Elif Karacaoğlu

Toxicity of food contaminant furan on liver and kidney of growing male rats

Furan is a chemical used in some industrial products and occurs naturally in heat‐treated foods. We aimed to investigate the effects of orally administered furan on liver and kidney in growing Wistar male rats for 90 days. In this respect, biochemical, morphological, histopathological, and histomorphometrical examinations were performed. Three‐ to 4‐week aged rats were divided into five groups of eight animals each; control, oil control; 2, 4, 8 mg/kg/day furan treatment groups. At the end of the experiment, antioxidant enzyme activities and serum AST, ALT, HDL, Urea, etc. levels were analyzed. Malondialdehyde (MDA) levels, superoxide dismutase (SOD), catalase (CAT), tumor necrosis factor‐α (TNF‐α), and interleukin‐6 (IL‐6) were also measured in liver homogenates. Also, liver and kidney were examined morphologically and histopathologically under light microscopy. According to the results of biochemical analysis, ALT, ALP, and LDL levels in treatment groups were significantly different compared with control groups. While LDL levels in treatment groups increased significantly, ALT and ALP levels decreased significantly. No significant changes were observed in liver MDA levels, superoxide dismutase and catalase activities in treatment groups. While IL‐6 levels did not change in treatment groups, furan caused dose‐dependent increases in liver TNF‐α level of rats. In treatment groups, absolute and relative liver weights changed significantly, however, no significant changes were observed in kidney and relative kidney weights. Hyperemic blood vessels in the liver and congestion, edema, fibrosis, and tubular damage in the kidney of rats treated with furan were observed histopathologically. According to histomorphometric examinations, glomeruli diameters and glomerular volume decreased in the kidneys of rats in treatment groups.

Effects of heat-induced food contaminant furan on reproductive system of male rats from weaning through postpuberty.

Furan (C(4)H(4)O) is a volatile, colorless liquid and is used in some segments of the chemical manufacturing industry. It is found in variety of foods such as coffee, jarred and canned foods that undergo heat treatment. This study was designed to investigate the effect of furan exposure on reproductive system of male rats. Three to four weeks old rats were exposed to furan at 2, 4 and 8 mg/kg/day doses by orally for 90 days. Hematology, weights, histology and morphometry of reproductive organs, serum LH and testosterone levels, sperm count and morphology and apoptosis in testis were evaluated. Slight changes were observed in hematological parameters of furan-treated rats. The weights of seminal vesicle reduced significantly whereas the weights of prostate increased significantly in the highest furan dose group. LH and testosterone levels decreased in furan-treated rats. Histological examinations have revealed that furan caused impairments in testis, epididymis and prostate gland. Furan showed no effects on sperm counts and morphology. On the other hand apoptotic cells in testis increased significantly. According to morphometrical examination, the epithelial heights and lumen diameters of the reproductive organs have changed in treatment groups. These results indicate that subchronic furan treatment induces toxicity of the male reproductive system.

Does furan affect the thymus in growing male rats

Furan has been identified in foods such as heat-treated foods, including coffee, canned meat, hazelnuts, and infant foods and formulas. Children may be exposed to furan via either consumption of these foods or their derivatives. We evaluated the effects of furan on the thymus of weaning male rats in the present study. Five separate groups containing male rats were used: control, oil control, and three furan-treated groups. Furan was given orally to rats in the treatment groups at doses of 2, 4, and 8 mg/kg/day for 90 days. At the end of the experiment, thymus of the rats were examined morphologically, histopathologically, and immunohistochemically. We observed that absolute and relative weights of thymus were decreased significantly in rats treated with 4- and 8-mg/kg/day doses of furan. In histopathological examination, enlargement of interstitial connective tissue between the thymic lobules, lymphocyte depletion, and hemorrhage were observed. We detected an increase in apoptotic cell counts in thymus of the treatment groups. In addition, we found significant differences in the distribution of fibronectin and transforming growth factor-beta in the thymus of the treatment groups. In conclusion, we suggest that furan has affected the thymus in growing male rats.

In vivo toxicity of a new antifungal agent 2,4-dithiophenoxy-1-iodo-4-bromo benzene: a follow up on our in vitro study

Abstract Triazole fungicide fluconazole has become the most widely used antifungal agent in the world, mainly because of its ability to penetrate well into body fluids and tissues. However, it has been reported to interact with many drugs and because of its common use, the risk of resistance to fluconazole increases. This calls for new anti-fungal drugs that would be able to replace it. In 2006, a new thialo benzene derivative - 2,4-dithiophenoxy-1-iodo-4-bromo benzene (C18H12S2IBr) - was synthesised with a carbon backbone similar to fluconazole, and, according to the early in vitro tests, much greater efficiency. Followed an in vitro test of its cytotoxicity, in which the new drug showed promising results as an alternative to fluconazole. The aim of this study was take the next step and test C18H12S2IBr toxicity in vivo. We opted for a four-week test on Wistar rats, in which the new antifungal agent was orally applied at doses two and a half and five times lower than those of fluconazole. There were no changes in daily food and water consumption, but weight gain in female rats and relative organ weights changed in the treated groups, pointing to sex-related differences in drug metabolism and effects. Fluconazole significantly increased leukocytes and lowered neutrophils whereas C18H12S2IBr did not, while other haematological changes in respect to the vehicle control were similar between the treated groups. Differences in cytochrome c in the liver and kidney suggested greater apoptotic effect of the new drug, but interpretation remains inconclusive, considering that other key indicators (biochemistry and histopathology) do not support greater toxicity. Considering that C18H12S2IBr is more active at lower concentrations and has comparable toxic effects to fluconazole in rats, this new compound shows some promise in the treatment of fungal infections. Future, more detailed animal studies are needed, that will include drug interactions and molecular toxicity pathways. If the results are promising, clinical studies should follow. Triazolni antifungalni lijek flukonazol danas je najrašireniji antimikotik u svijetu, mahom zato što dobro prodire u tjelesne tekućine i tkiva. Primijećeno je međutim da ulazi u interakciju s drugim lijekovima, a zbog česte uporabe sve je veći i rizik od stvaranja rezistencije na njega. Stoga se traži njegova dostojna zamjena. Godine 2006. sintetiziran je novi derivat tialobenzena - 2,4-ditiofenoksi-1-jodo-4-bromobenzen (C18H12S2IBr). Iako je ugljikova osnovica tog derivata slična onoj flukonazolu, u prvim in vitro istraživanjima pokazao se puno djelotvorniji. Uslijedilo je in vitro istraživanje njegove citotoksičnosti, u kojem se novi spoj pokazao obećavajućom zamjenom za flukonazol. Cilj je ovog istraživanja bio otići korak dalje i istražiti toksičnost C18H12S2IBr in vivo. Odlučili smo se za četverotjedno istraživanje na štakorima Wistar, kojima se je novi antimikotik davao na usta u dozama dva i pol i pet puta nižima od uobičajene doze flukonazola. U životinja nisu primijećene promjene u konzumiranju hrane i vode, ali je relativna težina organa u ženki bila drugačija u odnosu na kontrolnu skupinu, što upućuje na razlike između spolova u metabolizmu lijeka. Za razliku od C18H12S2IBr, flukonazol je prouzročio značajno povišenje razine leukocita i smanjenje neutrofila. Ostale hematološke promjene u odnosu na kontrolnu skupinu bile su slične u skupina koje su primale bilo flukonazol bilo novi lijek. Razlike u imunolokalizaciji citokroma c u jetrima i bubrezima pokazatelj su snažnijeg apoptotičkog učinka novoga lijeka, ali se zbog drugih ključnih pokazatelja (biokemijskih i histopatoloških) ne može ništa zaključiti jer oni ne odgovaraju nalazima citokroma c. S obzirom na to da C18H12S2IBr snažnije djeluje od flukonazola pri nižim koncentracijama, uz podjednake toksične učinke u štakora, ovaj je novi spoj nesumnjivo obećavajuća alternativa liječenju gljivičnih infekcija. Potrebna su daljnja, podrobnija istraživanja u životinja, koja će obuhvatiti interakcije spojeva i molekularne putove toksičnosti. Bude li se lijek i u tim istraživanjima pokazao boljom alternativom, trebalo bi provesti klinička ispitivanja u ljudi.

Vinclozolin exposure throughout pregnancy and its developmental toxicity

It is well known that vinclozolin, commonly used in greenhouse cultivation, causes alterations in the fetal reproductive system. There were no reports about the effects of vinclozolin exposure on the placenta during gestation, therefore, in this study 50 and 100 mg kg−1 body weight per day (bw d−1) doses of vinclozolin were administered by oral gavage to pregnant rats during gestation. They were then subjected to a caesarean section on gestation day 20. Maternal and fetal liver, kidney, heart, thymus, brain, and placenta were examined histopathologically, the placenta and liver tissues were stained immunohistochemically for Vascular Endothelial Growth Factor (VEGF). Morphometric analyses of fetal body lengths, placental measurements, and fetal skeletal staining were performed. The decrease in the placental weight and placental index was significant in the treatment groups. Degenerations in labyrinth region and spongiotrophoblast were detected in the 100 mg kg−1 bw d−1 vinclozolin group. Mononuclear cell infiltration, cellular degeneration and edema in maternal liver samples and an increase in the number of megakaryocytes, cellular degeneration and congestion in fetal liver samples were observed in the treatment groups. VEGF staining was dense in trophoblastic giant cells and spongiotrophoblasts, and less dense in the labyrinth region. VEGF staining increased especially around the central vein in maternal liver. Minor alterations were also observed in the fetal skeleton measurements. These results demonstrated that vinclozolin and/or its metabolites transport to the placenta and induce histopathological changes in the placenta, and maternal and fetal tissues.

Cytotoxic Effects of a Novel Thialo Benzene Derivative 2,4-Dithiophenoxy-1-iodo-4-bromobenzene (C18H12S2IBr) in L929 Cells

The aim of this study was to compare the cytotoxic effects of a newly synthesized thialo benzene derivative 2,4-dithiophenoxy-1-iodo-4-bromobenzene (C18H12S2IBr) and a well-known antifungal agent, fluconazole, in L929 cells. L929 cells were treated with 250, 500, or 1000 µg/mL of C18H12S2IBr and with the same doses of fluconazole. Cytotoxicity tests including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) leakage, and protein content were compared. Glucose and lactate concentrations were measured to determine alterations in metabolic activity. Apoptosis was investigated by TUNEL test and results were supported with survivin enzyme-linked immunosorbent assay. Treatment with C18H12S2IBr resulted in a concentration-dependent cytotoxicity as indicated by MTT, LDH leakage assay, and decreased protein concentration. The loss of cell viability and the increased LDH leakage in 500 µg/mL and 1000 µg/mL C18H12S2IBr and fluconazole groups indicated cell membrane damage and necrotic cell death. In all groups, metabolic activities were altered but apoptosis was not induced. We have previously investigated lower doses of C18H12S2IBr; there was no cytotoxicity in L929 cells. In this study, higher doses caused cytotoxicity and alterations in metabolic activity . When we consider the similar results obtained from fluconazole and especially the lowest dose of C18H12S2IBr, this newly synthesized compound may be a good alternative antifungal agent.

T-2 toxin induces cytotoxicity and disrupts tight junction barrier in SerW3 cells

T-2 toxin, which is produced in grain and grain products as a secondary metabolite by Fusarium species, is also potentially dangerous for human health. Up to date, no study was reported the cytotoxicity of T-2 toxin on SerW3 cells in the perspective of junctional barriers. This study focused on revealing the cytotoxic effects of T-2 on Sertoli cells associated with cell junctional barriers. In the present study, SerW3 cells were exposed to T-2 toxin at 12, 120 and 1200ng/ml doses for 24 and 48h. Cytotoxicity tests including cell viability (MTT), lactate dehydrogenase (LDH) cytotoxicity test and trypan blue exclusion assay were performed. Occludin, ZO-1, N-cadherin and β-catenin were immunolabelled, expressions of occludin and N-cadherin were determined by western blotting. SerW3 cell barrier integrity was measured by transepithelial electrical resistance (TEER). Cytotoxicity caused by T-2 toxin increased in a dose dependent manner, expressions of proteins and TEER measurement decreased. This study may underlie the early targets of T-2 toxin on SerW3 cells mimicking blood-testis barrier in vitro.

Acrolein-mediated neurotoxicity in growing Wistar male rats

Abstract Acrolein is an environmental and food contaminant widely produced by thermal processed organic compounds. It can be used as a biocide and is known to be generated endogenously by lipid peroxidation. This study was designed for evaluating acrolein toxicity in the context of GFAP, MBP, neurofilaments and histological examination of brain tissues and also hematological paremeters in pubertal male rats. 3–4 weeks aged Wistar male rats were exposed to 0.5, 1 and 2 mg/kg/day acrolein for 90 days. According to acrolein exposure results, slight changes observed in hematological parameters, however no significant differences were observed in GFAP and MBP. Unlikely, acrolein caused several histopathological changes in brain tissues as well as neurofilament accumulation. It may be concluded that acrolein might contribute to formation of neurodegenerative disorders.

In vivo toksičnost novoga antimikotika 2,4-ditiofenoksi-1-jodo-4-bromobenzena: proširenje in vitro istraživanja

Abstract Triazole fungicide fluconazole has become the most widely used antifungal agent in the world, mainly because of its ability to penetrate well into body fluids and tissues. However, it has been reported to interact with many drugs and because of its common use, the risk of resistance to fluconazole increases. This calls for new anti-fungal drugs that would be able to replace it. In 2006, a new thialo benzene derivative - 2,4-dithiophenoxy-1-iodo-4-bromo benzene (C18H12S2IBr) - was synthesised with a carbon backbone similar to fluconazole, and, according to the early in vitro tests, much greater efficiency. Followed an in vitro test of its cytotoxicity, in which the new drug showed promising results as an alternative to fluconazole. The aim of this study was take the next step and test C18H12S2IBr toxicity in vivo. We opted for a four-week test on Wistar rats, in which the new antifungal agent was orally applied at doses two and a half and five times lower than those of fluconazole. There were no changes in daily food and water consumption, but weight gain in female rats and relative organ weights changed in the treated groups, pointing to sex-related differences in drug metabolism and effects. Fluconazole significantly increased leukocytes and lowered neutrophils whereas C18H12S2IBr did not, while other haematological changes in respect to the vehicle control were similar between the treated groups. Differences in cytochrome c in the liver and kidney suggested greater apoptotic effect of the new drug, but interpretation remains inconclusive, considering that other key indicators (biochemistry and histopathology) do not support greater toxicity. Considering that C18H12S2IBr is more active at lower concentrations and has comparable toxic effects to fluconazole in rats, this new compound shows some promise in the treatment of fungal infections. Future, more detailed animal studies are needed, that will include drug interactions and molecular toxicity pathways. If the results are promising, clinical studies should follow. Triazolni antifungalni lijek flukonazol danas je najrašireniji antimikotik u svijetu, mahom zato što dobro prodire u tjelesne tekućine i tkiva. Primijećeno je međutim da ulazi u interakciju s drugim lijekovima, a zbog česte uporabe sve je veći i rizik od stvaranja rezistencije na njega. Stoga se traži njegova dostojna zamjena. Godine 2006. sintetiziran je novi derivat tialobenzena - 2,4-ditiofenoksi-1-jodo-4-bromobenzen (C18H12S2IBr). Iako je ugljikova osnovica tog derivata slična onoj flukonazolu, u prvim in vitro istraživanjima pokazao se puno djelotvorniji. Uslijedilo je in vitro istraživanje njegove citotoksičnosti, u kojem se novi spoj pokazao obećavajućom zamjenom za flukonazol. Cilj je ovog istraživanja bio otići korak dalje i istražiti toksičnost C18H12S2IBr in vivo. Odlučili smo se za četverotjedno istraživanje na štakorima Wistar, kojima se je novi antimikotik davao na usta u dozama dva i pol i pet puta nižima od uobičajene doze flukonazola. U životinja nisu primijećene promjene u konzumiranju hrane i vode, ali je relativna težina organa u ženki bila drugačija u odnosu na kontrolnu skupinu, što upućuje na razlike između spolova u metabolizmu lijeka. Za razliku od C18H12S2IBr, flukonazol je prouzročio značajno povišenje razine leukocita i smanjenje neutrofila. Ostale hematološke promjene u odnosu na kontrolnu skupinu bile su slične u skupina koje su primale bilo flukonazol bilo novi lijek. Razlike u imunolokalizaciji citokroma c u jetrima i bubrezima pokazatelj su snažnijeg apoptotičkog učinka novoga lijeka, ali se zbog drugih ključnih pokazatelja (biokemijskih i histopatoloških) ne može ništa zaključiti jer oni ne odgovaraju nalazima citokroma c. S obzirom na to da C18H12S2IBr snažnije djeluje od flukonazola pri nižim koncentracijama, uz podjednake toksične učinke u štakora, ovaj je novi spoj nesumnjivo obećavajuća alternativa liječenju gljivičnih infekcija. Potrebna su daljnja, podrobnija istraživanja u životinja, koja će obuhvatiti interakcije spojeva i molekularne putove toksičnosti. Bude li se lijek i u tim istraživanjima pokazao boljom alternativom, trebalo bi provesti klinička ispitivanja u ljudi.

In vivo toxicity of a new antifungal agent 2,4-dithiophenoxy-1-iodo-4-bromo benzene: a follow up on our in vitro study / In vivo toksičnost novoga antimikotika 2,4-ditiofenoksi-1-jodo-4-bromobenzena: proširenje in vitro istraživanja

Abstract Triazole fungicide fluconazole has become the most widely used antifungal agent in the world, mainly because of its ability to penetrate well into body fluids and tissues. However, it has been reported to interact with many drugs and because of its common use, the risk of resistance to fluconazole increases. This calls for new anti-fungal drugs that would be able to replace it. In 2006, a new thialo benzene derivative - 2,4-dithiophenoxy-1-iodo-4-bromo benzene (C18H12S2IBr) - was synthesised with a carbon backbone similar to fluconazole, and, according to the early in vitro tests, much greater efficiency. Followed an in vitro test of its cytotoxicity, in which the new drug showed promising results as an alternative to fluconazole. The aim of this study was take the next step and test C18H12S2IBr toxicity in vivo. We opted for a four-week test on Wistar rats, in which the new antifungal agent was orally applied at doses two and a half and five times lower than those of fluconazole. There were no changes in daily food and water consumption, but weight gain in female rats and relative organ weights changed in the treated groups, pointing to sex-related differences in drug metabolism and effects. Fluconazole significantly increased leukocytes and lowered neutrophils whereas C18H12S2IBr did not, while other haematological changes in respect to the vehicle control were similar between the treated groups. Differences in cytochrome c in the liver and kidney suggested greater apoptotic effect of the new drug, but interpretation remains inconclusive, considering that other key indicators (biochemistry and histopathology) do not support greater toxicity. Considering that C18H12S2IBr is more active at lower concentrations and has comparable toxic effects to fluconazole in rats, this new compound shows some promise in the treatment of fungal infections. Future, more detailed animal studies are needed, that will include drug interactions and molecular toxicity pathways. If the results are promising, clinical studies should follow. Triazolni antifungalni lijek flukonazol danas je najrašireniji antimikotik u svijetu, mahom zato što dobro prodire u tjelesne tekućine i tkiva. Primijećeno je međutim da ulazi u interakciju s drugim lijekovima, a zbog česte uporabe sve je veći i rizik od stvaranja rezistencije na njega. Stoga se traži njegova dostojna zamjena. Godine 2006. sintetiziran je novi derivat tialobenzena - 2,4-ditiofenoksi-1-jodo-4-bromobenzen (C18H12S2IBr). Iako je ugljikova osnovica tog derivata slična onoj flukonazolu, u prvim in vitro istraživanjima pokazao se puno djelotvorniji. Uslijedilo je in vitro istraživanje njegove citotoksičnosti, u kojem se novi spoj pokazao obećavajućom zamjenom za flukonazol. Cilj je ovog istraživanja bio otići korak dalje i istražiti toksičnost C18H12S2IBr in vivo. Odlučili smo se za četverotjedno istraživanje na štakorima Wistar, kojima se je novi antimikotik davao na usta u dozama dva i pol i pet puta nižima od uobičajene doze flukonazola. U životinja nisu primijećene promjene u konzumiranju hrane i vode, ali je relativna težina organa u ženki bila drugačija u odnosu na kontrolnu skupinu, što upućuje na razlike između spolova u metabolizmu lijeka. Za razliku od C18H12S2IBr, flukonazol je prouzročio značajno povišenje razine leukocita i smanjenje neutrofila. Ostale hematološke promjene u odnosu na kontrolnu skupinu bile su slične u skupina koje su primale bilo flukonazol bilo novi lijek. Razlike u imunolokalizaciji citokroma c u jetrima i bubrezima pokazatelj su snažnijeg apoptotičkog učinka novoga lijeka, ali se zbog drugih ključnih pokazatelja (biokemijskih i histopatoloških) ne može ništa zaključiti jer oni ne odgovaraju nalazima citokroma c. S obzirom na to da C18H12S2IBr snažnije djeluje od flukonazola pri nižim koncentracijama, uz podjednake toksične učinke u štakora, ovaj je novi spoj nesumnjivo obećavajuća alternativa liječenju gljivičnih infekcija. Potrebna su daljnja, podrobnija istraživanja u životinja, koja će obuhvatiti interakcije spojeva i molekularne putove toksičnosti. Bude li se lijek i u tim istraživanjima pokazao boljom alternativom, trebalo bi provesti klinička ispitivanja u ljudi.