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All Journal INDONESIAN JOURNAL OF PHARMACY
M. Kuswandi
Faculty of Pharmacy Universitas Gadjah Mada Yogyakarta
Medicine & Pharmacology

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The antiviral activity of srikaya seed (Annona squamosa L.) ethanolic extract against newcastle disease virus in chicken embryo Nanik Sulistyani; Ibnatul Azizah; M. Kuswandi
Indonesian Journal of Pharmacy Vol 20 No 2, 2009
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (207.358 KB) | DOI: 10.14499/indonesianjpharm0iss0pp62-67

Abstract

Srikaya (A. squamosa L.) is a plant containing polyphenol, flavonoid, tannin, alkaloid and saponin compounds. These compounds could be extracted by ethanol solvent. Previous study shown that infuse of Srikaya seed had antiviral activity with value of IC50 3,236 'g/mL against Newcastle Disease Virus (NDV). Therefore, the aim of research is to know the potency of ethanolic extract of Srikaya seed as antiviral against Newcastle Disease Virus.The 28 chicken embryos at age of 9-11 days were classified into 7 groups, i.e. : control of virus, control of solvent ethanol : aquadest (1:10) and treatment groups were given ethanolic extract of Srikaya seed with concentration 0.1 'g/mL; 0.15 'g/mL; 0.2 'g/mL; 0.25 'g/mL; dan 0.3 'g/mL. NDV was inoculated into allantoic cavity intermediate after inoculated of extract and then incubated for 2 days. Antiviral activity were observed by hemaglutination method. The value of IC50 was calculated by probit analysis.The result showed that ethanol extract of srikaya seed (A. squamosa L.) had antiviral activity because of increasing concentration caused the higher inhibition percentage of hemaglutination. The value of IC50 is 0.152 'g/mL.Key words: Antiviral, Annona squamosa, Newcastle Disease Virus (NDV).
Sequencing analysis of the hole DNA of a mutant gene of Klebsiela pneumoniae resistant against BRL 41897A (KSL 19 Mutant) M. Kuswandi
Indonesian Journal of Pharmacy Vol 17 No 2, 2006
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (340.343 KB) | DOI: 10.14499/indonesianjpharm0iss0pp69-75

Abstract

Previous article reported that three positive transformants of different K.pneumonie resistant mutants against BRL 41897A (KSL 19, KSL38 and KSL62) carrying TnphoA was sequenced. The results showed that the three different transformants of the mutants had different genes inserted TnphoA. Subsequently, we then cloned the wild typeM10 chromosome fragments into pUC18 and recombinants analysed by gel electrophoresis and Southern blotting using HindIII fragment from KIH 19 as aprobe. The clones were designated M19 carrying the hole gene which had an important role in the antibiotic resistantion. The plasmid DNA of positive clone was isolated, analysed by Southern blotting, and sequenced using primers based on sequencing data from KIH19. Sequencing data of KIH19 confirmed the sequence of M19 and the point of insertion of transposon. The amino acid sequence at the N terminus of disrupted gene ksl19 showed the characteristics of features of signal peptide structure. In order to characterise the M19 clone, further complementation analysis was carried out, by transformation of M19 plasmid into KSL19 mutant cell, designated T19, and then checked by gel  electrophoresis and sensitivity test. The complementation studies showed that sensitivity to BRL 41897A was restored on transformation of a large (7 kb) fragment of DNA carrying ksl19 geneKey words: Resistant K.pneumoniae KSL19 mutants -BRL41897A, sequencing.
IDENTIFICATION OF Klebsiella pneumoniae RESISTANT MUTANTS AGAINST BRL 41897A ANTIBIOTIC USING SOUTHERN BLOT ANALYSIS. M. Kuswandi
Indonesian Journal of Pharmacy Vol 13 No 4, 2002
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (159.702 KB) | DOI: 10.14499/indonesianjpharm0iss0pp185-192

Abstract

Isolation of Klebsiella pneumoniae resistant mutants against BRL 41897A antibiotics using TnphoA has yielded 30 mutants which have been characterized. In order to identify whether the mutants are as a result of the insertion (transposition) of TnphoA, and to count how many copies of TnphoA are inserted in the each mutant chromosome, several chromosomal DNA of the mutants have been cut with restriction enzymes and analised using Southern blot method and DIG conjugation. The results showed that at least one mutant (KSL19) carried one copy of transposon, whereas at least 2 mutants (KSL38 and KSL52, ) have 2 copies. It is obvious that TnphoA mutagenesis provides an efficient method of selecting mutants which are defective in genes controlling expression of the outer membrane and the periplasmic membrane proteins.Key words: Resistant K. pneumoniae mutants- TnphoA mutagenesis – BRL 41897A- Southern Blot.
Co-Authors Ibnatul Azizah Nanik Sulistyani