Day 3 :
CSIR-National Geophysical Research Institute
Keynote: Recent advances in the determination of elemental impurities in pharmaceuticals and drugs â€“ status, challenges and moving frontiers
Time : 11:00-11:30
Dr. Vysetti Balaram (DOB 1st June, 1951) received M.Sc. (1974) and Ph.D. degrees (1979) in Chemistry from the Andhra University, Visakhapatnam. Dr. Balaram is currently Emeritus Scientist (Former Head & Chief Scientist of Geochemistry Division) at the CSIR-National Geophysical Research Institute, Hyderabad. His research areas include trace element geochemistry, environmental chemistry, spectroscopy, pharmaceutical sciences and reference materials. He is an expert of ICP-mass spectroscopy (quadrupole, magnetic sector & time-of-flight) and their applications in different areas of earth, environmental and pharmaceutical sciences. He has >275 publications in international peer-reviewed journals, with ~2000 citations (h-index 23 & i10-index 52) and guided 4 PhD students, few postdoctoral and hundreds of PG students from different universities across the country. He is also recipient of several prestigious national and international awards such as "National Geoscience Award" from Government of India, New Delhi, "S. Narayanaswamy Award" from Geological Society of India, Bangalore (for his contributions in economic geology); "Eminent Mass Spectrometrists Award" from Indian Society of Mass Spectrometry, Mumbai; "Mantripragada Gold Medal” from Indian Society of Applied Geochemists, Hyderabad and Lifetime Achievement Award for Excellence in Science and Technology from ISAS-Kerala (for his contributions in mass spectrometry & analytical chemistry).
Elemental impurities have been regulated for many decades not only in environmental materials and food products, but also in pharmaceutical and drug products. Metal impurities such as As, Cd, Cu, Pb, Hg, V and Pt in pharmaceuticals and drugs may originate from several sources such as raw materials, catalysts, metal reagents and even manufacturing equipment. An account of the recent changes in the European Pharmacopoeia (EP), the United States Pharmacopoeia (USP) and the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) regulations for inorganic impurities and new strategies to be adopted for heavy metals analyses will be presented. This presentation will also describe the need and scope of metallic impurity profiling and current trends in pharmaceutical research. Rapid screening methods in quality control operations, and a brief account of the classical spectrophotometry and the role of various instrumental methods such as atomic absorption spectrometry (AAS), X-ray fluorescence spectrometry (XRF), instrumental neutron activation analysis (INAA), inductively coupled plasma atomic emission spectrometry (ICP-AES) and the inductively coupled plasma mass spectrometry (ICP-MS) for the accurate determination of inorganic impurities in pharmaceutical samples will be presented. Further developments in ICP-MS have become very significant in the quality control of bulk drugs and pharmaceutical industry. Recent advances in sample preparation, speciation analysis, quality issues, and application of laser ablation ICP-MS and high-resolution ICP-MS in pharmaceutical analysis and possible future trends will also be discussed.
- Clinical Toxicology | Reproductive Toxicology
Eva Cecilie Bonefeld-Jřrgensen
Aarhus University, Denmark
South China Agricultural University, China
Tel Aviv University, Israel
Time : 09:20-09:40
Yoram Oron has completed his PhD from the Hebrew University, Jerusalem and Post-doctoral studies from University of Virginia, School of Medicine. He is currently. Professor Emeritus of Pharmacology at the Sackler Faculty of Medicine, Tel Aviv University and the Chief Scientifi c Offi cer of two drug startup companies. He has published more than 110 papers in reputed journals and has been serving as a Reviewer in a number of reputed journals.
Extensive ischemia results in massive (≥80%) death of ganglion cells (GC) and a virtually complete loss of function as judged by b-wave disappearance in ERG. Hence this type of protocol is suited only to study the prevention of ischemic damage, but not post-insult treatment and eff ects on the extent and kinetics of recovery. We, therefore, studied the eff ects of COX species inhibition on mild ischemic damage (30% decrease in GC). Selective COX1 inhibition had no eff ect, while selective COX2 inhibitor, Vioxx, markedly improved retinal function (ERG b-wave amplitude) recovery aft er initial damage. Although GC number was also aff ected by Vioxx, the eff ect was not statistically signifi cant at this low level of damage. Vioxx also potentiated ischemic induction of HSP70. Our results strongly suggest the involvement of COX2 (possibly via inhibition of HSP70 induction) in the mechanism of retinal ischemic damage. Moreover, we propose that studies of neuro-protection at
low level damage should use functional assays, such as ERG or behavioral measurements, to follow the effi cacy of the treatment.
Aarhus University, Denmark
Title: Activation of the estrogen receptor by human serum extracts containing mixtures of perfl uorinated alkyl acids from pregnant womenâ€™s serum
Time : 09:40-10:00
Christian Bjerregaard-Olesen is defending his PhD thesis entitled, “Perfl uoroalkyl acids in serum of Danish pregnant women: Levels, time trends, extraction and ex vivo xenoestrogenicity” on October 21st 2016 in Deparment of Public Health, Aarhus University in Denmark. Additionally, he has completed his Master’s degree in Chemistry. He has published six papers in peer-reviewed journals and further four are in preparation.
Humans are exposed to a variety of perfl uorinated alkyl acids (PFAAs). Several studies have found xenoestrogenic activity of single PFAAs. Studies on mixture eff ects of the PFAAs are however sparse. In the present study, we aimed to determine the xenoestrogenic activity in human serum extracts containing mixtures of PFAAs. Recently, we developed a method to
extract the PFAAs from serum with simultaneous removal of endogenous hormones and interfering steroid metabolites. We used this method to extract the PFAAs from serum of 397 Danish pregnant women followed by analysis of estrogen receptor (ER) transactivation using MVLN cells carrying an estrogen response element luciferase reporter vector. Using 17β-estradiol
(E2) concentration-transactivation curves, we calculated the E2-equivalents (EEQ) for the extracts containing the PFAAs. 52%
of the PFAA serum extracts agonized the ER transactivation and 46% enhanced the E2-induced ER transactivation. We found positive associations between the ER transactivation and the PFAA serum levels. For the relatively few PFAA extracts that antagonized the ER in the presence of 24 pM E2 (n=38, 10%), we found inverse associations between the ER transactivation and the PFAA serum levels. Th e results indicated that the PFAA extracts induced the ER in a non-monotonic concentration dependent manner. Th e median EEQ of the extracts containing the PFAAs corresponds to the eff ect of 0.5 pg E2 per mL serum. In conclusion, we observed that most of the extracts containing the PFAA mixtures from pregnant women’s serum agonized the ER and enhanced the E2-induced eff ects in non-monotonic concentration-dependent manners.
University of Arkansas for Medical Sciences, USA
Title: Tamoxifen and its derivatives bind to and act at cannabinoid receptors CB1 and CB2 with high affi nity
Time : 10:00-10:20
Anna Radominska Pandya serves as a Professor in the Department of Biochemistry and Molecular Biology at UAMS. She is the Editor in Chief for Drug Metabolism Review. She received her PhD from the Institute of Biochemistry and Biophysics, Polish Academy of Sciences in Warsaw, Poland. She has published 175 papers in various peer-reviewed journals, and has received twelve R01 grants from the NIH and DoD. Her research interests include: The regulation and suppression of human UGTs and their role as anti-proliferative agents in cancer models, the interactions between UGTs and cannabinoid receptors, the delivery of UGT genes and drugs into cancer cells using nanomaterial, and the roles of UGTs in the biotransformation of drugs including resveratrols and drugs of abuse such as marijuana and synthetic cannabinoids.
Tamoxifen (Tam) is classifi ed as a selective estrogen receptor modulator and is chemotherapeutic agent for treatment of estrogen receptor (ER)-positive breast cancer, due to its ability to act as an ER antagonist. We have shown that Tam and its cytochrome P450-generated metabolite 4-hydroxy-Tam (4OH-Tam) also exhibit cytotoxic eff ects in ER-negative breast and pancreatic cancer cells. Th ese observations suggest that Tam and 4OH-Tam can produce cytotoxicity via ER-independent mechanism(s) of action. Cannabinoids compounds have also been shown to exhibit anti-proliferative and apoptotic eff ects in
ER-negative breast cancer cells, and estrogen can regulate expression levels of CBRs. Th is study investigated whether CBRs might serve as novel molecular targets for Tam and 4OH-Tam and we have shown that they bind to CB1 and CB2 with significant affi nity. Furthermore, Tam and 4OH-Tam exhibit inverse activity at CB1 and CB2 in membrane preparations, reducing basal G-protein activity and also act as CB1/CB2R-inverse agonists regulating the downstream intracellular eff ector adenylyl cyclase in intact cells. Th ese results suggest that CBRs are molecular targets for Tam and 4OH-Tam and may contribute
to the ER-independent cytotoxic eff ects reported for these drugs. Th erefore, we hypothesize that the cytotoxicity observed in these cells may be attributed in part to the binding of these drugs to CB1 and/or CB2 causing activation or suppression of downstream genes regulating cell proliferation. If our hypothesis is correct, CBRs could constitute a novel molecular target and structural scaff olds for which eff ective, non-toxic, natural and synthetic cannabinoids might be developed for treatment of various types of cancer.
Banihani S A has completed his PhD from Cleveland Clinic/Cleveland State University, USA in the fi elds Clinical-Bioanalytical Chemistry and Molecular Medicine with full GPA. Currently, he is the Vice Dean of Faculty of Applied Medical Sciences at Jordan University of Science and Technology. He has published more than 25 papers in reputed journals. He has two major research interests: Male Infertility and Clinical Nutrition.
Beta-blockers (or histamine-2 receptor antagonists) are a subtype of acid reducers commonly used to treat the acid-related gastrointestinal diseases (i.e., ulcer, dyspepsia and gastro-esophageal refl ux disease). Even though, these drugs, especially ranitidine and famotidine, are commonly used worldwide, their eff ects on sperm function are still indistinct. This work integratively discusses and summarizes the eff ect of B-blockers on sperm function. The eff ects of nizatidine and ranitidine on sperm function are still controversial. Cimetidine has adverse eff ects on sperm function. In contrast, to date, famotidine does not appear to alter sperm function. Further studies are considered very signifi cant to explain the role of B-blockers on sperm
Inoviem Scientific, France
Title: Individualised clinical toxicology: Physiological intermolecular modulation spectroscopy (PIMS), a technology to forsee drugs efficacy prior to administration
Time : 10:55-11:15
Pierre Eftekhari has completed his PhD from Strasbourg University. He has more than 19 years of experience in Drug Development and is the President of Inoviem
Scientifi c, a company dedicated to cutting edge solutions in drug development. He has published more than 30 papers in reputed journals and filed several patents.
Toxicology today is in need of new insight. In air of individualised medicine the relation between desired clinical eff ect and toxic or side-eff ects should be considered simultaneously. Genomics, proteomics, metabolomics and omics in general, although extremely valuable, are globally lacking clinical relevance. Th erefore, there is a need for new methodologies and new tools enabling us to make a bridge between predicted omics-based drug toxicity and clinics. Here, I shall present Physiological Intermolecular Modulation Spectroscopy (PIMS) a cutting edge techology meant to stratify the patients as responders or non-responders in regard to a pharmacological active agent. PIMS provides individual fi ngerprints based on drug-induced macromolecular modulation directly on human tissue extracts. I will explain the scientifi c background of PIMS and present the results from two diff erent clinical studies (transversal and longitudinal), using peripheral blood mononuclear cells (PBMC) isolated from patients with ulcerative colitis and Crohn’s disease for the prediction of infl iximab effect.
University of Gottingen, Germany
Time : 11:15-11:35
Gratitude is within the ethical tradition of Japanese towards experimental animals, because these provide valuable data for efficacy and safety of pharmaceutical compounds before fi rst administration to humans. Researchers must therefore develop strict responsibilities to use these models under strict adherence to the 3Rs (Refi ne, Reduce, Replace). Models should be relevant; and a relevant model is an animal which expresses receptors/epitopes like humans, reveals comparable pharmacodynamic eff ects and resembles humans in regard to kinetic parameters like metabolism, exposure levels, protein
binding and bioavailability. For primary or secondary pharmacodynamics, next to traditional species, also transgenic or disease models are being used, but for non-clinical safety studies predominantly traditional rodents or non-rodents are tested. The undesirable adverse reactions are oft en so subtle, that for optimal assessments excellent knowledge of specific or spontaneous reactions is needed. Th e model must easily be available, costs acceptable and no paucity of historical data representing an obstacle. There will be a continuation of using rats and mice for toxicity evaluations, but for non-rodents a growing refusal is felt in using dogs and monkeys. Th is paper deals with the utility of the minipigs as an animal model in regulatory toxicology. The advantages or disadvantages will be illustrated. Th e Gottingen minipig is a genetically managed model unlike the dog and monkey toxicology models. Th e basis of the small size of the Gottingen minipig does not involve defective genes. Commercial interests in the pig as an agricultural production species have driven the area of pig genomics. There is no equivalent economic driver for progress in the dog or the non-human primate. The Gottingen minipig
is well positioned for the performance of toxicogenomics studies. Th e close sequence homology between pigs and humans suggest that minipigs could be useful for the testing small molecules but also for biotechnology products.
Th e minipig is the only non-rodent model where transgenic animals can be readily generated, and reproductive technologies are well developed in the pig. Th e biology of the minipig is comparable; practically all study types can be performed in the minipig. For reproductive toxicology studies the minipig off ers numerous advantages although the lack of placental transfer of macromolecules may limit the role of the minipig in reproductive testing of biotechnology products. For safety pharmacology studies the minipig is an advantageous model, particularly as regards the cardiovascular system. Th e immune system of the pig is better characterized than that of the dog and as an omnivore the GI-tract reveals similar characteristics. Overall, the mini-pig should be carefully considered as an alternative to dogs and monkeys; but of course, more comparative data is needed for a rigorous assessment of the usefulness and the predictivity of this species for human drug-induced desired and adverse reactions.
Aarhus University, Denmark
Title: Dioxin-like POPs: Induced aryl hydrocarbon receptor transactivity in the Danish pregnant women
Time : 11:35-11:55
Manhai Long obtained her PhD degree in Medicine in 2007 from Aarhus University and works as Associate Professor of Human Toxicology at the Faculty of Health, Aarhus University, Denmark. She has participated in several international and national projects. She has published more than 30 papers in the international journals and has been serving as peer reviewer of several international journals.
Human exposure to lipophilic persistent organic pollutants (POPs) including polychlorinated dibenzo-p-dioxins/furans (PCDDs/PCDFs), polychlorinated biphenyls (PCBs) and organochlorine pesticide is ubiquitous. Th e individual is
exposed to a complex mixture of POPs being life-long beginning during critical developmental windows. Exposure to POPs elicits a number of species- and tissue-specifi c toxic responses, many of which involve the aryl hydrocarbon receptor (AhR). We aimed to assess the actual level of dioxin-like activity in serum of 806 Danish pregnant women collected during 2011-2013. The bioaccumulated lipophilic serum POPs were extracted by solid phase extraction and clean-up on Supelco multi-layer silica column and fl orisil column. Th e integrated AhR transcriptional activity in the serum fraction was determined using the Hepa 1.12cR mouse hepatoma cell line carrying an AhR-luciferase reporter gene and expressed as pg TCDD equivalent (TEQ) per gram lipid aft er adjusted for the serum lipid. Th e AhR transactivity data was evaluated for possible association to the serum levels of 14 PCB congeners, 10 organochlorine pesticides and/or lifestyle factors. Th e preliminary results showed that 91.3% samples elicited agonistic AhR transactivity. Th e median level of AhR transactivity was 195 pg TEQ/g lipid. Pearson correlation analysis showed a weak positive correlation between dioxin-like activity and PCB 105. No signifi cant correlation between serum dioxin-like activity and pregnant women age, gestational day at blood draw, BMI, smoking status and social economic status were observed.
Ain Shams University, Egypt
Title: Genotypic and phenotypic patterns of antimicrobial susceptibility of Helicobacter pylori strains among Egyptian patients
Time : 11:55-12:15
Marwa Saad Fathi currently works as an Assistant Professor of Medical Microbiology & Immunology at Faculty of Medicine, Ain Shams University. She had her MD degree from Ain Shams University in 2009. She is currently studying a Specialized Diploma in Medical Microbiology at University College, London. She is working as a Director of Medical Mycology Lab at Misr University for Science & Technology (MUST). Her publications exceed 20 papers in important focused journals since 2009.
Backgrounds & Study Aim: Helicobacter pylori is currently recognized as one of the most common chronic bacterial infections worldwide. Eradication of bacteria is eff ective in healing peptic ulcers, preventing ulcer relapses and potentially decreasing the risk of progression to gastric carcinoma. For successful eradication of bacteria, it is imperative that the clinician be aware of
the current antimicrobial susceptibility profi les of isolates within the region. Th erefore, the aim of this study is to compare the phenotypic & genotypic patterns of antibiotics susceptibility to Helicobacter pylori strains among Egyptian patients in order to attain a clinical utility from such patterns.
Patients & Methods: 30 symptomatic cases were enrolled. H. pylori infection was diagnosed by upper endoscopy as well as biopsy was taken. Antimicrobial susceptibility to Helicobacter pylori strains was assessed in all subjects by disc diff usion & e-testing methods. Further molecular characterization for genes encoding antimicrobial resistance of isolated strains was done.
Results: For metronidazole, amoxicillin and ciprofl oxacin, we compared the phenotypic and genotypic patterns of resistance as detected by PCR amplifi cation of the resistance genes. E test results were 100%, 50% & 87.5% for metronidazole, ciprofloxacin & amoxicillin respectively from 16 isolated H. pylori strains.
Conclusion: Improving the knowledge of resistance mechanisms, the elaboration of rational and efficacious associations for the treatment H. pylori infection are of high importance especially in determining the therapeutic outcome. Further progress should ultimately focus on the establishment of a cheap, feasible and reliable laboratory test to predict the outcome of a therapeutic scheme
Institut National de Recherche et de SĂ©curitĂ©, France
Time : 12:15-12:35
Pierre Campo has completed his PhD in Nancy (France) and Postdoctoral studies at Hearing Research laboratory. Currently, he is the Head of Ototoxic and Neurotoxic laboratory at INRS (France). He has published more than 30 publications. He is an Associated Editor at International Journal of Audiology and a Specialist in noise and solvent interactions on hearing. He concieved the EchoScan audio, a new equipment to evaluate the auditory fatigue in factories after a workday.
It is well-known that occupational noise exposure can damage workers’ hearing. It is also well accepted that a combined exposure to noise with cochleotoxic substances such as antibiotics, cisplatin, or chemicals (styrene, toluene and ethylbenzene) can exacerbate the noise eff ects. Although solvent-induced cochlear impairments can be only assessed aft er a long incubation
period, the pharmacological impact of styrene on the central nervous system (CNS) can be rapidly objectifi ed by measuring the threshold of the middle-ear acoustic refl ex (MER) trigger. MER can be precious for preserving the hearing performances of workers. Th e aim of the study was to evaluate the eff ects of a noise (both continuous and impulse) and a low concentration of styrene [300 ppm<(threshold limit value x 10) safety factor] on the peripheral auditory receptor, and on the CNS in rats. The impact of the diff erent conditions on hearing loss was assessed using distortion product oto-acoustic emissions, and
histology studies of cochleae. Although the LEX, 8 h (8-hour time-weighted average exposure) of the impulse noise was lower (80 dB SPL sound pressure level) than that of the continuous noise (85 dB SPL), it appeared more detrimental to the peripheral auditory receptors. If the co-exposure to styrene and continuous noise was less damaging than the exposure to continuous
noise alone, the traumatic eff ects of impulse noise on the organ of corti were enhanced by the co-exposure to styrene. The neuropharmacological eff ects of the solvent explain these surprising results. Actually the CNS eff ects of styrene may account for this apparent paradox. Based on the present results, the temporal structure of the noise should be reintroduced as a key
parameter in hearing conservation regulations.
Beijing Hospital, P.R. China
Liping Yang has completed his PhD from RMIT University in Australia. She is the Vice director of Pharmacy Department, Beijing Hospital, a large-scale general hospitals. He has published more than 90 papers in reputed journals and has been serving as an editorial board member of repute. Since 2003, Dr. Yang has served as an Clinical Pharmacist, Senior Clinical Pharmacist, Associate Professor and Professor for the Pharmacy Department, Beijing Hospital (China). Dr. Yang’s major interests are pharmacogenomics, drug-drug interaction, evidence-based medicine, Chinese Medicine and rational drug use in clinical setting.
What is known and objective: The glycopeptide antibiotics, vancomycin and teicoplanin, are the mainstay of therapy forsevere gram-positive organisms such as methicillin-resistant Staphylococcus aureus. We report a case of Stevens–Johnson syndrome (SJS) induced by sequential therapy with teicoplaninand vancomycin, in a patient with chronic obstructive pulmonary disease (COPD). Case summary: A 74-year-old Han Chinese with 1-year history ofCOPD was admitted for treatment of infective endocarditis. After teicoplanin therapy for 12 days, he developed pruritus andmaculopapular over his trunk and limbs. His rash spread rapidlyto most parts of the body surface area, 7 days after his anti-infection therapy was switched to vancomycin. This was stopped, but he developed SJS when teicoplanin was reintroduced.This patient recovered from his drug eruptions when both teicoplanin and vancomycin were stopped. Pharmacogenetic analyses revealed he was heterozygous with respect to two variants (rs2844682 of MUC21 and rs750332 of BAG6). What is new and conclusion: Cross-reactivity between vancomycinand teicoplanin is rare. SJS attributable to sequential treatment with these two antibiotics has not been reported previously. Care should be taken when prescribing vancomycin in patients with a previous documented skin eruption to teicoplanin, especially in those who carry any susceptibility alleles to SJS/TEN.
INAIL - Department of Occupational and Environmental Medicine, Italy
Pieranna Chiarella has completed her PhD at the department of Histology and Medical Embryology from Sapienza University of Rome and worked as PostDoc at the Univesity of Rome Campus Bio-Medico and as staff member at the European Molecular Biology Laboratory. She is currently researcher at INAIL, Department of Occupational and Environmental Medicine, Epidemiology and Hygiene in Monte Porzio Catone (Rome-Italy).
Several genes encoding for enzymes involved in xenobiotic biotransformation and oxidative stress response have been shown to be polymorphic within the human population. These genes are particularly noteworthy in the occupational toxicology field being useful to identify individuals with higher susceptibility to specific toxic agents, allowing implementation of measures to reduce the exposure risk. Although blood is the first choice biological matrix to extract genomic DNA, there are some critical issues hindering its use in occupational studies: 1) blood draw invasiveness 2) difficulty to achieve consent from donors 3) need of the authorized Medical Doctor for blood sample collection 4) safety issues during blood processing 4) time-lenght of cell isolation procedure. Since urine is the routine sample used in the biomonitoring of occupational exposure to chemical agents, the use of the same sample would be advantageous for both purposes, i.e. biomarker determination and genotyping. Here we report an effective method to isolate genomic DNA from urinary sediment. The protocol is not based on use of commercial kit, is cheap, rapid and allows to determine the genotypic status of individuals using PCR and RFLP analysis. Before applying this procedure to exposed workers, we tested the efficacy on urine harvested from our Institute donors and carried out genotyping analysis of GSTT1 and GSTM1 enzymes. In conclusion, at least for the genotyping analysis, urine sediment represents a suitable alternative to whole blood. This approach might be extended further to identify specific gene and protein biomarkers of dose/exposure in the occupational and environmental setting.
JSS University, India
Dr.J R Kumar, Ph.D in Biochemistry from University of Mysore in 2008 (India) subsequently joined as postdoctoral fellow in Medical college of Wisconsin, Milwaukee and UND school of medicine, Grandfork, USA, Area of research includes snake venom pharmacology and remyelinating strategy in multiple sclerosis. Reviewer and editorial member for more than 10 reputed international journals. Young Scientist award from the Department of Science and Technology, government of India.
The snake venom is an mixture of toxic and nontoxic proteins. The toxic proteins include enzymatic and non enzymatic proteins. The main toxic enzymes are PLA2s and PLA2 containing toxic complexes, hemorrhagic proteases and hyluradinases. Among all enzymes PLA2s are well studied in terms of structure, function and mechanism of pharmacological action. Most of toxic PLA2s are basic in nature and target specific. The well studied viper snake venom PLA2s from Indian origin are VRV-PL-V, VRV-PL-VIII, VRV-PL-VI, VRV-PL-VII and VRV-PL-IX. The VRV-PL-V and VRV-PL-VIII are major toxic PLA2 from southern region Russell’s viper venom. VRV-PL-V is neurotoxic and VRV-PL-VIII induces hemorrhage in the lungs. VRV-PL-VI from northern region India causes hemorrhage in pituitary gland and peritoneal cavity. VRV-PL-VII and VRV-PL-IX are neurotoxic PLA2 from western region India. VRV-PL-IX is presynaptic, VRV-PL-VII and VRV-PL-V (Southern, India) are exhibited pre and post synaptic neurotoxicity in cultured hippocampal neurons through NMDA and nonNMDA receptors. This is the first report from the snake venom PLA2s having both pre and post synaptic neurotoxicity. Furthermore A novel heterotrimeric toxic complex “Reprotoxin” isolated from western region Russell’s viper venom, having PLA2, protease and trypsin inhibitor peptide. It is target specific to Male and female reproductive system affecting Leydig and sertoli cells in males and Follicles in females.
Dicle University Faculty of Veterinary Medicinei Diyarbakir, Turkey
I have completed my PhD at the age of 31 years from Yuzuncu Yil University School of Veterinary Medicine. I have published more than 4 papers in reputed journals.
The aim of this study was to investigate the antioxidant properties and protective role of honey, considered a part of traditional medicine, against carcinogen chemical aflatoxin (AF) exposure in rats, which were evaluated by histopathological changes in liver and kidney, measuring level of serum marker enzymes, antioxidant defense systems, and lipid peroxidation content in liver, erythrocyte, brain, kidney, heart and lungs. For this purpose, a total of eighteen healthy Sprague-Dawley rats were randomly allocated into three experimental groups: A (Control), B (AF-treated) and C (AF+honey-treated). While rats in group A were fed with a diet without AF, B, and C groups received 25 µg of AF/rat/day, where C group additionally received 1 mL/kg of honey by gavage for 90 days. At the end of the 90-day experimental period, we found that the honey supplementation decreased the lipid peroxidation and the levels of enzyme associated with liver damage, increased enzymatic and non-enzymatic antioxidants in the AF+honey-treated rats. Hepatoprotective and nephroprotective effects of honey is further substantiated by showing almost normal histological architecture in AF+honey-treated group, compared to degenerative changes in the liver and kidney of AF-treated rats. Additionally, honey supplementation ameliorated antioxidant defens systems and lipid peroxidation content in other tissues of AF+honey-treated rats. In conclusion, the present study indicates that honey has a hepatoprotective and nephroprotective effect in rats with experimental aflatoxicosis due to its antioxidant activity.
Introduction and Objective: In the modern world, soy bean as a valuable meal is used for providing protein and some essential body needs. The stomach also as a part of alimentary canal has its own role in mechanical and chemical digestion. The objective of this study is evaluation of the effects of soy meal on histological and histometrical features of the stomach and also on biochemical factors of blood serum.
Methods: In this experimental study, a total of thirty immature female mice of Balb/C were selected at three weeks of age. Then on the basis of three types of diets, the mice were divided into three groups as; the group A or control, fed on a diet of full protein, the group B fed on a diet of 40% soy meal and the group C fed on a diet of 20% soy meal. After three months, the mice were anesthetized, and blood samples were obtained from the heart for determining the serum level of some hematologic factors such as urea, total protein, cholesterol and LDL. Then the stomach was removed, some tissue sections were prepared and stained with H&E. After histological study, the stomach was subjected to histometric evaluation. The histometric data were surveyed by a light microscope equipped with Axiovision software and the thickness of mucosa, submucosa, musculature, also depth of the pits and the number of parietal cells were measured. For data analysis, One Way ANOVA was used to compare the control group with experimental groups and Tukey test was used to compare the groups with each other. The significance level was considered as P<0.05.
Findings: Soya bean consumption didn,t cause histological changes.In the more precisely histometric study, the results showed that in non glandular portion of the stomach, between the control group compared with the groups fed on soy meal, there was significant increase in thickness of mucosa in the experimental group B (676±99.45) compared with the control group (427±77.53) and also in thickness the muscular layers in the experimental group C )233.30±84.69) compared with the control group (104.05±11.71), (P<0.05). In the glandular portion also significant increase in thickness of mucosa in the experimental group B (1041.36±167.02) and C (1331.73±143.32) compared with the control group (615.29±83.14), in depth of pits in the experimental group B 134.53±14.60) and C (154.29±20.25) compared with the control group (94.79±12.93), in parietal cells in the experimental group B (15.66±4.45) and C (30.50±13.61) compared with the control group (9.83±1.47) and muscular layers in the experimental group B 211.59±53.68) and C (195.72±67.89) compared with the control group (155.73±28.23) was observed (P<0.05). However a significant decrease in serum level of cholesterol in the experimental group B (87.76±20.53) and C (83.03±16.06) compared with the control (117.70±20.71), serum level of urea in the experimental group B (36.23±10.43) and C (40.58±12.14) compared with the control group (65.33±7.28) and also LDL value in the experimental group B (5581.6±470.90) and C (5689.1±479.79) compared with the control group (6907.1±37.64) was observed (P<0.05).
Conclusion: It seems that long term consumption of soy bean could affect on the stomach mucosa and proliferation of parietal cells and also could decrease the serum levels of cholesterol, urea and LDL.
- Young Researchers Forum
Location: Rome, Italy
BioMath GmbH, Germany
Title: Proposed criteria for the evaluation of the scientific quality of mandatory rat and mouse feeding trials with whole food/feed derived from genetically modified plants
Time : 13:20-13:40
Kerstin Schmidt completed her University degree in Mathematics with specialisation in Statistics and Probability Theory. In 1990, she established her own company BioMath, an internationally operating consultancy for research institutions and industrial partners in statistics and informatics, especially in the life sciences. She has been accompanying more than 100 projects in Toxicology, several of them joint/ international projects. She works as a Lecturer for Statistics and Experimental Design at the University of Rostock. In April 2016, she submitted her Doctoral thesis entitled “Statistical aspects and methods of the risk assessment and postmarket environmental monitoring of genetically modified plants”.
In recent years, animal feeding trials conducted with whole food/feed have been a focal issue in the controversy on the safety assessment of genetically modifi ed (GM) plants and derived food/feed. Within the scientifi c community and among stakeholders, quite diff erent views have been expressed on how these studies should be conducted, analysed and interpreted, what they might add in terms of information relevant to safety and whether 90-day rodent feeding trials should be mandatory. In the context of the ongoing debate on GMO risk assessment in Europe, it is crucial to investigate: Criteria for evaluating the scientific quality of subchronic, chronic toxicity and carcinogenicity studies with whole food/feed in rats and mice. This will
help risk assessors in evaluating this type of studies when provided in the course of a pre-market risk assessment and will create a basis for further general debate. Th is talk specifi cally addresses the question on how to evaluate whole GM food/feed feeding trials. It does so by proposing a list of key quality criteria for the evaluation of 90-day and extended feeding trials with whole food/feed derived from GM plants. Th e proposed quality criteria should be taken into account when evaluating a feeding trial in the frame of an application to regulatory bodies and are not intended to be applied in other cases in which a feeding trial is performed to answer a specifi c open question in basic research.
University of Birmingham, UK
Title: The effect of the first- and second-generation of antipsychotic drugs on SH-SY5Y brain cells and their toxicity
Time : 13:40-14:00
Israa J Hakeem is a PhD student at the University of Birmingham. She has completed her Master’s degree in Forensic Science from Anglia Ruskin University and received Bachelor’s degree in Biochemistry.
Antipsychotic drugs are primarily used to manage several psychiatric disorders, including schizophrenia, bipolar mania and related mental illnesses. Th e present study examined the eff ect of the fi rst and second generation of antipsychotic drugs on neuronal and non-neuronal cells. Th e toxicity of both-generation of antipsychotics was tested in both the SH-SY5Y brain cell line and the COS7 kidney cell line. According to the LC50 values for chlorpromazine (1st generation), Trifl uoperazine (1st generation) and Olanzapine (2nd generation), the neurotoxicity of the two classes in SH-SY5Y exceeded their common cytotoxicity in COS7 cells, indicating that neuronal cells are at greater risk of cell death with low concentrations of antipsychotics
at micro-molar comparing to non-neuronal cells. Detailed studies looking at the mechanisms of cell death induced by these antipsychotic drugs indicate that both apoptosis and necrosis play a role, while autophagy does not.
Banasthali University, India
Time : 14:00-14:20
Sonam Agarwal is pursuing PhD in Biotechnology from Department of Bioscience and Biotechnology, Banasthali University, Rajasthan, India. She has completed her MSc in Biotechnology from MITS University, Rajasthan, India. Her research work highlights “The role of bacoside A and bromelain against dichlorvos incited toxicity in serum and kidneys of mice”.
Current study emphasizes the toxic eff ects of dichlorvos on serum in terms of oxidative stress. Meanwhile, a protective action of bacoside A and bromelain was investigated against the biochemical alterations in serum. Experimental design included six groups of mice: Saline was given as a vehicle to the control mice (group I). Mice belonging to groups II, III and IV were administered with dichlorvos (40 mg/kg b.w.), bromelain and bacoside A, respectively. Fift h group received a combination of bromelain and bacoside A. In group VI, bacoside A and bromelain were administered 20 minutes prior to exposure of dichlorvos. Th iobarbituric acid reactive substances (TBARS), protein carbonyl content (PCC), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) level were used as biochemical test of toxic action for dichlorvos intoxication. Signifi cantly increased TBARS and PCC level in second group suggests that dichlorvos enhances the production of free radicals in serum of mice (p<0.05). Antioxidants treatment signifi cantly decreased the levels of TBARS and PCC (p<0.05). Dichlorvos administration causes a signifi cant reduction in the level of CAT, SOD, GPx and GSH (p<0.05) which was restored signifi cantly by co-administration of bromelain and bacoside A in dichlorvos exposed mice (p<0.05). Treatment of bromelain and bacoside A in combination served as better scavengers of toxicity induced by dichlorvos.
CHU Bab el Oued, AlgĂ©ria
Title: Determination of chromium by ETAAS in hair and urine of tannery workers: The interest of alternative biological matrices
Time : 14:20-14:40
Mohammed Riffi has completed his PhD from Tlemcen University and Post-doctoral studies from Algiers University School of Medicine. He has a membership of SoHT. He has published one paper in Toxicologie analytique et Clinique.
Introduction: Th e human hair occupies a prominent place as markers of exposure to xenobiotics in the domain of forensic toxicology. However, interest of using this matrix is a considerable improvement for the assay of metals; also, chromium has attracted the attention of toxicologists because the accidents observed in industrial settings using this metallic element (cement
industry, paint, leather, automotive, etc.).
Objective: Th is study is related to the assessment of worker exposure in a tannery, located in Algiers, more accurately on the Rouiba-Reghaia industrial estate, by measuring the capillary and urinary chromium of the population groups investigated, and then study the correlation between total content of chromium in hair and urine.
Patients & Methods: Th e study was carried out in September 2012, and focused on 50 subjects exposed (49 men and 1 woman) and 16 controls. It was preceded by the establishment analytical development and validation of an analytical method for the determination of chromium in hair and urine by graphite furnace atomic absorption spectrometry (GFAAS). Statistical
calculations were performed using the soft ware LXSTAT MS Excel 2012.
Results & Discussion: Th e hair chromium average of the tanning workers were signifi cantly higher (urinary Cr=2.48 μg/L, Cr capillary=4.93 ng/mg) than other groups. Washing the hair appears to be eff ective for decontaminating the exogenous chromium, this latter may refl ect a recent exposure to chromium.
Conclusion: Human hair may off er the advantage for biological monitoring, fi rst, to get information on the use of means of
protection, and give some idea about the route of exposure (inhalation or ingestion).
Lena Schober completed her Engineering degree in Biotechnology from University of Applied Sciences Esslingen in 2009. Since 2009, she is working as a Research Fellow for the Fraunhofer Society. She owns particular knowledge and practical experience in Cell and Tissue Engineering and focuses on the “Development of automated cell application systems for research and clinical use”. She thereby contributed to the Fraunhofer Project “Mass customized organ replicates-tissue engineering on demand” and advanced the transfer of the biological process to the automated system, also referred to as “tissue factory”, and performed the validation of produced in vitro systems.
The use of automated systems for biological tasks is in great demand. Nevertheless, there are a lot of barriers at the moment limiting the successful application of automated systems. By the lack of fl exibility and the demand for skilled computer scientists & engineers just the two main aspects stated by experts shall be mentioned. Th e Fraunhofer IPA has a strong background on automated cell culture technologies. Th e expertise, gained in the successful “tissue-factory” light-house project, let us rethink the overall process chain and overcome established principles. A concept that has a strong link to current industry 4.0 concepts and applies a seamless integration throughout the value-added chain will be presented. The main idea is to provide maximal transparency through digitalization and the design of smart automates. Th e interface to the customer-pharmaceutical company, regulatory board or consumer is the disruptive change with regard to state of the art attempts. Starting with the vision of full transparency down to the bench, the overall infrastructure and test processes need to be reconsidered.
Gorgan University of Agricultural Sciences and Natural Resource, Iran
Time : 15:00-15:20
The main aim of the present study was to examine the sensibility of marine fi sh Mugil cephalus to dichromate potassium (K2Cr2O7) in the toxicity test programs. All fi shes were exposed to (K2Cr2O7) at various chosen concentrations 0, 5, 10, 20, 30, 40, 50, 55 ppm (range fi nding test). Th en, fi sh were exposed to 6 concentrations of (K2Cr2O7) (control, 60, 70, 80, 90, 100 ppm). Number of mortality was registered aft er 24, 48, 72 and 96 h. LC50 values were determined with probite analysis. The 96 hour LC50 value of (K2Cr2O7) to the fi sh was found to be 83.07 ppm. By comparing the sensitivity of this metal to common reference toxicants, we conclude that grey mullet can be used as a suitable model for toxicity determinations in ecotoxicological studies. Further studies should examine other contaminants of this species to assess their suitability for detecting toxicity, as well as complex mixtures of pollutants, in order to develop aquatic ecosystem monitoring programs.
Zeinab H Arabeyyat has completed her MSc in Biotechnology from Jordan and is currently a fi nal year PhD student in Aquatic Toxicology Research Group at the University of Hull in UK. She has attended many international conferences in Norway and local conferences in Jordan and UK. She is a Lecturer in the Faculty of Marine Sciences at the University of Jordan.
It is important to develop early warning tools of nanoparticle-induced biological eff ects for aquatic species to be able to monitor any possible impacts. In this study, early zebrafi sh (Danio rerio) embryos have been experimentally exposed in vitro to 1.925 mg/L of 4 nm, 10 nm AgNPs and to silver ions (0.018 mg/L) alone, up to 96 hpf. Five targeted genes have been
employed for analysis: Peroxisomal membrane protein 2 (Pxmp 2), hypoxia inducible factor (HIF), superoxide dismutase (SOD), mucosal secretion protein (Muc) and catalase (CAT) genes. A global approach employing suppression subtractive hybridization (SSH) has also been used in parallel to identify novel genes that may be involved in the fi sh embryo response as a result of exposure to nanoparticles. Th e results show that 4 nm AgNPs are taken up by zebrafi sh embryos at a concentration of 1.925 mg/L. AgNP uptake resulted in signifi cantly up-regulated Pxmp 2 and HIF mRNA transcript levels in exposed embryos. An increased trend in up-regulation of SOD was also observed, while, Muc and CAT remained unchanged. No corresponding significant differences were observed in any of the transcript levels analyzed following exposure to larger sized 10 nm AgNPs or silver ion exposure alone. An up-regulation of solute carrier family 25, membrane 5 and Cytochrome c oxidase subunit I; and down-regulation of spermatogenesis associated protein 2 and Actin alpha, cardiac muscle lb mRNA expressions identified by SSH approach have also been observed. Th ese results suggest that 4 nm AgNPs are available for uptake and, as a result cause changes in mRNA expression in developing embryos.
Universidade Federal do Rio Grande, Brazil
Title: Anti-inflammatory effect and toxicology analysis of oral delivery quercetin nanosized emulsion in rats
Time : 15:40-16:00
Gabriela Hädrich is pursuing her PhD from Federal University of Rio Grande, Brazil. She is a visiting PhD student at Martin-Luther Universität Halle Wittenberg, Germany. She has completed her Master degree in Health Sciences in 2014 and has expertise in Nanotechnology applied to health and microbiology.
This study evaluates the advantage of the quercetin encapsulation in nanosized emulsion (QU-NE) administered orally in rats in order to demonstrate its anti-oedematous and antioxidant eff ects as well as its toxicity. The nanocarriers were prepared using the hot solvent diffusion with the phase inversion temperature methods. The nanocarriers physicochemical properties were then investigated. The anti-edematous activity was tested using paw edema in rats. In addition, NF-kB expression in subcutaneous tissue of the paws was accessed by immunohistochemistry while the lipid peroxidation was
analyzed in the liver by malondialdehyde reaction with thiobarbituric acid. Hematological, renal and hepatic toxicity as well as the genetic damage were also evaluated. Th e results demonstrated that QU-NE exhibited pronounced anti-oedematous property comparable to drug diclofenac. This eff ect was associated with NF-κB pathway inhibition. Th e lipid peroxidation was also only reduced in rats treated with QU-NE. Besides this, no genetic damage, hematological, renal or hepatic toxicities were observed aft er administration of QU-NE. These results suggest that quercetin nanosized emulsion exhibits anti-oedematous and antioxidant properties and does not demonstrate toxic effects. Th is indicates that it has a potential application in the treatment of inflammatory diseases.