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Abstrak :
ABSTRACT
Although icing treatment has been well accepted as aftercare in sports fields, the detailed mechanisms of the treatment is not fully understood. In this study, we investigated the effect of icing treatment on the recovery process of rat plantaris muscles with artificially induced muscle damage. Sixty male Wistar rats (8-weeks-old) were randomly assigned to three groups; control (CTL), bupivacaine-injected (BPVC), and icing treatment after BPVC (ICE). Icing treatment was applied for 20 min immediately after BPVC, and the treatment was used once per day for 3 days. The plantaris muscles were removed at 3, 7, 15, and 28 days after the muscle damage, then immunohistochemical and real time RT-PCR analysis were performed. In histochemical analysis, although significant changes were found in the relative muscle weight, cross-sectional area of muscle fiber, percentage of muscle fiber with central nuclei, and expressed immature myosin heavy chain isoforms after muscle damage, as compared to the CTL group, no differences were found between BPVC and ICE groups. In mRNA expression analysis, the ICE group had a significantly lower value of MyoD than the BPVC group at 3 days after the damage. Expression of IL-6 mRNA, which relates to muscle inflammation, indicated significantly higher value in BPVC, but not in ICE, than CTL groups at 7days after the damage. Furthermore, BKB2 receptor, which relates to acute muscle soreness, indicated a significantly higher expression in BPVC than ICE groups at 3 days after the damage. These results suggest that icing treatment is effective to suppress muscle inflammation and soreness at an early stage of recovery from damage, but not effective for muscle regeneration at a later stage.

Abstrak :
Latar Belakang: Sindrom antifosfolipid (antiphospholipid syndrome = APS) merupakan penyakit autoimun dengan gejala trombosis vena atau arteri, kematian janin berulang, dan peningkatan kadar antibodi antifosfolipid yang persisten. Sindrom antifosfolipid merupakan faktor risiko didapat yang paling sering dihubungkan dengan trombosis. Sampai saat ini efek antibodi anti-?2GP1 pada sistem koagulasi, antikoagulan alamiah dan sistem fibrinolisis masih belum jelas. Tujuan: Menganalisis efek imunoglobulin (Ig)G dan IgM anti-beta-2 glikoprotein-1(?2GP1) terhadap ekspresi messenger RNA (mRNA) tissue factor (TF), mRNA trombomodulin (TM), dan mRNA plasminogen activator inhibitor-1(PAI-1) pada endotel. Metode: Studi eksperimental dengan memajankan antibodi anti-?2GP1 pada human umbilical vein endothelial cells (HUVEC). Penelitian dilakukan di Rumah Sakit Ciptomangunkusumo/ Fakultas Kedokteran Universitas Indonesia. Sampel adalah IgG anti-?2GP1 dan IgM anti-?2GP1 dipurifikasi dari 6 pasien sindrom antifosfolipid. Kontrol adalah IgG dan IgM yang dipurifikasi dari orang sehat. HUVEC dipajan dengan IgG anti-?2GP1, IgM anti-?2GP1, IgG orang sehat, IgM orang sehat selama 4 jam. Pengukuran ekspresi relatif mRNA TF, mRNA TM, dan mRNA PAI-1 dilakukan sebelum dan sesudah pemajanan dengan metode real time reverse transcription polymerase chain reaction. Hasil: Ekspresi relatif mRNA TF, mRNA TM, dan mRNA PAI-1 pada HUVEC yang dipajan dengan IgG anti-?2GP1 adalah (3,14 ± 0,93)-, (0,31 ± 0,13)-, (5,33 ± 2,75)-kali dibandingkan pada HUVEC yang dipajan dengan IgG orang sehat. Ekspresi relatif mRNA TF, mRNA TM, dan mRNA PAI-1 pada HUVEC yang dipajan IgM anti-?2GP1 adalah (4,33 ± 1,98)-, (0,33 ± 0,22)-, (5,47 ± 2.64)-kali dibandingkan pada HUVEC yang dipajan IgM orang sehat. Hasil analisis statistik, sebelum dan sesudah pemajanan HUVEC dengan IgG anti-?2GP1, memperlihatkan perbedaan bermakna ekspresi relatif mRNA TF (1,09 ± 0,76 berbanding 3,14 ± 0,93, p = 0,003), mRNA TM (0,91 ± 0,11 berbanding 0,31 ± 0,13, p = 0,001), dan mRNA PAI-1 (0,93 ± 0,13 berbanding 5,33 ± 2,75, p = 0,013). Hasil analisis statistik, sebelum dan sesudah pemajanan HUVEC dengan IgM anti-?2GP1 memperlihatkan perbedaan bermakna ekspresi relatif mRNA TF (1,03 ± 0,11 berbanding 4,33 ± 1,98, p = 0,008), mRNA TM (0,93 ± 0,08 berbanding 0,33 ± 0,22, p = 0,003), dan mRNA PAI-1 (1,02 ± 0,10 berbanding 5,47 ± 2,64, p = 0,01). Kesimpulan: Pada penelitian ini terbukti bahwa IgG anti-?2GP1 dan IgM anti- ?2GP1 mempunyai efek protrombotik pada sel endotel dengan meningkatkan mRNA TF dan mRNA PAI-1, serta menurunkan mRNA trombomodulin. Hasil penelitian ini menyimpulkan bahwa mekanisme trombosis pada APS dapat terjadi melalui peningkatan aktivasi koagulasi, penurunan aktivitas fibrinolisis dan penurunan aktivitas antikoagulan. ......Background: Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by venous or arterial thrombosis, recurrent pregnancy morbidity and the presence of persistent antiphospholipid antibodies. The antiphospholipid syndrome is the most common acquired risk factor of thrombosis. Until now, the effect of anti-?2GP1 antibodies on coagulation system, natural anticoagulant and fibrinolytic`system has not been completely understood. Objectives: To analyse the effects of IgG and IgM anti-beta-2 glycoprotein-1 (anti-?2GP1) on the expression of tissue factor (TF), thrombomodulin (TM), and plasminogen activator inhibitor-1(PAI-1) of endothelial cells in the messenger RNA level. Methods: Experimental study in human umbilical vein endothelial cells (HUVEC) was done at Cipto Mangunkusumo Hospital/ Faculty of Medicine, Universitas Indonesia. Samples are purified immunoglobulin(Ig)G anti-?2GP1 and IgM anti-?2GP1 from six APS patients serum. For controls, purified IgG and IgM from normal human serum (IgG-NHS and IgM-NHS) were used. HUVEC were treated with purified IgG anti-?2GP1, IgM anti-?2GP1, IgG-NHS, IgM-NHS for four hours of incubation. We measured TF, TM, and PAI-1 of HUVEC in mRNA relative expression levels (before and after treatment) by real time reverse transcription polymerase chain reaction. Results: The mean value of TF, TM, and PAI-1 mRNA levels in HUVEC after treated with IgG anti-?2GP1 compared to Ig-NHS were (3.14 ± 0.93)-, (0.31 ± 0.13)-, (5.33 ± 2.75)-fold respectively. On the other hand, after treated with IgM anti-?2GP1 compared to IgM-NHS, mRNA levels of TF, TM, and PAI-1 were (4.33 ± 1.98)-, (0.33 ± 0.22)-, (5.47 ± 2.64)-fold respectively. Before and after treatment with IgG anti-?2GP1, this study showed significant differences of TF mRNA levels (1.09 ± 0.76 versus 3.14 ± 0.93, p = 0.003), TM mRNA levels (0.91 ± 0.11 versus 0.31 ± 0.13, p = 0.001), and PAI-1 mRNA levels (0.93 ± 0.13 versus 5.33 ± 2.75, p = 0.013). Before and after treatment with IgM anti-?2GP1, this study showed significant differences of TF mRNA levels (1.03 ± 0.11 versus 4.33 ± 1.98, p = 0.008), TM mRNA levels (0.93 ± 0.08 versus 0.33 ± 0.22, p = 0.003), and PAI-1 mRNA levels (1.02 ± 0.10 versus 5.47 ± 2.64, p = 0.01). Conclusion: This study has proven that IgG anti-?2GP1 and IgM anti-?2GP1 increase TF and PAI-1 mRNA levels in endothelial cells. However, IgG anti-?2GP1 and IgM anti-?2GP1 decrease TM mRNA levels in endothelial cells. It has shown that the mechanism of thrombosis in APS occurs through coagulation activation, reduction of fibrinolysis activity, and reduction of anticoagulant activity

Abstrak :
Sindrom ovarium polikistik (SOPK) diketahui terkait dengan obesitas melalui: resistensi leptin. Salah satu penyebab resistensi leptin adalah defisiensi reseptor leptin (LEPR) dibubarkan. Tujuan dari penelitian ini adalah untuk menentukan ekspresi mRNA dari LEPR. gen dilarutkan pada subjek obesitas dan non-obesitas dengan PCOS dan non-SOPK serta mengetahui korelasi antara ekspresi gen tersebut dengan obesitas dan PCOS. Kecepatan Ekspresi mRNA gen LEPR dalam sampel darah diukur menggunakan metode waktu nyata PCR. Penelitian dilakukan pada 96 subjek dengan empat kelompok sampel, yaitu: PCOS non-obesitas, PCOS bebas obesitas, PCOS obesitas, dan PCOS obesitas. Hasil pengukuran menunjukkan ekspresi mRNA rata-rata dari gen LEPR terlarut di masing-masing kelompok 2,10 x 10-4 ng/μL ± 1,88 x 10-4; 1,27 x 10-4 ng/μL ± 1,31 x 10-4; 1,99x 10-4 ng/μL ± 2,35 x 10-4; dan 1,44 x 10-4 ± 2,21 x 10-4 ng/μL. LEPR. ekspresi gen mRNA terlarut dalam semua kelompok obesitas diketahui lebih rendah jika dibandingkan dengan kelompok obesitas (P < 0,05) dan tidak ada perbedaan yang signifikan antara kelompok umum tanpa PCOS dan PCOS (1,69 x 10-4 ± 1,65 x 10-4; 1,71 x 10-4 ± 2,27 x 10- 4, P > 0,05). Studi ini menemukan bahwa penurunan ekspresi mRNA dari gen LEPR yang larut berhubungan dengan obesitas dan tidak berhubungan dengan PCOS.
Polycystic ovary syndrome (SOPK) is known to be associated with obesity through: leptin resistance. One of the causes of leptin resistance is dissolved leptin receptor (LEPR) deficiency. The aim of this study was to determine the mRNA expression of LEPR. gene was dissolved in obese and non-obese subjects with PCOS and non-PCOS as well as determine the correlation between the expression of these genes with obesity and PCOS. Expression velocity of LEPR gene mRNA in blood samples was measured using real-time PCR method. The study was conducted on 96 subjects with four sample groups, namely: non-obese PCOS, obesity-free PCOS, obese PCOS, and obese PCOS. The measurement results showed the average mRNA expression of the soluble LEPR gene in each group was 2.10 x 10-4 ng/μL ± 1.88 x 10-4; 1.27 x 10-4 ng/μL ± 1.31 x 10-4; 1.99x 10-4 ng/μL ± 2.35 x 10-4; and 1.44 x 10-4 ± 2.21 x 10-4 ng/μL. LEPR. soluble mRNA gene expression in all obesity groups was found to be lower when compared to obese group (P < 0.05) and there was no significant difference between the general group without PCOS and PCOS (1.69 x 10-4 ± 1.65 x 10-4; 1.71 x 10-4 ± 2.27 x 10-4, P > 0.05). This study found that decreased mRNA expression of the soluble LEPR gene was associated with obesity and not associated with PCOS.

Abstrak :
LATAR BELAKANG: Endometriosis merupakan penyakit ginekologis kronis yang ditandai dengan adanya jaringan mirip endometrium diluar rongga uterus. Endometriosis merupakan penyebab infertilitas dan nyeri pelvik paling umum dan memengaruhi 1 dari 10 wanita usia reproduktif. Progesteron memiliki peran yang penting dalam uterus yaitu mengontrol proliferasi dan diferensiasi. Disregulasi progesteron dapat menyebabkan gangguan fungsi uterus. Resistensi progesteron banyak ditemukan pada endometriosis dan dikaitkan dengan kadar PGR yang rendah. PGR memiliki dua isoform yaitu PGR-A dan PGR-B. Hingga saat ini masih banyak pendapat yang berbeda mengenai ekspresi isoform PGR-A dan B pada endometriosis. Hipermetilasi pada daerah promotor suatu gen diyakini sebagai salah satu penyebab gene silencing yang berakibat rendahnya kadar gen tersebut. Penelitian ini bertujuan untuk mengetahui pengaruh faktor epigenetik pada reseptor hormon progesteron pada jaringan endometriosis. METODE: Penelitian ini merupakan studi kasus kontrol yang membandingkan 20 wanita penderita endometriosis dan 20 wanita bukan penderita endometriosis. Analisis metilasi DNA dilakukan dengan metode MSP dan ekspresi mRNA dengan metode qPCR. Analisis statistik yang dilakukan adalah uji t tidak berpasangan, uji Mann Whitney, uji korelasi Pearson, uji korelasi Spearman?s Rho, Uji Annova One Way dan Uji Kruskal-wallis dengan kemaknaan p<0.05. HASIL: Hasil penelitian menunjukkan bahwa tingkat metilasi pada wanita dengan endometriosis (98,72%) secara signifikan lebih tinggi dibandingkan dengan kontrol (3,74%) dengan p=0,000. Selain itu, terjadi penurunan ekspresi mRNA isoform PGR-A dan PGR-B (2,35 kali dan 6,37 kali). Terdapat korelasi antara tingkat metilasi dengan ekspresi mRNA isoform PGR-B (p=0,000; r=-0,736), namun tidak terdapat korelasi dengan ekspresi mRNA isoform PGR-A (p>0,05). KESIMPULAN: Daerah promotor gen PGR mengalami hipermetilasi pada endometriosis dibandingkan dengan kontrol dan berkaitan dengan penurunan kadar PGR-B pada endometriosis.

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Background: Endometriosis is a chronic gynecological disorder, defined as the presence of endometrium-like tissue outside of the uterine cavity. Endometriosis is the most common causes of infertility and pelvic pain and affects 1 of 10 women in the reproductive-age group. Progesterone plays an important role in uterine. It controls endometrial proliferation and differentiation. Dysregulation of progesterone signaling leads to impaired for uterine function. Progesterone resistance has been found in endometriosis and associated with the low levels of PGR. PGR has two isoforms, PGR-A and PGR-B. Since promoter hypermethylation is associated with gene silencing, we try to determine the methylation status of PGR promoter region in endometriosis tissue using methylation spesific PCR and its association with the expression of PGR-A and PGR-B isoforms using real-time PCR. Methods: this research is a case-control study, comparing 20 women with endometriosis and 20 women without endometriosis. Methylation status was analyzed with methylaion spesific PCR and the expression of mRNA was analyzed with real-time PCR. Statistical analyses were t-independent test, Mann Whitney test, Pearson test, Spearman?s rho test, Annova One Way test and Kruskal-Wallis test, a two-tailed p value less than 0,05 was considered significant. Result: We found that methylation status in women with endometriosis (98,72%) was significantly higher than women without endometriosis (3,74%), statistically significant associations with the disease (p=0,000). Beside, mRNA expression of PGR-A and PGR-B isoform was down regulated (2,35 fold and 6,37 fold). Correlation between methylation status and PGR-B expression was significant (p=0,000;r=-0,736), but not PGR-A (p>0,05). Conclusion: Promoter regions of PGR is hypermethylated in endometriosis as compared with control. This findings suggest that the promoter hypermethylation of PGR may contribute to the pathogenesis of this disease.

Abstrak :
Latar belakang: Telah dilaporkan bahwa terdapat perubahan pada ekspresi dari ribuan gen di jaringan endometrium endometriosis, termasuk diantaranya adalah gen FN1 dan RAC1. Perubahan ekspresi gen tersebut dapat disebabkan oleh mekanisme epigenetik seperti perubahan tingkat metilasi DNA pada gen. Tujuan: Mengetahui tingkat metilasi DNA pada gen FN1 dan RAC1 serta ekspresi mRNAnya pada jaringan endometrium subjek endometriosis dan nir-endometriosis. Metode: Penelitian ini merupakan cross sectional dengan jumlah sampel sebanyak 40 dari jaringan endometrium subjek endometriosis dan subjek nir-endometriosis. Sampel diambil dengan teknik mikrokuretase di RSUPN Ciptomangunkusumo dan RS Fatmawati Jakarta. Pada jaringan kemudian dilakukan isolasi DNA dan RNA. Pada isolat DNA dilakukan konversi bisulfit, MSP, elektroforesis dan analisis intensitas pita menggunakan software ImageJ untuk mendapatkan data persentase tingkat metilasi DNA. Pada isolat RNA dilakukan qRT-PCR untuk mendapatkan ekspresi relatif mRNA gen FN1 dan RAC1. Hasil: Analisis persentase tingkat metilasi DNA promotor menunjukkan terdapat perbedaan bermakna (p=0,022) pada gen FN1 pada pasien endometriosis (37,95%) dibandingkan nir-endometriosis (59,22 %), sedangkan pada gen RAC1 tidak terdapat perbedaan bermakna (p=0,63) dengan tingkat metilasi subjek endometriosis (28.45%) dan subjek nir-endometriosis (26.11%). Penelitian ini juga melaporkan terjadinya peningkatan ekspresi relatif mRNA gen FN1 dan RAC1 dibandingkan dengan subjek nir-endometriosis, namun secara statistik tidak terdapat perbedaan bermakna (p>0,05). Tidak terdapat korelasi bermakna antara tingkat metilasi gen FN1 dan RAC1 dengan ekspresi mRNAnya. Kesimpulan: Terjadi penurunan tingkat metilasi yang bermakna pada gen FN1 di jaringan endometrium endometriosis, namun tidak berkorelasi dengan peningkatan mRNA nya. Tidak terdapat perbedaan bermakna tingkat metilasi dan ekspresi mRNA pada gen RAC1 di jaringan endometrium subjek endometriosis dibandingkan dengan nir endometriosis.

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It has been reported that there was a changes in the expression of thousands of genes in endometrial endometriosis tissues, including the FN1 and RAC1 genes. Changes in gene expression can be caused by epigenetic mechanisms such as DNA methylation in genes. Objective: To determine the level of DNA methylation in FN1 and RAC1 genes and their mRNA expression in endometrial tissue of endometriosis and non-ndometriosis. Method: This study was designed as cross sectional with a total sample of 40 of endometrial tissues in the subject of endometriosis and non-endometriosis. Samples were taken by microcuretase at Ciptomangunkusumo and Fatmawati Hospital, Jakarta. DNA and RNA was isolated. DNA isolates were converted by bisulfite procedure, MSP conversion, electrophoresis, analyzed intensity of the band which appeared on gel electrophoresis using ImageJ software to obtain the percentage data of DNA methylation level. In RNA isolates, it was analyzed using qRT-PCR methode to obtain the relative mRNA expression level. Results: Analysis of percentage of DNA methylation level showed significant differences (p=0.022) in the FN1 gene (37.95%) compared to non-endometriosis (59.22%), whereas in the RAC1 gene there was no significant difference (p=0,63) with methylation level of endometriosis subjects (28.45%) and non-endometriosis subjects (26.11%). For relative mRNA expression of FN1 and RAC1 genes showed no significant differences (p> 0.05). For correlation in endometrial endometriosis showed no significant between the rate of methylation of the FN1 and RAC1 genes with their mRNA expression. Conclusion: There was a significant decrease in DNA methylation level of FN1 gene in endometrial endometriosis tissues, but it did not correlate with the increasing in its mRNA expression. There was no significant difference in DNA methylation level and mRNA expression of RAC1 gene in endometrial tissues of endometriosis subjects compared to non-endometriosis.

Abstrak :
Tujuan: Menganalisis ekspresi gen manganese superoxide dismutase (MnSOD) pada jaringan jantung, otak dan darah tikus yang diinduksi hipoksia sistemik. Desain: penelitian eksperimental in vivo dengan menggunakan hewan coba. Metode: Sampe! penelitizm ini adalah 25 ekor tikus jantan strain Sprague Dawley (Rarms novergicus L), yang dibagi menjadi 5 kelompok: kelompok I tikus tanpa perlakuan hipoksia sebagai kontrol, kelompok II, III, IV dan V adalah kelompok tikus dengan perlakuan hipoksia 10% O2 selama 1, 7, 14 dan 21 hari. Setelah perlakuan tikus dimaiikan, kemudian darah, otak dan jantung tikus diambil untuk diperiksa tingkat ekspresi mRNA dengan menggunakan real time RT PCR dengan pewamaan SYBR green, serta diukur aktivitas spesifik MnSOD dengan menggunakan kit RanSOD® dengan ditambahkan NaCN untuk menghambat aktivitas CuZn SOD. Hasil: Pada hipoksia awa] (1 hari) ekspresi relatif mRNA MnSOD dan aktivitas spesifik MnSOD menunjukkan penurunan di darah dan jantung, sedangkan pada otak tidak te1jadi penurunan. Hal ini menunjukkan bahwa dalam keadaan hipoksia sistemik perlindungan antioksidan pada otak terjadi lebih awal dibandingkan jantung dan darah. Pada hipoksia awal di jantung dan darah, mulai terjadi peningkatan ROS sehingga aktivitas spesink MnSOD menurun, namun belum dapat menstimulasi peningkatan eksprsi mRNA-nya_ Pada hipoksia I-I4 hari baik ekspresi mRNA maupun aktivitas spesiiik MnSOD pada ketiga jaringan tersebut mengalami peningkatan sejalan dengan lamanya hipoksia. Pada hipoksia lanjut (21 hari) terjadi korelasi negatif antara ekspresi relatif mRNA dngan aktivitas spesiiik MnSOD di jantung dan darah. Hal ini mnmgkin disebabkan karena produksi ROS yang sangat masif, sehingga ekspresi MRNA terus ditingkatkan namun stres oksidatif belum dapat diatasi, sedangkan pada otak fenomena tersebut tidak terjadi. Hal ini diduga karena peningkatan ROS pada hipoksia lanjut masih dapat diatasi dengan aktivitas enzim MnSOD yang tersedia tanpa harus meningkatkan ekspresi mRNA-nya. Hasil ini menunjukkan bahwa otak cenderung lebih dilindungi dalam keadaan hipoksia sistemik dibandingkan janrung dan darah. Hasil analisis uji korelasi Pearson menunjukkan bahwa perubahan ekspresi relatif MRNA dan aktivitas spesifik MnSOD pada induksi hipoksia sistemik pada darah sejalan dengan perubahannya pada jantung dan otak. Kesimpulan: Setiap jaringan mempunyai pola ekspresi gen MnSOD dan aktivitas MnSOD yang berbeda-beda pada kondisi hipoksia. Terdapat perbedaan regulasi ekspresi gen MnSOD antara hipoksia sistemik awal dan lanjut. Pengukuran ekspresi MnSOD (mRNA dan aktivitas spesifik) pada darah dapat sekaligus menggambarkan ekspresi tersebut pada jantung dan otak.

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Background: The aim of this study is to determine the gene expression of manganese supenoxide dismutase (MnSOD) in rat?s heart, brain and blood induced by systemic hypoxia. Design: This study is an in vivo experimental study. Method: This study was conducted on 25 male Sprague Dawley rats (Rattus no1°e:~_gicn.s~ L) which were divided into 5 groups and subjected to systemic hypoxia by placing them in hypoxic chamber supplied by 10% O3 for O, l, 7. I4, 2.1 days. respectively. Rats were sacrified after treatment, and the blood. heart and brain were used for measurement of relative mRNA level ofMnSOD with real time RT PCR and measurement of spesitic activity of MnSOD enzyme using RanSOD® kit. Result: Determination of gene expression of MnSOD (relative mRNA expression and specific activity) in rat blood and heart cells under early hypoxic induction (1 day) resulted in the lower levels compared to the level in control group. After l day of hypoxic induction the gene expression level was then increased and again decreased under very late hypoxic condition (21 days) compared to the control. This suggests that the blood and heart cells at early hypoxia have not enough time to provide more MnSOD enzyme through gene expression to eliminate the sudden accumulation of ROS. In contrast to the results in heart and blood cells. the gene expression of MnSOD in brain cells were demonstrated to be increased since early systemic hypoxia (day I) up to day l4_ and tends to decrease under late hypoxic condition (day 21) although the level still slightly higher compared to the level in control group. Under late hypoxic condition (21 days). the capacity of1VlnSOD to eliminate the accumulated ROS has been saturated as found in brain cells, or even reduced to the lower level than in normal condition as found in blood and heart cells. This study could demonstrate that brain cells have different pattern of gene expression of MnSOD compared to blood and heart cells during several time points of hypoxic induction, particularly at early stage. It should also be considered that the levels of gene expression of MnSOD in each tissue were distinct although measured under the same condition. Analysis of Pearson correlation test shows that pattern of gene expression ot`MnSOD in blood cells is appropriate with the pattern in heart and brain cells under hypoxic condition. Conclusion: Every tissue has the different pattern of gene expression of MnSOD (relative mRNA expression and specific activity) under hypoxic condition There is different regulation of MnSOD gene expression at early and late hypoxia Analysis gene expression of MnSOD in blood cells could represent the analysis of gene expression of MnSOD in heart and brain cells under hypoxia condition.

Abstrak :

Sindrom ovarium polikistik (SOPK) merupakan kelainan reproduksi yang ditandai dengan anovulasi, menstruasi tidak teratur, dan hirsutisme yang seringkali menyebabkan infertilitas. Meski etiologinya belum sepenuhnya dipahami, namun telah diketahui bahwa sebagian besar wanita dengan SOPK mengalami obesitas dengan prevalensi mencapai 40—80%. Obesitas merupakan kelebihan akumulasi lemak tubuh yang dicirikan dengan hipertrofi. Salah satu gen yang diduga terkait dengan obesitas adalah gen fat mass and obesity associated (FTO). Gen FTO menyebabkan peningkatkan nilai BMI. Penelitian ini bertujuan untuk mengetahui perbedaan ekspresi mRNA gen FTO dan korelasinya dengan BMI pada wanita SOPK dan normal dengan obesitas dan non-obesitas. Jaringan darah digunakan sebagai sumber mRNA yang diambil pada 30 wanita non obesitas, 30 wanita normal obesitas, 30 wanita SOPK non-obesitas, dan 30 wanita SOPK obesitas. Kuantitas ekspresi mRNA gen FTO ditentukan dengan menggunakan quantitative real-time PCR. Hasil penelitian menunjukkan terdapat perbedaan ekspresi mRNA gen FTO pada kelompok wanita SOPK dan normal dengan obesitas, serta tidak terdapat korelasi antara ekspresi mRNA gen FTO dengan BMI. Gen FTO merupakan gen yang bertanggung jawab terhadap obesitas akan tetapi tidak memiliki keterkaitan dengan sindrom ovarium polikistik, serta tidak memiliki korelasi dengan BMI.

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Polycystic ovary syndrome (PCOS) is a reproductive disorder characterized by anovulation, irregular menstruation, and hirsutism which often causes infertility. Although the etiology is not fully understood, it is well known that most women with PCOS are obese with a prevalence of 40-80%. Obesity is an excess of body fat accumulation which is characterized by hypertrophy. Fat mass and obesity associated gene (FTO) is known to correlate with obesity. The study found that FTO gene causes an increase in the BMI. This reserach aim to determine the differences in FTO mRNA gene expression and its correlation with BMI in normal and PCOS woman with obesity and lean. This study used blood tissue as a source of mRNA taken in 30 normal lean woman, 30 normal obese women, 30 PCOS lean women, and 30 PCOS obese women. The quantity of FTO mRNA gene expression was determined using quantitative real-time PCR. The result shows that there is differences in FTO mRNA gene expression in PCOS and normal woman with obesity dan non-obesity. FTO mRNA expression in PCOS and normal obesity woman is higher than those in the PCOS and normal lean women, and there is no correlation between FTO gene mRNA expression and BMI. Thus, the FTO gene is a gene responsible for obesity but has no association with polycystic ovary syndrome, and does not have a correlation with BMI. 

Abstrak :
Pengembangan vaksin berbasis mRNA merupakan teknologi yang berkembang pesat untuk mengobati penyakit menular serta menawarkan beberapa keunggulan dibandingkan vaksin konvensional. Namun, stabilitas vaksin mRNA menjadi tantangan utama dalam pengembangannya sehingga digunakan nanopartikel lipid (LNP) sebagai sistem penghantarannya karena memiliki kemampuan untuk memperbaiki stabilitasnya. Komponen penyusun LNP yang digunakan pada penelitian ini adalah CTAB, DSPC, kolesterol, dan DMG-PEG 2000 dengan memvariasikan konsentrasi CTAB dan kolesterol menjadi tiga formula untuk mendapatkan hasil formulasi terbaik. Konsentrasi yang divariasikan yaitu CTAB:DSPC:kolesterol:DMG-PEG 2000 secara berturut-turut adalah 13,5:20:65:1,5% (F1); 18,5:20:60:1,5% (F2); dan 23,5:20:55:1,5 % (F3). Formulasi LNP dibuat menggunakan metode t-junction mixing dengan kecepatan laju alir total sebesar 700 mL/jam dan volume akhir LNP sebanyak 20 mL. Pengaruh variasi rasio konsentrasi tersebut terhadap nilai ukuran partikel, indeks polidispersitas, dan potensial zeta diukur menggunakan Particle Size Analyzer serta RT-qPCR untuk mengidentifikasi adanya RNA yang terjerap dalam sampel LNP-mRNA. Hasil terbaik didapatkan dari formulasi LNP-mRNA kedua dengan rasio variasi lipid CTAB:DSPC:kolesterol:DMG-PEG 2000 sebesar 18,5:20:60:1,5 % yang menghasilkan rata-rata ± standar deviasi ukuran partikel sebesar 257,54 ± 9,11 nm; indeks polidispersitas sebesar 0,245 ± 0,01; dan potensial zeta sebesar +2,1 ± 0,16 mV. Setelah dilakukannya analisis kualitatif dengan metode RT-qPCR, ditemukan adanya mRNA dalam sampel LNP-mRNA. Penelitian ini memberikan wawasan baru dalam formulasi LNP-mRNA dengan menggunakan konsentrasi surfaktan kationik seperti CTAB sebesar 18,5% dan lipid helper seperti kolesterol sebesar 60%. ......The development of mRNA-based vaccines is rapidly evolving as a technology to treat infectious diseases, offering several advantages over conventional vaccines. However, the stability of mRNA vaccines remains a major challenge in their development. To address this, lipid nanoparticles (LNP) are used as delivery systems because of their ability to improve stability. The components of LNP used in this study include CTAB, DSPC, cholesterol, and DMG-PEG 2000, with varying concentrations of CTAB and cholesterol across three formulations to achieve the best results. The varying concentrations were CTAB:DSPC:cholesterol:DMG-PEG 2000 at ratios of 13.5:20:65:1.5% (F1); 18.5:20:60:1.5% (F2); and 23.5:20:55:1.5% (F3). The LNP formulations were prepared using the T-junction mixing method with a total flow rate of 700 mL/hour and a final LNP volume of 20 mL. The impact of these concentration ratios on its particle size, polydispersity index, and zeta potential was measured using a Particle Size Analyzer, and RT-qPCR was used to identify the presence of RNA encapsulated in the LNP-mRNA samples. The best results were obtained from the second LNP-mRNA formulation with a lipid ratio of CTAB:DSPC:cholesterol:DMG-PEG 2000 at 18.5:20:60:1.5%, producing an average ± standard deviation particle size of 257.54 ± 9.11 nm, a polydispersity index of 0.245 ± 0.01, and a zeta potential of +2.1 ± 0.16 mV. Qualitative analysis using the RT-qPCR method confirmed the presence of mRNA in the LNP-mRNA samples. This study provides new insights into LNP-mRNA formulation using cationic surfactant concentrations like CTAB at 18.5% and helper lipids like cholesterol at 60%.

Abstrak :
ABSTRAK
Latar belakang. Proses inflamasi kronik dan persisten mempengaruhi tingginya rekurensi dan survival endometriosis pasca pembedahan. Hal ini menjadi permasalahan endometriosis, sehingga perlu pengembangan terapi target salah satunya yaitu asam galat. Asam galat terbukti efektif sebagai antikanker, anti tumor, anti inflamasi dan antibakterial pada beberapa cell line, namun efektifitasnya pada sel endometriosis harus dibuktikan. Tujuan. membuktikan efek asam galat dan senyawa turunannya terhadap regulasi inflamasi pada kultur primer endometriosis ditinjau dari ekspresi mRNA NF-kB, serta sekresi TNF-? dan IL-6. Metode. Sel endometriosis berasal dari jaringan endometriosis pasien yang menjalani laparaskopi, diisolasi secara enzimatis dan dikultur primer. Sel kultur diberi perlakuan asam galat, heptil dan oktil galat dengan dosis 25,6 g/mL, 51,2 g/mL dan 102,4 g/mL selama 48 jam, kemudian diinduksi dengan LPS 500 ng/mL selama 24 jam. Regulasi inflamasi dinilai dari ekspresi mRNA NF-kB dengan qRT-PCR, kadar sekresi TNF-? dan IL-6 dengan ELISA, serta inhibisi viabilitas sel dengan MTS Assay. Hasil. Setelah data dirasiokan dengan kontrol, ketiga zat signifikan menghambat viabilitas sel endometriosis p value 0,000 dengan inhibisi tertinggi pada dosis 102,4 g/mL. Terjadi penurunan ekspresi relatif NF-kB yang dirasiokan dengan kontrol dan IL-6 meskipun secara statistik tidak bermakna. Konsentrasi TNF? tidak berbeda secara bermakna p value 0,340 . Kesimpulan. Asam galat dan senyawa turunannya berpengaruh terhadap inhibisi viabilitas sel, penurunan ekspresi relatif NF-kB dan IL-6, namun tidak bermakna terhadap penekanan sitokin TNF-?. Perlu dilakukan studi lanjut untuk menilai efektifitas asam galat sebagai kandidat obat antiinflamasi pada endometriosis ditinjau aspek lain.

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ABSTRACT
Background. Endometriosis is a benign gynecological disorder characterized by the growth of the lining of the endometrium like tissue outside the uterus. The cause of the growth of endometriosis is not known well, chronic and persistent inflammatory process is suspected to be one of the pathogenesis that contributes to the high recurrence and survival endometriosis. One of the potential therapeutic agents is a gallic acid which proved effective in earlier studies as an anti cancer, anti tumor, anti inflammatory and antibacterial in several cell line. The Effectiveness of gallic acid to the endometriosis cell is a preliminary study and have not found evidence of publication yet. Object. Proving the effect of gallic acid and its derivatives on the inflammatory regulation of endometriosis primary culture study on mRNA expression of NF kB, TNF , and IL 6 secretion. Method. Endometriosis cells from Indonesian endometriosis patients tissues who had undergone laparoscopy surgery were isolated by the enzymatic reaction and primary cultured. Cultured cells treated by gallic acid and alkyl ester synthetic derivatives of the gallic acids heptyl gallate and octyl gallate each with the dosage of 25,6 g mL, 51,2 g mL, and 102.4 g mL for 48 hours and then induced by LPS 500 ng mL for 24 hours. Parameter research was assessed by qRT PCR for mRNA expression of NF kB, ELISA for the quantification of TNF and interleukin 6, and MTS assay was used to observe endometriosis cell viability. Results. After the data was rationalized with the control, three substances showed significant inhibition of endometriosis cell viability. The highest inhibition for all treatment was at doses 102,4 g mL. Overall there was an inhibition of relative expression of mRNA NF kB were rationalized to controls and suppression of IL 6 in octyl gallate groups. The concentration of TNF among the groups did not differ significantly p value 0.340 . Conclusion. Gallic acid and its derivatives have significantly inhibition effect toward cell viability, mRNA expression of NF kB, and IL 6 but have not significantly effect toward cytokine TNF . Further studies need to be conducted to assess the effectiveness of gallic acid as an anti inflammatory drug candidate toward to any pathway.

Abstrak :
Latar belakang: Sindrom ovarium polikistik (SOPK) merupakan salah satu kelainan endokrin paling umum pada 7-15% wanita usia reproduksi yang menyebabkan infertilitas anovulatori. SOPK sering ditemukan pada wanita gemuk, sekitar 50-75% tetapi sindrom ini juga dapat ditemukan pada wanita kurus sebesar 5,5%. Penyebab dan patogenesis SOPK sampai sekarang masih diperdebatkan tapi hasil penelitian memperlihatkan hiperandrogen dan resistensi insulin terlibat dalam perkembangan perjalanan penyakit dan fenotip SOPK. Efek androgen dimediasi oleh reseptor androgen (AR) sedangkan efek insulin dimediasi oleh reseptor insulin (INSR). Ekspresi dan aksi dari kedua reseptor ini terutama pada sel granulosa ovarium dapat dipengaruhi oleh mekanisme epigenetik yang diduga terlibat dalam perkembangan penyakit SOPK ini. Tujuan: Mengetahui tingkat metilasi DNA pada gen reseptor androgen (AR) dan reseptor insulin (INSR) serta ekspresi mRNAnya pada sel granulosa subjek SOPK dan nir-SPOK. Metode: Penelitian ini merupakan penelitian deskriptif analitik dengan rancangan cross sectional. Sampel berupa sel granulosa dari folikel ovarium didapatkan dari wanita yang melakukan ovum pick up (OPU) di klinik Yasmin RSUPN Ciptomangunkusumo yang kemudian dilakukan isolasi DNA dan RNA. Pada isolat DNA dilakukan konversi bisulfit, Methyl Specifik PCR (MSP), elektroforesis dan analisis ketebalan pita dengan perangkat lunak ImageJ untuk mendapatkan data tingkat metilasi DNA. Pada isolat RNA dilakukan qPCR untuk mendapatkan ekspresi relatif mRNA gen AR dan INSR. Hasil: Analisis data dari 21 subjek SOPK dan 20 subjek nir SOPK menunjukkan terdapat perbedaan bermakna (p=0,00) tingkat metilasi DNA gen AR pada pasien SOPK (45.24 %±16,84) dibandingkan nir-SOPK (84,96±15,45). Analisis gen INSR, pada subjek SOPK 100% tidak terjadi metilasi pada promotor gen INSR tetapi pada subjek nir-SOPK 1 dari 21 wanita mengalami metilasi parsial ((37,82%±8,25). Dari penelitian juga didapatkan terjadinya peningkatan ekspresi relatif mRNA AR sebesar 2,459 kali dan 1,791 kali pada mRNA INSR. Tidak terdapat korelasi antara tingkat metilasi gen AR dan INSR dengan ekspresi mRNAnya. Kesimpulan: Penurunan tingkat metilasi DNA (hipometilasi) pada gen AR dan INSR dapat meningkatkan tingkat ekspresi mRNA AR dan INSR, yang kemudian berkontribusi terhadap kejadian hiperandrogen dan resistensi insulin pada fenotip subjek SOPK. ......Background: Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in 7-15% of women of reproductive age who cause anovulatory infertility. PCOS is often found in obese women, around 50-75% but this syndrome can also be found in thin women at 5.5%. The causes and pathogenesis of PCOS is still debated but the results of the study show hyperandrogen and insulin resistance involved in the development of the disease course and the PCOS phenotype. The androgen effect is mediated by the androgen receptor (AR) while the effect of insulin is mediated by the insulin receptor (INSR). The expression and action of these two receptors, especially in ovarian granulosa cells can be influenced by epigenetic mechanisms that are thought to be involved in the development of PCOS. Objective: To determine the level of DNA methylation in the androgen receptor gene (AR) and insulin receptor (INSR) and its mRNA expression in the SOPK and nir-SPOK granulosa cells. Method: This study is a descriptive analytic study with a cross sectional design. Samples in the form of granulosa cells from ovarian follicles were obtained from women who performed ovum pick up (OPU) at the Yasmin clinic Ciptomangunkusumo Hospital which was then isolated from DNA and RNA. DNA isolates were carried out bisulfite conversion, Methyl Specific PCR (MSP), electrophoresis and tape thickness analysis with ImageJ software to obtain DNA methylation level data. QPCR was performed on RNA isolates to obtain the relative expression of the AR and INSR mRNA genes. Results: Analysis of data from 21 SOPK subjects and 20 non-PCOS subjects showed significant differences (p = 0.00) of AR gene DNA methylation rates in PCOS patients (45.24% ± 16.84) compared to non-PCOS (84.96 ± 15 , 45). INSR gene analysis, in the subject of 100% PCOS there was no methylation of the INSR gene promoter but in non-PCOS subjects 1 out of 21 women had partial methylation ((37.82% ± 8.25). AR is 3.459 times and 2.791 times in INSR mRNA. There is no correlation between the rate of methylation of the AR and INSR genes with their mRNA expression. Conclusion: Decreasing levels of DNA methylation (hypomethylation) in AR and INSR genes can increase the level of expression of mRNA AR and INSR, which then contributes to the incidence of hyperandrogen and insulin resistance in the phenotype of the PCOS subject.