Effects of pesticide on the thyroid hormones of pesticide sprayers living in Mazandaran

Abstracts Oral – [A-10-476-1] Role of glutathione system in biochemical and metabolic functions of hepatocytes-like cells differentiated from human bone marrow mesenchymal stem cells Abdolamir Allameha, Ahmadi-Ashtiani Hamidrezaa, Kazemnejad Somaieha, Soleimani Masouda, Gharebaghian Ahmadb, Rastegar Hosseinc, Ismaeili Shahnazb, Hedayati Mehdib a POB 14115–111, Tehran, Iran b Tehran, Iran c Ministry of Health, Tehran E-mail address: allameha@modares.ac.ir (A. Allameh) Introduction: Glutathione plays a major role in various cellular functions. However our information about glutathione system involvement in biochemical and metabolic functions of hepatocyte-like cells during differentiation from mesenchymal stem cells is scanty. This study is focused on the role of glutathione on biological activity of hepatocytes during differentiation. Methods: Differentiation of MSCs into hepatocytes was induced using standard protocol in a specific culture media. The non-differentiated and differentiated cells were collected at different time intervals (0, 2, 7 and 14 days) and the rate of cell viability (Trypan blue test) and proliferation (Brdu assay) were monitored. After characterization of hepatocytes and their progenitor MSCs, the levels of cellular albumin, alpha-fetoprotein (AFP), cytochrome P450 (CYP), reduced glutathione (GSH), oxidized glutathione (GSSG), total glutathione, GSH/GSSG ratio, GSH peroxidase (GPX) and GSH reductase (GR) were measured and compared to respective controls. Albumin and urea and secretion into the culture media were monitored as the biosynthesis capacity of the hepatocytes. Results: The growth curve of the differentiated hepatocytes revealed that the MSCs were successfully differentiated and the proliferation rate was normal during three-week culture. Expression of liver function factors such as; albumin, AFP and CYP were increasing following hepatogenic differentiation of the MSCs. It was also found that glutathione modifiers can influence cell function and metabolism. Excretion of urea and albumin into the culture media as index of metabolic activity was evident on day 7 of differentiation. Treatment of the cells with glutathione modifiers showed that the cellular GSH play a major role in cell survival and biological activity. Conclusions: Cellular glutathione as a major antioxidant factor could play a major role in viability, longevity and metabolic function of hepatocytes differentiation from adult stem cells. Keywords: Biochemical, Metabolic, Hepatocytes, Stem cell, Differentiation doi:10.1016/j.clinbiochem.2011.08.1061 Oral – [A-10-1458-2] Promoting chemical safety through toxicology education in the classroom Rahmat Awanga, Nida Besbellib, Agneta Sunden Bylehnc, Birger Heinzowd, Lillian Corrae a National Poison Centre, Universiti Sains Malaysia, Malaysia b United Nations Environment Programme, Geneva c LGA-SH, State Agency for Health of Land Schleswig-Holstei, Germany d New Zealand Poison Centre, New Zealand e International Society of Doctors for the Environment, Argentina E-mail address: rahmatawang@yahoo.com (R. Awang) In many developing countries, infants and children are particularly vulnerable to pesticides and other toxic chemicals because their bodies are smaller and still developing. Children also face greater S43 exposures than adults due to their hand-to-mouth behaviors. Children living in farming areas or whose parents work in agriculture suffer greater pesticides exposure than other children. Non-chemical methods exist to prevent or manage pests. Integrated Pest Management (IPM) and Integrated Vector Management (IVM) apply different methods, trying to reduce the use of chemicals to a minimum. Despite this, pesticides are much used in farming economies today, and for managing vectors of disease, especially in developing countries and countries with economies in transition. Pesticides are applied to kill pests, but they can also adversely affect non-target organisms, including humans. In the developing world most people live in rural areas and children's exposure to pesticides is unavoidable since farms are homes as well as workplaces. Educational materials that is aimed specifically at creating awareness among young students about potential side effects of chemicals and help reduce careless use and handling of pesticides and other chemicals are not available. This has brought the attention of the United Nations Environmental Programme (UNEP) and the World Health Organisation Regional Office for Europe (WHO Euro) and the Malaysian National Poison Centre of Universiti Sains Malaysia (MNPC) with initial support from the International Union of Pure and Applied Chemistry (IUPAC) to develop an innovative teaching courseware utilizing multimedia that will help provide basic understanding of toxicology and awareness of the need for protective and precautionary measures to minimise adverse effects on human health and the environment. This paper highlights how young children learn toxicology and interact with the courseware. The experience gained through its testing in Malaysia and Argentina will also be described. doi:10.1016/j.clinbiochem.2011.08.1062 Oral – [A-10-818-1] Effects of pesticide on the thyroid hormones of pesticide sprayers living in Mazandaran Fatemeh Tohidia, Farzaneh Farrokhib, Ali Taravatic a Department of Modern Sciences and Technologies Mashhad University of Medical Science Mashhad, Iran b Islamic Azad University, Sari Branch, Sari, Iran c Islamic Azad University, Jouybar Branch, Jouybar, Iran E-mail addresses: tohidiF891@mums.ac.ir (F. Tohidi), ali_taravatie@yahoo.com (A. Taravati) Introduction: Pesticides are occasionally used indiscriminately in large amounts causing environmental pollution. Pesticides cause adverse effects on different body systems, including hematological and endocrine systems. The ability of organophosphate pesticides to disturb thyroid gland function has been demonstrated by experimental studies on animal, but evidence of such effects on human remains scarce. The purpose of this study was to investigate the association between exposure to pesticide and serum levels of thyroid hormones in pesticide sprayers. Methods: Blood samples were obtained from 40 cases and 20 controls after overnight fasting. Glucose, urea, acid uric and lipid profile were measured by enzymatic methods. TSH, T3 and T4 levels were determined by immunoassay kits. Results: The TSH level in sprayers (2.64 ± 1.50 μM, P = 0.007) was significantly increased compared with the control subjects (1.33 ± 0.83). T3 and T4 levels of sprayers were lower than controls. Conclusion: These results suggest that exposure to organophosphate pesticides may be responsible for increasing TSH level and decreasing T3 and T4 serum hormone levels, therefore supporting the hypothesis that organophosphate pesticides cause hypothyroidism in S44 Abstracts pesticide sprayers. So administration of supplementary nutrients or improving defense system in these subjects is advised. Keywords: Hypothyroidism, Pesticide, TSH, Immunoassay doi:10.1016/j.clinbiochem.2011.08.1063 Oral – [A-10-831-1] Protective role of N-acetyl-cysteine on diazinon induced oxidative stress in rat kidney Farideh Izadia, Mahvash Jafarib, Alireza Asgarib, Maryam Salehib a Department of Biology, Islamic Azad University, Tehran, Iran b Baqiatall University, Iran E-mail addresses: Izadi_FArideh@yahoo.com (F. Izadi), jafari@bmsu.ac.ir (M. Jafari) Introduction: Organophosphate pesticides (OP) may cause generation of reactive oxygen species (ROS), which may lead to oxidative stress. Diazinon (DZN) is an organophosphorus insecticide that has been widely used throughout the world with applications in agriculture and horticulture for controlling insects. N-acetyl-l-cysteine (NAC), precursor of reduced glutathione (GSH), is an antioxidant and free-radical scavenger. In this study, the possible protective role of NAC in DZNinduced toxicity was evaluated in male Wistar rats. Materials and methods: In vivo experimental groups were: control group (corn oil as diazinon solvent), DZN group (100 mg/kg, i.p.), NAC (160 mg/kg, i.p.) group and NAC–DZN group. 24 hours after injection, animal to ether anesthesia and kidney removed and superoxide dismutase (SOD) and catalase (CAT) activities, as well as GSH and malondialdehyde (MDA) levels were determined by biochemical methods. Results: The result showed that SOD and CAT activities and MDA level were increased, while GSH level was decreased in DZN treated rats as compared to control. NAC–DZN groups were found to improve these disorders. Conclusion: The results suggest that NAC provides protection against DZN-induced oxidative stress and prevents lipid peroxidation in kidney. Keywords: Diazinon, N-acetyl-cysteine, Antioxidant system, Rat, Kidney doi:10.1016/j.clinbiochem.2011.08.1064 Oral – [A-10-1001-1] Activation of Nuclear Factor-kappaB by doxorubicin leads to inhibition of extrinsic pathway of apoptosis in H9c2 cells Leila Hosseinzadeh, Gholamreza Karimi, Fatemeh Mosaffa, Mitra Rezaee Asgharian Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran E-mail addresses: L_hoseinzadeh@yahoo.com (L. Hosseinzadeh), gkarimi@mums.ac.ir (G. Karimi), ft_mosaffa@yahoo.com (F. Mosaffa) Introduction: Doxorubicin (DOX), a chemotherapeutic agent, can give rise to serve cardiotoxicity by inducing apoptosis. The exact mechanism of DOX-induced apoptosis has not been fully understood yet. Here, we evaluated the effect of DOX on some important genes involved in apoptosis and also Nuclear Factor-kappaB (NF-κB) activity in H9c2 cardiomyoblast cells. Materials and methods: Cell viability was determined by MTT. Apoptosis was assessed by annexine V/PI double staining. Real time RT- PCR was used to evaluate the expression of genes. The DNA-binding capacity of NF-κB was examined by ELISA. Results: DOX-mediated cytotoxiocity is executed by inducing apoptotic cell death. DOX induced apoptosis by down regulating antiapoptotic Bcl-2 and up-regulating pro-apoptotic Bax. Consequently, the ratio of Bax to Bcl-2 is significantly increased upon treatment with DOX leading to increase caspse-9, the initiator caspase of mitochondrial pathway for apoptosis, expression. However, it had no effect on the expression of caspase-8 the mediator of extrinsic pathway. DOX wasn't able to inhibit expression of anti-apoptotic gen cIAP1. Also, we observed that NF-κB activity was sharply increased by incubation in the presence of DOX. Previous studies have shown that depending on the cell models, apoptosis from DOX can follow different pathways. Our findings demonstrated that apoptosis induced by doxorubicin occurred through intrinsic pathway rather than by extrinsic pathway. It was indicated that NF-κB induces multiple factors to regulate apoptosis including cIAP1. Conclusion: Based on these results, it is possible that NF-κB activation suppresses extrinsic pathway of apoptosis through blocking the cIAP1 down-regulation and consequently caspase-8 activation by DOX in H9c2 cells. Keywords: Doxorubicin, NF-kappaB, Apoptosis, H9c2 cells doi:10.1016/j.clinbiochem.2011.08.1065 Oral – [A-10-1026-2] Apoptosis induction of methysulfonylmethane in human gastrointestinal cancer cell lines Naser Jafari, Shahab Bohlooli Ardabil University of Medical Sciences, Ardabil, Iran E-mail address: n.jafari@arums.ac.ir (N. Jafari) Introduction: Methylsulfonylmethane (MSM) is an organic sulfur containing compound that occurs naturally in a variety of fruits, vegetables and unpasteurized milk. It has been used as a supplement in the last two decades. Recently it has been suggested that MSM has clinical potential as a non-toxic agent effective against metastatic melanoma. Considering the preventive effects of this substance on tumor onset and nontoxic to healthy body we have investigated in vitro effects of methylsulfonylmethane on apoptosis induction in human gastrointestinal cancer cell lines. Methods: The human cancer cell lines including, AGS, HepG2 and KYSE30 were cultured and incubated until confluence. The cells were removed and seeded in 96-well plates at a density of 1 × 103 cells/well and incubated overnight, and then treated with (21–30) mg/ml MSM. To stain apoptotic cells, the plates were centrifuged. The EB/AO dye mix was prepared in PBS and 10 μl was added to each well and cells were viewed and counted under an inverted fluorescence microscope. Results: The results showed that after incubation at (21–29) mg/ml of MSM for 24 h the cells displayed a series of morphological changes including condensation and fragmentation of chromatin and nucleus, and formation of apoptotic bodies which were designated as typical evidence of apoptotic bodies. In contrast control cells exhibited a normal appearance. There was a significant increase of apoptotic cells in cancer cells after treating with MSM. Conclusion: The results demonstrated that MSM exerts its anticancer and cytotoxic effect by inducing apoptotic cell death. Keywords: Apoptosis, Methylsulfonylmethane, Cancer doi:10.1016/j.clinbiochem.2011.08.1066