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"Ikan Tiger shovelnose catfish Pseudoplatystoma fasciatum (Linnaeus, 1766) merupakan ikan hias introduksi yang memiliki pertumbuhan cepat. Pertumbuhan berperan penting pada perkembangan ikan dan dipengaruhi kinerja hormon pertumbuhan (GH). Hormon pertumbuhan pada ikan jumlahnya terbatas, sehingga perlu dilakukan perbanyakan melalui isolasi gen GH, agar dapat diaplikasikan dalam peningkatan produktivitas ikan. Penelitian ini bertujuan untuk mengisolasi dan menganalisis ekspresi mRNA gen GH pada ikan Tiger shovelnose catfish. Isolasi GH dilakukan dari jaringan kelenjar hipofisis pada ikan berukuran 602 g dan 43 cm. Tahapan isolasi diawali ekstraksi RNA, sintesis cDNA, dan Reverse Transcription Polymerase Chain Reaction (RT-PCR) menggunakan primer GH degenerate dari data 7 spesies catfish di gene bank, serta gen b-actin sebagai kontrol internal. Gen GH selanjutnya di-cloning dan sequencing. Ekspresi gen GH pada tahap perkembangan awal diamati sejak stadia embrio, larva (3, 10, dan 15 dph, day post hatched) dan juvenil (20, 45, dan 60 dph), kemudian dianalisis secara semi kuantitatif. Data ekspresi gen dianalisis menggunakan uji ANOVA satu arah dan dilanjutkan uji Tukey. Isolasi mRNA gen GH telah berhasil dilakukan secara parsial, dengan panjang sekuens 234 bp dan b-actin berukuran 300 bp. Gen GH ikan Tiger shovelnose catfish secara homology dekat dengan ikan patin (Pangasianodon hypophthalmus) dan ikan lele (Clarias batracus) dengan nilai sama yaitu 90,60%. Gen GH mulai terekspresi sejak dari stadia embrio. Ekspresi gen GH menurun pada dari stadia larva ke juvenil, karena merupakan tahap metamofosis. Stadia juvenil merupakan level ekspresi tertinggi (P<0,05), karena organ ikan sudah lebih lengkap dan ekspresinya akan terus meningkat seiring pertambahan usia.

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An ornamental fish, the Tiger Shovelnose Catfish Pseudoplatystoma fasciatum (Linnaeus, 1766) grows quickly. Growth hormone affects the performance of growth and development in this species. Because the amount of growth hormone in this fish is limited, it is necessary to isolate the GH gene to increase fish productivity. Accordingly, the aim of study is to isolated and to determine mRNA level of GH gene from each stage. The mRNA GH gene was isolated from 602 g of fish pituitary tissue. Followed by the b-actin gene used as an internal control in Reverse Transcription Polymerase Chain Reaction RT-PCR utilizing degenerate GH primers from 7 catfish species in the gene bank. The GH gene was then sequenced. GH gene expression was measured semi-quantitatively in embryonic, larval (3, 10, and 15 dph), and juvenile (20, 45, and 60 dph) stages, respectively. Gene expression of each stage were analyzed by one-way ANOVA and was followed by Tukey's test. The partial isolation of GH gene mRNA has been successfully carried out, with a sequence length of 234 bp and gene of b-actin at 300 bp. The GH gene of Tiger shovelnose catfish was homology close to catfish (Pangasianodon hypophthalmus) and catfish (Clarias batracus) with the same value of 90.60%. GH gene expression decreased from larval to juvenile stage, because it was a metamorphosis stage. Juvenile stage is the highest expression level (P<0.05), because fish organs are more complete and their expression will continue to increase with age."

"Ikan Tiger shovelnose catfish Pseudoplatystoma fasciatum (Linnaeus, 1766) merupakan ikan hias introduksi yang memiliki pertumbuhan cepat. Pertumbuhan berperan penting pada perkembangan ikan dan dipengaruhi kinerja hormon pertumbuhan (GH). Hormon pertumbuhan pada ikan jumlahnya terbatas, sehingga perlu dilakukan perbanyakan melalui isolasi gen GH, agar dapat diaplikasikan dalam peningkatan produktivitas ikan. Penelitian ini bertujuan untuk mengisolasi dan menganalisis ekspresi mRNA gen GH pada ikan Tiger shovelnose catfish. Isolasi GH dilakukan dari jaringan kelenjar hipofisis pada ikan berukuran 602 g dan 43 cm. Tahapan isolasi diawali ekstraksi RNA, sintesis cDNA, dan Reverse Transcription Polymerase Chain Reaction (RT-PCR) menggunakan primer GH degenerate dari data 7 spesies catfish di gene bank, serta gen -actin sebagai kontrol internal. Gen GH selanjutnya di-cloning dan sequencing. Ekspresi gen GH pada tahap perkembangan awal diamati sejak stadia embrio, larva (3, 10, dan 15 dph, day post hatched) dan juvenil (20, 45, dan 60 dph), kemudian dianalisis secara semi kuantitatif. Data ekspresi gen dianalisis menggunakan uji ANOVA satu arah dan dilanjutkan uji Tukey. Isolasi mRNA gen GH telah berhasil dilakukan secara parsial, dengan panjang sekuens 234 bp dan -actin berukuran 300 bp. Gen GH ikan Tiger shovelnose catfish secara homology dekat dengan ikan patin (Pangasianodon hypophthalmus) dan ikan lele (Clarias batracus) dengan nilai sama yaitu 90,60%. Gen GH mulai terekspresi sejak dari stadia embrio. Ekspresi gen GH menurun pada dari stadia larva ke juvenil, karena merupakan tahap metamofosis. Stadia juvenil merupakan level ekspresi tertinggi (P<0,05), karena organ ikan sudah lebih lengkap dan ekspresinya akan terus meningkat seiring pertambahan usia.

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An ornamental fish, the Tiger Shovelnose Catfish Pseudoplatystoma fasciatum (Linnaeus, 1766) grows quickly. Growth hormone affects the performance of growth and development in this species. Because the amount of growth hormone in this fish is limited, it is necessary to isolate the GH gene to increase fish productivity. Accordingly, the aim of study is to isolated and to determine mRNA level of GH gene from each stage. The mRNA GH gene was isolated from 602 g of fish pituitary tissue. Followed by the -actin gene used as an internal control in Reverse Transcription Polymerase Chain Reaction RT-PCR utilizing degenerate GH primers from 7 catfish species in the gene bank. The GH gene was then sequenced. GH gene expression was measured semi-quantitatively in embryonic, larval (3, 10, and 15 dph), and juvenile (20, 45, and 60 dph) stages, respectively. Gene expression of each stage were analyzed by one-way ANOVA and was followed by Tukey's test. The partial isolation of GH gene mRNA has been successfully carried out, with a sequence length of 234 bp and gene of -actin at 300 bp. The GH gene of Tiger shovelnose catfish was homology close to catfish (Pangasianodon hypophthalmus) and catfish (Clarias batracus) with the same value of 90.60%. GH gene expression decreased from larval to juvenile stage, because it was a metamorphosis stage. Juvenile stage is the highest expression level (P<0.05), because fish organs are more complete and their expression will continue to increase with age."

"Gen tilapia Growth Hormone (tiGH) merupakan gen pengkode hormon pertumbuhan dari ikan nila yang berperan untuk meningkatkan pertumbuhan. Penelitian bertujuan melakukan kloning dan ekspresi gen tiGH untuk memproduksi protein rekombinan hormon pertumbuhan ikan nila (Oreochromis niloticus). Penelitian meliputi tahapan isolasi gen tiGH dari pMBA_tiGH, ligasi ke dalam pETBlue-2, serta transformasi vektor rekombinan ke dalam sel inang dengan menggunakan elektroporasi. Hasil penelitian menunjukkan bahwa vektor rekombinan dapat ditransformasi ke dalam sel inang E. coli BL2 dengan efisiensi transformasi 1,12x103 cfu/μg. Ekspresi gen tiGH dilakukan menggunakan induksi IPTG 0,4 mM dan dipurifikasi menjadi protein rekombinan growth hormone dengan berat molekul sebesar 22 kDa.

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Tilapia growth hormone gene (tiGH) is a gene encoding growth hormone from the tilapia whose folr is to increase the growth. The research objective is to do cloning and expression tiGH gene to produce growth hormone recombinant proteins of tilapia (Oreochromis niloticus). Stages of research include isolation tiGH gene from pMBA_tiGH, ligation into pETBlue-2, and the transformation recombinant vector into host cells by using electroporation.

The result showed that recombinant vectors have been successfully transformed into the host cell E.coli BL21with transformation efficiency reached 1.12 x103 cfu/μg. Expression tiGH gene performed using 0.4 mM IPTG induction and purified recombinant protein growth hormone with a molecular weight of 22 kDa."

"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."

"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."

"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 sertamengetahui 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 dengankelompok 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 asdetermine 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."

"Kanker ovarium epitelial (EOC) merupakan kanker ginekologi paling mematikan yang salah satunya disebabkan karena progresivitasnya yang cepat. Berdasarkan tipe molekuler dan progresivitasnya, EOC dapat dibagi menjadi tipe high-grade dan low-grade. Pada tahapan karsinogenesis EOC seringkali melibatkan reaksi  inflamasi. Siklooksigenase‑2 diketahui berkontribusi pada progresivitas tumor dan dapat teraktivasi oleh jalur pensinyalan NF-κB . Penelitian ini bertujuan untuk menganalisis korelasi varian genetik RELA rs11820062 dan PTGS2 rs20417 beserta ekspresi mRNA RELA dan PTGS2 di EOC pada pasien di Rumah Sakit Cipto Mangunkusumo, Indonesia. Adanya variasi RELA rs11820062 dan PTGS2 rs20417 diduga akan memengaruhi ekspresi mRNA RELA dan PTGS2. Penelitian ini menggunakan 65 sampel darah wanita normal dan total 80 biopsi ovarium dengan rincian: 15 kista ovarium sebagai kalibrator ekspresi, 36 EOC low-grade, dan 29 EOC high-grade. Distribusi genotipe dan alel dianalisis menggunakan ARMS PCR dan ekspresi mRNA menggunakan teknik qPCR. Tidak ditemukan perbedaan distribusi genotipe dan alel kedua varian antara kelompok normal dan EOC. Ekspresi mRNA RELA dan PTGS2 signifikan lebih tinggi pada kelompok EOC dibandingkan kelompok kista. Genotipe TT dari rs11820062 berkorelasi dengan ekspresi mRNA RELA yang lebih rendah pada EOC high-grade dan low-grade. Sedangkan genotipe PTGS2 rs20417 tidak berkorelasi dengan tingginya ekspresi mRNA PTGS2 pada EOC. Ekspresi RELA dan PTGS2 diketahui saling berkorelasi satu sama lain.

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Epithelial ovarian cancer (EOC) is the highest mortality type of gynecology cancer due to its progressivity. EOC can be divided based on its molecular type and progressivity into low-grade and high-grade EOC. Every step of EOC’s carcinogenesis is known to involve an inflammation reaction. Cyclooxygenase‑2 is one of the inflammatory factors contributing to tumor progression and can be activated by the NF-κB signaling pathway. Hence, we examined their distribution and correlation to RELA and PTGS2 mRNA expression in EOCs patients from Cipto Mangunkusumo Hospital, Indonesia. A total of 65 healthy subjects and 80 ovarian biopsies (15 ovarian cysts as expression calibrators, 36 low-grade EOC, and 29 high-grade EOC) were used in this study. The distribution of genotypes and alleles was analyzed using ARMS PCR and mRNA expression using the qPCR technique. There was no significant difference between genotype and allele distributions for both variants. Both genes’ mRNA relative expression was significantly higher in EOC than in ovarian cyst. RELA rs11820062 TT genotype is correlated with lower RELA mRNA relative expression in EOC. In contrast, PTGS2 rs20417 didn’t correlate with the considerably higher PTGS2 mRNA relative expression in EOC tissues. The increase of PTGS2 mRNA relative expression is known to correlate with RELA mRNA relative expression."

"Pendahuluan: Kanker payudara triple-negatif merupakan subset keganasan yang menantang terkait dengan prognosis buruk akibat rendahnya ekspresi HER2 dan reseptor hormon, yang mengakibatkan kurangnya modalitas terapi yang ditargetkan secara spesifik. Selain itu, bagian dari keganasan ini mempunyai tingkat kekambuhan dini dan metastasis yang tinggi. Manganese superoxide dismutase (MnSOD), suatu enzim yang sangat penting untuk keseimbangan redoks, terlibat dalam tumorigenesis karena peran gandanya. Awalnya, MnSOD bertindak sebagai penekan tumor selama tumorigenesis awal, namun perannya bergeser seiring perkembangan penyakit. Kanker payudara triple-negatif diperkaya dengan subpopulasi sel dengan potensi tumorigenik tinggi dan stres oksidatif yang dikenal sebagai sel punca kanker. Sel-sel ini mengekspresikan faktor transkripsi Oct4 dan Sox2, yang mengatur pembaruan diri dan kepuncaan. Studi ini menyelidiki efek penekanan ekspresi MnSOD melalui KO gen CRISPR/Cas9 pada sel punca kanker BCSC triple-negatif dengan mengukur ekspresi mRNA OCT4 dan SOX2. Metode: Penelitian ini menggunakan kelompok kontrol BCSC tripel-negatif BT-549 tipe wild-type dan dua kelompok BT-549 KO MnSOD yang diberi perlakuan. Untuk setiap kelompok, tiga sampel ulangan digunakan. Ekspresi mRNA OCT4 dan SOX2 di setiap kelompok kontrol dan kelompok perlakuan dideteksi oleh RT-qPCR. Hasil masing-masing dihitung dengan Metode Livak untuk mengekstraksi rasio ekspresi. Uji T dua sampel dilakukan untuk menghitung signifikansinya. Hasil: Temuan ini menunjukkan tren penurunan ekspresi SOX2 dan ekspresi mRNA OCT4 yang tidak konsisten. Kesimpulan: Penekanan MnSOD dapat menjadi target potensial untuk mengubah sifat pluripotensi BCSC triple-negatif dengan memberikan efek tidak langsung pada ekspresi mRNA OCT4 dan SOX2.

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Introduction: Triple-negative breast cancer represents a challenging subset of malignancy associated with poor prognosis due to low expression of HER2 and hormone receptors, resulting in the lack of specific targeted therapeutic modalities. Additionally, this subset of malignancy acquires a high rate of early recurrence and metastasis. Manganese superoxide dismutase (MnSOD), an enzyme paramount for redox balance, is implicated in tumorigenesis by its dual role. Initially, MnSOD acts as a tumor suppressor during early tumorigenesis, but its role shifts as the disease advances. Triple-negative breast cancer is enriched with a subpopulation of cells with high tumorigenic potential and oxidative stress known as cancer stem cells (CSC). These cells expressed Oct4 and Sox2 transcription factors, which regulate self-renewal and stemness. This study investigates the effect of suppressing MnSOD expression through CRISPR/Cas9 gene knockout on the stemness of triple-negative BCSCs by measuring OCT4 and SOX2 mRNA expression. Methods: This study utilized a control group of wild-type BT-549 triple-negative BCSCs and two treated groups of MnSOD-knockout BT549 BCSCs. For each group, three replicate samples were used. The mRNA expression of OCT4 and SOX2 in each control and treated group was detected by RT-qPCR. Their respective results were calculated by the Livak Method to extract the expression ratio. A two-sample T-test was performed to calculate the significance. Results: The findings demonstrate a decreasing trend of SOX2 expression and an inconsistent OCT4 mRNA expression. Conclusion: MnSOD suppression may present as a potential target for altering the pluripotency properties of triple-negative BCSCs by exerting an indirect effect on OCT4 and SOX2 mRNA expression."

"Latar Belakang: Sindroma ovarium polikistik (SOPK) merupakan sebuah penyakti dengan prevalensi yang tinggi dan beban kesehatan yang besar. Hingga saat ini, penyebab SOPK masih belum jelas. Penelitian sebelumnya mengenai ekspresi reseptor vitamin D (VDR) pada pasien SOPK menunjukkan hasil yang menjanjikan namun belum terbukti secara jelas. Oleh sebab itu, diduga bahwa polimorfisme VDR berperan penting dalam kejadian dan beratnya gejala SOPK.Metode: Penelitian potong lintang dilakukan pada pasien SOPK dan wanita usia reproduktif non-SOPK sebagai kontrol pada November 2019 hingga 2021. Pasien hamil, menyusui, memiliki riwayat gangguan hormon adrenal, tiroid, maupun prolaktin, atau mengonsumsi obat hormonal dalam 6 bulan terakhir dieksklusi dari penelitian. Subjek penelitian direkrut secara konsekutif. Ekspresi VDR dinilai dari ekspresi mRNA VDR yang dinilai dengan pemeriksaan PCR. Polimorfisme VDR dinilai pada tiga titik regio penyandi gen, yakni rs7975232, rs11574113, dan rs11574114.Hasil: Sebanyak 80 pasien SOPK dan 80 pasien kontrol diikutsertakan dalam penelitian. genotip A/A pada regio rs7975232 dan genotip C/C pada regio rs11574113 lebih banyak didapatkan pada pasien dengan SOPK. Di sisi lain, genotip A/C pada regio rs7975232 dan genotip C/G pada regio rs11574114 lebih banyak didapatkan pada kelompok kontrol (p < 0,05). Tidak terdapat perbedaan ekspresi VDR pada pasien dengan polimorfisme yang berbeda (p > 0,05).Kesimpulan: Didapatkan polimorfisme gen penyandi VDR yang berbeda antara pasien SOPK dan non-SOPK. Tidak didapatkan perbedaan ekspresi VDR yang bermakna antara pasien SOPK dan non-SOPK.

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Background: Polycystic ovary syndrome (SOPK) is a disease with a high prevalence and a large health burden. Until now, the cause of SOPK is still unclear. Previous studies on vitamin D receptor (VDR) expression in PCOS patients have shown promising results but have not been clearly proven. Therefore, it is suspected that the VDR polymorphism plays an important role in the incidence and severity of PCOS symptoms.

Methods: A cross-sectional study was conducted on PCOS patients and non-SOPK women of reproductive age as controls from November 2019 to 2021. Patients who were pregnant, breastfeeding, had a history of adrenal, thyroid, or prolactin hormone disorders, or had taken hormonal drugs in the last 6 months were excluded from the study. study. Research subjects were recruited consecutively. VDR expression was assessed from VDR mRNA expression assessed by PCR examination. VDR polymorphism was assessed at three points in the gene encoding region, namely rs7975232, rs11574113, and rs11574114.

Results: A total of 80 PCOS patients and 80 control patients were included in the study. A/A genotypes in the rs7975232 region and C/C genotypes in the rs11574113 region were more common in patients with PCOS. On the other hand, the A/C genotype in the rs7975232 region and the C/G genotype in the rs11574114 region were more common in the control group (p < 0.05). There was no difference in VDR expression in patients with different polymorphisms (p > 0.05).

Conclusions: Different polymorphisms of the VDR coding gene were found between PCOS and non-SOPK patients. There was no significant difference in VDR expression between PCOS and non-SOPK patients."

"Plastik merupakan salah satu barang yang memiliki tingkat penggunaan yang tinggi hingga mencapai skala produksi sebesar 300 juta ton pertahun. Limbah plastik di perairan membutuhkan waktu hingga ratusan tahun untuk terdegradasi karena karakteristiknya yang kuat, tahan lama, dan tahan terhadap suhu tinggi. Proses degradasi limbah plastik di perairan disebabkan oleh sinar UV yang menyebabkan terjadinya perubahan bentuk dan ukuran menjadi lebih kecil (mikroplastik) atau tercampur di dalam air (< 5mm). Ukuran yang kecil menyebabkan mikroplastik dapat dengan mudah termakan oleh biota perairan, seperti ikan atau melalui mangsanya yang lebih kecil. Penelitian ini bertujuan untuk menganalisis komposisi dan kelimpahan mikroplastik berdasarkan bentuk, ukuran, dan polimer pada air, sedimen, serta organ dan jaringan ikan kiper Scatophagus argus (Linnaeus, 1766) di muara Sungai Blanakan, Subang, Jawa Barat. Metode penelitian dilakukan dengan pengambilan sampel air sebanyak 50 L lalu disaring menggunakan plankton net hingga mendapatkan 1 L sampel air, pengambilan sedimen dilakukan menggunakan Van Veen grab lalu dimasukkan ke dalam jar HDPE 500 ml, dan pengambilan sampel ikan sebanyak 10 individu dilakukan menggunakan alat tangkap bubu. Sampel air ditambahkan larutan H2O2 30% dan FeSO4 0,05 M masing-masing sebanyak 25 ml. Sampel sedimen dicampurkan larutan NaCl dengan perbandingan sebesar 1 : 2 dan didiamkan. H2O2 30% dan FeSO4 0,05 M ditambahkan hingga sampel terendam. Sampel ikan dibedah untuk diambil bagian insang, saluran pencernaan, dan daging. Setiap bagian dihancurkan menggunakan larutan KOH 10% dengan perbandingan 1 : 10 dan didiamkan. Sampel air, sedimen, dan ikan dipipet sebanyak 40 ml dan disaring menggunakan kertas saring cellulose nitrate dengan bantuan vacuum pump lalu kertas saring ditempatkan di cawan petri. Sampel air, sedimen, dan ikan kiper diamati pada kertas saring menggunakan mikroskop Olympus CX22 dan analisis polimer mikroplastik dilakukan dengan metode Raman Spectroscopy. Analisis statistik dilakukan menggunakan uji one way ANOVA, Kruskal wallis, dan Pearson. Hasil penelitian menunjukkan rerata kelimpahan mikroplastik pada sampel air sebesar 710 partikel/m3, sedimen sebesar 879,63 partikel/kg, dan ikan sebesar 74,77 partikel/individu. Kelimpahan mikroplastik dari yang tertinggi hingga terkecil pada ikan kiper terdapat di daging, saluran pencernaan, dan insang. Komposisi mikroplastik paling dominan berupa bentuk fiber dan ukuran < 300 µm. Polimer yang ditemukan bertipe PET, PP, dan PVC. Uji Kruskal wallis menunjukkan terdapat perbedaan secara nyata kelimpahan mikroplastik antar organ dan jaringan ikan. Uji Pearson menunjukkan adanya korelasi kelimpahan mikroplastik pada air dan sedimen terhadap kelimpahan mikroplastik di ikan.

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Plastic is one of the commodities with a high level of usage, reaching a production scale of 300 million tons per year. Plastic waste in water takes hundreds of years to degrade due to its strong, durable, and heat-resistant characteristics. The degradation process of plastic waste in water is caused by UV radiation, which leads to changes in shape and size, resulting in smaller particles (microplastics) or mixing with water (< 5mm). The small size of microplastics makes them easily ingestible by aquatic organisms, such as fish, or through their prey, which are smaller in size. This research aims to analyze the composition and abundance of microplastics based on their form, size, and polymer in water, sediments, as well as the organs and tissues of the Scatophagus argus(Linnaeus, 1766) fish in the estuary of the Blanakan River, Subang, West Java. The research method involved collecting 50 liters of water samples, which were then filtered using a plankton net to obtain a 1-liter water sample. Sediment samples were collected using a Van Veen grab and placed in a 500 ml HDPE jar, while fish samples were collected using a fish trap, with a total of 10 individuals. The water sample was treated with 25 ml of 30% H2O2 and 0.05 M FeSO4 solutions. The sample was mixed with a NaCl solution in a ratio of 1:2 and left to settle. H2O2 (30%) and FeSO4 (0.05 M) were added until the sample was fully submerged. The samples were dissected to obtain gill, digestive tract, and muscle tissue. Each part was crushed using a 10% KOH solution in a 1:10 ratio and left to settle. Water, sediment, and fish samples were pipetted (40 ml) and filtered using cellulose nitrate filter paper with the assistance of a vacuum pump, and the filter papers were placed in Petri dishes. The water, sediment, and fish samples were observed on the filter paper using an Olympus CX22 microscope, and the analysis of microplastic polymers was conducted using Raman Spectroscopy. Statistical analysis was performed using one-way ANOVA, Kruskal-Wallis, and Pearson tests. The results of the study showed that the average abundance of microplastics in water samples was 710 particles/m3, in sediments it was 879.63 particles/kg, and in fish it was 74.77 particles/individual. The highest abundance of microplastics in the kiper fish was found in the muscle tissue, followed by the digestive tract and gills. The dominant composition of microplastics was in the form of fibers and with a size of < 300 µm. The polymers found were PET, PP, and PVC. The Kruskal-Wallis test indicated a significant difference in microplastic abundance among the organs and tissues of fish. The Pearson test showed a correlation between the abundance of microplastics in water and sediments with the abundance of microplastics in fish."