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Abstrak :
ABSTRAK
Proses pematangan spermatozoa terjadi karena adanya interaksi antara protein dengan membran plasma spermatozoa. Walaupun proses pematangan spermatozoa ini sangat penting, namun gen yang berperan dalam sekresi protein di epididimis ini masih banyak yang belum dikarakterisasi. Gen-gen yang berperan dalam proses pematangan spermatozoa umumnya merupakan protein sekretorik, terekspresi pada segmen spesifik, diregulasi androgen, faktor testikular dan perkembangan postnatal. Pada penelitian sebelumnya diketahui bahwa b-defensin merupakan gen yang banyak terekspresi di organ reproduksi pria dan memiliki peran dalam pertahanan tubuh dan pematangan spermatozoa. Penelitian ini dilakukan untuk mengkarakterisasi ekspresi gen Defb20 untuk mengetahui perannya dalam proses pematangan spermatozoa. Studi in silico dilakukan untuk prediksi struktur gen, signal peptide dan domain fungsional. Quantitative real-time PCR digunakan untuk mengukur ekspresi gen Defb20 pada analisis sebaran jaringan, regulasi androgen dan faktor testikular serta postnatal developmen. Hasil penelitian mendapatkan bahwa sekuen Defb20 mengandung domain penting seperti N-myristoilation dan beberapa situs phosporilasi protein kinase yang mungkin berperan dalam mekanisme interaksi protein dengan membran plasma. Sekuen asam amino Defb20 mengandung signal peptides, mengindikasikan protein yang disekresikan dan terlibat dalam proses pematangan spermatozoa. -defensins 20 (Defb20) terekspresi spesifik di epididimis dengan ekspresi tertinggi terdapat pada kaput epididimis. Defb20 diregulasi oleh androgen yang ditunjukkan dengan adanya penurunan ekspresi Defb20 paska dilakukan gonadektomi dan kondisi ini dapat diperbaiki dengan pemberian hormon pengganti. Defb20 juga diregulasi oleh faktor testikular, yang dibuktikan dari menurunnya ekspresi Defb20 setelah ligasi pada duktus eferen (efferent duct ligation (EDL)). Defb20 mulai terekspresi pada hari ke-21 setelah lahir yang mengindikasikan gen Defb20 terekspresipada suatu periode perkembangan epididimis. Berdasarkan hasil penelitian disimpulkan bahwa Defb20 memiliki karakteristik ekspresi : mengandung signal peptide yang mengarahkan sintesis protein pada jalur sekretorik, spesifik terekspresi di epididimis, diregulasi androgen dan faktor testikular serta mulai terekspresi pada masa pubertas hingga dewasa

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ABSTRACT
Epididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode secreted proteins remain largely uncharacterized. The genes that play a role in sperm maturation process has character, among others; is a secretory protein, expressed specifically in the epididymis, regulated by androgen, testicular factor, and postnatal development. Previous studies showed that family of-defensins preferentially eaxpressed in male reproductive tracts and play an important role in both innate immunity and sperm fertility. This study aimed to characterize Defb20 to understand its role in sperm maturation. This study using in silico analyses and quantitative real-time PCR (qRT-PCR). In silico analyses were performed to predict gene structure, signal peptides and functional domains. Defb20 expression in various tissues, after gonadectomy, efferent duct ligation and postnatal development were measured using quantitative real-time RT-PCR. Defb20 sequence contains important domains such as N-myristoilation and kinase binding sites which are putatively involved in the protein activation and protein-plasma membrane interaction. The amino acid sequence of Defb20 contains signal peptides, indicating characteristic of secretory proteins involved in the sperm maturation. β-defensins 20 (Defb20) was expressed exclusively in the epididymis with the highest expression in the caput region. Defb20 was regulated by androgen showing down-regulation after gonadectomy and the expression was recovered after testosterone replacement. However, Defb20 was also regulated by testicular factors in which the expression was down-regulated after efferent duct ligation (EDL). The dependency on the androgen was further confirmed by postnatal expression analysis in which Defb20 begin to express at day21 postnatal indicating specific stage of expression after initial development of the epididymis. In conclusion, Defb20 have a potential to be involved in the epididymal sperm maturation process. Defb20 has characteristic expression; has a signal peptide sequence that directs synthesis in the secretory pathway, specifically expressed in the epididymis, androgen and testicular factors regulated, and expressed in puberty to adulthood

Abstrak :
Tingkat globalisasi yang tinggi mempengaruhi gaya hidup masyarakat yang cenderung menginginkan serba instan dan menyebabkan diet yang tidak sehat. Estimasi dari International Diabetes Federation menyatakan bahwa pada tahun 2035 diperkirakan akan terdapat 592 juta orang penderita diabetes melitus, dengan di Indonesia menjadi penyebab kematian nomor 3 pada tahun 2017. Pada penelitian ini, dilakukan perancangan produksi obat ekstrak daun sambung nyawa yang memiliki efek anti-hiperglikemik dan ekonomis. Tahapan penelitian ini dibagi menjadi 3 fase, yaitu simulasi interaksi zat aktif daun sambung nyawa terhadap protein penyebab hiperglikemik (11-beta-hidroksisteroid dehidrogenase-1; interleukin-1; dan protein tirosin fosfat), simulasi proses pembuatan obat ekstrak daun sambung nyawa dengan pelarut air, serta pemodelan reaksi enzimatik inhibisi non-kompetitif hasil obat hasil simulasi proses dengan besaran interaksi hasil simulasi interaksi zat aktif. Hasil penelitian memperlihatkan interaksi inhibisi zat aktif daun sambung nyawa (Asam p-hidroksibenzoat, asam p-hidroksinamat, kuersetin) dan obat standar (Metformin) terhadap protein penyebab hiperglikemik menggunakan program Autodock v.4.0 berhasil dilakukan dan memperlihatkan adanya interaksi inhibisi zat aktif terhadap protein. Hasil penelitian juga memperlihatkan simulasi produksi ekstrak daun sambung nyawa dengan pelarut air menggunakan program SuperPro Designer v.9.0 berhasil dilakukan dengan memberikan hasil perkiraan produksi setiap tahunnya 4.133.250 kapsul ekstrak dengan harga Rp 11.000 untuk setiap kapsulnya, komposisi zat aktif yang terdapat pada setiap 1 kapsul, serta menghasilkan rasio keuangan yang layak investasi (ROI sebesar 140,84%; PBP sebesar 1,43 tahun; BEP sebesar 782.459 kapsul; IRR sebesar 91,65%; dan NPV sebesar Rp 51.991.389.985). Kedua hasil penelitian kemudian memberikan hasil penelitian fase ketiga untuk pemodelan reaksi enzimatik inhibisi non-kompetitif dan menghasilkan perkiraan besar aktivitas inhibisi setiap zat aktif dan obat standar terhadap protein penyebab hiperglikemik. ......The effect of high levels of globalization affects the lifestyles of people who tend to want instantaneously and cause an unhealthy diet. Estimates from the International Diabetes Federation state that in 2035 there will be an estimated 592 million people with diabetes mellitus, with Indonesia becoming the number 3 cause of death in 2017. In this research, the design of the production of longevity spinach extract drugs has an anti-hyperglycemic activity and good economic potential. The stages of this research were divided into 3 phases which are the simulation of the interaction of the active substance of the longevity spinach to the target protein that causes hyperglycemic (11-beta-hydroxysteroid dehydrogenase-1; interleukin-1; and protein tyrosine phosphate), the simulation of the process of making the extract of longevity spinach with water solvent, and modeling enzymatic reactions of non-competitive inhibition of the drug which is the results of process simulation with its magnitude of the inhibition interaction results of the simulation of active substance interactions. The results of the study showed that the interaction of the active substances of the longevity spinach (p-hydroxybenzoic acid, p-hydroxycinnamic acid, quercetin) and standard drug (Metformin) to the target protein using Autodock v.4.0 was successfully carried out and showed interactions of inhibition of the active substances against the protein. The results of the study also showed that the simulation of the production of longevity spinach extracts with water solvent using SuperPro Designer v.9.0 was successfully carried out by providing an estimated annual production of 4,133,250 extract capsules for Rp 11,000 for each capsule, the composition of active substances contained in every 1 capsule, and produce a good financial ratio that is worth investment (ROI of 140.84%; PBP of 1.43 years; BEP of 782,459 capsules; IRR of 91.65%; and NPV of Rp. 51,991,389,985). The two research results then provide the third phase research results for modeling the enzymatic reaction of non-competitive inhibitions and produce estimates of the inhibition activity of each active substance and standard drugs against hyperglycemic-causing proteins.