Thyroid hormone deficiency affects anxiety-related behaviors and expression of hippocampal glutamate transporters in male congenital hypothyroid rat offspring.

Thyroid hormones are crucial for brain development and their deficiency during fetal and postnatal periods can lead to mood and cognitive disorders. We aimed to examine the consequences of thyroid hormone deficiency on anxiety-related behaviors and protein expression of hippocampal glutamate transporters in congenital hypothyroid male offspring rats. Possible beneficial effects of treadmill exercise have also been examined. Congenital hypothyroidism was induced by adding propylthiouracil (PTU) to drinking water of pregnant Wistar rats from gestational day 6 until the end of the weaning period (postnatal day 28). Next, following 4 weeks of treadmill exercise (5 days per week), anxiety-related behaviors were examined using elevated plus maze (EPM) and light/dark box tests. Thereafter, protein expression of astrocytic (GLAST and GLT-1) and neuronal (EAAC1) glutamate transporters were measured in the hippocampus by immunoblotting. Hypothyroid rats showed decreased anxiety-like behavior, as measured by longer time spent in the open arms of the EPM and in the light area of the light/dark box, compared to control rats. Hypothyroid rats had significantly higher GLAST and GLT-1 and lower EAAC1 protein levels in the hippocampus than did the euthyroid rats. Following exercise, anxiety levels decreased in the euthyroid group while protein expression of EAAC1 increased and returned to normal levels in the hypothyroid group. Our findings indicate that thyroid hormone deficiency was associated with alterations in protein expression of glutamate transporters in the hippocampus. Up-regulation of hippocampal GLAST and GLT-1 could be at least one of the mechanisms associated with the anxiolytic effects of congenital hypothyroidism.

Bisphenol-S Influence on Oxidative Stress and Endocrine Biomarkers of Reproductive System: A Systematic Review and Meta-Analysis.

Background: Bisphenol-S (BPS), as a new human public health concern, was introduced to the plastic industry by BPA-free labeled products following the restrictions of Bisphenol-A (BPA) as a safe alternative. However, recent research has revealed a controversial issue. In this regard, the present study aimed to review the relationship between BPS exposure and reproductive system dis/malfunction. Methods: PubMed and other databases were searched up to January 2021. The standard mean difference (SMD) with a 95% confidence interval (CI) was calculated for the main parameters using the random-effects model. Finally, 12 studies with 420 subjects were included in this research. Forest plot, meta-regression, and non-linear dose-response effect were calculated for each parameter by random-effects model. Results: Based on the results of in vitro assessment, a significant increase was found in the oxidative stress parameters, including superoxide dismutase (SMD: 0.63, 95% CI: 0.321, 0.939), thiobarbituric acid reactive substances (SMD: 0.760, 95% CI: 0.423, 1.096), and reactive oxygen species (SMD: 0.484, 95% CI: 0.132, 0.835). In addition, the hormonal assessment revealed a significant decrease in male testosterone concertation (SMD: -0.476, 95% CI: -0.881, -0.071). Moreover, in vivo examination revealed a significant decrease in hormonal parameters, such as female testosterone (SMD: -0.808, 95% CI: -1.149, -0.467), female estrogen (SMD: -2.608, 95% CI: -4.588, -0.628), female luteinizing hormone (SMD: -0.386, 95% CI: -0.682, -0.089), and female follicle-stimulating hormone (FSH) (SMD: -0.418, 95% CI: -0.716, -0.119). Besides, linear and non-linear correlations were detected in the main parameters. Conclusion: In conclusion, based on the current meta-analysis, BPS was suggested to be toxic for the reproductive system, similar to the other bisphenols. Moreover, a possible correlation was indicated between oxidative and hormonal status disruption induced by BPS in male and female reproductive systems dis/malfunction.

Protective effects of treadmill exercise on apoptotic neuronal damage and astrocyte activation in ovariectomized and/or diabetic rat prefrontal cortex: molecular and histological aspects.

OBJECTIVE: Both estrogen deprivation and diabetes mellitus are known as risk factors for neuronal damage. Using an animal model of ovariectomized and/or streptozotocin (STZ)-induced diabetes mellitus, we examined expression of apoptosis-related proteins, neuronal damage, and astrocyte activation in prefrontal cortex of rats with/without treadmill exercise. METHODS: Adult female Wistar rats were divided into control, ovariectomized (Ovx, bilateral ovariectomy), diabetic (Dia, STZ 60 mg/kg; i.p.), and ovariectomized diabetic (Ovx + Dia) groups. Next, animals in each group were randomly subdivided into non-exercise and exercise subgroups. Animals in the exercise groups underwent moderate treadmill running for 4 weeks (5 days/week). Thereafter, expression of Bax, Bcl-2, and caspase-3, as apoptosis-related proteins, number of neurons, and number of glial fibrillary acidic protein (GFAP)-positive cells in prefrontal cortex were measured using immunoblotting, cresyl violet staining, and immunohistochemistry, respectively. RESULTS: In both Dia and Ovx + Dia groups, Bax and caspase-3 protein levels and number of GFAP-positive cells were higher than those in the control group, while Bcl-2 protein level and number of neurons compared were lower than the control group. Beneficial effects of exercise to prevent apoptosis-mediated neuronal damage and astrocyte activation were also observed in the Dia group. CONCLUSION: Based on our results, physical exercise could be beneficial to attenuate diabetes-induced neuronal damage in the prefrontal cortex via inhibition of apoptosis.

Neuroprotective Effects of Treadmill Exercise in Hippocampus of Ovariectomized and Diabetic Rats.

To determine detrimental effects of estrogen and insulin deficiencies on hippocampus, we examined apoptosis-induced neuronal damage and cholinergic system in ovariectomized and/or diabetic rat hippocampus. Possible neuroprotective effects of treadmill exercise were also investigated. Adult female Wistar rats were randomly divided into four groups (n = 5 rats/group) as follows: control, ovariectomized (Ovx), diabetic (Dia, streptozotocin (STZ) 60 mg/kg; i.p.), and Ovx + Dia groups. Each group was further subdivided into exercise and non-exercise groups. Animals in exercise groups were subjected to treadmill training, while those in non-exercise groups were placed on the stationary treadmill for 4 weeks (5 days/week). Apoptosis-related protein levels (i.e. Bax, Bcl-2, and caspase-3), number of survived neurons, and acetylcholinesterase (AChE) activity in the hippocampus were measured using Western blotting, Cresyl Violet staining, and Ellman assay, respectively. Both ovariectomy and diabetes increased expression of Bax and caspase-3 and decreased expression of Bcl-2 at protein levels. In addition, a significant decrease in the number of survived neurons was observed in both Ovx and Dia groups, while AChE activity was lower only in the Dia group. The Ovx + Dia group showed stronger apoptosis-induced neuropathology and inhibition of AChE activity. Treadmill exercise attenuated apoptosis-induced neuropathology in the Ovx and Dia groups and recovered AChE activity in the Dia group. Neuroprotective effects of treadmill exercise were mediated by inhibition of apoptosis. Moderate exercise protocol had no beneficial anti-apoptotic and neuroprotective effects in ovariectomized-diabetic rats.

Treatment of mouse cumulus-oocyte complexes with L-carnitine during vitrification and in vitro maturation affects maturation and embryonic developmental rate after parthenogenetic activation.

The technique of oocyte vitrification remains a challenge in most animal species. The present study aimed to evaluate the effects of cumulus cell presence and L-carnitine (LC) treatment during vitrification of selected immature oocytes by brilliant cresyl blue (BCB) staining on maturation and embryonic developmental rate after parthenogenetic activation. Immature oocytes were obtained from C57BL/6 female mice ovaries and stained with BCB. The BCB+ cumulus-oocyte complexes (COCs) were then selected and random parts of COCs were denuded from cumulus cells (denuded oocytes: DOs). COCs and DOs were treated with/out LC (0.6 mg/ml) during vitrification and in vitro maturation (IVM) procedures. A number of non-vitrified COCs were also treated with LC during the IVM process (fresh group). Maturation rate, intracellular glutathione (GSH) contents, and developmental competence of oocytes were also examined. The GSH levels in vitrified DOs+LC and vitrified COCs+LC groups were significantly higher (p < 0.01) than untreated vitrified-warmed COCs and DOs. Maturation rate and blastocyst developmental rate were reduced after the vitrification-warming procedure compared with the fresh group. The vitrified COCs+LC group showed a higher percentage of mature oocytes and the ability to develop to blastocyst stage than the vitrified-warmed DOs group (p < 0.01). These data indicated that the presence of cumulus cells around the competent oocyte and LC treatment during vitrification and IVM procedure could improve parthenogenetic developmental competence of vitrified-warmed oocytes by increasing GSH levels and accelerating oocyte maturation.

Apoptosis is involved in paraoxon-induced histological changes in rat cerebellum.

Acute toxicity of organophosphorus compounds is primarily caused by inhibition of acetylcholinesterase (AChE) at cholinergic synapses. The current study was designed to investigate the effects of paraoxon on histological changes as well as the role of mitochondrion-dependent apoptosis in causing this damage in the rat cerebellum. Adult male Wistar rats were intraperitoneally injected with paraoxon at 0.3, 0.7, or 1 mg/kg. Control animals were injected with corn oil as a vehicle. At 14 or 28 days after intoxication, histological changes and alterations in the expression of apoptosis-related proteins, including Bax, Bcl-2, and caspase-3, were investigated in the cerebellum using cresyl violet staining and western blotting, respectively. Findings showed the decreased thickness of both molecular and granular layers and reduction in the number of Purkinje cells in animals treated with a higher convulsive dose of paraoxon (1 mg/kg). In addition, exposure of rats to 1 mg/kg of paraoxon activated apoptosis pathway confirmed by an increase in Bax and caspase-3 and a decrease in Bcl-2 protein levels. According to our results, cerebellar histological changes and alterations in the expression of apoptosis-related proteins occur following exposure to a high convulsive dose of paraoxon and persist for a long time.

Relationship between metabolic syndrome and pulmonary function in workers with respiratory dust exposure in Iran.

AIMS: we designed a study to investigate the relationship between metabolic syndrome (MetS) and lung function in workers with dust exposure based on five years of longitudinal study data. METHODS: In this historical cohort study that conducted in iron ore mine, non-smoker male workers who exposed to dust, were enrolled. MetS was determined according to the National Cholesterol Education Program Adult Treatment Panel III. New spirometry parameters and spirometry from 5 years ago, were compared. RESULTS: In this study 192 workers were identified without MetS and 77 with MetS. The mean of all lung parameters was lower in subjects with MetS, but it was not statistically significant. The median decline in FEV1 and FVC in 5 years was greater in subjects with MetS but were only significant for a decline in FEV1 (P-Value = 0.04). Linear regression analysis showed a significant relationship between a decline in FEV1 and waist circumference (P-Value = 0.001) when adjusted for age, BMI, physical activity level. CONCLUSION: In this study, a significant association between mean decline in FEV1 in 5 years and MetS in dust-exposed workers was demonstrated. Decline in FEV1 in 5 years was significantly associated with a Waist circumference as one of the components of MetS.

The Protective Effects of L-Carnitine and Zinc Oxide Nanoparticles Against Diabetic Injury on Sex Steroid Hormones Levels, Oxidative Stress, and Ovarian Histopathological Changes in Rat.

Diabetes mellitus is a common chronic metabolic disorder. This study aimed to investigate the effects of co-treatment with L-carnitine (LC) and zinc oxide nanoparticles (ZnONPs) on serum levels of sex hormones, oxidative stress, and ovarian histopathology in streptozotocin (STZ)-induced diabetic rats. Female Wistar rats (n = 56, 180-220 g) received a single intraperitoneal (IP) injection of STZ (65 mg/kg). They were randomly assigned into the following groups: diabetic group (Dia), Dia+Met group (100 mg metformin/kg/day), Dia+LC group (200 mg/kg/day), Dia+ZnONPs group (10 mg/kg/day), and Dia+LC+ZnONPs group (200 mg LC/kg/day and 10 mg ZnONPs/kg/day). Control group (Ctl) received the same volume of STZ solvent. After 21 days of treatment, blood serum was centrifuged for sex hormone assays. The right ovary was used for biochemical analysis, and the left ovary was fixed in 10% neutral buffered formalin for histological assessment. The levels of estradiol, progesterone, FSH, and LH significantly increased in the Dia+ZnONPs+LC group (P < 0.001) compared with the Dia group. Co-treatment with LC and ZnONPs reduced malondialdehyde and carbonyl protein and increased glutathione, catalase, and superoxide dismutase activities in ovarian tissue compared with the Dia group (P < 0.05). Moreover, the number of all ovarian follicles significantly increased in this group compared with the Dia group (P < 0.05). The results of this study indicated that co-treatment with LC and ZnONPs could preserve ovarian function by increasing sex hormones levels and antioxidant activity and decreasing lipid peroxidation in diabetic rats. Therefore, this compound supplementation may improve ovulation and fertility in people with diabetes mellitus.

Effects of treadmill exercise on cognitive functions and anxiety-related behaviors in ovariectomized diabetic rats.

Diabetes mellitus and ovarian hormone deficiency are associated with mood and cognition disorders. We aimed to study the possible beneficial effects of treadmill exercise on cognitive impairments and anxiety-related behaviors in ovariectomized diabetic rats. Fourteen days after bilateral ovariectomy or sham operation, adult female Wistar rats (n = 7 per group) received an intraperitoneal injection of streptozotocin (60 mg/kg) for induction of diabetes mellitus or citrate buffer. After 14 days, the animals were subjected to treadmill running or placed on the stationary treadmill for 4 weeks (5 days/week). The animals were then subjected to Morris water maze (MWM), elevated plus maze (EPM), and open field test to determine spatial learning and memory, anxiety-related behaviors, and locomotor activity, respectively. Spatial learning decreased in diabetic and ovariectomized + diabetic (Ovx + Dia) groups. Decreased spatial memory and increased anxiety levels were observed in the Ovx, Dia, and Ovx + Dia groups. Animals in the Ovx + Dia group showed reduced locomotor activity. Treadmill exercise improved spatial memory and anxiety levels in the Ovx and Dia groups, while had no significant effect on spatial learning and memory, anxiety levels, and locomotor activity in the Ovx + Dia group. Collectively, our results showed that physical exercise has beneficial effects on anxiety-related behaviors and spatial memory, but not of learning, in the ovariectomized and diabetic animals. Co-existence of ovariectomy and diabetes exacerbates anxiety-related behaviors, but not cognitive functions, compared to diabetes or ovariectomy alone; these complications could not be improved by exercise.

Paraoxon-induced damage in rat hippocampus is associated with alterations in the expression of apoptosis-related proteins.

To determine the possible role of apoptosis in the development of paraoxon-induced brain damage, we evaluated expression of apoptosis-related proteins, the extent of neuronal damage, and activation of astrocytes in rat hippocampus. Adult male Wistar rats were intraperitoneally injected with one of three doses of paraoxon (0.3, 0.7, or 1 mg/kg) or corn oil (vehicle). After 14 or 28 days, expression of apoptosis-related proteins, including B-cell leukemia/lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), and caspase-3, as well as the number of neurons and glial fibrillary acidic protein (GFAP) positive cells in hippocampus were examined by western blot, cresyl blue staining, and immunohistochemistry, respectively. After 14 and 28 days, Bax and caspase-3 proteins were significantly increased in rats receiving 0.7 and 1 mg/kg of paraoxon. A significant decrease in Bcl-2 protein levels was also observed in 0.7 and 1 mg/kg groups after 14 days and in 1 mg/kg group after 28 days. Animals treated with 1 mg/kg of paraoxon showed a significant decrease in the number of neurons in the CA1 area. Also, those treated with 0.7 and 1 mg/kg of paraoxon showed an increase in the number of GFAP positive cells in both CA1 and CA3 areas as well as a significant decrease in survived neurons in the CA3 area. Our results indicated that neuronal damage induced by convulsive doses of paraoxon in rat hippocampus is mediated in part through apoptosis mechanism. Activation of astrocytes might lead to reduced extent of damage and damage and consequently increased neuronal survival.

Acrosome and chromatin integrity, oxidative stress, and expression of apoptosis-related genes in cryopreserved mouse epididymal spermatozoa treated with L-Carnitine.

Oxidative stress is believed to be an important cause of sperm damage during freezing. l-Carnitine (LC) may have the potential to improve sperm quality after frozen-thawed process. The present study aimed to investigate the effect of LC supplementation in cryoprotectant media of mouse epididymal sperm on post-thaw sperm quality and expression of apoptosis-related genes. Male BALB/cJ mice spermatozoa were cryopreserved in a cryoprotectant medium containing 2.5 or 5 mM LC. The untreated group was cryopreserved with the cryoprotectant medium only. Six months following cryopreservation, the samples were thawed and sperm quality parameters, chromatin and acrosome integrity, reactive oxygen species (ROS) and glutathione (GSH) levels, mitochondrial activity, and mRNA expression of Bax and Bcl-2 were assessed. The results demonstrated that the concentration of 5 mM LC in cryoprotectant media exhibited higher values for the sperm quality parameters and integrity of chromatin and acrosome in post-thaw spermatozoa than those of the untreated group. Furthermore, sperm ROS levels decreased while GSH and mitochondrial activity levels increased in 5 mM LC group compared to those in the untreated group (P < 0.01). In 5 mM LC-treated group, Bax was down-regulated (P < 0.05) while Bcl-2 was up-regulated (P < 0.001) compared to the untreated group. Collectively, LC supplementation of cryoprotectant medium improved the quality of frozen-thawed mouse epididymal spermatozoa, as showed reduced ROS level and Bax expression as well as increased GSH, mitochondrial activity, and Bcl-2 expression.

Effects of Paraoxon Exposure on Expression of Apoptosis-Related Genes, Neuronal Survival, and Astrocyte Activation in Rat Prefrontal Cortex.

Paraoxon is the bioactive metabolite of organophosphate (OP) pesticide, parathion. This study aimed to evaluate the expression of apoptosis-related genes and histopathological changes in rat prefrontal cortex following exposure to three different doses of paraoxon. Paraoxon (0.3, 0.7, or 1 mg/kg) or corn oil (vehicle) were intraperitoneally injected to adult male Wistar rats. After 14 or 28 days, mRNA and protein levels of Bax, Bcl-2, and caspase-3 were measured in prefrontal cortex using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blotting, respectively. In addition, neuronal injury and astrocyte activation were assessed using cresyl violet staining and glial fibrillary acidic protein (GFAP) immune-positive cells, respectively. Treatment with 0.7 and 1 mg/kg of paraoxon increased mRNA and protein levels of Bax and caspase-3 at 14 and 28 days post-exposure, while mRNA and protein levels of Bcl-2 decreased only in 1 mg/kg group after 14 days. Furthermore, a significant decrease in the number of neurons and a significant increase in the number of GFAP-positive cells were observed in rats receiving 0.7 and 1 mg/kg of paraoxon at both time points. Collectively, our results showed that apoptosis is a major mechanism for neuronal damage after exposure to paraoxon. Also, paraoxon-induced neuronal loss was correlated with activation of astrocytes. Since paraoxon-induced neuronal damage is closely related to convulsion, clinical management of convulsion could protect neuronal brain damage.

Comparison of Epigenetic Modifier Genes in Bovine Adipose Tissue-Derived Stem Cell Based Embryos, as Donors, with In Vitro and Parthenogenesis Embryos.

OBJECTIVE: Regarding that undifferentiated mesenchymal stem cells, as donor cells, require less epigenetic reprogramming, possibility of using bovine adipose tissue-derived stem cells (BASCs) with low level of DNMTs and HDACs expression was evaluated. MATERIALS AND METHODS: In this experimental study, we examined gene expression of epigenetic modifiers including DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) and histone deacetylases (HDAC1-3), as well as protein levels of histone H3 acetylation at lysine 9 (H3K9ac) and POU5F1 (also known as OCT4) at two stages of preimplantation development among in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) groups. RESULTS: The results revealed that developmental competence of IVF embryos was higher than SCNT embryos (P<0.05). In the PA and SCNT groups, DNMT1, HDAC2 and HDAC3 mRNA were overexpressed (P<0.05), and proteins levels of H3K9ac and POU5F1 were reduced at 6-8 cells and blastocyst stages compared to IVF (P<0.05). The mRNA expression of DNMT1 and HDAC1 and proteins levels of POU5F1 and H3K9ac were significantly different between SCNT and PA groups (P<0.05) in both developmental stages (except HDAC1 in blastocyst stage). CONCLUSION: The SCNT embryos derived from BASCs have endured considerable nuclear reprogramming during early embryo development. Comparison of PA and SCNT blastocysts demonstrated that HDAC1 and DNMT1 may attribute to developmental competence variability of bovine embryos.

Effects of L-Carnitine on the sperm parameters disorders, apoptosis of spermatogenic cells and testis histopathology in diabetic Rats.

BACKGROUND: Diabetes mellitus affects male reproductive system that is known to cause male infertility. OBJECTIVE: The aim of the present study was to assess the effects of L-carnitine (LC) on sperm parameters, apoptosis of spermatogenic cells and testis histopathology in Streptozotocin-induced diabetic Rats. MATERIALS AND METHODS: The study was carried out on 36 male Wistar adult rats (220 ± 30 gr) randomly divided into six groups (n = 6/each). 1 (Control); 2 (LC 100 mg/kg); 3 (Diabetic); 4, 5, and 6 (Diabetic + LC 50 or 100 or 200 mg/kg, respectively). Daily injections were administered intraperitoneally for 48 days. Then, rats were sacrificed, left testis and epididymis were harvested for sperm analysis and histopathology, morphometric and spermatogenesis assessments, and Tunnel assay. RESULTS: L-carnitine in group 6 significantly decreased blood glucose level (p < 0.01) in comparison with group 3. L-carnitine in groups 5 and 6 significantly (p < 0.001) and dose-dependently increased the count, motility, viability, maturity, and chromatin quality of sperm and decreased the abnormal morphology of sperm in comparison with group 3. In groups 4, 5, and particularly 6, in comparison with group 3, there has been a significant difference in the increase of seminiferous tubule diameter, germinal epithelium height (p < 0.001), maturity quality of the seminiferous tubules (p < 0.001), decrease apoptosis of spermatogenic cells (p < 0.001), and testis tissue histopathological complications. CONCLUSION: The data obtained from the present study suggest that in the diabetic rats, LC decreases serum glucose level, improves the diameter and thickness of the epithelium of spermatogenic cells, reduces germ cells' apoptosis, and improves epididymal sperm parameters. Therefore, it seems that LC plays an effective role in diabetes-induced infertility.

Anxiolytic activity of paraoxon is associated with alterations in rat brain glutamatergic system.

Exposure to organophosphate (OP) compounds leads to behavioral alterations. To determine whether paraoxon has effects on anxiety, anxiety-like behaviors were assessed in paraoxon-exposed rats. Protein expression of glutamate transporters has also been measured in hippocampus and prefrontal cortex. Three doses of paraoxon (0.3, 0.7, or 1 mg/kg) or corn oil (vehicle) were intraperitoneally injected to adult male rats. At 14 or 28 days after exposure, behavioral tests were done using elevated plus-maze (EPM) or open field tests. Thereafter, animals were sacrificed and both hippocampi and prefrontal cortices were extracted for cholinesterase assay and western blotting. Animals treated with convulsive doses of paraoxon (0.7 and 1 mg/kg) showed an increase in percentage of time spent in open arms and percentage of open arm entries in the EPM. In the open field test, an increase in the time spent in central area was observed in rats treated with the same doses of paraoxon. These effects of paraoxon were independent of any changes in locomotor activity. There was an increase in both astrocytic glutamate transporter proteins (GLAST and GLT-1) in the hippocampus of animals treated with 0.7 and 1 mg/kg of paraoxon. In the prefrontal cortex, protein levels of the GLAST and GLT-1 increased in 0.7 and decreased in 1 mg/kg groups. Only a significant decrease in EAAC1 protein was observed in the prefrontal cortex at 14 days following exposure to 1 mg/kg of paraoxon. Collectively, this study showed that exposure to convulsive doses of paraoxon induced anxiolytic-like behaviors in both behavioral tests. This effect may be attributed to alterations of glutamate transporter proteins in the rat hippocampus and prefrontal cortex.

Neuroprotective effects of hyperbaric oxygen (HBO) therapy on neuronal death induced by sciatic nerve transection in rat.

BACKGROUND: Recent studies shows that hyperbaric oxygen (HBO) therapy exerts some protective effects against neural injuries. The purpose of this study was to determine the neuroprotective effects of HBO following sciatic nerve transection (SNT). METHODS: Rats were randomly divided into five groups (n = 14 per group): Sham-operated (SH) group, SH + HBO group, SNT group, and SNT + pre- and SNT + post-HBO groups (100% oxygen at 2.0 atm absolute, 60 min/day for five consecutive days beginning on 1 day before and immediately after nerve transaction, respectively). Spinal cord segments of the sciatic nerve and related dorsal root ganglions (DRGs) were removed 4 weeks after nerve transection for biochemical assessment of malodialdehyde (MDA) levels in spinal cord, biochemical assessment of superoxide dismutase (SOD) and catalse (CAT) activities in spinal cord, immunohistochemistry of caspase-3, cyclooxigenase-2 (COX-2), S100beta (S100ß), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) in spinal cord and DRG. RESULTS: The results revealed that MDA levels were significantly decreased in the SNT + pre-HBO group, while SOD and CAT activities were significantly increased in SNT + pre- and SNT + post-HBO treated rats. Attenuated caspase-3 and COX-2 expression, and TUNEL reaction could be significantly detected in the HBO-treated rats after nerve transection. Also, HBO significantly increased S100ß expression. CONCLUSIONS: Based on these results, we can conclude that pre- and post-HBO therapy had neuroprotective effects against sciatic nerve transection-induced degeneration.

Differential expression of glutamate transporters in cerebral cortex of paraoxon-treated rats.

Glutamatergic system is involved in pathological effects of organophosphorus (OP) compounds. We aimed to determine in vivo effects of paraoxon, the bioactive metabolite of parathion, on the expression of glutamate transporters as well as Bax and Bcl2 in rat cerebral cortex. Male Wistar rats received an intraperitoneal (i.p.) injection of one of three doses of paraoxon (0.3, 0.7, or 1mg/kg) or corn oil as vehicle (1ml/kg). After 4 or 18h, cerebral cortices were dissected out and used for quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blot assays to measure mRNA and protein levels, respectively. The cortical glial glutamate transporters (GLAST and GLT-1) were up-regulated in animals treated with 0.7mg/kg of paraoxon, but down-regulated in 1mg/kg group. Neuronal glutamate transporter (EAAC1) was unchanged in 0.7mg/kg treated rats, while reduced in 1mg/kg group. No significant difference was found in the mRNA and protein expression of EAAC1 in animals intoxicated with 0.3mg/kg of paraoxon. Paraoxon (1mg/kg) resulted in an up-regulation of Bax and down-regulation of Bcl2 mRNA levels in the rat cerebral cortex. These results indicate that paraoxon can differentially regulate expression of glutamate transporters at mRNA and protein levels in the cerebral cortex. Changes in the expression of glutamate transporters are closely related to paraoxon-induced seizure activity.

Supplementation of L-carnitine during in vitro maturation of mouse oocytes affects expression of genes involved in oocyte and embryo competence: An experimental study.

BACKGROUND: Oocyte developmental competence is one of the key factors for determining the success rate of assisted reproductive technique. OBJECTIVE: The aim of the current study was to investigate the effect of L-carnitine (LC) supplementation during in vitro maturation (IVM), on preimplantation embryo development and expression of genes involved in embryo competence derived from oocytes selected with brilliant cresyl blue (BCB) test. MATERIALS AND METHODS: Cumulus-oocyte complexes (COCs) were obtained from NMRI mice ovaries. COCs were stained with BCB and then BCB+ (colored cytoplasm) oocytes cultured in IVM medium supplemented with 0.3 or 0.6 mg/ml LC. COCs untreated with LC were used as control. Fertilization rate and blastocyst development rate were determined after in vitro fertilization. In addition, quantitative reverse transcriptase polymerase chain reaction was used to measure relative genes expression related with development (Ccnb1, Mos, Ces5, and Dppa2) and apoptosis (Bax and Bcl-xL) in oocytes and embryos. RESULTS: Oocytes treated with both LC concentrations showed higher blastocyst development rate compared with untreated oocytes (p<0.01). Moreover, fertilization rate was increased in oocytes treated with 0.6 mg/ml LC (p<0.01). Treatment of oocytes with both LC concentrations increased (p<0.01) the level of Ccnb1 mRNA in MII oocytes. The two-cell stage embryos and blastocysts derived from LC-treated oocytes (0.6 mg/ml) showed increased the expression levels of Dppa2 and Bcl-xl mRNA, respectively (p<0.01). CONCLUSION: The results of the present study show that adding of LC to the IVM medium of BCB+ oocytes can ameliorate reproductive success following in vitro fertilization.

Alterations in mRNA and protein expression of glutamate transporters in rat hippocampus after paraoxon exposure.

Organophosphates affect brain function through a variety of mechanisms beyond their shared role as cholinesterase inhibitors. The aim of the current study was to investigate the changes in the expression of glial (GLAST and GLT-1) and neuronal (EAAC1) glutamate transporters at mRNA and protein levels in paraoxon-treated rat hippocampus. Adult male Wistar rats were intraperitoneally treated with either vehicle (corn oil) or one of three dosages of paraoxon (0.3, 0.7 or 1mg/kg). After 4 or 18h, both hippocampi of each rat were collected to detect mRNA and protein expression of glutamate transporters using the quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blotting, respectively. Animals treated with 0.3mg/kg paraoxon showed no difference in mRNA and protein levels of the glutamate transporters when compared with control group. At 4h after exposure with 0.7 and 1mg/kg paraoxon, the expression of GLAST and GLT-1 increased at mRNA and protein levels and remained elevated after 18h. No difference in the expression of EAAC1 at mRNA and protein levels was observed in any paraoxon-treated groups compared with the control group. This study showed an increased expression of glial (GLAST and GLT-1), but not neuronal (EAAC1) glutamate transporters, in adult rat hippocampus following administration of convulsive dosages of paraoxon. These suggest a protective and compensatory adaptation for effective uptake of glutamate in hippocampus induced by paraoxon and thus attenuating seizure activity.

Effect of L-carnitine supplementation on maturation and early embryo development of immature mouse oocytes selected by brilliant cresyle blue staining.

PURPOSE: The present study was designed to investigate the effect of L-carnitine treatment during IVM on nuclear and cytoplasmic maturation of immature oocytes selected by Brilliant Cresyle Blue (BCB) staining, and their subsequent developmental competence. MATERIALS & METHODS: Compact cumulus-oocyte complexes (COCs) were collected from NMRI mice ovaries and stained with BCB staining. BCB+ (colored cytoplasm) oocytes were then cultured in tissue culture medium (TCM) 199 with 0.0, 0.3 and 0.6 mg/ml L-carnitine. RESULTS: The both L-carnitine concentrations significantly increased the intracellular glutathione (P<0.001), nuclear maturation (P<0.01) and expression levels of cyclin-dependent kinase1 (CDK1) (P<0.05). Moreover, treated oocytes with 0.6 mg/ml L-carnitine showed increased (P < 0.05) expression of mitogen-activated protein kinase1 (MAPK1) mRNA. Also, adding L-carnitine (0.6 mg/ml) to IVM medium significantly increased the cleavage rate (P<0.05). The blastocyst development rate (BDR) in the both L-carnitine treated groups was significantly higher (P<0.001) than the control group. L-carnitine had no significant effect on total blastocyst cell numbers. CONCLUSIONS: These data indicated that L-carnitine supplementation during IVM of immature BCB+ oocytes improved preimplantation developmental competence of oocytes after IVF, probably by accelerating cytoplasmic and nuclear maturation of oocytes. It may provide a novel approach to improving ART outcomes in infertile couples.