OBJECTIVE: Lin28 has been known to control the proliferation and pluripotency of embryonic stem cells. The purpose of this study was to determine the downstream effectors of Lin28 in mouse embryonic stem cells (mESCs) by RNA interference and microarray analysis. METHODS: The control siRNA and Lin28 siRNA (Dharmacon) were transfected into mESCs. Total RNA was prepared from each type of transfected mESC and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to confirm the downregulation of Lin28. The RNAs were labeled and hybridized with an Affymetrix Gene-Chip Mouse Genome 430 2.0 array. The data analysis was accomplished by GenPlex 3.0 software. The expression levels of selected genes were confirmed by quantitative real-time RT-PCR. RESULTS: According to the statistical analysis of the cDNA microarray, a total of 500 genes were altered in Lin28-downregulated mESCs (up-regulated, 384; down-regulated, 116). After differentially expressed gene filtering, 31 genes were selected as candidate genes regulated by Lin28 downregulation. Among them, neuropeptide Y5 receptor and oocyte-specific homeobox 5 genes were significantly upregulated in Lin28-downregulated mESCs. We also showed that the families of neuropeptide Y receptor (Npyr) and oocyte-specific homeobox (Obox) genes were upregulated by downregulation of Lin28. CONCLUSION: Based on the results of this study, we suggest that Lin28 controls the characteristics of mESCs through the regulation of effectors such as the Npyr and Obox families.
Animals ; Chimera ; Down-Regulation ; Embryonic Stem Cells ; Genes, Homeobox ; Genome ; Humans ; Mice ; Neuropeptide Y ; Neuropeptides ; Oligonucleotide Array Sequence Analysis ; Receptors, Neuropeptide Y ; RNA ; RNA Interference ; RNA, Small Interfering ; Statistics as Topic

OBJECTIVE: Sterilization (tubal sterilization and vasectomy) is a widely applied contraceptive method worldwide. Although most studies have described sterilization as a safe method, there are reports of tubal ligation (TL) and vasectomy complications. The aim of this study was to evaluate the effects of TL and vasectomy on the serum oxidative stress, specifically prooxidant-antioxidant balance (PAB) and malondialdehyde (MDA) levels, over time. METHODS: Male and female rats were classified into vasectomy, sham-vasectomy, TL, and sham-TL groups, respectively. The PAB and MDA levels were measured on days 15 and 45 and months 3 and 6 after the intervention. For female rats, blood sampling was performed during the diestrous phase and estradiol and progesterone were also measured. RESULTS: Serum PAB and MDA increased after TL (p<0.05). Vasectomy increased serum MDA remarkably after 45 days, 3 months, and 6 months (p<0.05). After vasectomy, serum PAB also increased although not significantly. Serum estradiol and progesterone decreased remarkably in the TL group compared to the sham group (p<0.05). CONCLUSION: Bilateral TL and vasectomy both increase the serum oxidative stress; however the imbalance after TL was very noticeable. As for the TL, the reduction of serum estrogen levels can be involved in this imbalance. Complications followed by TL or vasectomy could be due to increased levels of oxidants. Thus, prescribing antioxidants during and or after surgery may be a solution.
Adult ; Animals ; Antioxidants ; Contraception ; Estradiol ; Estrogens ; Female ; Humans ; Male ; Malondialdehyde ; Oxidants ; Oxidative Stress ; Progesterone ; Rats ; Salicylamides ; Sterilization ; Sterilization, Tubal ; Vasectomy

OBJECTIVE: This study was undertaken to determine the effect of hypoxia inducible factor (HIF)-1alpha on the cell death, autophagy, and invasion of trophoblasts. METHODS: To understand the effect of HIF-1alpha, we inhibited HIF-1alpha using siRNA under normoxia and hypoxia conditions. Invasion assay and zymography were performed to determine changes in the invasion ability of HIF-1alpha. Western blotting and immunofluorescence were performed to determine some of the signal events involved in apoptosis and autophagy. RESULTS: There was no difference in cell death through the inhibition of HIF-1alpha expression by siRNA; however, the expression of LC3 and autophagosome formation increased. On the other hand, autophagy was increased, and the invasive ability of trophoblast cells decreased according to the inhibition of HIF-1alpha expression by siRNA. These experimental results mean that HIF-1alpha genes regulate the invasive ability of trophoblasts by increasing autophagy. CONCLUSION: This study contributes important data for understanding the mechanism of early pregnancy implantation and the invasive ability of trophoblasts by defining the relationship between the roles of HIF-1alpha and autophagy.
Anoxia ; Apoptosis ; Autophagy ; Blotting, Western ; Cell Death ; Fluorescent Antibody Technique ; Hand ; Placenta ; Pregnancy ; RNA, Small Interfering ; Trophoblasts

OBJECTIVE: To investigate the value of stimulated intrauterine insemination (IUI) in women with unilateral tubal occlusion. METHODS: Superovulation and IUI was performed during 2003-2010 and the medical records were reviewed retrospectively. Thirty-seven infertile women (52 cycles) with unilateral tubal occlusion diagnosed by hysterosalpingography and without other causes of infertility were selected. One-hundred fourteen patients with unexplained infertility served as a control group (182 cycles). The main outcome was the clinical pregnancy rate per cycle. RESULTS: The pregnancy rate per cycle was similar, 17.3% for the unilateral tubal occlusion group and 16.5% for the unexplained infertility group. The rate of miscarriage (11.1% vs. 23.3%) and ectopic pregnancy (11.1% vs. 6.7%) was similar between the two groups. The pregnancy rate was higher in patients with proximal occlusion (25.0%) compared with distal occlusion (13.9%) or unexplained infertility, but not statistically significant. CONCLUSION: Stimulated IUI can be suggested as the initial treatment option in women with unilateral proximal or distal tubal occlusion.
Abortion, Spontaneous ; Fallopian Tube Diseases ; Female ; Humans ; Hysterosalpingography ; Infertility ; Insemination ; Medical Records ; Pregnancy ; Pregnancy Rate ; Pregnancy, Ectopic ; Retrospective Studies ; Sterilization, Tubal ; Superovulation

OBJECTIVE: Previously, we identified that transketolase (Tkt), an important enzyme in the pentose phosphate pathway, is highly expressed at 2 hours of spontaneous maturation in oocytes. Therefore, this study was performed to determine the function of Tkt in meiotic cell cycle regulation, especially at the point of germinal vesicle breakdown (GVBD). METHODS: We evaluated the loss-of-function of Tkt by microinjecting Tkt double-stranded RNAs (dsRNAs) into germinal vesicle-stage oocytes, and the oocytes were cultured in vitro to evaluate phenotypic changes during oocyte maturation. In addition to maturation rates, meiotic spindle and chromosome rearrangements, and changes in expression of other enzymes in the pentose phosphate pathway were determined after Tkt RNA interference (RNAi). RESULTS: Despite the complete and specific knockdown of Tkt expression, GVBD occurred and meiosis was arrested at the metaphase I (MI) stage. The arrested oocytes exhibited spindle loss, chromosomal aggregation, and declined maturation promoting factor and mitogen-activated protein kinase activities. The modified expression of two enzymes in the pentose phosphate pathway, Prps1 and Rbks, after Tkt RNAi and decreased maturation rates were amended when ribose-5-phosphate was supplemented in the culture medium, suggesting that the Tkt and pentose phosphate pathway are important for the maturation process. CONCLUSION: We concluded that Tkt and its associated pentose phosphate pathway play an important role in the MI-MII transition of the oocytes' meiotic cell cycle, but not in the process of GVBD.
Cell Cycle ; Maturation-Promoting Factor ; Meiosis ; Metaphase ; Oocytes ; Pentose Phosphate Pathway ; Protein Kinases ; Ribosemonophosphates ; RNA Interference ; RNA, Double-Stranded ; Transketolase

OBJECTIVE: During IVF, non-transferred embryos are usually selected for cryopreservation on the basis of morphological criteria. This investigation evaluated an application for array comparative genomic hybridization (aCGH) in assessment of surplus embryos prior to cryopreservation. METHODS: First-time IVF patients undergoing elective single embryo transfer and having at least one extra non-transferred embryo suitable for cryopreservation were offered enrollment in the study. Patients were randomized into two groups: Patients in group A (n=55) had embryos assessed first by morphology and then by aCGH, performed on cells obtained from trophectoderm biopsy on post-fertilization day 5. Only euploid embryos were designated for cryopreservation. Patients in group B (n=48) had embryos assessed by morphology alone, with only good morphology embryos considered suitable for cryopreservation. RESULTS: Among biopsied embryos in group A (n=425), euploidy was confirmed in 226 (53.1%). After fresh single embryo transfer, 64 (28.3%) surplus euploid embryos were cryopreserved for 51 patients (92.7%). In group B, 389 good morphology blastocysts were identified and a single top quality blastocyst was selected for fresh transfer. All group B patients (48/48) had at least one blastocyst remaining for cryopreservation. A total of 157 (40.4%) blastocysts were frozen in this group, a significantly larger proportion than was cryopreserved in group A (p=0.017, by chi-squared analysis). CONCLUSION: While aCGH and subsequent frozen embryo transfer are currently used to screen embryos, this is the first investigation to quantify the impact of aCGH specifically on embryo cryopreservation. Incorporation of aCGH screening significantly reduced the total number of cryopreserved blastocysts compared to when suitability for freezing was determined by morphology only. IVF patients should be counseled that the benefits of aCGH screening will likely come at the cost of sharply limiting the number of surplus embryos available for cryopreservation.
Biopsy ; Blastocyst ; Comparative Genomic Hybridization ; Cryopreservation ; Embryo Transfer ; Embryonic Structures ; Fertilization in Vitro ; Freezing ; Humans ; Mass Screening ; Preimplantation Diagnosis ; Single Embryo Transfer

Fertility preservation (FP) is an effort to retain the fertility of cancer patients, and as an emerging discipline, it plays a central role in cancer care. Because of improvement in diagnostic and therapeutic strategies, an increasingly large number of patients are surviving with cancer. FP specialists should make an effort to spread the significance of FP among reproductive women with cancer and provide appropriate education both for associated physicians and for cancer patients who wish to preserve their fertility. Physicians who take part in the initial diagnosis and management of cancer should consider the importance of early referral of young cancer patients to FP specialists and take care of those patients by providing timely information and appropriate counseling. Individualized treatment strategies should be delivered depending on the patient's situation with appropriate team approach.
Counseling ; Female ; Fertility ; Fertility Preservation ; Humans ; Referral and Consultation ; Specialization

Transcription factors govern diverse aspects of cell growth and differentiation as major switches of gene expression. Etv5, a member of the E26 transformation-specific family of transcription factors, has many stories to share when it comes to reproduction. Etv5 deficient mice show complex infertility phenotypes both in males and females. In males, the infertility phenotype exhibited by Etv5 deficiency is sexually dimorphic, and it involves both somatic cells and germ cells. In Etv5-/- female mice, the problem is more complicated by hormonal involvement. This review synthesizes old and new information on this versatile transcription factor-from the inadvertent discovery of its role in the testes to its newly discovered role in maintaining spermatogonial stem cells.
Animals ; Female ; Gene Expression ; Germ Cells ; Humans ; Infertility ; Male ; Mice ; Phenotype ; Reproduction ; Stem Cells ; Testis ; Transcription Factors

OBJECTIVE: The aim of this study was to investigate the effect of insulin sensitizing agents on hormonal and metabolic parameters as well as menstrual patterns in women with polycystic ovary syndrome (PCOS). METHODS: One hundred and twenty-three patients with PCOS were included. Metformin was administered to patients at 1,500 mg or 1,700 mg daily for 3 months. If the patients had no improvement of the menstrual cycle or metformin-related adverse effects developed, the patients changed medication to a daily dose of either 15 mg pioglitazone or up to 45 mg. Then resumption of a regular menstrual cycle or recovery of ovulation was evaluated. Hormonal and metabolic profiles were compared between the response and non-response group to insulin sensitizing agents. RESULTS: One hundred and five patients with PCOS were treated with metformin for 3 months. Forty-eight patients (45.7%) showed improvement of menstrual cycle regularity after 3 months of metformin use, whereas 57 patients (54.3%) had no change. The mean free testosterone measured after 3 months of treatment was significantly lower in metformin responders than in non-responders. The other parameters did not differ between the groups. Of the 23 patients who used pioglitazone for 3 to 6 months, 19 patients (82.6%) showed improvement in their menstrual cycles. CONCLUSION: Metformin treatment seems to be effective for the improvement of menstrual cyclicity irrespective of insulin resistance in women with PCOS. When metformin related adverse effect occurred, pioglitazone would be effective for aiding the resumption of the menstrual cycle.
Female ; Humans ; Insulin ; Insulin Resistance ; Menstrual Cycle ; Metabolome ; Metformin ; Ovulation ; Periodicity ; Polycystic Ovary Syndrome ; Testosterone ; Thiazolidinediones

OBJECTIVE: To evaluate the correlation between serum levels of anti-Mullerian hormone (AMH) and ovarian response to mild stimulation in normoovulatory women and anovulatory women with polycystic ovary syndrome (PCOS). METHODS: Seventy-four cycles of mild stimulation (clomiphene citrate+gonadotropin followed by timed intercourse or intrauterine insemination) performed in normoovulatory women (57 cycles) and anovulatory women with PCOS (17 cycles). Ovarian sensitivity was defined by the number of mature follicles (> or =14 mm) on triggering day per 100 IU of gonadotropin. A correlation between ovarian sensitivity and the baseline serum AMH level (absolute or multiples of the median [MoM] value for each corresponding age) was calculated. Correlation between ovarian response and serum AMH level was evaluated. RESULTS: Ovarian sensitivity to mild stimulation was positively correlated with absolute serum AMH (r=0.535, p<0.001) or AMH-MoM value (r=0.390, p=0.003) in normoovulatory women, but this correlation was not observed in anovulatory women with PCOS (r=0.105, p>0.05, r=-0.265, p>0.05, respectively). CONCLUSION: Ovarian response to mild stimulation is possibly predicted by the serum AMH level in normoovulatory women, but not in anovulatory women with PCOS.
Anti-Mullerian Hormone ; Female ; Gonadotropins ; Humans ; Ovulation Induction ; Polycystic Ovary Syndrome