Day2

  • Hokkaido University, Japan
  • Title:Prostaglandins in Teleost Ovulation: A Review of the Roles with a view to Comparison with Prostaglandins in Mammalian Ovulation.
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Abstract:
Ovulation, which is induced by the ovulatory luteinizing hormone (LH) surge in vertebrates, is a dynamic process that results in a discharge of one or more fertilizable oocytes from the ovarian follicle into the ovarian cavity or into the abdominal cavity. Follicle rupture is a core event of the ovulatory process and has been the subject of intensive investigation. Prostaglandins are well known to be central regulators of vertebrate ovulation. Studies addressing the role of prostaglandins in mammalian ovulation have established that they are involved in the processes of oocyte maturation and cumulus oocyte complex expansion. In contrast, despite the first indication of the role of prostaglandins in teleost ovulation appearing 40 years ago, the mechanistic background of their role has long been unknown. However, studies conducted on the teleost medaka over the past decade have provided valuable information. Emerging evidence indicates a critical role of prostaglandin E2 and its receptor subtype Ptger4b in the process of follicle rupture. In addition, our studies have revealed that activation of the melatonin/melatonin receptor system in the ovulating follicle is required for the prostaglandin E2-mediated follicle rupture process. In this talk, we summarize studies addressing the role of prostaglandins in teleost ovulation and describe recent advances. To help understand differences from and similarities to ovulation in mammalian species, the findings on the roles of prostaglandins in mammalian ovulation are discussed in parallel.
Biography:
*Takayuki Takahashi, Ph.D., Emeritus Professor of Hokkaido University, Japan
1979 Ph.D. in Zoology (Hokkaido University)
1980-1986 Research Associate, Protein Studies Lab at Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA
1987-1993 Assistant Professor, Faculty of Science, University of Tokyo, Japan
1993-2016 Professor, Faculty of Science, Hokkaido University, Japan
2016-2019 Research Professor, Hokkaido University, Japan
Research interests
My research activities are in the field of reproductive biology of vertebrates including fish. In particular, my laboratory has been closely associated with ovulation studies using a teleost medaka by cellular and molecular biological approaches. Our final goal is to get insights into the evolutional aspect of ovulation in the animal kingdom.
Achievements
We were the first to report that, using the teleost medaka, (i) MMPs and plasminogen activator/plasmin are involved in follicle rupture during ovulation (PNAS, 2005; Biol. Reprod., 2012, 2015). In addition, our studies showed that (ii) many ovulation-related-genes are induced by the surge of LH at ovulation (PLoS ONE, 2013; Biol. Reprod., 2014; MCE, 2017), (iii) activation of the prostaglandin E2 and the receptor system is required for follicle rupture (MCE, 2011, 2012, 2017; Biol. Reprod., 2016), and (iv) follicle rupture during ovulation is the process which proceeds under precise endocrine control (MCE, 2017; Reproduction, 2019; cells, 2019).
Award
In 2010, Prize from the Zoological Society of Japan
“Discovery of ovulatory proteases and elucidation of follicle rupture mechanism in teleost medaka”

  • University of Castilla-La Mancha , Spain
  • Title:Dlk Proteins Modulate The Osteogenic Differentiation of C3h10t1/2 Mesenchymal Cells
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Abstract:
DLK1 and DLK2 are transmembrane proteins known to be involved in the regulation of cell differentiation processes, such as adipogenesis, through the negative modulation of NOTCH signaling. Here, we examinate their possible roles in the osteogenic differentiation of mesenchymal C3H10T1/2 cells. We generated stable transfectants with plasmids that allow overexpress or diminish the expression of DLK1 or DLK2 proteins. Expression levels of these proteins were confirmed by using RT-qPCR and Western blot assays, and the activation of global NOTCH signaling was assessed by luciferase assays. We performed osteogenesis assays for 21 days and analyzed the mRNA expression of osteogenic markers, by using RT- qPCR assays, and the osteogenic capacity, by using the alkaline phosphatase staining method. Finally, we measured the phosphorylation levels of ERK1/2 MAPK, an essential kinase in osteogenesis, by using Western blot. Overexpression of DLK1, which inhibits NOTCH signaling, in C3H10T1/2 cells diminished the expression of osteogenic markers, alkaline phosphatase staining levels and ERK1/2 MAPK phosphorylation levels. Similarly, treatment with DAPT, a pharmacological inhibitor of NOTCH signaling, inhibited osteogenic differentiation. In contrast, overexpression of DLK2, or a lower expression of DLK1, promoted osteogenic differentiation by increasing the expression levels of osteogenic markers, alkaline phosphatase staining levels and ERK1/2 MAPK phosphorylation levels. On the other hand, low DLK2 expression did not bring out significant changes in the osteogenic process of these cells. Given these results, we concluded that DLK1 inhibits osteogenic differentiation of C3H10T1/2 cells, similarly to the NOTCH signaling inhibitor, DAPT, and inhibits ERK1/2 phosphorylation, whereas DLK2 enhances osteogenesis and activates ERK1/2 MAPK phosphorylation.
Biography:
I studied a degree in Pharmacy at University of Castilla-La Mancha, Spain. During those years, I got the chance to join several research teams and learn different laboratory skills thanks to the Research Initiation Program developed by the Faculty of Pharmacy. In 2016, when I finished my degree, I became a PhD student, got a grant and joined the research team of Biochemistry and Molecular Biology at the Faculty of Medicine, where I did my PhD Thesis, with honors, about the role of NOTCH receptors and DLK proteins in the osteogenic differentiation of C3H10T1/2 cells. These results have been presented in various national congresses and published in some articles in international journals. Now, I go on studying the role of these proteins in different processes related with cell proliferation and differentiation, such as adipogenesis.

  • University of Castilla-La Mancha , Spain
  • Title:NOTCH Receptors in Osteoblastogenic Differentiation.
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Abstract:
The purpose of this work is to analyse the role of NOTCH receptors on the osteoblastic differentiation of mesenchymal C3H10T1/2 cells. To reach this objective cell populations stably overexpressing each of the four NOTCH receptors were generated by transfection with a plasmid. Expression levels of Notch1-4 genes and its
target genes, Hes1 and Hey1, were assessed by RT-qPCR. The level of each NOTCH receptor in its overexpressing population was determined by Western blot. NOTCH activity was measured by luciferase assays. The osteoblastic differentiation capacity of each population was evaluated by the alkaline phosphatase staining method of induced cell cultures and the measurement of the expression levels of osteogenic differentiation markers in RNA samples obtained from differentiation assays.
The overexpression of a single NOTCH receptor produces an increase in the global NOTCH activity and modifies the expression of the other Notch and their target genes, Hes1 and Hey1. The generated transfectants showed different levels of osteogenic differentiation in in vitro assays. Alkaline phosphatase staining was more intense in cells overexpressing NOTCH3 or NOTCH2 and less intense in cells overexpressing NOTCH1 or NOTCH4. Populations overexpressing NOTCH1, NOTCH2 or NOTCH3 exhibit an increase in mRNA expression of markers compared with the control. Cells overexpressing NOTCH4 only showed a significative increase in Alpl marker expression.
Results indicate that the expression of Notch, Hes1 and Hey1 is interrelated and that the overexpression of the NOTCH1-3 receptors seems to have an inducing effect on the osteoblastic differentiation of C3H10T1/2 cells, while NOTCH4 overexpressing cells does not seem to modify the osteoblastic process.
Biography:
I pursued my bachelor’s degree in biotechnology in the Technical University of Madrid (2016/2020). During that period, I worked in plant molecular biology in the Centre of Plants’ Biotechnology and Genomics (CBGP). During 2020/2021 academic course I did my Maestry in Experimental Biomedicine. Since September 2020 I have been researching in cell differentiation in the Biochemistry Lab of the Medicine Faculty (UCLM).

  • University of Castilla-La Mancha , Spain
  • Title:High Expression Levels of DLK2 Protein in Human MDA-MB-231 Breast Cancer Cells Inhibits NOTCH1 Activation and Tumor Growth in Vivo.
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Abstract:
NOTCH receptor signaling has been implicated in the development of triple-negative breast cancer tumors. DLK2 protein, a non-canonical NOTCH signaling inhibitor has previously been shown to be involved in skin cancer. In this work, we studied whether different DLK2 expression levels affected the breast cancer features of MBA-MD-231 cells. We found that higher DLK2 expression levels led to stronger NOTCH1 inhibition. Cell cycle dynamics and apoptosis were affected by DLK2 in opposite directions, depending on the levels of NOTCH1 inhibition generated by the different levels of DLK2 expression. Only a strong inhibition of NOTCH1 by DLK2 led to a decrease in cell growth. The invasive properties of these cells were also affected. Low DLK2 expression levels led to an increase in invasiveness, whereas higher DLK2 expression levels did not significantly modify MBA-MD-231’s invasive properties. These alterations were associated to changes in the expression levels of cell adhesion proteins, such as N- and E-CADHERIN. DLK2 expression levels also affected some members of other cell signaling pathways, including ERK1/2 MAPK, AKT, and rpS6 kinases. Finally, MDA-MB-231 cells expressing high DLK2 levels were unable to generate tumors in vivo in a nude mice model. Our data support an important and complex role of DLK2 protein in the control of NOTCH signaling, tumor properties and growth dynamics of triple-negative breast cancer cells.
Biography:
I graduated in Pharmacy in 2000 from San Pablo CEU University (Spain). After obtaining a predoctoral fellowship, I defended my Diploma of Advanced Studies in Biochemistry and Molecular Biology at the Complutense University of Madrid and subsequently, my Doctoral Thesis on adipogenic differentiation, which obtained the highest grade, at the UCLM, Spain (2005). Within my scientific training, my stays abroad in prestigious laboratories, such as the FDA, Maryland and KMEB, Denmark, stand out. In 2008, I joined the Oncology Research Unit of the Albacete Hospital where I initiated a line of research on the study of DLK proteins in tumor processes.
Currently, I work as Assistant Professor and teach Biochemistry and Molecular Biology in the Pharmacy, Medicine and Biotechnology Degrees and I actively participate in several academic commissions (UCLM). During these years, I have directed several TFG, TFM, DEA and Doctoral Theses that have received the highest marks. I have published several articles in international journals, mainly related to the effect of DLK and NOTCH proteins in tumor growth and migration, as well as in adipogenic differentiation.

  • University of Castilla-La Mancha , Spain
  • Title:The Role of DLK1, an Inhibitory Non-Canonical Ligand of NOTCH Receptors, in the Osteoblastic Differentiation of Pre-Osteoblastic MC3T3-E1 Cell Line.
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Abstract:
We analyzed the role of DLK1 protein, an inhibitor of NOTCH receptors, in the osteogenic differentiation of MC3T3-E1 pre-osteoblastic cells, which lack endogenous Dlk1 expression. We have generated pools of cells stably overexpressing DLK1 protein by plasmid transfection. Dlk1 expression levels were analyzed by RT-qPCR and DLK1 protein levels by Western Blot. We performed osteogenic differentiation assays by treatment with culture medium supplemented with 10 mM β-glycerophosphate and 50 μg/ml ascorbic acid. The differentiation capacity of the DLK1-overexpressing cells was assessed by alkaline phosphatase staining and by the analysis of osteogenic differentiation markers by RT-qPCR. These assays were also performed using DAPT, a pharmacological inhibitor of NOTCH receptors. We have observed that the cells overexpressing Dlk1 showed a lower rate of osteogenic differentiation with respect to control cells, as shown by the lower degree of alkaline phosphatase staining and lower mRNA expression of some osteogenic markers, such as Alpl, Col1A1, Runx2 and Osteocalcin. The NOTCH receptor inhibitor, DAPT, also caused the decrease of both alkaline phosphatase staining and osteogenic markers in MC3T3-E1 cells treated with the osteogenic inducers. The results obtained indicate that Dlk1 overexpression or treatment with DAPT exert an inhibitory effect on the osteoblastic differentiation of MC3T3-E1 cells. This indicates that NOTCH signaling is required for osteogenesis of these cells and that DLK1 could exert its inhibitory effect on this process through inhibition of NOTCH signaling.
Biography:
I graduated in Biological Sciences at the University of Salamanca, Spain. My PhD Thesis was defended in April 1997 with honors. I started my postdoctoral stage at CBER (FDA, NIH Campus, Bethesda, MD, USA), where I applied my knowledge about yeast field to the study of the interaction of the mouse membrane protein DLK1 with the NOTCH1 receptor. In October 2000, I joined the Department of Biochemistry and Molecular Biology of the Faculty of Medicine of Albacete, UCLM, Spain. Now, I am working as a Full-Time Associate Professor, and I teach the first cycle of Biochemistry and Molecular Biology in the Degrees of Medicine, Pharmacy and Biotechnology. In my research work, I have mainly continued with the study of the function and mechanism of action of NOTCH receptors and DLK proteins in cell differentiation and cell proliferation. I have published several articles in international journals, whose papers have been presented in various national and international congresses. I have co-directed 3 PhD Theses, and I actively participate in several academic committees.

  • University hospital of Freiburg, Germany
  • Title:Enhanced AC133-Specific CAR T Cell Therapy Induces Durable Remission in Mice with Metastatic Small Cell Lung Cancer.
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Abstract:
Metastatic small cell lung cancer (SCLC) is not curable. While SCLC is initially sensitive to chemotherapy, remissions are short-lived. The relapse is induced by chemotherapy- selected tumor stem cells, which express the AC133 epitope of the CD133 stem cell marker. We studied the effectiveness of AC133-specific CAR T cells post-chemotherapy using human primary SCLC and an orthotopic xenograft mouse model. AC133-specific CAR T cells migrated to SCLC tumor lesions, reduced the tumor burden, and prolonged survival in a humanized orthotopic SCLC model, but were not able to entirely eliminate tumors. We identified CD73 and PD-L1 as immune-escape mechanisms and combined PD-1-inhibition and CD73-inhibition with CAR T cell treatment. This triple-immunotherapy induced cures in 25% of the mice, without signs of graft-versus-host disease or bone marrow failure. AC133+ cancer stem cells and PD-L1+CD73+ myeloid cells were detectable in primary human SCLC tissues, suggesting that patients may benefit from the triple-immunotherapy. We conclude that the combination of AC133-specific CAR T cells, anti-PD-1-antibody and CD73-inhibitor specifically eliminates chemoresistant tumor stem cells, overcomes SCLC-mediated T cell inhibition, and might induce long-term complete remission in an otherwise incurable disease.
Biography:
Current Position: Professor in Faculty Medical and Life Sciences University Furtwangen. Group Leader in Department of Hematology, Oncology and Stem Cell Transplantation, Medical Center – University of Freiburg, Germany.
Research: Dr. Taromi is molecular biologist; her research is concentrated on the development of novel CAR T cell-based approaches against SCLC. Dr. Taromi is conducting preclinical studies on the mechanisms of SCLC metastasis formation, inhibitory tumor microenvironment, stem cell-based resistance and T cell infiltration of solid tumors.

  • Danish Cancer Society Research Center, Denmark
  • Title:Compromising Plasma Membrane Repair in Cancer Cells.
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Abstract:
Injuries to the cell membrane of cancer cells, which are caused from invasive behavior, enhanced membrane dynamic and metabolic stress pose lethal threats to cancer cells. However, cancer cells cope by activating their plasma membrane repair system, which depends on mechanisms to remove damaged membrane by excision, internalization by endocytosis or reorganization of actin around the hole to reseal the wound. Proteins belonging to the annexin family are often found highly expressed in various cancer types and are characterized by their Ca2+-dependent binding to anionic phospholipids in the plasma membrane. Annexin family members share the ability to glue adjacent membranes together during wound healing but our recent results show that they play roles that are more specific in membrane repair by regulation of membrane excision, shedding, and induction of membrane curvature in cancer cells. Our findings open a novel avenue to target cancer cells by compromising annexin mediated plasma membrane repair. Here, novel molecular aspects of targeting the membrane repair system in cancer cells by phenothiazines, which alter plasma membrane properties and sensitize to membrane injury by inhibiting annexin-mediated repair will be presented.
Biography:
Dr. Jesper Nylandsted is Group Leader at the Danish Cancer Society Research Center in Copenhagen. His work focuses on alternative death pathways and repair mechanisms in cancer cells and novel approaches to target these processes. Using biophysical methods and live cell imaging, his group has revealed fundamental processes in cell death signalling and plasma membrane repair mechanisms of cancer cells.

  • University Kebangsaan Malaysia, Malaysia
  • Title:Cytotoxicity of Antimicrobial Peptides from the Skin Mucus of Anabas Testudineus on Breast Cancer Cell Lines.
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Abstract:
In this study, we characterised and evaluated the anticancer activity of the antimicrobial peptides (AMP)s excreted by Anabas testudineus. To induce AMPs production, A. testudineus was exposed to heat de-activated Pseudomonas aeruginosa for 10 days. The secreted mucus was then extracted and subjected to antibacterial activity testing by disc diffusion method to confirm the fish’s AMPs production. The crude mucus was able to inhibit P. Aeruginosa, and Escherichia coli (inhibition zone were 12 ± 0.43 mm and 10 ± 0.23 mm, respectively). The crude mucus was then screened against human breast cancer cell lines MCF7 and MDA-MB-231 with IC50 of 4.27 ± 0.15 mg/ml and 4.97 ± 0.25 mg/ml, respectively. After further fractionation, identification through peptide sequencing and anticancer screening, two peptides, designated as AtMP1 and AtMP2 (23 kDa and 20 kDa respectively) has been identified to inhibit MCF7 and MDA-MB-231 cell lines. These peptides were then synthetically produced and subjected to cytotoxic assay to prove its efficacy against breast cancer cell line. The IC50 for AtMP1against MCF7 and MDA-MB-231 were 8.25 ± 0.14 μg/ml and 9.35 ± 0.25 μg/ml respectively, while IC50 for AtMP2 were 5.89 ± 0.15 μg/ml and 6.97 ± 0.25 μg/ml respectively. The peptide did not inhibit the proliferation of HS27 normal cell line. This study provided new prospects in the development of highly effective and selective cancer therapeutics based on antimicrobial peptides from animal mucus.
Biography:
Dr. Mohd Shazrul Fazry graduated from The University of Queensland, Australia with a Ph.D. in Carcinogenesis. Since then, he has been working with potential local food product and natural resources to combat cancer.

  • University of Zurich, Switzerland
  • Title:Myoglobin, Expressed in Brown Adipose Tissue of Mice, Regulates the Content and Activity of Mitochondria and Lipid Droplets.
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Abstract:
The identification of novel physiological regulators that stimulate energy expenditure through brown adipose tissue (BAT) activity in substrate catalysis is of utmost importance to understand and treat metabolic diseases. Myoglobin (MB), known to store or transport oxygen in heart and skeletal muscles, has recently been found to bind fatty acids with physiological constants in its oxygenated form (i.e., MBO2). Here, we investigated the in vivo effect of MB expression on BAT activity. In particular, we studied mitochondrial function and lipid metabolism as essential determinants of energy expenditure in this tissue. We show in a MB-null (MBko) mouse model that MB expression in BAT impacts on the activity of brown adipocytes in a twofold manner: i) by elevating mitochondrial density plus maximal respiration capacity, and through that, by stimulating BAT oxidative metabolism along with the organelles` uncoupled respiration; and ii) by influencing the free fatty acids pool towards a palmitate-enriched composition and shifting the lipid droplet (LD) equilibrium towards higher counts of smaller droplets. These metabolic changes were accompanied by the up-regulated expression of thermogenesis markers UCP1, CIDEA, CIDEC, PGC1-α and PPAR-α in the BAT of MB wildtype (MBwt) mice. Along with the emergence of the “browning” BAT morphology, MBwt mice exhibited a leaner phenotype when compared to MBko littermates at 20 weeks of age. Our data shed novel insights into MB’s role in linking oxygen and lipid-based thermogenic metabolism. The findings suggest potential new strategies of targeting the MB pathway to treat metabolic disorders related to diminishing energy expenditure.
Biography:
Thomas A. Gorr:
1982-1988:Study of Biology, Philipps-University Marburg, Marburg, Germany;
1989-1993:PhD thesis in Protein Chemistry at the Max-Planck-Institute for Biochemistry,
Martinsried, Germany;
1994:Dr. rer. nat. (PhD), Ludwig-Maximilians-University, Munich, Germany;
1994-2004:postdoctoral stay in USA: a) UT Austin TX; b) Harvard Medical School;
2005-today:Leader of independent hypoxia/metabolism research group; Univ. Zurich;
2014:Habilitation (PD) in Comparative Physiology, Vetsuisse Faculty, University Zurich.

  • Universitas Brawijaya, Indonesia
  • Title:MiR-155–5p Predictive Role to Decelerate Foam Cell Atherosclerosis through CD36, VAV3, and SOCS1 Pathway.
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Abstract:
MicroRNAs (miRNAs) are noncoding RNA molecules that play a significant role in atherosclerosis pathogenesis through post-transcriptional regulation. In the present work, a bioinformatic analysis using TargetScan and miRdB databases was performed to identify the miRNAs targeting three genes involved in foam cell atherosclerosis (CD36, SOCS1, and Vav3). A total number of three hundred and sixty-seven miRNAs were recognized and only miR-155-5p was selected for further evaluation based on Venn analysis. Another objective of this study was to evaluate the biological process and regulatory network of miR-155-5p associated with foam cell atherosclerosis using DIANA, DAVID, Cytoscape, and STRING tools. Additionally, the comprehensive literature review was performed to prove the miR-155-5p function in foam cell atherosclerosis. miR-155-5p might be related with ox-LDL uptake and endocytosis in macrophage cell by targeting CD36 and Vav3 genes which was showed from the KEGG pathways hsa04979, hsa04560, hsa04810, and GO:0099632. Furthermore, miR-155-5p was also predicted to increase the cholesterol efflux from macrophage by inhibit SOCS1 expression based on KEGG pathway hsa04120. Eleven original studies were included in the review and strongly suggest the role of miR-155-5p in foam cell atherosclerosis inhibition.
Key words: miR-155-5p; foam cell; CD36; Vav3; SOCS1
Biography:
Ermin Rachmawati
Lecturer in Faculty of Medicine and Health Sciences UIN Maulana Malik Ibrahim Malang, Indonesia
Doctoral student in Faculty of Medicine Universitas Brawijaya, Malang Indonesia

  • National University of Colombia, Colombia
  • Title:Characteristics of PlasWD-40-1 and the HSP70-2/BiP/GRP78 Homolog PfHSP70-2 Proteins, their Possible Correlation, Role in Exporting Proteins Beyond its Shore
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Abstract:
Globally, efforts to advance malaria control and prevention have focused on two main areas of research: (1) Finding vaccine candidates to immunize populations and prevent posterior infections, (2) Finding novel and unknown molecules and fields where remain interesting questions unsolved. The intracellular malaria parasite Plasmodium falciparum organization, particular organelles as endoplasmic reticulum and domains of the infected erythrocytes affected by changes and modifications to evading the immune response, are important basic studies. P. falciparum export virulent proteins and built a complex cell modified to transport proteins traversing three concentric membranes or cellular compartments as; plasmatic membrane of the parasite, parasitophorous vacuolar membrane, and plasmatic membrane of infected erythrocyte getting alterations for own benefit and survive. Released merozoites ending the intraerythrocytic cycle after 48 hours of the invaded cell cause symptoms of disease, physiopathology and are highly immunogenic. Our group prepared monoclonal antibodies to study the event that occurs when merozoites egress from their host, this immunogen inoculated to generate the antibodies recognizing proteins localized in the Protein export compartment (PEC). PEC is a domain of the endoplasmic reticulum that could be associated with egress merozoites. However, before studying this event, it is necessary to know the identity of proteins localized in PEC. One of the proteins localized is named Pf68kDa, which is the first resident protein of PEC, identified and is recognized by a monoclonal antibody, mAb7.
We show identification of the 68 kDa antigen of P. falciparum. Recently works identified Pf68 kDa as two proteins by different methodological strategies, and the same mAb7: the first protein identified as a homolog of HSP70/BiP/GRP78, named PfHSP70-2 by affinity chromatography and mass spectrometry, and the second protein identified, Plasmodium WD40 repeat-containing protein-1 (PlasWD40-1) identified from a cDNA expression library screened with Mab7. From the plaques analyzed, two positive clones ultimately were identified. Thus, Mab7 not only recognizes PfHSP70-2 but also recognizes a protein expressed by the cloned DNA fragment (MK618679). Now, We know more about the nature of PEC, and not much about PlasWD40 and Pf68kDa is not precisely known. It is unknown if a common epitope between PfHSP70-2 and PlasWD40-1 is a coincidence or both proteins are interacting or regulating functions to directing activities in the endoplasmic reticulum to exporting proteins to the plasmatic membrane of P. falciparum-infected erythrocytes. The target from these two proteins and common epitope or each one separately, could to proposing new possible therapeutic strategies against malaria.
Biography:
Dr. Gladys Thalia Cortés obtained her Ph.D. in Biotechnology from Science Faculty at Universidad Nacional de Colombia (2020), Master Science degree at Universidad Javeriana (1995), and specialization training in immunochemistry and cellular immunology at National Institute of Health from Tokyo (Japan, 1986). Her primary field is Bacteriologist. Her studies in cell Biology of Plasmodium began with profesor Winograd E. at the Instituto Nacional de Salud (Bogotá), Wiser M., as consultor and co-investigator from Tulane University. She continued searching the cell Biology of Plasmodium at National University of Colombia (2005- ) in collaboration with Biofísica y Biología de membranas, Dr. Camacho M. at Centro Internacional de Física, Public Health Department at Faculty of Medicine. Her group published recently aspects related to, identification of antigen named Pf68 kDaPlasmodium falciparum resident protein of and specialized domain of the endoplasmic reticulum, or Plasmodium export compartment (PEC), PfHSP70-2 and (2020) and Plasmodium WD40 repeat-containing protein-1 (PlasWD40-1) identified from a cDNA expression library screened with Mab7 (2021), both proteins hypothesized as share a common epitope recognized by a same monoclonal antibody, mAb7 (doctoral thesis, 2019); besides, a study of release merozoites process from its host by photoconversion fluorescent compound to electron microscopy (2011), and characterization of proteins localized to a subcellular compartment associated with an alternate secretory pathway of the malaria parasite (2003). Currently, she is a Ph.D. researcher and professor of the Department of public health Faculty of Medicine at Universidad Nacional de Colombia.

  • Buckingham Browne & Nichols School, USA
  • Title:The Application of DNA Nucleotide Footprint Plotter and Peptide Visualizer to Coronavirus
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Abstract:
Direct visualization of the key features of coronavirus genomes and peptides can lead to a better understanding of this virus as well as to distinguish the mutant strains. The current study developed Java tools DNA nucleotide footprint plotter and peptide visualizer. DNA nucleotide footprint plotter makes it possible for straightforward visualization of the characteristics of viral genomes and recombination. Peptide visualizer is sensitive to changes in protein sequences and functions. The tools provide novel insights for biological studies that can contribute to breakthroughs in coronavirus diagnosis, treatment, and prevention.
Biography:
I am a junior in Buckingham Browne & Nichols school in Cambridge, Massachusetts, USA. I took biology and AP JAVA. The outbreak of coronavirus made me realize the importance to write tools to explore the genome and peptide characteristics of this virus. So I developed “DNA nucleotide footprint plotter” and “Peptide Visualizer”. The tools can directly display the key features of viral genomes and peptides and make intuitive visualization of different types of virus genomes and proteins.
In my leisure time, I spend a lot of time playing and writing video games, playing violin, participating robotics club and leading a rocketry club. I also enjoy crewing and scrolling on Charles river in Boston.

  • University of Tartu, Estonia
  • Title:open-source solutions for whole slide imaging: an overview and possible application.
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Abstract:
Digital pathology is rapidly evolving, transforming qualitative paradigms to quantitative ones, increasing histological examination objectivity. With a compound annual growth rate of the market over 10%, scanning devices with a higher slide capacity, speed and resolution become available. Not of less importance is software, varying from free basic viewers ending with advanced machine learning algorithms promising (semi) automatic diagnosis. To a significant extent, the advantages of computer-assisted image analysis may first be experienced via open-source solutions. The session provides an overview of existing free WSI-oriented solutions and examples of possible applications in teaching, scientific research, and pathologists’ daily practice.

Biography:
Pathologist, the founder of Estonian Digital Pathology Association. Co-developer of the first digital-pathology department in Estonia. Co-author of digital pathology solutions for medical and medical technician students in Estonia.

  • Jagiellonian University ,Poland
  • Title:Silencing of RIPK4 Inhibits NF-κB Activation and IL-8 Expression in TNF-α Stimulated Melanoma Cells
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Abstract:
Metastatic melanoma is an aggressive skin cancer that is notoriously resistant to current cancer therapies. Studies focused on the reciprocal interactions of melanoma and immune cells demonstrated that microenvironment-derived tumour necrosis factor (TNF-α) might induce dedifferentiation of the melanoma cells. Interleukin-8 (IL-8), a chemoattractant cytokine that has been demonstrated to positively influence tumour growth through autocrine and paracrine signalling is constitutively expressed in melanoma, however, it has remained not fully understand what factors elicit its upregulation since its expression can be regulated through a variety of mechanisms.
RIPK4 (Receptor-Interacting Protein Kinase 4) is a 91.6 kDa serine-threonine kinase belonging to the RIP family responsible for keratinocytes differentiation and stimulation of proinflammatory cytokine syntheses, such as CCL5, CXCL11 and IL8. RIPK4 is also involved in the regulation of the main signalling pathways in the cell, such as Wnt, NF-κB and acts as a “linker” between the PKC and NF-κB pathways. Constitutive activation of NF-κB pathway leads to the deregulation of gene transcription including IL-8 and plays a key role in the regulation of invasive properties and treatment resistance of melanoma.
Our studies show that RIPK4 level is manifold higher in melanoma cells than in normal melanocytes. Using interference siRNA technique we diminished the level of RIPK4 in melanoma cells and found that silencing of RIPK4 decrease activation of p65 and p-IKKα/β. Thus, we studied if IL-8 expression upon stimulation with TNF-α differs between melanoma cells with diminished level of RIPK4 expression and negative control (non-specific siRNA) by qRT-PCR and ELISA. We identified that IL-8 regulation by TNF-α in melanoma cell was mediated through RIPK4 and NF-κB activation.
The study was supported by the National Science Centre Grant number 2018/31/N/NZ3/02625 and 2018/31/B/N25/01423

Biography:
I’m a PhD student in the Department of Biophysics. In my research I study signalling pathways in melanoma cells, focusing on the role of RIPK4 kinase in the regulation of melanoma cell invasiveness. I’m principal investigator in project “Engagement of RIPK4 in RAF1/MEK/ERK pathway in melanoma cells” funding by National Science Centre. Moreover recently I finished my project “Involvement of RIPK4 kinase in PKC/NFκB pathway in melanoma progression”, Grant was funded by KNOW 2018/2019 to Faculty of Biochemistry, Biophysics, and Biotechnology of Jagiellonian University.
My publication:
Skalniak L, Smejda M, Cierniak A, Adamczyk A, Konieczny P, Madej E, Wolnicka-Głubisz
A. p38 but not p53 is responsible for UVA-induced MCPIP1 expression. Mech Ageing Dev. Volume 172, June 2018, Pages 96-106
Conference:
1)XLV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian University in Cracow, Zakopane, Poland, February 2018; The involvement of RIPK4 in the regulation of melanoma cell invasiveness (poster presentation)
2)IV Nationwide Biomedical Symposium Esculap, Lublin, Poland, December 2017; The receptor – interacting protein (RIP) kinase family as a new target in anticancer therapies (oral presentation)
3)17th Congress of the European Society for Photobiology, Pisa, Italy, September 2017; The role of p38/p53 in UVAinduced oxidative stress and MCPIP1 increase (author of the poster)
4)XLIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian University in Cracow, Zakopane, Poland, February 2017; The role of p53 in UVA- induced oxidative stress and MCPIP1 increase (poster presentation)

  • Instituto de Cardiologa Calle, Colombia
  • Title:Type 2 Myocardial Infarction in a Patient with Acute Abdomen Due to an Incarcerated Amyand’s Hernia
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Abstract
Background: Type 2 myocardial infarction (MIT2) is characterized by higher mortality rates compared to conventional type 1 infarction according to the European Society of Cardiology (ESC) in 2018. The purpose of this case is to identify appropriate therapeutic measures. A case of an Amyand’s Hernia that produced an MIT2 is described in this work.
Case Report: A 77-year-old male was admitted to our emergency department for acute abdominal pain in the right lower quadrant associated with the presence of an ipsilateral inguinal hernia with signs of peritoneal irritation, while complaining of chest pain. A positive troponin indicated the presence of myocardial infarction. A laparotomy was performed with the finding of an incarcerated right inguinoscrotal hernia that contained the gangrenous and perforated cecal appendix (Amyand hernia type 3). The treatment consisted of surgical correction of the hernia, an appendectomy, antibiotics and support in the intensive care unit with a positive outcome. The diagnosis of Amyand hernia type 3 was established intraoperatively, and by imaging, confirming the presence of an MIT2 according to the criteria of the fourth definition of ECS infarction.
Conclusion: In the surgical environment it is strange to find patients who present with acute abdominal pain and a myocardial infarction at the same time. It is necessary for the consultant to recognize these two entities to make a correct diagnosis and provide timely treatment to reduce any possibility of patient mortality
Biography:
I am Silvia Barbosa, physician at the Fundacion Cardioinfaltil in Bogota Colombia, I am part of the research group of general surgery and emergency with the aim of improving and promoting the scientific development of my country and society. We are a leading hospital in Latin America and it is through science that we can get more answers every day by sharing knowledge.
My interest is to share the complexity of the human being, how two or more

  • Lomonosov Moscow State University , Russia
  • Title:Antipodal Cells of the Embryo Sac of Wheat as a Unique Object to Study Plant Polytene Chromosomes
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Abstract:
Polytene chromosomes are interphase chromosomes consist of homologous chromatids, which are amplified through consecutive cycles of endoreduplication and conjugate together. Polytene chromosomes are formed in the nuclei of cells that require intense synthesis of substances in highly specialized tissues.
Antipodal cells of embryo sac of wheat is an example of plant cells with polytene chromosomes. Antipodal complex is located in the ovule between the nucellus and endosperm. Antipodal cells form a complex consisting of three cell layers – basal, which contacts with the conductive tissues, middle and apical, which contacts with the endosperm. After double fertilization antipodal cells produce substances necessary for the emerging endosperm coenocyte. Antipodal cells belong to the tissues that perform trophic and barrier functions between the maternal organism and the tissues formed after fertilization. So they play key role in development of endosperm and moreover are crucial for formation of endosperm in cultivated cereals, which are food for all the humanity.
The aim of our work was to study the morphology of the nuclei and giant polytene chromosomes of antipodal cells. The work was performed on total specimens of embryo sacs isolated from fixed wheat ovules. We used methods of light microscopy and transmission electron microscopy.
We measured DNA content in the nuclei of the antipodal cells and found that nuclear DNA content varies in the cells of the complex. The basal layer cells contain less DNA than apical layer cells. During the differentiation DNA content of nuclei increases.
Structure of antipodal polytene chromosome differ from structure of animal polytene chromosomes. Animal polytene chromosomes have disks and interdisks as a result of conjugation of chromatids along the entire length of the polytene chromosome. In regions of polytene chromosomes with the intensive synthesis of RNA, decondensation of chromatin occurs and puffs are formed. On the contrary antipodal cells are characterized by giant chromosomes in the form of bundles of threads and conjugation is found only in centromere regions. At the initial stages of the differentiation nuclei of antipodals have approximately the same size and nonsegregated polytene chromosomes. Then the nuclei of middle and apical layer cells increase in size, they have isolated individual chromosomes.
To conclude, the antipodal cells of the embryo sac in cereals are a unique object for solving the fundamental problems of cell biology and a convenient model for studying the structure of the nuclei and the stages of reforming polytene chromosomes during ontogenesis.

Biography:
Tatiana Doronina is a 2nd year PhD student in Lomonosov Moscow State University, Biology Faculty in Russia at the Department of Cell Biology and Histology. The field of interest is plant cell biology, polytene chromosomes and programmed cell death.
Publications:
• Doronina, T. V., Chaban, I. A., & Lazareva, E. M. 2019) Structural and Functional Features of the Wheat Embryo Sac’s Antipodal Cells during Differentiation. Russian Journal of Developmental Biology, 50(4), 194-208.
• Doronina, T. V., Chaban, I. A., & Lazareva, E. M. 2019. Structural reorganization of nuclei of wheat antipodal cells during programmed cell death. Biopolymers & Cell, 35(3), 206.

  • Alexandria University, Egypt
  • Title:The Impacts of Seasonal Variation on the Immune Status of Nile Tilapia Larvae and their Response to Different Immunostimulants Feed Additives
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Abstract
Few data are available on the thermal tolerance of Nile tilapia fish larvae in relation to their immune status and survival. The aims of this work were to evaluate the immune status of one day old Nile tilapia (Oreochromis niloticus) larval stage collected at the beginning (March), middle (August) and at the end (October) of hatching season through morphometric assessment of the larvae parameters including yolk sac diameter, body length and width as well as the expression of some immune-related genes (rag, sacs and tlr), inflammatory (il1b and il8) and stress related genes (hsp27, hsp70). Also, to compare the effect of three different immunostimulants (β-glucan, Vitamin C, and methionine /lysine amino acids mix) on the expression of the studied genes at two variant temperatures (23±1°C and 30±1°C) in experimental study for 21 days. The immune status of Nile tilapia is affected by thermal fluctuation throughout the hatching season reflected by altered yolk sac size, length, and expression of the immune and stress related genes of the larvae, the best performances was observed at the beginning of the hatching season (March). High temperature (30 oC) suppress immune and stress responses throughout downregulation of all the genes under study, mask any effects for the immunostimulants, increased mortality in fish larvae suggesting narrow thermal tolerance range for the larvae compared with the adult fish.
We recommend the use of amino acid mix as immunostimulant for Nile tilapia larvae, it reduce the mortality percentage and improve cellular response. Also the use of β- glucan should be prohibited during this developmental stage of larvae, it induced the highest mortality percentage.

Biography
I Abeer F. El-Nahas is a professor of Genetics and Genetic Engineering at the Faculty of Veterinary Medicine, Alexandria University, Egypt science 2012. I completed my practical work of Ph.D. at Hokkaido University, Japan. My research interests lie in the area of Cytogenetics, Molecular Genetics, Gene manipulation, Gene expression, Genetic Toxicology and immunogenetics. I am the Editor in Chief of Alexandria Journal of Veterinary Medicine and serving as editorial board members and reviewers of many well reputed journals.
Thirty three research students altogether awarded Master and Ph. D degrees under her direct supervision and he has over 30 publications in international peer-reviewed journals with citation over 300 times and her publication H-index is 11.
Abeer F. El-Nahas is the co-author of numerous articles published in prestigious journals, including: Fish & Shellfish Immunology, Environmental Science and Pollution Research, Gene, Basic and Clinical Pharmacology and Toxicology, Pharmaceutical Biology, Ecotoxicology and Environmental Safety, Oxidative Medicine and Cellular Longevity, and others

  • Volgograd Research Anti-plague , Russia
  • Title:Modification of the Francis Medium for the Conversion of Histoplasma Capsulatum to the Yeast-Like Growth Phase
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Abstract:
Histoplasmosis is a dangerous deep systemic mycosis that occurs in people even with preserved immune status, which can occur as respiratory distress syndrome, tuberculosis and other deseases. The importance of verification of diagnosis is associated with the vital necessity of prescribing etiotropic antimycotic therapy for patients. The causative agent of histoplasmosis is the fungus Histoplasma capsulatum which exists in two phases – the mycelial (in the external environment) and the tissue (in the macroorganism). Proof of micromycete dimorphism is the basis for laboratory diagnostics of histoplasmosis, however, there is a problem of conversion of the pathogen into the tissue (yeast) phase on nutrient media in vitro.  The Francis medium is most known for this purpose, but in our studies FT-agar, intended for the cultivation of a special whimsicality to growth conditions tularemia microbe, enriched with fermented and defibrinated blood, D-glucose, sulfur-containing amino acids and vitamins complex proved to be  significantly more effective.  The number of colony-forming units on the FT-based Frensis medium compared with Francis medium or  FT-agar  without the appropriate additives in all the observations was significantly higher (p<0,01-0,001). The results obtained allow us to recommend this environment for practical use. These studies are confirmed by a Patent (RU 2 704 278 C1).

Biography:
Dr. Novitskaya I. V. has graduated from Volgograd State Medical University, Russia. For a long time, she has been working at the Volgograd research anti-plague Institute as the head of the Department and an associate Professor of the Faculty of molecular biology and genetics of the  medical University. Her research interests include Mycology, cell biology, biotechnology, diagnostics of dangerous and opportunistic infections. She is the author of more than 100 scientific papers, including Patents and Author’s certificates.

 

  • Dept RDDM , France
  • Title:Redox Properties of Mono- and Tri-Cyclic Cyanoenones Correlated with their Efficacy as Inducers of a Cytoprotective Phase 2 Enzyme and as Protectors against Inflammation: Potency Ranking
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Abstract:
Induction of the Phase 2 enzymes is a major strategy in the chemoprotection against cancer. Inducers belong to nine different chemical classes. In this contribution we found that a measure of the tendency of thirty plant phenylpropenoids and synthetic analogues to release electrons correlates linearly with their potency in inducing the activity of NAD(P)H:quinone reductase (NQO1), a prototypic Phase 2 cancer protective enzyme.  The tendency to release electrons was determined by the energy of the highest occupied molecular orbital (EHOMO), calculated by simple quantum mechanical methods. The correlations observed establish a clear conclusion: the smaller the absolute Energy of the Highest Occupied Molecular Orbital (EHOMO) of an agent, the greater its inducer potency, (i.e., the lower its oxidation potential, the stronger its electron donor property and the greater its inducer potency). The finding of this redox ranking of the inducers demonstrates that it is possible to predictively control the genetic expression of an enzymatic defense against cancer by xenobiotics via one physical parameter, their EHOMO.

  • Colombiana de Trasplantes ,Colombia
  • Title:Hand-Assisted Laparoscopic Nephrectomy: Evaluation of the Learning Curve
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Abstract:
Background. Hand-assisted laparoscopic donor nephrectomy (HALDN) has rapidly become the best alternative to open nephrectomy for living kidney donation. As more centers continue to adopt the laparoscopic technique, the safety of the initial transplants must be ensured while ascending the learning curve (LC). This study looks to determine the safety of HALDN and to describe the results of the LC in our center.
Methods. We conducted a retrospective review of 500 HALDNs performed in our center from July 2003 to July 2017. We analyzed demographic and perioperative characteristics and complications during the first postoperative month. We divided HALDNs into 2 groups: before and after completing the LC (50 nephrectomies). For each group, we assessed operating room time, estimated blood loss, length of stay, and complication and conversion rates.
Results. A total of 500 HALDNs were performed in the study period. Of those, 454 were analyzed in the 2 groups. The median operating room time was 2 hours, length of stay was 2 days, and blood loss was 50 cc. The overall rate of complication was 6.8%. There were significant differences between the 2 groups in operating time, blood loss, and length of stay (P < .05). No differences were found in terms of complication (P 1⁄4 .42) and con- version (P 1⁄4 .28) rates.
Conclusion. There was a significant decrease in operating time, blood loss, and length of stay in patients who underwent laparoscopic donor nephrectomy by an experienced lapa- roscopist. However, no differences were found in complication and conversion rates, which suggests that improvement in surgical training can be accomplished without altering the donor safety.

  • USA
  • Title:Analysis of Glycosylation in Monoclonal Antibodies
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Abstract:
Monoclonal antibodies (MAbs) are rapidly growing class of therapeutic molecules in biopharmaceuticals. More than 80 therapeutic antibodies have been approved by the FDA in the last 30 years for different indications ranging from oncology to autoimmune diseases to respiratory diseases and the number is increasing year by year. Most of the current therapeutic antibodies are immunoglobulin G (IgG) with glycosylation constituting around 3% of the total mass of the molecule. Understanding the impact of glycosylation and close monitoring is critical for monoclonal antibodies and fusion proteins development as therapeutic molecule. Different forms of glycan including sialic acid (NANA/NGNA), galactose, mannose and fucose influence safety, efficacy and pharmacodynamics/pharmacokinetic property (PD/PK) of the therapeutic monoclonal antibodies and fusion proteins. This presentation will highlight the influence of different glycan variants on the drug’s behaviour in the body and draw attention to commonly employed analytical techniques to analyse therapeutic molecules and determine and quantify glycan composition, structure and glycosylation site in them.

Biography:
Dr Harleen Kaur has been in the pharmaceutical industry for almost 8 years and most recently led the analytics and drug product tech transfer projects for two biologics products while working at AstraZeneca, USA. Prior to this, she worked in R&D division of Fujitsu Asia Pte Ltd in Singapore where she worked on aptamer development and played an integral role in identifying and purifying aptamers against different protein targets in collaboration with National University of Singapore, Agency of Science Technology and Research Singapore, and Japanese diagnostic enterprise Sysmex. In her current role, Dr Kaur is leading a team of analytical scientists at Aurobindo Biologics and her responsibilities include the method development, method qualification and method transfer for different biosimilar products. Dr Kaur completed her PhD in chemical and biomolecular engineering department at National University of Singapore.

 

 

  • Johns Hopkins University School of Medicine,USA
  • Title:Use of Contact Lenses to Optimize Optical Coherence Tomography Scans of the Optic Nerve in Open Angle Glaucoma Suspects or Patients with Open Angle Glaucoma with High Myopia
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Abstract:
Patients with myopia are at increased risk for the development of glaucoma. The inability to correct for axial length on spectral domain OCT (SD-OCT) imaging translates into a lower signal strength and scan reliability in patients with high myopia. We evaluated the effectiveness of a contact lens to increase the signal strength, assess optic nerve dimensions and nerve fiber layer thickness using SD-OCT in patients with glaucoma or who are glaucoma suspects with high axial myopia.
Methods- A single-center, prospective, interventional study of patients with axial lengths over 25.5 mm with a diagnosis of glaucoma or glaucoma suspect. The optic nerve cube 200 x 200 scan using the Cirrus SD-OCT 400 was taken first without and then repeated after the placement of the contact lens. The primary outcome measure was the change in the average nerve fiber layer thickness before and after use of the contact lens. Secondary outcome measures included the change in cup volume, disc area, and rim area.
Results- Twelve patients were recruited (20 eyes) and the average axial length was 27.06 mm and the average signal strength interval increased by 1.73 (p= 0.001). With the use of a contact lens, the average nerve fiber layer thickness was significantly thicker. None of the changes in the secondary outcome measures were significant: rim area, cup volume, or disc area. Conclusions- Based on our data, the use of a contact lens statistically improved the signal strength and average nerve fiber layer thickness of the SD-OCT scan. The ability to accurately capture the perimeter of the optic disc can be limited in the setting of peripapillary atrophy, which was present in all but two subjects. Future studies with a larger number of subjects and a wider range of axial myopia to discern if contact lens correction has a greater effect on the highest axial lengths are needed.

Biography:
Meghan K. Berkenstock: Dr. Berkenstock has become an emerging leader in the field of ocular immunology. As an Assistant Professor of Ophthalmology at the Wilmer Eye Institute of the Johns Hopkins School of Medicine, her research focuses on quality improvement and identifying and treating ocular side effects of oncologic immunotherapy medications.

  • Immune-Oncological Center Cologne,Germany
  • Title:Cancer Vaccines Modified by Virus Infection: Concept, Mechanism of Function and Clinical Results
  • Time :

Abstract:
Biological therapies such as immunotherapy and oncolytic virotherapy are physiological and well tolerated by cancer patients. The combination of cancer vaccines with oncolytic viruses is a powerful concept. Two types of autologous cancer vaccines will be described which are modified by infection with the avian Newcastle disease virus. They instruct the immune system about relevant cancer targets (tumor antigens, TAs) and contain signals for innate immunity activation. Viral molecular patterns such as S’-PPP RNA and hemagglutininneuraminidase (HN) protein initiate early inflammatory defense reactions which contribute to activation of antigen-presenting cells (APCs) and to costimulation of T cells. The concommitant stimulation of TA-specific CD4+ and CD8+ T cells occurs in T-APC clusters. These generate cancer-reactive cytotoxic T lymphocytes (CTLs) and T cell mediated long-term immunological memory. Results from pre-clinical and clinical studies will be presented.

Biography:
Volker Schirrmacher is Scientific Director of Tumorimmunology at aAZK, Cologne, Germany. He finished his studies of biochemistry with a PhD in immunology. After 5 years post-doc time in Stockholm, Sweden and London, UK, he became Full Professor and Head of Division of Cellular lmmunology at the German Cancer Research Center, Heidelberg, Germany. This position was hold for 32 years until official retirement in 2008. His scientific oevre covers more than 400 peer-reviewed publications and several books. He was awarded the Aronson Price, the Meyenburg Price and the German Cancer Award.

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