Folic acid-modified biocompatible Pullulan/poly(acrylic acid) nanogels for targeted delivery to MCF-7 cancer cells.

We prepared a novel nanogel consisting of poly(acrylic acid) (PAA) and pullulan (Pull) via a facile and green irradiation protocol. Synthesized nanogels were modified with bovine serum albumin (BSA) and folic acid (FA) and then loaded with doxorubicin (DOX) to obtain a delivery system with tumor-specific targeting ability and enhanced biocompatibility. In-vitro DOX release was investigated at different pH values, and it was found that DOX release was higher in acidic media, which is an advantage for the internalization of nanoparticles in acidic tumor environment. MTT assay and DAPI staining were performed to evaluate the effects of nanogels on L929 and MCF-7 cells. Based on the results of in vitro studies, DOX-loaded nanogels were found to be effective on cancer cells, while the neat ones were nondestructive in both lines. Overall, we envision that the biocompatible and tumor-specific nanogels could be a promising safe drug carrier system for cancer therapy.

Flusilazole-induced damage to SerW3 cells via cytotoxicity, oxidative stress and lipid metabolism: An in vitro study.

Flusilazole (C16H15F2N3Si) is a triazole fungicide and it is being used widely in recent years to control fungal infections in various fruits and vegetables. This study aims to evaluate the impact of flusilazole on cytotoxicity, ATP-dependent cassette transporter proteins (ABC transporter proteins) in SerW3 cells. In this study, SerW3 cells have administrated with 25, 100, and 200 µM flusilazole, cell viability was performed. The quantity of the cellular lipids was evaluated spectrophotometrically. Moreover, the expression of the ABCA1 and ABCB1 proteins determined by immunofluorescence microscopy. Furtherly, evaluation of the cell death type and measurement of the activity of the antioxidant enzymes was performed. According to the results, flusilazole treatment gave rise to inhibition in cell viability, increase in apoptotic cell number, reduction in cellular lipids, and inhibition in the expression of ABCA1 and ABCB1 proteins. Furthermore, it caused decreases in antioxidant enzyme activities. It may be concluded that flusilazole administration may cause infertility/subfertility. The mechanism of action can be due to cytotoxicity, impairment of the detoxification mechanisms, lipid metabolism, and dysregulation of cell functions.

Flusilazole Induced Cytotoxicity and Inhibition of Neuronal Growth in Differentiated SH-SY5Y Neuroblastoma Cells by All-Trans-Retinoic Acid (Atra)

Objectives: Flusilazole (FLUS) is a broad-spectrum organosilicon triazole fungicide used for protecting economically important cereals and orchard fruits. Considering the exposure route of pesticides, pesticide contamination of food is inevitable. Furthermore, excessive exposure to pesticides causes health problems in both target and non-target organisms. It was aimed to evaluate the effects of the triazole fungicide FLUS on cytotoxicity and neurite extension in differentiated SH-SY5Y neuroblastoma cells. Materials and Methods: The SH-SY5Y cells were differentiated into mature neurons using 10-µM all-trans-retinoic acid (RA) treatment for 7 days. Then the differentiated SH-SY5Y cells were treated with 50, 100 and 200 µM FLUS for 24 h. Afterwards, cell viability assays were performed including crystal violet, neutral red cell viability, and lactate dehydrogenase leakage assays. The morphological examinations were performed and neurite lenghts of the cells were measured in all experimental groups. Results: FLUS treatment induced cytotoxicity in SH-SY5Y cells differentiated with RA. Significant decreases in cell viability percentages were observed. Furthermore, neurite lengths were negatively affected by the treatment of FLUS at the highest concentration. Conclusion: FLUS is a fungicide widely used in agriculture to protect crops from fungal diseases. However, the intensive use of these compounds causes a potential risk to human and environmental health. According to the results of the study, it can be concluded that high concentrations of FLUS cause neurotoxicity by causing neural cell death and adverse effects on neurite outgrowth in differentiated SH-SY5Y cells. FLUS exposure can cause neuronal degeneration in mammals.

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 ß-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.

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

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.

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.

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.

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.