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"Latar belakang: Fertilisasi in vitro (FIV) merupakan salah satu metode tata laksana infertilitas yang paling banyak dilakukan di dunia. Kualitas embrio pada FIV sangat dipengaruhi oleh jumlah dan kualitas oosit. Kadar AMH merupakan marka yang rutin diperiksakan pada peserta program FIV. Namun, belum diketahui secara jelas hubungan AMH dengan kualitas dan jumlah oosit.Metode: Penelitian ini merupakan studi analitik observasional retrospektif dengan desain potong lintang pada seluruh peserta program FIV usia 18-45 tahun di Klinik Yasmin, RSCM Kencana, Jakarta, pada periode Januari 2013 hingga Desember 2019. Pengambilan sampel dilakukan secara total sampling. Subjek dengan data tidak lengkap, memiliki etiologi infertilitas berupa sindrom ovarium polikistik, endometriosis, dan faktor sperma dieksklusi dari penelitian. Data kadar AMH, jumlah oosit total, oosit matur, oosit terfertilisasi, dan laju fertilisasi didapatkan oleh pasien.Hasil: Didapatkan sebanyak 692 subjek yang memenuhi kriteria inklusi dan eksklusi. Pada analisis bivariat, didapatkan korelasi kuat antara kadar AMH dengan jumlah oosit total (r = 0,650, p < 0,001), jumlah oosit matur (r = 0,642, p < 0,001), dan jumlah oosit terfertilisasi (r = 0,607, p < 0,001), sedangkan tidak didapatkan korelasi antara kadar AMH dengan laju fertilisasi (r = 0,076, p = 0,049). Setelah dilakukan kontrol terhadap usia dan indeks massa tubuh, didapatkan korelasi antara kadar AMH dengan jumlah oosit total, jumlah oosit matur, jumlah oosit terfertilisasi, dan laju fertilisasi (p < 0,05). Berdasarkan analisis, nilai titik potong kadar AMH serum untuk memprediksi jumlah oosit optimal adalah 1.615 ng/mL (sensitifitas 77%, spesifisitas 77.3%).Simpulan: Kadar AMH serum berkorelasi dengan jumlah oosit total, jumlah oosit matur, jumlah oosit terfertilisasi, dan laju fertilisasi

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Background: In vitro fertilization (FIV) is one of the most widely practiced infertility treatment methods in the world. The quality of embryos in FIV is strongly influenced by the number and quality of oocytes. AMH level is a marker routinely checked on FIV program participants. However, it is not clear the relationship between AMH and the quality and quantity of oocytes.

Method: This study is a retrospective observational analytic study with a cross-sectional design on all FIV program participants aged 18-45 years at the Yasmin Clinic, RSCM Kencana, Jakarta, from January 2013 to December 2019. Sampling was carried out by total sampling. Subjects with incomplete data, having infertility etiology in the form of polycystic ovary syndrome, endometriosis, and sperm factors were excluded from the study. Data on AMH levels, total oocyte count, mature oocytes, fertilized oocytes, and fertilization rate were obtained by the patient.

Result: There were 692 subjects who met the inclusion and exclusion criteria. In the bivariate analysis, there was a strong correlation between AMH levels and the total number of oocytes (r = 0.650, p <0.001), the number of mature oocytes (r = 0.642, p <0.001), and the number of fertilized oocytes (r = 0.607, p <0.001), whereas there was no correlation between AMH levels and fertilization rate (r = 0.076, p = 0.049). After controlling age and body mass index, a correlation was found between AMH levels with total oocyte count, mature oocyte count, fertilized oocyte count, and fertilization rate (p <0.05). Based on the analysis, cut-off of AMH level to predict optimal total oocyte is 1.615 ng/mL (sensitivity 77%, specificity 77.3%).

Conclusion: Serum AMH levels correlate with the total number of oocytes, the number of mature oocytes, the number of fertilized oocytes, and fertilization rate."

"Selama oogenesis, oosit tumbuh dalam ukuran dan memperoleh kompetensi pematangannya. Kompetensi pematangan oosit sangat diperlukan untuk mendukung terjadinya fertilisasi dan perkembangan embrio berkualitas baik. Ketidakmampuan untuk menentukan oosit yang berpotensi menghasilkan embrio yang viable masih menjadi kendala utama pada proses Fertilisasi In Vitro (FIV). Penelitian ini bertujuan mengetahui apakah ukuran diameter oosit bisa menjadi prediktor kualitas oosit dan embrio. Penelitian ini menggunakan desain penelitian prospektif cohort. Metode penelitian untuk pengukuran diameter oosit menggunakan aplikasi Image J, setiap oosit dan sel granulosa dikelompokkan sesuai ukuran diameter oosit, oosit dilakukan fertilisasi dengan teknik ICSI/IMSI kemudian diikuti perkembangan embrio sampai hari ke-5. Pemeriksaan ELISA dipakai untuk mengkonfirmasi kualitas oosit dengan melihat kadar protein Akt dan total Akt pada sel granulosa kumulus oosit, sedangkan untuk mengkonfirmasi kualitas embrio adalah dengan melihat parameter morfokinetik embrio, pencapaian blastokista dan frekuensi abnormalitas pembelahan. Hasil penelitian menunjukkan tidak ada perbedaan signifikan (P>0,05) pada kadar protein p-Akt dan total Akt sel granulosa kumulus antara ketiga kelompok. Selain itu pengamatan morfokinetik embrio menunjukkan tidak ada perbedaan signifikan (P>0.05) kecuali parameter S2, begitu pula dengan pencapaian blastokista serta frekuensi abnormalitas pembelahan kecuali parameter reverse cleavage (RC). Kesimpulan yang didapat dari penelitian ini adalah ukuran diameter oosit tidak mempengaruhi kualitas oosit dan embrio. Ukuran diameter oosit tidak bisa menjadi prediktor kualitas oosit dan embrio, karena tidak ada perbedaan signifikan pada kualitas oosit dan embrio ketiga kelompok. Oosit dengan ukuran diameter kecil memiliki potensi untuk menjadi embrio dengan kualitas baik.

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During oogenesis, oocytes grow in size and acquire maturation competence. Oocyte maturation competence is necessary to support fertilization and the development of good quality embryos. The inability to determine which oocytes have the potential to produce viable embryos remains a major obstacle in the In Vitro Fertilization (IVF). This study aims to determine whether oocyte diameter can be a predictor of oocyte and embryo quality. This study used a prospective cohort research design. The method used for measuring oocyte diameter using Image J application. Each oocyte and cumulus granulosa cells is grouped according to the size of oocyte diameter, oocyte are fertilized with ICSI/IMSI techniques and then followed by embryo development until day 5. ELISA examination is used to confirm oocyte quality by looking at p-Akt and Akt total protein levels in oocyte cumulus granulosa cells, while to confirm embryo quality by looking at the morphokinetic embryo, blastocyst achievement and frequency of cleavage abnormalities. The results showed no significant difference (P>0.05) in p-Akt and Akt total protein levels in cumulus granulosa cells between the three groups. In addition, morphokinetic observation of embryos showed no significant difference (P>0.05) except S2 parameter, as well as the achievement of blastocyst and the frequency of cleavage abnormalities except the reverse cleavage (RC) parameter. The conclusion obtained from this study is that the size of the oocyte diameter does not affect the quality of oocytes and embryos. Oocytes diameter size can not be a predictor of oocyte and embryo quality, because there is no significant difference in the quality of oocytes and embryos of the three groups. Oocytes with a small diameter size have the potential to become embryos with good quality."

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Latar belakang: Suplementasi mikronutrien untuk wanita pada masa prakonsepsi, khususnya zinc dan kalsium, terbukti penting untuk maturasi oosit dan ovulasi. Namun, peran zinc dalam mempromosikan kualitas oosit dan potensi perkembangannya belum diketahui secara jelas. GDF9, anggota superfamili TGF b yang disekresikan dari oosit selama proses folikulogenesis, terbukti dapat menjadi biomarker maturasi nuklear oosit dan kualitas embrio. Tujuan: Studi potong lintang ini bertujuan untuk mengetahui apakah ada hubungan antara kadar zinc dan kalsium dalam serum dan cairan folikel dengan ekspresi GDF9 terhadap maturasi oosit. Selain itu, penelitian ini juga bertujuan untuk mengetahui apakah kadar zinc dan kalsium dalam serum dapat mewakili kadar zinc dan kalsium dalam cairan folikel. Metode: Studi ini dilakukan pada 25 subjek penelitian yang menjalani program fertilisasi in vitro di Poliklinik Yasmin RSUPN Cipto Mangunkusumo (RSCM) Kencana. Darah vena sebanyak 6 cc diambil pada hari ovum pick up (OPU) dan kemudian dianalisis di laboratorium untuk mengetahui kadar zinc, kalsium, dan protein GDF9. Cairan intrafolikuler dan sel granulosa juga akan diambil dan diperiksa kadar zinc dan kalsium dari cairan intrafolikuler serta ekspresi mRNA GDF9 dari sel granulosa. Hasil:  Dari 25 subjek penelitian, 12 subjek (48%) di antaranya dikategorikan ke dalam kelompok angka maturasi oosit baik (berdasarkan indikator oosit matur dari konsensus Vienna) serta 13 (52%) sisanya dikategorikan ke dalam kelompok angka maturasi oosit buruk. Dari uji korelasi antara kadar zinc dan kalsium dalam serum dengan cairan folikel, kadar zinc folikel terbukti berkorelasi secara signifikan dengan kadar zinc serum (p = 0,019). Kadar GDF9 serum juga terbukti berkorelasi secara signifikan dengan ekspresi GDF9 (p = 0,047). Tidak didapatkan korelasi yang bermakna antara kadar zinc dan kalsium serum dengan kadar GDF9 serum serta ekspresi mRNA GDF9 dari sel granulosa terhadap angka maturasi oosit (p > 0,05). Kesimpulan: Kadar GDF9 serum dapat menjadi pengganti biomarker untuk kualitas oosit. Tidak didapatkan hubungan antara kadar zinc dan kalsium dalam serum atau cairan folikel terhadap kadar GDF9 serum atau ekspresi mRNA GDF9 dari sel granulosa terhadap angka maturasi oosit.

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Background:  Micronutrient supplementation for women during preconception, especially zinc and calcium, is critical for oocyte maturation and ovulation. However, the role of Zinc in promoting quality of oocytes has not yet been elucidated. GDF9, one of oocyte sereting factor, has been proven to be a biomarker of maturation of nuclear oocyte and quality of embryo. Aim: to investigate any relationship between zinc and calcium levels in serum and follicular fluid and GDF9 expression towards maturation of oocytes. In addition, this study also aimed to determine whether zinc and calcium levels in serum could represent zinc and calcium levels in follicular fluid. Method: This cross-sectional study was conducted on 25 subjects who underwent IVF programs at the Yasmin Policlinic, RSCM Kencana. Six mililiters of venous blood was taken on the day of the ovum pick up (OPU) and then analyzed in the laboratory to determine the levels of zinc, calcium, and protein GDF9. In addition to venous blood, intrafollicular fluid and granulosa cells will also be taken and examined zinc and calcium levels from intrafollicular fluid and GDF9 mRNA expression from granulosa cells. Result: 12 (48%) out of 25 subjects were categorized into high oocyte maturation rate (based on Vienna consensus on oocyte maturation rate), and the other 13 (52%) were categorized into low oocyte maturation rate. Follicular zinc levels were significantly correlated with serum zinc levels (p = 0,019). Serum GDF9 levels were also significantly correlated with expressions of GDF9 mRNA (p = 0,047). No significant correlation was found between serum levels of zinc and calcium and serum GDF9 levels or GDF9 mRNA expression towards maturation of oocytes (p > 0,05). Conclusion: Serum GDF9 might substitute for follicular GDF9 as a biomarker of oocyte quality. There is no relationship between serum or follicular zinc/calcium levels and serum GDF9 levels or GDF9 mRNA expression from granulosa cells towards oocyte maturation rates.

 

 

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"Tujuan : Untuk mengetahui hubungan antara GDF-9 dan BMP-15 Serum dan cairan folikel, untuk memprediksi kualitas oosit pada wanita yang menjalani siklus fertilisasi in vitro (FIV). Metode : Studi  Potong Lintang dengan jumlah sampel penelitian sebanyak 30 darah sérum dan 30 sampel cairan folikel yang diambil saat petik telur (OPU), dilakukan pemeriksaan GDF-9 dan BMP-15 menggunakan kit ELISA. Dilakukan analisa dengan uji korelasi Pearson dan Spearman  untuk melakukan analisa hubungan antara GDF-9 dan BMP-15 serum dan cairan folikel dengan parameter-parameter kualitas oosit seperti laju maturasi dan laju fertilisasi. Hasil : rerata usia subyek penelitiann adalah 35,0(26,0-39,0) tahun. Kadar GDF-9 cairan folikel adalah 163,0 pg/ml (48,0-537) dan kadar GDF-9 serum 260,33 pg.ml ± 121,82) sedangkan kadar rerata BMP-15 cairan folikel adalah 58,30 pg/ml ± 31,54 dan kadar BMP-15 serum 74,20 pg/ml (1,0-610). Tidak terdapat hubungan yang bermakna antara kadar GDF-9 serum dan GDF-9 cairan folikel (p =0,245) sedangkan antara BMP 15 serum dan BMP-15 cairan folikel  terdapat hubungan bermakna (p =0,001). Simpulan : Terdapat sebaran yang tidak normal kadar GDF-9 serum dan cairan folikel, keduanya tidak berkorelasi. Terdapat korelasi positif kadar BMP-15 serum dan BMP-15 cairan folikel. Terdapat  korelasi yang kuat. antara BMP-15 serum dengan laju maturasi. Tidak terdapat korelasi antara kadar GDF-9 serum, GDF-9 cairan folikel dan BMP-15 cairan folikel dengan laju maturasi dan laju fertilisasi. GDF-9 serum-cairan folikel dan BMP-15 serum-cairan folikel tidak dapat memprediksi kualitas oosit.

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Aim : To determine the relationship between GDF-9 and BMP-15 serum within follicular fluid in order to predict the quality of oocytes in women undergoing In Vitro Fertilisation (IVF). Method : We collected 30 samples of blood serum and 30 samples of follicular fluid on the day of ovum pickup (OPU), and we examined GDF-9 and BMP-15 using ELISA kits. Analysis by Pearson and a partial-correlation was conducted to analyse the correlation between the concentration of GDF-9 and BMP-15 in serum and follicular fluid with general physiological parameters, such as maturation rates and fertilisation rates.Results : The mean age of the subjects was 35,0 (26,0-39,0) years. The level of GDF-9 in the follicular fluid was 163,0 pg/ml (48,0-537), and the level in the serum was 260.33 pg/ml±121,82. The level of BMP-15 in the follicular fluid was 58,30 pg/ml ± 31,54, and the level in the serum was 74,20 pg/ml (1,0-610). There was no correlation between GDF-9 serum and follicular fluid (P = 0.245) but any correlation  between BMP-15 and follicular fluid (p = 0.001). Conclusion : There was an abnormal distribution of GDF-9 serum and follicular fluid levels, both of them not correlate. There was a positive correlation between BMP-15 serum and BMP-15 follicular fluid. There was strong correlation between BMP-15 serum and maturation rates. No correlation between levels of GDF-9 serum-follicular fluid, and BMP-9 follicular fluid with maturation and fertilization rates. GDF-9 serum-follicular fluid, BMP-15 serum-follicular fluid are not able to predict the quality of oocytes."

"Pendahuluan: Angka keberhasilan FIV di Indonesia sekitar 32-35%. Salah satu penyebab pencapaian yang rendah ini adalah mutu oosit yang dinilai secara mi­kros­kopik pada saat panen oosit. Dari sekian banyak faktor yang berperan dalam pembentukan oosit matang bermutu baik diduga yang paling menentukan per­olehan oosit ma­tang, jumlah fertilisasi, dan jumlah embrio yang dipindahkan ke ute­rus pada FIV adalah AMH, inhibin-B, IGF-2 dan nisbahnya. Bahan dan metoda: Kajian analitik potong-lintang pengukuran berulang dilaku­kan pada bu­lan September 2013-Agustus 2014 di Rumah Sakit Anak dan Bunda Harapan Kita, Jakarta. Sebanyak 38 pasien berumur 26-42 tahun yang mengikuti program FIV diukur kadar AMH, inhibin-B, IGF-2 saat basal, pencetus, panen oosit dalam serum dan dalam zalir folikel. Analisis regresi linear digunakan untuk memperoleh faktor penduga jumlah perolehan oosit matang, jumlah fertilisasi, dan jumlah embrio yang dipindahkan.Hasil: Parameter penduga perolehan oosit matang adalah inhibin-B serum panen oosit dan folikel antral basal (FAB) total. Parameter penduga jumlah fertilisasi adalah FAB to­tal, nisbah inhibin-B pencetus terhadap inhibin-B basal, dan nisbah IGF-2 pen­cetus terhadap inhibin-B pencetus. Parameter penduga jumlah embrio yang dipin­dahkan adalah FAB total, inhibin-B panen oosit, dan nisbah inhibin-B panen oosit terhadap inhi­bin-B pencetus.Pada analisis bivariat area under curve (AUC) terbesar (77,4%) ditemukan pada titik-potong in­hi­bin-B serum panen oosit. Kadar inhibin-B panen oosit yang lebih tinggi dari 131,16 ng/ mL adalah akurat untuk menetapkan kematangan oosit de­ngan ke­pe­­kaan (sensiti­vitas) 84% dan kekhasan (spesifisitas) 69,2%.Simpulan: Inhibin-B serum saat panen oosit berhubungan dengan pembentukan oo­sit matang dan normal sehingga dapat dijadikan parameter penduga perolehan oosit ma­tang dan jum­lah em­brio yang terbentuk. Ditemukan parameter-parameter baru, yaitu (1) nisbah inhibin-B pen­ce­tus terhadap inhibin-B basal serum, dan nisbah IGF-2 pen­cetus terhadap in­hibin-B pencetus serum untuk menduga jumlah fertili­sasi; (2) nisbah inhibin-B pa­nen terhadap inhibin-B pen­­cetus serum untuk menduga jumlah em­brio yang dipindahkan ke uterus pada FIV.

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Background: The success rate of IVF in Indonesia was 30-35%. This low rate was caused by the microscopically evaluated oocyte quality that was obtained by ovum pick-up (OPU). The determinatively contributing factors for the formation of good quality mature oocytes, which are considered to be used as predictive parameter for mature oocytes recovery, number of fertilization, and number transferrable embryos in IVF, are AMH, inhibin-B, IGF-2, and their ratios. Therefore, the study to determine the correlation of those factors with the formation of ferti­li­zable mature oocyte in IVF program is necessary.

Materials and methods: An analytic cross-sectional repeated measurements study was conducted from September 2013 until August 2014 at Harapan Kita Mother and Chil­d Hospital, Jakarta. There were 38 patients aged between 26-42 years who par­ticipated in the IVF program; all of them underwent measurement for serum AMH, inhibin-B, and IGF-2 levels at basal, trigger, and OPU times. Predictive parameters for the number of mature oocytes, fertilizable oocytes, number of embryos transferred were analysed using linear regression.

Results: Predictive parameter for the number of mature oocytes are inhibin-B at OPU and total basal antral follicle (BAF) count. Predictive factors for the number of fertili­za­tion are total BAF count, the ratio of inhibin-B at triggering to inhibin-B at basal ti­mes. Predic­ti­ve factors for the number of embryos transferred are total BAF, inhibin-B at OPU, and the ratio of inhibin-B at OPU to inhibin-B at triggering time.

Using bivariate analysis, at the largest area under the curve (AUC) which was as high as 77.4%, the cut-off point of serum inhibin-B at OPU was found. The serum inhibin-B level at OPU higher than 131.16 ng/mL is accurate for determining the oocyte ma­turity (84% sensitivity and 69.2% spe­cificity).

Conclusions: Serum inhibin-B at OPU correlates with the formation of both mature and normal oocytes, thus it can be used as a predictor for the number of mature oo­cytes recovered and the number of embryos transferred. New parameters are found, those are: (1) the ratio of inhibin-B at triggering to inhi­bin-B in serum at basal times; and the ratio of IGF-2 at triggering to inhibin-B in serum at triggering times to pre­dict the number of fer­tilization; (2) the ratio of inhibin-B at OPU to inhibin-B in se­rum at triggering times to pre­dict the number of embryos transferred."

"Latar Belakang: Growth Differentiation Factor 9 (GDF9) adalah salah satu anggota dari superfamili TGF-β yang merupakan salah satu oocyte-secreted factor (OSF). GDF9 memainkan peran penting dalam folikulogenesis ovarium, kompetensi perkembangan oosit, serta sebagai molekul esensial yang mengontrol berbagai proses sel granulosa dan laju ovulasi. GDF9 merupakan OSF yang memainkan peran penting dalam menjaga fertilitas wanita. Penelitian ini bertujuan untuk mengetahui apakah ekspresi GDF9 berkorelasi dengan kualitas oosit dan laju fertilisasi pada pasien fertilisasi in vitro.Tujuan: Mengetahui korelasi ekspresi GDF-9 dengan kualitas oosit dan laju fertilisasi pada pasien Fertilisasi In VitroMetode: Penelitian ini adalah penelitian potong lintang yang dilaksanakan di Klinik Yasmin, Rumah Sakit dr. Cipto Mangunkusumo, Jakarta sejak bulan Juli 2019 sampai bulan Juli 2020. Terdapat 26 wanita berusia 25-40 tahun pasien Fertilisasi In Vitro tanpa penyakit endometriosis maupun Sindrom Ovarium Polikistik yang menjadi subjek penelitian ini. Cairan folikel pasien diambil, kemudian dilakukan pengukuran ekspresi gen GDF9. Setelah itu, dilakukan evaluasi pada data ekspresi gen GDF9 serta diuji korelasinya dengan kualitas oosit dan laju fertilisasi pasien menggunakan aplikasi SPSS.Hasil: Sebanyak 26 pasien IVF berpartisipasi dalam penelitian ini. Median jumlah ekspresi gen GDF9 adalah sebanyak 2.47 x 10-5 ng/μl dengan median kualitas oosit dan laju fertilisasi pasien IVF sebesar 3.00 dan 0.60. Berdasarkan hasil uji korelasi, terdapat korelasi negatif antara ekspresi GDF9 terhadap laju fertilisasi dengan kekuatan korelasi sedang (r = -0.443, p = 0.012). Sementara, ditemukan korelasi tidak bermakna antara ekspresi GDF9 terhadap kualitas oosit (r = -0.306, p = 0.064).Kesimpulan: Terdapat korelasi negatif dengan kekuatan korelasi sedang antara ekspresi GDF9 dengan laju fertilisasi, sementara hubungan ekspresi GDF-9 dengan kualitas oosit memiliki korelasi tidak bermakna.

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Background: Growth Differentiation Factor 9 (GDF9) is a member of the TGF-β superfamily which is one of the oocyte-secreted factor (OSF). GDF9 plays an important role in ovarian folliculogenesis, the competence of oocyte development, as well as an essential molecule that controls various granulosa cells process and the rate of ovulation. GDF9 is an OSF which plays an important role in maintaining female fertility. This study is conducted to see the correlation between the expression of GDF9 gene with the oocyte quality and the fertilization rate in the IVF patients.

Aim: To find out the correlation between the expression of GDF9 with the oocyte quality and the rate of fertilization of IVF Patients

Methods: This study is a cross-sectional study which was conducted at Klinik Yasmin dr. Cipto Mangunkusumo General Hospital, Jakarta since July 2019 to July 2020. 26 IVF patients aged 25-40 years without the conditions of endometriosis or Polycystic Ovary Syndrome (PCOS) participated in this study. Follicular fluids are taken from the patients and the GDF9 gene expression were measured. The data were then evaluated for its correlation with the oocyte quality and the fertilization rate using the SPSS application.

Results: Abour 26 IVF patients were participated in this study. The median amount of GDF9 gene expression was 2.47 x 10-5 ng/μl. The median of the oocyte quality and the fertilization rate was 3.00 and 0.60. According to the correlation test, there is a negative correlation between the expression of GDF9 gene and the fertilization rate with a moderate statistical correlation (r = -0.443, p = 0.012). Meanwhile, the expression of GDF9 gene has no significance correlation with the oocyte quality (r = -0.306, p = 0.064).

Conclusion: This study showed a negative correlation between the expression of GDF9 gene and the fertilizatio rate, while the relation between the GDF9 expression and the occyte quality has no significance correlation."

"LATAR BELAKANG: Anti Mullerian Hormon (AMH) adalah anggota dari golongan Transforming Growth Factor-β yang berperan dalam pengaturan folikuligenesis pada reproduksi wanita. Peningkatan kadar AMH 2 sampai 3 kali dijumpai pada pasien SOPK (Sindrom Ovarium Polikistik) daripada wanita dengan ovulasi normal. Pada penelitian ini dideteksi varian sekuen disepanjang daerah promoter gen AMH. Adanya variasi promoter gen AMH diduga mempengaruhi proses transkripsi gen AMH yang selanjutnya berimplikasi pada pembentukan protein AMH. Apabila terjadi gangguan pada pembentukan protein AMH maka akan berpengaruh terhadap kadar protein tersebut di dalam darah. BAHAN DAN CARA KERJA: Sampel penelitian ini berjumlah 114 pasien yang terdiri dari 60 pasien SOPK dan 54 pasien non SOPK (Kontrol). Kadar AMH dan Jumlah folikel antral didapatkan dari data rekam medik pasien Klinik IVF Yasmin, RSCM Kencana Jakarta. Analisis molekuler dan genotyping dilakukan dengan teknik PCR dan sekuensing kemudian dilanjutkan dengan analisis bioinformatika. HASIL : Dari penelitian ini ditemukan 60 titik varian mutasi promoter gen AMH. Jenis varian mutasi terbesar yang ditemukan adalah -674 G/A (100 %), -245 C/CT (88,2 %), dan -444 A/G (17,9 %) dari seluruh sampel. Berdasarkan hasil uji Wilcoxon Signed Ranks, pada kelompok SOPK ditemukan jumlah mutasi yang terjadi berpengaruh secara bermakna terhadap kadar AMH dan jumlah folikel antral (p<0,05). Pada kelompok kontrol ditemukan bahwa jumlah mutasi tidak berpengaruh secara bermakna terhadap kadar AMH (p>0,05), tetapi berpengaruh secara bermakna terhadap jumlah folikel antral (p<0.05). Ditemukan 60 titik varian pada promoter gen AMH. Jumlah mutasi pada promoter gen AMH berpengaruh terhadap kadar AMH dan jumlah folikel antral pada SOPK. Mutasi pada titik -674 G/A merupakan titik mutasi baru yang belum pernah dilaporkan oleh NCBI, ditemukan pada seluruh subyek penelitian baik kelompok SOPK maupun non SOPK.

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INTRODUCTION : Anti-Mullerian Hormone (AMH) is a member of the Transforming Growth Factor-β group which plays an important role in the regulation of the female reproductive folliculogenesis. A 2-3 fold increase in AMH levels was found in patients with PCOS (Polycystic Ovary Syndrome) compared to women with normal ovulation. This study detected sequence variants in the AMH gene promoter region. The AMH gene promoter variation is thought to affect AMH gene transcription process implicated in the formation of proteins. In the event of disruption in the formation of these AMH proteins, the levels of these proteins in the blood will be affected. The purpose of this study was to detect variants of AMH gene promoter sequences.

MATERIALS AND METHODS: The sample size was 114 patients consisting of 60 PCOS patients and 54 non-PCOS patients as control. The AMH levels and anthral follicle number obtained from the patients? medical records of the Yasmin IVF Clinic, RSCM Kencana Hospital, Jakarta. Molecular analysis and genotyping were performed by PCR and sequencing was followed by bioinformatics analysis.

RESULTS: There were 60 point mutations in the AMH gene promoter variants. The highest variant types of mutations found was -674 G/A (100%), followed by -245 C/CT (88.2%), and -444 A/G (17.9%) in the entire sample. Based on the results of the Wilcoxon Signed Rank test, the number of mutations in the PCOS group were significant to effect the serum AMH level and the anthral follicle number (p<0.05). In the control group, the number of mutations had no significant effect on the levels of AMH (p>0.05), but significantly affected the number of anthral follicles (P<0.05). There were 60 point variances in the AMH gene promoter. The number of mutations in the gene promoter affected serum AMH levels and the number of anthral follicles in PCOS. A new point mutation was found in all subjects at position -674 G/A, which have not been reported by the NCBI."

"Pendahuluan: Sindrom Ovarium Polikistik SOPK merupakan gangguan endokrin tersering penyebab infertifilitas pada wanita usia reproduktif. Wanita-wanita dengan SOPK diketahui memiliki tingkat apoptosis yang rendah dibandingkan dengan wanita tanpa SOPK dan memiliki kadar Anti-Muellerian Hormone AMH yang lebih tinggi dibandingkan dengan wanita tanpa SOPK. Akan tetapi, belum ada penelitian yang menghubungkan kadar AMH dengan apoptosis yaitu adanya atresia folikel pada ovarium wanita penderita SOPKTujuan: Untuk mengetahui hubungan antara kadar AMH yang tinggi dengan tingkat apoptosis sel granulosa yang terjadi pada pasien SOPK sehingga dapat diketahui salah satu patogenesis kelainan folikulogenesis pada pasien SOPKMetodologi: Studi cross sectional dengan mengambil sampel sel granulosa wanita SOPK dan tanpa SOPK atau kelainan ovarium yang mengikuti program Fertiisasi In Vitro FIV di Yasmin dan SMART-IVF, klinik dr. Sander B Jakarta. Jumlah sampel yaitu 40 sampel yang terdiri dari 20 wanita dengan SOPK dan 20 wanita tanpa SOPK. Tingkat apoptosis dievaluasi dengan mengukur ekspresi mRNA dari gen pengkode protein keluarga apoptotic Bcl2 Bax dan Bcl2 menggunakan metode kuantitatif absolut qPCR. Pengukuran kadar AMH di serum dilakukan dengan metode ELISA.Hasil: Terdapat perbedaan bermakna secara statistik antara kadar AMH wanita SOPK dan kontrol pABSTRACT Background Polycystic Ovary Syndrome PCOS is a common endocrine abnormality in causing infertility in reproductive aged women. Women with PCOS were reported have lower apoptosis rate compared to women without PCOS and have higher level Anti Muellerian Hormone compared to women without PCOS. However, there are no reported studies which directly study to know correlation between AMH level in serum and apoptosis result in follicle atresia in ovarium of PCOS patients.Objective To analysis correlation between serum AMH level and apoptosis in granulosa cell in PCOS pasien that may underlie the folliculogenesis abnormality in PCOS.Methods Cross sectional study of sample from granulose cells women with PCOS and without PCOS or with ovarian abnormalities that following Fertility In Vitro FIV program in Yasmine and SMART ndash IVF, dr. Sander B clinic, Jakarta. Sample number were 40 consisting 20 women with PCOS and 20 women without PCOS. Apoptosis level were evaluated with measuring mRNA expression from gene that of coding apoptotic Bcl2 family Bax and Bcl2 using quantitave absolute method qPCR. AMH level in serum were measured using ELISA method.Results There was a statistical significance difference AMH level between PCOS group and control group p"

"Pengantar: Dalam siklus teknologi reproduksi berbantu TRB , sebanyak 30 oosit ditemukan dalam keadaan immatur, oosit immatur ini akan yang memiliki kapasitas maturasi dan fertilisasi yang rendah, dan jarang sampai ketahap embrio transfer, namun faktor-faktor yang menyebabkan terjadinya hal in belum diketahui secara luas. Tujuan: Untuk melihat hubungan antara maturitas oosit dengan kadar hCG serum 12 jam pasca penuntikan dan ekspresi mRNA LHR sel granulosa pada siklus TRB. Untuk menilai apakah kadar hCG serum dan ekspresi LHR ini bisa memprediksi laju maturasi oosit pada siklus TRB. Material dan Metode: total 30 sampel normoresponder yang mengikuti TRB dengan protocol antagonis dianalisa secra prospektif. Dua belas jam setelah penyuntikan hCG, kadar hCG serum diukur dan petik oosit melalui USG transvaginal dilakukan 35-36 jam kemudian. Sel granulosa oosit diperoleh saat denudasi oosit untuk proses intracytoplasmic sperm injection ICSI dan sel granulosa ini kemudian diproses RNA prufikasi, reverse transcription dan quantitative real-time polymerase chain reaction PCR . Oosit yang diperoleh saat itu langsung dinilai maturasinya. Test korelasi Pearson dilakukan untuk menilai korelasi laju maturasi oosit dengan kadar hCG dan ekspresi mRNA LHR. Analisa Receiver Operating Characteristic ROC dilakukan untuk menentukan nilai cut-off. Hasil: Kadar hCG seum memiliki korelasi positif dengan maturitas oosit r 0.467, p

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Introduction: During stimulated in vitro fertilization IVF cycle, up to 30 of the recovered oocytes are immatur ones which have lower maturation capacity, poor fertilization capacity and seldom yield transferable embryos however, the precise influencing factors are largely unknown.

Aim: To investigate the association of oocyte maturation with serum hCG levels measured 12 hours after trigger and LHr mRNA expression of granulosa cell in IVF cycles. To find out whether this serum hCG levels and expression of mRNA LHr granulosa cell can predict oocyte maturation rate in IVF cycles.

Material and Method A total of 30 normoresponder IVF cycles stimulated by antagonist protocol were analyzed prospectively. Twelve hours after triggering by exogenous hCG, level of hCG serum was measured and an ultrasound guided retrieval of oocytes was performed 35 36 hours later. Granulosa cells were obtained during oocyte denudation for intracytoplasmic sperm injection ICSI procedures and subjected to total RNA purification, reverse transcription and quantitative real time polymerase chain reaction PCR. Oocytes were stripped immediately after retrieval and maturation was assessed at this time. Pearson 39 s correlation test performed to analyze the correlation of oocyte maturation rate with serum hCG level and expression mRNA LHR. Receiver operating characteristic ROC analysis was performed to determine cut off value.

Result: Serum hCG have positive correlation with oocyte maturation r 0.467, p"

"ABSTRAK Latar belakang: Penilaian kualitas oosit merupakan bagian terpenting dan tersulit dalam fertilisasi in vitro (FIV). Metode yang saat ini tersedia tidak dapat menilai jumlah dan maturasi oosit secara optimal pada prosedur petik oosit pada FIV. Anti Mullerian Hormone (AMH) dalam cairan folikel dihasilkan langsung oleh sel granulosa, yang diharapkan dapat menjadi parameter alternatif untuk meramalkan kuantitas dan kualitas oosit pada program FIV.Tujuan: Mengembangkan metoda nir-invasif pada program FIV dengan mengetahui nilai prognostik kadar AMH serum dan cairan folikel sebagai parameter alternatif untuk meramalkan kualitas oosit pada program FIV.Metode: Penelitian ini merupakan uji prognostik dengan desain potong lintang selama periode April 2012 hingga Juni 2013 di Poliklinik Yasmin, RSCM, Jakarta. Subjek penelitian merupakan pasien infertilitas yang menjalani program IVF. Pengukuran kadar AMH dan estradiol serum dilakukan pada awal siklus, sementara kadar AMH cairan folikel, jumlah oosit total, jumlah oosit matur dan morfologi oosit dinilai pada saat hari petik ovum. Kualitas oosit dinilai dari skor morfologi oosit modifikasi Xia. Analisis data menggunakan korelasi Pearson dan analisis regresi linier untuk mencari nilai prognostik kadar AMH serum dan cairan folikel sebagai peramal kualitas oosit.Hasil: Didapatkan 102 pasien FIV dengan protokol stimulasi pendek, yang diperiksa kadar AMH serum, kadar AMH cairan folikel, kadar estradiol serum kemudian dinilai jumlah oosit total, jumlah oosit matur dan skor morfologi.Berdasarkan hasil analisis multivariat didapatkan bahwa usia dan AMH serum dapat digunakan dalam meramalkan kualitas oosit (r = - 0,038; r = 0, 183; p < 0,05).Kesimpulan: Kadar AMH serum dan cairan folikel berkorelasi dengan kuantitas oosit. Kadar AMH serum dapat digunakan sebagai parameter untuk meramalkan kualitas oosit, namun tidak untuk AMH cairan folikel

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ABSTRAK Background: Assessment of oocyte quality is an important but difficult component in in vitro fertilization (IVF). Current methods available cannot predict ooctye number and maturation in oocyte pick-up optimally during IVF.Anti Mullerian Hormone (AMH) in follicular fluid is produced directly by granulosa cells, and it could be an alternative parameter to predict oocyte quantity and quality in IVF.Objective: To develop nir-invasif method in IVF program with knowing prognostic value of serum and follicular fluid AMH as alternative parameters to predict oocyte quality.Method: This is a prognostic study with cross sectional design, during a period of April 2012 until June 2013 in Yasmin Clinic, RSCM, Jakarta. The subjects of this study were infertile couples who underwent IVF. The measurement of serum AMH level and serum estradiol serum was done in the beginning of IVF cycles, while follicular fluid AMH, number of oocyte retrieved, number of mature oocyte, and oocyte morphology were measured at the day of ovum pick-up.Oocyte quality using morphological score modified from Xia criteria as parameter. Statistical analysis was done using Pearson correlation and linear regression analysis to measure predictive value of AMH as oocyte qualitypredictors.Result: We obtained 102 short protocol IVF patients. Serum AMH level, AMH level in follicular fluid, serum estradiol, oocyte count, number of mature ooctye, and morphological score were assessed. Based on multivariate analysis, we found that age and serum AMH level can be used to predict oocyte quality (r = - 0,038; r = 0, 183; p < 0,05).Conclusion: There were correlation between serum and follicular fluid AMH with oocyte quantity. AMH level in serum, but not follicular fluid, can be used as a parameter to predict oocyte quality."