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Polyphenols extracted from the Green Tea (Camellia sinensis) augments the protective immune responses in mice challanged with Salmonella typhimurium Ratnaningsih, Tri; Asmara, Widya; Sismindari, Sismindari
Medical Journal of Indonesia Vol 13, No 1 (2004): January-March
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (333.117 KB) | DOI: 10.13181/mji.v13i1.122

Abstract

Green tea is an aqueous infusion of dried unfermented leaves of Camellia sinensis. Numerous biological activities of green tea have been reported. The aqueous infusion and its polyphenolic substance are known for their activity as an antimutagenic,  antibacterial, hypocholesterolemic, antioxidant, and mutagenic of B lymphocyte. Studies have demonstrated that green tea polyphenols increase IL-12 production. Salmonella spp infection is an important public health problem in many countries. Cell-mediated immunity (CMI), especially T-cell help is important for protection against this infection. Recent evidence indicates that IL-12 is one such factor that plays a crucial role in the development of CMI. These studies were carired out to investigate the effect of green tea polyphenols to the immune cellulare in mice responses of mice during Salmonella typhimurium infection. The subject consisted of 36 female mice (Balb/C), 6-8 weeks old, divided into 3 groups. The first group was given 10 mg polyphenols/mouse, the second group was given 5 mg polyphenols/mouse, and the third group as the control. In day 31, all mice were infected with 108 CFU Salmonella typhimurium orally. On day 0, 3, 5, and 7 postinfection, 3 mice from each groups were sacrificed, the splenocytes were extracted and cultured to measure  the level of IFN-g in the supernatan and. The peritoneal macrophages were also extracted and cultured to measure the phagocytic activity. The level of IFN-g in splenocyte culture supernatant  increased during infection  in all groups, but the level of the experimental groups  were higher than in control group. The  percentage of phagocytic activity of peritoneal macrophages were higher in the experimental groups than in the control group. The increase of the phagocytic activities were seen corelate with the level of IFN-g supernatan splenocyte culture. (Med J Indones 2004; 13: 1-7)Keywords: polyphenols, green tea, macrophages, phagocytosis
Analisis Pertumbuhan Kartilago Epifisialis Os Tibia Fetus Mencit (Mus musculus L.) Swiss Webster Setelah Induksi Ochratoxin A Selama Periode Organogenesis Setiawan, Arum; Sagi, Mammed; Asmara, Widya; Istriyati, Istriyati
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Publisher : Jurnal Biologi Papua

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Abstract

The aims of this study were determined the effects of Ochratoxin A (OTA) on growth of fetus tibia epiphyseal cartilage during organogenesis period. Twenty four pregnant mice were divided randomly into 4 groups of 6. Ochratoxin A was dissolved in sodium bicarbonateand administered orally on seventh to fourteenth days of gestation at dosage of 0.5, 1.0, 1.5 mg/kg bw, respectively. The remaining were used as control. The fetal tibia was taken after the 18 th day of pregnancy. The growth of tibia epiphyseal cartilages were observed histologically using Erlich’s Haematoxylin-Eosin Stain. The result of this study indicated that OTA caused decreased thickness of the rest zone, proliferative zone, maturation zone and calsification zone of the fetus tibial growth plate significantly.Key words: Ochratoxin A, tibia, cartilage, and thickness
Pertumbuhan dan Perkembangan Otak Fetus Mencit Setelah Induksi Ochratoxin A Selama Periode Organogenesis Setiawan, Arum; Sagi, Mammed; Asmara, Widya; Istriyati, Istriyati
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Publisher : Jurnal Biologi Papua

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Abstract

This experiment was performed to examine the effects of Ochratoxin A (OTA) during organogenesis period on fetuses brain growth and development. Thirty pregnant mice were divided randomly into 5 groups of 6. Ochratoxin A was dissolved in sodium bicarbonate and administered orally on seventh to fourteenth days of gestation at dosage of 0.5, 1.0, 1.5 mg/kg bw, respectively. The remaining animals were used as  an untreated control, and placebo were given by  Sodium Bicarbonate. At the age of 18-day pregnancy, mice were sacrified dan taken its brains. The fetuses brain growth and development  were observed by measure brain weight, cereberum width and length, cerebellum width and length, and the wall thichkness of cerebrum. Result of this studies indicated that OTA caused decreased of brain weight, the length and width of cerebrum and cerebellum, the wall thickness of cerebrum significantly.Key words: Ochratoxin A, brain, cerebrum, cerebellum, mice.
diversity of myiasis fly, Chrysomya bezziana population in Indonesia based on mitochondrial DNA Wardhana, April H; Muharsini, S; Asmara, Widya
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 2 (2004)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (217.954 KB) | DOI: 10.14334/jitv.v9i2.416

Abstract

Chrysomya bezziana is a fly causing myiasis in most livestock in Indonesia. To date, the genetic diversity of C. bezziana has been argued among researchers. The aim of this study was to analyse genetic diversity of C. bezziana population in Indonesia using mitochondrial cytochrome b gene (cyt b) as a marker. The preliminary study showed that the larvae instar III stadium (L3) was the most appropriate sample for molecular analysis and identification. Twenty-four L3 were collected from cattles, buffalos and horses in Bogor, Makassar and East Sumba. DNA samples were isolated from muscle tissue of the larvae. The fragment of cyt b gene (279 bp) was amplified using primer CB3FC-NINFA. The PCR product was subsequently purified, sequenced and analysed using PAUP version 4. The results showed that C. bezziana from Indonesia was different with those from Asia, basicaly for DNA sequences, however it had similar to those from Papua New Guinea. Two haplotypes were identified i.e. haplotype 6 (Makassar haplotype that similar to those from Papua New Guinea) and haplotype 7 (Bogor haplotype that similar to those from Makassar and East Sumba). Key words: Chrysomya bezziana, genetic diversity, mitochondrial DNA
Sequence variability of the β Tubulin isotype-1 genes in benzimidazole resistant strains of Haemonchus contortus, a nematode parasite of sheep Haryuningtyas, Dyah; Artama, wayan T; Asmara, Widya
Indonesian Journal of Animal and Veterinary Sciences Vol 11, No 3 (2006)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (223.925 KB) | DOI: 10.14334/jitv.v11i3.531

Abstract

Resistance to benzimidazole at a population of gastrointestinal nematode, especially Haemonchus contortus is a widespread problem. Therefore, anthelmintic will not effective anymore. Benzimidazole resistance in nematode has been shown to be heritable and defined as an increase in frequency of individual worms able to drug tolerate. In Indonesia resistance to anthelmintic albendazole has been reported on sheep in district of Bogor, Kuningan, Kendal and province of Yogyakarta. Molecular mechanism of resistance to benzimidazole on nematode and fungi related to single gene which code ß tubulin. The aim of this research is to determine a genetics variability of isotype-1 ß tubulin genes from Indonesian strain of H. contortus which resistant to benzimidazole. Sequencing were carried out at a 520 bp fragment of the isotipe-1 ß tubulin gene of six H. contortus resistant strain from four sheep from SPTD Trijaya farm, district of Kuningan, West Java; UPTD Pelayanan Kesehatan Hewan farm, district of Bantul, Yogyakarta respectively. One H. contortus susceptible strains as a control come from farmer from Cicurug, district of Sukabumi, West Java. Results indicated that there are genetics variability of  fragment isotype-1 ß tubulin genes from H. contortus from UPTD Pelayanan Kesehatan Hewan farm, Bantul, Yogyakarta (H1, H2, and C2). There are no genetics variability in the fragment gene of H. contortus resistant strain from SPTD Trijaya farm, Kuningan, West Java (Kn1, Kn2 and K1). Genetic variability occur between 2 isolate from different location. Key Words: Haemonchus contortus, Benzimidazole, Isotype 1 β-tubulin Gene, Sequencing
Effect of Probiotic Lactobacillus sp. Dad13 on Humoral Immune Response of Balb/C Mice Infected with Salmonella typhimurium Kusumawati, Ika Dyah; Harmayani, Eni; Asmara, Widya
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

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Abstract

An indigenous strain of lactic acid bacterium (LAB) identified as Lactobacillus spp. Dad13 (Dad13), isolatedfrom traditional fermented buffalo milk, was found to be potential as probiotic. The aim of this research was to studythe effect of probiotic Dad13 on humoral immune response of Balb/C mice infected with Salmonella typhimurium. Thespecific objective was to find out the effect of different Dad13 consumption time (before and along with infection of S.typhimurium) on the humoral immune response of Balb/C mice. The experiment was conducted by in vivo trial on 20males of Balb/C mice, age of 6-8 weeks, fed with AIN-93 standard diet. The mice were assigned into 4 groups. Eachgroup received the following treatments, ie. Dad13 only, Dad13 before infection, Dad13 along with infection andSalmonella infection only. A volume of 100 μl Dad13 cell suspensions (1010 CFU/ml) were given by oral forced feedingdaily for a week, at week 3 for group before infection and at week 4 for group of Dad13 only and Dad13 along withinfection. Salmonella infection (109 CFU/ml) was given once orally at week 4 to all groups except group treated withDad13 only. The humoral immune response of Balb/C mice was detected 2 weeks after infection by measuring thetiters of IgG and IgA specific from serum and mucosal intestinal liquid samples using Enzyme-linked ImmunosorbentAssay (ELISA) method. The result indicated that humoral immune response of Balb/C mice consuming Dad13 beforeand along with Salmonella infection were significantly different (p<0.05). Dad13 consumption along with Salmonellainfection increased circulated IgG and IgA as well as secretory IgA. It can be concluded that Dad13 probiotic feedingalong with infection increased humoral immune response more significantly compared to that before infection.Key words : Probiotic, Lactobacillus sp. Dad13, Immune response, Salmonella typhimurium
Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant Rintiswati, Ning; Wibawa, Tri; Asmara, Widya; Soebono, Hardyanto
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

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Abstract

AbstractMycobacterium tuberculosis strains resistance to INH is mainly caused by the alteration in several genesencoding the molecular targets. Mutation of katG at codon 315 especially Ser315Thr are responsible forINH resistance in a large proportion of TB cases. The aim of this study is to evaluate the influence of stressoxidative on AhpC activity of katG Ser315Thr of M.tuberculosis, and to find out the relation of AhpC and thevirulence of this mutant. The study design was laboratoric experimental, subjects of study were M.tuberculosisINH resistance strains, and the treatment were serial dose of H2O2. Eighty five M.tuberculosis INH resistantclinical strain were screened for mutation of katGSer315Thr by PCR/RFLP and characterized on the basis ofphenotypic properties (catalase activity and AhpC activity). AhpC activity of katG Ser315Thr M.tuberculosisstrains in response to oxidative stress condition was evaluated by culturing the strains on liquid culturemedium containing 1mM H2O2. To ascertain role of AhpC in the virulence of katGSer315Thr mutant strains, themutants were infected into human macrophages culture, and several indicator of virulence were observed (i.e:replication competence, and apoptosis induction on human macrophages). The results showed that katG Ser315Thr were identified in 23 (27,05%) of 85 INH resistance strains, all mutant strains had decrease of catalaseactivity. AhpC activity of katG Ser315Thr of M.tuberculosis increased significantly with increase of hydrogenperoxide dose. In addition , it has been shown that increased AhpC activity related to replication ability ofmutant, and reduction of apoptosis macrophages induction significantly. We conclude that the productionof AhpC of katG Ser315Thr M.tuberculosis induced by oxidative stress. There was a role of AhpC in virulenceof the M.tuberculosis katG Ser315Thr strains by replication capability and macrophages apoptosis.Keywords : katG Ser315Thr Mycobacterium tuberculosis- oxidative stress - AhpC - virulence
Cytotoxic Activity of Tegari (Dianella nemorosa Lam.) Methanol Extract Against HeLa Cells Karim, Aditya Krishar; ., Sismindari; Asmara, Widya; ., Istriyati
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Dianella nemorosa Lam. also known as tegari belonging to the Liliaceae family. This plant has been utilized for Papua traditional medicine as well as anticancer agent. This research examined potential cytotoxic activity of tegari (D. nemorosa) leaves extract against cervical cancer cell line (HeLa). Methanol extract was obtained by extracting the leaves powder using methanol. Extract was then applied into HeLa cell line to find out the cytotoxic activity. MTT [3-(4,5-dimetilthiazol-2-il)2,5-difeniltetrazolium bromida) assay was used to measure the cytotoxic activity. The result indicated that D. nemorosa leaves extract possessed cytotoxic activity in HeLa cell line with IC50 values were 685,69 µg/ml, 506,43 µg/ml and 708 µg/ml at the incubation period of 24, 48 and 72 h respectively. The strongest cytotoxic was showed by methanol extract incubated in 48 h.
Efektivitas Desensitizing Agent dengan dan tanpa Fluor pada Metode in Office Bleaching terhadap Kandungan Mineral Gigi (Kajian In Vitro) Kristanti, Yulita; Asmara, Widya; Sunarintyas, Siti; Handajani, Juni
Majalah Kedokteran Gigi Indonesia Vol 21, No 2 (2014)
Publisher : Majalah Kedokteran Gigi Indonesia

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Abstract

In office bleaching menggunakan hidrogen peroksida 40% sering memberikan efek samping berupa linu baik selama maupun setelah perawatan tersebut dilakukan. Penelitian ini dilakukan dengan tujuan untuk mengetahui pengaruhwaktu aplikasi dan kandungan fluor desensitizing agent pada metode in office bleaching terhadap kandungan mineral gigi. Delapan gigi masing-masing dipotong menjadi 4 bagian, 6 potong digunakan untuk pemeriksaan XRD awal. Gigi dikeringkan dalam oven suhu 50° selama 30 menit diserbuk, diambil 1 mg untuk diperiksa kandungan mineralnya dengan goniometer. Dua puluh empat potong yang lain dibagi dalam 4 kelompok perlakuan. Kelompok I: gigi diaplikasi bahan bleaching 0,5-1 mm hidrogen peroksida 40% selama 1 jam, dicuci, dikeringkan, diikuti aplikasi 0,1 ml desensitizing agent tanpa fluor (CPP-ACP) selama 30 menit, dicuci, dikeringkan. Kelompok II : gigi diaplikasi CPP-ACP 30 menit, dicuci, dikeringkan, dibleaching menggunakan hidrogen peroksida 40% selama 1 jam. Selanjutnya gigi diaplikasi CPP-ACP 30 menit lagi, dicuci, dikeringkan. Kelompok III gigi dibleaching menggunakan hidrogen peroksida 40% selama 1 jam, dicuci, dikeringkan, diikuti aplikasi desensitizing agent yang mengandung fluor (CPP-ACFP) selama 30 menit, dicuci, dikeringkan. Kelompok IV: gigi diaplikasi CPP-ACFP 30 menit, dicuci, dikeringkan, dibleaching menggunakan hidrogen peroksida 40% selama 1 jam. Selanjutnya gigi diaplikasi CPP-ACFP selama 30 menit, dicuci dikeringkan. Sesudah perlakuan, semua gigi dilakukan pemeriksaan kandungan mineral gigi dengan prosedur yang sama. Uji Mann Whitney menunjukkan penurunan mineral paling sedikit terjadi pada kelompok IV (4500). Desensitizing agent mengandung F sebelum dan sesudah perlakuan in office bleaching menunjukkan penurunan mineral paling kecil. ABSTRACT: The Effectiveness of Desensitizing Agent with and without Fluorine in Office Bleaching Method to Tooth Mineral content. Tooth sensitivity arises during or after an in-office bleaching was performed is usually overcome by using desensitizing agent with or without fluor. So far, desensitizing method applied only reduces tooth sensitivity but it had not overcome demineralization problem yet. This study was aimed to determine the influence of an applicationprocess and fluoride containing desensitizing agent in tooth mineral lossEight teeth were divided into four parts. Six specimens were smoothened using agate mortar. One mg of powder was inserted into the sample holder and mounted ongoniometer heads to examine mineral content before treatment using X-Ray Diffraction (XRD). Twenty-four specimens were classified into four groups. In group I, 0,1 ml desensitizing agent without fluoride (CPP-ACP) was applied for 30 minutes after an in-office bleaching using 40% Hydrogen Peroxide was performed, while in group II, CPP-ACP was applied before and after an in-office bleaching was performed. In group III 0,1 ml fluoride containing desensitizing agent (CPP-ACFP) was applied for 30 minutes after in-office bleaching using 40% Hydrogen Peroxide was performed, while in group IV fluoride containing desensitizing agent was applied before and after an in-office bleaching was performed. Finally, all of the specimens were measured in their after-treatment mineral contain. Mann Whitney test showed that the lowest mineral lost was detected in group IV (4500). Fluoride containing desensitizing agent applied before and after an in-office bleaching effectively reduced mineral loss.
Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer Margino, Sebastian; Behar, Chatarina; Asmara, Widya
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Potato Cyst Nematodes (Globodera rostochiensis) is one of the important potato’s pests and caused economic looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN) shell component of egg shell containing chitin (inner layer) and vitelline/protein (outer layer), so the purpose of research was to fi nd out of chitin degrading bacteria for controlling of egg’s PCN by cutting of their life cycle. The results showed that Bacillus sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation using ammonium sulphate precipitation and DEAE-Cellulose ion-exchange chromatography showed a specifi c activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The apparent Km and Vmax of chitinase towards colloidal chitin were 2 mg/ml and 2.2 μg/h, respectively.