Gulnur Take

An investigation on skin wound healing in mice with a taurine-chitosan gel formulation.

Summary. The process of wound healing begins immediately following surface lesions or just after exposure to radiation, chemical agents or extreme temperatures.Taurine (2-aminoethane sulfonic acid), an amino acid containing sulfur, is found in almost all tissues in mammals, playing various important physio-logical roles in each organ. Taurine exhibits an antioxidant effect and is also known to have effects on cell proliferation, inflammation and collagenogenesis. Many antioxidants have been used to eliminate the negative effects of oxygen free radicals on wound healing.The objective of the present study was to examine the wound healing effect in mice of taurine-chitosan gel, which releases taurine slowly over a long time period. Fifty mM of taurine in 1.5% chitosan polymer (TAU-GEL) and 1.5% chitosan polymer (CHI-GEL) were applied to full thickness skin wounds of mice once a day for seven days. After seven days of treatment, lipid peroxide formation-malondialdehyde (MDA) and hydroxyproline (HPX) levels and the tensile strength of wound tissues were measured. All results were compared with those of the untreated control group (CONT). The structural alterations in the skin layers were also histologically investigated.It was found that locally administered TAU-GEL form significantly increased wound tensile strength by decreasing the MDA and increasing HPX levels. These results were supported by histological findings. All observations suggest that taurine gel may be effective in wound healing.

Effects of metformin on mammalian target of rapamycin in a mouse model of endometrial hyperplasia.

OBJECTIVE The effects of metformin on S6K1, which is a crucial effector of mTOR signaling, and on endometrium were studied in a mouse model of endometrial hyperplasia induced by unopposed estradiol or tamoxifen. STUDY DESIGN Forty-eight oophorectomized Balb/c mice were randomly assigned to receive saline, tamoxifen citrate (4 mg/kg), 17-beta estradiol hemihydrate (4 mg/kg), metformin (50 mg/kg), tamoxifen citrate (4 mg/kg) with metformin (50 mg/kg), or estradiol (4 mg/kg) with metformin (50 mg/kg) for 3 days. Histological markers of uterotrophy, including luminal epithelial cell height and density of endometrial glands were quantified for each slide. Immunohistochemical expression of PCNA and S6K1 was evaluated. H-score was used for S6K1 expression. Statistical analysis was performed using Students t-test for comparison of two continous variables and one-way ANOVA for comparison of multiple variables. RESULTS Mice treated either with tamoxifen or estradiol had significantly increased density of endometrial glands and epithelial heights compared to vehicle-only or metformin-only group (p<0.001). Addition of metformin to tamoxifen or estradiol treated mice significantly decreased the density of endometrial glands and epithelial cell heights (p<0.05). Addition of metformin to tamoxifen significantly decreased the H-score of S6K1 (p<0.05) and the immunohistochemical expression of PCNA (p<0.05) in uterine lining epithelium, glandular and stromal cells. Addition of metformin to estradiol significantly decreased the H-score of S6K1 (p<0.05) and the immunohistochemical expression of PCNA (p<0.05) in uterine lining epithelium, glandular and stromal cells. CONCLUSION Metformin seems to have possible antiproliferative effects on the endometrium of estradiol or tamoxifen treated mice via inhibiting the mTOR mediated S6K1 activation.

Effects of Taurine in Cellular Responses to Oxidative Stress in Young and Middle‐Aged Rat Liver

Abstract:  Aging is related with an increased cellular level of lipid peroxides and reactive oxygen species (ROS). The useful effects of taurine as an antioxidant in biological systems have been attributed to its capability to stabilize biomembranes, to scavenge ROS, and to decrease the peroxidation of unsaturated membrane lipids. The aim of the present study was to investigate the effects of taurine on malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), thioredoxin reductase (TR), and endothelial nitric oxide synthase (eNOS) in young and middle‐aged rat liver. There was not a significant difference in liver MDA levels between the control groups of young and middle‐aged rats (P > 0.05). However, liver GSH levels, and GPx and TR activities between the control groups of young and middle‐aged rats were significantly different (P < 0.05). Liver MDA level was significantly lower in the taurine group of middle‐aged rats (P < 0.05). Liver GSH levels, and GPx and TR activities were significantly increased in the taurine group of middle‐aged rats when compared to the control group (P < 0.05). Liver MDA level was significantly lower in the taurine group of young rats than the ones in the control group (P < 0.05). Liver TR activity was significantly increased in the taurine group of young rats when compared to the control group (P < 0.05). Liver GPx activity was not statistically different between the taurine and the control groups in young rats (P > 0.05). Liver GSH levels were not different between the young taurine and the control groups (P > 0.05). Immunohistochemical studies exhibited no change in eNOS activity after taurine injection in young rats. However, in middle‐aged rats, taurine lowered the eNOS reactivity to the same level found in young rats. These results suggested that exogenous taurine might play a role in aging by means of its reducing effects on free radical levels in parallel to an increase in the antioxidant capacity.

Effect of melatonin and time of administration on irradiation-induced damage to rat testes

The effect of ionizing irradiation on testes and the protective effects of melatonin were investigated by immunohistochemical and electron microscopic methods. Eighty-two adult male Wistar rats were divided into 10 groups. The rats in the irradiated groups were exposed to a sublethal irradiation dose of 8 Gy, either to the total body or abdominopelvic region using a 60Co source at a focus of 80 cm away from the skin in the morning or evening together with vehicle (20% ethanol) or melatonin administered 24 h before (10 mg/kg), immediately before (20 mg/kg) and 24 h after irradiation (10 mg/kg), all ip. Caspace-3 immunoreactivity was increased in the irradiated group compared to control (P < 0.05). Melatonin-treated groups showed less apoptosis as indicated by a considerable decrease in caspace-3 immunoreactivity (P < 0.05). Electron microscopic examination showed that all spermatogenic cells, especially primary spermatocytes, displayed prominent degeneration in the groups submitted to total body and abdominopelvic irradiation. However, melatonin administration considerably inhibited these degenerative changes, especially in rats who received abdominopelvic irradiation. Total body and abdominopelvic irradiation induced identical apoptosis and testicular damage. Chronobiological assessment revealed that biologic rhythm does not alter the inductive effect of irradiation. These data indicate that melatonin protects against total body and abdominopelvic irradiation. Melatonin was more effective in the evening abdominopelvic irradiation and melatonin-treated group than in the total body irradiation and melatonin-treated group.

Fallopian damage induced by organophosphate insecticide methyl parathion, and protective effect of vitamins E and C on ultrastructural changes in rats.

The aim of the present study was to examine the effects of subchronic methyl parathion (MP) administration on lipid peroxidation and fallopian tube damage, and to evaluate the preventive effects of the use of vitamins E and C against toxicity. The experimental groups were: rats treated with corn oil (control group), with 5 mg/kg MP and with 5 mg/kg body weight MP plus vitamins E and C (MP + Vit). The groups were given MP by oral gavage for five days a week for four weeks at a daily dose of 5 mg/kg (MP and MP + Vit) using corn oil as a vehicle. Vitamins E and C were injected at doses of 50 mg/kg intramuscularly and 20 mg/kg intraperitoneally, respectively, just after the treatment with MP in the MP + Vit group. The levels of malondialdehyde (MDA) were determined in rat plasma. Electron microscopic ultrastuructural and histopathological changes in the fallopian tissue were examined. MDA levels were higher in the MP group than in the control group, and lower in the MP + Vit group than in the MP group. MP led to deletions in microvilli and marked loss in kinocillia of surface epithelium. But these marked histopathological findings decreased in the MP + Vit group. Multiple doses of MP administration caused some damage in the fallopian tube, and treatment with vitamins E and C after MP could reduce this effect. Toxicology and Industrial Health 2007; 23: 429—438.

Evaluation of caspase-dependent apoptosis during methyl parathion-induced endometrial damage in rats: Ameliorating effect of Vitamins E and C.

The role of reactive oxygen species (ROS) in various diseases of the female reproductive tract has been shown, and oxidative stress is an important component of the mechanism of toxicity of OPIs. Methyl parathion (MPT) is one of the most widely used organophosphate insecticides (OPIs) in agriculture. The aim of the study was to elucidate the effect of subchronic MPT exposure on lipid peroxidation and serum activities of cholinesterase (ChE), and the protective effects of combination of antioxidant Vitamins E and C in rats. Additionally, histopathological and immunohistochemical changes in endometrium were aimed to be examined. Three groups of rats were used in the experiment. The first group was treated with 5mg/kg MPT; the second group was treated with 5mg/kg body weight MPT plus Vitamin E and Vitamin C (MPT+Vit); and the third group was given only corn oil (control). MPT and MPT+Vit groups were given MPT by gavage 5 days a week for 4 weeks at a dose level of 4mg/(kgday) by using corn oil as the vechicle. Vitamins E and C were injected at doses of 50mg/kg i.m. and 20mg/kg body weight i.p. Histopathological and immunohistochemical examinations for caspase-3 and caspase-9 were accomplished in the endometrium. The level of malondialdehyde (MDA) increased significantly in the MPT group compared with the control group (p<0.05). MDA significantly decreased in the MPT+Vit group compared with the MPT group (p<0.05). Administration of Vitamins E and C along with MPT significantly reduced the histopathological changes and the extent of apoptosis. In conclusion, subchronic MPT administration caused endometrial damage and that treatment with a combination of Vitamins E and C reduced endometrial damage caused by MPT.

Continuous or intermittent noise exposure, does it cause vestibular damage?: An experimental study

OBJECTIVE Aim of this study was to compare vestibular changes in guinea pigs exposed to same level of continuous and intermittent noise by electron microscopy. METHODS The study included 10 adult albino guinea pigs. In a silent room, a 4-kHz octave band noise at an intensity of 120 dB SPL was presented. Six animals were exposed to continuous noise for 6h, and four animals were exposed to 12h intermittent noise. One day after noise exposure eight guinea pigs were decapitated and temporal bones of one side were removed. Ten days after continuous noise exposure two guinea pigs were decapitated. They were examined with an electron microscope. RESULTS The most characteristic changes in the macula of the continuous noise exposure group were degeneration of the epithelial cells and separation in their layers. Marked crystolysis and stromal cell apoptosis were also noted in this group compared to the intermittent noise exposure group. Effect of noise was more obvious in the group that continuous noise was applied. The histological changes in group which examined after 10 days were similar to the group that examined after 1 day. CONCLUSION Continuous noise can cause more damage to the vestibular system compared with intermittent noise and histological changes after continuous noise are permanent.

The effect of repeated tryptophan administration on body weight, food intake, brain lipid peroxidation and serotonin immunoreactivity in mice

Tryptophan as a circulating precursor of serotonin (5-HT) may suppress food intake and body weight. Tryptophan administration can enhance the generation of reactive oxygen species (ROS) by inducing oxidative pathway in vivo and in vitro. We have examined the effect of repeated tryptophan administration on food consumption, body weight, brain lipid peroxidation and 5-HT immunoreactivity. Tryptophan was given at the dose of 100 mg/kg/24 hr in 0.2 ml saline solution i.p. for 7 days to mice. Control mice received 0.9% NaCL solution at the same manner and volume. Body weights were recorded at the beginning and end of the experiments. Thiobarbituric acid reactive substance (TBARS), the last product of lipid peroxidation, was measured spectrophotometrically. Brain 5-HT levels were determined by the immunohistochemical method. Our findings indicate that the tryptophan suppresses food intake significantly in mice. Body weight decreased and brain TBARS levels increased significantly by repeated tryptophan treatment. Immunohistochemical detection showed that 5-HT levels increased by tryptophan administration. There is a link between increased 5-HT level and oxidative stress by tryptophan administration on brain tissue. Tryptophan at repeated doses should be exercised carefully in clinical practice.

Retinal ganglion cell toxicity due to oxcarbazepine and valproic acid treatment in rat

PURPOSE To evaluate and to compare the possible toxic effects of oxcarbazepine (OXC) and valproic acid (VPA) on retinal ganglion cells (RGCs) in rat. METHODS Forty female Wistar rats (21-24 days old and weighted between 44.6 and 57.3g) were divided equally into 4 experimental groups which were applied tap water (group 1), 300mg/(kgday) VPA (group 2), 100mg/(kgday) OXC (group 3), and both VPA and OXC (group 4) via gavage for 90 days. Enucleation was performed for histopathologic analysis. RGCs were counted under the light microscopic examination. RESULTS RGC numbers in OXC and combined OXC-VPA groups were found to be lower than those of control group. On the other hand RGC number was comparable with those of control group in VPA group. CONCLUSION OXC seems to be toxic to RGCs at 100mg/kg dose when it is been given as a monotherapy or combined with VPA. Single VPA treatment has no effect on RGC number.

Dose-related immunohistochemical and ultrastructural changes after oral methylphenidate administration in cerebrum and cerebellum of the rat

Methylphenidate is a piperidine derivative and is the drug most often used to treat attention deficit/hyperactivity disorder of children and young adults. Our aim is to investigate dose-dependent dopamine-2 receptor and glial fibrillary acidic protein expression and ultrastructural changes of the rat brain, to demonstrate possible toxicity of the long-term and high dose use of the methylphenidate. In this study, 27 female prepubertal Wistar albino rats, divided into three different dose groups (5, 10 and 20 mg/kg) were treated orally with methylphenidate dissolved in saline solution for 5 days per week during 3 months. At the end of the third month, tissues were removed and sections were collected for immunohistochemical and ultrastructural studies. We believe that methylphenidate causes dose-related activation of the dopaminergic system in several brain regions especially in ventral tegmental area and also causing neuronal degeneration and capillary wall structural changes such as basal membrane thickness and augmentation of the pinostatic vesicle in the endothelial cells. Also, increased dose of Ritalin is inducing astrocytes hypertrophy especially astrogliosis in pia-glial membrane and this is the result of the degenerative changes in prefrontal cortex region due to high dose methylphenidate administration. The dose-related accumulation of the astrocytes in capillary wall might well be a consequence of the need for nutrition of the neuronal tissue, due to transport mechanism deficiency related to neuronal and vascular degeneration. Thus, we believe that the therapeutic dose of methylphenidate must be kept in minimum level to prevent ultrastructural changes.