The effect of glutathione and buserelin on the stereological parameters of the hypothalamus in the cyclophosphamide-treated mice.

INTRODUCTION: New anticancer drugs have increased the survival and fertility rates in young patients. These drugs (i.e., cyclophosphamide; Cyc) have some side effects on the hypothalamus and fertility. One possible chemical for reducing these side effects is thiol or GnRH agonist. This study aimed to evaluate the capability of these agents for reducing the cyclophosphamide effects on the hypothalamus. METHODS: Sixty-three female mice were randomly assigned into seven groups. All groups including the control group had free access to water and mouse chow ad libitum. The sham group received normal saline. The Glu and Bus groups received glutathione (Glu) and buserelin (Bus) daily for 16 days, while the Cyc group received only cyclophosphamide as a single dose; the Cyc + Glu and Cyc + Bus groups, in addition to cyclophosphamide, received glutathione and buserelin, respectively. The volume of the hypothalamus, its neuron number, and dead neurons were evaluated using stereological methods. RESULTS: There was no significant difference in the evaluated stereological parameters between the control and sham groups. However, the animals which received Cyc showed a decrease in the volume of the hypothalamus and its neuron number and density and an increase in cell death as compared with the control group. The treatment of the mice that received Cyc with Glu or Bus prevented these changes. CONCLUSION: This study showed that both GnRH agonist and thiol preserved or improved structural changes in the hypothalamus caused by cyclophosphamide in mice, suggesting that using thiol and especially GnRH agonist along with chemotherapy drugs may have protective effects on fertility.

The influence of interferon-γ on cardiac and renal histopathological changes induced by carbamazepine.

BACKGROUND: Carbamazepine (CBZ) is used for the treatment of epileptic seizures. This study was designed to evaluate the effect of Interferon-gamma on the fetal heart and kidney histopathological changes of CBZ-treated pregnant mice. METHODS: Twenty pregnant mice were divided into four groups. The control group received distilled water. The second group received 240 mg/kg of CBZ by gastric gavage. The third group received intraperitoneal injection (IP) of IFN-γ. The fourth group received IP injection of IFN-γ with 240 mg/kg CBZ by gavage. The fetuses were delivered by hysterectomy on the 18th day of gestation. RESULTS: The mean weight, crown-rump length, the total volume of the heart and kidney of the fetuses in the CBZ-treated group were significantly reduced when compared with the control, INF-γ and CBZ + INF-γ groups (p < 0.05). INF-γ prevented histopathological changes when used with CBZ (p < 0.05). CONCLUSION: CBZ induced structural changes in the fetal tissues of the pregnant mice. However, IFN-γ could reduce these changes (Tab. 1, Fig. 4, Ref. 26).

Improved BALB/c mice granulosa cell functions using purified alginate scaffold.

Alginate, a non-toxic polysaccharide isolated from brown algae, is a widely used 3-dimensional (3D) porous scaffold for the granulosa cell and follicle encapsulation. However, impurities in commercial alginate can lead to alginate biocompatibility reduction. The aim of this study was to evaluate in vitro behavior of the granulosa cells seeded on the purified alginate in varying concentrations compared with matched non-purified ones. We produced a purified alginate using a simple and efficient method. Then, the granulosa cells from mice were isolated and seeded in various concentrations of (0.5%, 1% weight/volume) purified and non-purified alginate. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used on the 3rd, 5th and the 8th days of culture as an index of cell viability and proliferation. Furthermore, the secreted estradiol, progesterone and alkaline phosphatase enzyme (ALP) were measured in the granulosa cells culture media using radioimmunoassay kits. The cells cultured on purified and low concentration alginate showed a higher proliferation rate, sex hormone production and ALP activity. The results confirmed the impact of the alginate hydrogel properties on proliferative rate and function of granulosa cells in a 3D culture system.

Protective Effect of Vitrified-warmed Media with Clove Bud (Syzygium aromaticum) Extract on Mouse Oocytes and Resultant Blastocysts.

BACKGROUND: Although oocyte vitrification has become an essential part of infertility treatments, the formation of reactive oxygen species can reduce the quality of oocytes. OBJECTIVE: Protective effects of vitamin E or clove bud extract in the vitrification and warming media of vitrified mature oocytes were evaluated on blastocysts derived from the warmed oocytes. MATERIALS AND METHODS: Oocytes were vitrified-warmed with vitamin E (0, 50, 100, 200 and 400 mM) or hydroethanolic extract of clove bud (0, 5, 10, 20 and 40 µg/ml) in vitrification and warming media, and resultant blastocysts were evaluated. RESULTS: Mid concentrations of these antioxidants (10 and 20 µg/ml of clove bud extract and 100 and 200 µM of vitamin E) improved the blastocyst formation (P<0.05). The expression of catalase (P<0.01) and superoxide dismutase-1 (Sod1) genes, as well as the inner cell mass number (P<0.05), were significantly less in the blastocyst of the control (untreated) vitrified oocytes, however these levels were restored to normal by clove bud extract. In both antioxidant groups Bcl2l1 gene expression was promoted in comparison to the controls (P<0.05). CONCLUSION: Clove bud extract, with antioxidant and anti-apoptotic properties, may serve to prevent or reduce oxidative stress condition of oocyte vitrification.

Neuroprotective effects of NMDA and group I metabotropic glutamate receptor antagonists against neurodegeneration induced by homocysteine in rat hippocampus: in vivo study.

Homocysteine (Hcy), a neurotoxic amino acid, is a risk factor for neurodegenerative diseases. Previous in vitro studies have demonstrated that group I metabotropic glutamate receptors along with N-methyl-D-aspartic acid (NMDA) receptors participate in acute and chronic aspects of Hcy-induced neuronal damage. In the present study, we examined whether the same mechanism may be involved in homocysteine neurotoxicity in vivo. Memantine, MPEP, and LY367385 were used as NMDA, mGlu5, and mGlu1 antagonists, respectively. Repeated i.c.v injection of Hcy was performed for three consecutive days. Neuronal loss in different zones of the hippocampus was assessed by Nissl, Fluoro-Jade B, and TUNEL staining. Neuronal degeneration was observed in both types of apoptosis and necrosis. All glutamate receptor antagonists, even when given alone, provided some degree of neuroprotection. The degree of protection was dependent on the area of the hippocampus. While memantine was more potent against Hcy-induced apoptosis, the potency of mGluR antagonists in neuronal protection against apoptosis and necrosis was almost equal. No more protection was observed when all three antagonists were used simultaneously. It seems that Fluoro-Jade could be a useful marker of apoptotic cell death. Taken together, results demonstrate that, in vivo, Hcy neurotoxicity is mediated mainly by the NMDA receptors and group I mGluRs.

The effect of the follicular fluid on sperm chromatin quality in comparison with conventional media.

BACKGROUND: Follicular fluid (FF) is biological fluid rich in nutrients, growth factors, hormones and may affect the sperm quality. Sperm washing has been done using conventional media in laboratory procedure so far. AIM: This study aimed to investigate the effects of FF on survival and maintenance of chromatin integrity post swim up. MATERIALS AND METHODS: Each washed semen sample was divided into two parts; the control group was incubated in the media, and the experimental groups incubated in the media containing 10% follicular fluid. Smears were prepared after 20 min, 180 min, 24 hours and no incubation times. Sperm chromatin changes like protamine, histone, DNA denaturation, sperm chromatin stability and motility were evaluated at different times. RESULTS: Incubation of sperm in the follicular fluid increased sperms with normal histone, normal chromatin protamine and sperm with normal head size (p < 0.05). CONCLUSIONS: Administration of follicular fluid into the culture media of the sperms that had been separated by swim up method could improve the sperm quality. Further studies are recommended for understanding the mechanism of the structural change of the sperm chromatin.  

In vitro effects of α-tocopherol on teratozoospermic semen samples.

A strong positive correlation exists between teratozoospermia and reactive oxygen species production, which in turn has negative effects on their in vitro fertilisation outcome. Our aim of this study was to determine potential protective effects of α-tocopherol on teratozoospermia motility, viability, acrosome reaction and DNA integrity after 1-h in vitro incubation. Teratozoospermic semen samples were obtained from 15 volunteers aged between 20 and 30 years after 3-5 days of sexual abstinence. Samples were washed, centrifuged and incubated in 37 °C and 5% CO(2) until sperm swimmed-up. Spermatozoa were counted in the supernatant and divided into four groups, each contained 2 × 10(6) sperm/ml(-1). Groups one to four were incubated for 1 h with Ham's F-10 solution as control group, 10 µm A23187, 40 µmα-tocopherol and 10 µm A23187 + 40 µmα-tocopherol respectively. The results indicated that α-tocopherol has ability to enhance teratozoospermia viability and motility, while there were no ameliorative effects on acrosome reaction and DNA fragmentation. A23187 induced acrosome reaction in teratozoospermia and α-tocopherol significantly diminished this effect. In conclusion, although α-tocopherol could improve teratozoospermia motility and viability, its effects on DNA integrity and acrosome reaction ability as supplementation IVF culture media are not obvious.