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Assessment of Motility and Survival Rate of Asthenospermic Men's Sperm Cultured in Media Plus Leukemia Inhibitor Factor
Fakher Rahim,Ghasem Saki
International Journal of Pharmacology , 2010,
Abstract: As our knowledge no report was given about effect of leukemia inhibitor factor on sperm motility and survival rate of asthenospermic infertile men. That's why this study was decided to review the effects of Leukemia Inhibitor Factor (LIF) with different concentrations of 0, 3, 5, 10, 50 ng mL-1 on motility and survival rate of asthenospermic infertile men. Semen samples of 15 asthenospermic men who referred to IVF unit of Imam Khomeini Hospital, Ahvaz, Iran were collected and put in incubator under condition 5% CO2 in air at 37°C for 45-30 min then total sperm count was done first, then calculate the motile sperm with a degree a and b. In this study, we evaluated only samples that had sperm motility percent of more than 30%. After that time from every sample about 10 μL removed and culture in different media. Every drop was evaluated 6, 24, 48 h after cultured of sperm in it for motility and survival rate of sperm. Statistical analysis shows that the forward motility of sperm cultured for 6 h in media with and without of LIF is not significant (p>0.05) but after 24 h the forward motility and survival rate of sperm cultured in media with 10 and 50 ng mL-1 of LIF significantly increased (p<0.05) and after 48 h the forward motility and as well as survival rate of sperm cultured in media with 50 ng mL-1 of LIF significantly increased (p<0.05). The conclusion from this study is that certain concentrations of leukemia inhibitor factor can increase the motility and survival rate of sperm.
Endocrine Function and Duration Time of Estrous Cyclicity of the Ovariectomized Recipiented Neonate Vitrified Ovarian Grafts Mice after Treatment with Melatonin
Hemadi Masoud,Saki Ghasem
International Journal of Pharmacology , 2010,
Abstract: e melatonin administration reduced these high levels into nearly similar concentrations to those in intact mice. The correlation coefficients between gonadothropins and melatonin concentrations at the different stages of the estrous cycle were significantly different from zero. indeed, progesterone secretion in spite of estradiol was adversely affected by melatonin treatment. Meanwhile, the correlation coefficients were significantly different from zero. These results suggest that melatonin could be having positive effects on the deficient activity of hypothalamic-pituitary-ovarian axis especially on the progesterone drive of the recipient.
The Study of Developmental Capacity of Vitrified Mouse Blastocysts in Different Straws after Transfer to Mouse Pseudo Pregnant
Ghasem Saki,Fakher Rahim,lida Moradi
Pakistan Journal of Biological Sciences , 2008,
Abstract: Vitrification is the commonly used method for long-term storage of pre-implantation mammalian embryos. It is an essential part of assisted reproductive technologies. The re-expansion rate, pregnancy and birth rate of vitrified blastocysts using CPS were compared with OPS and Conventional Straw. Female NMRI mice were injected with Gonadotrophins in order induce them for super ovulation. At that time the mice were sacrified by cervical dislocation and dissected of mouse abdomen. The uterine horns were existed blastocysts were collected in PBS and randomly allocated to four groups: vitrification in CPS, conventional straw, OPS and untreated controls. The vitrification solution was EFS40%. After storage for 1 month in liquid nitrogen, the blastocysts were thawed in 0.5 M sucrose for in vitro culture in M16 medium. After 6 h of culture, the numbers of expanded blastocysts was recorded and ready for transfer to uterus of pseudo pregnant mouse. The re-expansion rate of the CPS group (72.1%) was significantly higher (p<0.05) than OPS (52.55) and C.S. (38.6%) groups. The pregnancy (70%) and birth rate (45%) of blastocysts in CPS were similar to those of fresh blastocysts (80% and 45.5%) and the pregnancy (10%) and birth rate (5.1%) in Conventional Straws lower than OPS (20 and 7.5%), but were not significantly different. Mouse blastocysts vitrified using CPS had a better result compared with OPS and Conventional Straw. The value of CPS for vitrification of blastocysts may also merit investigation.
Effect of forced swimming stress on count, motility and fertilization capacity of the sperm in adult rats
Saki Ghasem,Rahim Fakher,Alizadeh Karim
Journal of Human Reproductive Sciences , 2009,
Abstract: Aims: The purpose of this study was to determine whether 50 days of forced swimming stress applied to adult male rats affects count, motility and fertilization capacity of sperm. Settings and Design: It is a prospective study designed in vitro. Materials and Methods: A total 30 adult male wistar rats were used in this study. All rats were divided into two equal groups (n = 15): (1) control group and (2) experimental group. Animals of the experimental group were submitted to force swimming stress for 3 min in water at 32°C daily for 50 days. Then, all male rats were sacrificed, the right epididymides were removed and sperm concentration and motility were determined. The sperm suspension was added to the ova. Fertilization capacity was assessed by counting two-cell embryos 24-26 h after completion of fertilization in vitro. Statistical Analysis Used: Data are reported as mean ± SD and percentage. The difference between the control and experimental groups was determined by the unpaired t-test. Results: The mean and standard deviation of sperm concentration in the control and experimental groups were 60.8 ± 9.3 10 6 /ml and 20.4 ± 5.3 10 6 /ml, respectively. There was a statistical difference of P < 0.05 between the two groups in terms of sperm concentration. The percentage of motility in the experimental group was significantly different ( P < 0.05). The same results were obtained in case of fertility ( P < 0.05). Stress caused by forced swimming was observed by a significant increase in the latency of the pain response in the hot-plate test ( P < 0.05). Conclusions: These results suggest that forced swimming stress in time course equal or more than spermatogenesis period, i.e. 48-50 days in the rat will be significantly effective to reduce the number and motility of sperms as well as the fertilization capacity.
Effect of forced swimming stress on in-vivo fertilization capacity of rat and subsequent offspring quality
Saki Ghasem,Rahim Fakher,Vaysi Ozra
Journal of Human Reproductive Sciences , 2010,
Abstract: Aims: This study aimed to determine the effect of 50 days of forced swimming stress on fertilization capacity of rat and subsequent offspring quality. Setting and Design: The prospective study designed in vivo. Materials and Methods: Total 90 Wistar rats including 30 adult male (3 months of age, weighing 210 ± 10.6 g) and 60 female rats (3 months of age, weighing 230 ± 12.2 g) were engaged in this study. Male rats were randomly divided in two equal groups (n=15): Control and experimental groups. Animals of the experimental group were submitted to forced swimming stress for 3 min in water at 32oC daily for 50 days. Then all adult male rats were mated with normal females (2 per each male) for 7 days. Female rats were sacrificed and autopsy was performed on day 20 of pregnancy when uterus and ovaries were examined for the number of corpora lutea, dead and live fetuses, embryo resorption, implantation sites, and fetus weight. Conclusion: Results of this study have important implications for families attempting pregnancy. Stress pursuant to life events may have a negative impact on in vivo fertilization capacity of male rats and subsequent offspring quality.
Vitrification of Small Volume of Normal Human Sperms: Use of Open Pulled Straw Carrier
Ghasem Saki,Fakher Rahim,Majied Jasemi Zergani
Journal of Medical Sciences , 2009,
Abstract: The objective of this study was to evaluate whether cryopreservation of small volume of sample (sperm+cryoprotectant) was feasible using open pulled straw and also compared the outcomes of open pulled straw and conventional straw as carrier for normal human sperm cryopreservation. Semen samples were obtained from 10 men undergoing evaluation for infertility after 3-4 days of abstinence. Washed normal sperm samples were divided into three aliquots as follows: (1) Fresh; (2) cryopreserved in open pulled straw and (3) crypreserved in conventional straw. In order to do cryopreservation of sperm in open pulled straw first washed normal sperm samples were mixed with equal volume of test yolk buffer and 12% v/v glycerol, later on 3-4 μL from the prepared mixture was loaded in each straw by using syringe. The loaded straws were plunged into liquid nitrogen and after 3 months recovered and thawed. Each straw was emptied of their fluid content in drop of 10 μL medium covered with mineral oil. Motility of vitrified-thawing sperm was assessed by using inverted microscope. The results show as percent progress motility±SD and the p<0.05 were suggested as significant. The percent progress motility±SD of fresh sperm was evaluated as follow in study groups: For fresh group was evaluated as 59.2±7.6; the value of 37.5±8.2 in cryopreserved sperm in open pulled straws group; and value of 26.3±6.4 for conventional straw group, respectively. Statistical analysis shows that the difference between cryopreserved sperm in open pulled straws and conventional straw groups is significant (p = 0.001). Because of the significant difference between cryopreserved sperm in open pulled straws and conventional straw groups, we concluded that vitrification of human sperm is feasible using open pulled straw. The results of this study shows that open pulled straw could be a good carrier for cryopreservation of small volume of normal human sperm.
Evaluation of Nanohydroxyapatite Powder Synthesized by Sol-gel Combustion Method and Bio Oss on Reconstruction of Parietal Bone Defects in Rat
Iraj Kazeminezhad,Mahmood Jahangirnezhad,Ghasem Saki,Maryam Rahimzadeh Larki
Jundishapur Scientific Medical Journal , 2013,
Abstract: Background and Objective: The aim of this research is to investigate the possible use of hydroxyapatite nanoparticles in parietal bone regeneration. Subjects and Methods: Ca10(PO4)6(OH)2 nanoparticles were synthesized by sol-gel combustion method using citric acid as combustion agent. The calcium nitrate tetrahydrate, citric acid and ammonium dihydrogen phosphate and deionized water with the proper stoichiometry were used. The nanoparticles were studied by the TG/DTA, XRD, FT-IR, SEM and TEM techniques. XRD. Fourier transform infrared spectroscopy (FT-IR) was used to identify different bond groups in the structure of the samples. The hydroxyapatite nanopowders were used on of adult male Sprague Dawley rats for parietal bone regeneration. Results: The product had a hexagonal structure. SEM images showed morphology and mean nanoparticles size. Conclusion: The results showed that the bone repairing with HA nanoparticles occur similar to typical repairing by commercially available (Bio Oss) and even it takes place with minimal tissue inflammation.
Study of Spermatogenesis Fetal Testis Exposed Noise Stress During and after Natal Period in Rat
Maryamalsadat Jalali,Masoud Hemadi,Ghasem Saki,Alireza Sarkaki
Pakistan Journal of Biological Sciences , 2013,
Abstract: Noise stress is dangerous natural contaminant that produces harmful physiological, psychological and morphological outcomes to the body. So this study was conducted in order to investigate the effects of noise stress on the parenchyma of testis. Healthy mature females rats (n = 20) were mated with the mature male rats and then randomly allocated equally either to experimental or control groups. Experimental group has given daily noise stress up to birth their child. In the second step, the child's pregnant rats of experimental group were distributed to three subgroups as follow: group I (without exposure to noise stress), group II (exposure to noise for 8 weeks) and group III (exposure to noise for 14 weeks) for morphometric analysis of their child's testicles by sacrificing of them at weeks 14. In general, the testes of non-exposed group were grown larger than ones in the noise exposed groups. Moreover, the testes of the experimental group 1 were larger than the other experimental groups. Indeed, the rate of atrophic seminiferous tubules and jumbled appearance of the interstitial space were more observed in the noise stress exposed group than non-exposed ones. In addition, seminiferous tubules analysis revealed that the characteristics of interstitial space cells and epithelial germinative cells of the seminiferous tubules in the control group were better than the noise exposed groups. It seems that the noise stress has negative influences on the fertility of male based on enhancing of the apoptotic process induced by pathogenesis stress and suppressing the kinetics spermatogenesis.
Ultrastructural Change of Cerebellum in Exposed Rats to 3mT Electromagnetic Field
Allahvaysi Ozra,Solaeymani-Rad Jafar,Lida Moradi,Ghasem Saki
Journal of Biological Sciences , 2010,
Abstract: The aim of this study was to investigate ultrastructural changes of Cerebellum in 3mT electromagnetic field exposed rats. Total 30 adult female Wister rats with 3 months of age and weighing 210±10.6 g were used in this study. All female rats subdivided randomly to 2 groups: group 1, serve as untreated controls; group 2, was exposed to 3mT EMF for 4 months, 4 h day-1. After 120 days all rats were killed and their tissue samples from Cerebellum were removed and prepared for electron microscopic studies. Present finding clearly demonstrated that number of purkinje cells in the cerebellum of EMF- exposed rats were decreased significantly (p<0.01) in comparison to control group. The other changes include: condensation of nuclei, dilatation of endoplasmic reticulum, breakdown and disappearance of crista in mitochondria and vacuolization of cytoplasm in the purkinje cells of cerebellum. The mean nuclear diameter in purkinje cells were 45.35±22.85 mm and 26.79±16.36 mm in control and experimental group respectively. The statistical analysis showed that the difference between two group was significant (p = 0.03). Axial ratio of nucleus of purkinje in control and experimental groups were 1.86±0.41 and 1.55±0.14 mm, respectively. The axial ratio of nucleus in purkinje of EMF-exposed cerebellum were decreased significantly in comparison to control group (p = 0.02). These findings indicate that long-term exposure to EMF has detrimental effects on central nervous system at cellular level.
DARU : Journal of Pharmaceutical Sciences , 2004,
Abstract: In this study effect of recombinant human leukemia inhibitory factor on invitro development of 2 cells embryos and isolated blastomeres derived from mouse 2 cell embryos were investigated. Female ICR mice that were between 8 to 10 weeks old received intraperitoneal injection of 7.5 IU of PMSG for super ovulation followed by intraperitoneal administration of 7.5 IU of HCG 48 hours later. The mice were then mated to mature ICR male mice and were checked for vaginal plugs after 13-14 hours. Mice were killed 46-48 hours after HCG injection by cervical dislocation, their oviducts were removed and flushing 2 cell embryos were collected. The zona pellucida of 2 cell embryos were removed by Acid Tyrod solution and blastomeres separated with oocyte preparation pipette and then all embryos and blastomeres were cultured in Potassium Simplex Optimized Medium (KSOM) +Aminoacid (AA) different amounts of rhLIF (500IU/ml, 1000IU/ml and 1500IU/ml). Some embryos and individual blastomere also were cultured without rhLIF as control group. All samples were cultured in an incubator at 370C with 0.05 CO2 for 120 hours. The rate of embryo and individual blastomeres which reached to 2 cell, 4 cell, 8 cell and 9-16 cell were the same in all groups. However in further developmental stages, morula and blastocyst between experimental and control groups were significantly different. Therefore it may be concluded that: cultivation of isolated blastomers up to the blastocyst stage with rhLIF has stimulatory effect on the preimplantation stage (morula and blastocyst) but it has no stimulatory and inhibitory effects when was added to culture media at the early cleavage stage.
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