The protective effect of methanolic extract of Verbascum cheiranthifolium and Biebersteinia multifida DC on hippocampus damage induced by diazinon in male Wistar rats: An experimental study.

Diazinon (DZN) an organophosphate (OP), with the most important mechanism of action of DZN being induction of oxidative stress (OS) and inhibition of the enzyme acetylcholinesterase (AChE). Verbascum cheiranthifolium (VER) and Biebersteinia multifida (BM) belong to the Scrophulariaceae and Biebersteiniaceae family respectively. These plants are widely used in Iranian traditional medicine due to their beneficial effects. Thus, this research aimed to appraise the protective effects of the methanolic extract of the VER and BM on changes in the level of expression of α7 and α4 subunits of nicotinic acetylcholine receptors (nAChRs) in hippocampus (HPC) of DZN-treated rats. In this research, 36 male Wistar rats were used and randomly divided into six groups: Control, DZN (40 mg/kg), VER (1 g/kg), DZN+VER (40 mg/kg+1 g/kg), BM (150 mg/kg), and DZN+BM (40 mg/kg+150 mg/kg). At the end of treatment periods, the animals of all groups underwent the Morris water maze (MWM) test. The rats were anesthetized, and blood sampling was performed. Eventually, the brain was removed for histological study and evaluation of OS parameters. The results indicated that DZN increased the extent of expression of nAChRs in the HPC and significantly inhibited cholinesterase (ChEs) activity plus OS parameters. Also, in MWM, the time to find the platform was significantly longer in the DZN group, while the time and the distance in the probe test were lower than in the control groups. VER and BM extract in the treatment groups simultaneously improved the extent of expression of nAChRs, ChEs activity, as well as the parameters of OS and spatial memory significantly. In conclusion, our results support the neuroprotective properties of VER and BM extract versus DZN in rats. Accordingly, the extracts of VER and BM may be useful as an approach for the treatment of learning disorders and memory enhancement.

Inhibition of Autophagy in Heat-Stressed Sperm of Adult Mice: A Possible Role of Catsper1, 2 Channel Proteins.

Objective: Various phenomena guarantee gamete maturation and formation at all stages of evolution, one of which is autophagy playing a critical role in the final morphology of gametes, particularly sperms. Autophagy is influenced by oxidative stress, disturbances of calcium homeostasis, and hyperthermia conditions. The current study aimed to assess the autophagy-related proteins along with the activity of sperm calcium channel (CatSper) proteins following the induction of heat stress (HS). Methods: The study sample includes two groups of adult mice: sham and HS groups. In the HS group, the right testis was transferred to the abdominal cavity for 120 hours and then returned to the scrotum where it remained for 7 days. After 7 days, the testis and epididymis were removed to conduct real-time, immunohistochemical studies, sperm parameter evaluation, and seminiferous tubule assessment. In this study, the expression and distribution of autophagy proteins were measured. Plus, CatSper1 and CatSper2 were evaluated as proteins of calcium channels. Results: The results of the present study demonstrated that the expression intensity of autophagy indices in seminiferous tubules decreased significantly after HS induction, which was associated with a decrease in the distribution of CatSper proteins in the sperms. HS led to morphological changes in sperm, reduced motility and viability of sperm, and decreased spermatogenesis indices. Conclusion: In this study, following heat stress, the decrease in CatSper protein distribution may lead to the structural disorder of CatSper channels, which could strongly affect autophagic activity. Also, disruption of spermatogenesis and sperm parameters may be the consequence of decreased autophagy activity.

Evaluation of sperm chromatin/DNA integrity, morphology, and Catsper expression on diabetic C57BL/6 mice.

BACKGROUND: Diabetes is associated with reproductive impairment on the male reproductive system and causes complications such as decreased libido, fertility, spermatogenesis, sperm motility, and morphology. High levels of blood sugar may affect sperm quality and reduce the potential for male fertility. Increased levels of sperm DNA damage is often associated with reduced count and motility or abnormal morphology. METHODS: This experimental study was conducted in Mashhad University of Medical Sciences. In this work, 40 mice (C57BL/6) were divided randomly into 4 groups: 1) Control, 2) Diabetic, 3) Diabetic + Insulin, and 4) Sham. After 35 days, the right epididymis of all specimens was used for Real-Time PCR and left epididymis for evaluation of sperm parameters using Aniline blue, Toluidine blue, Papanicolaou, and immunohistochemical study. Also, testes were applied for immunohistochemical, TUNEL studies, and biochemical assay. RESULTS: Results of this study showed that chromatin integrity, morphology, cation channels of sperm (Catsper) expression, and biochemical factors level were significantly changed in diabetic mice in comparison to other groups (P<0.05) and treatment with insulin improved these parameters. CONCLUSION: Our findings showed that the sperm parameters such as DNA integrity, morphology, and Catsper expression change in diabetic mice.

Autophagy comparative after decompression of tunica albuginea in testicular torsion in mature and immature rat.

BACKGROUND: One of the types of catabolic processes in cells is autophagy, which maintains cell homeostasis. It is also known as a defense response to oxidative stress. This study aimed to track the autophagic activity of testicular tissue in mature and immature rats in 3 conditions: (1) physiologic, (2) under increased pressure in the tunica albuginea compartment following the induction of testicular torsion, and (3) decompression of this compartment by flap technique. METHODS: This study was performed on 72 Wistar rats divided into 3 mature and 3 immature groups, each of the groups including Sham group, testicular torsion induction group for 6 hours (Torsion Detorsion), and testicular torsion induction group for 6 hours, followed by the flap technique. RESULTS: In the physiologic state or Sham, autophagic indices (LC3, Beclin-1) are active, and more activity was observed in mature rats, compared with immature ones, which indicated the activity of this process during development and maturity. In the Torsion Detorsion group, significant expression of autophagy and apoptosis indices (caspase3) was observed along with the degradation of spermatogenesis and steroidogenesis. It seems that in this state, the activation of autophagy leads to programmed death and is in line with apoptosis. Finally, in the flap technique group, an increase in the expression of autophagy indices was accompanied by a reduction in the expression of apoptotic indices and improved spermatogenesis and steroidogenesis. CONCLUSION: Decompression of tunica albuginea by flap technique can activate of autophagy process to save tissue function and eliminate stress caused by testicular torsion.

Evaluation of Sertoli cell autophagy associated with laminin, fibronectin, and caspase-3 proteins' alteration, following testicular torsion rat.

Autophagy is a vital process that maintains cellular homeostasis by joining lysosomes and providing energy production substrates. In testicular tissue, Sertoli cells play functional roles in spermatogenesis and steroidogenesis. It is well known that autophagy physiologically occurs in the Sertoli cells. Under pathological conditions, such as testicular torsion, autophagy can be activated under high-stress stimuli. It is worth noting that Sertoli cells receive autophagy-induced signals through some extracellular matrix proteins, e.g. laminin and fibronectin. The present study aims to evaluate Sertoli cells' autophagy-associated extracellular matrix proteins' alteration following testicular torsion in rat model. The animals were divided into two groups as sham and testicular torsion/detorsion groups. In the testicular torsion/detorsion group, testicular torsion was maintained for 6 hr, followed by detorsion for 14 days. The obtained results revealed that testicular torsion-induced oxidative stress leads to increased autophagy in Sertoli cells as well as the whole testicular tissue. Moreover, extracellular matrix proteins including laminin and fibronectin act as autophagy-regulating proteins, in which their expression levels are reduced and increased respectively. In addition, the level of caspase-3, as an autophagy inhibitory protein, did not increase significantly in the cytoplasm of Sertoli cells as opposed to whole testicular tissue, indicating that autophagy is active after testicular torsion in these cells.

Diabetes up-regulated collagen IV and laminin α5 genes in mRNA and protein levels in seminiferous tubules of C57BL/6 adult mice.

Diabetes is a disease associated with impairment of the male reproductive system that causes complications such as decreased testosterone, the diameter of the seminiferous tubule, libido, and fertility. Extracellular matrix (ECM) molecules are involved in testicular development and spermatogenesis. Laminin and collagen are key proteins in seminiferous tubule basement membrane and play an important role in spermatogenesis. The present study was conducted to investigate the effect of diabetes on collagen IV and laminin α5 changes in mice testis. In this experimental study, 40 mice (C57BL/6) were divided randomly into 4 groups: 1) Control group: without intervention, 2) Diabetic group: treated mice with 50 mg/kg streptozotocin (STZ), 3) Diabetic + Insulin group: treated mice with STZ and insulin, and 4) Sham group: received citrate buffer. After 35 days, the left testes of all specimens were used for Real-Time PCR while their right testes were applied for immunohistochemical study and Periodic acid-Schiff (PAS) staining. This study showed that gene expression and immunoreactivity of laminin α5 and collagen IV were significantly increased in diabetic mice compared to other groups (P<0.05). Also, PAS staining showed the thickness of seminiferous tubule basement membrane in the Diabetic group compared to other group increased significantly (p<0.05). In Diabetic + Insulin compared to Diabetic group, gene expression, the intensity of immunoreactivity and thickness of seminiferous tubule basement membrane decreased significantly (P<0.05). Our findings indicated that diabetes causes up-regulation of collagen IV and laminin α5 in mRNA and protein levels in the seminiferous tubule basement membrane and may cause disorder in spermatogenesis in mice.

The protective role of alpha-lipoic acid on the appearance of fibronectin and laminin in renal tubules following diazinon exposure: An experimental immunohistochemical study.

Extracellular matrix (ECM) exerts a major role in maintaining the structure and developmental processes of tissues. To form the tubular basement membrane in the kidney, sulfate proteoglycans, collagen, laminin, fibronectin, and other glycoproteins congregate in the ECM. As an insecticide, diazinon (DZN) may alter the proportion of ECM by cholinesterase activity inhibition and oxidative stress. The naturally, alpha-lipoic acid (ALA) plays an effective and therapeutic role in the treatment of toxicities and diseases in the body. In the current study, an attempt was made to evaluate the impacts of alpha-lipoic acid on the distribution of fibronectin and laminin in the renal tubules of male Wistar rats following exposure to diazinon. In this study, the animal groups comprised 30 adult male Wistar rats (almost three months old) randomly distributed into the following groups; control, DZN (40 mg/kg), DZN + ALA (40 mg/kg+100 mg/kg), ALA (100 mg/kg), and sham. The rats were anesthetized after six weeks. Blood sampling was performed, and kidneys were removed for immunohistochemistry study. Diazinon reduced the distribution of fibronectin and laminin and significantly inhibited cholinesterase activity in the renal tubules. Furthermore, urea and creatinine levels were higher in diazinon than in other groups. ALA in the co-treatment group enhanced cholinesterase activity and distribution of both glycoproteins in the renal tubules. Urea and creatinine levels were meaningfully diminished in the DZN + ALA group. The nephrotoxic effect of diazinon in vivo was the reduced distribution of laminin and fibronectin, probably induced by cholinesterase activity inhibition. As an antioxidant with specific properties, ALA reduces the nephrotoxic effects of diazinon by multifarious mechanisms.

Evaluation of the anti-oxidant effect of ascorbic acid on apoptosis and proliferation of germinal epithelium cells of rat testis following malathion-induced toxicity.

OBJECTIVES: The aim of this study was to determine the protective role of ascorbic acid on apoptosis and proliferation of spermatogonia and primary spermatocyte cells after malathion administration as an organophosphate pesticide in rat testis. MATERIALS AND METHODS: Thirty male Wistar rats were randomly divided into five groups of 6 rats each, including control (no intervention), sham (normal saline 0.09%), malathion (50 mg/kg), malathion plus ascorbic acid (50 mg/kg and 200 mg/kg, respectively), and ascorbic acid (200 mg/kg) groups. Malathion and ascorbic acid were administrated via intraperitoneal injection once per day and seven times per week. After 6 weeks, animals were sacrificed, and testis tissue was used for evaluation of apoptosis and proliferation of germinal epithelium cells using the TUNEL and PCNA staining techniques. RESULTS: The results of TUNEL staining showed that the numbers of apoptotic cells in spermatogonia and primary spermatocyte cells were significantly increased in the malathion 50 mg/kg group vs control group (P<0.001). Co-administration of malathion 50 mg/kg and ascorbic acid 200 mg/kg significantly decreased the apoptotic cells in both cell types in comparison with malathion 50 mg/kg group (P<0.001). The results of PCNA staining revealed that the proliferation of these cells was significantly decreased in malathion 50 mg/kg group vs control group (P<0.001), and malathion 50 mg/kg plus ascorbic acid 200 mg/kg administration increased the proliferation of cells compared with malathion 50 mg/kg group (P<0.001). CONCLUSION: The results provide evidence that ascorbic acid showed preventive effects on malathion-induced toxicity in male rat testis.

The protective effect of alpha-lipoic acid on the expression of collagen IV, renal function, and oxidative stress induced by diazinon in the renal parenchyma of rat.

Diazinon (DZN) is an organophosphate pesticide that is commonly used in agriculture worldwide, including in Iran, and unfortunately, it leads to a variety of negative effects on the environment, animals, and humans. Alpha-lipoic acid (ALA) is an antioxidant agent that acts via scavenging of oxygen-free radicals. Collagen IV is a component of the main base membrane structure and DZN may also affect the expression of this key protein. The aim of this study was to evaluate antioxidant properties of ALA on the expression of collagen IV, renal function, and oxidative stress induced by DZN in renal tissue. In this experimental study, 30 adult male Wistar rats were randomly divided into five groups (n = 6) including: the control group, DZN (40 mg/kg) group, ALA (100 mg/kg) group, ALA (100 mg/kg) + DZN (40 mg/kg) group, and sham group. On day 0 and after 6 weeks, the urine and blood samples were collected to measure glomerular filtration rate (GFR). After 6 weeks, the rats were anesthetized and the left kidney was used for immunohistochemistry study and the right kidney was used to evaluate the oxidative stress parameters. The results have shown that ALA significantly improved the biochemical parameters including superoxide dismutase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, and GFR. In addition, ALA significantly prevented the expression of collagen IV that was changed by DZN administration in rats. We concluded that when exposed to DZN, depletion of antioxidant enzymes is accompanied by the induction of oxidative stress that might be beneficial in monitoring DZN toxicity and alpha-lipoic acid, as an antioxidant can overcome the toxicity induced by DZN in the kidney.

P53 expression in various types of hydropic placentas (through ploidy analysis as a complementary tool in diagnosis of samples).

BACKGROUND: Placentas characterized by hydropic swelling of chorionic villi occur in a spectrum of pathological conditions including hydropic abortion (HA), partial hydatidiform mole (PHM) and complete hydatidiform mole (CHM). The purpose of this study was to investigate whether the expression of p53 tumour suppressor protein could differentiate these various types of hydropic placentas. METHODS: p53 immunohistochemical staining was performed in 19 molar (8 PHM and 11 CHM) and 10 non-molar (HA) formalin-fixed, paraffin-embedded tissue samples. Ploidy analysis using flow cytometry was performed as a complementary tool in diagnosis of samples. RESULTS: DNA histograms obtained from all samples had confirmed diploidy in HAs and CHMs and triploidy in PHMs. p53 immunoreactivity was assessed in villous cytotrophoblasts, syncytiotrophoblasts and stromal cells. The p53 positive reaction was predominantly observed in the nuclei of cytotrophoblastic cells and rarely in stromal cells, no reaction was seen in syncytiotrophoblasts. The mean percentage of p53 positive cells were 6.10±3.75 for HA, 25.87±13.4 for PHM and 39.83±18.76 for CHM. There was a significant difference in P53 immunoreactivity of cytotrophoblastic cells between CHM and HA (P<0.001), and between PHM and HA (P=0.004). There was no significant difference in immunohistochemical reactivity between CHM and PHM (P=0.068). CONCLUSION: This study confirms that p53 immunostaining may be helpful in distinguishing complete and partial hydatidiform mole from hydropic abortion, but not complete hydatidiform mole from partial hydatidiform mole.

Evaluating the Protective Role of Ascorbic Acid in Malathion-induced Testis Tissue Toxicity of Male Rats.

BACKGROUND: Malathion is one of organophosphate pesticides that is widely used in agriculture and crops to control insects. Malathion affects body organs such as the reproductive system by inhibiting acetylcholinesterase activity and induction of oxidative stress. This study is aimed to investigate the effects of malathion on glutathione (GSH) and malondialdehyde (MDA) level in testis of male rats, as well as to study the protective role of Ascorbic Acid. METHODS: In this study, 30 adult male Wistar rats weighing approximately 200-250 g were divided into 5 groups of 6 rats each. These groups include a control group (no intervention), sham (normal saline 0.9%), experimental Group 1 (malathion 50 mg/kg), experimental Group 2 (Malathion 50 mg/kg + Ascorbic Acid 200 mg/kg), and experimental Group 3 (Ascorbic Acid 200 mg/kg). Malathion, solvents, and ascorbic acid were injected intraperitoneally. After 6 weeks, all groups were anesthetized, and the right testis was used to measure levels of MDA and GSH. MDA as a marker of lipid peroxidation and GSH content was used. RESULTS: The results showed that malathion increased MDA level and decreased GSH level compared with the control group (P < 0.001). It was also found that administration of malathion in combination with ascorbic acid reduced MDA level and increased the GSH level. CONCLUSIONS: Malathion-induced lipid peroxidation and oxidative stress in the testis of rats. In addition, it seems that ascorbic acid, due to its antioxidant capabilities, can improve malathion-induced poisonous changes.

The Effect of Diazinon on Cell Proliferation and Apoptosis in Testicular Tissue of Rats and The Protective Effect of Vitamin E.

BACKGROUND: Diazinon (DZN) is an organophosphate pesticide, and nowadays this pesticide is mostly used in agriculture. In this study, we analyzed the effects of DZN and vitamin E (Vit E) on apoptosis and the proliferation of germ cells in rat testis. MATERIALS AND METHODS: In this experimental study, 30 male Wistar rats were divided into five groups (n=6 per group) consisting of control, sham (received olive oil), experimental group i (60 mg/kg DZN), experimental group ii (60 mg/kg DZN and 200 mg/kg Vit E), and experimental group iii (200 mg/kg Vit E). After six weeks, left testis of rats was removed for the detection of proliferative cell nuclear antigen (PCNA) and terminal deoxynucleotidyl transferase end-labeling (TUNEL). RESULTS: Compared with the control group, DZN in the experimental group i decreased the number of PCNA-positive cells and increased the number of TUNEL-positive cells (P<0.001). Vit E improved detrimental changes by the decrease in the rate of apoptosis and the increase in the proliferation of testicular germ cells (P<0.001). CONCLUSION: Vit E can decrease the number of TUNEL-positive cells and increase the number of PCNA-positive cells by the neutralization of the toxicity caused by DZN in the testicular tissue.

Effects of Experimental Hyperthyroidism on Collagen IV and Laminin-α5 Gene Expression in Balb/C Mouse Seminiferous Tubules.

BACKGROUND: Hyperthyroidism is one of the disorders of the thyroid gland, an organ that controls the cellular and molecular behaviors of the seminiferous tubule basement membrane (BM), and ultimately, influences its evolutionary process. We aimed to investigate the effects of hyperthyroidism on immunohistochemical characteristics and gene expression levels of collagen IV and laminin-α5 in seminiferous tubules BM of Balb/C mice. MATERIALS AND METHODS: Twenty male Balb/C mice were divided into experimental and control groups. The experimental group received 500 mg/l of levothyroxine (L-thyroxine) diluted in drinking water for two months to inducing hyperthyroidism, which was confirmed by radioimmunoassay. At the end of the study, the mice were sacrificed, and their testes were extracted for immunohistochemistry and real-time polymerase chain reaction assays. RESULTS: Although a weak reaction was observed in the experimental specimens, no significant enhancement was noted in color intensity of type IV collagen in the seminiferous tubules BM of the experimental group as compared to the control group (P>0.05). Collagen IV gene expression results in the experimental group were not significantly different from the controls (P>0.05). Thus, there was a significant increase in laminin α5 gene expression compared to the control group (P=0.016). CONCLUSION: Considering the key role of collagen IV and laminin-α5 in the seminiferous tubule BM in the testes, the results of this study indicated that hyperthyroidism has important effects on both structures and functions of these two components.

Assessment of sperm morphology, chromatin integrity, and catSper genes expression in hypothyroid mice.

There is an evident relationship between the fertilizing capacity of sperm and the normal morphology, quality chromatin, and motility of sperm. It is well known that thyroid hormones are the important regulators of testicular function. A correlation was found between the hypothyroidism and sperm damages. The present study was conducted to investigate the effects of hypothyroidism on sperm morphology, chromatin quality, and motility. For this purpose, 20 male mice were divided into the control and the hypothyroid groups that received 0.05% 6-n-propyl-2-thiouracil (PTU) for 35 days. Sperm morphology with Papanicolaou staining and sperm chromatin quality with both Aniline Blue (AB) and Toluidine blue (TB) staining were assessed. Besides, immunohistochemistry and real-time PCR were performed to evaluate the changes of cation sperm channel (CatSper) genes. A significant increase in the sperm chromatin condensation was found in the hypothyroid mice compared to the control mice (p < 0.05). Furthermore, a significant decrease was observed in the morphology of normal sperm in hypothyroid mice compared to the controls (p < 0.05). The results showed that Hypothyroidism could downregulate the expression of CatSper genes. Immunohistochemical data confirmed the real time-PCR results. Furthermore, the results showed that hypothyroidism could adversely affect sperm morphology, sperm chromatin condensation, and CatSper gene expression in mice and these abnormalities may be related to the excessive production of reactive oxygen species (ROS) in a hypothyroid state.

Evaluation of oxidative stress indices after exposure to malathion and protective effects of ascorbic acid in ovarian tissue of adult female rats.

BACKGROUND: Malathion is one of organophosphate pesticides that is extensively used in farming and crops to control pests. Malathion induces oxidative stress in the various tissues such as the reproductive system. OBJECTIVE: To determine the effects of malathion on malondialdehyde (MDA) level and glutathione (GSH) content in female rat ovary tissue as well as to assess the protective role of Ascorbic Acid. METHODS: This study was carried out at the Department of Anatomy and Cell Biology (School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran) in 2015. In this experimental study, 30 adult, female, Wistar rats (weight range: 200-250 g) were divided into five groups, each group consisting of six rats: control group (no interventions), sham group (normal saline 0.9% 50 mg/kg), experimental group 1 (Ascorbic Acid 200 mg/kg), experimental group 2 (malathion 50 mg/kg), and experimental group 3 (malathion 50 mg/kg + Ascorbic Acid 200 mg/kg). Malathion, solvents and Ascorbic Acid were injected intraperitoneally. After two weeks, the animals were anaesthetized with intraperitoneal ketamine/xylazine (60 and 6 mg/kg, respectively) and then scarified, and the right ovarian was used to measure levels of MDA, a marker of lipid peroxidation, and GSH content. Data were analyzed by SPSS version 16, using descriptive statistics, One Way ANOVA, and Tukey-Kramer test. A p-value <0.05 was set as significance level. RESULTS: This study has shown that malathion increased MDA level and reduced GSH content compared with the control group (p<0.001). Also, administration of malathion in combination with Ascorbic Acid, reduced MDA level and increased the GSH content in rat ovarian tissue. CONCLUSION: Malathion induced lipid peroxidation and Oxidative stress in the ovarian of Rats. In addition, it appears that Ascorbic Acid, due to its antioxidant, can recover malathion-induced poisonous changes.

Evaluating the protective effects of vitamin C on serum and erythrocyte cholinesterase activity of male rats exposed to malathion.

INTRODUCTION: Malathion is one of organophosphate poisons (OPPs) that inhibit cholinesterase activity and induce oxidative stress in target organs, such as the reproductive system. The aim of this study was to assess the effects of Malathion on serum and erythrocyte cholinesterase activity in male rats and also to assess the protective effects of vitamin C in this regard. METHODS: This experimental study was performed in the Pharmacology Laboratory of the Pharmacy Faculty and in the Advanced Histology Techniques Laboratory of the Medical Faculty of Mashhad University of Medical Sciences (MUMS) in January 2014. Thirty male wistar rats, weighting 200-250 g, were divided into five groups of six. The different groups were exposed as follows: group 1: Malathion 50 mg/kg; group 2: Vitamin C; group 3: Malathion plus Vitamin C with the specified doses; sham group: normal saline; and control group: no exposure. After six weeks, 3 ml blood samples were taken from the rats, and titrimetric and Ellman methods were used to assess serum and erythrocyte cholinesterase activity, respectively. The data was analyzed by SPSS 16, and p < 0.05 was considered significant. RESULTS: The activities of serum and erythrocyte cholinesterase were inhibited significantly in the Malathion exposed group compared to the control group (p < 0.001). The administration of Vitamin C alone significantly increased the activities of serum and erythrocyte cholinesterase. The serum and erythrocyte cholinesterase inhibition showed improvement in the group that received both Malathion and Vitamin C. CONCLUSION: Malathion reduced the activities of serum and erythrocyte cholinesterase in exposed animals. It probably has the same intoxication effects on people who are exposed. Improvement of cholinesterase activity by antioxidant effects of Vitamin C suggests that Vitamin C supplementation can be used to decrease side effects of OPP exposure.

Neuronal cell reconstruction with umbilical cord blood cells in the brain hypoxia-ischemia.

BACKGROUND: Brain hypoxia-ischemia is a human neonatal injury that is considered a candidate for stem cell therapy. METHODS: 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 hypoxia-ischemia. 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. RESULTS: 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). CONCLUSION: The results show the potential of HUCB stem cells in reduction of neurologic deficits associated with neonatal hypoxia-ischemia.

Fibronectin regulation by vitamin C treatment in kidneys of nicotinic mice offspring.

BACKGROUND: Maternal cigarette smoking causes health risks and developmental defects in the offspring. So far, many studies have been conducted to suppress the effects of nicotine. However, the effects of coadministration of vitamin C and nicotine on extracellular matrix have not gained enough attention. OBJECTIVES: This study decided to investigate the effects of vitamin C on fibronectin expression in kidneys of mice offspring, treated with nicotine. MATERIALS AND METHODS: Eighteen female pregnant BALB/c mice were selected; six mice in the experimental group 1 (exp 1) received nicotine (3 mg/kg/day), six mice in the experimental group 2 (exp 2) received 3 mg/kg/day nicotine and 9 mg/kg/day vitamin C simultaneously, and six were used as the control group and received 3 mL/kg/day normal saline via intraperitoneal (IP) injection parallel to other groups, since the 6th day of gestation to the end of prenatal period. In the first days of delivery, fibronectin content of neonatal kidneys was studied by immunohistochemistry (IHC) assay and gene expression was studied by the real-time PCR. RESULTS: IHC results showed that fibronectin reaction significantly increased in proximal convoluted tubules of exp 1 compared with the control offspring; on the other hand, fibronectin reaction decreased in the mice offspring of exp 2. Gene expression results showed that fibronectin expression in the exp 1 offspring significantly increased compared with the control ones and fibronectin expression decreased in the mice offspring of exp 2. CONCLUSIONS: This study revealed that vitamin C could reduce the fibronectin accumulation effects of nicotine on kidney.

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

BACKGROUND: 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. OBJECTIVE: 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. MATERIALS AND METHODS: 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 1(st) experimental group were injected 3 mg/kg nicotine intrapritoneally from the 5(th) day of pregnancy to parturition daily and in 2(nd) experimental group the same procedure was repeated to the 10(th) 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. RESULTS: 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 2(nd) experimental group increased significantly compared to the 2(nd) control group (p=0.002). However, no significant difference was observed between the two experimental groups. CONCLUSION: 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.

Immunohistochemical study of type III collagen expression during pre and post-natal rat skin morphogenesis.

OBJECTIVE(S): Skin extracellular matrix, which contains type I and type III collagens, is involved in skin development. The aim of this study was to investigate type III collagen distribution pattern as well as its changes during pre and post-natal skin morphogenesis in rats. MATERIALS AND METHODS: Ventral skins of Wistar rat embryos at different stages from 10 to 20 gestational day (E10-E20) and also one month and one year post natal rat pups were fixed in normalin, embedded in paraffin and 5 µm thick sections were incubated with Anti type III collagen antibody. In order to detect staining intensity, the reactions were observed and graded by three examiners separately. Kruskal-Wallis non-parametric statistical test and SPSS software version 11.5 were used to compare differences between samples. RESULTS: Immunoreactivity of type III collagen was distributed weakly in the mesenchymal connective tissue on day 10 (E10). The observed reaction was increased onE12 and E14. This reaction was clear in basement membrane, relatively intensive in dermal papillae and moderate in dermal reticularis on E14. This immunoreactivity pattern was increased afterward on E16, not changed on E18 and decreased in dermal reticularis on E20. The density of collagen type III in dermal papillae and dermal reticularis in skin of one year old rats were decreased comparing to one month old rats. CONCLUSION: Our results showed that type III collagen is expressed and timely regulated during pre and post natal rat skin morphogenesis.