12^th International Conference on ^{Genomics and Molecular Biology} April 15-17, 2019 Berlin, Germany

Biography:

Yanyan Zhao is director of Department of Clinical Genetics in Shengjing Hospital of China Medical University. She was completed MD in 1984 and PhD in 1996 at China Medical University. She did postdoctoral studies in Department of Cell Biology of State University of New York. Over the past decades she has focused on molecular genetics of cardiovascular disease and prenatal diagnosis of Genetic diseased, and published more than 50 papers.

Abstract:

Nuclear receptor-binding SET domain-containing protein 1 (Nsd1) is a histone lysine methyltransferase, and the effect of NSD1 in response to oxidative stress in cardiac muscle cells remains poorly understood. Current study demonstrated that the protein and mRNA levels of NSD1 was reduced by H2O2 in H9C2 cells. We analyzed the promoter of NSD1 gene, predicted an NK2 transcription factor related locus 5 (Nkx2.5) binding element (NKE) at position -412/-406 in the NSD1 promoter, and confirmed it by an electrophoresis mobility shift assay and a chromatin immunoprecipitation assay. Luciferase activity analysis indicated H2O2 decreased the p646-luc promoter activity, and Nkx2.5-specific siRNA weakened H2O2 inhibition on the p646-luc promoter. However, H2O2 did not affected mutant NKE promoter p646-luc-mut compared with wild p646-luc promoter.  Furthermore, overexpression and depletion of Nkx2.5 led to the increase and decrease of NSD1 protein and mRNA levels in H9C2 cells. In addition, Nkx2.5 was downregulated by H2O2. Therefore we hypothesized that oxidative stress reduced NSD1 expression through suppression of Nkx2.5, and we going to focus on the alteration of histone methylation of NSD in response oxidative stress in future study.

Biography:

Maria Krzakowa works as Professor Senior at Adam Mickiewicz University in Poznań, Poland. Some years ago she did one- year training as Post Doc at the University of California Davis. She learned there enzymes detection by horizontal gel electrophoresis. At first, her scientific activity was concerned about genetic variation of natural populations of Bryophytes (different species of Hepatics and Mosses) and later on forest trees: Scots pine (Pinus sylvestris), beech (Fagus sylvatica) and ash-tree (Fraxinus excelsior). In the meantime she developed investigations on grasses, for example Apera spica-venti and Alopecurus myosuroides. Her main achievement was the first description of dimeric peroxidise in reed (Phragmites australis). It was some kind of discovery, as dimeric peroxidise was known earlier only from the rice (Oryza sativa). Now, she is working on European collection of Italian and Westerwolds ryegrasses (Lolium spp.) considering biochemical and molecular polymorphism of these important forage grasses.

Abstract:

Lolium westerwoldicum is an important forage grass originated from the Netherlands. It has been selected by farmers of Westerwolde from local rapid growing Italian ryegrass types within the populations of local variety. Westerwolths ryegrass is the shortest-life form of all annual ryegrasses. Genetic examination of this interesting species is very limited. One-month seedlings of L. westerwoldicum cultivars originated from different European countries: France (Barcomet, Gipsyl, Suxyl and Melword), Germany (Duccado, Grasser and Nerissa), Belgium (Mendoza), Netherlands (Bartimo and Gepetto) and Poland (Mowester) were treated as populations and investigated in terms of the two dimeric enzyme systems: phosphoglucose isomerase (PGI; EC 5.3.1.9) and diaforase (DIA; EC 1.6.4.3). From each cultivar, minimum 30 plants were examined in 11% starch gel (SIGMA), prepared on the basis of lithium-boric buffer system. Electrophoretically detected phenotypes were created by four alleles of one locus in PGI and two alleles in one locus in DIA were used to calculate the genetic parameters like heterozygosities (He and Ho)and polymorphic index (Pg). When all populations were compared according to PGI allozymes frequency, all populations show high polymorphism from Pg=0.53 for Mowester till Pg=0.81 for Gepetto. Polymorphic indices calculated for DIA are fluctuating from PG=050 for Bartimo till Pg=065 for Ducado.

All populations were compared on the basis of alleles frequency using Unweighted Pair Groups Method with Arithmetic Mean (UPGMA) and illustrated by dendrograms constructed for both: PGI and DIA enzyme systems. Dendrogram constructed for PGI allozymes shows two groups of populations: one composed of two cultivars: Mendosa (Belgium) and Melword (France) and the second group composed of 8 varieties. Population Bartimo from Holland is visibly separated. Dendrogram constructed for DIA on the basis of alleles frequency shows three groups: two composed of three populations and one of five populations. The first is formed by Bartimo from Holland as well as Suxyl and Gipsyl from France). The second three-species group is composed of Meword (France), Mowster (Poland) and Ducado (Germany). It is visible that connections between populations do not indicate regional similarity.

Biography:

Wen Qiu is currently an associate professor at Department of Immunology of Nanjing Medical University. He is exploring roles and mechanisms of inflammatory factors in glioma cell proliferation and migration. These include signal transduction, microRNA regulation, transcriptional factor regulation. He is also exploring the effects of post-transcriptional regulation such as ubiquitination and acetylation on the activation of signaling molecules and transcription factors

Abstract:

Interleukin 17 (IL‑17), as a pro-inflammatory cytokine, is up-regulated in the sera and tumor tissues of glioma patients; however the effects of IL-17 on glioma proliferation and migration remain unclear. In this study, the roles of IL-17 in the proliferation and migration of glioma cells and their potential mechanisms were determined. The results showed that IL-17 could not only enhance the proliferation and migration of cultured glioma cells (in vitro), but also promote the tumor formation of glioma cells in BALB/c nude mice (in vivo). Mechanical exploration revealed that IL-17 stimulation could increase the phosphorylation levels of Akt1 and NF-κB-p65 in glioma cells, and knockdown or inhibition of PI3K, Akt1 and NF-κB-p65 could also reduce the IL-17-induced proliferation and migration of the glioma cells. Moreover, PI3K/Akt1 was the upstream regulator of NF-κB-p65 activation in IL-17-incubated glioma cells. Furthermore, the inhibition of PI3K, Akt1 and NF-κB-p65 markedly suppressed the tumor formation of glioma cells induced by IL-17. Together, these data indicate that IL-17 can promote the proliferation and migration of glioma cells via PI3K/Akt1/NF-κB-p65 activation, and these findings might provide a new insight into glioma pathogenesis.

Biography:

Yulia Einav has completed her PhD in Tel Aviv University. She works at Holon Institute of Technology (Israel). Currently she has published about 25 papers and book chapters in reputed journals and textbooks. In addition, she serves as Dean of Students of her institution.

Abstract:

The cardiac ryanodine receptor RyR2 is a calcium release channel present in the sarcoplasmic reticulum, which plays a major role in regulation of Ca2+ homeostasis in the heart. We discovered a novel RyR2 missense mutation, G3118R, with recessive co-segregation, in a large family presenting with cardiac arrest and ventricular fibrillation phenotype. Interestingly, and unlike the other clinically relevant mutations in this gene, which are dominantly inherited, the clinically affected individuals in this family are homozygous for the mutation, whereas the heterozygous family members are almost unaffected. Moreover, the mechanism by which this mutation determines this unusual clinical phenotype is unknown. The aims of this study were to perform functional and structural analysis of G3118R RyR2 in order to investigate the mechanism of this mutation's pathogenicity. We inserted the mutation into HEK293 cells and performed functional studies, including measurements of changes in intracellular Ca2+ levels and the monitoring of the endoplasmic reticulum Ca2+ dynamics. G3118R causes suppression of function of the channel, which is by far less described mechanism caused by most RyR2 mutations. Next, we used a computational model to study the changes in the stability and the flexibility of RyR2 protein.  We found that G3118R, a peripheral mutation located in a region, whose function is unknown, causes a major allosteric effect on the channel pore region of two out of the four RyR2 monomers, and that this effect accumulates in a dose-dependent manner.

Biography:

Mariom is a PhD student at ‘The University of Tokyo’ and an assistant professor at ‘Bangladesh Agricultural University’. Her research interests lie in the area of molecular biology and biotechnology. Her work now focuses on molecular mechanisms underlying the formation of pearl sac and pearl because increased understanding towards producing a high quality lustrous pearl is required to further improve the effectiveness of pearl culture technique. She earned a bachelor of science in ‘Fisheries’ and a master of science in ‘Fisheries Biology and Genetics’ from ‘Bangladesh Agricultural University’.

Abstract:

In pearl farming, a piece of mantle tissues from a donor oyster is implanted into a host oyster along with an inorganic nucleus. The outer epithelial cells of the graft proliferate and form a pearl sac which secretes various shell matrix proteins (SMPs) to form a pearl surrounding the nucleus by a complex physiological process which has not been well-understood yet. Using an RNA-seq approach, here we aimed to unravel the expression of the key genes involved in pearl biomineralization at different stages. During grafting experiments for three months, we collected nine samples (donor mantle epithelial cells, donor mantle pallium, donor mantle pallium on grafting, and mantle pallium each from the host at 24 hr, 48 hr, 1 week, 2 week, 1 month, and 3 month post grafting). The pearl sac was developed by two weeks after transplantation. For the first time, we identified some stem cell marker genes differentially expressed during pearl sac formation. PCA analysis on 192 biomineralization-related genes showed clearly different expression profiles between before and after 1 week after grafting. The expression profiles of these genes demonstrated that prismatic layer forming SMPs were first up-regulated and then gradually down-regulated, indicating their involvement in the onset of pearl mineralization. Most of the nacreous layer forming SMPs were up-regulated after the formation of pearl sac with the highest expression at 1 month, suggesting the completion of nacreous layer formation. These findings provide valuable information in understanding the molecular mechanism of pearl formation in P. fucata.

Biography:

Anusha Patel has completed her Graduation in 2008; Masters of Pharmacy at Kings College London and Postgraduate training in various hospitals around London. Following obtaining a Diploma in Medicines Management from the London School of Pharmacy, she has specialised in Trauma and Orthopaedics Pharmacy at UHL for a number of years. She then took on a homecare role at Kettering which has evolved into the current position. She Chairs the Regional Homecare Group for the East Midlands, and is a member of the National Homecare Medicines Committee (NHMC). She also works with the East Midlands Procurement Collaborative to assist in tenders for homecare contracts.            

Abstract:

Homecare is a rapidly growing market with a value circa £1.5bn. Approximately 250,000 patients are receiving medicines via the homecare route. Biosimilars offer an exciting future: Opportunity to include more value added services, improved medicine administration (e.g. injectable medicines) through nurse support and patient education, improved patient compliance through offering help lines and checking product utilisation, improved product quality by enhanced supply chain, cold chain management, stock rotation, etc, improved track and trace systems, enhanced distribution models. Switching process at KGH-Currently a total of 100 patients on therapy via the homecare route. Rheumatology switched first, then dermatology, new patients commenced Benepali 21^st march, existing patients to switch to biosimilar as a phased process from 1^st April. Actions to take: discuss possibility of using Benepali with rheumatology team at MDT meeting, identify appropriate patients, notify patients beforehand-leaflet, clinic consultation to offer switch to existing patients, consultant/nurse to notify secretary of switch and to prepare a prescription (No registration forms needed), homecare pharmacist assistance in producing prescriptions, prescription processed by pharmacy and sent to homecare provider, homecare provider receive prescription, update patient account of switching to Benepali, homecare provider to contact patient, perform a stock check and supply “Benepali Patient Kit” on first delivery. Negotiations with the CCG-Gain share with CCG Gain share further between pharmacy and rheumatology department. Savings are to be used to recruit another nurse for department to release consultant nurse led telephone consultations for those patients who need them. Barriers to overcome were mapped out beforehand and a savings tracker was monitored to manage finances. Homecare providers are in a prime position to assist with switching as we can see in the preaparations for adalimuumab biolsimilar and trusts should work with them to ensure seamless transfer.

Biography:

Wanjie Sun is a Senior Mathematical Statistician at CDER of FDA. Prior to joining FDA, she worked at GWU as a Lead Research Scientist and a PI/co-PI for over ten years, and she also worked in pharmaceutical industry for a couple of years. She has obtained her PhD in Biostatistics at GWU. She has over 40 publications in statistical, medical, and pharmacy journals.  Her research interests are in equivalence, non-inferiority, causal inference, study design, and missing data.  

Abstract:

In equivalence, biosimilar, and non-inferiority studies, intent-to-treat (ITT) analysis tends to make the two treatments look similar, thereby is generally anti-conservative. The Per Protocol (PP) analysis based on completers and compliers is more able to reflect treatment differences and is usually preferred in equivalence assessment. In clinical endpoint bioequivalence studies, the current primary analysis for assessing equivalence between a generic and an innovator product is usually based on the observed per-protocol (PP) population. However, missing data and non-compliance are post-randomization intercurrent events and may introduce selection bias. Therefore, PP analysis is generally not causal. The FDA Missing Data Working Group (2016) recommended using “causal estimands of primary interest.” In this paper, we propose a principal stratification causal framework and co-primary causal estimands to test equivalence, which was one of the approaches recommended by the recently published ICH E9 (R1) addendum to address intercurrent events. We identify three conditions under which the current PP estimator is unbiased for one of the proposed co-primary causal estimands-the “Survivor Average Causal Effect” (SACE) estimand.  Simulation shows that when these three conditions are not met, the PP estimator is biased, and may inflate Type 1 error and/or change power. We also propose a tipping point sensitivity analysis to evaluate the robustness of the current PP estimator in testing equivalence when the sensitivity parameters deviate from the three identified conditions, but stay within a clinically meaningful range. Our work is the first causal equivalence assessment in equivalence studies with intercurrent events and can be applied to comparative biosimilar clinical trials and non-inferiority trials.

Recent Publications

1. Lou Y, Jones M P and Sun W (2018) Estimation of causal effects in clinical endpoint bioequivalence studies in the presence of intercurrent events: noncompliance and missing data. Journal of biopharmaceutical statistics 1-23.

2. LaVange L M and Permutt T (2016) A regulatory perspective on missing data in the aftermath of the NRC report. Statistics in Medicine 35(17):2853-2864.

3. Permutt T (2016) A taxonomy of estimands for regulatory clinical trials with discontinuations. Statistics in Medicine 35(17):2865-2875.

4. Permutt T (2016) Sensitivity analysis for missing data in regulatory submissions. Statistics in Medicine 35(17):2876-2879.

5. Sun W, Zhou L, Grosser S and Kim C (2016) A meta-analysis of missing data and non-compliance data in clinical endpoint bioequivalence studies. Statistics in Biopharmaceutical Research 8(3):334-344.

Biography:

Rufina Soomro completed her diploma in medical education (AKU), FACS(USA). She is working as Professor of Surgery at Liquate National Hospital and Medical College Karachi and She is as a chairperson of Postgraduate medical education committee of the same institute. She did Breast surgery training from Dublin, Ireland, and currently running one of the busiest breast services in the country. Has keen interest in research and did a research course from Harvard University. She is serving College and Physicians of Pakistan as an Examiner in G. Surgery, ATLS course Director / Instructor and Secretary of National Surgery faculty.

Abstract:

Introduction: Breast cancer is the commonest cancer in women worldwide and represents a highly heterogeneous group of tumours particularly in terms of molecular features, prognosis and response to therapy. Breast cancer molecular classification can predict the prognosis of breast cancer in terms of recurrence and help and guide us regarding the treatment decision about systemic therapy. Breast carcinomas may be stratified into subtypes similar to those defined by Gene expression profiling using a panel of immune-histochemical (IHC) markers. Routine IHC evaluations of breast cancers may, therefore, provide a reasonable alternative to costly genetic assays especially in under-resourced healthcare systems. The purpose of this study is to investigate the prevalence of molecular subtypes and correlate it to clinic-pathological features.

Methods: From 2005 to 2017 total of 4847 Breast cancer patients, in whom complete information was available to classify them into luminal subtypes were retrieved and classified into intrinsic subtypes and patients information in each type was collected about age, tumour size, stage, grade and nodal status.

Results: In luminal classification, a highly significant difference was found in mean age (p<0.001) tumour size (p<0.001), grade, metastasis and Ki67. The statistical significance of Her 2 positive and triple negative was found with stage, grade, metastasis and Ki67.

Conclusions: IHC assignment into Luminal subtypes is clinically informative in our patients and routinely using this in our practice could identify patients that may need a more aggressive treatment to reduce the likelihood of recurrences.

Biography:

Waraporn Kraitavin is a Ph.D. student in Aquatic Molecular Biology and Biotechnology at the University of Tokyo, Japan. She worked as a Nutritionist at Cargill Meats (Thailand) Limited for 1 and a half years and she has completed her Marter’s degree and Bachelor’s degree from Kasetsart University, Thailand

Abstract:

Understanding the mechanism of temperature adaptation is crucial for cold-freshwater fish in order to cope with the recent global warming, especially yamame (Oncorhynchus masou), which is an important aquaculture species belonging to the family Salmonidae. The aim of this study is to understand the adaptive response of high-temperature tolerant yamame in normal condition after heat stress. For that, a group of yamame was developed through selective breeding to have high temperature tolerance. Next, we performed a higher-temperature-tolerant test and divided into HT (for the high-temperature-tolerant) group and NT (for the non-high-temperature-tolerant) group. A week later, RNAs were extracted from the gill tissues and analyzed by examining the mRNA expression profiles using Illumina HiSeq 4000 Sequencing System. A total of 2,893 differentially expressed genes (DEGs) from the gill were identified by comparing the HT and the NT groups, then functional analysis were performed to identify associated gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Several differential biological pathways were detected and we found that the HT showed higher associated gene expression in ECM-receptor interaction, in cell adhesion molecules (CAMs), in cell junction and in adhesion pathway comparing to the gill tissue in NT. Those genes are related to the reparation of the damaged tissues and to the generation of the cytoskeleton of individuals. On this basic, we concluded that the HT may adapt quicker than the NT in normal condition after undergoing the heat stress. These findings can be used to develop high-temperature-tolerant yamame and other Salmonidae.

Biography:

Jenn-Tzong Chen is a PhD student of Institute of Polymer Science and Engineering, and Chemical Engineering of National Taiwan University. He is also an associate researcher of Isotope Application Department of Institute of Nuclear Energy Research. After graduated from the Chemical Engineering Department of Ming-Chi Institute of Technology, he studied Chemical Engineering in National Taiwan Institute Technology for one year. Then he graduated from Chemical Engineering Department of National Taiwan University with master degree. He has been trained in Forschungszentrum Jülich, Germany and Washington University in St. Louis, US and so on. At present he works for cyclotron produced radioisotope process development and application. He has published some papers and owned some patterns.

Abstract:

Lipophilicity is one of the major brain radiopharmaceutical design criteria. Alzheimer disease PET imaging agents based on lipophilicity modification are [18F]RO6958948 [1] and [18F]Florbetapir, design by replacing with a Nitrogen element either in the aromatic ring of [18F]Flortaucipir or [18F]Florbetaben. The structure of [18F]FEONM (Figure 1) is designed to provide higher lipophilicity than [18F]FDDNP. Structure modification on a certain bioactive molecule in order to increase its lipophilicity will be also possibly increasing the percentage of penetrating blood brain barrier. Increasing the blood brain barrier crossing ratio, the specificity of this active biomolecule targeting effect might be decreased. Therefore, we design an ethyl oxide modified naphthol based Alzheimer disease positron emission tomography imaging agent [18F]FEONM, in order to compare the uptake effect of Tau tangle and Beta amyloid.

PET radiopharmaceiticals for brain imaging are based on very short half-life radionuclides, most of them will be decayed in one day. One of the longest half-life organic radionuclides is fluorine-18, therefore critical step to producing PET radiopharmaceuticals online is radiofluorination reaction. The highest radiofluorination reaction yield can be made from carboxy glass reactor [2]. In carboxy glass reactor, the function of gap area (FG) [3] curve of radiofluorination yield can be approached with Gauss distribution, Gauss or Welch apodization function. After determine the radiofluorination rate constant, the length of microfluidic plug flow reactor can be designed with an analytical form based on Welch apodization function [4].

Brain hippocampus imaging relative specific binding ratio of [18F]FEONM on a Tau tangle P301S/PS19 transgenic mouse model is two time higher than cerebellum [3], Beta amyloid Tg2576 transgenic mouse model is less than two. On a triple transgenic 3xTg mouse model with both Tau tangle and Beta amyloid formed, the uptake ratio of hippocampus is fifty percent higher than cerebellum. Therefore, [18F]FEONM is a new Alzheimer PET imaging agent. Besides, other than transgenic mouse model, streptozotocin induced Tau tangle mouse model [5] also shows higher brain hippocampus [18F]FEONM uptake than control mouse.

From the transgenic mouse model imaging study, we found [18F]FEONM will uptake on both Tau tangle and Beta amyloid transgenic mouse. In comparison to [18F]FDDNP, it shows no Beta amyloid transgenic mice uptake in brain hippocampus [6]. This result represents part of the specific binding of Tau tangle transgenic mouse of [18F]FDDNP has shift to Beta amyloid. Therefore, Tau tangle and Beta amyloid uptake status can be done by [18F]FEONM in the same time for diagnosis Alzheimer disease. Radiation exposure will be half dosage compared to taking both imaging. These findings based on a new design conclude that a new PET radiopharmaceutical design has the same concept like a new radiofluorination microfluidic reactor design. Either a new chemical structure or a new mathematical model contributes an achievement.

Keywords: Alzheimer’s disease, Welch apodization gap area function, [18F]FEONM

Reference

1. Honer M, Gobbi L, Knust H, Kuwabara H, Muri D, Koerner M, Valentine H, Dannals RF, Wong DF, Borroni E (2018) Preclinical Evaluation of ¹⁸F-RO6958948, ¹¹C-RO6931643, and ¹¹C-RO6924963 as Novel PET Radiotracers for Imaging Tau Aggregates in Alzheimer Disease. J Nucl Med 59(4):675-681.

2. Hamaker K, Blessing G, Nebeling B (1990) Computer aided synthesis (CAS) of no-carrier-added 2-[18F]fluoro-2-deoxy-D-glucose: an efficient automated system for the aminopolyether-supported nucleophilic fluorination. Appl Radiat Isot 41:49-55.

3. Hsu JP, Chen JT, Lin SH, Chu KY, Tu YH, Chang KW, Huang LY, Huang YC, Farn SS, Hsu SH, Lin JJ, Lin WJ (2016) Radiofluorination process development and tau protein imaging studies of [F-18]FEONM. J Taiwan Inst Chem Eng 68:119-129.

4. Hsu JP, Chen JT, Lin JJ, Li MH, Chang KW, Chen DC, Duh TS, Lo PL, Farn SS, Luo TY, Lin WJ, Shiue CY (2019) Design of PET radiopharmaceuticals for brain imaging. Biomed J Sci & Tech Res 14(2):1-3. Doi: 10.26717.bjstr.2019.14.002527

5. Pradip KK (2015) Streptozotocin induced Alzheimer's disease like changes and the underlying neural degeneration and regeneration mechanism. Neural Regen Res 10(7):1050-1052.

6. Kuntner C, Kesner AL, Bauer M, Kremslehner R, Wanek T, Mandler M, Karch R, Stanek J, Wolf T, Müller M, Langer O (2009) Limitations of small animal PET imaging with [¹⁸F]FDDNP and FDG for quantitative studies in a transgenic mouse model of Alzheimer’s disease. Mol Imaging Biol 11:236-240.