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.

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.

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.

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.

Exposure to mephedrone during gestation increases the risk of stillbirth and induces hippocampal neurotoxicity in mice offspring.

In recent years, abuse of synthetic cathinones, in particular, mephedrone, has increased among young adults worldwide. The study aim is to investigate the effects of mephedrone exposure during the gestational period on mice offspring outcomes, focusing on hippocampal neurotoxicity. The pregnant mice received mephedrone (50 mg/kg, sc) on a regular schedule (once daily on all days, from day 5 to 18 of gestation) or repeated schedule (thrice daily on day 5, 6, 11, 12, 17, and 18 of gestation) to simulate regular or recreational use of mephedrone, respectively. Results showed that the percentage of weight gain in pregnant mice was significantly lower in both regular and repeated schedule mephedrone groups (p < 0.01). Also, mephedrone significantly reduced the number and weight of delivered pups and increased the rate of stillbirth (p < 0.05). Immunohistochemistry and TUNEL assay showed an inhibition of cell proliferation (p < 0.05) and an increase of apoptosis (p < 0.05) in the hippocampus of delivered pups of the repeated schedule mephedrone group. This apoptotic effect was associated with enhanced expression of the pro-apoptotic Bax gene (p < 0.05) and reduction of expression of the anti-apoptotic Bcl-2 gene (p < 0.05) as evaluated by real-time PCR. The Morris water maze showed an impairment of spatial learning (p < 0.05) and reference memory (p < 0.01) in offspring born from litters exposed to mephedrone (repeated schedule). In conclusion, the present study has shown that regular and repeated exposure to mephedrone during the gestational period increases the risk of low birth weight and stillbirth. Also, repeated use of mephedrone impairs learning and memory processes through hippocampal damage.

Role of the Zeta Method in Intracytoplasmic Sperm Injection Outcomes in High Sperm DNA Fragmentation in Oligoasthenozoospermic Men.

BACKGROUND: New methods are needed to optimize intracytoplasmic sperm injection (ICSI) outcomes in oligoasthenozoospermic (OAS) men. We evaluated the level of DNA fragment index (DFI) in OAS men and its impact on ICSI outcomes. In addition, we used the zeta potential method for sperm selection to investigate the efficacy of this technique in improving ICSI outcomes. MATERIALS AND METHODS: This cross-sectional study was performed on 95 couples. Sperm parameters and sperm DNA fragmentation (SDF) were measured. The couples were divided into the following 3 groups: group I (n=30) where SDF was between 15% and 30%, and routine sperm was selected on the basis of motility and morphology; group II (n=34) where SDF was more than 30%, and the routine sperm selection method was applied on the basis of motility and morphology; and group III (n=31) where SDF was more than 30%, and the sperm selection was performed on the basis of the zeta method. The fertilization rate, embryo development, embryo quality, and implantation rate were evaluated in these 3 groups. RESULTS: The fertilization rate was significantly higher in group I compared with group II (P<0.05). The embryo development rate in group I was significantly higher than that in group II (P<0.001) and group III (P<0.05), and it was significantly lower in group II compared with group III (P<0.05). The embryo quality was higher in group III compared with group II (P<0.01). The implantation rate in group I was significantly higher than that in group II (P<0.05) and group III (P<0.05). CONCLUSIONS: The present study indicated that a higher level of SDF has an adverse effect on the ICSI outcome. Furthermore, the zeta potential technique can be a useful method for sperm selection in OAS men.

Effects of MDMA (ecstasy) on apoptosis and heat shock protein (HSP70) expression in adult rat testis.

BACKGROUND: This study was conducted to investigate the effects of MDMA (3,4-methylenedioxymethamphetamine, ecstasy) on apoptosis and heat shock protein expression in adult rat testis. METHODS: Twenty male rats were divided into four groups, two experimental groups (1 and 2), sham control and control. For 16 consecutive days, the experimental groups 1 and 2 were received 5 and 10 mg/kg intraperitoneal (ip) injection of ecstasy, respectively, and in the sham control group, the only saline was injected. In the control group there was no intervention. Finally, the rat's testes were removed and processed for terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and immunohistochemical techniques. RESULTS: Both doses of MDMA in experimental groups 1 and 2 significantly increased the mean number of TUNEL-positive cells in the germinal epithelium and Leydig cells (p < 0.05). Also in the experimental groups, the immunoreactivity of heat shock protein 70 (HSP70) significantly increased in the testis (p < 0.05). CONCLUSIONS: The findings revealed that MDMA administration increases the level of immunoreactivity of HSP70 and TUNEL-positive cells in the testicular tissue.

The effect of platelet-rich plasma on human mesenchymal stem cell-induced bone regeneration of canine alveolar defects with calcium phosphate-based scaffolds.

OBJECTIVES: Autologous bone transplantation known as the "gold standard" to reconstruction of osseous defects has known disadvantages. This study was designed to explore the effects of hydroxy-apatite/tricalcium-phosphate (HA/TCP) and platelet-rich plasma (PRP) on the osteogenesis ability of human adipose-derived mesenchymal stem cells (hAdMSCs) in vitro and in vivo. MATERIALS AND METHODS: hAdMSCs were incubated with HA/TCP granules and/or PRP in vitro and then, cell proliferation and differentiation was assessed by MTT assay, AZR S staining and SEM examination. In vivo, four cylindrical defects were drilled in the mandibular bones of 5 mongrel dogs and divided randomly into the following groups: I-autologous crushed bone, II- no filling material, III- HA/TCP and PRP, IV- PRP-enriched hAdMSCs seeded on HA/TCP granules. Inserted hAdMSCs were labeled to trace their contribution to bone tissue regeneration. Finally, cell tracing and tissue regeneration were evaluated by immunohistochemistry and histomorphometry methods, respectively. RESULTS: In vitro, co-incubation with HA/TCP granules significantly reduced proliferation and osteogenic differentiation ability of hAdMSCs; while PRP application promoted these capacities (P<0.05). In vivo, PRP-enriched hAdMSCs seeded on HA/TCP granules induced considerable bone formation in osseous defects (P<0.05). It was obviously shown that hAdMSCs were incorporated into the newly-formed bone. CONCLUSION: Based on this study, application of stem cells could offer a helpful therapeutic tool in bone tissue regeneration. Although inserted hAdMSCs were identifiable throughout the newly-formed bone tissue, their few number could be an indicator of indirect role of hAdMSCs in tissue regeneration.

Neuroprotective effects of Nigella sativa extract upon the hippocampus in PTU-induced hypothyroidism juvenile rats: A stereological study.

This study aimed to examine the neuroprotective effects of Nigella sativa (N. sativa) in the hippocampus of propylthiouracil (PTU)-induced hypothyroid rats during neonatal and juvenile growth. Twenty- five pregnant rats from early gestation (GD 0) were divided into five groups: (1) control (received drinking water), (2) PTU (received 0.005% PTU in drinking water), (3-5) PTU + NS 0.05%, PTU + NS 0.1%, PTU + NS 0.2% (along with PTU, received 0.05%, 0.1% and 0.2% W/V of N. sativa respectively) and treatment continued until postnatal day 60 (PN 60). The brains of male pups were removed for histological and stereological assessments. N. sativa extract significantly reduced the production of dark neurons and apoptotic cells in different areas of the hippocampus compared to the PTU group. Moreover, it significantly attenuated the effect of hypothyroidism on the volume reduction of the hippocampus. The results of the present study suggested that N. sativa extract has a potential ability to prevent the hippocampal neural damage after inducing hypothyroidism during neonatal and juvenile growth in rats.

Maternal exposure to titanium dioxide nanoparticles during pregnancy and lactation alters offspring hippocampal mRNA BAX and Bcl-2 levels, induces apoptosis and decreases neurogenesis.

INTRODUCTION: The usage of Titanium dioxide nanoparticles (TiO2-NPs) covers a vast area in different fields ranging from cosmetics and food to the production of drugs. Maternal exposure to TiO2-NPs during developmental period has been associated with hippocampal injury and with a decrease in learning and memory status of the offspring. However, little is known about its injury mechanism. This paper describes the in vivo neurotoxic effects of TiO2-NPs on rat offspring hippocampus during developmental period. MATERIAL AND METHODS: Pregnant and lactating Wistar rats received intragastric TiO2-NPs (100mg/kg body weight) daily from gestational day (GD) 2 to (GD) 21 and postnatal day (PD) 2 to (PD) 21 respectively. Animals in the control groups received an equal volume of distilled water via gavage. At the end of the treatment process, offspring were deeply anesthetized and sacrificed. Then brains of each group were collected and sections of the rat offspring's brains were stained using TUNEL staining (for detection of apoptotic cells) and immunostaining (for neurogenesis). Moreover, the right hippocampus (n=6 per each group) were removed from the right hemisphere for evaluating the expression of Bax and Bcl-2 level. RESULTS: Results of histopatological examination by TUNEL staining showed that maternal exposure to TiO2-NPs during pregnancy and lactation periods increased apoptotic cells significantly (P<0.01) in the offspring hippocampus. The immunolabeling of double cortin (DCX) protein as neurogenesis marker also showed that TiO2-NPs reduced neurogenesis in the hippocampus of the offspring (P<0.05). Moreover, in comparison with the control group, the mRNA levels of Bax and Bcl-2 in the TiO2-NPs group significantly increased and decreased, respectively (P<0.01). CONCLUSION: These findings provide strong evidence that maternal exposure to TiO2-NPs significantly impact hippocampal neurogenesis and apoptosis in the offspring. The potential impact of nanoparticle exposure for millions of pregnant mothers and their offspring across the world is potentially devastating.

Effects of Human Adipose-derived Stem Cells and Platelet-Rich Plasma on Healing Response of Canine Alveolar Surgical Bone Defects.

BACKGROUND: Due to the known disadvantages of autologous bone grafting, tissue engineering approaches have become an attractive method for ridge augmentation in dentistry. To the best of our knowledge, this is the first study conducted to evaluate the potential therapeutic capacity of PRP-assisted hADSCs seeded on HA/TCP granules on regenerative healing response of canine alveolar surgical bone defects. This could offer a great advantage to alternative approaches of bone tissue healing-induced therapies at clinically chair-side procedures. METHODS: Cylindrical through-and-through defects were drilled in the mandibular plate of 5 mongrel dogs and filled randomly as following: I- autologous crushed mandibular bone, II- no filling material, III- HA/TCP granules in combination with PRP, and IV- PRP-enriched hADSCs seeded on HA/TCP granules. After the completion of an 8-week period of healing, radiographic, histological and histomorphometrical analysis of osteocyte number, newly-formed vessels and marrow spaces were used for evaluation and comparison of the mentioned groups. Furthermore, the buccal side of mandibular alveolar bone of every individual animal was drilled as normal control samples (n=5). RESULTS: Our results revealed that hADSCs subcultured on HA/TCP granules in combination with PRP significantly promoted bone tissue regeneration as compared with those defects treated only with PRP and HA/TCP granules (P<0.05). CONCLUSION: In conclusion, our results indicated that application of PRP-assisted hADSCs could induce bone tissue regeneration in canine alveolar bone defects and thus, present a helpful alternative in bone tissue regeneration.

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.

Effect of drill-induced noise on hearing in non-operated ear.

BACKGROUND: The aim of this study was to evaluate the effect of drill-generated noise on hearing loss in non-operated ear and if any, was temporary or persistent. MATERIALS AND METHODS: In this prospective clinical study, 23 patients who had undergone mastoidectomy and normal contralateral hearing were enrolled. Patients were evaluated preoperatively and postoperatively (1 and 7 days) following surgery using low and high-frequency pure tone audiometry (PTA) and low and high-frequency transient evoked and distortion product otoacoustic emission (DPOAE) testing. RESULTS: Comparing preoperative and 1-day after surgery, PTA averages were significantly different at low frequencies, but no statistical significant differences were observed at 0.25 KHz and high-frequencies. Comparing 1-day after surgery and 7 days after surgery showed that, PTA averages at 0.5, 2 and 2 KHz were significantly different with no significant differences at the other average of thresholds in low and high frequencies; PTA average at 1 KHz was significantly different with, no significant differences at the other averages of thresholds in low and high frequencies. DPOAEs showed a significant difference preoperative and 1-day after surgery, 1-day and 7 days after, but DPOAEs were not significantly different. Transiently evoked otoacoustic emissions (TEOAEs) had a significant difference preoperative and 1-day after surgery, 1-day and 7 days after but when comparing preoperative and 7 days after surgery, TEOAEs were not significantly different. CONCLUSIONS: Drill-induced noise during ear surgery (mastoidectomy) can cause reversible changes in PTA, DPOAEs and TEOAEs in the non-operated ear.

Protective effects of ascorbic acid and garlic extract against lead-induced apoptosis in developing rat hippocampus.

Lead exposure has negative effects on developing nervous system and induces apoptosis in newly generated neurons. Natural antioxidants (i.e. Ascorbic acid and Garlic) might protect against lead-induced neuronal cell damage. The aim of the present study was to investigate the protective effects of Ascorbic acid and Garlic administration during pregnancy and lactation on lead-induced apoptosis in rat developing hippocampus. Timed pregnant Wistar rats were administrated with Lead (1500 ppm) via drinking water (Pb group) or lead plus Ascorbic acid (Pb + AA Group, 500 mg/kg, IP), or lead plus Garlic Extract (Pb + G Group, 1 ml garlic juice/100 g BW, via Gavage) from early gestation (GD 0) until postnatal day 50 (PN 50). At the end of experiments, the pups' brains were carefully dissected. To identify neuronal death, the brain sections were stained with TUNEL assay. Mean of blood and brain lead levels increased significantly in Pb group comparing to other studied groups (P < 0.01). There was significant reduction in blood and brain lead level in Pb + AA and Pb + G groups when compared to those of Pb group (P < 0.01). The mean number of TUNEL positive cells in the CA1, CA3, and DG was significantly lower in the groups treated by either Ascorbic acid or Garlic (P < 0.05). Administration of Ascorbic acid and Garlic during pregnancy and lactation protect against lead-induced neuronal cell apoptosis in the hippocampus of rat pups partially via the reduction of Pb concentration in the blood and in the brain.

The effects of exposure to titanium dioxide nanoparticles during lactation period on learning and memory of rat offspring.

Nanoscale titanium dioxide (TiO2), which is massively produced and widely used in living environment, seems to have a potential risk on human health. The central nervous system (CNS) is the potential susceptible target of nanoparticles, but the studies on this aspect are limited so far. The aim of this study was to evaluate the effects of exposure to TiO2 nanoparticles during lactation period on learning and memory of offspring. Lactating Wistar rats were exposed to TiO2 nanoparticles (100 mg/kg; gavage) for 21 days. The Morris water maze and passive avoidance tests showed that the exposure to TiO2 nanoparticles could significantly impair the memory and learning in the offspring. Therefore, the application of TiO2 nanoparticles and the effects of their exposure, especially during developmental period on human brain should be cautious.

DNA flow cytometric analysis in variable types of hydropic placentas.

BACKGROUND: Differential diagnosis between complete hydatidiform mole, partial hydatidiform mole and hydropic abortion, known as hydropic placentas is still a challenge for pathologists but it is very important for patient management. OBJECTIVE: We analyzed the nuclear DNA content of various types of hydropic placentas by flowcytometry. MATERIALS AND METHODS: DNA ploidy analysis was performed in 20 non-molar (hydropic and non-hydropic spontaneous abortions) and 20 molar (complete and partial moles), formalin-fixed, paraffin-embedded tissue samples by flow cytometry. The criteria for selection were based on the histopathologic diagnosis. RESULTS: Of 10 cases histologically diagnosed as complete hydatiform mole, 9 cases yielded diploid histograms, and 1 case was tetraploid. Of 10 partial hydatidiform moles, 8 were triploid and 2 were diploid. All of 20 cases diagnosed as spontaneous abortions (hydropic and non-hydropic) yielded diploid histograms. CONCLUSION: These findings signify the importance of the combined use of conventional histology and ploidy analysis in the differential diagnosis of complete hydatidiform mole, partial hydatidiform mole and hydropic abortion.

Glycoconjugates distribution during developing mouse spinal cord motor organizers.

BACKGROUND: The aim of this research was to study the distribution and changes of glycoconjugates particularly their terminal sugars by using lectin histochemistry during mouse spinal cord development. METHODS: Formalin-fixed sections of mouse embryo (10-16 fetal days) were processed for lectin histochemical method. In this study, two groups of horseradish peroxidase-labeled specific lectins were used: N-acetylgalactosamine, including Dolichos biflorus, Wisteria floribunda agglutinin (WFA), Vicia villosa, Glycine max as well as focuse-binding lectins, including tetragonolobus, Ulex europaeus, and Orange peel fungus (OFA). All sections were counterstained with alcian blue (pH 2.5). RESULTS: Our results showed that only WFA and OFA reacted strongly with the floor plate cells from early to late embryonic period of developing spinal cord. The strongest reactions were related to the 14, 15, and 16 days of tissue sections incubated with OFA and WFA lectins. CONCLUSION: The present study demonstrated that cellular and molecular differentiation of the spinal cord organizers is a wholly regulated process, and α-L-fucose, α-D-GalNAc, and α/ß-D-GalNAc terminal sugars play a significant role during the prenatal spinal cord development.

Histochemical study of retinal photoreceptors development during pre- and postnatal period and their association with retinal pigment epithelium.

OBJECTIVES: The aim of this study was to evaluate distribution and changes of glycoconjugates of retinal photoreceptors during both pre- and postnatal development. MATERIALS AND METHODS: Tissue sections from days 15 to 20 of Wistar rat embryos and 1 to 12 postnatal days of rat newborns including developing eye were prepared for lectinhistochemistry technique. Horseradish peroxidase (HRP)-labeled lectins including Vicia villosa (VVA), peanut agglutinin (PNA), Maclura pomifera (MPA) and wheat germ agglutinin (WGA-ІІ) were used. Alcian blue (pH 2.5) was used for counterstaining. RESULTS: Interphotoreceptor matrix (IPM) plays a crucial role in photoreceptors differentiation and acts as a mediator in interactions between photoreceptors and retinal pigment epithelium (RPE). Specific cell surface glycoconjugates secreted from cone cells could help us to distinguish these cells from rod photoreceptors. Our results for the first time revealed the strong reaction of cone photoreceptors with the cone-specific lectin (PNA) at postnatal day 12 (P12). Postnatal day 12 can be determined as the final differentiation of cone photoreceptors. CONCLUSION: According to our findings, we suggest that the generation of the eye photoreceptors begins from pre- natal period and their final differentiations will continue to postnatal period. Glycoconjugates including (ß-D-Gal [1-3]-D-GalNac) and (ß-D-Gal) terminal sugars play a critical role in the pre- and postnatal development and differentiation of retinal photoreceptors.