Indonesian Journal of Biotechnology
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Diversity of Nonribosomal Peptide Synthetase Genes in the AnticancerProducing Actinomycetes Isolated from Marine Sediment in Indonesia
No full textMarine actinomycetes is a group of bacteria that is highly potential in producing novel bioactivecompound. It has unique characteristics and is different from other terrestrial ones. Extreme environmentalcondition is suspected to lead marine actinomycetes produce different types of bioactive compoundfound previously. The aim of this study was to explore the presence and diversity of NRPS genes in 14anticancer-producing actinomycetes isolated from marine sediment in Indonesia. PCR amplificationand restriction fragment analysis of NRPS genes with HaeIII from 14 marine actinomycetes were doneto assess the diversity of NRPS genes. Genome mining of one species of marine actinomycetes (strainGMY01) also was employed towards this goal. The result showed that NRPS gene sequence diversity in 14marine actinomycetes could be divided into 4 groups based on NRPS gene restriction patterns. Analysisof 16S rRNA gene sequences of representatives from each group showed that all isolates belong to genusof Streptomyces. Genome mining result showed that strain GMY01 harboring 10 different NRPS geneclusters that encode secondary metabolites, as pure NRPS or hybrid between NRPS and other compounds.These results indicated that marine actinomycetes having a high potential to be developed as source ofanticancer drugs development
Limited evidence for white spot syndrome virus susceptibility associated with expression of PmVRP15 in local population of giant tiger shrimp (Penaeus monodon)
No full textWhite spot syndrome virus (WSSV) is a devastating viral disease in shrimp aquaculture. Infection ofWSSV in penaeid shrimps affects immune defense and changes gene expression. PmVRP15 has been reported as a part of the WSSV propagation pathway that is highly up-regulated in hemocytes at the acute phase of WSSV infection. This study analyzed the expression of PmVRP15 in local populations of giant tiger shrimp (Penaeus monodon) to be associated with susceptibility to WSSV. Tested populations consisted of an inbreeding population (G8) and outbreeding population (G8iA) from Jepara, Indonesia. Susceptibility was determined by cumulative mortality, median lethal time (LT50), and severity of infection at time of death. Though all populations were susceptible to WSSV, the frst mortality in G8 occurred at 18 hours post-infection (hpi) with mild infection, while frst mortality of G8iA occurred at 30 hpi with severe infection. The LT50 of G8 was signifcantly lower than that of G8iA, indicating that G8iA was less susceptible to WSSV than G8. Relative PmVRP15 transcripts of G8iA were insignifcantly down-regulated, whereas relative PmVRP15 transcripts of G8were insignifcantly upregulated. Although it’s still not conclusive, the results of this study suggest that PmVRP15 has weak potentialas a WSSV susceptibility marker in G8 and G8iA broodstock selection
Synergistic effects of para-hydroxy meta-methoxy chalcone (pHmMC)- doxorubicin treatments on T47D breast cancer cells
No full textResistance to some cancer chemotherapeutic drugs has been identifed. One strategy to overcome that problem is by combining two or more of the drugs to get co-chemotherapeutic effects. A derivate chalcone, 3 - (4’-hydroxy-3’-methoxyphenyl)-1-phenyl-2-propene-1-on or para hydroxy meta methoxy chalcone (pHmMC), has been reported to have cytotoxic activity on some cancer cells through some pathways. The aim of this study was to investigate the effects of combinations of pHmMC and Doxorubicin (Dox) on the cytotoxicity, anti-proliferation, apoptosis, and the cell cycle of T47D (breast cancer cell-lines) in vitro. The cytotoxic and antiproliferative activity were determined by MTT (3-[4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide] assay. The combination index (CI) was used to determine the synergistic, additive or antagonistic effects of the combinations. Flowcytometry method was performed to determine the combination effects on the apoptosis and cell cycle. The results indicated that the combinations had a higher inhibitory effect on the cell growth compared to those of single treatments of pHmMC and Dox. All the doses used in the combinations were lower of the single doses at their IC50s. The results showed all the combinations gave synergistic (CI: 0.3 – 0.7) up to strong synergistic (CI: 0.1 – 0.3) effects. The synergistic effects of the combinations were due to increased apoptosis and induced cell cycle arrest in S and G2/M phases on the cancer cell lines
Detection and identifcation of adherence genes of intestinal-origin Lactobacillus and Pediococcus strains grown on gastric mucin in vitro
No full textOne of the primary selection criteria for potential probiotics is the ability to adhere to the host gastrointestinal tract. This study evaluated the in vitro adhesion ability on gastric mucin of two Lactobacillus casei strains (AP and AG) and two Pediococcus acidilactici strains (BE and BK), and identifed the corresponding genes responsible for adherence. Adhesion assays were performed in 96-well polystyrene microtiter plates using gastric mucin from porcine stomach as the matrix. An in vitro study on gastric mucin revealed that lactobacilli had a greater adherence ability compared with pediococci strains. The potential adherence genes were investigated using polymerase chain reaction (PCR) technology. Using specifc primers, PCR studies amplifed 150 base pairs of a potential mub gene and 161 base pairs of a potential ef-Tu gene, but no amplifed bands for potential map and bac genes were obtained. Sequence comparisons showed that the 150 and 161 amplifed base pairs were respectively homologous to the mub of Pediococcus acidilactici and ef-Tu genes of Lactobacillus paracasei. We concluded that the adherence ability of two strains of Lactobacillus casei (AP and AG) and two strains of Pediococcus acidilactici (BE and BK) on gastric mucin is in accordance with the presence of ef-Tu and mub genes. High level attachment in lactobacilli is likely to correlate with the ef-Tu gene, which is a lactobacilli-specifc adhesive gene
Isolation and caracterization of ficin enzyme from Ficus septica Burm F stem latex
No full textThis research aims to isolate and characterize the fcin enzyme from Ficus septica stem latex. Ficin from Ficus septica stem latex was isolated using column chromatography. Then enzyme activity was tested at different temperature (40oC, 50oC, 60oC, 70oC) and pH (6.0, 7.0, 8.0) levels. Ficin enzyme activity of joint treatment with variations in temperature and pH was analyzed using two-way ANOVA with a factorial pattern followed by Least Signifcant Difference (LSD) test. The results showed that temperature treatment signifcantly affects enzyme activity. However, the treatment of pH and the interaction between temperature and pH did not signifcantly affect the fcin enzyme activity. There was no signifcant difference in fcin activity at the incubation temperatures of 40oC and 50oC, as well as 60oC and 70oC. However, comparing the incubation temperatures of 40oC and 50oC with treatment 60°C and 70°C showed a signifcant difference in fcin enzyme activity. In the treatment of incubation at pH 6, 7 and 8 for fcin enzyme activity showed no signifcant difference. We concluded that the Ficus septica plant latex contained fcin enzyme with an optimum temperature of 60°C and optimum pH of 6, 7, and 8
Genetic Determination and Clonal Relationships of Staphylococcus aureus Isolated from Dairy Cows in Baturraden, Central Java, Indonesia
No full textCases of mastitis in cows at Baturraden are leading to signifi cant and ongoing problems due to reducedproduction and lower milk quality. This study was designed to identify which of selected virulence determinantgenes of S. aureus are involved in the Baturraden infection, and to determine the clonal relationship amongthese isolates. Seventeen isolates were identifi ed as S. aureus based on their biochemical properties and speciesspecifi city for 23S rRNA and nuc genes. S. aureus isolates were genotypically characterized for the selectedvirulence determinants: coa, clfA, fnbA, fnbB, cap5, spa IgG and spa X- region genes. Clonal relationship analysisamong isolates was carried out using AFLP and results compared with previously confi rmed relationshipsbetween selected S. aureus isolated from other regions. The results show that eight isolates contain all thegenes, but six isolates lack fnbB and two isolates lack cap5 genes. AFLP analysis showed that all isolates of S.aureus originating from cows in Baturraden belong to one cluster. This study provides additional knowledgeabout S. aureus infection in Baturraden cows, including the number of virulence determinant genes that mayplay a role in pathogenicity
Cytotoxicity of Buah Merah (Pandanus conoideus Lamk.) Extract on Breast Cancer Cell Line (T47D)
No full textBuah Merah (Pandanus conoideus Lamk.) has been extensively used to treat various diseases includingcancer. There are many varieties of buah merah and there was no scientifi c study comparing cytotoxicity ofdifferent varieties. The objective of this study was to investigate the cytotoxicity of three varieties of buah merahknown as Barugum, Maler and Yanggiru on breast cancer cell line (T47D). All samples were collected fromPapua, Indonesia. Each sample was extracted consecutively using three solvents chloroform, methanol andwater resulted to nine crude extracts. The cytotoxic activities were determined using MTT assay. The crudeextract showed the lowest IC50 was selected for further bioassay-guided fractionation. Fractionation was doneusing vacuum liquid chromatography coupled with preparative TLC to fi nd the active compounds. Severaldetection reagents were applied to TLC for identifi cation of the class of the potent compounds. The resultshowed that the potent extracts was obtained from Barugum methanol extract followed by Maler chloroformextract with IC50 value of 132.83 μg/ml and 139.72 μg/ml, respectively. All Yanggiru extracts did not showactivity. The bioassay-guided fractionation of Barugum and Maler extracts showed that the most potent fractioneluted by a mixture of hexane:ethyl acetate (75:25), was in Maler variety with IC50 value of 25,7 μg/ml, fourtimes higher than the most potent fraction of Barugum with IC50 value of 104,61 μg/ml. TLC analysis of themost potent fraction showed that the active compounds was class of terpene. Result of this study supportedthe utilization of buah merah Maler variety for breast cancer treatment
Allozyme variation of the endemic and vulnerable Dyera lowii Hook.f. in Central Kalimantan: Implications for genetic resources conservation
No full textDyera lowii is an endemic and vulnerable tree species of commercial value as chewing gum found inpeat swamp forests, scatteredly distributed in Sumatra, Kalimantan, and Peninsular Malaysia. Their existenceis now under severe threat due to habitat conversion. This study is aimed to assess genetic diversity withinfour natural populations (Hampangen, Parahangan, Sebangau, Selat Nusa ) and one plantation in CentralKalimantan based on allozyme variation. Electrophoresis procedures were conducted with an isoelectricfocusing polyacrylamide slab gel system. The result showed high genetic diversity (HE=0.52) and gene fl ow(3.402) seemed to be effective. A total of 14 alleles were found among all the analysed population. Meannumber of alleles per locus (Aa) was 3.206, and the effective number of alleles per locus (Ae) was 2.21. Geneticdifferentiation between populations (FST) was signifi cant at the moderately level (0.0685). Most allozymevariation was found within population (93.2%). Special attention is essential to conserve a private allele ofGot-1-e (9%) at Selat Nusa population. Sebangau population missed the alleles of Est-2-b and Got-1-a, as foundin other populations. Selat Nusa population is expected to enhance the effective management for geneticresources conservation of this vulnerable species in the future
The assessment of genetic variability and taxonomic affinity of local pummelo accessions from Yogyakarta, Indonesia based on RAPD
No full textPummelo (Citrus maxima) is one of three biotypes considered as true species within the genus Citrus.A major issue of pummelo taxonomy in Indonesia is the high number of cultivars showing variability inphenotypic characters but of unknown genetic diversity. In this study, the assessment of genetic variabilityand taxonomic affi nity of local accessions of C. maxima from Yogyakarta was examined based on RAPDfi ngerprinting. The availability of universal primers and technical simplicity makes RAPD as a molecular toolof choice for the assessment of genetic variability at various taxonomic levels. In this study, 13 accessions of C.maxima collected from Yogyakarta were observed for their genetic variability. An additional three registeredcultivars were included for comparative purpose. Two decamer primers used for the amplifi cation of DNAproduced 222 bands with 174 of them were polymorphic. The data was subjected to cluster analysis to observethe grouping of accessions and taxonomic affi nity. Results indicated high genetic variability among accessions.The dendrogram constructed using UPGMA method based on simple matching coeffi cient showed twomain clusters were which was in line to morphological characters. The grouping of accessions showed cleardifferentiation between accessions bearing white and those with reddish fruit fl esh, and thus demonstratestaxonomic value of this study in recognizing important agronomic character for this tropical fruit crop
Induced-Coagulated Plasma-Fibrin Gels as a Biological Scaffold for Cell Attachment and Proliferation of Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSC)
No full textFibrin gels are an ideal natural biological scaffold for tissue engineering because they are biocompatible,biodegradable, and have many biological surface markers. However, most research on fi brin gels used commercialfi brin kits that could be costly and limited in some areas. In this study, fi brin gels were made by inducing bloodcoagulation by adding a common diagnostic kit to assess the time for blood to clot, called activated partialthromboplastin time (aPTT). This induced coagulated plasma (iCoplas)-fi brin gels was evaluated for its ability toenhance biological activity of umbilical cord-derived mesenchymal stem cell (UC-MSC), which were cell attachmentand proliferation. Fibrinogen concentration had infl uence on cell attachment, where only 50% of the cells couldattach to 77 mg/dl fi brinogen gels whereas 93% cells adhered to 154 mg/dl fi brin gels. There were no signifi cantdifferences in cell proliferation on polysterene culture dish and fi brin gels (p>0.05). These results showed thatiCoplas-fi brin gels could be used as a fi brin-based scaffold, yielding no signifi cant difference than polysterene-tissueculture dish cultures in cell attachment and cell proliferation on 154 mg/dl fi brinogen concentration