Indonesian Journal of Biotechnology
Not a member yet
387 research outputs found
Sort by
-429 T/C and -374 T/A Polymorphisms in Receptor Advanced Glycation Endproducts (RAGE) gene in Type 2 Diabetic Patients with Diabetic Retinopathy at the Dr. Sardjito General Hospital Yogyakarta
No full textReceptor of advanced glycation endproduct (RAGE) plays an important role in the pathogenesis of diabetic vascular complications, such as diabetic retinopathy. The interaction between the RAGE and advanced glycation end product (AGE) leads to oxidative stress and could result in cellular activation and infl ammation. The production of AGE occurs normally during aging but it increases in hyperglycemia condition. The objective of this research was to investigate the association between -429 T/C and -374 T/A polymorphisms in RAGE gene with the risk of diabetic retinopathy (DR) of type 2 diabetic patients in Javanese population. This was a case control study which consisted of 40 type 2 diabetic patients with DR as case subjects and 40 type 2 diabetic patients without DR (NDR) as control subjects. Genotyping of polymorphism was performed by PCR-RFLP. Chi-square test and odds ratio models were used to evaluate the association of both polymorphisms and DR risk and to examine 2-SNP haplotype of -429 T/C and -374 T/A polymorphisms in RAGE gene on DR. The genotype frequencies of -429 T/C polymorphism in RAGE gene in DR subjects were TT = 72.5% and TC/CC = 27.5%; while in NDR subjects were TT = 80% and TC/ CC = 20%, with p = 0.431. The allele frequencies of -429 T/C polymorphism in DR subjects were T = 83.7% and C= 16.3%, while in NDR subjects were T = 87.5% and C = 12.5%, with p = 0.499. The genotype frequencies of -374T/A polymorphism in RAGE gene in DR subjects were TT = 67.5%, TA = 32.5% while in NDR subjects were TT =82.5%, TA = 17.5%, with p = 0.121. In DR subjects, the frequencies of T and A were 83.7% and16.3%, while in NDR subjects the frequencies of T and A were 91.2 % and 8.8%, with p = 0.151. Odds ratios of -429 T/C polymorphism were 1.52 (95% CI = 0.54 – 4.29) for TC/CC genotype and 1.358 (95% CI = 0.56 – 3.31) for C allele. Odds ratios of -374 T/A polymorphism were 2.27 (95% CI = 0.79 – 6.49) for TA genotype and 2.02 (95% CI = 0.76 – 5.37) for A allele. χ2-value for 2-SNP haplotype was p = 0.127. The -374 T/A polymorphism in RAGE gene was a stronger risk factor of DR than -429 T/C polymorphism in RAGE gene. There were not signifi cantly different of frequencies of genotypes, allele, and two-SNP haplotype of -429 T/C and -374 T/A polymorphisms in RAGE gene between DR subjects and NDR subjects
Diversity of Dibenzofuran-Utilizing Bacteria Isolated by Direct-Plating and Enrichment Methods
No full textThe effect of enrichment bias on the diversity of Dibenzofuran (DBF)-degrading bacteria recovered from soil was evaluated by direct plating, plating after in-soil adaptation, and plating after batch culture enrichment. Among colonies appeared on Bushnell Haas agar with DBF as the sole carbon source, 119 colonies (49, 38, and 32 from direct plating, plating after in-soil adaptation, and plating after batch culture enrichment, respectively) were arbitrarily selected based on the appearance of the colonies. Total DNA were then extracted from the rest of the colonies and analyzed for their diversity using Ribosomal Intergenic Spacer Analysis (RISA). Number of DNA bands obtained from direct plating was higher than the ones obtained after in-soil enrichment and batch culture enrichment. The RISA bands obtained from direct plating were also found to be distributed more evenly than the ones obtained after in-soil enrichment and batch culture enrichment. Dominant bands were observed on RISA from samples obtained after in-soil enrichment and batch culture enrichment. Out of 119, only 9 isolates were consistently able to grow on Bushnell-Haas broth with DBF as the sole carbon source as indicated by broth turbidity. All of the isolates were obtained from soil samples which were enriched in a batch culture. Some of the isolates were able to degrade more then 80 % DBF in the minimal medium
Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin
No full textExamination of Phalaenopsis amabilis orchid resistance to hygromycin antibiotic is an important step to doprior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposedthe plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilisprotocorms. These protocorms were subcultured onto NP medium supplemented with various concentrationof Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocormsper total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused deathclose to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriateconcentration 10 mg/l, and this concentration can be used for other purposes in orchid system
Human Origin Lactobacillus casei Isolated from Indonesian Infants Demonstrating Potential Characteristics as Probiotics in vitro
No full textThe aim of this experiment was to isolate and identify Lactic Acid Bacteria (LAB) from infant faecesand subsequent evaluation of its potential probiotics. LAB was isolated from faeces of infants who consumedbreast milk as the only source of diet on L-cysteine-supplemented MRS Agar, and incubated on 37oC for 48hours. Colonies grew on this media were then identifi ed based on morphological, physiological and molecularapproaches. Morphological and physiological identifi cations based on Gram staining, shape, motility, sporeformation, catalase, CO2 and NH3 production, and the ability to grow on temperature at 10oC and 45oC.Molecular identifi cation based on the amplifi cation of 16S rRNA gene. The potential application of selectedisolates for probiotics was evaluated based on the ability to grow on media with low pH and the additionof 0.5% bile salts, the ability to inhibit the growth of pathogenic Bacillus cereus and Eschericia coli, and in vitroadherence ability. On the basis of morphological, physiological and molecular analysis of 16S rRNA gene, itwas concluded that the selected isolate 1AF was a strain of Lactobacillus casei. Evaluation of probiotic in vitro showed that 60.4% of cells were resistant to pH 2.0 for 90 minutes. Survival of isolate 1AF after growing at0.5% bile salts was 70.8%. The selected isolate 1AF showed the ability to inhibit the growth of Eschericia coli and Bacillus cereus with inhibitory zone of 12.00±1,00 and 15.33±1.53 mm, respectively. In vitro study on theadherence value of isolate to solid plate was found at 46.5%. It is concluded that Lactobacillus casei isolate 1AFis a potential candidate as probiotics and subject to further in vivo evaluation
High Frequency Spontaneous Deletions within the IcaADBC Operon of Clinical Staphylococcus epidermidis Isolates.
No full textStaphylococcus epidermidis has been shown to undergo a phase variation correlating with expression of the icaADBC operon which contributes to biofilm formation. Biofilm formation of Enterococcus faecalis is related to heterogeneity in electrophoretic mobility. Here the relationship between phase variants of clinical isolates of S. epidermidis, icaADBC presence and electrophoretic mobility distributions is investigated. Of 105 S. epidermidis clinical isolates, 5 showed phase variation on Congo Red agar plate. Biofilm forming capability of the blackcolonies and inability of the red colonies were confirmed using a microtiter plate assay and confocal laser scanning microscopy. Upon analysis of electrophoretic mobility distributions, the black colonies displayed heterogeneity at pH 2 which was absent in the red colonies of the same strain. Surprisingly, it was shown that in all red colonies had lost the icaADBC genes. Determination of gene copy number using Real Time PCR targeting icaA showed reduction of gene copy within a culture with phase variation. In conclusion, using three fundamentally different approaches phase variation of the five clinical isolates was observed. Variants appeared through loss of icaA and icaC gens. To our knowledge this is the first report indicating S. epidermidis strains irreversible switching from biofilm + to biofilm – phenotype by deletion of ica genes. Key words: deletion, ica genes, Staphylococcus epidermidis, IcaADBC opero
Characterization of Haemolysin of Staphylococcus aureus Isolated from Food of Animal Origin
No full textStaphylococcus aureus is an important pathogen bacteria causing food poisoning and various infection in animals and humans. Haemolysin is one of the virulence factors of Staphylococcus aureus. The aims of the research were to characterize haemolysins of Staphylococcus aureus isolated from various food of animal origin, phenotypic- and genotypically. In the present study, eleven Staphylococcus aureus isolated from various food of animal origins from traditional markets and supermarkets in Yogyakarta, Sidoarjo, Jakarta, and Bandung were characterized for haemolysin, pheno- and genotypically. Characterization of haemolysin phenotypically based on haemolysis pattern of Staphylococcus aureus on sheep blood agar plate. Genes encoding hemolysin were amplified with specific primers by using polymerase chain reaction (PCR) technique. The results of the studies showed that Staphylococcus aureus on sheep blood agar plates revealed an alpha haemolysis pattern (18,18%), beta haemolysis (27,27%) and gamma haemolysis (54,55%). Based on amplification of the gene encoding haemolysin of Staphylococcus aureus with specific primers showed hla genes (81,81%), and hla combined with hlb genes (18,18%). The amplification of gene hla and hlb had a single amplicon with a size of approximately 534 bp and 833 bp, respectively. The haemolysin characteristics of Staphylococcus aureus from various food of animal origin could be used as important information to control staphylococcal food poisoning.Keywords : Staphylococcus aureus, haemolysin, PCR, food of animal origin
Determination of Haemaglutinin and Gene Encoding Fibronectin Binding Proteins Staphylococcus aureus Isolated from Dairy Milk Cows
No full textStaphylococcus aureus is a major pathogen causing clinical and subclinical mastitis in dairy milk cows. Themastitis has immense economical impacts, where it reduces of the quantity and quality of milk production.The aims of the research were to analyse haemaglutinin and gene encoding fbronectin binding proteins.Nineteen Staphylococcus aureus isolates used in the present study were isolated from dairy milk cows fromYogyakarta, Solo, Boyolali and Sumedang. The haemagluitinin of S. aureus were determined based onhaemaglutination reaction to erythrocytes of rabbit. Detection of gene encoding fbronectin binding proteinscould be performed with specifc primers using polymerase chain reaction (PCR). The results of studiesshowed that most of S. aureus (78,95%) expressed haemaglutinin based on their ability to aglutinate rabbiterythrocytes. Analysis of gene encoding fbronectin binding proteins of S. aureus revealed gene fnbA withsize of approximately 1300 bp for 57,89% isolates, gene fnbB with size of approximately 900 bp for 31,58%isolates and both of gene fnbA and fnbB could be detected for 31,58% isolates. The characters of S. aureusbased on haemaglutinin, gene fnbA and fnbB of the present study could be used as an information to controlof S. aureus infection in dairy herds
Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant
No full textMycobacterium 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
Biodegradation of Used Engine Oil by Acinetobacter junii TBC 1.2
No full textThe isolates have capability to degrade used engine oil was obtained from soil in the beach contaminatedwith used engine oil. One of the selected isolates TBC 1.2 was identified by its 16s rDNA as Acinetobacterjunii. The microorganism can use hydrocarbons in used engine oil as the sole carbon source and energy, alsoit significantly degraded almost all hydrocarbon compounds in used engine oil. With its ability Acinetobacterjunii TBC 1.2 has a potency to be utilized for bioremediation of soil polluted with used engine oil
Microorganisms Associated with Volatile Organic Compound Production in Spoilt Mango Fruits
No full textMicroorganisms associated with the production of volatile compound in spoilt mango fruits sold in Sokoto town were isolated and identified. The organisms include seven species of bacteria and a species of yeast. These include Bacillus pumilus, Bacillus firmus, Brevibacillus laterosporus, Morganella morganii, Paenibacillus alvei, Staphylococcus saccharolyticus, Listeria monocytogenes and Candida krusei respectively. GC-MS analysis revealed the presence of eleven and sixteen volatile organic compound in the healthy and spoilt ripe mango fruits. Octadecanoic acid, oleic acid, 1 – Butanol, 3 – methyl-, carbonate (2:1) and 3,7 – Dimethyl nonane were common to both healthy and spoilt fruits with the first three having higher concentration in healthy fruits than spoilt while the later had higher concentration in the spoilt. One methyl group of 3,3- Dimethyl hexane in healthy fruit was shifted to position two to yield 2,3-Dimethyl hexane in the spoilt fruits. 2,2-Dimethylbutane, Methyl(methyl-4-deoxy-2,3-di-O-methyl.beta.1-threo-hex-4-enopyranosid) urinate, 3-(4-amino-phenyl)-2-(toluene-4-sulfonylamino)-propionic acid, 2-Methyl-3-heptanone, 3,5-Nonadien-7-yn-2-ol, (E,E), Butanoic acid, 1,1-dimethylethyl ester, 1-methyl-3-beta.phenylethyl-2,4,5-trioxoimidazolidine, Pentanoic acid, 2,2-dimethyl, ethyl ester (Vinyl 2,2-dimethylpentanoate), 4-Methyurazole, 1-Tridecyn- 4 – 9 – ol, 1-Hexyl-1-nitrocyclohexane were unique to spoilt fruits. This study suggests that these unique volatile metabolites could be exploited as biomarkers to discriminate pathogens even when more than one disease is present thereby curbing post harvest loss during storage after further validation and the volatile organic compound could form the basis for constructing a metabolomics database for Nigeria