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Abstrak :
PCSK9 (Proprotein Convertase Substisilin Kexin 9) diketahui berfungsi dalam memetabolisme lipid dalam mendegradasi LDLR (Low Density Lipoprotein Receptor). Penelitian ini dilakukan untuk mengembangkan model hewan tinggi PCSK9 karena terbatasnya model in vivo PCSK9 yang menggambarkan fisiologis manusia. Pada pankreas diketahui memiliki PCSK9, yang berfungsi dapat mempengaruhi sekresi insulin. Insulin merupakan hormon yang diproduksi di beta sel yang terdapat di pankreas dan berfungsi dalam meregulasi gula darah. Pada penelitian ini dilakukan evaluasi efek pemberian HFD (High-Fructose Diet) terhadap peningkatan kadar PCSK9 pada plasma dan pankreas, menggunakan metode uji ELISA dan Western Blot, Kadar PCSK9 pada plasma tikus Wistar (Rattus Novergicus) jantan pada durasi induksi 3 minggu dan 4 minggu terdapat perbedaan yang signifikan (p>0,005), sedangkan kadar PCSK9 pada pankreas berkorelasi positif antara durasi induksi fruktosa dan kadar PCSK9, dimana terjadi peningkatan bersamaan dengan lamanya pemberian HFD berdasarkan analisis statistik peningkatan tersebut tidak signifikan, dengan kadar rata-rata tertinggi pada plasma sebesar 1195,01 ng/ml pada durasi 4 minggu dan pada pankreas memiliki kadar 1029,88 ng/ml pada durasi 5 minggu setelah pemberian HFD. Hasil western blot yang dilakukan pada sampel pankreas tidak dapat dilakukan metode kuantifikasi lebih lanjut, meskipun demikian hasil kualifikasi β-actin memberikan hasil dengan absorbansi tertinggi pada HFD 1 (A= 433,45). ......PCSK9 (Proprotein Convertase Substicilin Kexin 9) is known to function in lipid metabolism in degrading LDLR (Low Density Lipoprotein Receptor). This research was conducted to develop a tall animal model for PCSK9 due to the limited in vivo PCSK9 model that describes human physiology. The pancreas is known to have PCSK9, which functions to affect insulin secretion. Insulin is a hormone produced in beta cells in the pancreas and functions to regulate blood sugar. In this study, we evaluated the effect of HFD (High-Fructose Diet) administration on increasing PCSK9 levels in plasma and pancreas, using the ELISA and Western Blot test methods. PCSK9 levels in plasma of male Wistar rats (Rattus novergicus) at induction duration of 3 weeks and 4 weeks showed a significant difference (p>0.005), while PCSK9 levels in the pancreas positively correlated between the duration of fructose induction and PCSK9 levels, where there was an increase along with the duration of HFD administration based on statistical analysis the increase was not significant, with the highest average plasma level of 1195.01 ng/ml for 4 weeks duration and in the pancreas having levels of 1029, 88 ng/ml for a duration of 5 weeks after HFD administration. The results of the western blot performed on the pancreas sample could not be carried out by further quantification methods, even though the results of the qualification of β-actin gave the result with the highest absorbance in HFD 1 (A = 433.45).

Abstrak :
"ABSTRAK
" Lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV sebagai alternatif agen transfeksi nonviral yang baru telah berhasil disintesis dan diuji karakteristiknya. Penelitian dilakukan dengan tujuan untuk mengetahui karakteristik lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV dalam membentuk kompleks dengan DNA, mengondensasikan DNA, melindungi DNA dari degradasi enzim DNAse, serta mengetahui sitotoksisitas dan kemampuan transfeksinya pada galur sel mamalia, CHO-K1 dan HepG2. Karakteristik lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV kemudian dibandingkan dengan karakteristik dua agen transfeksi komersil, yaitu Poly-L-Lysine dan Lipofectamine 2000. Hasil penelitian menunjukkan bahwa lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV mampu membentuk kompleks dengan DNA pada rasio muatan 0,7, melindungi DNA dari degradasi DNAse pada rasio muatan 1,5, serta memiliki nilai IC50 sebesar 2,940 x 10-2 ?M pada sel CHO-K1 dan 1,964 x 10-2 ?M pada sel HepG2. Namun, lipopeptida tersebut kurang baik dalam mengondensasikan DNA dibandingkan Poly-L-Lysine dan Lipofectamine 2000. Berdasarkan hasil uji transfeksi, lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV telah berhasil mentransfeksikan plasmid pCSII-EF-AcGFP ke dalam sel CHO-K1 dan HepG2. Keberhasilan tersebut ditandai dengan adanya pendaran hasil ekspresi Green Fluorescent Protein pada kedua sel. Meskipun demikian, pendaran yang dihasilkan dinilai kurang maksimal dibandingkan dengan hasil transfeksi oleh Lipofectamine 2000. Hasil transfeksi lipopeptida Pal-CKKHH-YGRKKRRQRRR-PKKKRKV yang optimal berada pada rasio muatan 7 dengan waktu inkubasi 48 jam.

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ABSTRACT
Lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV as a novel nonviral transfection agent has been successfully synthesized and tested by its characteristics. The study aimed to determine the characteristics of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV in forming complexes with DNA, condensing and protecting DNA from degradation of DNAse enzymes, as well as to reveal its cytotoxicity and ability as a transfection agent in mammalian cell lines, CHO K1 and HepG2. The characteristics of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV were compared with the characteristics of two commercial transfection agents, Poly L Lysine and Lipofectamine 2000. The result has showed that lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is able to complex DNA at charge ratio 0.7, protect DNA from DNAse degradation at charge ratio 1.5, and has an IC50 value of 2.940 x 10 2 M in CHO K1 and 1.964 x 10 2 M in HepG2 cells. However, the lipopeptide is not better in condensing the DNA than Poly L Lysine and Lipofectamine 2000. Furthermore, based on the transfection test result, lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV has successfully transfected pCSII EF AcGFP plasmids into CHO K1 and HepG2 cells. The accomplishment is depicted by presence of Green Fluorescent Protein expression in both cells. However, the fluorescence of transfection by lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is not maximal, compare to Lipofectamine 2000. The optimal transfection result of lipopeptide Pal CKKHH YGRKKRRQRRR PKKKRKV is at charge ratio 7 and 48 hours incubation time.

Abstrak :
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