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Construction of an EPO (Human-Erythropoietin) Synthetic Gene Through a Recurvise-PCR Method Fuad, Asrul Muhamad; Gusdinar, Tutus; Retnoningrum, Debbie Sofie; Natalia, Dessy
ANNALES BOGORIENSES Vol 12, No 1 (2008): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

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Abstract

Human  erythropoietin (hEPO)  is an  important glycoprotein  in human  that is coded by a single gene named EPO  (eryhtopoietin). EPO  is  a  glycoprotein  hormone  that  promotes  erythropoiesis,  which  is  the  formation process of mature  red  blood  cell  (erythrocytes)  in  human  bodies.  It  is  widely used  for  treatment  of anemia  in patient  With  chronic  renal failure.  Therefore  EPO has  been classified  as hematopoietic cytokine. Recombinant hEPO  (rhEPO)  has  been  commercially  available,  such  a  Epogen.  It  is  produced  in  mammalian  cell, such  as CHO  (Chine  e hamster ovary) cells  for  the  reason of  its complex structure as a glyco-protein. In  an effort  to  use and optimize heterolgous EPO gene expression  in  an  alternative eukaryotic  host  cells  such  as  yeast, an  EPO­synthetic  gene  (EPOsyn)  was  constructed.  The  synthetic  gene  had  been designed to contain  optimaI Pichiapastoris codon usage . It had  been constructed by a  recursive-PCR method  in  two-step PCR reactions. The gene was assembled  from  8 single strands synthetic  oligonuclotides having an average  length of 90 nt with 20  to 30 overlap  region  between  two  adjacent  oligos. The  synthetic  gene  has  less  GC  content  (4-.3 1%)  compared  its native (human) gene (59.08%). The synthetic gene has  been cloned  in  pCR2.1  cloning plasmid and sequenced. From  8  independent clones,  it was revealed  that  the error  rate  was  1.59%,  in which  1.42% was due  to deletions and 0.17% due  to substitutions. Design of  the gene sequences, construction method and DNA sequence analysis of  the gene will  be discussed  in  this  paper.   Keywords: Human erythropoietin (hEPO), erythropoiesis  EPO­synthetic  gene, recursive-PCR, Pichiapastoris, hematopoietic cytokine.
Biochemical research of phenyclidine reseptor : Interaction to specific bonding to the lower affinity receptor Gusdinar, Tutus; Chicheportiche, Robert
INDONESIAN JOURNAL OF PHARMACY Vol 18 No 4, 2007
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

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Abstract

Fixed concentration of MK-801 molecule had been used as a specific ligand for differenciating two kinds of phencyclidine (PCP) and thienylphencyclidine (TCP) receptors. By using several drug molecules which have different pharmacological action, as well as such molecular affinity (IC50) to the rat brain, it was obtained that both of these interacting receptors had equal character for its higher or lower affinity sites. The MK-801 molecule inhibited much strongly (IC50 is about 10 nM) fixation of 3HTCP to its higher affinity receptor and much weakly (IC50 is about 10 μM) fixation of 3H-PCP to its lower affinity receptor. The MK-801 molecule could be used as well for differenciating two 3H-TCP receptors, the higher and the lower affinity.Key words : phencyclidine – thienylphencyclidine – MK-801 – higher affinity receptor– lower affinity receptor.
Synthesis and gastric ulcer protective activity of chlorinated quercetin Gusdinar, Tutus; Herowati, Rina; Kartasasmita, R. E.; Adnyana, I Ketut
INDONESIAN JOURNAL OF PHARMACY Vol 20 No 4, 2009
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

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Abstract

Gastrointestinal toxicity due to non-steroid anti-inflammatory drugs can be inhibited by the compounds that have antioxidant activity. Quercetin is a flavonoid that has antioxidant activity and protection effect against gastric ulcer. Chlorination of quercetin enhanced the antioxidant activity. This study aims to obtain the chlorinated derivative of quercetin and examine the protection effect against acetosal-induced gastric ulcer. Chlorination was done by the addition of chlorine at room temperature. Ulcer induction was carried out on rats by oral administration of acetosal. Incidences of gastric ulcer were determined by macroscopic and microscopic observation. Chlorination of quercetin with chlorine gas produced 6-chloroqueretin as major product. The protection effect against acetosal-induced gastric ulcer of this compound was higher than quercetin.Key words : quercetin, chlorination, gastric ulcer, NSAIDs
Analisis Kuantitatif 15-F2t-isoprostan dari Plasma Pasien Obstructive Sleep Apnea (OSA) dengan Metode Enzyme Linked Immunosorbent Assay (ELISA) menggunakan Teknik Ekstraksi Imunoafinitas Rusdi, Bertha; Gusdinar, Tutus; Fattah, Miswar
Indonesian Journal of Clinical Pharmacy Vol 1, No 1 (2012)
Publisher : Indonesian Journal of Clinical Pharmacy

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Abstract

15-F2t-isoprostan merupakan penanda stres oksidatif yang kadarnya dalam cairan biologis relatif rendah serta memiliki banyak isomer sehingga diperlukan ekstraksi sampel sebelum dilakukan pengukuran kadar. Metode ekstraksi yang digunakan diantaranya ekstraksi fase solid/Solid Phase Extraction (SPE) serta ekstraksi imunoafinitas. Perbaikan teknik ekstraksi SPE dan teknik ekstraksi imunoafinitas dilakukan untuk membandingkan hasil perolehan kembalinya. Pengukuran dilakukan dengan metode Enzyme Linked Immunosorbent Assay (ELISA). Penilaian efektivitas proses ekstraksi diamati melalui hasil perolehan kembali dari kedua teknik ekstraksi. Teknik ekstraksi dengan perolehan kembali tertinggi digunakan untuk mengukur kadar 15-F2t-isoprostan dari penderita Obstructive Sleep Apnea (OSA). Teknik ekstraksi imunoafinitas menghasilkan perolehan kembali 15-F2t-isoprostan yang cukup baik. Pada penderita OSA kadar 15-F2t-isoprostan dalam plasma cenderung tinggi sehingga memiliki risiko untuk menderita penyakit yang berhubungan dengan aktivitas biologis 15-F2t-isoprostan seperti arteriosklerosis.Kata kunci: 15-F2t-isoprostan, Solid Phase Extraction (SPE), ekstraksi imunoafinitas, ObstructiveSleep Apnea (OSA) Quantitative Analysis of Free 15-F2t-Isoprostane from Plasma of Obstructive Sleep Apnea Patients Using Enzyme Linked Immunosorbent Assay Method with Immunoaffinity Extraction TechniqueAbstract15-F2t-isoprostane is a biomarker in assessment of oxidative stress status that due to its relatively low concentration in biological fluid and also has many isomers, the 15-F2t-isoprostane sample need to be extracted prior to the quantifying processes. Extraction techniques commonly used to extract 15-F2t-isoprostane are solid phase extraction (SPE) and immunoaffinity extraction. Improvements to the SPE and immunoaffinity extraction techniques had been conducted, and the recovery results was then compared. The quantification of 15-F2t-isoprostane then was conducted using Enzyme Linked Immunosorbent Assay (ELISA) method. Then followed by the examination of the plasma recovery results. Extraction technique which had the highest recovery then was used to quantify 15-F2t-isoprostane from plasma of Obstructive Sleep Apnea (OSA) patients. Immunoaffinity extraction technique has a good recovery result. OSA patients have the tendency to have high 15-F2t-isoprostane concentrations in the plasma, therefore have a potential risk to get diseases related to the biological activities of 15-F2t-isoprostane, such as arteriosclerosis.Key words: 15-F2t-isoprostane, Solid Phase Extraction (SPE), immunoaffinity extraction, ObstructiveSleep Apnea
Production and Application of Lactobacillus plantarum IBL-2 Bacteriocins as Meat Product Biopreservatives Wibowo, Marlia Singgih; Muzakiyah, Isra; Nurhayati, Betty; Padmasawitri, Tjokorde Armina; Widiastuti, Yantyati; Gusdinar, Tutus
Indonesian Food and Nutrition Progress Vol 14, No 1 (2017)
Publisher : Indonesian Association of Food Technologists

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Abstract

Biopreservation is one of the alternatives to obtain safe food products. The produced bacteriocin by Lactic Acid Bacteria (LAB) is potential as biopreservatives, which is safe for consumption, since it was a protein degradable by proteolytic enzymes. This study aimed to optimize bacteriocin production from L. plantarum IBL-2 and to evaluate the effectiveness of bacteriocins in reducing the number of total plate count and Salmonella typhimurium in ground beef. Bacteriocin was produced through fermentation of L. plantarum IBL-2, under various conditions to yield the compound with the best antimicrobial activity. The total number of bacteria in ground beef after the addition of L. plantarum IBL-2 fermentation supernatant was determined. The result was compared with the sample without preservatives (control), and sample added with commercial Nissin. All three samples of ground beef were spiked with S. typhimurium and incubated for 0, 2, 6, 8, 12, 14 days at a temperature of 4-10 ° C. Total Plate Count (TPC) method was utilized to determine the number of bacteria in the samples. The fermentation process resulted in bacteriocin with the strongest antimicrobial activity when using low molecular weight liquid medium (LMWLM), followed by a series of refining process. From day 0-14, the number of S. typhimurium, in sample added with L. plantarum IBL-2 fermentation supernatant, was lower than control and sample added with Nissin. The most optimal antimicrobial activity of bacteriocin was obtained using LMWLM as fermentation media, and using a series of refining process consist of bacteriocin supernatant evaporation, membrane ultrafiltration, and gradual fractionation using 80% ammonium sulphate. Bacteriocin from L. plantarum IBL-2 showed antimicrobial activity against S. typhimurium.
KAJIAN KOMPOSISI KIMIA, NILAI NUTRISI, DAN ETNOFARMAKOLOGIS TANAMAN GENUS KENARI Rahman, Hamidah Rahman; Anggadiredja, Kusnandar; Gusdinar, Tutus; Sitompul, Johnner P; Ryadin, A R
Jurnal Fitofarmaka Indonesia Vol 6, No 1 (2019): JURNAL FITOFARMAKA INDONESIA
Publisher : FAKULTAS FARMASI UNIVERSITAS MUSLIM INDONESIA

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Abstract

One species of canarium genus is Canarium indicum L which is an original plant from Indonesia. Utilization of some canarium nut species which spread around the world had been recognized, such as a source of nutritious food as well as its potency as raw material of medicine based on evidence-based scientific. In traditional medicine, was reported if canarium nut had been used for diabetes mellitus treatment, angina, hepatoprotective, anti-inflammatory, antirheumatic, expectorant, etc. This article aims to review the chemical composition, nutrient values, and potency of canarium nut as the raw material of medicine. Scientific data were obtained from online search results. From the searching was found that at least eight species of canarium nut reported which is related to chemical composition and its nutrition which also provide ethnopharmacological benefits. Overall results, all identified canarium nut species have a favorable chemical composition and nutritional values that can be utilized as functional, nutritional and medicinal food. Therefore, the result of this study also indicates if the canarium nut from Indonesia, C. indicum L., has not been thoroughly studied. That indicates by the lack of scientific paper which presented about C. indicum from Indonesia.