Effect of hydroxychloroquine on oxidative/nitrosative status and angiogenesis in endothelial cells under high glucose condition

Introduction: Under the diabetic condition, sustained production of oxidative/nitrosative stress results in irreversible vascular injuries. A great number of diabetic pathologies, such as inefficient or aberrant neo-angiogenesis emerge following chronic hyperglycemic condition. Lack of enough data exists regarding hydroxychloroquine (HCQ) contribution on angiogenesis during diabetes mellitus. Methods: To better address whether HCQ could blunt or exacerbate oxidative status and angiogenesis under high glucose condition (HCG), human umbilical vein endothelial cells (HUVECs) were exposed to 30 µM HCQ in combination with 30 mM glucose over a course of 72 hours. Viability was measured was evaluated by MTT assay. We used Griess method and TBARS assay to monitor changes in the levels of NO and MDA followed by flow cytometric analysis of ROS using DCFDA. To show the impact of HCQ on cell motility and in vitro angiogenic properties, we exploited routine scratch test and in vitro tubulogenesis, respectively. Results: Our data showed that HCQ diminished cell viability under 5 and 30 mM glucose contents. HCQ significantly decreased the total levels of nitric oxide (NO), malondialdehyde (MDA), and reactive oxygen species (ROS) in both sets of environments. Additionally, inhibitory effects were observed on cell migration after exposure to HCQ (P < 0.001). Anti-angiogenic activity of HCQ was confirmed by the reduction of tube areas under a normal or surplus amount of glucose (P < 0.001). Conclusion: In overall, results suggest that HCQ changes the oxidative/nitrosative status of HUVECs both in 5 and 30 mM conditions. HCQ is able to reduce migration and angiogenic activity of HUVECs irrespective of the glucose content

In vitro assessment of adsorbents aiming to prevent deoxynivalenol and zearalenone mycotoxicoses

The high prevalence of the Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZON) in animal feeds in mild climatic zones of Europe and North America results in considerable economic losses, as these toxins affect health and productivity particularly of pigs from all age groups. The use of mycotoxin adsorbents as feed additives is one of the most prominent approaches to reduce the risk for mycotoxicoses in farm animals, and to minimise carry-over of mycotoxins from contaminated feeds into foods of animal origin. Successful aflatoxin adsorption by means of different substances (phyllosilicate minerals, zeolites, activated charcoal, synthetic resins or yeast cell-wall-derived products) has been demonstrated in vivo and in vitro. However, attempts to adsorb DON and ZON have been less encouraging. Here we describe the adsorption capacity of a variety of potential binders, including compounds that have not been evaluated before, such as humic acids. All compounds were tested at realistic inclusion levels for their capacity to bind ZON and DON, using an in vitro method that resembles the different pH conditions in the gastro-intestinal tract of pigs. Mycotoxin adsorption was assessed by chemical methods and distinct bioassays, using specific markers of toxicity as endpoints of toxicity in cytological assays. Whereas none of the tested substances was able to bind DON in an appreciable percentage, some of the selected smectite clays, humic substances and yeast-wall derived products efficiently adsorbed ZON (>70%). Binding efficiency was indirectly confirmed by the reduction of toxicity in the in vitro bioassays. In conclusion, the presented test protocol allows the rapid screening of potential mycotoxin binders. Like other in vitro assays, the presented protocol combining chemical and biological assays cannot completely simulate the conditions of the gastro-intestinal tract, and hence in vivo experiments remain mandatory to assess the efficacy of mycotoxin binders under practical conditions

Zearalenone and its metabolite exposure directs oestrogen metabolism towards potentially carcinogenic metabolites in human breast cancer MCF-7 cells

Zearalenone (ZEN) is produced by Fusarium species contaminating various agriculture crops. In this study, the effects of ZEN and its metabolites α-zearalenol (α-ZEL), and β-zearalenol (β-ZEL) on the formation of carcinogenic oestrogen-catechols in MCF-7 cells were investigated. To assess the effects of mycoestrogens on the activity of cytochrome P450 1A1 and CYP1B1, the rate of ethoxyresorufin O-deethylation (EROD-assay) was measured. The effects of mycoestrogens on the expression of CYP 1A1, CYP 1B1, aryl-hydrocarbon receptor (AhR), and oestrogen receptor alpha (ERα) were determined by qPCR. The catechol-O-methyltransferase (COMT) activity was measured as the ratio of the methoxy metabolites of oestradiol. Results show that mycoestrogens inhibited significantly the CYP1-dependent EROD activities. In the presence of selective inhibitors, mycoestrogens reduced CYP 1A1 and enhanced CYP 1B1 activity. Quantitative PCR analyses demonstrated the upregulation of AhR and confirmed the selective effect of mycoestrogens on CYP1 expression levels and the decline of the CYP 1A1/CYP 1B1 ratio. Mycoestrogens increased the ratio of 4-MeOE to 2-MeOE2 formation significantly (P < 0.05). Our results suggest that the tested mycoestrogens increase the production of CYP1B1-mediated oestrogen catechol metabolites, directing the biotransformation of E2 towards 4-OHE2, which has been identified earlier as a crucial factor in oestrogen-induced tumour initiation

Silymarin attenuates mycophenolate mofetil-induced duodenal disorders in rats

Objective: The protective effect of silymarin (SMN) on mycophenolate mofetil (MMF)–induced duodenal disorders was investigated. Materials and Methods: Forty-two Wistar rats were assigned to seven groups including control and test groups. The control animals received saline and the test animals were treated with MMF (30 mg/kg, orally) and saline, MMF and SMN (25, 50, and 100 mg/kg, orally), MMF and Celecoxib (CLX, 50 mg/kg, orally), and MMF and SMN plus CLX for 14 consecutive days. The antioxidant status and myeloperoxidase activity were determined and the histopathological examinations on duodenal section also were performed. Results: Biochemical analyses revealed that SMN and CLX individually and in combination therapy could reduce the MMF-increased nitric oxide (NO) content, myeloperoxidase (MPA) activity, and malondialdehyde (MDA) level, while the MMF-reduced level of total thiol molecules (TTM) was increased significantly (

Determination of naturally occurring estrogenic hormones in cow's and river buffalo's meat by HPLC-FLD method

This study was performed to measure and compare the levels of steroid hormones [estrone (E1), 17β-estradiol (E2), and estriol (E3)] and their conjugated metabolites in cow's and river buffalo's meat in two distinct follicular and luteal phases. Moreover, the possible effect of a heating process on steroid hormone concentration was also investigated. The collected meat (biceps femoris muscle) samples were subjected to liquid extraction, enzymatical deconjugation, and C18 solid-phase extraction. Estrogens were analyzed using high performance liquid chromatography equipped with a fluorescence detector. In the follicular phase the levels of steroid hormones (E1 and E2) in either tested species were higher than the luteal phase. Moreover, in the present study, E1 concentration (free and deconjugated value, 16.2 ± 1.1 ng/L) was found to be the highest phenolic estrogen in beef, while the dominant estrogen in muscle of river buffalo was E2 (free and deconjucated value, 23.3 ± 1.3 ng/L). The study revealed that animal species influenced the concentration of hormones (E1 and E2) in the samples. The heating process did not significantly change (p > 0.05) the levels of estrogens. The further findings of the present study showed that E3 (deconjugated form) was only detected in the buffalo's meat (15.8 ± 1.9 ng/L). These data suggest that although meat is one of the valuable nutrient sources for humans, there are, however, increasing concerns about the safety of meat due to the excessive presence of steroid hormones

Crataegus Monogyna Aqueous Extract Ameliorates Cyclophosphamide-Induced Toxicity in Rat Testis: Stereological Evidences

Cyclophosphamide (CP) is extensively used as an antineoplastic agent for the treatment of various cancers, as well as an immunosuppressive agent. However, despite its wide spectrum of clinical uses, CP is known to cause several adverse effects including reproductive toxicity. Crataegus monogyna is one of the oldest pharmaceutical plants that have been shown to be cytoprotective by scavenging free radicals. The present study was conducted to assess whether Crataegus monogyna fruits aqueous extract with anti-oxidant properties, could serve as a protective agent against reproductive toxicity during CP treatment in a rat model. Male Wistar rats were categorized into four groups. Two groups of rats were administered CP at a dose of 5 mg in 5 ml saline/kg/day for 28 days by oral gavages. One of these groups received Crataegus monogyna aqueous extract at a dose of 20 mg/kg/day orally four hours after cyclophosphamide administration. A vehicle treated control group and a Crataegus monogyna control group were also included. The CP-treated group showed significant decreases in the body, testes and epididymides weights as well as many histological alterations. Stereological parameters and spermatogenic activities (Sertoli cell, repopulation and miotic indices) were also significantly decreased by CP treatment. Notably, Crataegus coadministration caused a partial recovery in above-mentined parameters. These findings indicate that Crataegus monogyna may be partially protective against CP-induced testicular toxicity

Chemoprotective effect of Crataegus monogyna aqueous extract against cyclophosphamide-induced reproductive toxicity

AbstractCyclophosphamide (CP) is extensively used as an antineoplastic agent for treatment of various cancers, as well as an immunosuppressive agent. However, despite its wide spectrum of clinical uses, CP is known to cause several adverse effects including reproductive toxicity in humans and experimental animals. Crataegus monogyna is one of the oldest medicinal plant has been shown to be cytoprotective by scavenging free radicals. The present study was conducted to assess whether Crataegus monogyna fruits aqueous extract with anti-oxidant properties could serve as a protective agent against reproductive toxicity during CP treatment in a rat model. Male Wistar rats were categorized into four groups. Two groups of rats were administered CP at a dose of 5 mg in 5 mL saline kg-1 per day for 28 days by oral gavages. One of the groups received Crataegus monogyna aqueous extract at a dose of 20 mg kg-1 per day orally four hours after cyclophosphamide administration. A vehicle-treated control group and a Crataegus monogyna control group were also included. The CP-treated group showed significant decreases in the body and organ weights and spermatogenic activities as well as many histological alterations. CP treatment also caused a significant decrease in sperm count and motility with an increase in dead and abnormal sperms. Moreover, significant decrease in serum levels of testosterone and increased serum concentrations of FSH, LH, LDH, CPK and SGOT were observed in CP-treated rats. Notably, Crataegus coadministration caused a partial recovery in above-mentioned parameters. These findings indicated that Crataegus might be partially protective against CP-induced reproductive toxicity

Assessment of Acute Toxicity and Lethal Dose of Prodigosin in Mice

Background & Objective: Acute toxicity assessment is the first priority in the determination of any related risk to the biologically unknown chemicals to human and animals. LD50 determination as an accepted model of acute toxicity assay in animal models for a new drug in clinical trials is one of the important requirements of the drug launching process. Prodigiosin is a substance extracted from Serratia marcescens and has antitumor and antifungal activities. Thus, in this study, acute toxicity of prodigiosin was determined using the lowest number of laboratory animals. Materials & Methods: In this experimental study, different doses of prodigiosin were administered intraperitoneally in male mice. Twenty four hours after injection, alongside examining behavior of the animals receiving the prodigiosin, some organs including the heart, liver, kidney, spleen, intestine and lung were sampled, and after paraffin block preparation and microtome cutting, they were stained with hematoxylin-eosin and examined by light microscopy. Results: The results of this study indicated that LD50 for prodigiosin is 4500 mg / kg, when administered intraperitoneally and histopathological findings indicate very slight and minor damage to the liver, kidney and spleen, while no remarkable damage on other organs including the heart, lung and intestine was observed. Conclusion: Based on the results of current study and estimated LD50 level, it is suggested that prodigiosin can be categorized as a safe compound with the least histopathological impact on the vital organs

Effect of Aflatoxin B1 on Histomorphometric Parameters of Testis in Male Mature Albino Mouse

Background and Objectives: Aflatoxin B1 (AFB1) is a mycotoxin, which is produced by Aspergillus flavus and Aspergillus parasiticus. Among four types of AFB (AFB1, AFB2, AFB3, and AFN4), AFB1 is the most abundant and toxic in the world. Aflatoxin decreases sperm production and quality. In the present study, the effect of ABF1 was investigated on spermatogenesis, spermiogenesis, and cell apoptosis in the testicular tissue. Methods: In this experimental study, 24 albino mice were divided into 4 groups; control group intraperitoneally received 0.2 ml corn oil and experimental groups received AFB1 (20 &micro;g) for 7, 15, and 35 days, respectively. After 7, 15 and 35 days, the testicular tissue samples were taken, and after preparing tissue sections, hematoxylin and eosin (H&E) staining was performed. Histomorphometric parameters and DNA damage was evaluated using DNA Ladder method. Data analysis was carried out using one-way ANOVA at the significance level of p<0.05. Results: In this study, AFB1 led to tissue damage, especially in germ cells line. The percentage of seminiferous tubules with differentiation indices, tubular repopulation, and negative spermiogenesis, increased in the mice received AFB1; seminiferous tubules diameter and germinal epithelium thickness, significantly decreased (p<0.05). Finally, AFB1 increased the DNA damages. Conclusion: The findings of this study revealed that AFB1 increases testicular cell apoptosis via DNA damages, also it can increase differentiation indices, tubular differentiation, and spermiogenesis in testicular tissue, which consequently lead to decrease in sperm production and quality

A cytotoxicity and comparative antibacterial study on the effect of Zataria multiflora Boiss, Trachyspermum copticum essential oils, and Enrofloxacin on Aeromonas hydrophila

Objective: this study designed to test the antibacterial potency of enrofloxacin (ENR) and essential oils from Zataria multiflora Boiss (ZEO) and Trachyspermum copticum (TEO) on Aeromonas hydrophila. Material and Methods: The antibacterial potency of test compounds was determined by several methods including the inhibition zone diameter determination, microbroth dilution method and colorimetric method of MTT. The cytotoxicity of test substances was assessed on Chinook salmon (Oncorhynchus tshawytscha) embryo (CHSE-214) cells. Results: Results showed that ENR and tested essential oils exert antibacterial effect against A. hydrophila. Moreover, ENR exerted the most potent antibacterial effect with MIC values of 62.5 ng/ml. The natural compounds of ZEO and TEO also showed antibacterial effects with rather high MIC values of 0.315 mg/ml, and 1.25 mg/ml, respectively. None of the tested substances showed toxicity on CHSE-24 cells. Conclusion: It is concluded that ZEO and TEO could be applied to prevent from A. hydrophila infection. Moreover, data also suggest that MTT method could be both cost- and time-effective and accurate method of MIC determination