[Effect of lavender ethanolic extract on infarct volume in rats subjected to ischemia-reperfusion]

Background: lavender belongs to the Labiatae family and possesses antioxidant acivity

Objective: the present study was conducted to evaluate the protective effect of lavender extract on infarct volume and its possible mechanisms in an experimental model of stroke

Methods: lavender extract [100, 200 mg/kg body weight, ip] was injected for 20 consecutive days. Two hours after the last dose, cerebral artery ligation surgery was performed and 24 hours after induction of ischemia, infarct volume was assessed. Also the amount of serum nitric oxide [NO] level was measured

Results: treatment of rats with lavender extract at a dose of 200 mg for 20 days resulted in a significant decrease in the infarct volume caused by stroke in penumbra area [cortex] and the core [sub-cortical] of brain compared to the control [P=0.044, P=0.047, consecutively]. Lavender extract at a dose of 200 mg significantly increased blood levels of nitric oxide

Conclusion: the results of this study suggest that Lavender extract protects brain against ischemia and reduces infarct volume in rats subjected to ischemia. The mechanism may be related to augmentation in endogenous antioxidant defense in the rat brain. Lavender plant extracts with increasing levels of endothelial nitric oxide, by inhibiting the decrease in cerebral blood flow reduced infarct volume

[Culture of human keratinocytes in serum free medium]

Keratinocytes are useful for cellular transplantation studies in order to improve functional outcome in burn patients and chronic wounds. Recently they have been used for generation of iPS cells with high efficiency. In this study, we described the details on separation, culture and proliferation of human keratinocytes from foreskin samples. In this experimental and qualitative study we obtained neonatal foreskin samples following newborn circumcision under sterile conditions. We used dispase enzyme to separate the epidermis from the dermis. Trypsin enzyme was used for isolation of keratinocyte cells from the epidermis layer. Isolated cells were cultured in type I human collagen-coated dishes and serum-free Epilife medium. We assessed morphological and immunocytochemical aspects of the isolated cells. Morphological and Immunocytochemical analyses revealed that isolated cells have typical keratinocyte morphology and they could express CK14 which is a specific marker of the keratinocytes. The cells were successfully sub-cultured at a split ratio of 1:2 every 4 to 5 days. After passage 10, a significant decrease in the proliferation of the human keratinocyte cells was observed. Morphologically the cells were flat and thin. In this study we improved the method of isolation and cultivation of human keratinocyte from foreskin. Using this method, we isolated human keratinocytes for more than 20 times. Thus, it can be concluded that this method is reproducible in other laboratories

[Generation of induced pluripotent stem cells from mesenchymal stem cells of human umbilical cord vein]

The aim of this study was to generate induced pluripotent stem cells from human umbilical cord vein mesenchymal stem cells by use of plasmid vector. In this experimental study using type IV collagenase enzyme, we extracted and cultured mesenchymal stem cells obtained from human umbilical cord vein wall.Usingelectroporation method, these cells were transfected with plasmid vector which carried selfrenewal transcription factors of OCT4 and SOX2.After 9 days we observed the induced pluripotent stem cells like colonies. The nature of thesecells were evaluated byimmunocytochemistry and alkalin phosphatase activity of embryonic stem cells. Immunocytochemical analysis showed that these cells express the pluripotency markerssuch as OCT4, SSEA4, TRA1-60, TRA1-81, and also hadalkalin Phosphatase activity. This study revealed that transient expression of self renewal genes of OCT4 and SOX2 could lead to development of induced pluripotent stem cells like colonies from umbilical cord vein mesenchymal stem cells

[Isolation of rat embryonic cortical neural stem cells and induction of differentiation]

Neural stem cells [NSCs] play an essential role in the development of embryonic nervous system and have the capacity for self-renewal in the adulthood which may be important for normal functions of the CNS, such as learning, memory and response to injury. These cells exist at different stages of development in different areas of the CNS. The purpose of this study was isolation and confirmation of stem cell and induction of differentiation of the cells isolated from the 15 day old embryonic rat cortex. This was an experimental study on 15 day old embryonic rat cortices [n=6]. 15 days after plug formation in female rats, the animals were transported to the laboratory. Under anesthesia and sterile condition, embryos were removed from the uterus. The meninges were separated by use of a stereoscope and the cortices of the embryos were dissected in HBSS buffer. Then the cortices were cut into small pieces and cultured in DMEM-F12 medium containing bFGF, EGF and B-27 supplement. The medium changed every day to keep the cells in an undifferentiated and proliferative state. DFN medium [DMEM-F12 and supplements N2] without growth factors was used for induction of differentiation. Immunocytochemical technique was used for confirmation of stem cells and detection of various neural cell types. Immunocytochemical evaluation revealed that, these cells were neural stem cells [nestin positive] and had the potential to differentiate in to the neuronal [expression of Beta tubulin III], oligodendrocyte [expression of OSP marker] or astrocyte [expression of GFAP marker]. This is a reliable method for isolation of embryonic neural stem cells and considering their embryonic origin; they can be used to investigate the effect of various agents on the process of CNS development. Also they might be effective in the treatment of neurodegenerative lesions

Evaluation of the expression of self renewal genes [Oct-4, Nanog, Sox2, Nucleostemin, Bmi and Zfx] in bladder, colon and prostate cancers and in cancer cell lines [LNCaP, HepG2, HT-29, CaCo[2], HT-1376]

Uncontrolled self renewal plays a direct function in different types of carcinoma progression. Here we examined the expression of self renewal regulatory factors such as Oct4, Nanog, Sox2, Nucleostemin, Zfx, Bmi-1 in colon, prostate, bladder and liver cancers in human samples and cancer cell lines. We used RT-PCR to examine the expression of these genes in 10 tumors of bladder, 5 tumors of prostate, 5 tumors of colon, 5 normal tissues of colon, and cancer cell lines. The expression of Oct-4 and Nucleostemin at protein level was further determined by immunocytochemical [ICC] analysis in cancer cell lines. We designed specific primers to amplify a segment of Oct4, Nanog, Sox2, Nucleostemin, Bmi and Zfx. As expected DNA fragment of these genes based on designated primer was amplified in the PCR reaction. We detected the expression of these genes in almost all of the examined tumor samples and cancer cell lines that we used. Oct4 and Nucleostemin proteins were expressed in both nuclear and cytoplasmic in cancer cell lines. No immunoreactivity was observed in negative controls, which were incubated in the absence of primary antibody. Collectively, our results indicated that in a tumor population a rare subpopulation of cells within the tumor cell mass has the potential of self renewal, and suggested that their expression can be used as potential tumor markers in diagnosis and/or prognosis of tumors. These results confirm the potential value of the cancer stem-cell theory in cancer therapy

[Assessment of the expression of self renewal genes [Tcl1, Tbx3, Dppa4 and Esrrb] in bladder, liver, colon and prostate cancers]

Here we examined the expression of self renewal regulatory factors such as, Esrrb, Tcl1, Tbx3 and Dppa4 in several tissue samples of cancers and cancer cell lines. These genes are required for efficient self renewal of embryonic stem cells. Caco2, HT-29, HT 1376, Ln Cap, and HepG2 cells were cultured in T25 flasks. Considering the clinical and laboratory findings, human tumor samples were obtained under direct supervision of the medical specialists. Then we evaluated expression of self renewal genes [Tbx3, Tcl1, Esrrb, Dppa4] by reverse transcriptase polymerase chain reaction [RT-PCE] in the above mentioned cells and human tumor samples. To confirm the validity of the laboratory tests, we studied negative control samples and internal control genes. Our data revealed the expression of self renewal genes [TCL1, TBX3, ESRRB and DPPA4] in bladder, liver, prostate and colon cancers and also cancer cell lines. Colon, liver, prostate and bladder cancer cells can express TCL1, TBX3, ESRRB and DPPA4 genes, which are specific markers of stem cells. Therefore in malignant cells of the above mentioned cancers, some cells have the characteristics of stem cells and can play an essential role in the proliferation of malignant cells

[Assessment of functional recovery of contusive spinal cord injury by administration of [-]-Deprenyl in rats]

In this study, the functional recovery of female rats with contusive spinal cord injury [SCI] model was evaluated after administration of [-]-deprenyl. A total of 18 Sprague Dawley female rats were selected for the study and randomly allocated into equal groups [n=6]; control, sham and [-]-deprenyl-treated groups. All animals were laminectomized at T13 level. Based on the weight dropping technique contusion was induced in both control and [-]-deprenyl-treated groups. [-]-deprenyl group received daily injections of 0.1 mg/kg [-]-deprenyl and other group received intra peritoneal [IP] injection of equal amount of normal saline for 14 days. BBB test was carried out in all groups at the first day and at the end of each week after induction of injury for eighth weeks. Spared tissue volume and the number of motoneurons at the site of lesion were measured and compared by means of frozen sections of spinal cord. In contrast to control group, [-]-deprenyl-treated group showed a significant increase in motor ability at all times except for the first day [P <0.05]. In the [-]-deprenyl-treated group the mean volume of spared spinal cord and the mean number of motor neurons were more than those of control group significantly [P<0.05].The results of this study indicated that [-]-deprenyl probably protected motor neurons and spinal cord white matter; hence, it caused motor recovery in contusive SCI model in female rats

[Induction of differentiation of bone marrow mesenchymal stem cells of rat into hepatocyte and study of their biochemical factors]

Bone marrow mesenchymal stem cells [MSCs] are multi potential and capable of differentiating into specialized tissues. The aim of this study was to investigate the induction of differentiation of the bone marrow MSCs of rat into hepatocytes by use of fibroblast growth factor-2 [bFGF], hepatocyte growth factor [HGF] and oncostatin M [OSM] in order to find a suitable source of hepatoecytes for bioartificial liver or liver transplantation. In this research MSCs were obtained from the bone marrow of rat and cultured in DMEM-LG medium supplemented with 15% FBS. These cells were treated with differential medium supplemented with HGF, bFGF and OSM for 24 days. Morphology, RT-PCR and Biochemical assays were used to identify the differentiation of stem cells into hepatic cells. Result: MSCs took a round shape after differentiation, while undifferentiated cells had fibroblast-like morphology. Albumin, urea and alpha-fetoprotein [AFP] were detected in differentiated cells. Alkaline phosphatase activity was confirmed by biochemical tests. The mRNA expression of CK-18 and tyrosine amino transferase [TAT] in differentiated cells was demonstrated by RT-PCR after induction. The rat MSCs from bone marrow can differentiate into hepatocyte-like cells in the presence of HGF, bFGF and OSM in vitro. Cytokines may play an important role in differentiation of bone marrow MSCs of rat into hepatocytes. Bone marrow derived MSCs are a new source of cell types which can be used for cell transplantation in hepatic diseases

[Assessment of capability of fetal and adult neural stem cells to differentiate into endothelial cells]

Neural stem cells exist in various regions of the developing and adult central nervous system [CNS]. They are undifferentiated cells, capable of both self-renewal and producing neurons and glial cells. In addition to generating different types of neural cells, NSCs are capable of producing cells of other tissues. In this study NSCs were isolated from mouse brain and their capability of differentiation into endothelial cells was evaluated. Neural stem cells [NSCs] were isolated from lateral wall of the lateral ventricle of the adult and fetal mouse brain and cultured in serum-free DMEM-F12 medium in the presence of basic fibroblast growth factor [bFGF], epidermal growth factor [EGF] and B27 supplement. Neurospheres were plated on the fibronectin coated culture slides. DMEM-F12 media supplemented with 10% FBS was used for differentiation of NSCs into endothelial cells. Differentiated cells were evaluated by Immuno-histochemistery, RT-PCR and tube formation assays. The results of this study revealed culture of fetal neural stem cells in fibronectin coated and also DMEM-F12 media containing 10% FRS led to differentiation of NSCs into endothelial cells. When differentiated cells were transferred into extra cellular matrix or matrigel, they produced capillary like structures characteristic of endothelial cells. The results of immuno-histo-chemistry and RT-PCR showed differentiated cells can absorb low density lipoproteins and express CD31, VE, cadherin and Flk-1 genes, and also can adhere to BS-1lectin. Unlike neural stem cells isolated from mouse fetus, the isolated cells from adult mouse brain did not differentiate into endothelial cells.This study reveals neural stem cells isolated from fetal mouse brain and adult mouse brain do not show similar behavior in differentiating into endothelial cells

Evaluation of the efficiency of pIRES2 EGFP and pcDNA3-hBDNF-v5 plasmids in transfection of CCE ES cells by the electroporation method

Unlimited self renewal and potential capacity of embryonic stem cells [ESCs] in differentiating into a wide variety of cell types has made the cells an attractive source of donor cells for developmental studies and cell therapy. The aim of the present study was the evaluation of the transfection efficiency of pIRES2-EGFP and pcDNA3-hBDNF-v5 plasmids in CCE ES cells by the electroporation method. The plasmids transformed into DH5

effect of folic acid on plasma zinc concentration in female rats

Considering the relation of teratogenecity with decreased plasma zinc level and expression of hypotheses concerning effect of folic acid on absorption of zinc, this study was designed to deal with the effect of folic acid on plasma zinc concentration in female rats. In this experimental study we assigned 12 albino rats randomly in 2 groups [6 rats in each group]. The rats had been bred in equal environmental and nutritional slates. Control and experimental groups received intra-peritoneal injections of normal saline 4 CC/Kg and folic acid 4 Mg/Kg respectively. All the rats were beheaded and plasma zinc concentration was measured by use of flameless atomic absorption method. Quantitative data presented as Mean +/- SD, were analyzed by means of T-test. Mean plasma zinc levels in the experimental and control groups were 27.90 +/- 2.23, and 42.1 +/- 0,579 respectively [p=0.000] [t=15.14]. These results indicate that intra-peritoneal injection of folic acid decreases plasma zinc concentration in rats. This adverse effect of folic acid may play a role in the development of congenital defects. Therefore we believe that administration of zinc during pregnancy is at least as important as the administration of folic acid, to prevent occurrence of congenial anomalies