Indonesian Journal of Biotechnology
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The Susceptibility of katG Ser315Thr Mycobacterium tuberculosis mutant Against Anti Tuberculosis Drugs
No full textMycobacterium tuberculosis resistant strains is a matter of great concern for TB control Program since nocure for some multi drug resistance TB (MDR-TB) cases. These strains could spread around the world, makea great challenges for control measures. Mycobacterium tuberculosis strains resistance to anti tuberculosis drugsis mainly caused by the alteration in several genes encoding the molecular targets. Mutation of katG at codon315 especially Ser315Thr are responsible for INH resistance in a large proportion of TB cases. The aim of thisstudy is to know the frequency of of katG Ser315Thr of M. tuberculosis mutant and the pattern of resistance toAnti TB drugs. The study design was observational laboratoric. Eighty fi ve M. tuberculosis INH resistant clinicalisolates were screened for mutation of katGSer315Thr by using PCR-RFLP and DNA sequence analysis.Theresults showed that katG Ser 315Thr were identifi ed in 23 (27,05%) of INH resistance isolates. Frequency ofMDR among the katG Ser315Thr mutants almost double than the non mutant. The result suggested that thethe katG Ser315Thr mutation may play important role in the development of MDR-TB
Impact of Curcuma mangga Val. Rhizome Essential Oil to p53, Bcl-2, H-Ras and Caspase-9 expression of Myeloma Cell Line
No full textCancer is a disease, a public health problem, which is found in the world as well as in Indonesia. Ingeneral, some of cancer theraphies are ineffective, characterized by the resistance performance of cancer cell line,the exposed normal cell and by the side effects. Nowadays, studies to fi nd the specifi c and safely anti-cancerdrugs were increased by the time. Several studies revealed that Curcuma mangga Val. Rhizome contains somesecondary metabolites, essential or non-essential oil, which has cytotoxic activities to the cancer cells. Basedon these anti-cancer potentials, this study has several aims to recognize anti-cancer selectivity and molecularmechanism by inducting apoptosis and inhibiting myeloma cell proliferation. To C. mangga Val. essential oil,immunocyto chemical test was performed to determine the expression of p53, caspase-9, Bcl-2, H-Ras proteinwhile TUNEL test was performed to determine the number of apoptosis cells.The results of this study shown that anti-cancer molecular mechanism of C. mangga Val. essential oil tomyeloma cell line was performed by increasing apoptosis; by increasing the expression of pro-apoptosis p53,caspase-9 protein and reducing protein which is increasing proliferation Bcl-2 and H-Ras
Effects of Light Quality on Vegetative Growth and Flower Initiation in Phalaenopsis
No full textThe effects of LEDs (Light-Emitting Diodes) emitting different colours namely red, blue, red andblue, and white lights on vegetative growth and fl ower initiation of Phalaenopsis have been evaluated.Phalaenopsis“otohine/taisuco fi re bird” seedlings in vitro were subjected to different light qualities for either2 or 4 weeks, and then each seedling was planted in a plastic pot containing sphagnum and grown in thegrowth chamber under similar light quality for 3 months. For fl ower induction, mature Phalaenopsis plantshaving 4 – 6 leaves were grown for 3 months in the growth chamber under different light qualities. The leafspan, chlorophyll, gibberellin and cytokinin content were determined. In addition, the expressions of FT-likegene in the leaf, axillary bud, fl ower bud and stalk were examined.Vegetative growth was enhanced under blue, red-blue or white LEDs compared to that of the control.Gibberellin and cytokinin content increased in the seedlings subjected to white LEDs. Based on the averageof leaf span increment it was suggested that the growth of Phalaenopsis seedlings can be promoted by givingeither blue, red-blue or white LEDs. From the second experiment, it was found that fl ower induction inPhalaenopsis can be obtained in plants that had just fi nished fl owering without the application of LEDs. Theexpression of FT-like gene in the leaf as well as fl ower bud and stalk suggests that this gene is involved infl ower regulation of Phalaenopsis
Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii
No full textPromoter regulates expression level of foreign gene in transgenic organism. This study was performed to select asuitable promoter as the fi rst step towards production of valuable trait-enhanced seaweed by transgenic technology. Greenfl uorescent protein (GFP) gene was used as a reporter to determine the activity of promoter in seaweed Kappaphycusalvarezii. GFP gene constructs driven by cytomegalovirus (pCMV-GFP), caulifl ower mosaic virus (pCaMV-GFP),medaka β-actin (pmBA-GFP) and Japanese fl ounder keratin (pJfKer-GFP) promoters were introduced by electroporationmethod. Electroporation was performed using a gene pulser (BIORAD) with voltage of 300 V, pulse length of 0.5 ms,pulse numbers of 4, and pulse interval of 0.1 s. Promoter activity was determined by analyzing GFP gene expressionlevel using a fl uorescent microscope. The results showed that CMV regulated highest number of fi lament callus(34.10%±1.49) expressing GFP at medium to strong fl uorescence levels. CaMV promoter had relatively similar activitywith CMV, but lower number of fi lament callus expressing GFP (10.48%±0.25). mBA promoter drove GFP expressionat medium level and similar number of fi lament callus (8.85%±2.31) expressing GFP with CaMV, while JfKer promoterhad lowest activity by means in number of fi lament callus expressing GFP (4.79%±0.26) and GFP expression level. PCRanalysis for transgenic confi rmation showed a DNA band of PCR product from pCMV-GFP and pCaMV-GFP expressingfi lament callus in the same size (about 0.6 kb) with positive control of plasmid. Thus, CMV and CaMV promoters wasan appropriate promoter and foreign gene could be transferred to fi lament callus by electroporation method. Combiningthis achievement with developing a culture method of fi lament callus to be thallus, stable transgenic breeding in K.alvarezii can be feasible
In Vitro Screening of Falcataria moluccana (Miq.) with Gall Rust (Uromycladium tepperianum (Sacc.) Filtrate as Media Selection
No full textIn vitro screening of Falcataria moluccana (Miq.) was conducted by tissue culture method. Seeds fromtwo different site of community forest, 400 m (S1) and 800 m (S2) above sea level, were used as material.Double concentration of MS (Murashige & Skoog, 1962) with 40 mg/l gall rust (Uromycladium tepperianum(Sacc.) fi ltrate were used for media selection. The results of this research showed that 66 % axenic plantlets invitro from S1 and 27 % from S2 were still survived after 3 months incubation without subculture. The meanof fresh weight (2. 21 ± 0. 26 g) and dry weight (1. 97 ± 0. 12 g) from S1 plantlets lower than the mean of freshweight (2. 87 ± 0,18 g) and dry weight (2. 16 ± 0. 14 g) from S2 plantlets. Qualitative of terpenes, saponins andquantitative of total phenolics were analyzed from those gall rust extract, as source of fi ltrate media, attackedand un-attacked of F. moluccana. They all qualitatively have capability to produce terpenoid and saponin. Itis notice that U. tepperianum, un-cultured pathogen, contain of those compound that may play a role as codeterminantsof pathogenecity. While the highest total phenolic compound were contained in gall rust extract(2. 35 %), followed by attacked F. moluccana branches (1. 18 %) and un-attacked F. moluccana branches (0. 44%). This indicated that phenolic compound in gall rust has higher activity as a response of F. moluccana to U.tepperianum pathogen pressures and result of this study suggest the great value of gall rust fi ltrate for use asmedia selection in vitro
Histological Features of Catecholaminergic Neuron in Substantia Nigra Induced by Paraquat Dichloride (1,1-dimethyl-4,4 bipyridinium) in Wistar Rat as A Model of Parkinson Disease
No full textParaquat dichloride has been used by farmers as a herbicide to kill the grass. On the other hand, paraquatdichloride is harmful if enters to the body, causing Parkinson’s disease, since it is disrupting dopamineproduction in the substantia nigra pars compacta or dopamine pathways Nigro striatal pathway. The studywas done to fi nd out the histological changes of catecholaminergic neurons and Nigro striatal pathway causedby paraquat dichloride treatment in Wistar rats as a model of Parkinson’s disease.Twenty-two Wistar rats 3,5 months old were divided into 4 groups, 5 rats each. Group I (control group)were injected with aquabidest, while groups II, III, and IV were injected intraperitoneally with paraquatdichloride in aquabidest, with the dosage 5 , 10 and 15 mg/kg bw respectively. The rats were injected onceper week for 6 weeks. Three days after the last injection, the rats were anesthetized using xylasin (2 mg/kg)and ketamine (20 mg/kg) intramuscularly, and then were intracardiac perfused using physiological saline asprerinse solution, followed by 10% buffered formalin solution as a fi xative. After animals were fi xed, the brainswere removed and embedded in paraffi n block and cut in 12 μm thickness for immunohistochemistry stainingusing tyrosine hydroxylase antibody. The results of staining then were observed under light microscope andanalyzed descriptively.The results showed that the catecholaminergic neurons were distributed in the substantia nigrapars compacta in all treatment groups, however, the cell density were found decreased only in group IV.Catecholaminergic neurons appear in the bipolar and multipolar form, while dopamine ‘Nigro striatal pathway’was found exist in all treatment groups. From our study, histologycally the decreased of catecholaminergicneurons is only found in rats that received paraquat dichloride in dose 15 mg/kg bw for 6 weeks
The synthesis of polymeric dual-functional antimicrobial surface based on poly(2-methyl-2-oxazoline)
No full textThere is a high interest in the development of antimicrobial coatings to fi ght bacterial infections.We present the development of dual-functional antimicrobial surface, in which a biopassive platform wasfunctionalized with bioactive compounds on the surface, using a graft copolymer system poly(L-lysine)-graftpoly(2-methyl-2-oxazoline)-quarternery ammonium compound (PLL-g-PMOXA-QAC). Alkyne functionalitywas introduced to the PMOXA chain at α-terminus by initiating the living cationic polymerization of 2-methyl-2-oxazoline with a propargylic-initiator. The reaction was terminated with carboxy derivative-terminator thatallows grafting of the polymeric chain from the β-terminus to poly(L-lysine) (PLL) backbone, resulting in graftcopolymer alkynyl PLL-g-PMOXA. The conjugation between alkynyl PLL-g-PMOXA and QAC was thenperformed using click reaction. The chemical structures of the polymers were characterized by MALDI-TOFspectrometry and NMR spectroscopy. The results demonstrate that we have successfully synthesized PLL-g-PMOXA-QAC copolymer with grafting density (number of lysine/number of PMOXA) of 0.33. The resultingPLL-g-PMOXA-QAC copolymer was then immobilized onto carboxylated tissue cultured polystyrene (TCPS)surface and exposed to bacteria solution to test its dual-functional properties. Preliminary live-and-deadbacteria study indicates dual-functionality of the PLL-g-PMOXA-QAC-coated surface
Expression analysis of antioxidant genes in response to drought stress in the fl ag leaf of two Indonesian rice cultivars
No full textThe objective of this study was to analysis the expression of antioxidant genes in response to drought stress in Indonesian rice. The malondialdehyde (MDA) content and the expression of Cu-ZnSod1, cCu-ZnSod2, MnSod1, cApxa, cApxb, chl-sApx, Cat1, Cat2, Cat3, Gr1, Gr2, and Gr3 genes were assayed in the rice fl ag leaf of Ciherang and Situ Bagendit cultivars subjected to control, mild and severe drought during the grain filling phase. Increase in MDA content of Ciherang treated to mild and severe drought was almost two-fold and three-fold respectively, while MDA content in Situ Bagendit subjected to mild and severe drought increased approximately one-fold and two-fold as compared to the control. The semi quantitative reverse transcription polymerase chain reaction (sqRT-PCR) analysis showed that the expression of cCu-ZnSod1, MnSod1, Cat2, Gr3 genes of Ciherang, and cCu-ZnSod2, MnSod1, cApxa, cApxb, chl-sAPX, Cat2 and Gr1 genes of Situ Bagendit increased in flag leaf of plant treated to drought. Expressions of cApxb, chl-sApx, Cat3 of Ciherang and Cu-ZnSod1 and Gr2 genes of Situ Bagendit were not changed signifi cantly by drought stress. Decreased expression was shown by cCu-ZnSod2, cApxa, Cat1, Gr1 and Gr2 genes of Ciherang, and Cat1, Cat3 and Gr3 genes of Situ Bagendit. The results indicated that the activity of oxidative defense was regulated by four genes; cCu-ZnSod1, MnSod1, Cat2, Gr3 in Ciherang, and eight genes; cCu-ZnSod1, cCu-ZnSod2, MnSod1, cApxa, cApxb, chl-sApx, Cat2 and Gr1 in Situ Bagendit. Therefore, differences in the number of antioxidant genes controlling oxidative defense system might determine the difference of the oxidative defense capacity between both cultivars in response to drought stress during grain fi lling
Phylogenetic relationship of Gram Negative Bacteria of Enterobacteriaceae Family in the Positive Widal Blood Cultures based on 16S rRNA Gene Sequences
No full textThe purpose of this study was to analyze the phylogenetic relationship of Gram negative bacteria (3strains of Salmonella typhi, 1 strain of Escherichia coli, 1 strain of Serratia marcescens, and 3 strains of Enterobactercloacae) of Enterobacteriaceae family in positive Widal blood cultures based on 16S rRNA gene sequences. Theresults respectively showed that each two 16S rRNA gene clones of Serratia marcescens KD 08.4 had a closerelationship with 16S rRNA gene of Serrratia marcescens ATCC 13880 (similarity: 99.53-99.8%), Eschericia coliBA 30.1 with Eschericia coli ATCC 11775T (similarity: 99.38-99.67%), Salmonella typhi BA 07.4, Salmonella typhiKD 30.4, and Salmonella typhi SA 02.2 with Salmonella typhi ATCC 19430T (similarity: 99.4-100%) as well as theisolates of Enterobacter cloacae SA 02.1, Enterobacter cloacae BA 45.4.1, one 16S rRNA gene clone of Enterobactercloacae TG 03.5 with Enterobacter cloacae ATCC 23373 (similarity: 99.0-99.87%)
Poly-β-Hydroxybutyrate (PHB) Production By Amylolytic Micrococcus sp. PG1 Isolated From Soil Polluted Arrowroot Starch Waste
No full textPoly-β-hydroxybutyrate (PHB) production from amylolytic Micrococcus sp. PG1. Poly-β-hydroxybutyrate(PHB) is an organic polymer, which synthesized by many bacteria and serves as internal energy. PHB ispotential as future bioplastic but its price is very expensive due to glucose usage in PHB industry. Thedevelopment of PHB production using starch as an alternative carbon source has been conducted to reducethe dependence of glucose in PHB production. In this study, amylolytic bacteria from arrowroot processingsite were screened quantitavely based on amylase specifi c activity and PHB producing ability. The result of thestudy showed that among of 24 amylolytic isolates, 12 isolates of them were able to accumulate PHB rangedfrom 0,68-11,65% (g PHB/g cdw). The highest PHB production from substrate arrowroot starch was PG1 andafter optimization resulted in increasing of PHB production up to 16,8% (g PHB/g cdw) 40 hours incubationtime. Based on morphological, biochemical and physiological characters, the PG1 isolate was identifi ed asMicrococcus sp. PG1. Result of the FTIR analysis of produced polymer by Micrococcus sp. PG1 was indicatedas poly-β- hydroxybutyrate (PHB