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Abstrak :
Latar belakang: Ribonucleic acid RNA adalah molekul yang tidak stabil secara termodinamik. Cara penyimpanan RNA sangat kritis untuk menjaga kuantitas dan kualitasnya agar dapat digunakan untuk analisis molekuler seperti real time-PCR. Tujuan penelitian ini adalah mengetahui suhu ideal penyimpanan RNA di antara -80°C, -20°C dan 4°C dengan melihat perubahan pada konsentrasi RNA selama dua minggu masa penyimpanan. Metode: Delapan hati tikus dibagi menjadi 3 untuk setiap grup dengan berat masing-masing sampel 25-26 ug. Sampel hati dihomogenisasi dan diisolasi untuk mendapatkan RNA murni, lalu disimpan pada tiga suhu berbeda yakni -80°C, -20°C and 4°C. Absorbasi diukur dengan alat Varioskan Flash pada gelombang cahaya 260 dan 280 nm untuk mendapatkan konsentrasi dan kemurnian sampel. Hasil: Tidak ditemukan perbedaan yang signifikan antara konsentrasi RNA dengan suhu penyimpanaan selama dua minggu, baik secara eksperimental dan secara statistik Kruskal-Wallis, -80°C p = 0.949; -20°C p = 0.885; 4°C p = 0.935 . Dapat disimpulkan bahwa suhu ideal untuk penyimpanan RNA tidak dapat ditetapkan. RNA dengan konsentrasi yang tinggi memiliki kemurnian yang tinggi juga. Kesimpulan: RNA dapat disimpan di suhu -80°C, -20°C dan 4°C selama dua minggu tanpa perubahan kuantitas. Tetapi, durasi studi sebaiknya diperpanjang paling tidak selama satu bulan untuk melihat penurunan pada konsentrasi RNA di suhu penyimpanan yang terkait. Walaupun konsentrasi pada sampel tidak berubah signifikan, kualitas pita RNA tidak dapat dievaluasi untuk analisis molekuler. Analisis kualitas RNA dapat dilakukan untuk melihat terjadinya degradasi.

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Background Ribonucleic acid RNA is a thermodynamically unstable molecule. The way RNA samples are preserved is critical to maintain maximum yield and quality therefore it is useful for molecular analysis such as real time PCR. There are many contradictions and variations regarding the ideal temperature for RNA storage. The aim of this study was to find the ideal temperature for RNA storage among 80°C, 20°C and 4°C by observing for alteration in concentration over two weeks of storage time. Methods Eight mouse liver were each divided into 3 groups, weighed to 25 26 ug. Samples were homogenized and isolated for pure RNA, and were subsequently stored in temperatures of 80o C, 20°C and 4°C. Absorbance was measured with Varioskan Flash photometric tool, at wavelength of 260 and 280 nm. Concentration and purity of RNA samples were then calculated. Results There was no significant difference between concentrations of RNA samples stored in all temperatures across the duration of two weeks storage time, both experimentally and statistically Kruskal Wallis, 80o C p 0.949 20o C p 0.885 4o C p 0.935 . We conclude that the ideal temperature for RNA storage cannot be defined. Higher concentration of RNA coincides with higher RNA purity. Conclusion RNA can be stored in 80o C, 20o C and 4o C for two weeks without quantity reduction. However, longer duration of study, at least one month, is needed to observe whether RNA concentration will be reduced overtime in any of temperatures of storage. Despite the concentration that stayed constant over the duration of storage two weeks , we are unable to determine whether the quality is appropriate for use in molecular assays. Further RNA quality analysis is recommended to check for degradation.

Abstrak :
RNA Interference: Application to Drug Discovery and Challenges to Pharmaceutical Development provides a general overview of this rapidly emerging field, with a strong emphasis on issues and aspects that are important to a drug development team. The first part covers more general background of RNA interference and its application in drug discovery. In the second part, the book addresses siRNA (small interfering RNA), a pharmaceutically potent form, and its use and delivery in therapeutics along with manufacturing and delivery aspects.

Abstrak :
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. ......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.

Abstrak :
Glioma is one of the most common central nervous system tumors that show variant responses towards radiotherapies. Most of the cases especially the high grade Glioblastoma Multiforme have very poor prognosis. Pluripotency of cMyc genes might be another factor for the high glial cell differentiation in glioma thus it may become an alternative therapeutic target. mRNA obtained from 20 glioma samples with different degree of malignancy are converted to cDNA and then amplified. Relative quantification of cMyc mRNA expression is measured by calculating the cycle threshold values of Real Time RT PCR and normalized towards 18s rRNA to predict the relationship between the expression of cMyc and the degree of malignancy. The cMyc expression is increased in accordance with the tumor grade. The cMyc expressions in high grade glioma are 17424.23 folds higher when calibrated to the normal cell, whereas the genes in lower grade tumors are expressed with the rate of 6167.35. Despite the statistically insignificant values the genes express, this research has strengthened molecular diagnosis, specifically pluripotency, to be the factor that gives a greater prognostic relevance than the histopathologic diagnosis. As a conclusion, there is a clinical tendency where the c Myc expression is higher than in high degree glioma compared to low degree malignancy, however it is not statistically significant.

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Glioma adalah salah satu tumor sistem saraf pusat yang sering terjadi dan memiliki respon yang variatif terhadap radioterapi. Glioblastoma Multiforme cenderung memiliki prognosis buruk terhadap pengobatan. Pluripotensi mRNA cMyc dapat menjadi salah satu faktor tingginya diferensiasi sel glial pada gliom sehingga dapat menjadi target terapi alternatif. mRNA yang diperoleh dari 20 sampel glioma dengan derajat keganasan berbeda ditransformasi menjadi cDNA dan diamplifikasi menggunakan Accupower Two-Step RT-PCR with SYBR Green. Kuantifikasi relatif mRNA cMyc ditentukan dengan menghitung nilai cycle threshold pada RT PCR ang dinormalisasi dengan rRNA 18S untuk melihat hubungan antara ekspresi cMyc dan derajat keganasan glioma. Ekspresi cMyc ternyata lebih tinggi seiring dengan meningkatnya tingkat keganasan. Ekspresi cMyc pada glioma klasifikasi WHO derajat tinggi senilai 17424.23 kali lebih tinggi dibandingkan dengan ekspresi pada sel otak normal, sedangkan glioma derajat rendah menurut klasifikasi WHO mengalami ekspresi gen cMyc senilai 6167.35. Meskipun nilai yang diperoleh tidak signifikan secara statistik, penelitian ini telah menunjukkan bahwa diagnosis molekuler, terutama pluripotensi, dapat menjadi faktor penentu prognosis glioma selain ditentukan dengan derajat keganasan melalui pemeriksaan histopatologis. Terdapat kecenderungan secara klinis dimana ekspresi relatif mRNA cMyc lebih tinggi pada glioma derajat tinggi dibandingkan dengan glioma derajat rendah, namun nilainya tidak signifikan secara statistik.

Abstrak :
Some diseases, such as cancer, hereditary and genetic diseases, as well as viral infectious diseases, have been treated unsatisfied by the conventional therapy so far, and even more, by the gene therapy. Together sixth the pharmaceutical industries, researchers put their best effort to hunt some molecules that can be mow favorable for such kind of therapy. After a pivotal study reported in May, 2001, it is certain that Ribonucleic acid (RNA) could effectively silence gene expression in mammalian cell line, SQ it was then proposed in 2004 the term RNA therapeutics. Antisense RNA therapy which came into the atage earlier seemed to be the one that can answer all the problems in knocking out the unwanted messenger in gate expression. RNA interference (RNAi) concept, which came later us around 2QOQ, began to look like a possible contender. It was reported some studies that RNAi seem to have some more advantages over hath stronger gene-silencing effects and greater ease of use. However, the main obstacle of all kind of gene therapy is, undoubtedly, on the delivery of this molecule to enter the target eel!, and mostly to where it is needed most inside the body. Some studies on genetic Material delivery system have been reported, and their progress has been discussed.

Abstrak :
Some diseases, such as cancer, hereditary and genetic diseases, as well as viral infectious diseases, have been treated unsatisfied by the conventional therapy so far, and even more, by the gene therapy. Together with the pharmaceutical industries, researchers put their best effort to hunt some molecules that can be more favorable for such kind of therapy. After a pivotal study reported in May, 2001, it is certain that Ribonucleic acid (RNA) could effectively silence gene expression in mammalian cell line, so it was then proposed in 2004 the term RNA therapeutics. Antisense RNA therapy which came into the stage earlier seemed to be the one that can answer all the problems in knocking out the unwanted messenger in gene expression. RNA interference (RNAi) concept, which came later in around 2000, began to look like a possible contender. It was reported in some studies that RNAi seems to have some more advantages over both stronger gene-silencing effects and greater ease of use. However, the main obstacle of all kind of gene therapy is, undoubtedly, on the delivery of this molecule to enter the target cell, and mostly, to where it is needed most inside the body. Some studies on genetic material delivery system have been reported, and their progress has been discussed.

Abstrak :
This comprehensive volume is intended for readers with research or teaching interests in ncRNA biology and will provide a major information resource on current research in the fast-moving fields of RNA and gene expression regulation.