Karakter Biokimia Dan Profil Protein Yogurt Kambing PE Difermentasi Bakteri Asam Laktat (BAL)

Yogurt merupakan salah satu makanan fermentasi dari susu dengan penambahan Bakteri Asam Laktat (BAL). Tujuan dari penelitian ini untuk mengetahui karakter biokimia dan profil protein yogurt kambing PE difermentasi BAL. Susu kambing dan sapi di perah pada pagi hari dan dibagi menjadi 5 golongan: susu segar (sapi dan kambing), susu fermentasi kultur tunggal dengan starter bakteri L. acidophilus, kultur ganda dengan starter bakteri L. acidophilus+S. thermophilus, dan kultur campuran dengan starter komersial (yogurt mix). Protein susu dan yogurt diisolasi dan dimurnikan dengan 5x volume ekstrak buffer lisis (4mM PMSF, 1x PBS, 0,05 % Tween 20) dan diekstraksi dengan sonikasi amplitudo 20%. Separasi pita protein dengan SDS-PAGE discontinous separating gel 15% dan analisis hasil elektroforesis dihitung berat molekulnya berdasarkan protein standar menggunakan Rf. Analisis densitas profil protein menggunakan software Quantity One dan SPSS 15.0. Hasil menunjukkan bahwa pada susu sapi dan kambing segar, kultur tunggal dan ganda, serta yogurt mix ditemukan Κ-casein, β-casein, dan α-S1 casein pada berat molekul antara 30-38 kDa. Sedangkan pada susu kambing segar dan yogurt mix pada berat molekul 36 kDa yaitu α-S2 casein. Secara umum komposisi protein antara susu sapi dan susu kambing adalah sama, tetapi masing-masing memiliki pita protein yang berbeda, sehingga diduga memiliki fungsi yang berbeda pula

Pengaruh Sari Seduh Teh Hitam (Camelia Sinensis) Terhadap Penghambatan Ppar Γ Sel Adiposa Jaringan Lemak Visera Rattus Norvegicus Strain Wistar

Penelitian bertujuan mengetahui pengaruh sari seduh teh hitam terhadap pengekspresian PPAR ã sel adiposa jaringan lemak visera Rattus norvegicus strain Wistar. Duabelas ekor tikus (Rattus norvegicus strain Wistar) jantan umur 6-8 minggu, berat 200 gram, diberi 4 macam perlakuan, yaitu A (diet tinggi lemak + SSTH 0 g/hari), B (diet tinggi lemak + SSTH 0,015 g/hari), C (diet tinggi lemak + SSTH 0,030 g/hari) dan D (diet tinggi lemak + SSTH 0,045 g/hari) selama 90 hari. Setelah masa perlakuan, tikus dibedah dan diambil lemak viseranya. Jaringan tersebut kemudian dibuat preparat dengan metode parafin. Jumlah sel yang mengekspresikan PPAR ã dianalisis dengan menggunakan pewarnaan immunohistokimia dan untuk mengkonfirmasi bentuk sel adiposa digunakan pewarnaan Hematoxylene&Eosin. Antibodi primer yang digunakan adalah anti PPAR gamma poliklonal antibody rabbit IgG dan antibodi sekunder biotin-goat-anti rabbit IgG. Hasil penelitian menunjukkan jumlah sel adiposa yang mengekspresikan PPAR ã mengalami penurunan seiring dengan penambahan dosis SSTH. Hal tersebut mengindikasikan SSTH dapat menurunkan pengekspresian PPAR ã pada sel adiposa jaringan lemak visera

Production And Potency Of Local Rambutan At East Java AS A Candidate Phytopharmaca

Rambutan is a tropical fruit that grow well in Indonesia and the peel is considered as waste. Many researchers' showed that rambutan peel contains polyphenol that could be expected to avoid obesity. The objective of this study was to explore the increasing production of local rambutan and to identify the promising phytochemical compounds on its peel as phytopharmaca candidate against obesity. Survey was conducted on the production of rambutan, potential plantation area, and marketing. Sample of rambutan peel collected from the sub-district Kanigoro, Blitar. Phytochemical compounds were analyzed using TLC, HPLC and FT-IR. Bioassay analysis used obesity rat models. The survey result showed a mean of rambutan production increased 2,6% in 2007-2012. Average production of rambutan 70-120 kg/tree. Vegetative multiplication usually done to maintenance of rambutan quality. The main compound of Rambutan peel extract (RPE) is flavonoids, tannins, ellagic acid and the major functional group of CH3, aliphatic CH3, and C=O. These compounds have a potential activity against obesity. RPE 30 mg/kgBW dose was significantly inhibit the weight gain of obese rats and reducing the adipocyte size (p<0.05)

Cloning and Expression of hGAD65 Gene in E. Coli BL21

The aim of this study is to construct the hGAD65 gene and to identify the hGAD65 clone by using PCR & RFLP. The samples were derived from normal person & DM patient’s blood. Blood DNA was isolated by salting out method and then amplified by PCR with a pair of specific primer, GAD65-F-BamH1-807 & GAD65-R-Xho1-945. The PCR-product was cloned into vector pET-28a and the pET28a-hGAD65-clone was transformed into E.coli BL21 competent cells. The pET28a-hGAD65-clone was confirmed by PCR and RFLP by BamH1 & XhoI. The PCR product of pET28a-hGAD65-clone was one band of 159bp and has two bands 5.3 kb and 159 bp by RFLPwith both restriction enzymes. The GAD65 protein is expressed in 65kD of pET28a-hGAD65-clone. PET28a-hGAD65-clone was able to recognize by gold standard monoclonal antibody specifically. These results indicated that the hGAD65 gene inserted into pET28a properly and provided the GAD65 protein expression. Key words: hGAD65, PCR, pET-28a, RFL

The Amelioration of T2DM Rat Femoral Bone Achieved by Anti-Osteoporosis of Caprine CSN1S2 Protein Through Bone Morphogenetic Protein Signaling Pathway

We investigated the potential anti-glycation and anti-osteoporosis properties of Caprine milk CSN1S2 protein on the serum AGEs and sRAGE level, osteogenic factors expressions, femoral bone mesostructure, histomorphometry, and hydroxyapatite crystals changes in T2DM rats. Varying doses of Caprine milk CSN1S2 protein (0, 375, 750, and 1500 mg/kg BW) were used to treat the control and T2DM rats. We measured AGEs and sRAGE level; RUNX2, OSX, BMP2, and Caspase-3 expressions in rats using ELISA and immunohistochemistry, respectively. The mesostructure and histomorphometry of femoral bone was analyzed using SEM Microscope and BoneJ software, then hydroxyapatite crystal size was determined using SEM-XRD. T2DM rats showed a high level of AGEs and a low level of sRAGE, the RUNX2, OSX, and BMP2 expression was down regulated, BV, BV.TV, Tb.Th, Tb.Sp, increased and SMI levels declined, respectively. Vice versa, after administration of the CSN1S2 protein to T2DM rats, improvement in all levels of molecular and cellular markers was achieved. In the CSN1S2 highest dose, AGEs level declined and sRAGE level elevated in T2DM rats. The 375 and 750 mg/kgBW of CSN1S2 protein was able to upregulate the RUNX2, OSX, and BMP2 expression in T2DM rats, thus improving the normalization of osteoclasts and osteoblasts number. The whole dose of CSN1S2 triggered the thickening of trabecular bone wall, granule formation, and normalized the trabecular thickness (Tb.Th) parameter of T2DM rats. The hydroxyapatite crystal size was increased in the highest dose of CSN1S2-treated T2DM rats. This study indicated that CSN1S2 protein had a protective effect against osteoporosis in the T2DM rat bones by means of glycation pathway inhibition, bone histomorphometry and mesostructure improvement via bone morphometric protein signaling

The Inhibitory Effect of Rambutan Peel Extract to the Igf-1 and Igf-1r Expression in Obese Rats Visceral Fat

The activation of Insulin Like Growth Factor-1 (Igf-1) to its receptor (Igf-1r) plays an important role in the process of adipogenesis. The inhibition of Igf-1 and its receptor at the early process is the precise step for the therapy of obesity. This study used obese rats treated with ellagic acid and Rambutan Peel Extract (RPE) 30mg/kgbw dose. The Igf-1 and Igf-1r expression in visceral fat tissue was observed using immunofluorescence double staining, which confirmed later by western blotting. The results reveal that the use of RPE30mg/kg BW on obese rats model can provide inhibitory effect on the expression of igf-1 and igf-1r as in a normal rat. The expression of igf-1 and its receptor highest in obese rats and the lowest expression in obese rats which were treated with ellagic acid. The reduced expression of igf-1 and its receptor will reduce ERK1/2 and PPAR expression in adipogenesis signaling cascade and in turn inhibits adipogenesis