Intra-arterial transplantation of neural stem cells improve functional recovery after transient ischemic stroke in adult rats.

OBJECTIVES: The neural stem cell transplantation has been proposed as alternative therapy to promote functional recovery after various neurological disorders. The aim of this study was to evaluate the effect of intra-arterial transplantation of adult neural stem cells on improving local brain ischemia injuries. MATERIALS AND METHODS: In this study, 32 male Wistar rats were used. Ischemia was induced using a middle cerebral artery obstruction with monofilament nylon suture. Neural stem cells were isolated from subventricular zone of the rat brain. 24 hours after local ischemia, the cells were labeled with DiI and transplanted intra-articularly. Evaluation of neurological movement deficits was performed using a neurological deficit score. The transplanted neural stem cells differentiation into neurons and astrocytes was assessed by immunohistochemistry. RESULTS: The results indicated that lesion volumes in the ischemic control, PBS and the treatment groups were 31.5, 29.8 and 14.7 % respectively. Our results also showed that the number of eosinophilic neurons and also neurological impairment in the treatment group was significantly reduced compared to the control and PBS groups. CONCLUSION: The results suggested that intra-arterial transplantation of neural stem cells 24 hours after ischemia, led to a decrease in the volume of brain ischemic lesion and improved neurological outcomes (Fig. 7, Ref. 23).

Protective effects of boron and vitamin E on ethylene glycol-induced renal crystal calcium deposition in rat.

OBJECTIVES: Kidney stone disease is a common form of renal disease. Antioxidants, such as vitamin E (Vit E) and boron, are substances that reduce the damage caused by oxidation. METHODS: Adult male rats were divided into 5 groups (n=6). In group 1, rats received standard food and water for 28 days (control group); in group 2, standard rodent food and water with 0.75% ethylene glycol/d (dissolved in drinking water) (EG Group); in group 3, similar to group 2, with 3 mg of boron/d (dissolved in water) (EG+B Group); in group 4, similar to group 2, with 200 IU of vitamin E injected intraperitoneally on the first day and the 14th day, (EG+Vit E Group); in group 5, mix of groups 3 and 4, respectively (EG+B+Vit E Group). RESULTS: Kidney sections showed that crystals in the EG group increased significantly in comparison with the control group. Crystal calcium deposition score in groups of EG+B (160), EG+Vit E, and EG+B+Vit E showed a significant decrease compared to EG group. Measurement of the renal tubules area and renal tubular epithelial histological score showed the highest significant dilation in the EG group. Tubular dilation in the EG+B+Vit E group decreased compared to the EG+B and EG+Vit E groups. CONCLUSIONS: Efficient effect of boron and Vit E supplements, separately and in combination, has a complimentary effect in protection against the formation of kidney stones, probably by decreasing oxidative stress.

Efficacy of Crocus sativus L. on reduction of cadmium-induced toxicity on spermatogenesis in adult rats.

Cadmium is a toxic heavy metal element, which probably cause infertility by impairment in spermatogenesis. The present work aimed (i) to study the toxic effect of cadmium on spermatogenesis in rat, as well as (ii) the protective effect of Crocus sativus L. on cadmium-intoxicated rats. Cadmium chloride was administered intraperitoneally during 16 days at intervals of 48 h between subsequent treatments. Crocus sativus L. was pre-treated in both of control and cadmium-injected rats. Animals were sacrificed on day 17 after the first treatment. The left cauda epididymis was removed and immediately immersed into Hank's balanced salt solution for the evaluation of sperm count and viability, and left testis was fixed in 10% formalin for histological evaluation. Following contamination with cadmium, a decrease was observed in the number and viability of cauda epididymis sperm, which were increased by Crocus sativus L. pre-treatment (P < 0.05). In addition, cadmium decreased both cell proliferation and Johnsen Scores in the seminiferous tubules, which were reversed by Crocus sativus pre-treatment (P < 0.05). Furthermore, cadmium-induced decrease in the amount of free serum testosterone as well as an increase in lipid peroxidation activity in the testicular tissue was reversed by Crocus sativus L. (P < 0.05). These findings may support the concept that Crocus sativus L. can improve the cadmium toxicity on spermatogenesis.

Derivation of motor neuron-like cells from neonatal mouse testis in a simple culture condition.

Embryonic stem cell (ESC) therapy is an exciting way to treat neurodegenerative disease and central nervous system injury. However, many ethical and immunological problems surround the use of embryonic stem cells. Finding an alternative source of stem cells is therefore pertinent. In this study, spermatogonia stem cells (SSCs) were used to generate mature motor neurons. SSCs were extracted from neonatal testes and cultured in DMED/F12 medium for 3 weeks. Characterisation of SSC-derived ESC-like cells was confirmed by RT-qPCR, immunostaining, alkaline phosphatase activity and their ability to form embryoid bodies (EBs). The EBs were induced by retinoic acid and Sonic hedgehog and trypsinised to obtain single induced cells. The single cells were cultured in neural medium for 18 days. Characterisation of neural precursors and motor neuron-like cells was confirmed by RT-qPCR and immunocytochemical analysis at the 7th day (early stage) and 18th day (late stage), respectively, of culturing. The neural precursors were found to be positive for nestin and Sox2, and a small fraction of cells expressed ß-tubulin III. Upon further differentiation, multipolar neurons were detected that expressed ß-tubulin III and MAP2 markers. Moreover, the expression levels of Olig2 and PAX6 were significantly lower, while HB9, Isl1 and Isl2 expression levels were higher at the late stage when compared to the early stage. These results show that SSCs have the potential to differentiate to motor neuron-like cells and express markers specific for mature motor neurons. However, the functional ability of these cells remains to be evaluated in future studies.

Ferula gummosa Boiss flower and leaf extracts inhibit angiogenesis in vitro.

BACKGROUND: Angiogenesis is a vital process in development as well as in tumor metastasis. Therefore, inhibition of tumor angiogenesis may be an approach for cancer therapy. In this study, we evaluated the effect of Ferula gummosa Boiss flower and leaf extracts on angiogenesis. MATERIALS AND METHODS: Cell growth and cytotoxic effects of different concentrations (0-70 µg/mL) of F. gummosa Boiss flower and leaf extracts were evaluated on the growth of human umbilical vein endothelial cells (HUVECs) using Neutral Red assay. Then, wound healing, in vitro angiogenesis assay and quantitative VEGF gene expression analysis were conducted with the noncytotoxic concentrations of the ethanol extract. RESULTS: Our results indicated that observed HUVECs viability was higher than 60% for both extracts after 24 hours treatment at concentration of 30 µg/mL or lower, whereas cytotoxic effects were observed at higher concentrations or after 48 hours treatment. F. gummosa Boiss flower and leaf extracts inhibited migration and angiogenesis capacity in a concentration-dependent manner (10-30 µg/mL), and down regulated VEGF transcription (20 µg/mL for flower extract and 30 µg/mL for leaf extract). CONCLUSIONS: Our findings revealed that F. gummosa Boiss flower and leaf extracts may contain antiangiogenic compounds, which could be used in preparation of new therapeutic agents for inhibition of tumor angiogenesis. To the best of our knowledge, this is the first report of antiangiogenic effects of F. gummosa Boiss flower and leaf extracts and more studies are needed to identify the effective components of the extracts.

Acute effects of sulphur mustard gas on the number of lymphocytes in the rat's spleen / Efectos agudos del gas mostaza sobre el número de linfocitos en el bazo de rata

Sulphur mustard (SM), commonly known as mustard gas is an alkylating agent that causes serious blisters upon contact with human skin. SM is frequently used as a chemical warfare agent. There is some evidence for sulfur mustard-induced lymph system effects in humans. Between 2000-2001, 42 male albino Wistar rats were used. After accommodation with environment, we divided rats to control, sham and experimental groups (2.5 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg and 40 mg/kg). Then we injected sulphur mustard oil in rat's intraperitoneal space. Then their spleens were removed for histological verification. Our results showed that significant difference in lymphocytes number in experimental groups after 24 hours. The number of lymphocytes in 5, 10, 20 and 40 mg/kg groups was increased and this increase in 40 mg/kg group was more than the other groups. We concluded that the number of lymphocytes increased due to exposure of mustard gas and there is a relationship between the increase of lymphocytes and dose of exposure.

El sulfuro de mostaza (SM), comúnmente conocido como gas mostaza, es un agente alquilante que causa graves ampollas en contacto con la piel humana. SM se utiliza con frecuencia como un agente de guerra química. Hay algunas evidencias que indican que el SM induce efectos en el sistema linfático en seres humanos. Entre los años 2000-2001, fueron utilizadas 42 ratas albinas Wistar macho. Después de la acomodación con el medio ambiente, las ratas se dividieron en grupos control, impostor y experimental (2,5 mg / kg, 5 mg / kg, 10 mg / kg, 20 mg / kg y 40 mg / kg). Luego se inyectó aceite de SM en el espacio intraperitoneal de las ratas. A continuación, sus bazos fueron removidos para la verificación histológica. Los resultados mostraron una diferencia significativa en el número de linfocitos en el grupo experimental después de 24 horas. El número de linfocitos en los grupos de 5, 10, 20 y 40 mg / kg fue mayor siendo este incremento en el grupo de 40 mg / kg más alto que en los otros grupos. Concluimos que el número de linfocitos aumenta debido a la exposición de gas mostaza existiendo una relación entre el aumento de linfocitos y la dosis de exposición.