover University (Germany) and postdoctoral studies from Medical College of Wisconsin (USA). He is the head of Physiology Branch, Zoology Department, Faculty of Science, Ain Shams University. Also, he got the Research Paper Award in Basic Science (Wisconsin Cancer Centre, 2004), the State Incentive Award in Immunology (Egyptian Government, 2006), and the Highly Cited Researcher Award (Thomson Reuters, 2016). He has published more than 25 papers in reputed journals and has been serving as an editorial board member and reviewers of repute.

Speaker
Gamal Ramadan Ain Shams University, Cairo, Egypt

Abstract

During immunogenesis B-lymphocytes under antigen pressure transform into antibody-producing (plasma) cells. It is assumed that the strongest immune reactions are provoked by parasites spending all or most of their life cycle in hosts' tissues. This means importance of studies of immune competent spleen cells' role in the hosts' response to Trichinella infection. Material and Methods. Experiments used 25 outbred white rats 5-6 months old weighting 230-250 g, 20 of which were infected with 10 l/g body mass of Trichinella spiralis larvae, and 5 were control. Rats were decapitated at 4, 6, 26, 38 days p.i. Spleen stamps were stained using Pappenheim method and plasma line cells counted in 50 microscope view fields. Results. At 4th day p.i. a statistically significant increase in plasma cells numbers occurred. Their numbers in spleen stains raised 2.3 times relative to control (37.0±0.8, in control – 16.0±0.7) and this occurred through plasmablasts and proplasmacytes. Probably Trichinella antigens stimulate lymphocytes and contribute to their proliferation and differentiation. This period corresponds with intestinal stage of the parasite's development. Later, at 7th day, plasma reaction suppression occurred, so proliferation level and numbers of plasma cells lowered (14,0±0,45). This is due to Trichinella larvae having highest biological activity during blood migration when they grow, molt , excreting antigens into blood. At 30th day p.i. (muscle stage of parasite's development) a repeat two times increase in plasma cells numbers occurred. Continued observations (38 d.p.i.) didn't show significant changes in plasma cells numbers. The work was supported by RSF, grant 14-16-00026

Biography

S.O. Movsesyan has completed his Candidate degree at 1962 and Doctor degree at 1971 from Skrjabin Helminthology Institute, Moscow, USSR. He is former director of Institute of Zoology of Armenia and Institute of Parasitology RAS, renowned scientific organizations. He has published more than 450 papers in reputed journals and has been serving as an editorial board member of repute.

Speaker
S. Movsesyan A.N. Severtsov INstitute of Ecology and Evolution, Russia

Abstract

Despite significant advances in medical sciences, cancer is still an uncontrollable disease in humans and is a major cause of death around the world. Therefore, identification of novel potent, selective, less toxic and apoptotic inducer anticancer agents is so important in the world of science. The roles of Reactive Oxygen Species (ROS) and Nitric Oxide (NO) in Programmed cell death (apoptosis) have been reported. The purpose of the current study was to evaluate the production of Reactive Oxygen Species (ROS) and Nitric Oxide (NO) by pyrimidin derivatives in MDA-MB231 and PC3 cell lines. In order to investigate nitric oxide level in MDA-MB231 and PC3 treated cell lines, we measured nitrite (NO) which is one of two primary and nonvolatile breakdown products of NO by Griess reaction method. This assay relies on a diazotization reaction. The amount of ROS in treated cells were conducted using 2’,7’-dichlorfluorescein-diacetate (DCFH-DA) compound. DCFH-DA (nonfluorescent compound) is converted to DCF (fluorescent compound) through the action of peroxid rated by the presence of peroxidase and consequently could be detected. The results of this study indicated that some pyrimidin derivatives with benzochrome groups could induce the production of ROS and NO in MDA-MB231 and PC3 cell lines as comparable or higher than reference drug etoposide and as a result, these compounds could effective in apoptosis induction in treated cells. Benzochromenopyrimidin derivatives were effective on two cell lines and enhanced the level of ROS and NO in treated cells. There has been proved that the enhancement of ROS and NO level could effective in induction of apoptosis. However, the details and mechanisms by which ROS and NO interplay and induce apoptosis are not well understood. MDA-MB231 is a triple-negative breast cancer cell line (TNBC).TNBC cell lines do not respond to endocrine therapy or other available targeted agents. On the other hand, PC3 is a prostate cancer cell line without androgen receptor(AR) and prostate-specific antigen (PSA) and consequently these cells do not respond to hormonal therapy. So maybe benzochromenopyrimidin derivatives could resolve these treatment challenges and may be used as a drug for cancer therapy in future. Key words: Breast cancer, Prostate cancer, ROS, NO, Pyrimidin

Biography

Maliheh Safavi has completed her PhD from Tehran University, and postdoctoral studies from Tehran University of Medical Sciences, Iran. He is an assistant professor in department of Biotechnology, Iranian Research Organization for Science and Technology, Tehran. He has published more than 75 papers in reputed journals and has been serving as reviewer member of repute. She is involved in several international projects as principle investigator.

Speaker
Maliheh Safavi University of Tehran,Iran

Abstract

Caspases, as the key effector molecules in apoptosis, are potential targets for pharmacological modulation of cell death. Increased caspase activity is often observed in the damaged cells in a number of diseases, including myocardial infarction, stroke, sepsis, Alzheimer’s, Parkinson’s, and Huntington’s diseases. Under such conditions, inhibition of caspase activity is predicted to be therapeutically beneficial. In a search for new compounds with such a potential, we tested the effect of ethanolic extract of Boswellia gum resin on caspase-9 activity, the initiator enzyme in the activation of the intrinsic apoptosis pathway. Recombinant caspase-9 was expressed in Escherichia coli and purified with Ni-NTA affinity chromatography. Caspase activity was measured in the presence of various concentrations of gum resin extract of Boswellia species. The extract exhibited potent inhibitory effect on caspase-9 activity. To obtain some insight into the size of the effective ingredient(s), filtration through membranes with defined sizes was performed showing that the inhibitor(s) is smaller than 5 kDa. To determine whether or not the inhibitor is stable, and the sample was heated at 70 °C for 60 minutes without any effect. We further tested dominant ingredients present in Boswellia, namely α- and β-boswellic acids, and found that these are not the inhibitors of caspase-9. Further investigations are under way to identify ingredient caspase-9 inhibitors present in Boswellia.

Biography

Raheleh Shakeri has completed her PhD from University of Tehran, Iran. She is an assistant professor of biochemistry at University of Kurdistan since 2016 till now. Her research during MSc and PhD has focused on intrinsic apoptosis pathway.

Speaker
Raheleh Shakeri University of Kurdistan,, Sanandaj, Iran

Abstract

Cell Sciences and Omics : Perspectives and prospect in Human Health Care

Biography

Mishra received his B. S (Honors) and M.S degrees from Patna University ( then nicknamed as Oxford of the East) in India and Ph. D Degree from McMaster University. He received his post –doctoral training with the late Nobel Laureate Professor E. L. Tatum at the Rockefeller University. He was a Fellow for Medical Research of the Jane Coffin Child Fund of the Yale University at the Rockefeller University for two years and then Research Associate with Professor Tatum where he initiated his work in what is now called as Proteomics and Matabolimics. There he also devised the first gene transfer in a eukaryote, Neurospora crassa.

Speaker
Nawin Mishra The University of South Carolina, Columbia,USA

Abstract

Cardiomyopathy attends from changes in calcium signaling to inflammation to cardiomyocyte toxicity. It is aberrant change that causes mitochondrial pathogenesis, that inhibits the expression of nuclear and mitochondrial encoded genes that is involved in mitochondrial biogenesis. Acetyl Salicylic acid/Aspirin is used in the treatment of atherosclerotic diseases. It is ethyl ester that inhibits cyclooxygenase action to prevent inflammation. We hypothesized that aspirin treatment has ability to induce Sirtuin 1, 4 (Sirt1 and Sirt4) and Sirt1 transcriptional targets, includes peroxisome proliferator-activated receptor-gamma co-activator-1α (PGC1α) and mitochondrial transcription factor A (Tfam) genes in cultured HL1 cardiomyocytes. Studies on cultured HL1 cardiomyocytes showed aspirin treatment induced Sirt1 and Sirt4 genes via; H2O2 generation. However, a decrease in AhR gene indicates that aspirin is hydrolyzed metabolically to Salicylic acid but does not enzymatically forms Dihydroxybenzoates. Increase in Sirt1 gene activated PGC1α and Tfam gene. Aspirin also induced antioxidant enzymes, glutathione peroxidase (GPX) and catalase (CAT) gene. It can be concluded that it is potent anti-atherosclerotic drug and increase in Sirt1 gene further suggest that it can induce anti-inflammatory action. Moreover, absence of AhR gene in heart can induce additional benefits by increasing anti-inflammatory action. It can interfere with Nfkb signaling pathway that can prevent foam cell formation. Increase in fos gene in previous studies may also provide substantial benefits to heart as it can increase tyrosine phosphorylation and may have inotropic effect. However, specific overexpression of PGC1α and Tfam gene in HL1 cardiomyocyte might increase mitochondrial biogenesis and prevent the development of heart failure.

Biography

Speaker
Pratibha Kamble Ohio State University, USA

Abstract

3D culture systems are becoming crucial scientific tools in cancer cell research. They are a bridge between traditional culture on plastic and in vivo experiments. Here, we report on a detailed comparison of cytotoxicity effects of novel derivatives of pyridine on spheroid model of one human breast cancer cell line (MDA-MD-231) with the traditional monolayer culture. To investigate the efficacy of compounds on cell’s mortality and metabolic activity, MTT test were utilized on 2D culture and 3D culture. For 2D culture cells were cultured at 96 well plate. After one day each well was added with 200 μl of freshly medium containing compounds with concentration range. After harvesting the cells, the absorption was read at 492 nm. 3D culture cell system was performed as spheroids in the appropriate media by modification of the hanging drop method and liquid overlay cultivation technique. Results of the MTT assay in 2D culture revealed (E)-2-(4-methoxystyryl)-N-tert-butyl-6-methylH-imidazo[1,2-a]pyridin-3-amine showed potent activity against MDA-MB-231cells in comparison to standard drug etoposide . In 3D culture the IC50 values of this compound was 5 times higher than the IC50 values of them in the 2D culture. The IC50 of etoposide was 2 times higher than the IC50 in 2D culture. The results revealed that the cytotoxicity of compounds in 3D culture was reduced in comparison to 2D culture. In 2D culture system, wide surface of cells are exposed to the compounds, but cells located on the outer layer of spheroids decreases the penetration of drugs in to center of spheroids. Keywords: pyridine derivative, (3D) cell culture, cytotoxicity

Biography

Prof. Ardestani is a member of scientific and the research team at Institute of Biochemistry and Biophysics, University of Tehran from 1996. She is teaching Biochemistry of signal transduction and regulation, Immunochemistry and Immunobiology to graduate students (PhD and MS). She was principal investigator of several projects that some of them were National or international projects. She supervised several graduate students thesis. Her interest focused on mechanism of action of medicinal herbs or new synthetic compounds on cancer cells; in the last decade we were able to publish more than 50 papers in scientific journals.

Speaker
Sussan K. Ardestani University of Tehran, Iran

Abstract

Background: Pb was found to produces a wide range of toxic effects including neurotoxicity to the young offspring. The current study was to investigate the beneficial role of garlic in the protection from the maternal and fetal cerebellar damage that produced by Pb during pregnancy. Materials and Methods: Positively pregnant female rats were divided into five groups; one control group, two Pb-treated groups (exposed to 160 and 320 mg/kg b.wt. of Pb, respectively) and two groups treated with both Pb and garlic (exposed to Pb as previous groups together with 250 mg/ kg b.wt. /day of garlic extract). Treatments started from day 1 till day 20 of pregnancy, where the mother rats of different experimental groups were sacrificed to obtain the fetuses. Results: The results showed that administration of Pb to pregnant rats resulted in a dose-dependent toxicity for both mothers and fetuses in the form of decrease of maternal weight gain, placental and fetal weights, brain weight and diminished fetal growth parameters, which were prominent in rat’s group treated with larger dose of Pb. In Pb-treated rats, Pb level in blood and cerebellum was high when compared to the control. Co-treatment of garlic extract along with Pb resulted in a significant decrease in Pb levels as compared with those treated with Pb alone with improvement of the histopathological changes. Conclusions: This study was useful in evaluating hazardous of Pb neurotoxicity due to exposure during pregnancy. Co-administration of garlic has beneficial effects in amelioration of Pb-neurotoxicity of mother and fetuses.

Biography

Dr. Hesham N. Mustafa has received his MD in Basic Medical Sciences (Anatomy) from Ain Shams University, Cairo, Egypt at 2009. Currently, he is working as an Associate Professor in Basic Medical Sciences (Anatomy) department, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia. He has published more than 15 papers in reputed journals and been serving as an editorial board member and a reviewer at many reputed Journals. He received Certificate for Highly Cited Paper from Elsevier in December, 2016 And Award of Excellence of Scientific Publication for the staff members, from Deanship of Scientific Research, King Abdulaziz University.

Speaker
Dr. Hesham N. Mustafa King Abdulaziz University, Jeddah, Saudi Arabia

Abstract

Several genetic, molecular, epigenetic findings have been suggested to explain the role of dyanamic adaptations and alterations within the the cells and their outside nices. These network of genetic, molecular and epigenetic events have been central to various intricacies of life and death, cellular responses external clues. However, several questions are still remaining unsettled with reference to cellular signaling adaptations with reference to new forms of external clues in the form of social networking, which is completely different from the well studied environmental factors such as pathogens, chemical signals, physical signals and stress signals. Therefore, understanding the influences of social networking such as faceboo, internet, instagrams and many more based generated signals to shape up the adaptation of huma intra- and inter cellular signaling adapations may reveal new facets of crosstalk at the level og genetic, epigenetic and social networking driven adaptations. Currently, it is not knwon and possibly clear evidences are lacking whether signals/clues emanating from scoial networking driving changes at the interface of molecular signaling, genetic and epigeentic level. Of couse, if there are any changes, then these changes in intra- and inter cellular signaling network could be linked to the pathophysiological state either pro-disease or anti-disease condition. It appears interesting to extend hypothesis to cancer pathophysiology and its treatment moudlations due to the possible crosstalk between social networking and intra- and inter cellular networking in a cancer patient. In future perspectives, state of the art tools and techniques to decipher alterations in the cellular signaling landscape at molecular, genetic and epigenetic landscape may provide answers to above proposed concept and could pave the way of better understanding of social adaptions of users of social networking tools, and if any impact on the human disease conditions for an example cancer pathophysiology. The author suggests that system biology, transcriptomics, proteomics and epigenetic tools could be performed on animal model or human model to observe changes in the cellular signaling wiring due to the social networking emanated clues/signals. The impact of this hypothesis and experimental apporcahes may help to achieve milestone towards personalized medicine. Since, in the current era of drug development, contribution of external signals and factors in the form of social networking should be highlighted and emphasized for any drug regimen to patients. In this hypothesis, we summarize the importance of social networking to drive intra- and inter-cellular signaling adaptations in the perspective of fundamental of evolution of human cellular organization, personalized medicine and biomarkers of if any disorders associated with the negative imptact of social networking. Key Words: Epigenetic events, Life, death, Cancer, Gene, Epigenome, Environment, Social networking, Cellular signaling

Biography

"Dr. Nilesh Kumar Sharma is a Associate Professor, Department of ""Biotechnology in D.Y. Patil Biotechnology & Bioinformatics Institute, India. His area of specialization is DNA repair and drug resistance in cancer and his experimental methods are Human carcinoma, normal cell line and carcinoma patient tissue."

Speaker
Dr. Nilesh Kumar Sharma Dr. D. Y Patil Vidyapeeth, Pune, India

Abstract

Nuclear receptors comprise a large family, with the human and mouse families containing 46 members. Studies of five steroid hormone receptors established the idea of nuclear receptors as so-called “ligand-activated transcription factors”. However, most other nuclear receptors are non-steroidal, constitutively activated and, and possess no known biological ligands. Thus, understanding the diverse functions and regulations of nuclear receptors requires concepts far beyond ligand activation. Out of these 46 members, 41 conserve a phosphorylation motif within their DNA binding domains in both human and mouse receptors, which has long been forgotten in the history of nuclear receptors. Now phosphorylation at this conserved motif has been shown with four nuclear receptors (ER, CAR, FXR and RXR) in mouse tissues and cells, for which regulatory functions and mechanisms have begun to appear. Phosphorylation uniquely regulates each of these four nuclear receptors. For example, CAR inactivates its constitutive activity by phosphorylation, enabling dephosphorylation to reactivate CAR in response to drugs such as phenobarbital. Phosphorylated ER acquires its anti-inflammatory function in brain microglia. Whereas these observations could be expected, the more intriguing question is why phosphorylation is conserved throughout nuclear receptors. Our hypothesis is that this conserved phosphorylation can be utilized in a communication process through which one nuclear receptor transduces its signal to other nuclear receptors when this receptor requires them to elicits its function. To this end, we are investigating the phenobarbital-induced mechanism that activates estrogen sulfotransferase and cytochrome P450 genes, in which multiple nuclear receptors appear to communicate through their corresponding phosphorylation.

Biography

Dr. Negishi has been Principle Investigator heading the Pharmacogenetics section since 1983 at National Institute of Environmental Health Sciences, National Institutes of Health and is a member of the Senior Biomedical Research Service. He received his Doctor in Science in Biochemistry in 1973 from Institute for Protein Research at Osaka University in Japan and carried out postdoctoral training in the Department of Cell Biology at New York University and National Institute of Child Health and Human Development, NIH His major research interests lie in the regulation mechanisms of drug-induced gene expression and nuclear receptor signaling.

Speaker
Dr.Negishi, Masahiko Osaka University , Japan

Sessions:

Cell Science & Molecular Biology

Abstract

Purinergic P2Y receptors are expressed in the apical and basolateral membranes of virtually all polarized epithelia. Extracellular nucleotides such as UTP and ATP can be released from epithelial cells into the extracellular space and act in an autocrine or paracrine fashion. Extracellular nucleotides act on membrane-bound receptors, termed P2 receptors, which can be classified into two classes: ionotropic P2X receptors and metabotropic P2Y receptors. Activation of P2Y receptors (e.g., P2Y1, P2Y2, P2Y4, and P2Y6) activates the phospholipase C - IP3 - Ca2 signaling pathway. We identify that P2Y6 receptors in bronchial epithelia are doubly coupled to both Ca2 -dependent and cAMP-dependent signaling pathways, both of which are involved in the regulation of epithelial transport by activating different Cl- and K channels located at the apical and basolateral membranes. In addition to the regulation of epithelial transport, P2Y receptors have been implicated in various inflammatory disorders, such as asthma. We demonstrate that when human bronchial epithelia are chemically damaged by poly-L-arginine as a surrogate of cationic proteins, nucleotides are released into the extracellular medium in a polarized fashion. The extracellular nucleotides then activate cell-surface P2Y receptors to release proinflammatory cytokines (IL-6 and IL-8) via a Ca2 -dependent process. Interestingly, P2Y receptor-mediated Ca2 signaling and cytokine secretion can be inhibited by G protein�coupled estrogen receptor�mediated activation of a cAMP-dependent pathway. In summary, purinergic signaling mechanisms have been shown to play key roles in the regulation of many aspects of epithelial function, such as the regulation of ion transport and immune response.

Biography

Dr. Wing-hung Ko obtained his PhD from Department of Physiology, The Chinese University of Hong Kong in 1992. Following postdoctoral training at Institute of Physiology, University of Glasgow, he joined his alma mater as lecturer in 1993. He is currently an Associate Professor of School of Biomedical Sciences. Dr. Ko’s recent research examines P2Y receptor-regulated Ca2 signaling and ion transport in airway epithelia, with a focus on the role of P2Y receptors in airway inflammation. He has over 70 articles published in reputable international journals, such as Journal of Biological Chemistry and American Journal of Respiratory Cell and Molecular Biology.

Speaker
Wing-hung KO The Chinese University of Hong Kong, Hong Kong

Abstract

BackgroundDespite the prevalence and detrimental effect of urinary and fecal incontinence on many postmenopausal women, there seems to be no consensus at present on the exact underlying mechanism. The theory that low circulating estrogen is causally related to these disorders is not based on any supportive evidence. It is not also clear whether normative ageing is primarily or secondarily responsible. Introduction Laboratory animals which are old and those with surgically-induced ovarian failure provide the unique opportunity to investigate the role of these 2 specific biological conditions (menopause and ageing) on the pelvic floor structures circumventing several extrinsic variables inherent in clinical studies. The anatomy of these structures in rats is sufficiently similar to that of humans to serve as a model for experimental studies. Fisher 344 rats are particularly suitable for ageing research because the mean survival time of animals in this colony is prolonged (30 months) compared to other rat models. ObjectivesThe experimental evidence of ageing of the female pelvic floor will be described. MethodsThe most common surrogate biomarkers studied to assess the effects of ageing and/or estrogen deficiency on the pelvic floor were morphological: the amount and relative proportion of submucosal collagen fibers types I and III in the urethra and anal canal and of isomyosin fibers types I and II in the striated pelvic floor muscles using Western Blot analysis as well as the number of submucosal vascular plexuses in the urethra and anal canal using light microscopy. Measurement of cytoplasmic expression of p27kip1 in the striated pelvic floor muscles by Western blot analysis is also recently considered a specific biochemical marker of pelvic floor ageing because this protein normally regulates muscle cell differentiation and apoptosis. The effects of estrogen and/or ghrelin (an anti-ageing preparation widely used in ageing research) administration on these surrogate biomarkers in old versus young adult ovariectomized Fisher 344 rats were compared.ResultsAll biomarkers of pelvic floor ageing were significantly increased in old compared to young-adult female rats. Ovariectomy significantly increased these changes further in old versus young-adult rats with either smaller or larger differential effect on some pelvic floor structures than ageing compared to young-adult animals. The effects of ovariectomy in old rats were totally reversed by combined estrogen/ghrelin replacement rather than by estrogen alone suggesting an independent ageing effect.ConclusionsOvariectomy and subsequent estrogen deprivation exacerbates the inherent ageing changes in the pelvic floor of old rats indicating a synergistic, deleterious and independent interaction between hypoestrogenism and normative ageing. Combined estrogen and ghrelin administration restored these changes in old but not young adult ovariectomized rats through a direct receptor-mediated effect. Our studies thus showed anatomical, histological and ultrastructural evidence of derangement of structure of the female pelvic floor with the cumulative effect of menopause and ageing. Studying the individual contribution of biological versus reproductive senescence on the pelvic floor in animal models may assist in identifying therapeutic strategies to delay or prevent this inevitable process in old post-menopausal women.

Biography

Speaker
Diaa E.E. Rizk Arabian Gulf University, Manama, Bahrain

Abstract

An anectodotal observation reports that in spite of the high degree of vascularization and location throughout the body, skeletal muscle is rarely a site of metastatic tumor formation. The mechanism underlying the resistance of skeletal muscles to metastasis could be of much value as understanding the factors making other tissues are highly reliable to tumor growth or primary transformation. Simply stated, cancer cells may be strongly influenced by the surrounding muscle tissue in a manner that affects tumor suppression. In this study both hypotheses have been tested using simple in vitro and in vivo models.This study reports that myogenic cells exert pronounced effects upon co-culture with metastatic melanoma (B16-F10) or lung carcinoma (LLC1) cells including conversion to the myogenic lineage in vitro and in vivo, as well as inhibition of melanin production in melanoma cells coupled with cytotoxic and cytostatic effects. Additionally, biochemical examination of the proliferation and apoptotic cell markers showed that myogenic environment represses cell proliferation and provokes melanoma cell death when co-cultured with myogenic cells in two separate layers sharing the same medium. These results have been similarly extended to other metastatic cell lines as showed by Luis Lung carcinoma (LLC1) cells.On the other hand, no effect is seen with non-tumorigenic cells. Tumor suppression assays reveal that the muscle-mediated tumor suppressor effects do not generate resistant clones but function through the down-regulation of the transcription factor MiTF, a master regulator of melanocyte development and a melanoma oncogene. Our findings point to skeletal muscle as a source of therapeutic agents in the treatment of metastatic cancers.

Biography

Biography (Summary) Dr. Gomaa had her B.Sc. in Biology at the Faculty of Science, Cairo University. She fulfilled her masters between Cairo University and Panum Institute - Copenhagen University through a scholarship awarded from the European Union Community. She was then granted a Doctoral scholarship from the German Academic Exchange Services (DAAD) for undergoing her PhD between Ain Shams University, Cairo and the Medical School of the Technical University of Munich - Germany. She then followed her career path by completing four years of postdoctoral studies between Mount Sinai School of Medicine, NY - USA and the Faculty of Medicine, Marie Curie University (ParisVI), Paris - France. Currently she is an associate professor of Molecular and Cellular Biology at the Biotechnology Sector, Faculty of Pharmacy and Biotechnology � German University in Cairo.

Speaker
Iman Emam Omar Gomaa German University in Cairo (GUC), Egypt.

Abstract

We found out that a cell permeable dominant negative survivin (SurR9-C84A) competitively inhibited endogenous survivin and blocked the cell cycle at G1/S phase. Nanoencapsulation in mucoadhesive chitosan nanoparticles (CHNP) substantially increased its bioavailability and serum stability. We investigated the anti-cancer activity of alginate coated chitosan nanoparticles (CHNPs) encapsulating SurR9-C84A with aptamers targeting EpCAM and nucleolin. We incorporated three locked nucleic acid (LNA) modifications in each sequence in order to enhance the stability of these aptamers. Confocal microscopy revealed binding of the LNA-aptamers to their specific markers with high affinity. In colon cancer mouse xenograft model, the muco-adhesive LNA decorated nanoparticles showed 6-fold higher internalization and tumour specific uptake. A higher intensity of nanobullets was observed in both periphery and core of the multicellular tumour spheroids compared to non-targeted CHNP-SurR9-C84A. The nanoparticles were found to be the highly effective as they led to a 2.26 fold (p< 0.05) reduction at 24 h and a 4.95 fold reduction (p≤ 0.001) fold reduction in the spheroid size at 72 h. In conclusion, our nanoparticles have shown highly promising results and therefore deliver a new conduit towards the approach of cancer-targeted nanodelivery. Alkali burn is a frequently occurring ocular injury that resembles ocular inflammation caused by eye allergies, infection, and refractive surgeries. Our results confirmed that combination of SurR9-C84A with TSA worked in synergy to heal ocular injury and inflammations due to alkali burn and led to the regeneration of ocular tissue by increasing clathrin, claudin, survivin, and TGF-β and reversal of alkali burn by suppressing IL-1α and MMP-9 without inducing haze.

Biography

Professor Jagat R Kanwar is the Head and team leader of Nanomedicine-Laboratory of Immunology and Molecular Biomedical Research (NLIMBR), School of Medicine, Faculty of Health, Deakin University, Australia. Professor Kanwar has an international reputation and expertise in investigating fundamental and applied molecular signalling aspects of pathogenesis of cancer, chronic inflammation and neurodegenerative diseases, thereby, leading to the development of treatment strategies from bench to bedside. He has more than 150 research publications in high impact factor and peer reviewed international journals, 27 book chapters and 3 edited books. Prof Kanwar's research has generated several patents/PCTs with more than five licensed patents for commercialization to BioPharma industry. His group is currently working on drug discovery and nanomedicine for oral and systemic drug delivery of a range of natural bioactive and biomacromolecules (proteins/peptides, siRNAs and aptamers) for targeting survivin, HIF-1α and other apoptotic and inflammatory cell signalling molecules in cancer, chronic inflammation and neurodegenerative disorders. His research combines Immunology with state of the art and cutting edge techniques in Molecular Biology, Biochemistry, Nanobiotechnology and visualization to investigate the pathways in which key molecules are regulated in both normal and disease states. A number of in vitro human cell/tissue based co-culture models for cancers, microbial infections; autoimmune diseases; chronic inflammatory diseases (osteoarthritis, inflammatory bowel disease), gut health, neurodegeneration and immunomodulation have been developed by his group. Kanwar's main research objective is to understand and target the mechanisms involved at the molecular and sub cellular level which gives us an edge over the prevalent targeting techniques. He carries out both academic and commercial research projects and develops new approaches for the diagnosis, treatment, and nanomedicine based new generation delivery systems. His recent research focus on locked nucleic acid (LNA) LNA-modified aptamers conjugated

Speaker
Jagat R Kanwar Deakin University, Australia

Abstract

The success of nucleic acid therapeutics for disease treatment requires safe and effective gene transfer reagents. Hence, the search for more effective gene transfer reagents, or vectors, continues. Synthetic cationic lipids represent one of most active areas in the development of new non-viral vectors. Important considerations towards the development of new delivery agents are enhanced uptake efficiency, low toxicity and traceability. Our development and preliminary investigations of novel cationic glycol phospholipids, synthesized from chromophoric, rigid polyenoic C30:9 chains next to saturated flexible alkyl chains of variable lengths C12-20:0 lead to the first use of polyene-based lipids as siRNA transfer reagents. Furthermore, the chromophore associated with the polyene chain allowed in vitro tracking of the siRNA delivery using direct observation. The polyene lipoplexes were tracked both on a macroscopic and microscopic level as either a single-component or as a multi-component lipoplex formulation. Subsequently, we developed novel lipid analogues of a naturally occurring lipofuscin fluorophore, A2E. These A2E-based gene carriers, together with their associated red fluorescent emission allow us to visualize in vitro lipid/siRNA particle uptake and co-localization within cells.

Biography

Dr. Michael Pungente joined Weill Cornell Medical College in Qatar (WCMC-Q) as Assistant Professor of Organic Chemistry in 2003 and was promoted to Associate Professor in 2011. He received his Ph.D. in Organic Chemistry from the University of British Columbia (UBC) in 1997, and subsequently was Lecturer with the Department of Chemistry and then an Instructor with the Faculty of Pharmaceutical Sciences at UBC. Before joining WCMC-Q, Dr. Pungente held the position of Assistant Professor in the Department of Chemistry at Lakehead University in Thunder Bay, Ontario. His area of research includes the synthesis and characterization of novel lipids and polymers as potential non-viral gene therapy reagents.

Speaker
Michael Pungente Weill Cornell Medicine, Qatar

Abstract

The significant role of physiological dcEF in guiding cell movement during embryo development, cell differentiation, and wound healing has been demonstrated in several studies but the mechanism of cell electrotaxis is poorly understood. Additionally, there is no research addressing the question on the difference in electrotactic motility of cells representing various strategies of cell movement. To establish if cells representing different strategies of migration show a similar electrotactic reaction to external electric fields we constructed a unique experimental model which allowed for the investigation of electrotactic movement of cells of the same origin but representing different modes of cell migration: weakly adherent, spontaneously blebbing (BC) and lamellipodia forming (LC) WC256 cells. Both BC and LC sublines show robust cathodal migration in a physiological EF (1-3 V/cm). The directionality of cell movement was completely reversible upon reversing the field polarity. However, the full reversal of cell direction after the change of EF polarity was much faster in the case of BC (10 minutes) than LC cells (30 minutes). The proteomic composition of blebbing and lamellipodia forming sublines of WC 256 carcinosarcoma cells suggested that both sublines use a similar molecular machinery for cell movement and the different modes of migration resulted from differences in the regulation of activity of proteins involved in these processes rather than in differences in protein expression. Therefore, we investigated the distinct requirements for Rac, Cdc42 and Rho pathways and intracellular Ca2 in electrotaxis of WC256 sublines forming different types of cell protrusions. We found that Rac1 is required for directional movement of LC to a much greater extent than for BC, but Cdc42 and RhoA are more crucial for BC than for LC cells. The inhibition of ROCK did not affect electrotaxis of LC in contrast to BC cells. The results also showed that intracellular Ca2 is essential only for the electrotactic reaction of BC cells. Moreover, inhibition of MLCK and myosin II did not affect the electrotaxis of LC in contrast to BC cells. In conclusion, our results revealed that both lamellipodia and membrane blebs can efficiently drive electrotactic migration of WC 256 carcinosarcoma cells, however directional migration is mediated by different signalling pathways.

Biography

Prof. Zbigniew Madeja is a Professor of Cell Biology, Head of Department of Cell Biology and Dean of Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University in Krakow, Poland. He earned his B.Sc. (1985), Ph.D. (1991) and DSc (2003) in biology at Jagiellonian University. His current research is focused on: cell motility and cell-cell interactions among tumour cells and normal tissue cells, electrotaxis, chemotaxis and contact guidance of normal and tumour cells. Moreover he is involved in research on acceleration of wound healing, mass cultures of human skin keratinocytes, fibroblasts and melanocytes, application of electrochemistry to cell biology (elctrofusion, electroporation, cell electrophoretic separation) and analysis of the role of cell-substratum and cell-to-cell interactions in the regulation of cell physiology with special emphasis to the mutual interactions between factors determining cell adhesion and intercellular communication.

Speaker
Zbigniew Madeja Jagiellonian University, Poland

Abstract

BackgroundMutating Multi-drug resistant fecal bacteria (MRF), including gram-negative Escherichia coli and gram-positive Enterococcus faecium, have proven to be of prime concern to public health and food safety. Vancomycin-Resistant E. faecium (VRE) remain the leading cause of hospital-acquired infections with 10,000 hospitalized cases and 650 deaths each year in the U.S. Conjointly in this pathogenic spread, E. coli is implicated in millions of extraintestinal infections, resulting in more than 100,000 cases of sepsis and 40,000 sepsis-associated deaths. As such, an alternative natural antimicrobial that is nontoxic and sustainable was deemed of prime interest. Chitosan is emerging as a strong, natural antimicrobial that is considered safe for human health and have proven to be effective as an antibacterial against gram-negative strains and biofilms. Additionally, Zinc oxide (ZnO) is emerging as an effective nanocomponent for its potential to mitigate some gram-positive strains with regard to its selective toxicity. The objective of this study was to test the efficacy of a scalable nanoremediation on drug resistant fecal indicators and prove its mechanistic effects through Transmission Electron Microscopy (TEM).MethodsMulti-drug resistant (MDR) E. coli BAA 2471 and E. faecium 1449, wild type (WT) E. coli MCC 13 and E. faecium ATC 35667, and their respective co-cultures were grown to a concentration of ~5�105 CFU/mL in tryptic soy broth media. Nanoparticles utilized are Chitosan 3kDa (C1), Chitosan 50kDa (C2), Zinc Oxide (ZNP), a 1:1 mixture of C1 and ZNP (C1ZNP), and a 1:1 mixture of C2 and ZNP (C2ZNP). The minimum inhibitory concentration (MIC) test conforming to NCCLS Standards was performed using 96 well plates loaded with nanoremediants at a 2-fold dilution prior to the addition of 50 �L broth with bacteria growth. Trials (n>10) were averaged and represented in total nanotherapeutic concentration (mg/ml). The mechanistic effects of CZNPs on the fecal indicators were visualized through TEM, involving osmium tetroxide fixation. Bacteria were exposed for 24h before fixation to a concentration 65.5mg/ml of nanoparticles.ResultsC1 and C2 were found to only be effective against E. coli BAA 2471 (0.04 mg/ml) and were ineffective against all other bacteria cultures tested. Across trials, ZNP was found to be effective at varying concentrations, including an MIC of 5.208 and 6.25 mg/ml for trials against the co-cultured MDR and WT, but was found to be significantly more effective with the addition of C1 and C2. Against the co-cultures of MDR and WT bacteria, C1ZNP was effective at 0.781 and 2.083 mg/ml respectively, while C2ZNP was effective at a greater concentration of 1.302 and 3.125 mg/ml respectively. TEM visually confirmed chitosan acting as a transporter of ZnO in both Gram positive and negative cells, causing protrusions, lysis, and cell death. Observations of cellular structure, electron density, and cell wall appearance of MRF�s were markedly impacted by CZNPs compared to microbial control cells.ConclusionsSynergism of chitosan and ZnO proved to be inhibitive of MRF�s. This broad spectrum of activity has not been noted before. Low molecular weight chitosan, C1, was more impactful towards MRF�s than C2. TEM analyses demonstrated attachment and lysis of microbes at 24h past treatment. Conclusively, CZNPs inhibit MRF�s at concentrations lower than typical WT gut flora and holds promise as an effective in vivo intervention agent.

Biography

Speaker
Alya Limayem University of South Florida,UAE

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