Effect of manipulating PI3K pathway components on primordial follicle activation and DNA damage response in bovine ovarian follicles in vitro

Developing immature follicles either from fresh or cryopreserved ovarian tissue to attain competent and fertilisable oocytes could provide an option for some patients undergoing fertility preservation to avoid the risk of ovarian micro-metastases following ovarian tissue transplantation. The first stage of an in vitro growth system (IVG) is activation of primordial follicles. Regulation of this process is critical as uncontrolled and precocious growth initiation of primordial follicles during in vitro activation (IVA) has been a major concern. A delicate balance between inhibitory and stimulatory signals is required to achieve activation but this can also be regulated by manipulating key signalling pathways associated with follicle activation. Phosphatase and tensin homolog of chromosome 10 (PTEN), expressed by the oocyte, is a negative regulator of the Phosphoinositide 3-kinase (PI3K) pathway and has been utilised to initiate primordial follicle growth both in vivo and in vitro in a range of species. Pregnancies have been achieved after grafting small ovarian cortical fragments exposed to PI3K/protein kinase B (Akt) activators to reinitiate the growth of residual follicles in the ovarian tissue of premature ovarian insufficiency (POI) patients. However, activating growth in this way may be damaging to the ovarian follicles. PTEN also has a role in maintaining genomic integrity. Its effects on DNA double strand breaks (DSBs) repair capacity has been debatable. Notably, unrepaired DNA damage is related to ovarian ageing. Meiotic errors are also more likely, leading to chromosomal abnormalities and impacting on oocyte quality. Therefore, we hypothesised that inhibiting PTEN to increase the activation of primordial follicles could result in increased DNA damage and compromised DNA repair capacity in oocytes and granulosa cells. This technique may also affect further growth of isolated preantral follicles selected for culture. The overall aim of this thesis was to determine the collective effects of PI3K/PTEN/Akt/mammalian target of rapamycin (mTOR) modulation pathway, either by inhibiting or activating the signals, on primordial follicle activation and DNA damage response (DDR) of bovine ovarian follicles in vitro. These experiments demonstrated that short-term incubation of ovarian cortex with low (1PM) and high dose (10PM) dipotassium bisperoxo (5-hydroxypyridine-2-carboxyl) oxovanadate (V) (bpv(HOpic)), a PTEN inhibitor, increased primordial follicle activation but resulted in a reduction in the proportion of morphologically healthy follicles in the high dose group. In parallel, DNA damage increased with limited DNA repair function. This was observed both in low and high dose. The mTOR signalling pathway is a master regulator of cell growth and metabolism and its inhibition attenuates follicle growth activation. In the second part, the potential benefit of inhibiting PI3K/Akt/mTOR signalling on the regulation of in vitro follicular activation was investigated. The addition of a low dose rapamycin to bpv(HOpic) or rapamycin on its own reduced DNA damage and improved DNA repair capacity of the oocytes. In the last part, experiments were extended to isolated preantral follicles. None of the treatments had an effect on promoting isolated follicle growth and survival. Although DNA repair protein ataxia telangiectasia mutated (ATM) was significantly upregulated in the presence of rapamycin, it appeared that cumulative effects of increased gamma H2A histone family member X (JH2AX) and upregulation of ATM and Rad51 were not sufficient to support follicle growth. Altogether, these data provide unimproved understanding into the regulation of the follicular activation and its relation with DDR, highlighting the significance of getting closer to physiological conditions to maintain follicle integrity. This may be a promising strategy for the derivation of mature oocytes in vitro. However, further investigations at the stage of isolated preantral follicle culture onwards are essential

Age, Low Education and Unemployment are Associated with Intimate Partner Violence among Women

Objective: To describe the prevalence of violence in women visiting Puskesmas Kecamatan Makassar by their partners and their association with socio-demographic factors and gender role perception. Methods: A cross-sectional study using validated WHO questionnaires was conducted on 141 women visiting Puskesmas Makassar in December 2011 to identify presence of physical, sexual or emotional abuse and women’s perception of gender roles. The data was then analyzed using a combination of univariate analysis and Chisquare test. Results: Our study showed that the lifetime prevalence of female abuse in Puskesmas Makassar population was 21.3%, with physical violence constituting 5.7%, sexual violence 6.4% and emotional abuse 17.0% among the subjects. Violence was associated with the wife’s demographic factor (age >35 years, education <9 years) and her partner’s sociodemographic factors (age >35 years and unemployment). Women’s perception of gender role was not significantly associated with prevalence of abuse Conclusion: A woman’s age and her partner’s age of more than 35 years, an education level of less than nine years, and unemployed partner, were all significantly associated with the lifetime prevalence of female abuse, and was not consistent with gender role beliefs. [Indones J Obstet Gynecol 2013; 37-2: 63-70] Keywords: gender role perception, intimate partner violence, Puskesmas Makassa

Telomerase Expression Increased the Risk of Borderline Ovarian Tumors

Objective: To compare the expression of telomerase in benign ovarian tumors and borderline ovarian tumors. Methods: Thirteen samples of paraffin blocks of ovarian borderline tumors and benign ovarian tumors were taken from patients who underwent surgery from January 2006 to December 2011. In all samples, we performed immunohistochemical staining to the paraffin blocks. Semi quantitative determination of the expression of telomerase is done by an Anatomic Pathology specialist and Gynecologist, which already had the same perception about assessing the standardization sample. We used Fisher’s test to analyze the data. Results: There were significant relationship in the moderate expression of telomerase in the nucleus and cytoplasm between benign and borderline ovarian tumors, with an odds ratio of moderate telomerase expression in the cytoplasm and nucleus of 19.3 (95% CI: 1.4 - 943) and 26 (95% CI : 2.3 - 1211). This means that the risk of borderline ovarian tumors in the expression of moderate telomerase in the cytoplasm was 3.19 times compared to the negative expression, whereas in the nucleus it became 26-fold. There is no significant relationship among menopause, age, and telomerase expression in the nucleus and cytoplasm. Conclusion: There are significant differences in the expression of moderate strength telomerase in nucleus and cytoplasm between benign and borderline ovarian tumors. [Indones J Obstet Gynecol 2013; 37-1: 32-40] Keywords: benign ovarian tumor, borderline ovarian tumor, telomerase expressio

Prenatal Diagnosis and Postnatal Management of Meconium Peritonitis

Objective: To present two cases of fetal meconium peritonitis with perforated ileum and without a definite intestinal obstructive lesion. Case report: Two patients who presented prenatally with ultrasound findings of meconium peritonitis and postnatally were found to have perforation of the terminal ileum and meconium peritonitis. In both cases, the diagnosis of meconium peritonitis was suspected prenatally based on the ultrasound findings which were hyperechogenic bowel and abdominal free fluid with increased echogenicity. Both babies were delivered by c-section due to obstetrical indication. After delivery the babies were admitted to the intensive care unit because of a distended abdomen and respiratory distress. An explorative laparotomy revealed perforations of the ileum, and the pediatric surgeon performed adhesiolysis and ileostomy. The infant recovered well after the operation and was fed uneventfully. Conclusions: Echogenic intraabdominal free fluid was the most common ultrasound findings in meconium peritonitis. Early detection of meconium peritonitis was not indicative of poor neonatal outcomes, and selective termination was not necessary, unless indicated for other reasons. [Indones J Obstet Gynecol 2011; 35-4:191-8] Keywords: prenatal diagnosis, postnatal management, meconium peritoniti

Crosstalk between PTEN/PI3K/Akt Signalling and DNA Damage in the Oocyte: Implications for Primordial Follicle Activation, Oocyte Quality and Ageing

The preservation of genome integrity in the mammalian female germline from primordial follicle arrest to activation of growth to oocyte maturation is fundamental to ensure reproductive success. As oocytes are formed before birth and may remain dormant for many years, it is essential that defence mechanisms are monitored and well maintained. The phosphatase and tensin homolog of chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB, Akt) is a major signalling pathway governing primordial follicle recruitment and growth. This pathway also contributes to cell growth, survival and metabolism, and to the maintenance of genomic integrity. Accelerated primordial follicle activation through this pathway may result in a compromised DNA damage response (DDR). Additionally, the distinct DDR mechanisms in oocytes may become less efficient with ageing. This review considers DNA damage surveillance mechanisms and their links to the PTEN/PI3K/Akt signalling pathway, impacting on the DDR during growth activation of primordial follicles, and in ovarian ageing. Targeting DDR mechanisms within oocytes may be of value in developing techniques to protect ovaries against chemotherapy and in advancing clinical approaches to regulate primordial follicle activation

Efek Obat-obat Antiparkinson terhadap Episode Kekantukan Siang Hari pada Pasien Parkinson’s Disease

Masalah tidur terjadi pada 74-98% pada pasien Parkinson’s Disease (PD). Salah satu dari gangguan tidur yang berhubungan dengan PD adalah kekantukan di siang hari yang berlebihan (Execessive daytime sleepiness/EDS). Survai menunjukkan bahwa antara 30 sampai 75% pasien PD melaporkan EDS yang bermakna. Tujuan penelitian in adalah untuk mengetahui sejauh mana obat-obat antiparkinson, tingkat keparahan penyakit, umur subjek, dan durasi penyakit menginduksi terjadinya episode kekantukan siang hari pada pasien PD. Pasien PD yang datang ke Poliklinik Saraf Rumah Cipto Mangunkusumo Jakarta Pusat pada bulan Agustus sampai Oktober 2003, diminta melengkapi Epworth Sleepiness Scale. Terapi yang diterima, usia, skala Hoehn and Yahr, dan durasi penyakit, dicatat dan dianalisis menggunakan Spearman Correlation dan nilai p. Pada penelitian ini tidak terdapat hubungan antara obat-obat antiparkinson, tingkat keparahan penyakit, dan umur subyek dengan kekantukan siang hari pada pasien PD, tetapi terdapat hubungan antara durasi penyakit dengan kekantukan siang hari pada pasien PD

Characteristic and Pregnancy Rate of IVF Patient: a Retrospective Analysis from Two Centres

Introduction: According to recent statistics, infertility in Indonesia reached 21,9%. In-vitro fertilization (IVF) was a reliable treatment option for subfertility couples with success rate of 22-40%. Data from the Indonesia Association for In-Vitro Fertilization (Perfitri) in 2010 showed the country had only 2000 cases, one third of Vietnam’s and half of Thailand’s. The technology’s high cost has been cited as the cause behind the slow development of IVF in Indonesia.Objective: We aim to evaluate the characteristics of patients and their pregnancy rate following in vitro fertilization in infertile couples from two different centres, Yasmin Clinic and Kedoya Clinic.Material and methods: Cohort retrospective was used to analyse outpatient data from two fertility centres. A total of 510 outpatients’ data, 405 in Yasmin Clinic and 105 in Kedoya Clinic, between October 2014 to September 2015 was collected. Patients and in vitro fertilization cycle characteristics from both clinics were evaluated. The pregnancy rate after IVF was compared between the groups. Result: Although maternal age (p=0,032) and paternal age (p=0,017) were statistically significant, they were not clinically significant. There were also no significant differences in the characteristics of in vitro fertilization in both clinics, clinically. However, pregnancy rate was higher in Yasmin Clinic than in Kedoya Clinic, with pregnancy rate of 64,8% and 45,7%, respectively (OR = 2,185; 95%CI 1,420-3,361; p=0,001).Discussion: There were no differences between characteristics of patients and in vitro fertilization cycle in both centres. Nevertheless, pregnancy rate between Yasmin Clinic and Kedoya Clinic were statistically different, suggesting another factors that influence success rate of in vitro fertilization. This research showed that most patients seek medical attention at the age of 35, after 7 years of infertility, suggesting awareness of infertility problem is still low

Characteristic of Poor Responder Patients and the success rate of biochemical pregnancy in the administration of adjuvant recombinant Luteinizing Hormones in Yasmin Clinic, Jakarta

Introduction: Management for poor response patients is still a challenge for clinician engaged in Assisted Reproductive Technology. Various protocols have been proposed to improve the outcome for patients, though no RCT can provide the evidence based of success rate of one protocol among others.Objective: Our purpose is to find the basic data of poor responder patients in Yasmin Clinic, Jakarta and to assess the IVF cycle outcome after the addition of recombinant Luteinizing Hormones.Material and methods: This is a cohort retrospective study taken from medical records of IVF patients at Yasmin Clinic, Cipto Mangunkusumo Hospital between January 2012 to January 2015.Result :  Two hundreds and eighty-one poor response patients were selected for ART during January 2012 to January 2015, were divided into four main group. Control group received gonadotrophin protocol, while group II received additional recombinant Luteinizing hormones (rLH), Group III received additional Growth hormones (GH) and Group IV received additional rLH and GH both.Discussion: Our study demonstrated the follicle count, the oocyte count and the cleavage rate were statistically significant difference between control group and LH group. Where the oocyte count and follicle count were significantly higher, but the cleavage rate with LH showed negative correlation compare with control group.Conclusion : In this study we found that with adjuvant therapy with recombinant Luteinizing Hormones, produced higher follicles count and oocyte count

Expression of CA-125 Level And Neutrophil to Lymphocyte Ratio In Infected And Non-infected Endometrioma

Implantation and growth of endometriosis was determined by immune cell. There were several immunologic cells that promoting implantation and cell proliferation such as macrophages, Natural killer, lymphocyte and monocyte. Infected endometrioma was associated in women with revised American Society for Reproductive Medicine (ASRM) stage III-IV. Neutrophil lymphocyte ratio (NLR) was a simple systemic inflammation response markers. The sensitivity and spesificity CA-125 in predicting endometrioma was very low but it had been used to monitor the progress of endometriosis. Therefore, measuring mean in leucocyte, NLR, PLR and CA-125 level in infected endometrioma was necessary.