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The Expression of Homo Sapiens microRNA-21 (Hsa-miR-21-5p) and mRNA Reversion Inducing Cysteine Rich Protein with Kazal Motifs (RECK) in Plasma of Epithelial Ovarian Cancer Kartika, Aprilia Indra; Chasanah, SN; Fitriawan, AS; Tanjung, DS; Ftria, MS; Pakun, FK; Oktriani, R; Trirahmanto, A; Prajatmo, H; Aryandono, T; Haryana, SM
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 48, No 4 (2016): SUPPLEMENT
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

ABSTRACT Epithelial Ovarian Cancer (EOC) is malignant cancer that caused death for most women in Indonesia. The emergence of EOC showed no specific symptoms in its early stages; that makes the screening mostly occur when patients are in advanced stage. Treatment of EOC at an advanced stage will be more challenging with poor prognosis. Therefore, minimally invasive biomarkers are needed to diagnose at the early stage. microRNA is one of the potential biomarkers which not only expressed inside the cell but also secreted outside the cell with exosome protection. This protection makes microRNA stable. Moreover, several studies have shown the ability to detect microRNA in the blood sample. microRNA-21 (miR-21) is oncomiR which targeted tumor suppressor mRNA RECK based on in silico analysis.The first aim is to determine the expression of miR-21 in plasma samples of EOC patients compared with healthy controls. The second aim is to investigate the expression correlation between miR-21 and RECK mRNA.Blood samples were collected from 30 patients and 30 healthy controls. Plasma was then obtained from centrifugated blood samples. The total RNA was isolated and reverse transcribed to produce cDNAs. cDNAs were then quantified using qPCR using specific primer for miR-21 and RECK mRNA. The expression analysis was done relative expression method by Livak. The expression of miR-21 was calculated using the miR-16 expression as the reference gene. Also, Beta-actin was used as reference gene for RECK mRNA calculation. The correlation between the expression of miR-21 and mRNA RECK was analyzed using the Spearman rho correlation analysis.In this study showed the expression of miR-21 in patients with EOC increased 4.7579-fold compared with healthy controls (p <0.05). On the other hand, the miR-21 target, RECK mRNA, decreased 4,2 times with fold change 0.237665 on plasma EOC patients compared with healthy controls (p <0.05). The statistical calculation of the expression of miR-21 and mRNA RECK was inversely proportional to mRNA RECK with a strong correlationThis study has been able to prove that the expression of miR-21 is up-regulated in EOC patients and confirmed by the down-regulation of RECK expression. The next research challenge is to make anti-miR-21 to suppress the expression of miR-21 in EOC, which can be analyzed the effect of miR-21 in the development of EOC.
The Expression of hsa-miR-155-5p in Plasma Samples Of Breast Cancer Before And After Chemotherapy Fitria, Meutia Srikandi; Haryana, Sofia Mubarika; Anwar, Sumadi Lukman; Aryandono, Teguh; Tanjung, Dewi Sahfitri; Kartika, Aprilia Indra; Oktriani, Risky; Irianianiwati, .; Sari, Dwi Nur Indah
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 48, No 4 (2016): SUPPLEMENT
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Abstract

Breast cancer has emerged as the most common cancer-related mortality among women worldwide. Therefore, early cancer detection using biomarkers such as microRNA is needed. One of microRNAs that has an important role in breast cancer development is miR-155. Hsa-miR-155-5p is an oncomir that is commonly dysregulated in breast cancer. This study aims to determine the expression of hsa-miR-155-5p in breast cancer patient?s plasma before and after chemotherapy. We collected 64 samples from breast cancer patients admitted to Dr. Sardjito Hospital in Yogyakarta. RNA from plasma was extracted using RNA Isolation Kit miRCURY-Biofluid. cDNA synthesis was performed using cDNA Synthesis kit II and quantification of miR-155-5p using ExiLent SYBR Green master mix (Exiqon). qRT-PCR results were then analyzed with Livak's method and compared (before and after chemotherapy) with t-test. Expression of miR-155-5p in the breast cancer patients? plasma after chemotherapy was significantly increased (10.59 times) when compared to before chemotherapy (p = 0.001). We concluded that there was upregulated expression of miR-155-5p after chemotherapy than before chemotherapy. There has not been a known, relevant pathway between hsa-miR-155-5p and chemotherapy regimens nor resistance to chemotherapy.?Keywords: Breast cancer, plasma, hsa-miR-155-5p, oncomiR, chemotherapy.
MENINGKATKAN NILAI AFEKTIF DAN HASIL BELAJAR MATERI NATURAL PHENOMENA MELALUI MODEL SCIENTIFIC INQUIRY Kartika, Aprilia Indra; Pukan, Krispinus Kedati; Mustikaningtyas, Dewi
Journal of Biology Education Vol 3 No 1 (2014)
Publisher : Journal of Biology Education

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Abstract

Application measures of inquiry conducted by teachers who do not meet the basic standards, so that students have trouble doing scientific work skills well. Teachers had not found the right method for learning the material Natural Phenomena. In addition, teachers do not monitor students affective value in a sustainable manner, while the affective value determines the success of student learning. This study aimed to analyze the application of scientific inquiry teaching model can improve student affective value and learning outcomes class VII in SMP N 2 Semarang. This research was classroom action research which consisted of 3 cycles of learning. The sample in this research were class VII D and VII E. Results of the analysis of students affective learning value showed the average scientific interests and attitudes of students  in learning increases in each cycle. Average learning outcomes student of class VII D and VII E on the first cycle of 46,32; second cycle of 49,63; and the third cycle 82,81. Conclusions from this research was the application of scientific inquiry teaching model can increase the value of affective and cognitive learning outcomes matter Natural Phenomena class VII D and E in the Junior High School 2 Semarang Penerapan langkah-langkah inkuiri yang dilakukan oleh guru tidak memenuhi standar yang baku, sehingga siswa kesulitan melakukan ketrampilan bekerja ilmiah dengan baik. Guru belum menemukan metode yang tepat untuk pembelajaran materi Natural Phenomena. Selain itu, guru belum memantau nilai afektif siswa secara berkelanjutan, sedangkan nilai afektif menentukan keberhasilan belajar siswa. Tujuan penelitian ini adalah analisis penerapan model pembelajaran scientific inquiry untuk meningkatkan kemampuan afektif dan hasil belajar siswa kelas VII di SMP N 2 Semarang. Penelitian ini merupakan penelitian tindakan kelas dengan kolaborasi antara guru dengan mahasiswa yang terdiri dari 3 siklus pembelajaran. Sampel dalam penelitian ini yaitu kelas VII D dan VII E. Hasil analisis nilai afektif siswa terhadap pembelajaran menunjukkan rata-rata minat dan sikap ilmiah siswa dalam pembelajaran meningkat di setiap siklusnya. Rata-rata hasil belajar siswa kelas VII D dan VII E pada siklus I sebesar 46,32; siklus II 49,63; dan siklus III 82,81.  Simpulan dari penelitian ini adalah penerapan model pembelajaran scientific inquiry dapat meningkatkan nilai afektif dan hasil belajar kognitif materi Natural Phenomena siswa kelas VII D dan E di SMP Negeri 2 Semarang.
MicroRNA-21 as a biomarker for ovarian cancer detection Kartika, Aprilia Indra; Chasanah, Siti Nur; Fitriawan, Akbar Satria; Tanjung, Dewi Sahfitri; Trirahmanto, Addin; Pradjatmo, Heru; Aryandono, Teguh; Haryana, Sofia Mubarika
Indonesian Journal of Biotechnology Vol 23, No 1 (2018)
Publisher : Universitas Gadjah Mada

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Abstract

Ovarian cancer is a lethal disease. One of the problems faced by patients with ovarian cancer is the lack of symptoms in its early stages, which results in it only being detected when it is at an advanced stage. Therefore, there is an urgent need for biomarkers that can predict ovarian cancer precisely. The purpose of this study was to determine the expression of microRNA-21 as a predictive biomarker candidate in both early- and advanced-stage ovarian cancer. This was a cross-sectional study using the blood plasma of 21 healthy control subjects and 37 blood plasma samples from patients with ovarian cancer. Blood plasmas were collected, from which the RNA was isolated. Based on the RNA, the cDNA was synthesized and run through qPCR, the results of which were analyzed using the Livak method. The results showed an upregulation of microRNA-21 in the advanced stage by 2.14 fold compared with the early stage, and 6.13 fold compared with the healthy controls (p < 0.05). The upregulation of microRNA-21 in early-stage ovarian cancer was 2.86 fold compared with the healthy control subjects (p < 0.05). In addition, there was an increase in the expression of microRNA-21 in ovarian cancer by 4.14 fold compared with the healthy controls (p < 0.05). Based on these results, it can be concluded that the expression of microRNA 21 upregulated with the severity of the disease.
OPTIMASI ANNEALING TEMPERATURE PRIMER MRNA RECK DENGAN METODE ONE STEP QRT-PCR Kartika, Aprilia Indra
Jurnal Labora Medika Vol 2, No 1 (2018): Jurnal Labora Medika
Publisher : Jurnal Labora Medika

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Identifikasi mRNA untuk mengukur ekspresi suatu gen dapat dijadikan sebagai alat diagnostik suatu penyakit. mRNA RECK menghasilkan protein RECK yang terbenam dalam membran glikoprotein sel sehingga memperkuat integritas sel. Ekspresi mRNA RECK dikaitkan dengan proses fisiopatologis dalam kanker ovarium.Metode one step qRT –PCR sangat sensitif dan akurat, namun membutuhkan kompleksitas yang tinggi. Salah satu faktor yang dapat mempengaruhi hasil kuantifikasi mRNA adalah suhu annealing primer mRNA RECK.Penelitian ini melakukan optimasi suhu annealing secara in-silico melalui http://tmcalculator.neb.com/ dan http://www.biophp.org/minitools/melting_temperature diperoleh suhu 49°C (57°C primer forward; 54°C primer reverse) dan 51,8°C primer forward; 51,1°C primer reverse. Proses selanjutnya yaitu melakukan running qRT-PCR menggunakan suhu bertingkat. Suhu 55,6°C pada awal optimasi menunjukkan hasil yang baik terlihat dari kurva meltpeak yang spesifik pada suhu 75°C. Namun setelah proses running menggunakan sampel yang berbeda, hasil meltpeak tidak spesifik. Proses optimasi kedua diperoleh suhu 52,5°C dengan meltpeak spesifik suhu 76,5°C. Optimasi annealing dilanjutkan dengan melakukan persamaan konsentrasi RNA template menggunakan pengenceran perbandingan 1:3 (RNA: Nuclease Free Water (NFW)) Kesimpulan dari penelitian ini diperoleh suhu annealing mRNA RECK sebesar 52,5°C dan pengenceran 1:3 RNA dengan NFW, serta penggunaan reagen KAPA one step PCR tanpa penambahan dUTP dan ROX high serta ROX low.
Analisis Profil Protein Daging Kerbau dengan Variasi Konsentrasi Garam serta Pengasapan Berbasis SDS-Page La’lang, Marselaonety; Darmawati, Sri; Kartika, Aprilia Indra
Jurnal Labora Medika Vol 2, No 2 (2018): Jurnal Labora Medika
Publisher : Jurnal Labora Medika

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Daging kerbau memiliki nilai gizi protein dan susunan asam amino lengkap. Kandungan air dan protein yang tinggi menyebabkan daging mudah busuk, sehingga perlu dilakukan pengawetan penggaraman serta pengasapan. Protein daging kerbau dapat dipengaruhi oleh pengolahan bahan pangan, seperti kadar pemberian garam dan pengasapan. Tujuan penelitian untuk menganalisis profil protein daging kerbau dengan variasi konsentrasi garam  10%b/b, 20% b/b, 30% b/b, 40% b/b penggaraman 3 jam serta pengasapan 2 jam. Profil protein daging kerbau dapat dianalisis menggunakan metode SDS – PAGE 12%. Hasil penelitian dari profil protein daging (kontrol), penggaraman 3 jam konsentrasi garam 10%, 20%, 30% dan 40% berturut – turut 28 sub unit protein, 26 sub unit protein,  25 sub unit protein, 23 sub unit protein, dan 21 sub unit protein, sedangkan daging yang diasapkan tanpa garam, daging penggaraman 3 jam serta pengasapan 2 jam konsentrasi garam 10%, 20%, 30% dan 40% berturut -turut 22 sub unit protein, 24 sub unit protein, 20 sub unit protein, 13 sub unit protein dan 12 sub unit protein. Berdasarkan hasil penelitian ini dapat disimpulkan bahwa semakin tinggi konsentrasi garam maka kandungan protein yang terdapat dalam daging kerbau akan rusak dan semakin sedikit hal ini menyebabkan denaturasi protein yang ditandai dengan berkurangnya sub unit protein. Namun protein daging kerbau lebih banyak rusak apabila diasapkan.
Peningkatan Stabilitas Penggunaan Berulangd an Stabilitas Termal Enzim Lipase melalui Imobilisasi pada Zeolit Alam Wardoyo, Fandhi Adi; Kartika, Aprilia Indra
Jurnal Labora Medika Vol 2, No 1 (2018): Jurnal Labora Medika
Publisher : Jurnal Labora Medika

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Thermal stability and lifetime stability for immobilization lipase on zeolit by absorption technique has been carried out. Immobilization of lipase on zeolit was performed via physical adsorption. This reseacrh aimed to determine the effectiveness of pancreatic lipase immobilization in term of thermal and lifetime stability of the hydrolysis activity. The result of this research proved that pancreatic lipase was able to immobilized to zeolit by using the physical adsorption. Immobilized lipase shows better lifetime stability than the free lipase. Immobilized enzyme catalyzed reaction has a lower decreases free fatty acid than that of the free enzyme. Lower decline of value suggests the enzyme is more stable for repeated use. The immobilized lipase was able to maintain its activity after multiple-uses up to four reaction cycles. Similar trend was also observed for thermal stability study. Immobilized lipase shows a higher thermal stability that that shown by free lipase. Immobilized lipase on zeolit was also able to maintain its activity at 40oC.