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

Effects of essential and non-essential amino acids on in-vitro maturation, fertilization and development of immature bovine oocytes

Background: Addition of amino acids to the culture medium is beneficial for embryonic development in many species. Objective: The objective of this study was to investigate the effects of amino acids on the in vitro maturation and embryonic development of the bovine oocyte. Materials and Methods: Bovine ovaries were collected from a local abattoir and brought into laboratory. Cumulus-oocyte complexes (COCs; n=1212) were aspirated from follicles (2-8 mm in diameter) and randomly assigned to four groups for maturation in culture: (1) Basic medium alone as control; (2) Basic medium supplemented with 2% MEM essential amino acids solution; (3) Basic medium supplemented with 1% MEM non-essential amino acids solution; and (4) Basic medium supplemented with 2% MEM essential amino acids solution + 1% MEM non-essential amino acids solution. COCs were incubated in 1 ml maturation medium in an Organ culture dish at 38.5°C in an atmosphere of 5% CO2 with high humidity. After 24 h of culture, 372 oocytes were fixed to determine maturation rate and the remaining oocytes were used for in vitro fertilization (IVF). Following 18 h of insemination, 437 oocytes were fixed and examined for fertilization and 403 oocytes were further cultured. Results:There were no differences in maturation rates and penetration rates among the four groups. Although oocyte cleavage rates were not different in the four groups, embryo development up to the 8-cell stage and blastocyst were significantly higher (p<0.05) in Group (2) and (4) than in the Control and Group (3). Conclusion: These results indicate that the presence of amino acids, especially essential amino acids in the maturation medium is beneficial to oocyte cytoplasmic maturation and subsequent early embryo development in vitro

Combined protective effect of zinc oxide nanoparticles and melatonin on cyclophosphamide-induced toxicity in testicular histology and sperm parameters in adult Wistar rats

Background: Cyclophosphamide (CP) has been known as an anticancer drug with several side effects on various organs such as a male reproductive system that can cause infertility. Objective: To evaluate the possible combined effects of zinc oxide nanoparticles (nZno) and melatonin (Mel) on sperm parameters and histopathological changes of the testis in CP-treated rats. Materials and Methods: 42 adult male Wistar rats were divided into six groups. GI: control, GII: 60 mg/kg/wk CP, GIII and GIV, 10 mg/kg/wk Mel and 5mg/kg/wk nZno and GV: 5 mg/kg/wk nZno and 10 mg/kg/wk Mel were given 2 hr prior to CP injection, respectively,GVI: 5mg/kg/wk nZno and 10 mg/kg/wk Mel simultaneously. After 8 wk of treatment, rats were sacrificed and testis and epididymis were harvested for further evaluation. Results: The CP-treated group showed significant decreases in the body, testes and epididymis weights and sperm parameters (sperm count, viability, motility) with an increase abnormal sperms when compared with the control (p<0.001), as well as many histological alterations included decreased diameters of seminiferous tubules and Johnsen&rsquo;s Testicular Score (with degeneration, desquamation, multi-nucleated giant cell formation), whereas combined treatment (GV), showed more protective effects on CP-induced reproductive system damage compared with groups III or IV (p<0.001). Conclusion: These results suggest simultaneous administration of Mel and nZno have more effectively protections against CP-induced reproductive damage than Mel or nZno alone

Expression of Spermatogonial and Pluripotency Markers in Spermatogonial Stem Cells after Treatment with Different Culture Factors

Background: As condition and component of culture determine fate map of spermatogonial stem cells (SSCs), the aim of this study was to evaluate of growth factors GDNF, LIF and RA on proliferation and differentiation of SSC. Materials and Methods: SSCs were cultured in two groups: The first group GDNF and LIF and the second group RA. The number of clumps and colony formation was monitored during 1 month in culture. To identification of the colony, stained with PLZF using immunostaining. Pluripotency gene Oct 4 and neural markers MAP2, NeuroD and Nestin were analyzed by RT-PCR.  Results: In the presence of GDNF and LIF, cells proliferated rapidly and many compact clumps were appeared whereas after exposure to RA cells formed small clumps. The results of immunocytochemistry shows PLZF was detected in the group GDNF & LIF. RT-PCR showed high level expression Oct 4 in the group GDNF and LIF whereas neural markers MAP2, NeuroD and Nestin were expressed in the group RA. Conclusions: GDNF and LIF are essential for self-renewal and colony formation of SSCs that confirm the stem cells activity of these cells but RA inhibits stem cell activity of SSCs and induces neural differentiation of these

Antimicrobial peptides-loaded smart chitosan hydrogel: Release behavior and antibacterial potential against antibiotic resistant clinical isolates

In this study, we synthesized thermo -responsive chitosan (TCTS) hydrogels, and loaded with different concentrations of antimicrobial peptide (AMP) (0, 4, 8 and 16 µg.ml - 1 ) to fabricate an antibacterial wound dressing against resistant clinical isolates. Physico -chemical properties, release behavior, cytobiocompatibility and antibacterial activity of the AMP -TCTS hydrogels against standard strain and resistant Acinetobacter baumannii were fully determined in vitro. The TCTS -40% β -glycerolphosphate hydrogels showed a gelation time of 15 min at 37 °C. 80% weight loss at day 35 with no changes in pH value was observed. AMP -TCTS hydrogels showed a burst release of AMP (around 40%) at day 1, and a controlled release up to day 7. A dramatic water uptake was observed at first 4 h, and then continued for 10 h in a steady manner. All the AMP -TCTS hydrogels showed excellent cytobiocompatibility for human fibroblasts. The TCTS showed no antibacterial activity against both standard strain and clinical isolates. All the AMP - TCTS hydrogels had strong antibacterial activity against standard strains, but only 16 µg.ml - 1 showed antibacterial behavior against resistant A. baumannii . Our results strongly suggest the 16 µg.ml - 1 AMP -TCTS hydrogel as a n excellent antibacterial wound dressing against resistant A. baumannii , and now promises to proceed with pre -clinical investigations

Trans-Differentiation of Human Dental Pulp Stem Cells Into Cholinergic-‎Like Neurons Via Nerve Growth Factor

Introduction: Cell therapy has been widely considered as a therapeutic approach for neurodegenerative diseases and nervous system damage. Cholinergic neurons as one of the most important neurons that play a significant role in controlling emotions, mobility, and autonomic systems. In this study, human dental pulp stem cells (hDPSCs) were differentiated into the cholinergic neurons by β-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase.  Methods: The hDPSCs were evaluated for CD73, CD31, CD34, and Oct-4. Concentration-time relationships for NGF were assessed by evaluating the viability rate of cells and the immune response to nestin, neurofilament 160, microtubule-associated protein-2, and choline acetyltransferase. Results: The hDPSCs had a negative response to CD34 and CD31. The optimal dose for the NGF was 50 ng/mL seven days after the induction when the highest percentage of expressing markers for the cholinergic neurons (ChAT) was detected. Conclusion: The results of this study provided a method for producing cholinergic neurons by hDPSCs, which can be used in cytotherapy for degenerative diseases of the nervous system and also spinal cord injury

Protective effect of perserved solutions of Krebs contains verapamil, adrenaline and propranololol in comparision with Krebs and Heparinated blood on desqumation of the endothelial cell in Saphenous vein of Guinea pig

Background and Objective: The most important factor in the integrity of saphenous vein is the health degree of endothelium which guaranties the dilation of them after cronary bypass sergery. Kind of preservative soulution has a key role in endothelial protection. This study was done to evaluate Protective effect of perserved solutions of krebs contains verapamil, adrenaline and propranololol in comparision with Krebs and Heparinated blood on desqumation of the endothelial cell in Saphenous vein of Guinea pig. Methods: This experimental study was done on 28 male Guinea pigs with 380±40g weight.for separating 3mm of saphenous vein rings and Measuring of rings nitric oxide released in preserving solutions: Krebs (K), Krebs plus propranolol (K+P), adrenaline (K+A) and verapamil (K+V) compaired with heparinized blood at 30, 45, 60 and 90 minutes after harvesting measured by micro plate Griess reaction. Rings also stained by H&E and examined by light microscopy to evaluate endothelial desqumation. Results: Average concentration of nitric oxide (NO) in the Krebs plus Verapamil solution (K+V) Vs Heparinized Blood (HB), Krebs (K), Krebs plus Adrenaline (K+A) and Krebs plus Propanololol (K+P) revealed significant increase in NO release (P<0.05). The maximum NO measurement was 45 minuts after harvesting. Also histological study with H&E staining showed that endothelial layer was intact only in Krebs plus verapamil in compaired to control group, but in the other solutions the vascular intimal cells had suffered different degrees. Conclusion: It seems that Krebs solutions containing verapamil has more efficiently to the proper functioning of the saphenous veins endothelium in animal modle

An Efficient Protocol for Embryonic Carcinoma Cells P19 Differentiation to Cardiomyocytes Using Oxytocin as Inducer

Background: The capability of embryonic carcinoma cells P19 in differentiation to Cardiomyocyte was examined through inducing effects of Oxytocin (OT) and 5-Azacytidin (5Az) individually and compared with each other in laboratory condition. Materials and Methods: P19 Embryoid Bodies (EBs) was formed through hanging drops method. Then, EBs were treated with (5Az) or (OT) and the EB medium (Ctrl) until 12 days. Morphology and beating number per minute were recorded every two days. Viability was carried out every three days. The expression of several cardiomyocyte-associated genes was assessed by RT-PCR. Results: The beating area percentage of EBs in OT treatment groups was more than that of the 5Az group in all days of experiment. However, only in final stage, a significant increase was observed in beating area of OT group. There was no significant difference in viability and morphological changes. OT induction expressed three more specific proteins in cell culture than 5Az. Conclusion: Statistical analysis revealed that response to OT inducer was more excessive than 5Az in all treatment groups. The Oxytocin was found to be effective inducer of cardiomyocytes differentiation from embryonic carcinoma cells P19 than 5-azacytidine

Common Variations in Prothrombotic Genes and Susceptibility to Ischemic Stroke in Young Patients: A Case-Control Study in Southeast Iran

Background and Objective: Evidence indicates that genetic factors may be involved in the risk of ischemic stroke (IS). The aim of this study was to assess the effect of genetic polymorphisms located in exons or untranslated regions of MTHFR as well as FV genes on ischemic stroke. Materials and Methods: In this case-control study, 106 patients with IS and 157 healthy volunteers (age &lt;50 years) were genotyped for MTHFR C677T, A1298C, C2572A and C4869G, FVL, and prothrombin G20210A polymorphisms. Results: The MTHFR 677CT genotype was more frequent in patients and increased risk of IS with Odds Ratio = 1.9. The MTHFR A1298C and C2572A polymorphisms were not associated with IS in dominant and recessive models. Our findings showed a significant decrease in the MTHFR 4869CG genotype in IS patients, and this variant was associated with a decreased risk of IS in the dominant model. The CAAT haplotype was associated with increased risk, and the GAAC haplotype was associated with decreased risk of IS compared to other haplotypes. There was no relation between FVL G1691A polymorphism and IS risk. Conclusions: The present study showed that the MTHFR 677CT genotype was more frequent and the MTHFR 4869CG genotype was less frequent in young IS patients