Follicle-stimulating hormone receptor gene polymorphism in chronic anovulatory women, with or without polycystic ovary syndrome: a cross-sectional study.

BACKGROUND: Polymorphisms at codons 307 and 680 are the most commonly encountered allelic variants of the follicle-stimulating hormone receptor (FSHR) gene. Studies in Caucasians suggest that certain FSHR variants are more common in women with polycystic ovary syndrome (PCOS) than normal women. The objective of this study was to determine the distribution of FSHR gene polymorphisms at codons 307 and 680 in Thai women with chronic anovulation, without (121 women) and with PCOS (133 women), using 132 known fertile women as controls. METHODS: DNA samples from peripheral blood lymphocytes were extracted and analyzed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The prevalence of Threonine307Threonine (TT), Threonine307Alanine (TA), and Alanine307Alanine (AA) genotypes at codon 307 was 53.0% (95% CI = 44.2-61.7%), 42.4% (95% CI = 34-51.3%), and 4.5% (95% CI = 1.9-10.1%) in controls; 52.6% (95% CI = 43.8-61.3%), 39.8% (95% CI = 31.6-48.7%), and 7.5% (95% CI = 3.9-13.7%) in PCOS women; and 50.4% (95% CI = 42.8-61.2%), 45.4% (95% CI = 34.9-53.1%), and 4.5% (95% CI = 1.5-9.6%) in anovulatory women without PCOS, respectively. The prevalence of Asparagine680Asparagine (NN), Asparagine680Serine (NS), and Serine680Serine (SS) genotypes at codon 680 was 54.5% (95% CI = 45.7-63.2%), 40.9% (95% CI = 32.5-49.8%), and 4.5% (95% CI = 1.9-10.1%) in controls; 51.9% (95% CI = 43.1-60.6%), 44.4% (95% CI = 35.8-53.2%), and 3.8% (95% CI = 1.4-9.0%) in PCOS women; and 47.9% (95% CI = 40.4-58.8%), 47.1% (95% CI = 36.5-54.7%), and 5.0% (95% CI = 2-10.9%) in anovulatory women without PCOS, respectively. The prevalence of FSHR gene polymorphisms at both codons were not statistically different among the three groups. CONCLUSIONS: In Thai women, there was no association between the FSHR gene polymorphism at codons 307 and 680 and chronic anovulation.

Relaxation for perimenopausal and postmenopausal symptoms.

BACKGROUND: Since the time of publication of the Women's Health Initiative (WHI) study, menopausal symptom management has become more complex because of increased awareness of the risks associated with hormone replacement therapy (HRT). Currently, a wide range of management options is available. Some women take prescription drugs, and others use self care strategies, including lifestyle modifications, over-the-counter preparations and complementary and alternative therapies, such as herbal preparations, exercise programmes and relaxation techniques. Relaxation techniques consist of a group of behavioural interventions. They are considered relatively harmless, but their effectiveness in treating vasomotor symptoms and sleep disturbances remains debatable.  OBJECTIVES: To determine the effectiveness of relaxation techniques as treatment for vasomotor symptoms and associated sleep disturbances in perimenopausal and postmenopausal women. SEARCH METHODS: Searches of the following electronic bibliographic databases were performed in February 2014 to identify randomised controlled trials (RCTs): the Cochrane Menstrual Disorders and Subfertility Group Specialised Trials Register; the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, AMED, PsycINFO, Social Science Citation Index and CINAHL. Handsearches of trial registers, relevant journals and published conference abstracts were also performed. SELECTION CRITERIA: RCTs were included if they compared any type of relaxation intervention with no treatment or other treatments (except hormones) for vasomotor symptoms in symptomatic perimenopausal/postmenopausal women. DATA COLLECTION AND ANALYSIS: Two review authors selected studies, assessed quality and extracted data. Included studies were combined, if appropriate, by using a random-effects model to calculate pooled mean differences and 95% confidence intervals. MAIN RESULTS: Four studies were eligible for inclusion (281 participants): Two studies compared relaxation with electroacupuncture or superficial needling, one study compared relaxation with paced respiration or placebo control (α-wave electroencephalographic biofeedback) and one study compared relaxation with no treatment.No evidence was found of a difference between relaxation and acupuncture or superficial needle insertion in the number of hot flushes per 24 hours (mean difference (MD) 0.05, 95% confidence interval (CI) -1.33 to 1.43, two studies, 72 participants, I(2) = 0%; very low-quality evidence). Nor did any evidence suggest a difference between the two interventions in hot flush severity, measured using the Kupperman Index (MD -1.32, 95% CI -5.06 to 2.43, two studies, 72 participants, I(2) = 0%; very low-quality evidence).The other two studies found no clear evidence of a difference in hot flush frequency between relaxation and paced respiration, placebo or no treatment. The data for these comparisons were unsuitable for analysis.None of these studies reported night sweats, sleep disturbances associated with night sweats or adverse effects as an outcome.The main limitations of identified evidence were lack of data, imprecision and failure to report study methods in adequate detail. AUTHORS' CONCLUSIONS: Evidence is insufficient to show the effectiveness of relaxation techniques as treatment for menopausal vasomotor symptoms, or to determine whether this treatment is more effective than no treatment, placebo, acupuncture, superficial needle insertion or paced respiration.

Developmental competence of human embryos derived from in vitro maturation of immature oocytes retrieved during cesarean section.

AIM: To investigate the developmental competence of human embryos that originated from in vitro matured oocytes retrieved during cesarean section. METHODS: Immature oocytes were collected from 95 pregnant women, who underwent cesarean section at Buddhachinaraj Hospital Medical School and consented to participate in the study. Retrieved oocytes were cultured in blastocyst medium supplemented with 75 IU/L of human menopausal gonadotropin. Oocyte maturation was assessed at 30 and 48 h after culture. In vitro matured oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 h. The fertilization, cleavage and blastocyst formation rates were observed. RESULTS: Maturation rate of oocytes after 30 h of culture was 67.9% compared with 13.1% at 48 h (P < 0.0001). Insemination of oocytes in both groups resulted in similar fertilization, cleavage and blastocyst formation rates. CONCLUSION: A large proportion of oocytes retrieved at the time of cesarean section exhibited the capacity to undergo maturation in vitro. They can be fertilized and developed into good-quality blastocyst stage.

Impact of hemostasis methods, electrocoagulation versus suture, in laparoscopic endometriotic cystectomy on the ovarian reserve: a randomized controlled trial.

OBJECTIVE: To evaluate the impact on ovarian reserve between two different methods ofhemostasis after laparoscopic ovarian endometrioma excision. MATERIAL AND METHOD: A randomized controlled study was conducted from January to December 2013 in Thammasat University Hospital, Thailand. Reproductive women, age 18-45years who underwent laparoscopic ovarian cystectomy were randomized in electrocoagulation and suture groups. Clinical baseline data and ovarian reserve outcome (anti-Mullerian hormone (AMH)) were evaluated. RESULTS: Fifty participants were recruited and randomized in two groups. Electrocoagulation and suture groups consisted of 25 participants. Baseline characteristics between 2 groups (age, weight, BMI, height, cyst diameter, duration and estimated blood loss) were not statistically different. There were no significant difference of AMIH between electrocoagulation and suture group atpre-operative (2.90±2.26 vs. 2.52±2.37 ng/ml), 1 week (1.78±1.51 vs. 1.99±1.71 ng/ml), 1 month (1.76±1.50 vs. 2.09±1.62 ng/ml), 3 months (2.09±1.66 vs. 1.96±1.68 ng/ml) and 6 months (2.11±1.84 vs 1.72±1.68 ng/ml), respectively. However mean AMH ofboth groups significantly decreased since the first week of operation. Effect oflaparoscopic ovarian surgery had significantly declined and sustained AMH level until 6 months. CONCLUSION: Laparoscopic cystectomy of ovarian endometrioma has negative impact to ovarian reserve. Either electroco- agulation or suture method had no different effects.

Effects of ovarian endometriotic fluid exposure on fertilization rate of mouse oocytes and subsequent embryo development.

BACKGROUND: Accidental exposure of oocyte/cumulus complex to endometriotic fluid is not uncommon during oocyte retrieval. Only two studies were available on this subject and they gave conflicting results. In this study, we used a mouse model to evaluate the effect of controlled exposure of oocytes to ovarian endometriotic fluid. METHODS: Mouse oocytes/cumulus complexes (n = 862) were divided into 4 groups, and were exposed to endometriotic fluid (group 1), pooled sera from subjects without endometrioma (group 2), phosphate-buffered saline (group 3), and fertilization medium (controls). After five minutes, oocytes were washed and inseminated. Embryo development was observed daily. The quality of hatching blastocysts was assessed by counting the number of inner cell mass (ICM) and trophectoderm (TE) cells. RESULTS: The fertilization, cleavage and blastocyst formation rates in the four groups were not statistically different. The proportions of hatching/hatched blastocysts from fertilized oocytes in groups 1 and 2 were significantly lower than those in group 3 and controls (P = 0.015). Hatching blastocysts from all groups showed no significant difference in the number of ICM and TE cells. CONCLUSIONS: Exposure of mouse oocytes/cumulus complexes to endometriotic fluid had subtle detrimental effects on subsequent blastocyst development. However, one should be cautious in projecting the results of this study to contaminated human oocytes in a clinical setting.

Effects of partial or complete laser-assisted hatching on the hatching of mouse blastocysts and their cell numbers.

BACKGROUND: It is still debatable whether a full-thickness assisted hatching (AH) is better than the partial zona thinning. In this research, we used a mouse model to study the effect of partial and complete laser-AH on the rate of completely hatched blastocyst and their cell numbers. METHODS: In experiment 1, mouse morulae had 0, 1, 2 or 3 full-thickness openings of 10 microns created in the zona pellucida with an infrared laser beam. In the second experiment, 0, 1 and 2 openings of 20 microns were studied. In the third experiment, a full-thickness opening of 20 microns or quarter-thinning of the zonal circumference to a depth of 90% was compared with non-AH controls. RESULTS: No difference in blastocyst formation was found in laser-treated groups and in the controls. In experiment 1, the rate of completely hatched blastocysts was significantly lower than the controls. In experiment 2 when the size of the opening was increased, blastocysts completely hatched at a significantly higher rate than that in the controls. In experiment 3, the rate of completely hatched blastocysts was the highest in the full-thickness group. Cell numbers in completely hatched blastocysts from both AH groups were significantly fewer than those in the controls. CONCLUSIONS: Full-thickness opening resulted in a higher rate of completely hatched blastocysts than quarter zonal-thinning and controls, but the cell numbers were significantly decreased.

Factors affecting pregnancy rates after microsurgical reversal of tubal sterilization.

The aim of this study was to determine the prognostic factors and pregnancy rates after microsurgical reversal of tubal sterilization. Patients undergoing tubal anastomosis from 2001 to 2008 were included. Relevant data were extracted from their medical records. Pregnancy outcomes were ascertained by responses to mailed questionnaires and telephone contact. A total of 98 patients were identified. We found that the mean duration of follow-up was 67 ± 28 months. Fifty-five patients conceived (pregnancy rate 62.5%; 95% confidence interval [CI] 52 to 72.8%). Of these, 50 were intrauterine and 5 were tubal pregnancies. Life-table analysis estimated cumulative pregnancy rates to be 30.7%, 39.8%, 49%, and 53.7% at 6, 12, 18, and 24 months after reversal, respectively. Age at the time of reversal was the only significant prognostic factor multivariate model. We concluded that age of the patient at the operation is the most important prognostic factor.

Effects of crocin supplementation during in vitro culture of intact and half-destroyed four-cell mouse embryos.

OBJECTIVE: We evaluated the effects of crocin supplementation during culture of intact and half-destroyed four-cell mouse embryos. Outcomes measured included rate of cleavage arrest, blastocyst formation, and blastocyst cell number. METHODS: We used laser to create two zonal holes without blastomere destruction in Groups 1 (n=100) and 2 (n=100), and to destroy two of the four blastomeres in Groups 3 (n=150) and 4 (n=150). Embryos were cultured in groups of ten in drops of medium without (Groups 1 and 3) or with 20 µg/ml of crocin supplementation (Groups 2 and 4). RESULTS: Embryos in Groups 1 and 2 had no difference in the rate of cleavage arrest (6.0% vs. 7.0%, respectively; p=0.774) or blastocyst formation (89.0% vs. 86.0%, respectively; p=0.521). Neither was there a difference in the number of cells in the blastocysts (99.6±23.5 vs. 95.6± 8.2, respectively, p=0.83). Half-destroyed embryos cultured in crocin-supplemented medium (Group 4) had a lower rate of cleavage arrest (14.7% vs. 30.0%, p=0.001), and a higher rate of blastocyst formation (51.3% vs. 37.3%, p=0.015), than those in non-supplemented medium (Group 3). In blastocysts derived from half-destroyed embryos, there was no difference in the number of cells in ICM (14.5±3.9 vs. 13.7±2.9, p=0.285), TE (45.2±12.3 vs. 46.0±13.3, p=0.764), or total cells (59.7±12.2 vs. 59.7±14.8, respectively, p=0.990) among the two groups. CONCLUSIONS: Crocin supplementation during in vitro development of impaired embryos improved their development, but had no effect on intact embryos.

Preimplantation genetic diagnosis of alpha-thalassemia-SEA using novel multiplex fluorescent PCR.

PURPOSE: Preimplantation genetic diagnosis (PGD) is an alternative to prenatal diagnosis (PND) giving couples at risk a chance to start a pregnancy with a disease-free baby. This study aimed to develop a new PGD protocol for alpha-thalassemia(-SEA) mutation, the commonest Mendelian disorder. PATIENTS AND METHODS: Multiplex fluorescent PCR was employed for mutation, contamination and linkage analysis. A couple experienced termination of pregnancy following positive PND decided to join the project. RESULTS: Novel primers for alpha-thalassemia(-SEA) mutation amplifying 5 DNA fragments were developed. Two PGD cycles were performed, resulting in an un-affected baby. PND confirmed the heterozygous result. From 24 embryos, 87.5% of affected genotype were of best quality compared to 0% and 18.2% of those with normal and heterozygous, respectively. CONCLUSIONS: A novel PCR protocol for the common alpha-thalassemia(-SEA) mutation is reported. This test should be widely applicable. Interestingly, a potential effect of alpha-thalassemia(-SEA) mutation on preimplantation embryonic development was noticed.

The relationship between reactive oxygen species, DNA fragmentation, and sperm parameters in human sperm using simplified sucrose vitrification with or without triple antioxidant supplementation.

OBJECTIVE: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. METHODS: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. RESULTS: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). CONCLUSION: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.

Assisted reproductive technologies in Thailand: 2001-2007 results generated from the ART Registry, Royal Thai College of Obstetricians and Gynaecologists.

AIM: To present the results of assisted reproductive technology (ART) performed in Thailand during 2001-2007. METHODS: All licensed ART centers are obliged to submit annual reports on the number of patients, cycles, ART techniques and treatment outcomes to the Reproductive Medicine Subcommittee of the Royal Thai College of Obstetricians and Gynaecologists. Data from all centers were aggregated and analyzed retrospectively. RESULTS: Cycles were categorized into fresh and frozen/thawed embryo transfer (FET) cycles. Initiated cycles in the first category for 2001 to 2007 were 2183, 2112, 2780, 2717, 3458, 3579 and 4288, respectively. FET cycles during the same period were 467, 558, 733, 768, 1136, 1210 and 1473, respectively. The average pregnancy rate for in vitro fertilization (IVF) was 28.9% per retrieval (range, 26-32.3%) or 33.8% per transfer (range, 30.7-38.6%). Multiple pregnancies (of which 89.3% were twins) from all treatment procedures during this period were 11.4% (range, 9.2-14.5%). A congenital abnormality was reported in 0.56% of live births. The number of embryos per transfer in IVF decreased from 4.1 to 2.9, with no drop in pregnancy rates. Oocyte insemination by intracytoplasmic sperm injection (ICSI) was utilized more often than standard IVF, while gamete intrafallopian transfer and zygote intrafallopian transfer were almost completely replaced by IVF/ICSI. There was a significant difference in pregnancy rates (P < 0.01) when clinics were classified by cycle volumes (<100, 100-400 and >400 cycles/year). CONCLUSIONS: Despite many limitations, the data provided in this report will help patients, clinicians and policy makers understand the current situation of ART practice in Thailand.

Vitrification of mouse two-cell and blastocyst stage embryos in simplified closed system using either a hemi-straw or a hollow fiber device.

Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8-15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P > .05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P > .05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P < .05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P < .05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners.

Follicular flushing during oocyte retrieval in assisted reproductive techniques.

BACKGROUND: Ultrasound guided transvaginal aspiration of oocytes has replaced other methods of oocyte retrieval for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). However, there is controversy over whether flushing yields a larger number of oocytes and a higher potential for pregnancy than aspiration only. OBJECTIVES: To determine whether follicular aspiration and flushing increases live birth or ongoing pregnancy rates and the number of oocytes over aspiration alone in women undergoing IVF and ICSI. SEARCH STRATEGY: We searched the Menstrual Disorders and Subfertility Group Specialised Register of controlled trials, MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials (CENTRAL), PsycINFO and the citation lists of relevant publications (to April 2010). SELECTION CRITERIA: Randomised controlled trials that compared follicular aspiration and flushing with aspiration alone were included. Trials were excluded if the flushing method comparison was confounded by comparisons of other methods. DATA COLLECTION AND ANALYSIS: Eligible studies were assessed for methodological quality. For dichotomous data, odds ratios (OR) and 95% confidence intervals (CI) were calculated. For continuous data, mean differences were reported. The heterogeneity of the studies was examined by using statistical tests of homogeneity and the I(2) statistic. MAIN RESULTS: No studies reported on the primary outcome of live birth. There was no evidence (3 studies, 164 patients) to suggest an association between follicular aspiration and flushing and ongoing or clinical pregnancy per woman randomised (OR 1.17, 95% CI 0.57 to 2.38). There was no evidence of a difference in adverse events reported between follicular aspiration and flushing and aspiration only. There was no evidence of significant differences in increased oocyte yield per woman randomised (1 study, 44 patients). Without flushing the operative time was significantly shorter, by 3 to 15 minutes (3 studies, P < 0.001) and the dose of pethidine required was significantly less (50 mg versus 100 mg, P < 0.00001). AUTHORS' CONCLUSIONS: There is no evidence that follicular aspiration and flushing is associated with improved clinical or ongoing pregnancy rates, nor an increase in oocyte yield. The operative time is significantly longer and more opiate analgesia is required for pain relief during oocyte retrieval. There is a lack of evidence regarding the effect of follicular aspiration and flushing on live birth rates in the identified data.

Metabolic syndrome and insulin resistance in Thai women with polycystic ovary syndrome.

OBJECTIVE: To determine the prevalence of metabolic syndrome and insulin resistance in Thai women with polycystic ovary syndrome (PCOS). MATERIAL AND METHOD: Oral glucose tolerance tests were performed in 70 PCOS women. Blood was taken for the measurement of fasting insulin, 2-hr postprandial insulin, lipid profiles, testosterone and sex hormone binding globulin RESULTS: The prevalence of metabolic syndrome and insulin resistance was 24.3% and 30.7%, respectively. Diabetes mellitus and impaired glucose tolerance was detected in 11.4% and 31.4%, respectively Homeostatic model assessment (HOMA) was the most sensitive screening test for insulin resistance. PCOS women with metabolic syndrome had significantly higher body mass index, free testosterone index and insulin levels than those without the syndrome. CONCLUSION: There was a high prevalence of metabolic syndrome, insulin resistance, diabetes mellitus and impaired glucose tolerance in Thai PCOS women. A combination of fasting glucose, a 2-hr glucose tolerance test and fasting insulin level effectively identified PCOS women who were at high risk of metabolic syndrome.

Closed-system solid surface vitrification versus slow programmable freezing of mouse 2-cell embryos.

PURPOSE: To compare closed-system solid surface vitrification with slow freezing. METHODS: Mouse 2-cell embryos (n = 348) were divided into vitrification, slow freezing and non-frozen groups. For vitrification, embryos were exposed to 10% ethylene glycol (EG), 10% dimethylsulfoxide (DMSO) and 10% fetal bovine serum (FBS) in phosphate-buffered saline (PBS) for 10 min, then transferred into 17.5% EG, 17.5% DMSO, 0.25 M trehalose and 10% FBS in PBS. They were placed on hemi-straws and inserted into 0.5 ml straws inside a previously cooled aluminum cylinder. Slow freezing was done in straws by the conventional method. RESULTS: Vitrified embryos had significantly higher survival, further cleavage and blastocyst formation rates than those in the slow freezing group (p < 0.001) and were comparable to controls. Blastocysts in the vitrification and control groups had significantly more cells than those in the slow freezing group (p < 0.05). CONCLUSIONS: Closed-system vitrification was more effective than conventional slow freezing.

Gender differences in infertility-related stress and the relationship between stress and social support in Thai infertile couples.

OBJECTIVE: To study infertility-related stress among men and women and to examine its relationship with the level of perceived social support. MATERIAL AND METHOD: The Fertility Problem Inventory (FPI) and the Personal Resource Questionnaire (PRQ) were translated into Thai and used to assess the level of infertility-related stress and perceived social support, respectively, in 238 infertile subjects. RESULTS: The global FPI scores for men and women were 154.2 +/- 18.3 and 154.7 +/- 22.6, respectively (p > 0.05). There was no significant difference in their perceived social support (PRQ scores = 137.8 + 14.0 and 134.0 +/- 16. 7 respectively). A significant negative correlation (r = -0.1894; p < 0.001) existed between global stress and social support in women, but not in men. CONCLUSION: Thai infertile couples experienced a high level of stress. Unlike previous studies from Western countries, there was no gender diference in infertility-related stress.

Developmental competence of in vitro-matured human oocytes obtained from pregnant and non-pregnant women.

OBJECTIVE: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. METHODS: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. RESULTS: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p<0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. CONCLUSION: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.

Rapid freezing versus Cryotop vitrification of mouse two-cell embryos.

OBJECTIVE: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. METHODS: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n=300), a group that underwent Cryotop vitrification (group 2, n=300), and a group that underwent our in-house freezing method (group 3, n=300). RESULTS: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p=0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p=0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p=0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable (88.99±10.44, 88.29±14.79, and 86.42±15.23, respectively; p=0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p<0.001). CONCLUSION: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.

Frequency of Y chromosome microdeletions and chromosomal abnormalities in infertile Thai men with oligozoospermia and azoospermia.

AIM: To investigate the possible causes of oligozoospermia and azoospermia in infertile Thai men, and to find the frequencies of Y chromosome microdeletions and cytogenetic abnormalities in this group. METHODS: From June 2003 to November 2005, 50 azoospermic and 80 oligozoospermic men were enrolled in the study. A detailed history was taken for each man, followed by general and genital examinations. Y chromosome microdeletions were detected by multiplex polymerase chain reaction (PCR) using 11 gene-specific primers that covered all three regions of the azoospermic factor (AZFa, AZFb and AZFc). Fifty men with normal semen analysis were also studied. Karyotyping was done with the standard G- and Q-banding. Serum concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and testosterone were measured by electrochemiluminescence immunoassays (ECLIA). RESULTS: Azoospermia and oligozoospermia could be explained by previous orchitis in 22.3%, former bilateral cryptorchidism in 19.2%, abnormal karyotypes in 4.6% and Y chromosome microdeletions in 3.8% of the subjects. The most frequent deletions were in the AZFc region (50%), followed by AZFb (33%) and AZFbc (17%). No significant difference was detected in hormonal profiles of infertile men, with or without microdeletions. CONCLUSION: The frequencies of Y chromosome microdeletions and cytogenetic abnormalities in oligozoospermic and azoospermic Thai men are comparable with similarly infertile men from other Asian and Western countries.

Septo-optic dysplasia associated with abnormal pubertal development.

Septo-optic dysplasia (SOD) is a congenital anomaly, that is characterized by a triad of optic nerve hypoplasia, structural brain defects, and hypothalamic-pituitary dysfunction. This condition is very rare and it has never been reported in a Thai population. In the present report, the authors described two SOD cases that presented with primary amenorrhea and abnormal pubertal development. Clinical features. Possible etiology of this condition was reviewed