Effect of human chorionic gonadotropin and vitamine E on cellular density of CA1 hippocampal area, learning ability and memory, following ischemia - reperfusion injury in mice

Reduction in cerebral blood flow following cereblal ischemia cause the production of oxygen free radicals and finally leads to brain tissue destruction. Pyramidal cells of the CA1 region of hippocampus are highly sensitive to hypoxic condition. This study was done to determine the effect of human chorionic gonadotropin [hCG] and vitamine E on cellular density of CA1 hippocampal area, learning ability and memory, following ischemia - reperfusion injury in mice. This experimental study was done on 40 male mice in 5 groups as follow: sham control, ischemia, hCG treated, vitamine E treated and hCG + vitamine E treated groups. Single dose of vitamin E was injected intraperitonaly during the establishment of reperfusion and hCG was injected from 48h after ischemia for 5 days. Folowing the treatment period, mice brains were fixated by transcardial perfusion and stained by nissle method. The shuttle box was used to evaluate the learning memory. Co-administartion of vitamine E and hCG, significantly increased the cell numbers in hippocampus compared to the ischemic group [P<0.001]. Also learning and memory improved in treatment group in comparison with ischemia group [P<0.05]. Co-administration of vitamin E and hCG improved ischemia-induced neurodegenration and memory impairment

Protective effect of Adenosine A1 receptor and ascorbic acid on hippocampal neuronal density and memory disorder in ischemia reperfusion induced Rats

Brain ischemia is one of the most important factor of morbidity and mortality and leaving many people with mental and physical disabilities. Until now there are no appropriate medications to prevent and cure ischemic injury. This study was done to evaluate the protective effect of Adenosine A1 receptor and ascorbic acid on hippocampal neuronal density and memory disorder in ischemia reperfusion induced Rats. This experimental study was performed on the hippocampus pyramidal neurons on 56 male BALB/c mice. Animals randmly allocated into 8 groups [N=7] including: 1] intact, 2] ischemic control group, 3] ischemic, plus agonist and adenosine of A1 receptor, 4] ascorbic acid [100 mg/daily], 5] ischemic plus agonist adenosine receptor [1 mg/1 kg] one week after ischemia, 6] ischemia, ascorbic acid befor and after ischemia and A1 receptor [1 mg/1 kg] agonist after ischemia, 7] A1 receptor, antagonist [2.25 / 1 kg], one weed after ischemia, 8] Ascorbic acid [100 mg/1kg] before and after ischemia plus A1 receptor antagonist [2.25 / 1 kg] after ischemia. Ischemia induced by clamping of common carotid artery and the drugs was injected subsequently into peritoneum after reduction of inflammation of ischemic zone. The Y-maze memory test performed after completing the treatment period, afterward brains fixed and prepared for microscopic nissl staining method. The counting of pyramidal cells were performed at 53500 square micrometer of CA1. Data were analyzed using SPSS-15 and ANOVA test. The Y-maze test showed extensive de?cit in short-term memory in ischemic group [PA=200] but in treatment groups this deficit significantly reduced [PA=243, 248 and 265]. The normal neuronal cell in ischemic group was significantly lowered [n=87] than treatment groups [n=111, 105 and 125] including ascorbic acid group [125], adenosine receptor agonist [105] and ascorbic acid plus agonist adenosine receptor [111]. The number of normal neuronal cell in ischemic groups significantly is reduced compared to treatment group [P<0.05]. This study showed that concurrent treatment of ascorbic acid and Adenosine A1 receptor agonist can significantly reduce the complications caused by brain ischemia in CA1 area of hippocampus

[Effect of Neurotrophin-3 on differentiation of rat hair follicle stem cells into neural like cells]

The aim of this study was to evaluate the effect of NT-3 on the decrease of the differentiation time of bulge stem cells of rat hair follicle from neuron like cells. The bulge region of the rat whisker was isolated from and cultured in DMEM/F12 supplemented with epidermal growth factor [EGF] in 3 groups for 7,8 and 9 days. Then 10 ng /ml NT-3 was added to each group for 3 days. Finally, cell differentiation was assessed by immunocytochemistry using betaIII-tubulin antibody and the result was compared with control groups. According to our results hair follicle bulge stem cells expressed CD34 and Nestin in 7-9 and 10-12 first days after cultivation respectively. By using NT-3 duration differentiation process the cells showed positive reaction to betaIII-tubulin antibody. The results indicate that NT-3 can affect on differentiation speed even in less than 10 days

effect of garlic extract on expression of INF gamma and inos genes in macrophages infected with leishmania major

The study was aimed to show the effect of molecular mechanism of Aqueous Garlic Extract [AGE] on expression of IFN gamma and iNOS genes in Leishmania major. Leishmania major promastigotes [MRHO/IR/75/ER] were added to the in-vitro cultured J774 cell line, the cells were incubated fro 72 hours. Various concentrations of garlic extract [9.25, 18.5, 37, 74, 148 mg/ml] were added to the infected cells. MTT assay was applied for cellular proliferation. After 72 hours of incubation, supernatants were collected and total RNA was extracted from the infected cells. The express of IFN gamma and iNOS gene were studied by RT-PCR method. The colorimetric MTT assay after 3 days of incubation showed cytotoxic effect of garlic extract with an IC50 of 37 mg/ml. In addition, IFN gamma and iNOS genes expression by RT-PCR indicated that garlic extract lead to over expression of these genes in 1774 cell line infected with L. major. Garlic extract exerts cytotoxic effect on infected J774 cell line. In addition, the hypthesis that garlic can improve cellular immunity with raising the expression of IFN gamma and of iNOS genes confirmed

[ role of aqueous garlic extract [Allium sativum] on interleukin 10 and 12 in leishmania infected macrophages]

Leishmaniasis is a disease produced by several species of protozoa of the Leishmania genus. As a therapeutic agent, garlic [Allium sativum] is one of the most popular plants in traditional medicinal systems and is increasingly becoming important in herbal alternative therapy. The aim of the present study was to investigate the effects of aqueous garlic extract [AGE] on enhancing T-helper IL-10 and IL-12 cytokines in the culture via macrophages for engulfment and destruction of Leishmania. After proliferation of macrophages in culture and incubation with Leishmania during 72 hours, we added aqueaous garlic extract by several doses 9.25, 18.5, 37, 74 and 148 micro g/ml for 18, 24 and 48 hours. Trypan blue and MTT tests were carried out in order to test the safety of extracts and j744 cells treated with them, as well as cell viability. An enzyme-linked immunosorbant assay [ELISA] was performed on macrophages for interleukin IL-10 and IL-12. The results suggested that garlic extract may provide effective therapy against Leishmania. Our results showed that dose of 37 micro g/ml for 48 hours of garlic extract destructed promastigots by IL-12 secreted from macrophages. Based on the results of this study, IL-12 is crucial for defense against parasitic pathogens. IL-12 administration differentially affected immune response to invading Leishmania parasites

[Neurotrophic effects of silibinin on differentiation of hair follicle stem cells to neurons]

Hair Follicle Bulge region due to its availability and abundance is one of the areas which is easily accessible to Multi-potent stem cells that expresses Nestin marker [neuronal stem cells protein]. Stem cells bulge region in hair follicle stem cells has high potency to be differentiated to neuronal cells. Silibinin as an active component of Silybum marianum has anti-oxidant, anti-inflammatory, anti-carcinogenic, hepatoprotective, neurotrophic and neuroprotective effects. The aim of the present study was to evaluate the neurotrophic effects of silibinin on differentiation of hair follicle stem cells to neurons. Bulge area of whiskers in Rat was isolated and cultivated three weeks in supplemented DMEM/F12 and epidermal growth factor [EGF]. Then the cells were exposed over the concentrations of 0.05microg/ml, 0.1microg/ml, 0.4microg/ml, 0.5microg/ml, 0.7microg/ml Silibinin and Neurotrophin-3. Two weeks after culture, plated bulge cells were immunostained with Nestin and differentiated stem cells were immunostained with beta III tubulin by immunocytochemistry techniques. The results were evaluated by T-test student analysis. A Pvalue less than 0.05 was considered significant. The nestin marker was clearly demonstrated in bulge regions during the first week, but after two weeks, parallel to stem cells differentiating neuronal cells, beta III tubulin marker was expressed in neuronal cells. The toxic effects of 1microg/ml Silibinin on stem cells were also demonstrated, and it stopped the cell growth at the end of the first week. The maximum differentiation on stem cells in 0.5microg/ml Silibinin was observed to be significant [P<0.05]. Silibinin concentration increase led to reduced differentiation. Silibinin with neurotrophin 3 increased the differentiation of stem cells. Silibinin concentrations of 1microg/ml and more have toxic effects on hair follicles stem cell differentiation. Also, silibinin concentrations less than 0.1microg/ml had no effect on proliferation and differentiation hair follicle stem cells. Whereas 0.5microg/ml concentrations had significant effects on the differentiation processes of hair follicle stem cells to neuron

[Long-term complications of enucleated or severely traumatized eyes in war veterans]

To evaluate anophthalmic socket complications and the incidence of sympathetic ophthalmia among individuals who had undergone primary enucleation or severe ocular trauma during the war between Iran and Iraq. All monocular veterans of the mentioned war in Khorasan province, Iran were recalled during a 7-month period. Preliminary data including age at the time of injury, occupation, previous operations and ocular symptoms were evaluated and all participants underwent a complete ophthalmologic examination particularly regarding the anophthalmic socket, orbital implant and prosthesis condition. Overall, 135 male individuals participated in this study. Mean age was 42 +/- 7 years and 86% were 30-50 years. Patients had undergone complete enucleation in 39 [28.9%], partial enucleation in 21 [15.6%] and evisceration in 6 [4.4%] cases. In 34 cases [25.2%] pthisis bulbi had occurred after trauma and the type of surgery was not identifiable in the remaining 35 cases [25.9%]. The most common symptom in injured eyes was mucoid or mucopurulent discharge [71%]. Common complications in 101 subjects with previous operations were superior sulcus deformity [72.3%] and socket contracture [44.5%]. Socket motility was satisfactory only in 18%. All signs of the anophthalmic syndrome are more severe and more prevalent among enucleated cases secondary to war injuries. Due to the rarity of sympathetic ophthalmia, we suggest enucleation and orbital implantation in an elective setting

[Study of differentiation of hippocampal stem cells into rod photoreceptors in rat]

Vision is one of the most important organs in the body. Damage to this organ causes a severe disability in humans. In retinitis pigmentosa, the degeneration of photoreceptors causes blindness. So far, more than 100 mutations have been detected in photoreceptors, which they result in opsin malfunction. The aim of present study is to differentiate the hippocampal stem cells of rat to the rod photoreceptors. Stem cells of the hippocampus were obtained from rat embryos, 18 days of age [E18]. Pregnant female rats were killed and the head of their embryos were separated. Then, the embryos' hippocampus was removed according to Banker's method. Hippocampal cells were dissociated by Fish Bach's method. The cells were cultured in flasks [25cm[2]]. After 3 days, the cells were isolated by trypsin, counted using trypan blue and hemocytometer. Cell suspensions were prepared in two cell concentrations; high [2x10[5]cells/ micro l] and low [2x10[4] cells/ micro l] concentration, then, cultured using DMEM/F12 supplemented with fetal bovine serum 10% [FBS] in six wells plates. Prior to culture of the cells, the first and second row of plates were coated with poly L-lysine and inactivated astrocytes, respectively. Following incubation of the plates at 37°C for 4 days, different concentrations of All Transe Retinoic Acid [ATRA] and 9-CIS RA were added daily for 6 days, and finally immunocytochemistery was carried out using anti-opsin monoclonal antibody. The results of current study showed that the plates, which are respectively treated with ATRA and 9-CIS RA in a concentration of 500nM and 100nM for 6 days had the most differentiated cells. In addition, maximum differenced cells were observed with 100nM of 9-Cis RA. The differentiated cells were detected in wells, which were only coated with inactivated astrocytes in either a high or low concentration of cell suspension. These findings indicated that inactivated astrocytes as a feeder layer and extrinsic factors such as [ATRA] and 9-Cis RA increase differentiation of hippocampal stem cells into rod photoreceptors

Effects of retinoic acid and astrocyte as a feeder layer in differentiation of hippocampal stem cell

Adult stem cells are pluripotent cells conventionally isolated from some part of body by different methods. From developmental stand point, murine neural stem cells represent an accessible and important system for studies of basic stem cell property such as self- renewal and multipotency. In this study hippocampal stem cells obtained from embryonic day 18 [E18]. Pregnant female rats were killed, embryos heads separated and then hippocampus isolated by the method of Banker, then their cells dissociated by the methods of Fishbach, and plated in flask 25cm, after 3 days cells separated by tripsin, counted with trepan blue and hemocytometer, divided into two density [high 200000] and [low 20000 cells]. Before transplantation of cells, six well plates coated with poly L lysin and inactivated astrocyte. Then isolated cells transplanted into 6 well plate for 4 days with medium DMEM/F12 supplemented with FBS10%. After 4 days different doses of ARTA and RA cis-9 added per well for 6 days, and then immunocytochemistery were done. After 6 days of treatment with above factors, doses of 100nM RA cis-9 and 500 nM ATRA have the more staining cells with monoclonal antibody. But in 100 nM RA cis-9, we saw maximum differentiated cells. All of differentiation were done on wells with inactivated astrocyte layer in high and low density. Inactivated astrocyte as a feeder layer and extrinsic factors such as All Transe Retinoic Acid [ATRA] and RA cis-9 can cause differentiation in hippocampal stem cells into photoreceptor like cell

[Effects of retinoic acid and astrocyte as a feeder layer in differentiation of hippocampal stem cell]

Background: Adult stem cells are pluripotent cells conventionally isolated from some part of body by different methods. From developmental stand point, murine neural stem cells represent an accessible and important system for studies of basic stem cell property such as self- renewal and multipotency

Materials and Methods: In this study hipocampal stem cells obtained from embryonic day 18 [E18] Pregnant female rats were killed, embryos heads separate and then hippocampus isolated by the method of Banker, then their cells dissociated by the methods of Fishbach, and plated in flask 25cm, after 3 days cells separated by tripsin, counted with trepan blue and hemocytometer, divided into two density [high 200000] and [low 20000 cells]. Before transplantation of cells, six well plates coated with poly L lysin and inactivated astrocyte, Then isolated cells transplanted into 6 well plate for 4 days with medium DMEM/F12 supplemented with FBS10%. After 4 days different doses of ARTA and RA cis-9 added per well for 6 days, and then immunocytochemistery were done

Results: After 6 days of treatment with above factors, doses of 100nM RA cis-9 and 500 nM ATRA have the more staining cells with monoclonal antibody . But in 100 nM RA cis-9, we saw maximum differentiated cells. All of differentiation were done on wells with inactivated astrocyte layer in high and low density

Conclusions: Inactivated astrocyte as a feeder layer and extrinsic factors such as All Transe Retinoic Acid [ATRA] and RA cis-9 can cause differentiation in hippocampal stem cells into photoreceptor like cell

Stages of development of renal glomeruli in the newborn rat kidney

Glomerular development of the kidney was studied in newborn rats by electron microscopy. Four different stages of glomerular development were defined: vesicle formation, S-shaped body stage, capillary loop formation, and glomerular maturation. In the first stage, the mesenchymal cells form a spheroid mass. This is followed by the S-shaped body stage in which clefts appear in the mass. Afterwards, capillary loop formation, junctional migration, podocyte differentiation, and interdigitation of epithelial processes occur. Finally, the cytoplasm of endothelial cells becomes thinner. The urinary space is visible in this stage. The fusion of epithelial and endothelial basement membranes results in formation of the layers of the GBM. The increase in the number of podocyte processes and endothelial cell fenestrations are important events in the maturation phase

role of T-lymphocyte subpopulation in renal allograft rejection

Twenty-two recipients of HLA-nonidentical living related and non- related renal allografts were studied for alterations in the relative percentage of OKT4-positive peripheral blood T-cells after transplantation. Characteristic shifts in the ratio of T-helper to T-suppressor/cytotoxic cells [TH/TS-C], but not absolute cell numbers, were demonstrated to correspond with the status of the allograft. Our results are indicative of a correlation between rejection episodes and the increase in OKT4:OKT8 ratios, that were characterized by a significant rise in the percentage of OKT4-positive cells [P=Zero.1], and a decrease in the percentage of OKT8-positive cells [P=Zero.1]