A mild ovarian stimulation strategy in women with poor ovarian reserve undergoing IVF: a multicenter randomized non-inferiority trial.

STUDY QUESTION: In subfertile women with poor ovarian reserve undergoing IVF does a mild ovarian stimulation strategy lead to comparable ongoing pregnancy rates in comparison to a conventional ovarian stimulation strategy? SUMMARY ANSWER: A mild ovarian stimulation strategy in women with poor ovarian reserve undergoing IVF leads to similar ongoing pregnancy rates as a conventional ovarian stimulation strategy. WHAT IS KNOWN ALREADY: Women diagnosed with poor ovarian reserve are treated with a conventional ovarian stimulation strategy consisting of high-dose gonadotropins and pituitary downregulation with a long mid-luteal start GnRH-agonist protocol. Previous studies comparing a conventional strategy with a mild ovarian stimulation strategy consisting of low-dose gonadotropins and pituitary downregulation with a GnRH-antagonist have been under powered and their effectiveness is inconclusive. STUDY DESIGN, SIZE, DURATION: This open label multicenter randomized trial was designed to compare one cycle of a mild ovarian stimulation strategy consisting of low-dose gonadotropins (150 IU FSH) and pituitary downregulation with a GnRH-antagonist to one cycle of a conventional ovarian stimulation strategy consisting of high-dose gonadotropins (450 IU HMG) and pituitary downregulation with a long mid-luteal GnRH-agonist in women of advanced maternal age and/or women with poor ovarian reserve undergoing IVF between May 2011 and April 2014. PARTICIPANTS/MATERIALS, SETTING, METHODS: Couples seeking infertility treatment were eligible if they fulfilled the following inclusion criteria: female age ≥35 years, a raised basal FSH level >10 IU/ml irrespective of age, a low antral follicular count of ≤5 follicles or poor ovarian response or cycle cancellation during a previous IVF cycle irrespective of age. The primary outcome was ongoing pregnancy rate per woman randomized. Analyses were on an intention-to-treat basis. We randomly assigned 195 women to the mild ovarian stimulation strategy and 199 women to the conventional ovarian stimulation strategy. MAIN RESULTS AND THE ROLE OF CHANCE: Ongoing pregnancy rate was 12.8% (25/195) for mild ovarian stimulation versus 13.6% (27/199) for conventional ovarian stimulation leading to a risk ratio of 0.95 (95% CI: 0.57-1.57), representing an absolute difference of -0.7% (95% CI: -7.4 to 5.9). This 95% CI does not extend below the predefined threshold of 10% for inferiority. The duration of ovarian stimulation was significantly lower in the mild ovarian stimulation strategy than in the conventional ovarian stimulation strategy (mean difference -1.2 days, 95% CI: -1.88 to -0.62). Also, a significantly lower amount of gonadotropins was used in the mild simulation strategy, with a mean difference of 3135 IU (95% CI: -3331 to -2940). LIMITATIONS, REASONS FOR CAUTION: A limitation of our study was the lack of data concerning the cryopreservation of surplus embryos, so we are not informed on cumulative pregnancy rates. Another limitation is that we were not able to follow up on the ongoing pregnancies in all centers, so we are not informed on live birth rates. WIDER IMPLICATIONS OF THE FINDINGS: The results are directly applicable in daily clinical practice and may lead to considerable cost savings as high dosages of gonadotropins are not necessary in women with poor ovarian reserve undergoing IVF. A health economic analysis of our data planned to test the hypothesis that mild ovarian stimulation strategy is more cost-effective than the conventional ovarian stimulation strategy is underway. STUDY FUNDING/COMPETING INTERESTS: This study was supported by NUFFIC scholarship (the Netherlands) and STDF short-term fellowship (Egypt). TRIAL REGISTRATION NUMBER: NTR2788 (Trialregister.nl). TRIAL REGISTER DATE: 01 March 2011. DATE OF FIRST PATIENT'S ENROLMENT: May 2011.

Association of interleukin-16 polymorphisms with disease progression and susceptibility in endometriosis.

Interleukin-16 (IL-16) is a multifunctional pro-inflammatory cytokine that was previously found in association with complex disorders, and it is now cleared that this cytokine plays a critical role in regulation of cellular functions such as homoeostasis. Due to the complexity of endometriosis and its resemblance to cancer, we designed present case-control study to determine the effects of genetic polymorphisms of the human IL-16 gene on Iranian women's susceptibility to endometriosis. A total of 126 patients with endometriosis (stages I-IV) and 144 healthy women as control group were recruited to the study. We genotyped four single nucleotide polymorphisms of IL-16 gene (rs11556218 T>G, rs4778889 T>C, rs4072111 C>T and rs1131445 C>T). Genotyping was performed using PCR and restriction fragment length polymorphism. Our results showed that genotype distribution in two exonic polymorphisms including rs11556218 and rs4072111 was significantly different between Endometriosis patients and healthy individuals (P < 0.05). We have also found an association between rs4072111 and rs1131445 with progression to the severe stages (III-IV) of endometriosis (P < 0.05). Finally, we may conclude that IL-16 gene polymorphisms are highly associated with increased risk of endometriosis and could be considered as a susceptibility factor for endometriosis.

Immobilising effect of Ruta graveolens L. on human spermatozoa: coumarin compounds are involved.

This study was designed to find out Ruta graveolens L. functional components, which have immobilisation effect on human spermatozoa for contraceptive use. A five-step fractionation method was used to derive different components from rue aqueous extract by using hexane, chloroform, ethanol, acetone and ultrapure water. Gas chromatography-mass spectrophotometery (GC-MS) of all fractions and the aqueous extract were performed to determine the chemical components. The immobilisation assay and membrane integrity test were also performed with four different coumarins, which were found in GC-MS in a concentration of 10 µm. Hexane, chloroform, acetone and ethanol fractions could significantly decrease motility of sperms within the first and the second hours. Hexane fraction had also significant immediate effect. The aqueous fraction had no effect on sperm motility. Meanwhile, GC-MS revealed that aqueous extract and effective fractions had similar coumarin compounds. We performed the immobilisation assay on four different coumarins, which were found in GC-MS in a concentration of 10 µm. Reduction of sperm motility was only significant for xanthotoxin. In the sperm viability and membrane integrity tests, hexane and ethanolic fractions could impair sperm vitality significantly, in contrast to coumarins. These results indicated that a part of immobilising effect of rue could be due to its coumarins. The possible mechanism could be blocking of spermatozoa potassium channels.

Prevalence of carcinoma in situ in testicular biopsies of infertile Iranian men.

Almost all testicular germ cell tumours are proved to originate from carcinoma in situ cells. Infertility is one of the factors that increase the risk of carcinoma in situ. The reported prevalence for carcinoma in situ from different parts of the world is 0-3.7% in infertile men. This retrospective study was performed to determine the prevalence of carcinoma in situ in Iranian infertile men. We reviewed the testicular biopsies of 1153 infertile men at the pathology department of Avicenna Infertility Center. One hundred and fifty-one cases were suspicious of having carcinoma in situ. Immunohistochemical marker for placental alkaline phosphatase was employed to confirm the diagnosis of carcinoma in situ. Positive results were detected in 7 (0.6%) of 1153 cases (95% CI 0.24%-1.24%), 6 (0.94%) of which (95% CI 0.34%-2.04%) were under the age of 35 years (636 patients were in this age group). This study is the first study in Iran determining the prevalence of carcinoma in situ among the infertile Iranian men; the result is in the range of reports from other countries.

Regeneration of spermatogenesis in a mouse model of azoospermia by follicle-stimulating hormone and oestradiol.

Busulfan is a chemotherapeutic drug that induces sterility, azoospermia and testicular atrophy. To induce degeneration of spermatogenesis, we used different amounts of busulfan. Adult male C57Bl/6 mice were treated with 15, 30 and 45 mg kg(-1) of busulfan. After 5 weeks, animals had daily injections of 7.5 IU human follicle-stimulating hormone (hFSH) and 12.5 µg kg(-1) oestradiol benzoate (EB), separately or simultaneously. After this time, the animals were killed and blood samples were taken through cardiac puncture. Testes were used for histopathology experiments, DNA flow cytometry and RNA extraction for expression of c-kit and cyclin B1 genes. EB unlike FSH has induced stimulatory effects on spermatogenesis, increased the level of serum testosterone 2-fold and caused a 2-fold increase in the number of haploid cells. The result showed that hFSH with EB multiplied EB stimulatory effects on spermatogenesis up to four times. Expression of c-kit and cyclin B1 genes increased in EB and hFSH+EB groups. These findings suggest that EB regulates spermatogonial stem cells via hFSH. hFSH with EB had synergistic effect on regeneration of spermatogenesis.

The micronutrient supplements, zinc sulphate and folic acid, did not ameliorate sperm functional parameters in oligoasthenoteratozoospermic men.

We investigated the effects of folic acid and zinc sulphate supplementation on the improvement of sperm function in subfertile oligoasthenoteratozoospermic (OAT) men. Eighty-three OAT men participated in a 16-week intervention randomised, double-blind clinical trial with daily treatment of folic acid (5 mg day(-1) ) and zinc sulphate (220 mg day(-1) ), or placebo. Before and after treatment, semen and blood samples were obtained for determining sperm concentration, motility, and morphology, sperm viability, sperm mitochondrial function, sperm chromatin status using toluidine blue, aniline blue, acridine orange and chromomycin A3 staining; and semen and blood folate, zinc, B12 , total antioxidant capacity (TAC) and malondialdehyde (MDA) concentrations. Sperm concentration (×10(6)  ml(-1) ) increased in subfertile men receiving the combined treatment of folic acid and zinc sulphate and also in the group receiving only folic acid treatment; however, it was not statistically significant (P = 0.056 and P = 0.05, respectively). Sperm chromatin integrity (%) increased significantly in subfertile men receiving only zinc sulphate treatment (P = 0.048). However, this improvement in sperm quality was not significant after adjusting placebo effect. This study showed that zinc sulphate and folic acid supplementation did not ameliorate sperm quality in infertile men with severely compromised sperm parameters, OAT. Male infertility is a multifactorial disorder, and also nutritional factors play an important role in results of administration of supplementation on sperm parameters. However, these results should be confirmed by multiple studies in larger populations of OAT men.

Effect of Coenzyme Q10 supplementation on antioxidant enzymes activity and oxidative stress of seminal plasma: a double-blind randomised clinical trial.

Low seminal plasma concentrations of coenzyme Q10 (CoQ10) have been correlated with impaired sperm parameters, but the exact mechanism remains of dominating interest. This randomised, placebo-controlled study examined the effect of CoQ10 on catalase, superoxide dismutase (SOD) and F2 -isoprostanes in seminal plasma in infertile men and their relation with CoQ10 concentration. Sixty infertile men with idiopathic oligoasthenoteratozoospermia (OAT) were randomised to receive 200 mg d(-1) of CoQ10 or placebo for 3 months. 47 persons of them completed the study. Semen analysis, anthropometric measurements, diet and physical activity assessment were performed for subjects before and after treatment. Independent and paired t-test, chi-square test and ancova were compared outcomes of supplementation between two groups. CoQ10 levels increased from 44.74 ± 36.47 to 68.17 ± 42.41 ng ml(-1) following supplementation in CoQ10 (P < 0.001). CoQ10 group had higher catalase and SOD activity than the placebo group. There was a significant positive correlation between CoQ10 concentration and normal sperm morphology (P = 0.037), catalase (P = 0.041) and SOD (P < 0.001). Significant difference was shown between the mean of changes in seminal plasma 8-isoprostane in two groups (P = 0.003) after supplementation. Three-month supplementation with CoQ10 in OAT infertile men can attenuate oxidative stress in seminal plasma and improve semen parameters and antioxidant enzymes activity.

Regulation of luteinizing hormone receptor in hippocampal neurons following different long-lasting treatments of castrated adult rats.

The aim of this study was to investigate the effects of different Luteinizing hormone (LH) and steroid hormones levels on LH receptor (LHR) expression in the hippocampal cells. Rats (24 males and 24 females) were assigned to four groups: one control and three experimental [gonadectomy (GDX), gonadectomy + gonadotropin releasing hormone analogue (GDX+GnRHa) and GDX+GnRHa+estradiol (E2) or testosterone (T)] independently for each gender. All experimental rats were gonadectomized; then GnRHa was administrated to GDX+GnRHa group, and GnRHa plus steroid hormone to GDX+GnRHa+E2 or T group in both genders for four-month. LHR mRNA expression and its protein level in hippocampal cells were measured using QRT-PCR and Western blotting. Quantification of mRNA revealed a decrease in LHR transcripts level in GDX+GnRHa group of females. A significant change was observed between GDX groups and GDX+GnRHa+E2 or T versus GDX+GnRHa group in females. High levels of LH decreased significantly the immature isoform of LHR in GDX group compared to control group in both genders, but low LH concentrations in GDX+GnRHa group induced immature LHR isoform production only in females. Therefore increased LH concentration induces production of incomplete LHR transcripts in hippocampal cells and decreases immature LHR at the protein level. This implies that LH decreases the efficiency of translation through either producing non-functional LHR molecules or preventing their translation.

Comparison of intracytoplasmic sperm injection outcomes between spermatozoa retrieved from testicular biopsy and from ejaculation in cryptozoospermic men.

The infrequent presence of spermatozoa in cryptozoospermic men ejaculate is a limiting factor in the treatment of them. Sometimes, this consideration impels us to apply meticulous microscopic search in ejaculate or testicular sperm extraction (TESE) method. The aim of this study was to assess putative effectiveness of sperm origin, ejaculated or testicular, in cryptozoospermia treatment. In this context, were evaluated intracytoplasmic sperm injection (ICSI) outcomes in two parameters including fertilisation rate (2PN) and embryo quality, independently. We compared the outcome in two groups: patients who underwent ejaculate/ICSI and ones who underwent TESE/ICSI process. Nineteen ICSI cycles performed with testicular spermatozoa and the rest of cycles (n = 208) carried out with ejaculated spermatozoa. Result analysis showed similar fertilisation rate between testicular and ejaculated spermatozoa (respectively, 60% versus 68%, P ≥ 0.05). Also, on the other hand, embryo quality did not show significant differences between two groups, except grade A with low significance. With regard to almost equal performance of both methods in results and being invasive of TESE as surgical sperm retrieval method, the use of ejaculated sperm more than testicular sperm should be recommended in patients with cryptozoospermia whenever possible.

Coenzyme Q10 improves seminal oxidative defense but does not affect on semen parameters in idiopathic oligoasthenoteratozoospermia: a randomized double-blind, placebo controlled trial.

BACKGROUND: Several lines of evidence show the implication of oxidative stress in the etiology of male infertility. Recently, the role of coenzyme Q10 (CoQ10) in the prevention and treatment of disease has been intensively probed. However, definitive efficacy studies in oligoasthenoteratozoospermia (OAT) have not been completed yet. AIM: To evaluate the effect of CoQ10 supplementation on semen parameters in idiopathic OAT (iOAT). MATERIAL/SUBJECTS AND METHODS: A double-blind placebo controlled clinical trial was carried out. A total of 47 infertile men with iOAT were randomly assigned to receive 200 mg CoQ10 daily or placebo during a 12- week period. Semen parameters were determined using microscopic evaluation according to World Health Organization guidelines. Lipid peroxidation was assessed by measuring the concentration of plasma malondialdehyde. We evaluated the total antioxidant capacity of seminal plasma. To compare variables between and within the 2 groups we used independent t-test and Paired t-test. RESULTS: The trial showed non-significant changes in semen parameters of CoQ10 group. However, concentrations of thiobarbituric acid-reactive substances were significantly (p<0.05) reduced in serum of treated groups compared with the control. Furthermore, total antioxidant capacity of seminal plasma significantly increased in the CoQ10 group (p<0.05). CONCLUSION: Our results provide further evidence suggesting that CoQ10 supplementation is associated with alleviating oxidative stress, although it does not show any significant effects on sperm concentration, motility and morphology. It may be suggested that CoQ10 could be taken as an adjunct therapy in cases of OAT. Further studies are needed to draw a final conclusion.

Evaluation of semen quality in patients with malignancies referred for sperm banking before cancer treatment.

Different cancer treatments such as chemotherapy and radiotherapy can lead to azoospermia and even sterility for an unknown period. Whether the type of cancer could affect semen quality or not is under debate. In this study, we have reviewed semen parameters of men with cancer who deposited their sperm samples at the Avicenna Research Institute tissue bank before undergoing cytotoxic treatment. This descriptive retrospective study examined 73 cases referred to sperm bank, because of malignancy, prior to initiation of cancer treatments including chemotherapy and radiotherapy. The data recorded were age, marital status, reproductive history, semen analysis reports and cancer history of the patients. Semen samples were analysed according to recommendations of the World Health Organization (1999) before freezing. Results of the analysis showed that 71.2% (52) of patients had oligozoospermia, 93.2% (68) teratozoospermia and 86.3% (63) asthenozoospermia. Different groups of cancer patients did not show any differences in oligozoospermia, teratozoospermia and asthenozoospermia. Impaired spermatogenesis even prior to cancer treatment indicates the importance of fertility preservation. As the majority of patients had suitable specimens for freezing and assisted reproduction, sperm banking is recommended to be performed promptly and before any treatment, especially surgery.

Increased sperm ubiquitination correlates with abnormal chromatin integrity.

Ubiquitin, a 8.5 kDa peptide that marks other proteins for proteasomal degradation, tags defective spermatozoa during epididymal passage and is proposed as a biomarker for sperm quality. The present study was designed to evaluate the relationships between sperm ubiquitination, sperm chromatin integrity and semen parameters. Semen samples from 63 couples were collected and analysed according to World Health Organization criteria. Each sample was evaluated for sperm ubiquitination by the direct immunofluorescence method, using anti-ubiquitin antibodies. Chromatin integrity of the same samples was analysed using acridine orange (AO) and toluidine blue (TB) tests. A positive correlation was found between ubiquitinated spermatozoa and the percentage of spermatozoa with abnormal chromatin (AO: r = 0.58, P < 0.001 and TB: r = 0.48, P < 0.001). Negative correlations were obtained between sperm ubiquitination and: sperm count (r = -0.2, P = 0.048), sperm morphology (r = -0.36, P = 0.003), rapidly progressive motility (r = -0.25, P = 0.044) and slow progressive motility (r = -0.28, P = 0.022). Sperm ubiquitination was positively correlated with the percentage of immotile spermatozoa. These results show that among semen parameters, chromatin abnormality is more closely associated with sperm ubiquitination and further validate sperm ubiquitination as a suitable marker for sperm quality.

Fertilizing capacity of rat spermatozoa is correlated with decline in straight-line velocity measured by continuous computer-aided sperm analysis: epididymal rat spermatozoa from the proximal cauda have a greater fertilizing capacity in vitro than those from the distal cauda or vas deferens.

Rat spermatozoa recovered from different regions of the excurrent ducts of 10 adult males (proximal cauda epididymidis [PC], distal cauda epididymidis [DC], and vas deferens [VD]) were assessed by in vitro fertilization (LVF) using limited sperm numbers, and by continuous evaluation of motility parameters during 5 hours of incubation in vitro with automated computer-aided sperm analysis (CASA). Spermatozoa from the PC region fertilized (68 +/- 6%) a significantly greater (P < or = 0.005) number of oocytes than those from the DC (44 + 5%) or VD (47 +/- 7%). For pooled samples from all three regions, the mean fertilization rate (51 +/- 14%) was less tan for spermatozoa from the PC (P < 0.05) but was not significantly different from spermatozoa from the DC or VD. For each time point and sample, 1,592 +/- 428 sperm tracks were analyzed. CASA was verified by comparison with manual still-frame analysis of video recordings, by repeated analysis of the same or different samples of spermatozoa, and by examination of computer tracks. The coefficients of variation for various motion parameters suggested that the CASA obtained a high degree of precision. There were no significant differences in motility parameters for spermatozoa recovered from equivalent regions of the left or right tract or in motility parameters for spermatozoa from different regions of the tract immediately after recovery. However, during incubation in vitro, spermatozoa from the DC or VD regions exhibited a marked decline in straight-line velocity (VSL) compared with spermatozoa from the PC region. The reduction in VSL (combined values from right and left tract) for DC or VD spermatozoa compared with PC spermatozoa was significant at 2.5 hours of incubation (P < or = 0.05) and highly significant (P < or = 0.005) by the end of the incubation period. Differences in average path velocity (VAP) were also apparent after 4 hours (p < or = 0.05), but no significant differences were observed for measurements of curvilinear velocity (VCL) or lateral bead displacement (ALH). Overall, the decline in VSL over 5 hours was highly correlated (P < or = 0.001) with the outcome of fertilization in vitro. In contrast, initial VSL and changes in VCL of spermatozoa were not correlated with fertilization rate. These results indicate that the in vitro fertilizing capacity of rat spermatozoa is correlated with 1) the decline in straight-line velocity (VSL) as measured by repeated CASA during incubation in vitro and 2) with the site of recovery of mature rat spermatozoa from the distal excurrent duct. It is suggested that the deterioration of the quality of rat spermatozoa in the distal epididymidis and vas deferens during storage may occur sooner than previously realized, and therefore care must be taken when recovering samples for fertility assessment. In keeping with findings in other species, immediate "snapshot" analysis of rat motility was a poor predictor of sperm fertility. In contrast, continuous CASA provided significant information for determining sperm fertilizing capacity and will be a useful technique for reproductive toxicology.

Monoclonal antibodies against ROR1 induce apoptosis of chronic lymphocytic leukemia (CLL) cells.

ROR1 is a receptor tyrosine kinase (RTK) recently identified to be overexpressed at the gene and protein levels in chronic lymphocytic leukemia (CLL). Monoclonal antibodies (MAbs) against RTKs have been successfully applied for therapy of solid tumors. We generated five MAbs against the Ig (n = 1), cysteine-rich (CRD) (n = 2) and kringle (KNG) (n = 2) domains, respectively, of the extracellular part of ROR1. All CLL patients (n = 20) expressed ROR1 on the surface of the leukemic cells. A significantly higher frequency of ROR1 expression was found in patients with progressive versus non-progressive disease, and in those with unmutated versus mutated IgVH genes. All five MAbs alone induced apoptosis in the absence of complement or added effector cells (Annexin-V and MTT, as well as cleavage of poly-(ADP ribose)-polymerase, caspase-8 and caspase-9) of CLL cells but not of normal B cells. Most effective were MAbs against CRD and KNG, significantly superior to rituximab (P < 0.005). Cross-linking of anti-ROR1 MAbs using the F(ab')(2) fragments of anti-Fc antibodies significantly augmented apoptosis. Two of the MAbs induced complement-dependent cytotoxicity (CDC) similar to that of rituximab and one anti-ROR1 MAb (KNG) (IgG1) showed killing activity by antibody-dependent cellular cytotoxicity. The identified ROR1 epitopes may provide a basis for generating human ROR1 MAbs for therapy.

HSV-2 Seroepidemiology and Risk Factors among Iranian Women: A Time to New Thinking.

BACKGROUND: Genital herpes is a common sexually transmitted disease in many developed and developing countries mostly caused by Herpes simplex virus type 2 (HSV-2). This study determines the prevalence of HSV-2 infection between two groups of women with high and low risk behaviors. METHODS: In this seroepidemiologic study, 362 women attending obstetrics and gynecology clinics as low risk group and 156 prisoners and drop in center resident women in Tehran as high risk group were enrolled. HSV infection was identified by serologic tests on blood samples. RESULTS: The prevalence of IgG antibody in high risk group was significantly more than low risk women (26.3% vs. 2.5%). The prevalence of IgM antibody in high risk group was less than low risk group (3.8% vs. 7.1%) but the difference was not statistically significant. In high risk group, there was significant association between positive IgG and anal/oral sex, use of condom, smoking and drug addiction as well as genital pain, burning, itching, ulcer, dysuria, and history of genital infection. In low risk group, association between positive IgM and IgG test results and risky behaviors were not significant. There was significant association between IgM and genital itching, rash, and ulcer. CONCLUSION: Relatively high seroprevalence of anti-HSV-2 IgG and high frequency of genital Herpes among high risk women necessitates regular screening and safe sex education programs. Moreover, risk of acute infection in this group should not be ignored and its distribution in Iranian population should be alarmingly concerned.

Expression profiling of vitamin D receptor in placenta, decidua and ovary of pregnant mice.

OBJECTIVES: The presence of vitamin D receptor (VDR) and the identification of localized vitamin D3 synthesis in placenta and decidua implicate the importance of vitamin D3 in reproductive function. There is, however, no data on the expression profile of VDR in the mouse placenta and endometrium throughout the pregnancy period. STUDY DESIGN: In the present work expression of VDR in reproductive tissues of pregnant mice at different gestational phases has been addressed. Expression of VDR was determined by semi-quantitative RT-PCR, Western blotting and immunohistochemistry. RESULTS: The results showed that VDR mRNA and protein were expressed in decidua, placenta and ovary throughout the pregnancy. VDR gene expression in placenta was significantly elevated in late pregnancy when compared to that of mid pregnancy. Additionally, VDR expression level in decidua rose significantly as pregnancy progressed from early to mid stages. VDR expression in decidua of pregnant mice was higher in comparison to endometrium of non-pregnant mice. Immunohistochemical analysis revealed that VDR protein is consistently expressed by luminal and glandular epithelial cells of decidua, giant cells, glycogen rich cells and labyrinth cells of placenta and by almost all follicular cell types of ovary. Surveying the expression of VDR at the protein level by Western blotting confirmed PCR results. CONCLUSION: It seems that expression of VDR in reproductive organs is finely tuned during pregnancy indicating its eminent role in reproductive biology.

Relationship between sperm chromatin status and ICSI outcome in men with obstructive azoospermia and unexplained infertile normozoospermia.

This study was done to evaluate the effect of sperm source on chromatin integrity and ICSI outcomes. One hundred and thirteen samples containing epididymal aspirates of 57 obstructive azoospermic men and 56-ejaculated semen of normozoospermic men were included in this study. Sperm chromatin status was evaluated by Chromomycin A3 (CMA3), Aniline Blue (AB) and Toluidine Blue (TB). Fertilization rate and embryo quality were recorded. In epididymal group the percentage of sperms stained with AB, CMA3 and TB were significantly higher compared to ejaculate group while fertilization rate (60.6% vs. 74.04%) was significantly lower. However, embryo quality was not significantly different between two groups. In addition, abnormal sperm chromatin condensation and DNA fragmentation were not correlated with fertilization rate and embryo quality. Our results highlight the role of epididymis in sperm maturation and confirm that ICSI using ejaculated sperm is the gold standard for treatment of infertile men.

Indoleamine 2,3-dioxygenase is expressed in the endometrium of cycling mice throughout the oestrous cycle.

Indoleamine 2,3-dioxygenase (IDO), an enzyme responsible for tryptophan catabolism, is thought to be required to prevent the rejection of the allogenic fetus by maternal T cells and to protect against intra- and extra-cellular pathogens. Consequently, we studied the expression of IDO in the endometrium of female Balb/c mice during the oestrous cycle. At each phase, the endometrium was peeled away and the relative expression of IDO mRNA was detected by semi-quantitative RT-PCR. The presence of IDO protein was confirmed in each phase by Western blotting and immunohistochemistry. Our results showed that IDO is expressed in the endometrium of cycling mice during all the phases of oestrous cycle. The expression of IDO was highest at the oestrus and lowest at the dioestrus. By means of Western blotting and immunohistochemistry, we obtained evidence that IDO protein is synthesised in the endometrium of cycling mice throughout the oestrous cycle. In accordance with RT-PCR results, IDO protein was predominant at the oestrus phase. IDO protein was mainly localised in the glandular and luminal epithelial cells. Our results support the concept of IDO providing a mechanism of innate immunity to protect from ascending infections of the female reproductive tract. In addition, considering the fact that mating only occurs during the oestrus phase, the high expression of IDO in this phase is likely to be a mechanism that induces immunological tolerance of the fetus.

Cryoglobulinaemia in hepatitis C-positive patients in Iran.

INTRODUCTION: Cryoglobulins are found in a wide spectrum of disorders but are often transient and without clinical implications. The so-called essential mixed cryoglobulinaemia shows a striking association with hepatitis C virus (HCV) infection (greater than 90 percent). Type II cryoglobulinaemia is the major extrahepatic manifestation of HCV infection. This study aims to investigate the frequency of cryoglobulinaemia in HCV-positive patients in central Iran. METHODS: 50 HCV-positive patients referred to the Shaheed Sadoughi Hospital in Yazd, Iran from May 2004 to December 2005, were included in the study. Their sera were assessed for cryoglobulins. The sera were separated by centrifugation at 37 degrees Celsius and placed in a four degrees Celsius refrigerator in two tubes, one simple and the other, a Wintrobe tube, to see if precipitation occurs during a 48-hour up to a seven-day period. RESULTS: In this study, 50 HCV positive patients were evaluated. Only two patients were female. Mean age was 32 years (ranging from 17 to 52 years). In eight patients, the cryoglobulin test was positive. In seven patients, the test became positive less than 72 hours after sampling. Only one patient who was positive for cryoglobulinaemia, had clinical manifestation of frank vasculitis. CONCLUSION: The prevalence of cryoglobulinaemia in Iran may be lower than other areas, and it may be due to a higher prevalence of cryoglobulinaemia in females and lower prevalence of cryoglobulinaemia in HCV genotype-1a. Most of the cases in our study were male and the more prevalent genotype in Iran is genotype-1a.