The Association between Embryo Development and Chromosomal Results from PGT-A in Women of Advanced Age: A Prospective Cohort Study.

Embryo morphology and morphokinetics have been studied for their association with euploid embryos. However, the results are controversial, especially in the advanced-aged women group, when the risk of aneuploidy increases significantly. This prospective cohort study evaluated the association between embryo development between day-3 cleavage and day-5 blastocyst stages and euploidy rates, determined using preimplantation genetic testing for aneuploidy (PGT-A). Embryos from women aged 35 years and above who underwent intracytoplasmic sperm injections and PGT-A were studied. Day-3 cleavage-stage embryos were evaluated for their cell number, and day-5 blastocyst-stage embryos were evaluated for their morphological grade. Embryo development from day 3 to day 5 was categorized as either good or poor development and evaluated for its association with the PGT-A results. We evaluated 325 embryos from 101 infertile couples. It was found that 55.17% of blastocysts with good development and 29.83% with poor development were euploid. A significant association was found between embryo development and euploidy rates in advanced-aged women (p < 0.001). Also, there were significantly higher rates of euploid embryos with good blastocyst morphological grades, especially blastocyst expansion grades and trophectoderm grades. In conclusion, embryo morphokinetics shows promising results in predicting euploidy in advanced female age.

Clinical utility of combined preimplantation genetic testing methods in couples at risk of passing on beta thalassemia/hemoglobin E disease: A retrospective review from a single center.

Thalassemia and hemoglobinopathy is a group of hereditary blood disorder with diverse clinical manifestation inherited by autosomal recessive manner. The Beta thalassemia/Hemoglobin E disease (HbE/ßthal) causes a variable degree of hemolysis and the most severe form of HbE/ßthal disease develop a lifelong transfusion-dependent anemia. Preimplantation genetic testing (PGT) is an established procedure of embryo genetic analysis to avoid the risk of passing on this particular condition from the carrier parents to their offspring. Preimplantation genetic testing for chromosomal aneuploidy (PGT-A) also facilitates the selection of embryos without chromosomal aberration resulting in the successful embryo implantation rate. Herein, we study the clinical outcome of using combined PGT-M and PGT-A in couples at risk of passing on HbE/ßthal disease. The study was performed from January 2016 to December 2017. PGT-M was developed using short tandem repeat linkage analysis around the beta globin gene cluster and direct mutation testing using primer extension-based mini-sequencing. Thereafter, we recruited 15 couples at risk of passing on HbE/ßthal disease who underwent a combined total of 22 IVF cycles. PGT was performed in 106 embryos with a 3.89% allele drop-out rate. Using combined PGT-M and PGT-A methods, 80% of women obtained satisfactory genetic testing results and were able to undergo embryo transfer within the first two cycles. The successful implantation rate was 64.29%. PGT accuracy was evaluated by prenatal and postnatal genetic confirmation and 100% had a genetic status consistent with PGT results. The overall clinical outcome of successful live birth for couples at risk of producing offspring with HbE/ßthal was 53.33%. Conclusively, combined PGT-M and PGT-A is a useful technology to prevent HbE/ßthal disease in the offspring of recessive carriers.

Breast reconstruction in pregnancy: a case report of multidisciplinary team approach in immediate autologous flap reconstruction for pregnancy-associated breast cancer.

This report presents the results of immediate breast reconstruction with autologous flap in Pregnancy-associated breast cancer (PABC). There was no obstetrics and surgical complications in our report. Immediate breast reconstruction can be performed in PABC after a careful selection. Multidisciplinary team approach is the key in managing these groups of patients.

First successful trial of preimplantation genetic diagnosis for pantothenate kinase-associated neurodegeneration.

PURPOSE: We aim to present a case of a healthy infant born after intracytoplasmic sperm injection-in vitro fertilization (ICSI-IVF) with a preimplantation genetic diagnosis (PGD) for pantothenate kinase-associated neurodegeneration (PKAN) due to PANK2 mutation. METHODS: ICSI-IVF was performed on a Thai couple, 34-year-old female and 33-year-old male, with a family history of PKAN in their first child. Following fertilization, each of the embryos were biopsied in the cleavage stage and subsequently processed for whole-genome amplification. Genetic status of the embryos was diagnosed by linkage analysis and direct mutation testing using primer extension-based mini-sequencing. Comprehensive chromosomal aneuploidy screening was performed using a next-generation sequencing-based strategy. RESULTS: Only a single cycle of ICSI-IVF was processed. There were seven embryos from this couple-two were likely affected, three were likely carriers, one was likely unaffected, and one failed in target genome amplification. Aneuploidy screening was performed before making a decision on embryo transfer, and only one unaffected embryo passed the screening. That embryo was transferred in a frozen thawed cycle, and the pregnancy was successful. The diagnosis was confirmed by amniocentesis, which presented with a result consistent with PGD. At 38 weeks of gestational age, a healthy male baby was born. Postnatal genetic confirmation was also consistent with PGD and the prenatal results. At the age of 24 months, the baby presented with normal growth and development lacking any neurological symptoms. CONCLUSIONS: We report the first successful trial of PGD for PKAN in a developing country using linkage analysis and mini-sequencing in cleavage stage embryos.

Comparison of Cryopreserved Human Sperm between Ultra Rapid Freezing and Slow Programmable Freezing: Effect on Motility, Morphology and DNA Integrity.

BACKGROUND: Cryopreservation of sperm is common methods to preserve male fertility. Sperm freezing, suggest slow programmable freezing caused lower change of sperm morphology than sperm freezing in vapor of liquid nitrogen. Ultra rapid freezing is easy to be worked on, less time, low cost and does not need high experience. OBJECTIVE: To compare the effect on sperm motility, morphology and DNA integrity of post-thawed sperm after ultra rapid freezing and slow programmable freezing methods. MATERIAL AND METHOD: Experimental study at laboratory of infertility unit, Department of Obstetrics and Gynecology, Faculty of Medicine Ramathibodi Hospital. Thirty-seven semen samples with normal semen analysis according to World Health Organization (WHO) 1999 [normal sperm volume ( 2 ml) and normal sperm concentration (≥ 20 x10(6)/ml) and sperm motility (≥ 50%)]. Semen samples were washed. Then each semen sample was divided into six cryovials. Two cryovials, 0.5 ml each, were cryopreserved by slow programmable freezing. Four 0.25 ml containing cryovials, were cryopreserved by ultra rapidfreezing method. After cryopreservationfor 1 month, thawedprocess was carried out at room temperature. Main outcomes are sperm motility was determined by Computer-Assisted Semen Analysis (CASA), sperm morphology was determined by eosin-methylene blue staining and sperm DNA integrity was assessed by TUNEL assay. RESULTS: Sperm motility was reduced significantly by both methods, from 70.4 (9.0)% to 29.1 (12.3)% in slowprogrammable freezing and to 19.7 (9.8)% in ultra rapid freezing (p < 0.05). Sperm motility decreased significantly more by ultra rapid freezing (p < 0.001). The percentage of normal sperm morphology and DNA integrity were also reduced significantly by both methods. However, no significant difference between the two methods was found (p > 0.05). CONCLUSION: Cryopreservation of human sperm for 1 month significantly decreased sperm motility, morphology and DNA integrity in both methods. However sperm motility was decreased more by ultra rapid freezing.

Comparison of desogestrel/ethinyl estradiol plus spironolactone versus cyproterone acetate/ethinyl estradiol in the treatment of polycystic ovary syndrome: a randomized controlled trial.

AIM: To compare the effects of ethinyl estradiol (EE) 30 mcg/desogestrel 150 mcg plus spironolactone 25 mg/day (group A) versus EE 35 mcg/cyproterone acetate 2 mg (group B) on hyperandrogenism and metabolism in PCOS. METHODS: This was a randomized clinical study. Eighteen women in groups A and B received medications for three cycles. Acne score, androgens and metabolic parameters were assessed before and after treatment. RESULTS: One and two women in groups A and B, respectively, were excluded from the study. Both groups had significantly decreased acne score and free androgen index, and increased sex hormone-binding globulin levels. Cholesterol and high-density lipoprotein were significantly increased in group B, and androstenedione was significantly decreased in group A. The regular withdrawal bleeding was obtained in both groups. CONCLUSION: Both regimens had quite similar efficacy on hyperandrogenism after three cycles of therapy and without any changes in metabolic parameters.

Comparative study of nerve fiber density between adenomyosis patients with moderate to severe pain and mild pain.

BACKGROUND: Since some retrospective studies have given inconsistent findings about innervation in adenomyosis, its role in the pain mechanism is still inconclusive. OBJECTIVE: Define the nerve fiber density in adenomyotic tissue as it correlated to pain symptoms. MATERIAL AND METHOD: A cross-sectional study was performed in twenty-five uterine samples from reproductive age women with adenomyosis who underwent either laparotomy or laparoscopic surgery. The nerve fiber density from hysterectomized specimens as measured by immunohistochemistry staining for Protein gene product (PGP) 9.5 and Neurofilament (NF) were compared with the level of pain in the patients as defined by a visual analogue scale and a verbal rating scale. RESULTS: Nerve fibers as detected by PGP9.5 and NF staining in the myometrium were significantly increased in the group of women with adenomyosis experiencing moderate and severe pain as compared to the group experiencing less pain (4 (0, 7) vs. 1.55 (0, 7)/mm2, p-value <0.001, and 6 (3, 10) vs. 0 (0, 4)/mm2, p-value <0.001 respectively). At both phases of the menstrual cycle, the densities of nerve fibers stained with PGP9.5 and NF showed no significant difference. CONCLUSION: These results suggested that the increased of nerve fibers shown in the more severe pain group might play a role in the pathogenesis or symptoms of adenomyosis.

Glucose metabolism in obese and lean adolescents with polycystic ovary syndrome.

Data on glucose metabolism in Asian adolescents with polycystic ovary syndrome (PCOS) are limited. Glucose metabolism assessment using an oral glucose tolerance test (OGTT) in obese and lean Thai adolescents with PCOS, and a comparison between the two groups were done. Thirty-one patients (19 obese, 12 lean) were enrolled. Their median (range) age was 14.9 (11.0-21.0) years. Eighteen patients had abnormal glucose metabolism (13 hyperinsulinemia, 4 impaired glucose tolerance, and 1 diabetes). Compared between obese [median (range) BMI Z-score, 1.6 (1.2-2.6)] and lean [median (range) BMI Z-score, 0.1 (-1.4 to 0.6)] patients, the frequencies of each abnormal OGTT category, areas under the curves of glucose and insulin levels, and insulinogenic index were not different; however, insulin resistance was greater in the obese group. In conclusion, a high proportion of our adolescents with PCOS had abnormal glucose metabolism. Therefore, OGTT should be performed in adolescents with PCOS for the early detection of abnormal glucose metabolism.

The comparison of DNA quantity between full and half volume single cell whole genome amplification by linker-adapter PCR technique.

BACKGROUND: Whole genome amplification (WGA) is a very important step in providing sufficient DNA templates for many high-throughput genetic analyzes. WGA approaches can be subdivided into PCR- or non-PCR-based methods. The PCR amplification category includes PEP-PCR, DOP-PCR and linker-adapter PCR, but only the linker-adapter PCR is suitable for application in preimplantation genetic diagnostic screening because it provides the necessary rapid turnaround time. OBJECTIVE: Evaluate the ability of linker-adapter WGA commercial kits by using half volume compare with full volume of the reagent amplified DNA extracted from single cell fibroblast. MATERIAL AND METHOD: Single cell fibroblast was used based on known genetic profiles. The authors reduced the volume of the reagent and compared the DNA yields and fragmented DNA products with yields and products using the original protocol. RESULTS: Our result did not show a significant difference between the amount of DNA products between full and half volume method (4.72 vs. 4.89 microg, p-value = 0.56). We achieve a slightly different of fragmented length of WGA products, full volume of reagent received slightly longer length than half volume (502.83 vs. 478.30 bp, p-value = 0.19). CONCLUSION: In this study we have shown that the half volume of the reagent of linker-adapter WGA method amplified DNA extracted from single cell fibroblast was comparable DNA yield and DNA fragmented length with the original method. We need further study extrapolate to evaluate the outcome.

Increase accuracy of visual estimation of blood loss from education programme.

OBJECTIVE: To study the increase accuracy of visual estimation of blood loss after an education program. MATERIAL AND METHOD: Seven simulated scenarios with known measured amount of blood were created by using expired packed red cell from blood bank and common surgical materials. Ninety nurses were randomized into two groups. The experimental group attended blood loss estimation course while the control group did not. The percentage of errors in blood loss estimation were calculated and compared between both groups. The main outcome of this study was percentage of nurses who had accurate estimation. We assumed that if the estimated blood volume is within twenty percentage of actual volume it is accurate. RESULTS: There were no difference in age group (p = 0.08), clinical experiences (p = 0.95) and type of work (p = 0.47) between both groups. Educational program significantly increase accuracy in blood loss estimation (p < 0.05) in all seven scenarios. CONCLUSION: Educational program increased the accuracy of visual estimation of blood loss.