فهرست مطالب aligholi sobhani

The current research was established to make a comparison between the delayed-start GnRH antagonist and flare-up GnRH agonist protocols in poor response patients.

The present study is a randomized, prospective, controlled trial that was performed on 150 women who referred to two distinct in vitro fertilization (IVF) centers in Iran. Patients were randomly assigned to two experimental groups, as one group was treated with the delayed-start GnRH antagonist protocol (delayed-start group), while another group was treated with the flare-up protocol (flare-up group).

The serum concentrations of estradiol and progesterone, along with the thickness of endometrial tissue and the number of follicles ≥13 mm was significantly increased in the delayed-start group compared with the flare-up group. Also, the number of total oocytes, retrieved mature oocytes, total embryos, fertilized oocytes, as well as the quality of embryos were markedly higher in the delayed-start group when compared with the flare-up group. No statistically significant difference was found in the rates of fertilization, implantation, and pregnancy between the two experimental groups.

According to the above evidence, it seems that the effect of delayed-start protocol on ovarian responsiveness was more pronounced during controlled ovarian stimulation in comparison with the flare-up protocol and the delayed start protocol probably lead to better implantation and pregnancy rates in comparison with the flare up agonist protocol cycle in poor responders.

The cases with unexplained infertility may have an abnormality in their sperm chromatin structure. Sperm selection methods can be used to separate sperm with low DNA fragmentation. The purpose of this study was to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) with magnetic- activated cell sorting (MACS) in assisted reproductive techniques in cases with un- explained infertility.

The semen samples were collected from couples with unexplained infer- tility. After semen analysis and sperm DNA fragmentation (SDF) evaluations, sam- ples were prepared with swim-up method. The rates of SDF in different fractions in- cluding raw semen (n=20), swim-up (n=20), only motile sperm after swim-up (swim-up selection) (n=20), MACS sperm selection (n=20), only motile sperm after MACS (MACS selection) (n=20), and PICSI sperm selection (n=16) were evaluated. Also, the main sperm characteristics and fine morphology of sperm suspension after MACS were assessed. Statistical analysis was performed using GraphPad Prism. The p<0.05 was considered statistically significant.

DNA fragmentation index (DFI) values in PICSI and MACS groups were significantly reduced as compared to the swim-up group. The rate of this reduction was more pronounced in MACS (58.20±13.02) than PICSI (36.57±15.52) group. Al- so, our results showed that MACS resulted in decreased sperm motility, with no al- teration in their fine morphology.

MACS was found to be more efficient in reduction of SDF rates than PICSI. However, none of the sperm selection techniques can not totally eliminated the spermatozoa with DNA fragmentation in the final sperm sample.

In vitro development of functional gametes from pluripotent stem cells is a promising prospect to treat infertility. Mesenchymal stem cells with a high degree of plasticity and less tumorigenicity are a reliable source of stem cells for the generation of gametes. The present study aimed to compare the differentiation potential in the mesenchymal stem cells that are derived from bone marrow (BMDMSCs) and adipose tissue-derived mesenchymal stem cells (AD-MSCs) into germ cells in a culture medium containing bone morphogenic protein-4 (BMP-4).

Experimental approach: 

In this study, MSCs were isolated from both bone marrow and adipose tissue of murine samples. To further verify the nature of the harvested stem cells, their multipotency and surface marker were examined. The identified stem cells were cultured in a medium supplemented with 0 and 25 ng/mL of BMP-4 for 4 days. Flow cytometry analysis, immunofluorescence staining, and real RT-PCR were used to assess the expression levels in germ cell-specific biomarkers (Mvh, Dazl, Stra8, and Scp3).

CD44+, CD45−, CD31−, BMD-MSCs, and AD-MSCs showed to be capable of differentiating to osteo-adipogenic lineages. The flow cytometry, immunofluorescence, and RT-PCR results indicated that early germ cell markers (Mvh and Dazl) were expressed in both types of cells but they were significantly higher in BMD-MSCs than AD-MSCs.

Conclusion and implications: 

Based on our results, the addition of exogenous BMP4 to the culture medium could differentiate BMD-MSCs and AD-MSCs into primordial germ cells, but it is inadequate to further develop into late germ cells in vitro. Moreover, the results revealed that, although AD-MSCs were easier to collect and had faster growth and proliferation rates than BMD-MSCs, the BMD-MSCs were better capable of differentiation into primordial germ cells. They may serve to be considered a more suitable source of MSC for in vitro generation of gametes than AD-MSCs.

Astaxanthin (AST) has been introduced as a radical scavenger and an anti-apoptotic factor that acts via regulating the nuclear factor-E2-related factor 2 (NRF2) and related factors. Here, we intended to examine the effect of AST on granulosa cells (GCs) against oxidative stress by examining NRF2 and downstream phase II antioxidant enzymes.

In this experimental study, we used cultured human primary GCs for the study. First, we performed the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test to evaluate cells viability after treatment with hydrogen peroxide (H2 O2 ) and AST. The apoptosis rate and ROS levels were measured by flow cytometry. To determine NRF2 and phase II enzymes expression, we performed real-time polymerase chain reaction (PCR). Finally, we used western blot to measure the protein levels of NRF2 and Kelch-like ECsH-associated protein 1 (KEAP1). Enzyme activity analysis was also performed to detect NRF2 activity.

This study showed that AST suppressed ROS generation (P<0.01) and cell death (P<0.05) in GCs induced by oxidative stress. AST also elevated gene and protein expression and nuclear localization of NRF2 and had an inhibitory effect on the protein levels of KEAP1 (P<0.05). Furthermore, when we used trigonelline (Trig) as a known inhibitor of NRF2, it attenuated the protective effects of AST by decreasing NRF2 activity and gene expression of phase II enzymes (P<0.05).

Our results presented the protective role of AST against oxidative stress in GCs which was mediated through up-regulating the phase II enzymes as a result of NRF2 activation. Our study may help in improving in vitro fertilization (IVF) outcomes and treatment of infertility.

Rutin (quercetin-3-rhamnosyl-glucoside), a flavonoid, is derived from plants and has antioxidant properties. This study aimed to evaluate the effect of different concentrations of rutin on mouse ovary heterotopic allotransplantation. The present animal experimental study was conducted on 40 female adult Balb/c mice weighing 30 ± 5 g at the Jundishapur University of Medical Sciences, Ahvaz, Iran, during 2016 - 2018. The mice were divided by permuted block randomization into 8 groups (n = 5): OVX (ovariectomy), as the negative control; normal (positive control); OVX + OVA (ovariectomy and transplantation) (control), treated with 0.5 mL of normal saline; OVX + OVA + 10 mg/kg of rutin; OVX + OVA + 30 mg/kg of rutin; OVX + OVA + 60 mg/kg of rutin; OVX + OVA + 100 mg/kg of rutin; and the autograft. Groups were treated daily. Fourteen days after transplantation, ovarian grafts were collected and processed histologically for follicle number counting. Serum estrogen (E2) and progesterone (P4) levels were evaluated. Furthermore, the expression of Estrogen Receptor alpha (ERα), Estrogen Receptor beta (ERβ), and Progesterone Receptor (PR) in the uterine endometrial tissue was tested using qRT-PCR and western blotting. A decrease in the number of mature follicles and increase in the number of atretic follicles (mean ± SD: OVX + OVA + 30 = 19.00 ± 1.000, OVX + OVA + 60 = 25.00 ± 5.000, and OVX + OVA + 100 = 23.00 ± 2.646) were observed in all groups treated with rutin in comparison with the control group (mean ± SD: 12.33 ± 2.517) (P value < 0.05). The level of E2 and P4 (mean ± SD: OVX + OVA + 100 = 6.133 ± 1.026) increased in comparison with the OVX + OVA group (mean ± SD: 0.4667 ± 0.2517) (P value < 0.05). The protein expression of ERα (mean ± SD: OVX + OVA + 10 = 1.615 ± 0.1701 and OVX + OVA + 30 = 1.744 ± 0.1779) in comparison with the control group (mean ± SD: 0.7089 ± 0.1131), and ERβ (mean ± SD: OVX + OVA + 10 = 0.7747 ± 0.4365, OVX + OVA + 30 = 0.9220 ± 0.1245, OVX + OVA + 60 = 0.7701 ± 0.2150, and OVX + OVA + 100 = 0.6676 ± 0.1547) increased in a dose-dependent manner in all groups treated with rutin in comparison with the OVX + OVA group (mean ± SD: 0.1534 ± 0.06109) (P value < 0.05). No significant changes in PR were found in groups treated with rutin in comparison with the control group. The results of the present study indicated that rutin increases E2 and P4 levels in ovarian hetero allograft mice. Rutin also upregulated the expression of ERα and ERβ but had no significant effect on PR.

Contrary to a common belief, most mammalian females lose the ability of Germ Cell (GC) renewal and oogenesis during fetal life. Although, it has been claimed that germ line stem cells preserve oogenesis in postnatal mouse ovaries, that postnatal oogenesis keeps producing functional and sufficient GCs in the case of infertility (caused by different reasons) is doubtful. On the other hand, there are many studies showing derivation of primordial GCs and late GCs from Embryonic Stem Cells (ESCs) in vitro. This study aimed to clarify the role of ESC-derived GCs in oogenesis.
Mouse ESCs via Embryoid Body (EB) formation were differentiated into GC lineage by adding Bone Morphogenetic Protein 4 (BMP4) and Retinoic Acid (RA) to the culture medium. Expression of GC markers was characterized by using Reverse Transcription Polymerase Chain Reaction (RT-PCR) and immunohistochemistry. Several 6- to 10-week-old female mice, sterilized using chemical agents, were injected with ESCs-derived GCs thorough their tail veins. To track the transplanted cells, their ovaries were immunohistochemically stained after two months.
Expression of GC specific markers such as mouse vasa homologue (Mvh) and Deleted in Azoospermia-Like (DAZL) indicated that GCs were successfully developed from ESCs. Interestingly, there was no evidence of homing of GCs in the transplanted ovaries after transplantation of ESCs-derived GCs.
Our findings do not suggest any contribution of ESC-derived GCs within the sterilized mice ovaries.

Stem cells are self-renewing and undifferentiated cell types that can be differentiate into functional cells. Stem cells can be classified into two main types based on their source of origin: Embryonic and Adult stem cells. Stem cells also classified based on the range of differentiation potentials into Totipotent, Pluripotent, Multipotent, and Unipotent. Multipotent stem cells have the ability to differentiate into all cell types within one particular lineage. There are plentiful advantages and usages for multipotent stem cells. Multipotent Stem cells act as a significant key in procedure of development, tissue repair, and protection. Multipotent Stem cells have been applying in treatment of different disorders such as spinal cord injury, bone fracture, autoimmune diseases, rheumatoid arthritis, hematopoietic defects, and fertility preservation.

Stage-specific embryonic antigen-1 (SSEA1) is a cell surface carbohydrate that its pattern expression is changed during induction of mouse embryonic stem cell differentiation. In this study, the spatial distribution of SSEA1 on primordial germ cells differentiation and subsequent progression into oocyte-like cells from mouse embryonic stem cells in vitro was evaluated. Embryoid bodies from mouse embryonic stem cells were cultured for two days with 5 ng/mL BMP4. SSEA1 positive and negative cells were separated using the MACS system and cultured separately in a conditioned medium consist of in vitro maturation medium diluted in DMEM [1:1] for 10 days. We assayed viability, colony formation and alkaline phosphatase activity (ALP) of sorted cells. Also, germ cell markers were analyzed by flow cytometry, Immunocytochemistry and RT-PCR. Viability percent SSEA1 positive cells were more than SSEA1 negative cells. SSEA1 positive cells and SSEA1 negative cells formed compact and flat colonies respectively. Unlike the SSEA1 positive population, the SSEA1 negative colonies showed a weak ALP activity. SSEA1 positive cells expressed Oct4, Stella, Mvh, c-kit, Scp3, Desmin, GFAP and Albumin. Interestingly, SSEA1 negative cells expressed Desmin and GFAP. The population of Mvh-positive cells in SSEA1 positive was 17.74%. All specific oocyte mentioned genes were detected in the SSEA1 positive. Also, oocyte specific proteins GDF9 and ZP3 were detected using Immunocytochemistry. Our results suggest that conditioned medium provides a suitable niche to differentiation and progression putative primordial germ cells derived from the SSEA1 positive toward oocyte-like cells.

The existence of female germ-line stem cells (FGSCs) has been the subject of a wide range of recent studies. Successful isolation and culture of FGSCs could facilitate studies on regenerative medicine and infertility treatments in the near future. Our aim in the present study was evaluation of the most commonly used techniques in enrichment of FGSCs and in establishment of the best procedure. In this experimental study, after digesting neonate ovary from C57Bl/6 mice, we performed 2 different isolation experiments: magnetic activated cell sorting (MACS) and pre-plating. MACS was applied using two different antibodies against mouse vasa homolog (MVH) and stage-specific embryonic antigen-1 (SSEA1) markers. After the cells were passaged and proliferated in vitro, colony-forming cells were characterized using reverse transcription-polymerase chain reaction (RT-PCR) (for analysis of expression of Oct4, Nanog, C-kit, Fragilis, Mvh, Dazl, Scp3 and Zp3), alkaline phosphatase (AP) activity test and immunocytochemistry. Data showed that colonies can be seen more frequently in pre-plating technique than that in MACS. Using the SSEA1 antibody with MACS, 1.98 ± 0.49% (Mean ± SDV) positive cells were yield as compared to the total cells sorted. The colonies formed after pre-plating expressed pluripotency and germ stem cell markers (Oct4, Nanog, C-kit, Fragilis, Mvh and Dazl) whereas did not express Zp3 and Scp3 at the mRNA level. Immunocytochemistry in these colonies further confirmed the presence of OCT4 and MVH proteins, and AP activity measured by AP-kit showed positive reaction. We established a simple and an efficient pre-plating technique to culture and to enrich FGSCs from neonatal mouse ovaries.

In Iranian folk medicine Ruta graveolens has been used for female and male contraceptive. There are few studies about the effect of this plant on spermatogenesis.. In this study the effect of long term administration of aqueous extract of RG on spermatogenesis has been investigated.. Animals were allocated into 1) control: which did not receive anything, 2) vehicle which received only normal saline and 3) experiment: which received Ruta extract (300 mg/kg administered by gavage once a day for 100 days). A day after last gavage all the individuals were killed by euthanasia. The right testes and epididymis were extruded. The sperm motility was assessed and classified as progressive, no progressive.. There was a significant decrease in the number of spermatogonia (P < 0.01), primary spermatocyte (P < 0.05) and spermatid (P < 0.05) in experimental group as compared to control and vehicle. As shown in Table 3 the sperm count ± SD in 1 gram of epididymis was 2597.5 ± 172.39 in vehicle, 2671.8 ± 38.57 in control groups and 607.4 ± 520.19 in experimental group. Therefore group 3 has a significant lower sperm count in comparison with other groups (P < 0.05). Sperm with progressive motility was 55.25 ± 2.81 in vehicle, 53.42±1.82 in control group and 11.16 ± 2.17 in experimental group. Statistical analysis show that rats in experimental group have a significant lower sperm motility in comparison with other groups (P < 0.05). There was no difference between other groups (P > 0.05).The fertilization capacity of sperm of rats in experimental group was significantly lower than other groups (P > 0.05).. It is concluded that the aqueous extract of Ruta graveolens diminishes the reproductive system activity and might be a useful substance for birth control process..

Mesenchymal Stem Cells (MSCs) are multipotent cells that can be collected from different sources. Under specific conditions, MSCs can be differentiated to tissue specific cells in vitro. Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) can easily be harvested and cultured in in vitro conditions. Production of germ cells from mesenchymal stem cells is a very interesting and promising area in the field of reproductive medicine. In the present study, the possible trans-differentiation of hUCMSCs into Primordial like Germ Cell (PGC) was performed in vitro under specific condition. Human umbilical cord mesenchymal stem cells were cultured and expanded in DMEM medium containing 10% FBS. The cultured cells were studied for differentiation ability to adipocytes and osteocytes. Furthermore, MSCs related markers were identified by flow cytometry method. For PGC differentiation, hUCMS cells were cultured in differentiation medium containing Bone Morphogenetic Protein 4 (BMP4) and it was followed by retinoic acid (RA). Real time PCR and immunocytochemistry analysis were performed to evaluate the expression of PGC specific genes and proteins, respectively. Our results showed that hUCMSCs cultured in the presence of BMP4 and RA are able to transdifferentiate in to PGC like cells in vitro. Real time PCR and immunocytochemistry results showed that differentiated cells expressed PGC specific markers after 14 days of culture. Based on these results, it was concluded that hUCMSC may be considered as a promising alternative cell source in reproductive medicine. More studies including laboratory and also animal models are needed to evaluate the functionality of differentiated PGCs before introducing them to clinical applications.

Recent publications regarding the differentiation of stem cells to germ cells have motivated researchers to make new approaches in infertility. In vitro production of germ cells improves the understanding of differentiation process of male and female germ cells. Since using embryonic stem cells for this purpose has been associated with tumorogenesis and ethical criticisms، the mentioned cells were suggested to be replaced with some adult multipotent stem cells. In this study، we used Retinoic acid to induce the differentiation of stem cells into germ cells. To find appropriate non invasive source replacement for embryonic stem cells in this study; we evaluated differential potentials of Adipocyte Derived Mesenchymal Stem Cells (ADMSCs) to germ like cells. To find the differentiation capabilitypurified ADMSCs were obtained and differentiated to osteoblasts and adipocytes by using appropriate culture medium (Alizarin Red S and Oil red-O). Superficial markers for mesenchymal stem cells (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flow cytometry to confirm mesenchymal lineage production. The cells were differentiated to germ cells in mediums containing Retinoic acid for 7 days. To evaluate germ cells characteristic markers (Mvh and Dazl) flow cytometry and imunoflorescence were used. Presence of stem cell superficial markers (CD90 and CD44) and absence of endothelial and blood cell markers (CD31 and CD45) were confirmative for the mesenchymal origin of these cells. The cells were able to differentiate into osteoblast and adipocyte cells. This fact was representative for the multipotential entity of the examined cells. After treating the cells with Retinoic acid، flow cytometry and imunoflorescence results showed remarkable expression of germ cells characteristic markers (Mvh and Dazl). By this study، it was found that germ cell markers were expressed in ADMSCs after adding exogenous Retinoic acid into culture medium.

Recent publications about differentiation of stem cells to germ cells have motivated researchers to make new approaches to infertility. In vitro production of germ cells improves understanding differentiation process of male and female germ cells. Due to the problem of using embryonic stem cells (ESC), it’s necessary the mentioned cells be replaced with some adult multi-potent stem cells in laboratories. The aim of this study was to obtain germ cells from appropriate source beyond ESC and compare differential potentials of adipocytes derived stem cells (ADMSCs) with bone marrow derived stem cells (BMMSCs). To find multi-potential entity, after providing purified ADMSCs and BMMSCs, differentiation to osteoblast and adipocyte was confirmed by using appropriate culture medium. To confirm mesenchymal lineage production superficial markers (expression of CD90 and CD44 and non-expression of CD45 and CD31) were investigated by flowcytometry. Then the cells were differentiated to germ cells in inductive medium containing retinoic acid for 7days. To evaluate germ cells characteristic markers [Dazl (Deleted in azoospermia-like), Mvh (Mouse vasa homolog gene), Stra8 (Stimulated by retinoic acid) and Scp3 (Synaptonemal complex protein 3)] flowcytometry, imunoflorescence and real time PCR were used. Both types of cells were able to differentiate into osteoblast and adipocyte cells and presentation of stem cell superficial markers (CD90, CD44) and absence of endothelial and blood cell markers (CD31, CD45) were confirmative The flowcytometry, imunoflorescence and real time PCR results showed remarkable expression of germ cells characteristic markers (Mvh, Dazl, Stra8, and Scp3). It was found that although ADMSCs were attained easier and also cultured and differentiated rapidly, germ cell markers were expressed in BMMSCs significantly more than ADMSCs.

Cryopreservation and in vitro maturation (IVM) of oocyte is becoming an important technique in infertility treatment and fertility preservation. Also it has been proposed to establish a genetic resource bank for endangered or commercially important animal species. The aim of this study was to evaluate viability, maturation and fertilization rate of mouse immature oocytes after single and stepwise vitrification procedure. Oocytes were obtained from 4 weeks old female mice 48h after intraperitoneal injection of 7.5 IU pregnant mare serum gonadotropin (PMSG). Collected oocytes before vitrification were exposed to cryoprotectant, which was composed of 30% (v/v) ethylene glycol, 18% (w/v) Ficoll-70, and 0.3 M sucrose, either by single step or in a step-wise way. After vitrification and storage in liquid nitrogen, the oocytes were warmed and washed two times in medium TCM199 and then subjected to IVM, fertilization and subsequent development to blastocysts. The oocytes survival rates after vitrifying-warming (88.96%), maturation rate (73.23%), the capacity of fertilization (57.80%) and embryonic development to blastocyst (16.41%) in the step-wise exposure were significantly higher (p<0.001) compared with corresponding rate in the single step procedure. The results suggest that vitrification with step-wise procedure has positive effects on maturation and developmental capacity of mice germinal vesicle oocytes in compare with single step vitrification procedure.

Bone morphogenetic protein 4 (BMP4) has a significant role in primordial germ cells (PGCs) differentiation from mouse embryonic stem cell (mESC). The aim of this study is to determine the best concentration of BMP4 at a time of two days on differentiation PGCs from mESC. To differentiate PGCs, embryoid bodies (EBs) from mESCs were cultured in concentrations of 0, 5 and 10 ng/ml BMP4 for two days. Germ cell markers Oct4 (Pou5f1), Stella (Dppa3) and Mvh (Ddx4) were analyzed by flow cytometry, immunocytochemistry and reverse transcriptase polymerase chain reaction (RT-PCR). Flow cytometry data demonstrated most Mvh-positive cells were observed only in the treated groups. Immunocytochemistry of EBs in the treated groups identified cells positive for Mvh. PCR results showed expression of Oct4 in the control group and treated groups. Stella and Mvh were expressed only in the treated groups. Low concentrations of BMP4 during two days had an optimal effect on differentiation of PGCs from mESC.

Important aspect of sperm function such as motility and capacitation appear to be mediated at least partially though hyaloronic acid (HA). Present study investigated effects of different doses of HA on sperm motility and vitality in human. Sperm was obtained from 20 male from IVF clinic in Imam Khomeini Hospital. Sperm motility and vitality in human semen was analyzed according to WHO criteria before and 4 hours after treatment with different doses of HA (0.750, 1000 and 1250 µg/ml). The results showed that in 1000 µg/ml the percent of stage 3 and 4 increased compare to control group. Percent of stage 1 and 2 decreased in group with 1000 µg/ml HA, there was an increase in the percentage of stage 3 and 4 and decrease in percentage of stage 1 and 2 compare to control. In the group treated with 1250 µg/ml stage 1 and 2 increased while stage 3 and 4 decreased. Vitality in all groups decreased except of the group treated with 1000 µg/ml HA. The group with 1250 µg/ml showed significantly decrease in vitality compare to fresh group (P < 0.05). The present study showed that the effects of HA on sperm motility and vitality is dose dependant and 1000 µg/ml HA had the effective role on sperm parameters.

The failure of regeneration after spinal cord injury (SCI) has been attributed to axonal demyelination and neuronal death. Cellular replacement and white matter regeneration are both necessary for SCI repair. In this study, we evaluated the co-transplantation of olfactory ensheathing cells (OEC) and embryonic stem (ES) cell-derived motor neurons (ESMN) on contused SCI. OEC cultured from olfactory nerve rootlets and olfactory bulbs. ESMN was generated by exposing mouse ES cells to retinoic acid and sonic hedgehog. Thirty female rats were used to prepare SCI models in five groups. Control and medium-injected groups was subjected to induce lesion without cell transplantation. OEC or ESMN or both were transplanted into the site of the lesion in other groups. The purity of OEC culture was 95%. Motor neuron progenitor markers (Olig2, Nkx6.1 and Pax6) and motor neuron markers (Isl1, Isl2 and Hb9) were expressed. Histological analysis showed that significantly more (P<0.001) spinal tissue was spared in OEC, ESMN and OEC+ ESMN groups but the OEC+ ESMN group had a significantly greater percentage of spared tissue and myelination than other groups (P< 0.05). The numbers of ESMN in co-transplanted group were significantly higher than ESMN group (P<0.05). A significant (P<0.05) recovery of hindlimb function was observed in rats in the transplanted groups. We found that the co-transplantation of ESMN and OEC into an injured spinal cord has a synergistic effect, promoting neural regeneration, ESMN survival and partial functional recovery.

Free radical formation and oxidative stress might play an important role in the pathogenesis of Parkinson''s disease (PD). In vitro data indicate that neuromelanin (NM) pigment is formed the excess cytosolic catecholamine that is not accumulated into synaptic vesicles via the vesicular monoamine transporter 2 (VMAT2). We designed this study to investigate the neuroprotective effects of vitamin E in the early model of PD. Male rats (n = 40) with unbiased rotational behavior were randomly divided into five groups: sham operated group (SH, n = 8), vehicle-treated SH group (SH + V, n = 8), vitamin E-treated SH group (SH + E, n = 8), vehicle-treated lesion group (L + V, n = 8) and vitamin E-treated lesion group (L + E, n = 8). Unilateral intrastriatal 6-hydroxydopamine (12.5 µl) lesioned rats were treated intramuscularly with α-tocopherol acid succinate (24 I.U/kg, intramuscular [i.m.]) 1 h before surgery and three times per week for 2 month post-surgery. To evaluate the vitamin E pretreatment efficacy, tyrosine hydroxylase (TH) immunoreactivity and immunostaining intensity (ISI) for monoamine transporter 2 were used. TH immunohistochemical analyses showed a reduction of 20 percent in locus coeruleus (LC) cell number of vitamin E pretreated lesioned group but the cell number dropped to 60 percent in the lesioned group. The ISI of the cells was measured for VMAT2 in LC. Lesioned groups: 1) had the lowest VMAT2 ISI of all neurons; 2) There was an inverse relationship between VMAT2 ISI and NM pigment in the locus and 3) Neurons with the highest VMAT2 ISI also had high TH ISI. The data support the hypothesis that repeated i.m. administration of vitamin E exerts a protective effect on the LC neurons in the early model of PD

Routine oocytes cryopreservation remained an elusive technique in the wide ranges of available assisted reproductive technologies. The microtubules of oocytes are vulnerable to cryoprotectants and thermal change during cryopreservation.

The effects of a vitrification protocol were investigated on the spindle and chromosome configurations of mice oocytes cryopreserved at the germinal vesicle stage.

Germinal vesicle with cumulus cells were transferred to vitrification solution which was composed of 30% (v/v) ethylene glycol, 18% Ficoll-70 and 0.3 M Sucrose either by single step or in step-wise way. Following vitrification and in vitro maturation (MII), the matured oocytes were immonostained for meiotic spindles and chromosomes, before visualization using fluorescent microscopy.

A statistically significant increase was observed in the survival and maturation rate in step-wise vitrification (88.96% and 71.23% respectively) compared to single step vitrification (70.6% and 62.42% respectively) (p<0.05). Normal spindle morphology after vitrification-thawing in step-wise vitrification group (77.26%) was higher than single step vitrification group (64.24%) but lower than control group (94.75%) (p<0.05).

The results suggest that vitrification with step-wise procedure on mice germinal vesicle oocytes has positive effects on survival and maturation rate and normal spindle configuration compare with single step vitrification procedure.

Anatomical position is the base for normal posture evaluation. Any deviation from this posture can create problems for an individual. Common faulty postures appear at the head, vertebral column, shoulder girdles, pelvis and other parts of the body. High flexibility of the primary school children’s skeleton increases the chance of faulty postures. Limited athletic activities of Iranian girls can lead to postural deficiencies among them, which can be followed by some irreversible complications. This study evaluated the rate of faulty postures and their risk factors among primary school girls and recommended some procedures for prevention and physical treatment of them. This descriptive study was conducted on all 261 students of Ghaem Motlagh primary school. The subjects ranged between 7 and 11 in age. The data were collected through physical examination carried out by an anatomist and a physiotherapist. These data were analysed using SPSS softwere (ver.6). The finding showed that, 8.8% of the cases had abnormal rotation of the head. In lumbar region 1.5% involved hypolordosis and 6.9% hyperlordosis. Our results showed that there is a significant relationship between scapula winging and dominant hand (p=0.001). According to the results, it is suggested that sport activities under the guidance of a physical therapist, which involve all the body parts should be encouraged especially in girls schools.

The aim of this study was to evaluate changes that occur in motility parameters of progesterone treated mouse spermatozoa during course of hyperactivation. Mouse spermatozoa treated with different doses of progesterone were videotaped after 10 min and 90 min of incubation. For each sperm, one second of movement of the head-midpiece junction was traced from the videotape and for each tracing; seven motility parameters were studied using computer assisted image analysis. For all progesterone treated spermatozoa, motility rate differed significantly from control group after 90 min of incubation. Motility parameters for high doses of progesterone 10 and 100 µg/ml showed hyperactivation occurred during 10 min of incubation. With treatment of 1 µg/ml progesterone, hyperactivated motility pattern of spermatozoa occurred 90 min after incubation similar to the control group showing that low dose of progesterone is unable to induce hyperactivation. In conclusion, progesterone induces hyperactivation in mouse sperm and reduces the motility rate during the time. Iran. Biomed.

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.

Early in the development of many animals, before transcription begins, any change in the pattern of protein synthesis is attributed to a change in the translational activity or stability of mRNA in the egg and early embryo. As a result, translational control is critical for a variety of developmental decisions, including oocyte maturation and initiation of preimplantation development. In this study, using real-time RT-PCR method, we defined the time course of degradation and deadenylation of an oocyte specific gene (c-mos) more precisely and a gene that is re-synthesized after ZGA (cyclin A2). Our data indicate that oocyte-specific transcript, c-mos, degrades rapidly while cyclin A2 mRNA does not and the deadenylation of c-mos mRNA precedes the process of degradation. Our findings suggest that time-dependent elimination of different maternal mRNA is a way for regulation of translation in early development of mouse embryos.

Abstract: This study was designed for evaluation of epiphyseal plate histological changes of femur bones in osteopetrotic op/op mice.In this study 5 osteopetrotic op/op mice which were purchased from the commercial source were used.The animals were killed by overdose of chloroform and their femur bones were extracted. The bones were fixed in 10% formaldehyde and decalcified by HCl (0.6N), and routine histological processing were performed. The sections were stained by H&E methods and studied by conventional light microscopy. The results showed that, proliferative zone (PZ) and especially hypertrophic zone (HZ) were much thickened. In the ossification zone, trabecular bones were irregular and atypical osteoblast cells were observed. The osteoclast cells were not attached to trabecular bones. The bone marrow cavity was restricted and bone marrow cells were poor and scattered. Findings of the present investigation are similar to those reported about epiphyseal plate in osteosclerotic (OC) mice in which epiphyseal plate especially hypertrophic zone was thickened and chondrocytes were not substituted for osteoblasts in calcified cartilage area. Also, osteoclast cells had been inactive or absent in OC mice. For prevention of other complication due to the epiphyseal plate changes in new borne, suitable and punctually treatment protocols such as prescription of Macrophage Colony Stimulating-Factor (MCS-F) could be useful.

Clinical studies have shown that in pathological conditions such as endometriosis and reproductive tract infection (male and female) there is an activation status of macrophages that produce large quantities of nitric oxide (NO) in addition to other effector molecules. Large amounts of NO may have embryotoxic roles and produce infertility. This study was designed to evaluate the effect of different concentrations of NO on mouse pre-implantation embryo development in vitro. Mouse embryos (2-cellstage) were cultured in media containing different concentrations of sodium nitroprusside (SNP), an NO donor, or L-arginine methyl ester (L-NAME), an NO syntase (NOS) inhibitor. At the end of culture, cell apoptosis was studied by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) technique. The results showed that development of preimplantation embryos were inhibited by high concentration of SNP (1 and 10 μM). In contrast, 0.1 μM of SNP stimulated the embryo development. Similarly, the inhibition of NO by NOS inhibitor resulted in the dose-dependentinhibition of embryo development, but the addition of 0.1 μM SNP with L-NAME reversed the inhibitory effect of L-NAME. TUNEL technique showed that high concentration of NO could induce apoptosis in the embryo, but at low concentration, it decreased apoptotic cell death.