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Showing 7 results for Sobhani

Mitra Bakhtiari, Aligholi Sobhani, Mohammad Akbari, Parichehr Pasbakhsh, Mehdi Abbasi, Azim Hedayatpoor, Fardin Amidi , Feridoon Sargolzaei,
Volume 5, Issue 3 (7-2007)

Background: Various approaches have been used in the attempts to improve the quality of frozen–thawed mouse sperms. According to literatures, it seems that hyaluronic acid (HA) has an important role on the permeability and motility of sperms and their interaction with gametes.
Objective: For evaluation of HA supplementation on sperm characteristics and fertilization capability, we investigated the effect of different doses of HA on mouse sperm morphology, motility, vitality and fertilization capability after freezing and thawing.
Materials and Methods: The cauda epididymes was removed from 6 male mice with aseptic method. The sperm samples were frozen in 1.8 ml cryotubes with 18% raffinose and 3% skimmed milk containing cryo-protectant solution. HA at the concentration of 750, 1000 or 1250 µg/ml was supplemented to frozen-thawed sperms. Sperm motility was measured with microscope, and fertilization rate was evaluated after routine IVF by counting the fertilized oocytes. For sperm morphology, papaniclau staining was used while; Eosin B was used for the assessment of sperm viability rate.
Results: HA supplementation (750 µg/ml) improved motility parameters (p < 0.05) and increased the fertility rate (p < 0.05). The effect of 1,000 µg/ml HA was also positive on the sperms. But 1,250 µg/ml HA had negative effect on above mentioned characteristic. On the other hand, none of these doses had any effect on sperm morphology.
Conclusion: The dose of 750 µg/ml of HA has the greatest effect on the motility, vitality and fertility rate of sperms after cryopreservation.

Somaye Khosravi Farsani, Reza Mahmoudi, Mir Abbas Abdolvahhabi, Mehdi Abbasi, Fateme Malek, Aligholi Sobhani,
Volume 5, Issue 5 (7-2007)

Background: Vitrification is assumed to be a promising method to cryopreserve human oocytes but still needs optimizing.
Objective: The aim of this study was to improve the single step and step-wise vitrification effects on maturing mouse GVBD oocytes by ethylene glycol (EG) in conventional straws.
Materials and Methods: Oocytes with compact cumulus cells were cultured for 3hr in TCM199 supplemented with 10% fetal bovine serum (FBS) in 5% CO2 in air. GVBD oocytes were randomly allocated into three groups. (1) Control (non-vitrified group), (2) exposed to single-step vitrification (contained of EG 20%+0.5M sucrose), (3) exposed to step-wise vitrification (2%, 5%, 10%, 20%EG +0.5M sucrose). In vitrification groups,oocytes were thawed and underwent additional 21 hr maturation. Viability of oocytes and maturation to MII stage were analyzed using inverted microscope and additionally by staining of propidium iodide and Hoechst 33342.
Results: All non-vitrified oocytes were viable after 24 hr; however, viability of vitrified samples in single-step group was significantly lower than that of the step-wise and control Groups. Also, the maturation rate in the step-wise group was significantly higher (p < 0.05) compared to single-step.
Conclusion: These results suggest that step-wise vitrification of  GVBD oocytes as compared to single step vitrification  was better in the rate of  survival and in vitro maturation of oocytes.
Reza Mahmoudi, Iraj Amiri, Parichehr Pasbakhsh, Iraj Ragardi Kashani, Mehdi Abbasi, Farid Aboulhasani, Tooba Mehrannia, Aligholi Sobhani,
Volume 6, Issue 5 (7-2008)

Background: 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.
Objective: The effects of a vitrification protocol were investigated on the spindle and chromosome configurations of mice oocytes cryopreserved at the germinal vesicle stage.
Materials and Methods: 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.
Results: 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).
Conclusion: 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.
Seyed Alireza Sobhani, Zahra Etaati, Sepideh Mirani, Paknoosh Saberi, Mahnaz Shiroodi , Hojjat Salmasian, Nadereh Naderi,
Volume 9, Issue 3 (7-2011)

Iron deficiency anemia (IDA) is a common problem in many developing countries. It is still considered the most common nutrition deficiency worldwide. Apart from its direct hematologic importance, IDA affects cellular and humoral immunity and predisposes the host to infections (1).
Pregnant women are highly prone to IDA. Controversial results are reported in studies targeting this group of patients. Tang et al showed a direct association between hemoglobin concentration and the count of CD4+ T-cell lymphocytes, serum levels of IL-2 and IgG, and an inverse association with susceptibility to infection (2). Ironically, Leush et al reported an increase in IgM and IgG in the second and third trimesters of pregnancy in women with IDA (3).
With regard to controversial results and the scarcity of studies focusing on pregnant women, we aimed to enlighten the relation between iron status and some immunological factors include some component of complement system, IgA, IgM, IgG subclasses of immunoglobulins and pro-inflammatory cytokines during the third trimester of pregnancy.
In a descriptive-analytic study participants  were recruited using convenient sampling from  the   women   in  the  third  trimester  of  pregnancy referred to the labor room of gynecology and obstetrics ward of Dr. Shariati Hospital of Bandar Abbas, Iran. Patients with signs and symptoms of thalassemia, infectious diseases or autoimmune diseases were excluded.
IDA were defined with two criteria, hemoglobin concentration of less than 10 mg/dL (its normal range during the third trimesters of pregnancy is 11-14mg/dL) (4) and ferritin less than 40 ng/dL. Patients were categorized into two groups: those with iron deficiency anemia (IDA) and those without this condition (no IDA).
Red cell indices including hemoglobin (Hb) levels, hematocrit (HCT), mean corpuscular volume (MCV), red blood cell distribution width (RDW), and mean corpuscular hemoglobin (MCH), serum iron (SI) and total iron binding capacity (TIBC), concentration of ferritin, C3 and C4 complements and IgA, IgM and IgG subclasses of immunoglobulins were determined. Data was analyzed using SPSS version 11.5 using Student t-test, Pearson’s correlation test and Kolmogorov-Smirnov’s test of normal distribution.
Ninety-two patients were studied. They were aged between 15 and 42 years (mean=25.69±6.2). According to our definition of IDA in pregnancy, 21 patients (22.8%) had IDA.
Our analysis of differences between the two groups in regard to immunologic markers showed that C4 levels are lower in the IDA group (p=0.009) and the levels of C3, IgM, IgG, IgA, IL-1, IL-6 and TNF-α were not statistically different in the two groups .
We noticed that higher levels of serum iron are correlated with higher levels of C3, C4 and IgG1. Due to important properties of IgG1 like complement fixation and opsonic activity, this subclass is dominant antibody to pneumococcal capsular polysaccharides and its deficiency is associated with current infections (5). Taken together noticing key roles of C3, C4 in complement-mediated bacteriolysis, opsonization, facilitated ingestion immune adherence (6) and association of C3, C4 with Iron serum levels found in this study we suggest that decreased level of Iron increases susceptibility of pregnant women to infections like chronic bacterial respiratory infections and recurrent genital herpes (5). Analyzing immunologic parameters differences between the two groups of IDA and no IDA we found that C4 levels are lower in the IDA group but not the levels of C3, IgM, IgG, IgA, IL-1, IL-6 and TNF-α .Our findings about IgM, IgG, IgA are in contradiction to scanty studies in this field. Tang et al (2) about significantly lower level of IgG, CD3+ and CD4+ cells, the ratio of CD4+/CD8+cells, serum IL-2 in second trimester of IDA pregnant woman and Leush et al (3) study showed increase of IgM and IgG in second and third trimesters of IDA groups.
Our findings about non-significant difference in C3 and significant difference in C4 levels in IDA and no IDA groups is  in agreement with Galan et al report about significantly positive correlation of C4 IgA, IgM and Serum ferritin (7). Despite of mounting evidence that TNF, IL-1, and IL-6 cytokines affect hemopoiesis and iron metabolism there was no significant association between IL-1, IL-6 and TNF-α and serum Iron, ferritin, TIBC in our study and inflammatory cytokines were statistically indifferent in the two IDA and no IDA groups. To our knowledge, there are no reports on inflammatory cytokine levels and Iron parameters in pregnant women, but a limited number of reports exploring this field in children and adults.
Bergman et al  which analyzed the in vitro production of IL-1beta, IL-2, IL-6, IL-10, and TNF-α by peripheral blood mononuclear cells from 20 patients with IDA report that  the secretion of the cytokines other than IL-2 did not differ from that of controls (8). In another study there was no difference in serum levels of IL-6 in iron deficiency anemia before and after iron supplementation in children with IDA but in the iron-deficiency group the production of IL-2 was found to be significantly lower than that in controls and became normal after iron supplementation (9). Safuanova et al work about adequate therapy by iron-containing drugs in IDA patients  resulted  in decreased concentrations of IL-1, IL-6, TNF-alpha and INF-gamma and recovering the functional status of the immune system (10). It is possible that discrepancy seen between our results and mentioned reports in non-pregnant patients is a reflection of dramatic change of immune function in pregnancy.
Analyze of the results of our study and similar researches leads us to the conclusion that  unlike extensive immunological changes have been observed in children, IDA has little effect on humoral immunity system of pregnant women, but decrease in  serum iron could predispose them to pyogenic infection  and may predict  increased susceptibility IDA pregnant women to infections.
Reza Mahmoudi, Farzad Rajaei, Iraj Ragardi Kashani, Mehdi Abbasi, Fardin Amidi, Aligholi Sobhani, Iraj Amiri,
Volume 10, Issue 5 (10-2012)

Background: 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.
Objective: The aim of this study was to evaluate viability, maturation and fertilization rate of mouse immature oocytes after single and stepwise vitrification procedure.
Materials and Methods: 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.
Results: 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.
Conclusion: 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.
Maryam Hosseinzadeh Shirzeily, Parichehr Pasbakhsh, Fardin Amidi, Kobra Mehrannia, Aligholi Sobhani ,
Volume 11, Issue 12 (1-2013)

Background: 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.
Objective: 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).
Materials and Methods: 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.
Results: 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).
Conclusion: 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.
Elham Azizi, Mohammad Naji, Maryam Shabani-Nashtaei, Aligholi Sobhani, Atefeh Najafi, Fardin Amidi,
Volume 16, Issue 11 (November 2018)

Background: Vitamin D has multifaceted function in human reproductive physiology. It has been revealed that vitamin D is involved in spermatogenesis, and semen quality can be linked to vitamin D status in men.
Objective: Evaluating the correlation of 25-hydroxy vitamin D (25-OHD) levels in serum with basic and advanced semen parameters and essential determinants of spermatozoa function.
Materials and Methods: Participants were categorized, based on semen parameters, into normozoospermic (NS) and oligoasthenoteratozoospermic (OAT) men. Serum level of 25-OHD was measured. Apoptotic status of spermatozoa, mitochondrial membrane potential and reactive oxygen species content of semen were assessed.
Results: Difference of 25-OHD concentration in serum of NS men versus OAT ones did not meet significance threshold. DNA fragmentation, reactive oxygen species content of semen and mitochondrial membrane potential state revealed significant difference between NS and OAT subjects. There were no significant differences in basic and functional semen parameters when men were stratified based on serum 25-OHD level. Taking both 25-OHD and semen categories (NS and OAT) into consideration did not indicate any significant difference in studied parameters. Total motility of spermatozoa was positively correlated with serum concentration of 25-OHD in all studied subjects. In addition, normal morphology of spermatozoa in NS men revealed a positive and significant correlation with levels of 25-OHD in serum.
Conclusion: Vitamin D may affect motility and morphology of spermatozoa. Lower content of serum vitamin D may affect fertility of men and should be considered in examination of men with abnormal spermogram.

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