Neuronal cell reconstruction with umbilical cord blood cellsi n the Bbain hypoxia-ischemia

Brain hypoxia-ischemia is a human neonatal injury that is considered a candidate for stem cell therapy. The possible therapeutic potential of human umbilical cord blood [HUCB] stem cells was evaluated in 14-day-old rats subjected to the right common carotid occlusion, a model of neonatal brain hypoxiaischemia. Seven days after hypoxia-ischemia, rats received either saline solution or 4 × 105 HUCB cells i.v. Rats in control group did not receive any injection. After two weeks, rats were assessed using two motor tests. Subsequently, rats were scarified for histological and immunohistochemical analyses. Our immunohistochemical findings demonstrated selective migration of the injected HUCB cells to the ischemic area as well as reduction in infarct volume. Seven days after surgery, we found significant recovery in the behavioral performance in the test group [12.7 +/- 0.3] compared to the sham group [10.0 +/- 0.05], a trend which continued to day 14 [15.3 +/- 0.3 vs. 11.9 +/- 0.5, P<0.05]. Postural and motor asymmetries at days 7 and 14 in the test group showed a significant decrease in the percentage of right turns in comparison to the sham group [75% and 59% vs. 97% and 96%, P<0.05]. The results show the potential of HUCB stem cells in reduction of neurologic deficits associated with neonatal hypoxia-ischemia.

Single dose effect of diazinon on biochemical parameters in testis tissue of adult rats and the protective effect of vitamin E

Diazinon [DZN] is an organophosphate pesticide that widely used for agricultural pest control all over the world. DZN affects target organs including reproductive system by inhibiting the activity of acetylcholinesterase and inducing oxidative stress. Vitamin E [alpha-tocopherol] is a strong antioxidant which inhibits free radicals, and probably can reduce lipid perxidation effectively in biological systems. The present study, aimed to evaluate the effects of DZN on malondialdehyde [MDA] and glutathione [GSH] levels in testis of rats and protective effect of vitamin E. In this experimental study, thirty adult male Wistar rats [200-250 gr] were divided into 5 groups [n= 6]: control group [did not receive any material], sham group [received only pure olive oil], experimental group 1 [DZN, 60 mg/kg], experimental group 2 [Vit E, 200 mg/kg] and experimental group 3 [DZN+Vit E, with the same dose]. All groups were sacrificed after 6 weeks and right testis was used to measure the MDA and GSH levels. The amount of MDA was determined by the thiobarbituric acid assay and 5, 5-Dithio-bis [2nitrobenzoic acid] DTNB-recycling protocol was used for GSH assay. The results showed that DZN increased MDA level [p<0.001] and reduced GSH level [p<0.001]. Administration of DZN plus vitamin E decreased the MDA level [p<0.001] and increased GSH level [p=0.001]. DZN induced lipid peroxidation in the testis of rats. Vitamin E by its antioxidant activity was able to improve the toxic effect of DZN

[Assessment of erythrocyte and serum cholinesterase activity in male rats treated with diazinon, and protective role of vitamin E]

Background: Diazinon [DZN] is an organophosphate [OP] that inhibit cholinesterase activity and induce oxidative stress in the target tissues such as the the reproductive system .The aim of present study was to evaluate the effects of diazinon on cholinesterase activity in blood serum and erythrocytes of male rats and to assess the protective role of vitamin E.

Material and Methods: In this experimental study, thirty adult male wistar rats [200 - 250 gr] were divided into 5 groups [n = 6]: control group [did not receive any material], sham group [received only pure olive oil], experimental group 1 [DZN, 60 mg/kg], experimental group 2 [received DZN+Vit E daily, with the same dose] and experimental group 3[Vit E 200 mg/kg]. DZN and solvent was injected intraperitoneally [IP] and vitamin E was administrated by gavage. After 6 weeks, 3 ml blood from the heart tissue was taken and titrimetric and Ellman's method respectively was used for serum and erythrocyte cholinesterases activity evaluation. Data were analyzed using SPSS software. P<0.05 value was taken as statistically significant.

Results: The results showed that diazinon, reduced activity of acetylcholinesterase [P= 0.000] and pseudocholinesterase [P=0.01] in experimental group 1 compared to the control group. Inhibition in erythrocyte and serum cholinesterase activity was not recovered in experimental group 2 [use of diazinon plus vitamin E diazinon].

Conclusion: According to inhibit the activity of two important enzymes of erythrocyte and serum cholinesterase in rats treated with diazinon, there are possibility of cytotoxicity at farmers and people who are in contact with these compounds. On the other hand, cholinesterase activity did not recovered by vitamin E probably due to the competitive nature of enzyme inhibition by diazinon. Therefore, care should be taken to prevent from entering toxin to the body.

effect of maternal nicotine on basement membrane collagen IV of brain microvessels changes in neonatal Balb/C mice

Nicotine can pass through placental blood barrier and accumulate in the developing organs of fetus. Also, entering the breast milk, nicotine can have an effect on the neonates. Investigations have showed that collagen IV is one of the most important micro vessels basement membrane components. In this study, the effect of maternal nicotine exposure in pre and postnatal periods on collagen IV in microvessels of neonatal Balb/C mice brain cortex was studied by immunohistochemistry technique. 24 pregnant Balb/C mice were divided in to 4 groups [6 mice in each group]: two experimental and 2 control groups. The mothers in the 1st experimental group were injected 3 mg/kg nicotine intrapritoneally from the 5th day of pregnancy to parturition daily and in 2nd experimental group the same procedure was repeated to the 10th day after parturition [lactation]. The control groups received the same volume of normal saline during the same time. 10 days after delivery, the brain tissues of newborns were isolated. Then, prepared blocks from fixed brain were cut serially for immunohistochemical assay. The findings of the present study indicated that collagen IV reaction in microvessels basement membrane in the first experimental group increased significantly compared to the first control group [p=0.002]. In addition, collagen IV reaction in microvessels basement membrane in the 2nd experimental group increased significantly compared to the 2nd control group [p=0.002]. However, no significant difference was observed between the two experimental groups. These results suggested that maternal nicotine exposure during prenatal period may increase basement membrane collagen IV expression. Also, nicotine increases in maternal breast milk has no effect on basement membrane collagen IV expression

Evaluation of laminin expression during mouse lens development

Among the components of the extracellular matrix [ECM] and basement membrane [BM], laminitis heterotrimeric glycoprotein [laminin] and collagen type IV are the most important. In a previous study we have examined the role of collagen type IV in the developing lens capsule. The present study aims to determine the appearance and distribution of laminin in the BM and ECM of lenses during visual system morphogenesis. Pregnant Balb/C mice were randomly selected and maintained under normal conditions. The presence of a vaginal plug was assumed as day zero of pregnancy. From embryonic days 11 to 20, pregnant animals were sacrificed and their fetuses were collected for histotechnical prossessing. Our data revealed that laminin appeared during the early stage of gestation [day 12] in the BM of the anterior epithelial lens cells. The amount of laminin gradually increased in the ECM and posterioir lens capsule epithelial cells until days 14-18. After this period, a strongly positive laminin reaction was not observed in any part of the lens structure. These findings establish the importance of the laminin molecule in the developing optic cup [OC] and lens differentiation. It could be assumed that any changes in the presence of laminin during the critical period of eye development may result in visual system defects such as cataracts or congenital eye abnormalities

Effects of Vitamin-E treatment on CatSper genes expression and sperm quality in the testis of the aging mouse

CatSper genes are a novel family of four sperm-specific calcium channels, which indicate testis-specific expression patterns. Despite the crucial role of CatSper genes in the male reproduction, very little is known about the factors that regulate their expression. The objective of this study was to investigate the effects of vitamin E treatment on the expression of CatSper 1 and CatSper 2 genes as well as sperm quality in the aged male mice. Twenty four 11-12 months old aged male mice and twenty four 2-3-months old young male mice were randomly divided into four groups. Control groups received no injection. The experimental groups of male mice were received intraperitoneal injection of 106 mg/kg vitamin E daily for 35 days. Left testis and cauda epididymides from each mouse were collected on the days 21, 28 and 35 following vitamin E treatment and were used for Real-Time PCR and immunohistochemistry. Also, sperm analysis was performed according to the WHO guidelines given for human sperm examination. Data were analyzed using SPSS software Administration of vitamin E improved sperm parameters in the aged as well as young adult male mice. In addition, the expression of CatSper genes increased following vitamin E treatment. Also, intensity of signal for CatSper1 and CatSper2 increased in the head and middle piece of sperm in experimental group as compared to those of control ones. The vitamin E treatment significantly improved the sperm quality, especially in terms of sperm motility, count and morphology rate. Furthermore, CatSper genes expression could be up-regulated by the vitamin E treatment

Therapeutic potential of umbilical cord blood stem cells on brain damage of a model of stroke

Human cord blood-derived stem cells are a rich source of stem cells as well as precursors. With regard to the researchers have focused on the therapeutic potential of stem cell in the neurological disease such as stroke, the aim of this study was the investigation of the therapeutic effects of human cord blood-derived stem cells in cerebral ischemia on rat. This study was carried out on young rats. Firstly, to create a laboratory model of ischemic stroke, carotid artery of animals was occluded for 30 minutes. Then, umbilical cord blood cells were isolated and labeled using bromodeoxyuridine and 210[5] cells were injected into the experimental group via the tail vein. Rats with hypoxic conditions were used as a sham group. A group of animals did not receive any injection or surgeries were used as a control. Obtained results were evaluated based on behavioral responses and immunohistochemistry, with emphasis on areas of putamen and caudate nucleus in the control, sham and experimental groups. Our results indicated that behavioral recovery was observed in the experimental group compared to the either the sham or the control group. However, histological studies demonstrated a low percent of tissue injury in the experimental group in comparison with the sham group. Stem cell transplantation is beneficial for the brain tissue reparation after hypoxic ischemic cell

immunohistochemical study of retinal collagen IV expression during pre- and postnatal periods in Balb/c mice

Basement membranes are specialized extracellular matrices which play important roles such as cell regulation, proliferation and migration. Collagen fibers, especially type IV, are the most important basement membrane constituents. As retina is one of the target organs in diabetes mellitus, and nephropathy is a major cause of end stage renal and retinal diseases resulting in increased morbidity and mortality, early diagnosis leads to better treatment. Hence, in this investigation, the appearance and distribution of collagen IV during gestational days and early postnatal periods were observed. 24 intact female Balb/c mice were kept under normal conditions. After mating, appearance of a vaginal plug was assumed as day zero of the pregnancy. From days 13-18 of gestation, the pregnant mice were euthanised and their embryos as well as pups from days 1 to 5 were collected. For histochemichal studies, heads of the specimens were fixed, serially sectioned and immunohistochemi-call studies were performed by using monoclonal antibodies for tracing of collagen type IV. Our findings revealed that the amount of collagen IV in the internal limiting membrane [ILM] and extra cellular matrix [ECM] of the retina, as well as vessels of the vitreus body appear on embryonic day 16. Also, a patchy distribution was observed in the pig-mented epithelium which continued to further develop until the end stage of embryonic life. Strong labeling was observed until postnatal day 3 but did not increase significantly thereafter. These findings establish the importance of collagen IV during the critical period of retinal development. In addition, this study indicates that high levels of collagen IV are present in the basal membrane [BM] of the inner limiting membranes and pigmented epithelium 3[rd] post natal on the 3[rd] postnatal day

Maternal nicotine induces collagen type IV changes in the mice lung parenchyma and its vessels

One of the undesirable effects of maternal nicotine exposure during pregnancy is pulmonary hypertension. Since nicotine binds to its receptors on pulmonary vessels the hypothesis of this research was the possible structural changes that nicotine may cause on newborn vessels. Twenty-four female BALB/c mice were mated and finding vaginal plug was assumed as day zero of pregnancy. Pregnant mice were divided into 2 experimental and 2 control groups. Experimental group 1 received 3 mg/kg nicotine intraperitoneally from day 5 of gestation until the last day of pregnancy. Experimental group 2 received the same amount of nicotine during the same gestational days as well as the first 2 weeks after birth [lactation]. The control groups received the same volume of normal saline during the same periods. At the end of exposure times, all the newborns [experimental and control] were anesthetized, their lungs were removed and immunohistochemical studies were carried out for tracing collagen. Our findings indicated that collagen reaction in the bronchial basement membrane [BBM] and extracellular matrix [ECM] of the lung parenchyma in experimental groups increased significantly compared to the control groups but these changes were not observed in BM of lung vessels in the experimental groups. These data indicate that nicotine exposure during pregnancy does not cause a significant change in collagen type IV in BM of lung vessels. But this does not mean that other types of collagen fibers do not indicate change because the wall thickness of pulmonary vessels in experimental groups increased significantly compared to the control groups

Effects of crude onion extract on murine testis

Ancestral beliefs have tied onion consumption and its medicinal properties to increases in sexual desire. The present study tries to put this old belief to test. Therefore, this study has explored the effects of oral consumption of onion extract on the reproductive activity and testicular structure of adult male mice. In this study, twenty-four 40-day old male Balb/C mice were randomly divided into two experimental and control groups. The experimental group received daily doses of onion [Allium cepa] extract [lm/100gr/B.W.] for 10 days. The controls received the same volume of normal saline. At the end of the study, all the animals were anesthetized and scarified by cervical dislocation and their testes were harvested for histological study. For statistical analysis, t-tests were used to compare the means of the two groups while the significance level was set at p<0.05. The mean internal diameter of seminiferous tubules showed a significant increase in the experimental [56.32 +/- 1.42micro m, p<0.005] relative to the control group [31.27 +/- 3.16], whereas their mean external diameters did not show any significant difference. In addition, proliferation of spermatogonial cells and spermatocytes I and II increased significantly in the experimental group [264 +/- 14.11, p<0.0005]. It seems that administration of onion extract affects both structure and proliferation of cells in tubules and enhances spermatogenesis in the murine

role of type IV collagen in developing lens in mouse fetuses

Extracellular matrix [ECM] and basement membrane [BM] play important roles in many developmental processes during development and after birth. Among the components of the BM, collagen fibers specially type IV are the most important parts. The aim of this study was to determine the time when collagen type IV appears in the BM of lens structure during mouse embryonic development. In this experimental study, 22 female Balb/C mice were randomly selected and were kept under normal condition, finding vaginal plug was assumed as day zero of pregnancy. From embryonic day 10 to 20, all specimens were sacrificed by cervical dislocation and their heads were fixed, serially sectioned and immunohistochemistry study for tracing collagen type IV in lens were carried out. Our data revealed that collagen type IV appeared at the early stage of gestation day 12 in BM of anterior epithelial lens cells and the amount of this protein gradually increased until days 15-17 in ECM and posterior capsule epithelium. After this period, severe reaction was not observed in any part of the lens. These findings establish the important role of collagen IV in developing optic cup and any changes during critical period of pregnancy may be result in severe visual system defect

Unique carbohydrate appearance of the floor plate during early neurolation

During early neurogenesis, the floor plate plays an essential role[s] in the differentiation of the ventral portion of neural tube. In this study, we detected the specific distribution of unique glycoconjugate during the floor plate differentiation. Formalin fixed paraffin sections of BALB/c mice [10-14 embryonic days] were processed for histochemical studies using horseradish peroxidase-labelled N-acetylgalactosamine [GalNAc] sensitive lectins. Histochemical analysis has revealed the presence of unique Wisteria floribunda [WFA]-sensitive glycoconjugate reaction in the floor plate area and surrounding extracellular matrix. Weak reaction also was observed in the outer surface of the basal zone of the neural tube. Extensive differences among the GalNAc lectins were demonstrated during the sensitive time of motoneuron differentiation. There was no reaction with other tested GalNAc lectins. The duration and distribution of WFA lectin reactions suggest that these molecules may play a key role[s] in tissue interactions and subsequent formation of the adjacent tissues including floor plate and basal plate differentiation during critical morphogenic period. Furthermore, our findings indicate that among considered early neuronal morphogenic [embryonic] days, WFA reactions were increased and expanded near the end of gestation period