Day 3 :
- Track 7: Applied Pharmacology
Track 8: Biotoxins
Track 9: Pharmacovigilance & Toxicologists Meeting
Location: DoubleTree by Hilton Philadelphia Airport
Chair
Shailesh P Banerjee
City University of New York, USA
Co-Chair
Marketa Bebarova
Masaryk University, Czech Republic
Session Introduction
Shailesh P Banerjee
City University of New York, USA
Title: Modes of neuropharmacological actions of taurine and its potential therapeutic usage
Time : 09:00-09:20
Biography:
Shailesh P Banerjee received his PhD in Pharmacology from University of Toronto and a MPH degree from the Columbia University. He has served as an Associate Editor of the Journal of Neuroscience 1981-84. He has been in in the Medical School Faculty in either University of Rochester School of Medicine or CUNY Medical School for last forty years or more and published more than 100 scientific research papers in reputed journals.
Abstract:
Taurine is an inhibitory neuromodulatory endogenous amino-acid in the CNS and activates GABA- and glycine-insensitive chloride channel and inhibits the glutamate NMDA receptor. We investigated taurine’s interaction with the NMDA receptor using electrophysiological, receptor binding studies and explored its long-term effects by using western blot determination of NMDA and AMPA receptor subunits expression in the rat frontal cortex. Taurine was found to modulate NMDA receptor function without affecting AMPA receptor mediated activity perhaps by partially blocking spermine-potentiated NMDA receptor activation. Furthermore, our studies revealed that inhibition of evoked responses by taurine overlapped with that by Ro-25-6981, a selective antagonist for the GLuN1/GLuN2B NMDA receptor subtype, suggesting that taurine modulates NMDA receptor by acting on the NMDA GLuN1/GLuN2B receptor subunit (s). This observation was confirmed by conducting receptor binding studies using tritiated- spermidine or-taurine that showed that taurine and polyamines may interact at a common binding site. Chronic administration of taurine caused significantincrease in the expression of NMDA GLuN2B, but not GLuN1 subunit and a significant decrease in the expression of the AMPA GLuR2 subunit. The up-regulation of the GLuN2B subunit suggests a consequence of its possible long-term interaction with taurine, and the down-regulation of the AMPA GLuR2 subunit is possibly correlated to an increased recruitment of the GLuR2-subunit-lacking, calcium-permeable subtype of AMPA receptor. Other studies in our laboratory showed that taurine is an effective anti-cataleptic and neuro-protective agent. Also, micro dialysis investigation of effects chronic administration of psychotropic drug, cocaine revealed an increase in extracellular release of endogenous taurine which may protect against deleterious effects of the substances of abuse. In addition, taurine administration was found to prevent cocaine-induced addiction by suppressing spontaneous locomotor activity and conditioned place preference. Thus taurine is a unique psychopharmacological compound with potential for a variety of therapeutic uses including as a neuro-protective, anti-cataleptic, and anti-addicting agent.
Marketa Bebarova
Masaryk University, Czech Republic
Title: Alcohol intoxication, arrhythmias and inward rectifiers
Time : 09:20-09:40
Biography:
Marketa Bebarova has completed her MD studies at the age of 25 years and her PhD studies at the age of 29 years, both at Masaryk University (Czech Republic). Subsequently, she passed a Postdoctoral research stay at Cardiovascular Research Institute Maastricht, Maastricht University (Netherlands). Nowadays, she serves as an Associate Professor and as the Head of Laboratory of Cellular Electrophysiology at Department of Physiology, Faculty of Medicine, Masaryk University. She has published 20 papers in reputed journals. She is a nucleus member of the European Working Group (WG) on Cardiac Cellular Electrophysiology (WG of the European Society of Cardiology).
Abstract:
Alcohol intoxication may induce electrocardiographic changes and arrhythmias, most frequently the atrial fibrillation (AF). In the pathogenesis of arrhythmias including AF, modifications and/or heterogeneity of inward rectifier potassium currents, namely IK1 and acetylcholine-sensitive current IK(Ach), are known to be involved. Recently, we have reported that ethanol at clinically relevant concentrations alters the rat ventricular IK1 in a dual way. We have also observed a significant inhibitory effect of acetaldehyde, the primary metabolite of ethanol, on this current. Considering missing data, we aimed to analyze ethanol-induced changes of inward rectifiers in the cardiac atria. Experiments were performed by the whole-cell patch clamp technique at room temperature in enzymatically isolated rat atrial myocytes. In contrast to the ventricular IK1, ethanol in concentrations of 20 and 80 mM (ï¾0.09 and 0.4%) showed only minor average changes of the atrial IK1. In the case of IK(Ach), both components of this current, i.e. the constitutively active and the acetylcholine-induced IK(Ach), were affected by ethanol. The constitutively active IK(Ach) was significantly increased at all examined concentrations between 2 and 80 mM (e.g. at 20 mM by 114.6±28.3%). The acetylcholine-induced IK(Ach) was affected by 8 to 80 mM ethanol in a dual way with its prevailing activation (e.g. by 29.2±17.3% at 20 mM). To conclude, the effect of ethanol on the atrial IK1 was minor compared to its effect on the ventricular IK1. Changes of the atrial electrophysiology under alcohol intoxication are likely caused by other factors, the clinically relevant action of ethanol on IK(Ach) may contribute.
Peizhen Janet Song
QNS Consulting, USA
Title: Effects of bile acid sequestration on bile acid profile and bile acid signaling in mice
Time : 09:40-10:00
Biography:
Peizhen Janet Song received her PhD from the University of Kansas, School of Medicine and her Medical Doctor degree from Shanxi Medical University. As a principal in QNS consulting, LLT, She has contributed to the success of numerous projects for her clients which include some global giants in the pharmaceutical world. Her expertise lies in R&D and regulatory affairs in life science related fields.
Abstract:
Recently, the discovery of bile acid sequestrant (BAS) as a novel anti-diabetes medication places renewed emphasis on the need for more research on the mechanism by which BASs play their pharmacological roles. BASs are non-absorbable resins and function by contraction of the bile acid (BA) pool in the body. Herein, we hypothesize that BASs exert their pharmacological function by altering serum BA composition and BA signaling in the body. Mice were fed 2% Cholestyramine (resin) in their diets for one week. The individual BA concentration and mRNAs of genes involved in BA signaling were quantified. As expected, the resin reduced total BA concentration in the liver 80%. But surprisingly, the resin increased the concentrations of CA and T-CA in the serum, causing a slightly increased total BA concentration in the serum. The mRNA of the BA-biosynthesis enzymes, Cyp7a1 and Cyp8b1, increased 180 and 100% respectively after resin feeding. However, the FXR-target gene SHP in the liver was not decreased. In contrast, the ileum Fgf15 mRNA expression was significantly decreased. Feeding the resin did not alter the BA uptake transporters on the sinusoidal membrane nor did it altered the efflux transporters on the canalicular membrane, whereas, the efflux transporters on the sinusoidal membrane of hepatocytes, Mrp 3 and 4, were increased by the resin 22 and 150% respectively. In ileum, feeding the resin increased the uptake transporter, Asbt, 66%. Collectively, the data suggest that increased expression of Cyp7a1 and Cyp8b1 in mouse liver by resin administered through the diet appears to be due to a decrease in the ileum Fgf15 signaling, rather than a decrease in hepatic FXR signaling. In addition, the increased concentration of CA in the serum may play a role in the hypoglycemic effects of the resin.
Balszuweit F
Bundeswehr Institute of Pharmacology and Toxicology, Germany
Title: Silibinin as a potential therapeutic for sulfur mustard injuries
Biography:
Balszuweit F is a pharmacist and obtained his PhD at the Free University Berlin in 2005. Along with his research activities within the Bundeswehr Medical Service, he has been concerned with research management, regulatory affairs and scientific cooperation. His research interests are focused on cell co-cultures to identify novel treatment strategies, in particular against sulfur mustard injuries.
Abstract:
Sulfur mustard (SM) is a vesicating chemical warfare agent causing skin blistering, ulceration, impaired wound healing, prolonged hospitalization and permanent lesions. Silibinin, the lead compound from Silybum marianum, has been discussed as a potential antidote to SM poisoning, but previous investigations had been limited to nitrogen mustards. Water solubility of silibinin is poor, thus a water-soluble prodrug, e.g. silibinin-bis-succinat (silibinin-BS, SIL-BS) should be desirable for rapid bioavailability as an antidote. HaCaT cells were exposed to SM (30, 100, and 300 μM) for 30min and treated thereafter with SIL-BS (10, 50, and 100μM) for 24h. Necrosis, apoptosis and production interleukin-6 and -8 were determined. SIL-BS dose-dependently reduced SM cytotoxicity, even after 300 μM exposure. Doses of 50-100 μM SIL-BS were required for significant protection. Apoptosis and interleukin production remained largely unchanged by 10-50 μM SIL-BS but increased slightly after 100 μM treatment, in particular when cells had previously been exposed to 300 µM SM. HaCaT cells, incubated with SIL-BS were lysed and investigated by LC-ESI MS/MS. Findings suggest that SIL-BS is absorbed by HaCaT cells and biotransformed into free silibinin. In summary, silibinin-BS is a promising compound for the treatment of SM injuries: biotransformation to free silibinin is possible and standard doses for clinical use (50-100 µM) provided a significant reduction of necrosis. At doses of 50 µM SIL-BS, no pro-inflammatory or pro-apoptotic effects occurred, but even pro-apoptotic effects of 100 µM SIL-BS were observed only after 300 µM SM exposure and might even be useful to eliminate cells with irreversible SM-induced damage.
Guilhem Bousquet
University Paris-Diderot, France
Title: Can cancer brain metastases be treated using humanized monoclonal antibodies? A pilot pharmacological study
Time : 10:00-10:20
Biography:
Guilhem Bousquet is a physician involved in medical oncology since 2005. After his clinical training as a fellowship in Clinical Oncology, he did his PhD thesis in Saint-Louis Hospital in Paris, where he gained an expertise in pathology, xenograft models and pre-clinical pharmacology. He performed a Post-doctoral stay in Pasteur Institute in Lille, and is now Associate Professor in Saint-Louis Hospital and member of the research unit UMR_S1165 University Paris 7/Inserm. His research team is involved in experimental pharmacology and translational research using xenograft models of human fresh tumors. He has published 41 papers.
Abstract:
Therapeutic monoclonal antibodies have widely contributed to improving survival in metastatic cancers. However, patients are at increased risk of developing central nervous system metastases, and the survival of patients with brain metastases remains poor, challenging daily practice in medical oncology. The mechanisms limiting the use of therapeutic monoclonal antibodies for the treatment of brain tumors are their inadequate transport from blood to brain through the blood-brain barrier. We performed a sequential pharmacological pilot study in a young, 36-year-old woman with metastatic breast cancer who developed HER2-overexpressing brain metastases resistant to standard treatment. With her informed consent, lumbar and ventricular reservoirs were implanted for repeated intra-thecal injections of trastuzumab, an anti-HER2 monoclonal antibody. Close monitoring of trastuzumab trough concentrations in the cerebrospinal fluid over 8 months enabled us to determine the intra-thecal drug administration schedule required to stabilize the brain metastases on magnetic resonance imaging. Because of a high clearance from the cerebrospinal fluid, and this result was unexpected, repeated injections of trastuzumab 3 times a week were required. This innovative scheme, guided by individual pharmacology and mimicking a continuous perfusion, can be used for other patients. Apart from this therapeutic effect, the rapid efflux of trastuzumab has not hitherto been described. FcRn receptors, expressed by the capillary endothelium of the blood-brain barrier, may be responsible for this efflux mechanism. Engineering of a F(ab’)2 fragment of humanized monoclonal antibody against HER2 could prevent the binding of immunoglobulin to FcRn, and the mechanism of efflux.
Aditya Arya
University of Malaya, Malaysia
Title: Rosmarinic acid mediates glucose transporter protein (GLUT-4) by upregulating C-peptide and insulin in diabetic rats
Time : 10:20-10:40
Biography:
Aditya Arya is a Senior Lecturer of Pharmacology at Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia. He has received PhD in Pharmacology from the University of Malaya and have published over 30 peer-reviewed papers in reputed Journals. He has been serving as a reviewer for many ISI Journals including editorial board member. His research emphasizes on cellular and molecular Pharmacology, specifically to evaluate the co-relationship between inflammation, cancer and diabetes. He is expert in designing, development and mechanistic prediction of drug molecules on the living system and their role on G-protein-coupled receptors in islet function and stimulus-response coupling in mouse and human islet β-cells. Moreover, to detect and identify novel insulin secretagogues.
Abstract:
The study aimed to investigate the potential role of Rosmarinic acid (RA) in the management of diabetes. RA is an ester of caffeic acid found in various plants which have reflected beneficial effects on different disorders. In our study design, the normal rats were treated with different doses of RA to evaluate acute toxicity. Further on the basis of acute toxicity results, two doses of RA were selected for antidiabetic study on STZ-nicotinamide induced diabetic rats. Diabetic rats were fed 50 and 100 mg/kg body weight of RA daily for 90 days and weekly measured with certain biochemical parameters, blood glucose, insulin, C-peptide, lipid profiles and body weight. At the end of the study period, all the group animals were sacrificed and the blood glucose, insulin, C-peptide, and HbA1c levels were determined in the serum of rats. In addition, histology of pancreas and the expression of glucose transporters (GLUT 1, 2 and 4) proteins were assessed in skeletal muscles and pancreatic tissues. The overall study result demonstrated that RA has potentially normalized the elevated blood glucose levels and significantly improved serum insulin, C-peptide and HbA1c levels thereby reducing cholesterol and bad lipids. Furthermore, the histology result reported recovery in the structural degeneration in the pancreatic tissues, and the western blot analysis showed the significant increase in the GLUT-2 and GLUT-4 protein expression in the skeletal and pancreatic tissues. Altogether, we may postulate that the translocation of glucose transporter proteins (GLUT-4) accelerates the insulin-mediated glucose uptake in muscle and adipose tissue. Thus, we may conclude that RA could be the potential targets for the management of diabetic complications and needs further in depth mechanistic studies at molecular level.
Smita Shenoy
Manipal University, India
Title: A retrospective analysis of serious adverse drug reactions in a tertiary care hospital
Biography:
Smita Shenoy is an Additional Professor of Pharmacology at Kasturba Medical College, Manipal, India. She has a total of 13 years of teaching experience. She has more than 30 publications in national and international journals.
Abstract:
Serious adverse drug reactions (ADRs) are associated with morbidity and mortality and result in financial burden to the patient and society. A retrospective observational study of serious adverse drug reactions over 1 year in a tertiary care hospital was carried out. Risk factors for serious ADRs were identified (Chi square test). Average direct cost of managing the adverse drug reaction was calculated. Of the 985 adverse drug reactions reported, 95 were serious ADRs. About 49.47% (n= 47) of patients were males and 50.53% (n=48) were females. The mean age of patients with serious ADRs was 48.63±15.59 years. Of the total hospital admissions, 0.05% was due to serious ADRs. The median duration of hospital stay due to serious adverse drug reaction was 7(4, 11) days. Hematological disorders (41.05%) were the most common serious ADRs followed by electrolyte disturbances (18.94%) and hypersensitivity reactions (13.68%).The common serious ADRs were leucopenia (15.78%), febrile neutropenia (12.63%) and hyponatraemia (11.57%). Anticancer drugs constituted the majority of the suspect drugs. Concomitant medications included anticancer drugs, antitubercular drugs, antiretrovirals, antileprotic drugs, antimicrobials, steroids and analgesics. The risk factor for developing serious ADRs was gender (females, p<0.04 versus males), coexisting disease (p< 0.001) and more than two concomitant medications (p< 0.01). The average total direct cost was Rs. 7306.91 per serious ADR. Since ADRs are associated with a wide variety of drugs and can affect any organ in the body, it is important to detect them early and have appropriate preventive strategies.
Bharti Chogtu
Manipal University, India
Title: Analysis of drug induced maculopapular rashes in a tertiary care hospital
Biography:
Bharti Chogtu has done MD in Pharmacology. She is Associate Professor in department of Pharmacology. She has published about 45 papers in reputed journals.
Abstract:
Adverse cutaneous reactions to drugs affect about 2-3% of all hospitalized patients. Maculopapular rash is one of the most frequent cutaneous clinical manifestation of non-immediate allergic reactions due to drugs. This study was done to analyze drug induced maculopapular rashes in patients in a tertiary care hospital. It was a prospective study done in patients having maculopapular rashes secondary to drugs for a period of six months. The data was recorded which included demography of the patients, drugs which were implicated in these reactions. The cutaneous adverse drug reactions were then assessed for determining causality and severity. Descriptive statistics was used to analyze the data. A total of 104 patients of drug induced maculopapular rashes were included. The age of the patients ranged from 2-73 years with a median range of 39 years (IQ 21.25-51.75). Males constituted 52.88% of total patients. The most common group of drugs causing maculopapular rash was antimicrobial agents (42.3%) followed by antiepileptics (20%) and non-steroidal anti-inflammatory drugs (14.4%). Of antimicrobial agents, amoxicillin was the most common causative agent followed by antitubercular drugs and nevirapine. Of all the individual drugs, phenytoin was implicated in maximum number (14.4%) of drug reactions. As per WHO causality assessment, 75% of the adverse reactions were possible, 24% probable and 1% certain. The severity of adverse drug reactions was moderate in 85.5% and the remaining reactions were categorized as mild. The offending drug was stopped in 84.6% of patients. 89.4% patients had recovered whereas adverse event continued in the remaining at the time of reporting. Thus, there is a need to monitor cutaneous adverse drug reactions by physicians prescribing antimicrobials and antiepileptics.
Biography:
Shalini Adiga has completed her MD Pharmacologyin the year 2000 from Kasturba Medical College, Manipal, Manipal University. She is the Professor and Head of Pharmacology. She has 27 publications in various journals.
Abstract:
Steven Johnson Syndrome (SJS) is a life threatening muco-cutaneous adverse reaction to medications with high mortality. The aim of this study was to retrospectively review the relationship between drugs induced SJS, the causative agents and the patient characteristics. In patients reported with SJS over a period of 2yrs at a tertiary hospital, data were collected for demographic information, causative drugs, time interval between drug intake and onset of symptoms, duration of hospital stayand clinical outcome. Causality assessment was carried out using the WHO probability scale and data was analysed using SPSS software (version 20).The study included 47 cases of which 29 comprised females. Of the total cases 10 patients were less than 20yrs and 4 patients more than 60 yrs of age with a median age of 40 yrs. Antimicrobials (32%), anti-epileptics (32%) and non-steroidal anti-inflammatory drugs (17%) were the most commonly associated drugs. Amongst antimicrobials fluroquinolones were the most frequent causative drugs followed by cephalosporins and antiretroviral agents. Phenytoin was the commonly implicated antiepileptic drug.The onset of reaction was seen to be most rapid with antibiotics and NSAIDs, with median onset to reaction time being 3 and 4.5 days respectively. Anti-epileptics had the longest latency of 21 days to onset of reaction. Patients developing SJS following antibiotic use had the longest duration of hospital stay, while patients who had no history of drug intake had the least duration (median of 12 days v/s 2.5 days) according to the WHO causality assessment all cases had ‘probable’ association with prior drug intake.
Meena Kumari Kamal Kishore
Manipal University, India
Title: Analysis of hematological adverse drug reactions due to anticancer drugs in a tertiary care hospital
Biography:
Meena Kumari Kamal Kishore has completed her MD in pharmacology from Manipal Academy of Higher Education, currently known as Manipal University. She is an Associate Professor in Pharmacology. She has published more than 30 papers in reputed journals. She is also a moderator for reporting the adverse drug reactions for the pharmacovigilance section of their department. She has also published articles under pharmacovigilance.
Abstract:
Aim: To analyze the hematological adverse drug reactions (ADRs) due to anticancer drugs in a tertiary care hospital. Material & Methods: It was a prospective, observational study done on patients admitted to the medical oncology ward of Kasturba Hospital, Manipal from October 2014 to March 2015. All patients developing hematological adverse drug reactions were included. CDSCO proforma was used for reporting ADRs. Causality was assessed using Naranjo Algorithm. Severity was assessed using Hartwig and Siegel severity scale. Preventability was assessed using Modified Shumock and Thornton preventability scale. Results: Forty patients in this study were found to have hematological ADRs, of which twenty one were males and rest females. Highest incidence was seen in 21-40 y (50%) age group compared to least in 1-20 y (7.5 %) age groups. The most common ADRs were anemia (27.5%), followed by thrombocytopenia (25%), neutropenia (20%), leucopenia (17.5%) and bone marrow suppression (10%). Antimetabolites group contributed to majority of ADRs (30 %), followed by anticancer antibiotics (25%), alkylating agents (15%), topoisomerase-II inhibitors (12.5%), platinum compounds (10 %), microtubule inhibitors (5%) and miscellaneous (2.5%) comprising the least. Totally 24 patients recovered from the ADRs. Majority (97.5 %) of the ADRs were of moderate severity. The results of causality assessment of most of the cases were possible (85 %). All the ADRs were not preventable. Conclusion: Anticancer drugs are known for multiple adverse effects. Early detection and timely management of the ADR with appropriate therapy would reduce the fatal ADRs.
- Young Researchers Forum
Location: DoubleTree by Hilton Philadelphia Airport
Session Introduction
Nilmara de Oliveira Alves
University of São Paulo, Brazil
Title: The effects of biomass burning from Amazon region in human lung cells
Time : 11:00-11:10
Biography:
Nilmara de Oliveira Alves is a post-doctoral fellow in the School of Medicine of the University of São Paulo. The researcher, who recently received her doctoral title, focuses on atmospheric pollutants and especially on their effects on health using both in vitro and in vivo tests. For that end, she conducts interdisciplinary projects comprisinggenotoxicity, DNA repair, pathology and atmospheric chemistry.
Abstract:
The Brazilian Amazon population has been negatively affected by biomass burning. The majority of forest fire hotspots in the Amazon take place in the deforestation arcwith a population of over 10 million inhabitants.However, there are few studies to understand the mechanism of action of aerosols in human health. Thus, we collected filters with particulate matter (PM10) to investigate the effects of biomass burning at molecular and cellular levelsusing human lung cells (A549). The chromatography-mass spectrometry analysis showed the presence of carcinogenic and mutagenic compounds. After chemical analysis, we defined a dose below the limit established by the World Health Organization (30 µg/m3). After 24 hours of exposure,there was an increase of pro-inflammatory cytokines and in ROS generation, in a dose and time dependent manner. Besides, there was an induction of cell cycle arrest at G1 phase, as well as an increase in the expression p53 protein and formation of DNA strand breaks. After 72 hours, we detected a significant increase of cells in the sub-G1 fraction, indicating apoptosis. Additionally, we observed the phosphorylation of H2AX (γ-H2AX), which correlated with the activation of caspase 3, suggesting that the induction of γ-H2AXmay be associated with the DNA fragmentation during apoptosis. We also observed that necrosis is a cell demise pathway induced by PM10. This study shows an important advance in understanding the toxic cellular and molecular effects induced by PM10 that can be related to the increase potential of human health impacts in the Amazon region.
Ekwere M R
University of Calabar, Nigeria
Title: Haematological indices of male rats following oral exposure to a mixture of cypermethrin and dimethoate (uppercott insecticide)
Time : 11:10-11:20
Biography:
Ekwere M R is a senior lecturer and a PhD student of Uboh F E. Ekeruke M B is a student under the supervision of Uboh, assisted by Ekwere
Abstract:
The present study evaluated the heamatological parameters of male rats exposed to oral administration of daily doses of 10 mg/ kg and 20 mg/kg uppercott (a mixture of cypermethrin and dimethoate belonging to organophosphate and pyrethroid groups). They were sacrificed after 28 days. Results obtained showed a dose dependent decrease in red blood cell (RBC), packed cell volume (PCV), Mean corpuscular haemoglobin (MCH) and corpuscular heamoglobin concentration (MCHC). White blood cells (WBC), lymphocytes and neutrophils increased significantly (p<0.05) while eosinophiles had no significant change (p>0.05) in concentration comparison with control. Thus, uppercott may induce haematoxicity in mammals.
Somnath Paul
CSIR, India
Title: Epigenetic susceptibility of long interspersed nuclear elements-1 towards arsenic induced DNA damage: A predictive biomarker
Time : 11:20-11:30
Biography:
Somnath Paul has completed his PhD dissertation work in 4.5 years, working as graduate student at CSIR-Indian Institute of Chemical Biology since 2010. He is in the process of writing his thesis, to be submitted to Jadavpur University. He has done his Bachelors in Physiology, from Presidency College, Kolkata and Masters in Genetics from University of Calcutta. He has published 7 papers in peer reviewed journals. His major thrust area of research is epigenetics and have future interest in doing research on epigenetic alterations in human diseases and understand the role of epigenetic mechanisms in various patho-physiological outcomes.
Abstract:
Arsenic exposure mainly through drinking water has increased over the years, presently affecting more than 137 million individuals worldwide. One of the prime facets of arsenic toxicity is disruption of epigenetic profile within the cell, leading to various outcomes like silencing or activating genes from their normal state of expression. In the field of toxicology, a recent emphasis lies with the probability of epigenetic alteration and it’s indulgence in DNA damage. Long Interspersed Nuclear Elements (LINE-1) have been studied as a prospective candidate to understand the global genomic methylation profile. Activity and methylation profile of these transposable elements have been associated with various forms of cancers as well as genomic stability. Since arsenic exposure yields a high degree of DNA damage, in our present study we are focusing on the epigenetic risk factor associated with DNA damage and L1-methylation. A total of 100 arsenic exposed samples were recruited from the district of Murshidabad, West Bengal. Cytogenetic damage was analyzed using micronucleus (MN) assay from lymphocytes. When the cohort comparison between arsenic exposed individuals distributed among low, medium and high degree of cytogenetic damage based on the MN frequency, we observed that there was a positive association (p<0.01) between the lowering of methylation profile of LINE-1 with increase in MN frequency. Our data suggests a positive influence of LINE-1 methylation may be related to alteration in chromatin landscape, leading to subsequent genomic instability upon arsenic mediated oxidative DNA damage. LINE-1 methylation hence can be predictive biomarker to epimutagenic events by arsenic.
Baragamaarachchi R Y
University of Colombo/ Industrial Technology Institute, Sri Lanka
Title: Recommendations to enhance reproducibility and reliability in comet assay
Time : 11:30-11:40
Biography:
Baragamaarachchi R Y is a first year PhD student who graduated with a first class in BSc Genetics from University of Bangalore. She obtained MSc in Molecular Life Sciences in 2014 from University of Colombo, Sri Lanka. Her research interests lie in Molecular Biology, Immunology, Medicinal plants, Genetics and Microbiology. She has served as a resource person at workshops based on Immunological techniques and awarded at 2nd International Conference on Frontiers in Molecular Life Sciences held in Sri Lanka (2014), for outstanding poster presentation.
Abstract:
Comet assay is a very sophisticated technique use to assess plethora of DNA damages at the level of individual cells and can be applied in a range of fields from human and environmental biomonitoring, routine genotoxicity assessment, DNA repair studies, and clinical studies to molecular epidemiology. Due to its demonstrated advantages and applications it is increasingly acceptable by regulatory authorities as a part of battery of assays used for regulatory submissions in genetic toxicology. Yet the major drawback of this technique is the unreliability in making reproducible data, due to miscellaneous conditions used in different laboratories and due to lack of understanding of the critical steps. Comet assay was performed according to the protocol described by Raymond Tice, with some modifications. Some critical steps that affect the final outcome of the assay were optimized during thisstudy. As the concentration of low melting agarose greatly influenced the comet formation and assay reproducibility, melting of agarose in a boiling water bath was found to be better than melting using a microwave oven. Optimal agarose solidification time was 30 min at 4°C to attain adequately solidified agarose layers which reduce the fragility. Optimal lysis time was 2 h at 4°C. Alkaline unwinding time was increased up to 30 min for adequate denaturation of DNA. The best way to calculate the required voltage to get a voltage drop of 1 V/cm, was to use the spreadsheet developed by Gunnar Brunborg. Electrophoresis for 45 min was found to be sufficient to obtain a considerable DNA migration. However, a small factor, that one would not consider, significantly matters the final outcome and the assay reproducibility.
Deniz Özkan Vardar
Hitit University, Turkey
Title: Cytotoxicity of silver sulfide quantum dots evaluated by the MTT cell culture assay in Hela Cells
Time : 11:40-11:50
Biography:
Deniz Özkan Vardar has completed his bachelor’s and master degree at Gazi University. She is PhD student in Department of Nanotechnology and Nanomedicine, Hacettepe University. Her academic work is focused on nanotoxicology especially nano genotoxicology. She has published 2 papers about genotoxicity. She has worked for more than 5 years in the Departments of Health Programs, Hitit University.
Abstract:
Nanomaterials have received enormous attention for their potential applications in biology and medicine. A key issue in evaluating the utility of these materials is the assessment of their potential toxicity−either due to their inherent chemical composition or as a consequence of their nanoscale properties. Quantum dots are an example of a nanomaterial that has been shown to be useful as an alternative to fluorescent dyes for use in biological imaging, due to their bright fluorescence, narrow emission, broad UV excitation, and high photo stability. In addition to labeling of cellular structures in vitro, several groups have demonstrated the use of quantum dots (QDs) for fluorescence imaging in vivo. Several in vitro and in vivo studies have been cited in the literature as demonstrating the lack of evidence for QD-induced cytotoxicity. The sensitivity of the cytotoxicity assay used differs depending on the different mechanisms, which lead to cell death. The MTT assay is simple and rapid to use, which determines, the metabolic activity of the mitochondria can be determined. We aimed to evaluate the cytotoxic effects of silver sulfide quantum dots coated with 2-Mercaptopropionic acid (2MPA) and Meso-2, 3-dimercapto succinic acid (DMSA) because of the lack of studies in this area. Cytotoxicity was evaluated by the MTT assay in HeLa cells. Our results showed that Ag2S QDs were not cytotoxic effects after 24 h exposure.
Adetutu Adewale
Ladoke Akintola University of Technology, Nigeria
Title: Anti-malaria activities of selected plants and gas chromatography-mass spectrometer chemical profiling of aqueous bark extract of Prosopis africana (Guill &Perr)
Time : 11:50-12:00
Biography:
Adetutu Adewale obtained his Ph.D from Department of Biochemistry, Ladoke Akintola University of Technology, Ogbomoso, Nigeria
Abstract:
Background: The development of resistance to currently known conventional anti-malaria drugs has necessitated search into more potent and less toxic anti-malaria drugs of plant origin. Objective: Hence, this study aimed to document plants commonly used to treat malaria in Ilorin metropolis, Nigeria and validate the traditional claims using in vivo anti-plasmodial tests. Methods: Semi-structured questionnaires (70) were used to explore the ethno-botanical practices amongst the traditional healers. The most common species cited were identified, authenticated and their aqueous extracts were screened for antimalarial activities using Plasmodium berghei (NK 65 chloroquine sensitive) and chloroquine as the malarial parasite and positive control respectively. For in vivo anti plasmodial testing, the mice were infected with 1 × 107 parasitized erythrocytes and plant extracts were subsequently administered orally for suppressive, prophylaxis and curative assays. Percentage parasitemia was estimated by standard microscopy and haematological parameters were also measured using standard analyser. Results: Seventy traditional healers from Ilorin metropolis, Nigeria were involved in the study. Forty-three species were recorded with their local names and parts used in the traditional therapeutic preparations. Ten plants with highest frequency of citation (Cymbopogon citrates (17.1%),Azadirachta indica (12.9%), Prosopis africana (12.9%), Vernonia amygdalina(11.4%), Khaya grandifoliola (10%), Terminalia glaucescens (10%), Ziniber officeinale(7.1%), Citrus paradise (7.1%), Parquetina nigrescens (7.1%), Psidium guajava (7.1%),) were selected and investigated for anti-malaria activities. The aqueous extractsof all the selected plants showed significant (p<0.05) anti-malaria activities. P. africana bark extract at 200 mg/kg body weight had the highest chemo-suppressive effect (90.02%) in comparison with other plant extracts and the standard, chloroquine (61.70%) on the 8th day. In addition, the maximum mean survival time (MST) of 23 days were observed in animals administered with P. africana and chloroquine. The extract of P. africana was further analysed for possible bioactive components using Gas Chromatography-Mass Spectrometer (GC-MS). The GC-MS analysis revealed that the aqueous bark extract of P. africana contained lipid (eight), phytochemical (sixteen) and essential oil (eighteen) components. The histological analysis of the liver revealed that the extract of P. africana was able to protect the liver against B. bergei induced damages. Conclusion: Most of the species tested had some antiplasmodial effects, which to some extent supports their traditional inclusion in herbal preparations for treatment of malaria. The bioactive components identified may be responsible for the observed antimalarial activity of P. Africana extract.
Sharma S
Guru Nanak Dev University, India
Title: Mitigating effect of cow’s urine against oxidative stress and acetylcholinestrase in rat brain and blood exposed to Chlorpyrifos
Time : 12:00-12:10
Biography:
Shelly Sharma is a research fellow pursuing doctorate from Guru Nanak Dev University, Amritsar, India. She has specialization in the field of Cytogenetics which includes genotoxicity and toxicity of pesticides. She has won Young Scientist Award in an international conference.
Abstract:
Humans are exposed to pesticides and insecticides either directly or indirectly. Exposure to these pesticides may lead to acute toxicity to mammals and non-target organisms. Chlorpyrifos (CPF) is a broad spectrum insecticide widely used in various countries of the world. The aim of the present study was to assess the toxicity associated with chlorpyrifos exposure and possible mitigating effect of cow urine against oxidative stress, activity changes in antioxidant enzymes and inhibition in activity of cholinesterase in rat brain and blood induced by chlorpyrifos. For this purpose LD50 was determined and rats were orally administered with 1/8th of LD50 (19 mg/kg b.wt). Brain and blood samples were taken after 24 hrs, 48 hrs and 72 hrs of treatment. A significant decrease in the activity of AcHE, CAT, SOD and GST was observed along with the increase in MDA levels of both brain and blood in chlorpyrifos treated groups as compared to control. Cow urine treated groups show increase in AChE while decrease in MDA level groups as compared to CPF treated group. The study indicates that cow urine has mitigating effect against toxicity induced by CPF. Thus, it can be used as an antioxidant supplement. Cow urine is considered rich in vitamin A, E and volatile fatty acids which provide antioxidant potential to it.
Rajlaxmi Basu
Institute of Hematology and Transfusion Medicine, India
Title: Health status of sewage workers: A case study
Time : 12:10-12:20
Biography:
Rajlaxmi Basu aged 27, a graduate student, carrying out her research work at Institute of Haematology and Transfusion Medicine, Medical College, Kolkata, India, for award of PhD degree from University of Calcutta.
Abstract:
Awareness of sewage workers to occupational exposure is growing very slowly in many developing countries due to which they often tend to work without proper protective clothing, resulting in a number of health problems. Lead (Pb) and cadmium (Cd) are present in sewage water and workers are exposed to these metals by unprotected handling during work. These heavy metals exposure are responsible for DNA damage and lowering of blood total iron (Fe) concentration. Zinc (Zn) is a suitable element for promoting metallothionin expression and binds the free cadmium. The total suspended solids (TSS), total dissolved solids (TDS), Pb and Cd were estimated in sewage water. The whole blood Zn and Fe concentration (estimated by energy dispersive X-ray fluorescence), Pd and Cd (estimated by GF AAS) were also estimated. Genotoxicity especially DNA damage was studied by comet assay. It was observed that there are significant differences (p<0.05) of lead and cadmium concentration in blood for exposed population i.e. sewage workers (according to through interview and observation they are non addicted) when compared with control population (non-sewage workers). The DNA damage was also observed to be significantly (p<0.001) higher in exposed groups but their blood iron concentration was significantly lower, which may be the reason for their tendency for retention of blood cadmium and make them more susceptible. Besides, they shows a highly significant depletion (p<0.001) in Selenium (Se) concentration (estimated by energy dispersive X-ray fluorescence) in their whole blood and this may be one of the cause for their lowered antioxidant level and responsible for premature ageing and different other health complication. The higher level of DNA damage (estimated by Single Cell Gel electrophoresis) is also dependent on exposure time and blood heavy metal concentration. The present study also indicates aged workers are rich in blood zinc concentration, which is a suitable indicator as this essential trace element zinc acts as an antidote for the toxic element cadmium. The present study helps to assess the health hazards of sewage workers and more susceptible groups in young compared to aged groups and as per questionnaire and estimation, the young groups have lower blood zinc level as well as different food habits, etc. that may cause DNA damage.
Nilmara de Oliveira Alves
University of São Paulo, Brazil
Title: In vivo imaging of ICAM-1 targeted nanoparticles in XPC and wild mice exposed to ambient air pollution in São Paulo, Brazil
Time : 12:20-12:30
Biography:
Nilmara de Oliveira Alves is a post-doctoral fellow in the School of Medicine of the University of São Paulo. The researcher, who recently received her doctoral title, focuses on atmospheric pollutants and especially on their effects on health using both in vitro and in vivo tests. For that end, she conducts interdisciplinary projects comprising genotoxicity, DNA repair, pathology and atmospheric chemistry.
Abstract:
Atmospheric pollution is an environmental risk factor in large urban centers. São Paulo Metropolitan Area (SPMA), located in the southeast of Brazil, has a population of nearly 20 million people and 8 million vehicles.It is long known that exposure to air pollutants can cause various health effects such as increased inflammatory response and DNA damage. One of the most versatile defense mechanisms against the accumulation of DNA damage is the nucleotide excision repair (NER), which includes the XPC protein.Different studies have shown that knockout mice in the XPC gene have an increased occurrenceof lung tumors.However, the effects of DNA damage caused by air pollutants regarding the lung inflammatory response are largely unknown. In this study, we injected intravenouslyIntercellular Adhesion Molecule-1(anti-ICAM-1) targeted nanoparticles and evaluated the response using IVIS spectrum. Mice were exposed an accumulated dose (concentration vs time of exposure in hours) of 600 µg.m-3during 1 hour, which corresponds to an average concentration of 25 µg.m-3in 24 hours.The next day after exposure, we observeda significantly stronger fluorescent marker for anti-ICAM in the polluted group than the filtered air group. Furthermore, among the animals exposed to ambient pollution, XPC mice have shown a stronger inflammatory response relative to wild mice. The expression of pro-inflammatory cytokines in the lungs is currently being evaluated.These data demonstrate that exposure to ambient air pollution in São Paulo promotes the acute inflammatory responses in mice, especially in knockout mice in the XPC gene.
Farhat Naz
All India Institute of Medical Sciences (AIIMS), India
Title: Bio-distribution and urinary excretion of Gold nanoparticles (GNPs) following systemic administration: A long term in-vivo study
Time : 12:30-12:40
Biography:
Farhat Naz has completed her PhD in 2014 from All India Institute of Medical Sciences, Delhi in cancer nanomedicine and doing Postdoctoral research in same institute. She is Research Associate in the Dept of Pathology, AIIMS. She has published 3 papers in international journals of repute.
Abstract:
Gold nanoparticles (GNPs) have shown a great potential for use as vehicle for drug delivery and other systemic use. However, long term toxicity remains a major concern. Very few long term in-vivo study pertaining to organ specific biodistribution, sequestration and excretion of GNPs are available. We investigated the long term (90 days) biodistribution, sequestration kinetics, and excretion of ultrafine gold nanoparticles after a single intravenous administration in mice. The sequestration pattern of three different sizes of GNPs 2±0.5 nm, 5±1 nm, and 10±2 nm, at a high dose of 1250 µg/Kg was determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) in various organs such as lungs, liver, spleen, heart, kidney, brain, as well as in blood and urine. GNPs of all three sizes showed highest accumulation in spleen (µg/gm of the tissues) around 15 days after injection and reached near basal level at 90 days. Low concentration of GNPs was detected in brain after 1 day without any residual of GNPs after 30 days. Ultrastructural study also showed few GNPs in brain tissue in lysosome. Renal sequestration was also low indicating reduced nephrotoxic potential. GNPs could be detected in urine till 30 days indicating near total excretion of GNPs following single injection in 1 month. No significant toxicity was documented by normal hemogram, serum biochemistry, and tissue histology. No abnormality was detected in survival, behaviour, skin and hair colour, weight and food intake. Therefore, we concluded that the ultrafine GNPs are mostly excreted out through urine without any systemic toxicity following high dose intravenous administration and it may be safe for systemic use.
Baragamaarachchi R Y
University of Colombo/ Industrial Technology Institute, Sri Lanka
Title: Cytotoxic and genotoxic potential of Walidda antidysenterica on human lymphocytes- A herb used in Sri Lankan traditional medicine
Time : 12:40-12:50
Biography:
Baragamaarachchi R Y is a first year PhD student who graduated with a first class in BSc Genetics from University of Bangalore. She obtained MSc in Molecular Life Sciences in 2014 from University of Colombo, Sri Lanka. Her research interests lie in Molecular Biology, Immunology, Medicinal plants, Genetics and Microbiology. She has served as a resource person at workshops based on Immunological techniques and awarded at 2nd International Conference on Frontiers in Molecular Life Sciences held in Sri Lanka (2014), for outstanding poster presentation.
Abstract:
Walidda antidysenterica (Apocynaceae) is widely used in traditional medicinal practices in Sri Lanka and in Asia as a remedy for respiratory disorders, hematuria, spermatorrhoea, chest affections, helminthic disorders, dressing the oozing wounds, jaundice, haemorrhoids, epilepsy, rheumatoid arthritis, osteoporosis, bacterial diseases and gut mobility disorders. Juice extracted from the bark is administrated to treat mouth sores. Flowers are used to treat snake bites, and leaves are used to treat skin disorders such as psoriasis, dermatitis etc. Despite of its medicinal value, it was reported that this plant contain pyrrolizidine alkaloids (a potent toxic compound). Therefore the current study investigated the in vitro toxicity of ethanol leaf, stem bark and flower extracts of W. antidysenterica. The cytotoxic and genotoxic activity of extracts were evaluated against human lymphocytes using trypan blue dye exclusion assay and alkaline comet assay. All extracts exhibited decrease in cell viability (< 70%) at concentrations above 50 µg/ml following 18 hour exposure, except flower extract which retained >80% cell viability even at 1000 µg/ml. Comet assay results indicated that the leaf extract induced DNA damages at the concentrations of 30, 40 and 50 µg/ml (p<0.05) and stem bark extract induced DNA damages at the concentrations of 40 and 50 µg/ml (p<0.05) compared to positive control (H2O2). Flower extract did not induce DNA damages even at 50 µg/ml (p>0.05). In conclusion, the results suggest that the flower extract was neither cytotoxic nor genotoxic whereas leaf extract showed significant cytotoxic and genotoxic activity compared to stem bark extract in a concentration dependent manner.