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AIM: The aim of this study was to evaluate the effects of preventive vitamin E (α-tocopherol) on antral follicle development and embryogenesis of oocytes obtained after vitrification of mouse ovarian tissue. METHODS: Female Balb/c mice were killed by cervical dislocation after the injection of pregnant mare's serum gonadotrophin (10 IU) and their ovaries were randomly divided into three groups: control or non-vitrified (n = 10), vitrification 1 (5, 10% ethylene glycol + 5, 10% dimethylsulfoxide) (n = 15), and vitrification 2 (10, 15% ethylene glycol + 10, 15% dimethylsulfoxide) (n = 15) with ascending concentration of cryoprotectants. After toxicity tests and vitrification-warming, mechanically isolated antral follicles were cultured in α-minimum essential medium, which was supplemented with or without α-tocopherol (100 µM). The follicular maturation rates and embryo development were collected and assessed. Also, the viability, morphology and ultrastructure of derived antral follicles from vitrified ovaries were analyzed. RESULTS: The morphology and ultrastructure of follicles were well preserved in the vitrified groups and α-tocopherol supplementation of culture media significantly increased the proportion of oocytes that reached metaphase II blastocyst rates compared to non-α-tocopherol supplemented media (P < 0.01). CONCLUSION: Vitamin E improves in vitro maturation rates and blastocyst rates of oocytes that are isolated from vitrified ovarian tissue.
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Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/embriologia , alfa-Tocoferol/farmacologia , Animais , Células Cultivadas , Meios de Cultura , Feminino , Fertilização , Camundongos , Camundongos Endogâmicos BALB C , Oócitos/efeitos dos fármacos , Folículo Ovariano/ultraestrutura , VitrificaçãoRESUMO
The present study was designed to determine changes in spermatogenesis in adult mice after an intraperitoneal injection of vincristine. The effect of a GnRH antagonist synchronous to chemotherapy, which might protect spermatogenesis by halting cell division in spermatogenic cells, was also investigated. Method and Materials: A total of 30 adult male mice were studied in three equal groups of ten. In the V group, a single dose of the chemotherapy drug vincristine was injected intraperitonally at 1.5 mg/kg. In the V+C, group, the injection of Cetrorelix was started, and one week before to one week after vincristine injection continued ( for 3 weeks). Controls received no treatment. Samples were taken from the testicles, and fixed in Boueins fixative for light microscopy. Results: Comparing the mean number of Sertoli and spermatogony cells and the rate of spermatogenesis index (SI) in the V group with controls showed significant differences,which were not evident in the V+C group. Conclusion: According to the results, the cetrorelix antagonist (GnRH) could largely prevent side effects of vincristine administration regarding seminiferous tubules.
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Antineoplásicos Fitogênicos/toxicidade , Hormônio Liberador de Gonadotropina/análogos & derivados , Hormônio Liberador de Gonadotropina/antagonistas & inibidores , Antagonistas de Hormônios/farmacologia , Túbulos Seminíferos/patologia , Espermatogênese/fisiologia , Vincristina/toxicidade , Animais , Células Cultivadas , Hormônio Liberador de Gonadotropina/farmacologia , Masculino , Camundongos , Túbulos Seminíferos/efeitos dos fármacos , Espermatogênese/efeitos dos fármacosRESUMO
The c-Rel transcription factor is a unique member of the NF-kB family that has a role in apoptosis, proliferation and cell survival. Overexpression of c-Rel is detected in many human B cell tumors, including B-cell leukemia and several cancers. The study aimed to investigate the effects of c-Rel siRNA on the proliferation and apoptosis of relapsed pre-B acute leukemia cells. The c-Rel siRNA was transfected into Leukemia cells using an Amaxa cell line Nucleofector kit L (Lonza). Quantitative real-time RT-PCR (qRT-PCR) and western blot were done to measure the expression levels of mRNA and protein, respectively. The flow cytometry was used to analyze the effect of c-Rel siRNA on the apoptosis and proliferation of Leukemia cells. Observed c-Rel expression in the 5 pre-B Acute lymphoblastic leukemia (ALL) patients were higher than the normal cells. The c-Rel siRNA transfection significantly blocked the expression of c-Rel mRNA in a time-dependent manner, leading to a strong growth inhibition and enhanced apoptosis (P<0.05). Our results demonstrated that c-Rel plays a fundamental role in the survival. Therefore, c-Rel can be considered as an attractive target for gene therapy in ALL patients. Also siRNA-mediated silencing of this gene may be a novel strategy in ALL treatment.
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Apoptose/fisiologia , Técnicas de Silenciamento de Genes/métodos , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Proteínas Proto-Oncogênicas c-rel/antagonistas & inibidores , Adolescente , Western Blotting , Proliferação de Células/genética , Células Cultivadas , Criança , Pré-Escolar , Feminino , Citometria de Fluxo , Terapia Genética/métodos , Humanos , Masculino , Proteínas Proto-Oncogênicas c-rel/genética , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , TransfecçãoRESUMO
Purpose: Median septum of Juglans regia L. (Juglandaceae) with anti-diabetic effects has been used in Iranian traditional medicine. The present study estimates both oral acute and subchronic toxicities. Methods: In the oral acute toxicity study, female Wistar rats were treated with doses of 10, 100, 1000, 1600, 2900 and 5000 mg/ kg of the Juglans regia septum of methanol extract (JRSME), and were monitored for 14 days. In subchronic study, JRSME was administered by gavage at dose of 1000 mg/kg daily in Wistar rats for 28 days. Antioxidant status and biochemical examinations were fulfilled, and the vital organs were subjected to pathological analyses. Results: The extract did not produce any toxic signs or deaths; the medium lethal dose must be higher than 5000 mg/kg. In subchronic study, No significant morphological and histopathological changes were observed in the studied tissues. There was a significant increase in serum malondialdehyde (MDA) level in treated group compared to control after 4 weeks of JRSME intake. The treatment of rats resulted in a significant reduction of serum urea level (p<0.05), kidney's xanthine dehydrogenase (XDH) activity (p<0.001) and elevation of aldehyde oxidase (AO) activity (p<0.05) in kidney. In the treated group, the mean diameter of glomerulus and proximal urine tube epithelium stature was slightly greater than control group. A significant increase in serum MDA level is subject for further studies. Conclusion: This study showed that the extract has no acute or subacute adverse effects with dose of 1000 mg/kg. The administration of JRSME may improve kidney structure and function and help in treatment of some chronic diseases.
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BACKGROUND: The aim of the present study was to characterize the morphological and ultrastractural of mouse ovarian tissue with different cryoprotectant solution. OBJECTIVE: Aim of this study, is to demonstrae an improved convetional vitrification method on mouse ovarian tissue using different concentrations of ethylene glycol (EG) and/or dimetyl sulfoxide (DMSO) and EG. MATERIALS AND METHODS: Mouse ovarian tissue dissected and were randomly assigned to three groups: control, conventional vitrification (CV) and toxicity test. Then ovaries were vitrified by 5%, 10% EG or DMSO CV1-CV4, 5%, 10% EG plus DMSO CV5-CV6 and EG plus DMSO in climbing concentrations CV7. The effect of cryoprotectant solutions on ovarian tissue were evaluated by histological examination hematotoxillin & eosin stain, H&E, viability assessment trypan blue stain and ultrastructural analyses transmission electron microscopy, TEM. RESULTS: Ovarian tissue frozen in CV7 solution showed a higher percentage of morphologically normal follicles or viable follicles than other cryoprotectant solutions P < 0.05. Ultrastructural analysis of ovarian tissue showed that less damage was observed in CV7 and it was very similar to the control group. CONCLUSION: Vitrification of ovarian tissue with optimal cryoprotectant solutions such as EG plus DMSO is the most effective for preserving the structural efficiency of ovarian follicles.
Assuntos
Criopreservação/métodos , Folículo Ovariano/ultraestrutura , Animais , Crioprotetores/farmacologia , Feminino , Camundongos , Microscopia Eletrônica de Transmissão , VitrificaçãoRESUMO
PURPOSE: Anti cancer drugs is one of the most important chemotherapeutic factors which can influence spermatogenesis process and germinal epithelium. Since dividing cells are mainly affected by anticancer drugs, the aim of the present study is to investigate the preventive effect of GnRH antagonist on spermatogenic defect produced by anticancer drugs. METHODS: In the present study thirty adult male mice aging 6-8 weeks were divided into 3 groups as: Control, Experimental 1 and Experimental 2. Experimental 1 group received Cisplatin for 5 days as 2.5 mg/kg intraperitoneally and Experimental 2 group received 0.25 mg/kg cetrorelix (GnRH antagonist) one week before cisplatin treatment and continued for 3 weeks. The mice in all groups were sacrificed 35 days after the last injection and testis specimens were fixed in boueins, formaldehyde fixative and 2.5% Glutaraldehide then prepared for light and electron microscopic examination. RESULTS: Light microscopy (LM) study showed that the number of spermatogonial cells, thickness of germinal epithelium, was decreased in Experimental 1group. Electron microscopy revealed that in this group several intercellular spaces appeared between spermatogenic cells and secretory granules in interstitial cells was increased. There were several vacuolated mitochondria and destroyed organelles in spermatogonial cells but in Experimental 2 group condition was similar to control group. CONCLUSION: These results indicate that the cetrorelix administration before cancer treatment may protect germinal epithelium against side effects of cisplatin.
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Ovarian tissue freezing or cryopreservation might be the only acceptable method for preserving the young women fertility, before radiotherapy or chemotherapy. This technology might be used for patients with recurrent ovarian cysts or endometriosis, without ovarian stimulation. Many efforts have made to improve cryopreservation conditions that should be seriously considered for cancer patients. Vitrification is a process which prevents ovarian tissue from cryo damage, then preserves cell viability. Both methods have used for evaluating not only the follicular development, but also the fertility after freezing and thawing. In this manuscript, we have discussed the techniques of ovarian tissue vitrification, then graft and maturation or follicular development is also mentioned.
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PURPOSE: The present study was designed to explore the effect of intraperitoneal administration of cisplatin in germinal epithelium of mice. There are few reports on the side effect of cisplatin on spermatogenesis when are used as anticancer drug. Therefore, in the present study the effect of cisplatin on spermatogenesis was evaluated by electron microscopy. METHODS: Twenty balb/c mice aging 6-8 weeks was used in this study. The mice were divided into two groups, control and cysplatin treated. cysplatin was injected for five days as 2.5 mg /kg. The mice were sacrificed after 5 weeks and testicular specimens were removed, fixed in boueins, formaldeyd fixative and 2.5% Glutaraldehide then prepared for light and electron microscopic study. RESULTS: Observation with optic microscope in treated group thickness of germinal epithelium was reduced a lot and increased the number of apoptotic cells. In some seminiferous tubules only sertoli cells were observed and nucleus of spermatogony cells was hetrochromatin. The electron microscopic observations showed some irregularity waviness and thickening in basal layer. Also myoid cells of this group were thick and contracted. In this group many apoptotic cells and damaged organelles were seen. CONCLUSION: It was indicated that cisplatin affected testicular germinal epithelium by both cytotoxic effect and induction of apoptosis.