JOURNAL OF CELL & TISSUE
Year:2014 | Volume:4 | Issue:4|Papers Count:12

Aim: The aim of this research was to evaluate viability as well as morphological and biochemical featears of apoptosis in human sperm treated with sodium arseinte.Material and methods: Human sperm samples were exposed with different concentration of sodium arsenite (0, 0.1, 20 and 100 mM) for 0, 60, 120 and 180 minutes. Sperm viability was done using MTT assay and sperm DNA integrity was investigated by acridine orange staining. Morphological feature of apoptosis in sperm nucleus was studied using Hoechst and Diff-Quick staining, whereas biochemical feature of apoptosis was studied by TUNEL method. Statistical analysis was carried out by one-way ANOVA and repeated measure.Results: MTT assay showed a significant interaction between sodium arsenite concentration and time on the decrease sperm viability. In addition, after 180 minutes, the viability of sperm treated with sodium arsenite was significantly decreased compared to control. However, this pollutant had no effect on DNA integrity as well as morphological and biochemical features of apoptosis in the sperm nucleus.Conclusion: Sodium arsenite caused a significant decrease in human sperm viability and this effect was not due to the damage of sperm nucleus and DNA.

Aim: Cisplatin is one of the most successful drugs to fight many cancers, but its nephrotoxicity limits its administration. The aim of this study is investigation of the effect of human mesenchymal stem cells secretory factors on Cisplatin-induced acute renal failure in rats.Material and Methods: In this study, male wistar rats were used. Animals were randomly divided into four groups and six animals were in each group. One group was considered as control which this group received no injections. Other three groups were treated with intraperitoneal 5 mg/kg Cisplatin once at the beginning of the experiment. Then one of these three groups was injected with conditioned medium of human mesenchymal stem cells and other groups were injected with medium without secretory factors intraperitoneally for three consecutive days. After five days Cisplatin injection, kidney and blood samples from all of examined animals were collected for histological and biochemical studies.Result: Results showed significant decrease in the kidney injury level, blood serum BUN and creatinine amount. Also Also body weight increasing significantly was observed in conditioned medium of human mesenchymal stem cells receiver group in comparison with treated Cisplatin and non secretory factors receiver groups.Conclusion: Results of this research suggest that human mesenchymal stem cells secretory factors can be affect against Cisplatin-induced nephrotoxicity.

Aim: The aims of this study were subcloning of Dj-1 (PARK7) gene to lentiviral transfer vector, recombinant lentiviruses production and target cells infection with these viruses.Material and methods: DJ1 gene obtained from pcDNA-DJ1 vector using restriction enzymes EcoR1 and Xho1. Lentiviral transfer vector was coincidental digested using EcoR1 and Sall enzymes. DJ-1 gene was interred into the lentiviral transfer and upstream of Jred gene using T4-DNA ligase. As DJ1-IRES-Jred sequence was placed in downstream and control of CMV promoter. To production of recombinant lentivruses, the made vector is coincidental transferred into the HEK-293T (Human Embryonic Kidney) cells with two packaging and envelope lentiviral vectors. Produced virus was used for target cells infection.Results: For integrity of subcloning, enzymatic tests and PCR were used.Fluorescent microscopy also used to show expression of Jred reporter gene that showed the success of our gene transferring. Then RT-PCR was done for showing overexpression of DJ1 gene in transduced cells compare with normal cells.Conclusion: This study show the lentiviral vectors success in gene transferring to eukaryotic cells and clear that this vectors can be used in treatment of nervous system diseases.

Aim: Vitamin D has an important role in treatment and prevention of metabolic syndrome which may enhance the benefits of sportive training. The purpose of this study was to determine the effects of 8 weeks of aerobic training with and without vitamin D supplementation on metabolic syndrome parameters in sedentary women.Material and Methods: In this semi-experimental study with pretest–post designs, thirty obese middle-age women (aged 44.25±5.4 y) were randomly assigned to aerobic training only (RG), vitamin D supplementation (50000IU/w) plus aerobic training (VR) and control (non-exercising=CG) groups. Aerobic training program was performed 50-60 min/d, 3d/wk, for 2 months. Serum levels of 25-hydroxyvitamin D, body composition and metabolic parameters were assessed before and after the training period.Results: Vitamin D supplementation increased serum 25-hydroxyvitamin D levels significantly (P<0.05). Aerobic training caused significant improvement in body weight, waist circumference, HDL-cholesterol, blood glucose and insulin resistance index (P<0.05). Vitamin D supplementation in conjunction with aerobic training lead to greater improvement in adiposity indices, HDL-cholesterol, LDL-cholesterol, total cholesterol, triglyceride, blood glucose and insulin resistance index (P<0.05).Conclusion: These results suggest that aerobic training is a safe and effective for improving cardiometabolic health in middle-age women. Also vitamin D combination can enhance some of the beneficial effects of training over a two-month period.

Aim: Since applied and molecular studies are increasing in Ranunculaceae, the study and understanding of the flower development in the family is necessary. In this research ovule developmental stages in Adonis flammea were studied.Material and Methods: The young flowers and buds were sampled in different developmental stages, fixed in FAA70 and kept in 70% ethanol. Samples were embedded in paraffin after dehydration and sliced using microtome. Staining was carried out with PAS and Hematoxylin. Prepared slides of different developmental stages were studied using light microscope and are photographed.Results: Results showed that ova in A. flammea are anatropous, bitegumic and crassinucellate. Micropylar channel was constructed with inner integument. Megasporocyte produces both linear and nonlinear arrangement of tetrads after meiosis. Embryo sac development follows to the Polygonum type. Antipodal cells are persistent that degenerate at the cellular endosperm stage. Cytoplasmic connections of embryonic sac cells, degeneration of some ova, consciously abortion of ovules and embryo were also observed.Conclusion: The patterns of ovule and embryo sac development are accordance with the Angiosperms and Polygonum type. But antipodal cells persistence, nucellar cap formation, cytoplasmic connections between embryo sac cells, fertilization failure and embryotocia in some ova are characters of the studied species that can due to genetical or environmental factors.

Aim: In this study, the teratogenic effects of Gentamicin on newborn spermatic cords of rabbits whose mothers received this medication during pregnancy is investigated.Material and Methods: The experimental study was done on forty rabbits 6-month old that were divided into 4 groups of 10 (8 females and 2 males) and polygamously impregnated.1.7/mg/kg dosage Gentamicin injections were done in gluteal muscles of pregnanted rabbits in the following groups: 1. control (did not receive any medication), 2. predifferentiation (from 1th to 8th pregnancy days), 3. embryogenic period (from 9th to 16th) and 4. fetal period (from 20th to 27th) for 8 days. After birth, three days old male newborns testes were dissected. Samples were stained using H& E method and light microscopy studies were done.Results: Gentamicin injection to pregnant mother before embryonic differentiation was caused canalization of spermatic cords and their conversion to seminiferous tubules in newborn. Also embryonic stage injection increased seminiferous tubules diameter, but was not observed any changes in newborn whose mother had received medication in embryonic Predifferentiation.Conclusion: Results showed that using Gentamicin for pregnancy infection treatment especially before differentiation and embryonic stages were effective on rabbit newborn spermatic cords morphogenesis and caused early maturation.

Aim: The main goal was to evaluate the impact of different incubation time intervals on human sperm DNA status using terminal deoxyribonucleotidyl transferase–mediated dUTP nick-end labeling (TUNEL) test.Material and methods: This prospective study involved 21 normozoospermic specimens. After direct swim-up, sperm cells were incubated at 37oC and DNA damage was evaluated at different time intervals (0, 1, 2 and 3 h). After slide fixation with methanol 100% for 4 minutes and rinse in phosphate-buffered solution (PBS), TUNEL was added to each slide and incubated for 60 minute in dark. Eosin-Nigrosin and Papanicolaou staining protocols were applied in order to assess sperm viability and morphology, respectively.Results: Sperm viability and normal morphology was improved after sperm processing (100%, 72.3% respectively, p<0.0001). The rate of DNA damage was significantly higher after 2h compared to 0h (9.19±0.8% Vs 4.9±0.9%, respectively, p=0.008). Also there was significant difference in abnormal sperm DNA between 3h and 1h (10.95±0.7% Vs 7.1±1%, respectively, p=0.020).Conclusion: Incubation of prepared normozoospermic samples at 37oC more than 2 h may be associated with sperm DNA fragmentation. Therefore, it seems that incubation of human spermatozoa at 37oC should be limited up to 2h prior to use in ART clinics.

Aim: The aim of this project is finding a relationship between body characters of Hirudo orientalis with testis sac and spermatic follicles structure in order to find a relationship between these characters and sexual maturity, regarding farming goals.Material and methods: 30 individuals of H. orientalis were collected From 3 ponds located in Gonbad-é-Kavous city in May, July and November. Body length, width and weight, Testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers obtained for each sample. For understanding starvation effect on testis sac, 20 individuals of collected leeches were starved for 120 days. Data were analyzed using ANOVA test and SPSS 16 software.Results: Body length, width and weight, testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers showed significantly reduction in May comparing with July and November (P<0.05). Mature spermatic follicles were observed in samples with more than 3 gr weight, >10.3 cm length and>0.79 cm diameter in July and November. All of studied characters were reduced with the exception of body length and testis sac diameter to body diameter ratio in starved samples (P<0.05).Conclusion: This study showed that testis sac structure and spermatic follicle numbers can directly related to body weight and diameter in H. orientalis but not to body length.

Aim: Curcumin, a yellow pigment obtained from turmeric (Curcumina longa). It is a dietary polyphenol that has been reported to possess anticancer, anti-inflammatory, and antioxidant properties. Curcumin is potent radical scavenger and has key role in cell death prevention and because of its antioxidant ability, curcumin has possible role in protection against neurodegenerative diseases. In this study we evaluate the effects of curcumin on rat embryonic neuroprogenitor cells viability and proliferation.Material and methods: In this study brain cortex of 15.5 day old embryos of Wistar rat dissected out and enzymaticaly dissociated to form single cell suspension. Cell suspension seeded in DMEM/F12 medium supplemented with N2, 1% antibiotic and mitogenes (10 ng.ml-1 EGF and 20 ng.ml-1 b-FGF). Cells were treated with different concentrations of curcumin (0.1, 0.5, and 1mM). Cell viability was evaluated with MTT assay and immunocytochemical analysis was performed to evaluate astrocytic marker (GFAP). The data were analyzed with One-way ANOVA with Tukey’s post hoc test.Results: curcumin (0.5 mM) enhanced astrocytes neurite size (p<0.001) and also has significante effects on neuroprogenitor cells differentiation in culture media.Conclusion: Data suggest that curcumin enhanced neuroprogenitor cells viability, growth and promoted differentiation of these cells in to astrocytes.

Aim: The aim of study was designed to determine the created macroscopic abnormalities by Gabapentin during of implantation and organogenesis of fetus.Material and methods: In this experimental study forty female mice (NMRI type) were used. Female pregnant mice were divided in four groups (each one 10) consist of three experimental groups (I, II, III) and one control (received normal saline). Three experimental groups received 15, 30 and 45 mg/kg/day of Gabapentin respectively by intraperitoneally (ip) injection from day 6.5 (GD6.5) to day 14.5 (GD14.5) of pregnancy. Animals were dissected in 18.5 day of pregnancy and their embryos were studied using stereomicroscope from the macroscopical abnormalities. Embryos weight and length were measured and recoded based on CR length. Data were analyzed by T-test, Tukey test and ANOVA using SPSS software.Results: In all three experimental groups (I, II, III) was observed a significant reduction in comparison with control in body weight and CR length (P<0.001). Also significant increasing was observed in abnormalities appearance such as fetal absorption, organs bleeding and follicular thyroid in comparison with control.Conclusion: Gabapentin drug is caused fetal absorption, weight and CR length reduction in mice embryo during organogenesis. Also it induces some abnormalities such as hemorrhage and follicular thyroid in embryonic period.