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"ABSTRAKLatar belakang: SOPK adalah gangguan endokrin yang hingga saat ini etiologinya masih belum jelas. Faktor epigenetik metilasi DNA, akhir-akhir ini mendapatkan perhatian dalam patogenesis SOPK. Gen HSD17B1 disebut sebagai "estrogenik" 17β-HSD karena mengkatalisasi langkah terakhir dalam biosintesis estrogen dengan secara istimewa mengurangi estrone, estrogen yang lemah untuk menghasilkan estrogen 17β-estradiol yang kuat. Kami berspekulasi cacat pada metilasi DNA mendorong deregulasi gen sehingga terjadi penurunan ekspresi mRNA HSD17B1, akhirnya menghasilkan estradiol yang tidak cukup pada pasien SOPK.Metode: Kami mengumpulkan total 60 pasien wanita. MSP untuk analisis metilasi DNA, qPCR untuk analisis ekspresi mRNA.Tujuan: Untuk menganlisis metilasi DNA pada kelompok pasien SOPKdan kelompok wanita sehat, ekspresi mRNA pada kelompok pasien SOPK dan kelompok wanita sehat, tingkat estradiol pada pasien SOPK dan kelompok wanita sehat, korelasi antara metilasi DNA dan ekspresi mRNA pada pasien SOPK, korelasi ekspresi mRNA pada pasien SOPKdan kadar serum estradiol.Hasil: Metilasi gen HSD17B1 pada wanita SOPK adalah 42,64% dan kelompok yang sehat menunjukkan 53,80%, p = 0,160 tidak signifikansi antara kedua kelompok. Nilai ekspresi relatif gen HSD17B1 adalah 0,70 kali lebih rendah dibandingkan dengan kelompok wanita sehat, p = 0,003 signifikansi antara kedua kelompok. Estradiol rata-rata pada kelompok SOPK25,78 pg / ml dan kelompok wanita sehat adalah 36,74 pg / ml. Korelasi tingkat metilasi DNA versus ekspresi mRNA pada pasien SOPK, tidak signifikan p = 0,076. Korelasi antara ekspresi mRNA gen HSD17B1 dan kadar serum estradiol, signifikansi p = 0,020. ;Semakin terjadi penurunan ekspresi mRNA, semakin rendah kadar serum estradiol.

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ABSTRACTBackground: PCOS is the most common endocrine disorder but its etiology remains unclear. Lately, epigenetic factors have gained considerable attention in the pathogenesis of PCOS, DNA methylation.  HSD17B1 is referred to as the "estrogenic" 17β-HSD because it catalyzes the final step in estrogen biosynthesis by preferentially reducing the weak estrogen estrone to yield the potent estrogen 17β-estradiol. We speculated defects in DNA methylation promote the deregulation of genes make decrease mRNA expression HSD17B1, finally produces not enough estradiol in PCOS patients.Methods: We collected a total of 60 female patients. MSP for DNA methylation analysis, qPCR for mRNA expression analysis.Aims: To investigate, DNA methylation in PCOS patients group and healthy women group, mRNA expression in PCOS patients group and healthy women group, estradiol level in PCOS patients and healthy women group, the correlation between DNA methylation and mRNA expression in PCOS patients, correlation mRNA expression in PCOS patients and estradiol serum level.Results: Methylated of HSD17B1 gene in PCOS women was 42.64 % and a healthy group showed 53.80 %, p=0.160 not significances between the two groups.  The relative expression value of the HSD17B1 gene was 0.70 fold lower compare with a healthy women group, p=0.003 significance between the two groups. The average estradiol in the PCOS group 25.78 pg/ml and the healthy women group is 36.74 pg/ml. Correlation of DNA methylation level versus mRNA expression in PCOS patients, not significance p=0.076. Correlation between mRNA expression of the HSD17B1 gene and estradiol serum level, significance p=0.020. (More decrease mRNA expression, more lower estradiol serum level)."

"Endometriosis sering dikaitkan dengan nyeri menstruasi dan nyeri pelvis. Penyakit ini terjadi sekitar 10-15% pada perempuan usia reproduksi . NGF, TRPA1 dan reseptor P2RX3 aktivitas gen terlibat dalam respon nyeri. Penelitian ini bertujuan untuk menganalisis tingkat ekspresi mRNA gen NGF, reseptor TRPA1 dan reseptor P2RX3 yang diduga disebabkan oleh perubahan tingkat metilasi DNA promoter gen tersebut, serta hubungannya pada intensitas nyeri subjek endometriosis.Sampel jaringan endometrium dan susukan endometriosis peritoneum diperoleh dari 20 subjek endometriosis, sementara jaringan endometrium kontrol diperoleh dari 20 subjek nir endometriosis. Metode yang digunakan untuk analisis metilasi DNA yaitu metode MSP dan perangkat lunak Image-J digunakan untuk menganalisis intensitas pita metilasi dari gen NGF, reseptor TRPA1 dan reseptor P2RX3R, selanjutnya digunakan metode qRT-PCR untuk analisis tingkat mRNA gen-gen tersebut. Penilaian intensitas nyeri dilakukan dengan menggunakan kuesioner standar skala penilaian numerik (NRS) melalui wawancara dengan pasien. Pada penelitian ini didapatkan hasil terdapat hubungan yang bermakna antara intensitas nyeri dan kejadian endometriosis (p>0,001). Hasil tersebut dibuktikan dengan terdapat perbedaan yang bermakna tingkat ekspresi relatif mRNA gen NGF, reseptor P2RX3 antara jaringan endometrium endometriosis dibandingkan dengan subjek nir endometriosis masing-masing nilai p= <0,05. Terdapat juga perbedaan yang bermakna tingkat metilasi DNA promotor gen NGF dan reseptor P2RX3 antara jaringan endometrium endometriosis dengan endometrium nir endometriosis (p<0,05). Selanjutnya, terdapat hubungan yang bermakna antara ekspesi mRNA NGF dan reseptor Reseptor P2RX3 dengan intensitas nyeri pada endometriosis (p<0,05), namun tidak terdapat hubungan antara tingkat metilasi DNA dengan ekspresi relatif mRNA gen NGF, reseptor reseptor TRPA1 pada endometriosis begitupun antara metilasi DNA dengan intensitas nyeri pada ketiga gen tersebut (p>0,05). Terjadi perubahan ekspresi relatif mRNA gen NGF, reseptor TRPA1 dan reseptor P2RX3 pada subjek endometriosis yang berhubungan dengan peningkatan intensitas nyeri pada endometriosis, namun mekanisme epigenetik metilasi DNA pada penelitian tersebut tidak berhubungan pada intensitas nyeri endometriosis.......Endometriosis is often associated with both cyclic menstrual pain and pelvic pain. It affects 10% of reproductive age women. NGF, TRPA1 and P2RX3 receptors gene activity are found to be involved in pain response. This study aims to analyze the methylation level of NGF gene, TRPA1 and P2RX3 receptors that might alter the mRNA expression in peritoneal endometriosis and endometrial tissue, as well as its correlation to the pain level in endometriosis patients.20 endometrial tissues and 20 peritoneal endometriosis tissues were obtained from patients, while 20 endometrium tissues as control were obtained from healthy women. First, each participant was given informed consent before the research begin. We used methylation specific PCR (MSP) and Image-J software to analyze the methylation level of NGF gene, TRPA1 and P2RX3 receptors; electrophoresis to analyze the band intensity; qRT-PCR to evaluate the mRNA level in each gene. Finally, we evaluated the pain level using the standardized questionnaire of numeric rating scale (NRS) by doing interviews with patients. In this study, it was found that there is a significant relationship between pain intensity and the incidence of endometriosis (p> 0.001). This results is proven by a significant difference in the mRNA expression level of NGF gene and P2RX3 receptor between endometrial endometriosis and endometrial non-endometriosis tissues, with each p value = <0.05. There is also a significant difference in the DNA methylation level of NGF gene and P2RX3 receptor between endometrial endometriosis and endometrial non-endometriosis tissues (p <0.05). There is a significant relationship between the mRNA expression level of NGF gene, P2RX3 receptor and pain intensity in endometrial endometriosis tissues (p <0.05). However, the results showed that there is no correlation between the DNA methylation level and the mRNA expression of NGF gene, TRPA1 and P2RX3 receptors. There is also no correlation between the DNA methylation level of NGF gene, TRPA1 and P2RX3 receptors and pain intensity in endometriosis tissue. There is an alteration of mRNA expression of NGF gene, TRPA1 and P2RX3 receptors which correlates to pain intensity in endometriosis patients. However, there is no correlation between DNA methylation level and pain intensity in endometriosis patients."

"Latar belakang: Matrix metalloproteinases (MMPs) merupakan protein yang berperan dalam proses inflamasi dan remodeling yang disebabkan oleh infeksi, termasuk tuberkulosis paru (TB), terutama multidrug resistance. Penelitian ini bertujuan untuk mengkorelasikan hubungan antara kadar serum dan polimorfisme MMP-1 dan MMP-9 dengan karakteristik kavitas, seperti jumlah, diameter, ketebalan dinding, dan distribusi fibrosis pada Multidrug-Resistant (MDR) dan Drug-Sensitive (DS) pasien TB.Metode: Penelitian ini menggunakan desain studi potong lintang komparatif. Subyek yang berasal dari pasien rawat jalan RS Abdoel Moelok Lampung Indonesia telah lulus uji etik. Subjek dibagi menjadi dua kelompok, 34 subjek pada kelompok MDR-TB dan 36 subjek pada kelompok DS-TB. Kadar protein serum MMP-1 dan MMP-9 dilakuakn dengan uji ELISA, dan genotipe MMP-1 dan MMP-9 dengan Sequencing metode Sanger. Kemudian kavitas dan fibrosis dievaluasi dengan menggunakan pemeriksaan High-Resolution Computerized Tomography (HRCT) toraks.Hasil: Terdapat perbedaan bermakna jumlah kavitas dengan diameter lebih dari 6, 6 mm, dan tebal kavitas pada pasien TB-MDR dibandingkan dengan pasien TB-DS. Distribusi fibrosis pada segmen paru juga berbeda nyata pada MDR-TB dibandingkan dengan DS-TB. Walaupun kadar MMP-9 pada kelompok MDR-TB lebih tinggi dibandingkan dengan kelompok DS-TB, namun secara statistik tidak terdapat perbedaan yang signifikan dari penelitian yang menunjukkan bahwa terdapat hubungan antara MDR-TB dan DS-TB mengenai jumlah kavitas, diameter kavitas, ketebalan dinding kavitas, serta distribusi fibrosis di segmen paru-paru yang terkena yang dievaluasi dengan HRCT. Penelitian ini mendapatkan frekuensi alel G pada MMP-1 pada populasi Indonesia (Asia) dan adanya hubungan yang signifikan dengan tebal kavitas dengan alel G pada MMP-1 dan alel T pada MMP-9 alelKesimpulan: Tidak terdapat hubungan antara genotipe MMP-1 (-1607G) dan MMP-9 (C1562T) dengan kadar serum MMP-1 dan MMP-9, genotipe MMP 1 pada kedua kelompok penelitian berbeda secara bermakna dan merupakan faktor pencegahan dua kali lipat kejadian MDR-TB. Selain itu, terdapat perbedaan yang substansial dalam ketebalan dinding kavitas antara genotipe G/G MMP-1 1607 T/T MMP-9 pada kedua kelompok penelitian.......Background: Matrix metalloproteinases (MMPs) are proteins that play a role in the inflammatory and remodeling processes caused by infections, including pulmonary tuberculosis (TB), especially multidrug resistance. This study aims to correlate the relationship between serum levels and polymorphism of MMP-1 and MMP-9 with cavity characteristics, such as number, diameter, wall thickness, and distribution of fibrosis in Multidrug-Resistant (MDR)- and Drug-Sensitive (DS)-TB patients.Method: This study used a comparative cross-sectional study design. The subjects came from outpatients at Abdoel Moelok Hospital, Lampung Indonesia had passed the ethical test. Subjects were divided into two groups, 34 subjects in the MDR-TB group and 36 subjects in the DS-TB group. The levels of MMP-1 and MMP-9 were carried out by ELISA test, and the genotipes MMP-1 and MMP-9 were determined using PCR-the Sequencing method. In addition, cavities and fibrosis were measured using thoracic High- Resolution Computerized Tomography (HRCT) imaging.Results: There was a significant difference in the number of cavities with a diameter of more than 6.6 mm, and cavity thickness in MDR-TB patients compared to DS-TB patients. The distribution of fibrosis in the lung segments was also significantly different in MDR-TB compared to DS-TB. Although MMP-9 levels in the MDR-TB group were higher than in the DS-TB group, there was no statistically significant difference from the study, which showed a relationship between MDR-TB and DS-TB regarding the number of cavities, cavity diameter, walls thickness cavity, as well as the distribution of fibrosis in the affected lung segments evaluated by HRCT. This study found the frequency of the G allele in MMP-1 in the Indonesian population (Asia) and a significant relationship with cavity thickness between the G allele in MMP-1 and the T allele in MMP-9.Conclusion: There is no relationship between the MMP-1 (-1607G) and MMP-9 (C1562T) genotypes with serum levels of MMP-1 and MMP-9, the MMP 1 genotype in the two study groups was significantly different and was a factor preventing twice the incidence MDR-TB. In addition, the two study groups showed substantial differences in cavity wall thickness between the G/G MMP-1 1607 T/T MMP-9 genotypes."

"Latar Belakang: Infertilitas dialami oleh sekitar 15% pasangan di seluruh dunia, dengan kontribusi dari pihak laki-laki sekitar 50%. Salah satu penyebab infertilitas pada pria adalah azoospermia non obstruktif idiopatik, yang diduga melibatkan faktor epigenetik. Penelitian ini bertujuan menilai peran epigenetik, khususnya remodeling kromatin dan modifikasi histon, pada proses spermatogenesis pada testis dengan azoospermia non obstruktif.Metode: Sampel BFPE dan TESE diperiksa menggunakan teknik HE lalu dikelompokkan berdasarkan tipe henti maturasi, yaitu SCO, STA, dan SDA. Sampel BFPE dilakukan pemeriksaan immunohistokimia menggunakan antibodi anti-CHD5, anti-H3K9me3, dan anti-H4K12ac. Proses pengolahan gambar immunohistokimia menggunakan ImageJ, IHC Profiler, dan StarDist. Sampel TESE dilakukan pemeriksaan qPCR untuk mengukur tingkat ekspresi gen CHD5 dan PHF7. Selain itu, pada sampel TESE dilakukan pemeriksaan ChIP untuk menilai kadar relatif gen WEE1 dan PRM1 yang berikatan dengan CHD5.Hasil: Ekspresi CHD5 ditemukan pada spermatogonia dan spermatid bulat. Tidak ada perbedaan signifikan intensitas CHD5 pada spermatogonia antara kelompok STA dan SDA. Intensitas H3K9me3 dan H4K12ac pada spermatogonia, spermatosit, dan sel sertoli berdasarkan kelompok henti maturasi berbeda signifikan. Tingkat ekspresi gen CHD5 pada kelompok STA meningkat signifikan 67 kali lipat dibandingkan ekspresinya pada SCO, dan pada kelompok SDA meningkat signifikan 164 kali lipat dibandingkan ekspresi pada SCO. Tingkat ekspresi gen PHF7 pada kelompok STA meningkat signifikan 53 kali lipat dibandingkan ekspresinya pada SCO, dan pada kelompok SDA meningkat signifikan 192 kali lipat dibandingkan ekspresi pada SCO. Kadar DNA segmen promoter gen WEE1 pada ChiP-qPCR menggunakan antibodi anti-CHD5 ditemukan sebesar 1,19% untuk STA dan 1,87% untuk SDA, lebih tinggi dibandingkan kadar pada SCO yaitu 0,36%. Sedangkan kadar DNA segmen promoter gen PRM1 ditemukan sebesar 1,01% untuk STA dan 2,47% untuk SDA, lebih tinggi dibandingkan kadar pada SCO yaitu 0,29%.Kesimpulan: CHD5 berperan pada spermatogenesis manusia, khususnya pada sel spermatogonia dan spermatid bulat. CHD5 terbukti meregulasi gen WEE1 dan PRM1 pada sel spermatogenik. H3K9me3 dan H4K12ac berperan pada kasus henti maturasi, dan berpotensi untuk menjadi marker kasus azoospermia non obstruktif.......Background: Infertility affect about 15% of couples worldwide, with male factors contributing to around 50% of cases. One of the causes of male infertility is idiopathic non-obstructive azoospermia, which is suspected to involve epigenetic factors. This study aims to assess the role of epigenetics, specifically chromatin remodeling and histone modification, in the process of spermatogenesis in testes with non-obstructive azoospermia.Method: The BFPE and TESE samples were examined using HE techniques and subsequently classified based on maturation arrest types, including SCO, STA, and SDA. Immunohistochemical analysis of the BFPE samples was conducted using anti-CHD5, anti-H3K9me3, and anti-H4K12ac antibodies. Image processing for immunohistochemistry was performed using ImageJ, IHC Profiler, and StarDist. The TESE samples underwent qPCR analysis to measure the expression levels of the CHD5 and PHF7 genes. Additionally, ChIP analysis was performed on the TESE samples to assess the relative levels of WEE1 and PRM1 genes bound to CHD5.Result: The expression of CHD5 was found in spermatogonia and round spermatids. There was no significant difference in CHD5 intensity in spermatogonia between the STA and SDA groups. However, the intensities of H3K9me3 and H4K12ac in spermatogonia, spermatocytes, and Sertoli cells varied significantly among the maturation arrest groups. The expression level of the CHD5 gene in the STA group increased significantly by 67-fold compared to its expression in SCO, and in the SDA group, it increased significantly by 164-fold compared to its expression in SCO. The expression level of the PHF7 gene in the STA group increased significantly by 53-fold compared to its expression in SCO, and in the SDA group, it increased significantly by 192-fold compared to its expression in SCO. The DNA segment promoter level of the WEE1 gene in ChiP-qPCR using anti-CHD5 antibody was found to be 1.19% for STA and 1.87% for SDA, higher than the level in SCO, which was 0.36%. Meanwhile, the DNA segment promoter level of the PRM1 gene was found to be 1.01% for STA and 2.47% for SDA, higher than the level in SCO, which was 0.29%.Conclusion: CHD5 plays a role in human spermatogenesis, particularly in spermatogonia and round spermatids. It has been shown to regulate the genes WEE1 and PRM1 in spermatogenic cells. H3K9me3 and H4K12ac are implicated in cases of maturation arrest and have potential as markers for azoospermia non obstructive cases."