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Cloning of mutant genes of Klebsiellae pneumoniae resistant against BRL 41897A (KSL Mutants)
Get PDFPrevious observations to identify Klebsiellae pneumoniae mutants (KSL) resistant to BRL41897A antibiotic (Kuswandi, 2002), using Southern blot dan SDS-PAGE analysis, showed that there were variation of TnphoA copy in each mutant and differentiation of outer membrane (OM and IROMP) of several mutants cell wall. In order to clone the gene which carries TnphoA from the KSL mutants, the chromosomal DNA fragments that had been ligated to pUC18 plasmid was transformed into E.coli host cells. The positive transformants of different mutants (KSL 19, KSL38 and KSL62) carrying TnphoA has been sequenced. The results showed that the three different transformants of the mutants had different genes inserted TnphoA.Key words: Resistant K.pneumoniae mutants -BRL41897A, clone, sequencing
Cytotoxicity testing of alkaloid compounds isolated from sponge Petrosia sp: its potency for development of anticancer agent
Get PDFCancer is still a major problem and common cause of death around the world. Various therapeutic agents have been developed to fight against cancer, but none of these agents give satisfactory results and without debilitating side effects. A number of researches have been conducted to search anticancer compounds with renewed vigour.Sponges, marine invertebrates, are known as rich sources of compounds which pronounced pharmacological activities. The aims of this study are to determine cytotoxic effect of two toxic compounds isolated from chloroform fraction of Petrosia sp sponges collected from Bunaken on myeloma cells.The two toxic compounds were isolated based on bioassay guidedisolation on brine shrimp larvae. Isolation was conducted using column chromatography followed by preparative TLC. Cytotoxic effect of the two compounds was conducted in 96 well plate using RPMI 1640 as medium. The number of viable cells was determined using MTT assay and LC50 (μg/mL) of the compounds was analysed using probit analysis.The results showed that the two compounds were alkaloid and toxic to larva A. salina with LC50 of 7.23 (compound 1) and 5.69 μg/mL (compound 2). These compounds were also toxic to myeloma cells with LC50 values of 16.95 μg/mL (compound 1) and 18.8 μg/mL (compound 2). The longer the incubation time, the compounds were more toxic as showed by the lower LC50 values .Key words: cytotoxic, Petrosia sp, Artemia salina Leach, myelom
DUAL EFFECTS OF FLAVONOID QUERCETIN ON RAT BASOPHILIC LEUKEMIA (RBL-2H3) CELLS : INHIBITS HISTAMINE RELEASE AND REDUCES CELL GROWTH
Get PDFThe dual effects of the flavonoid quercetin on rat basophilic leukemia cells (RBL-2H3), tumor analog mast cells, was studied. Quercetin is known as an anti-inflammatory drug that inhibits mast cell secretion of histamine. This study aims to investigate the consequences of varying incubation times with quercetin on 2H3 cells to histamine release inhibitory action and the effects of long time incubation to the morphology of the cells. The effect of quercetin on histamine synthesis was also observed. The histamine release from the cells was inhibited by quercetin as expected, but the ability of secretion was rapidly recovered when quercetin was removed before challenging. Incubation up to 6 hours decreased the inhibitory action, but longer than 6 hours increased the inhibitory activity. In long time incubation, the cells exhibited cell damage, decreased cell growth, morphological changes, and detachment from the underlying surface in proportion with the concentration of quercetin. The instant and reversible inhibitory effects of quercetin appear to represent a first phase of actions, while reduced cell growth, elevated cell damage and morphological changes seem to be connected to a second phase. Consequently, quercetin could be considered as a compound that acts dually on RBL-2H3 cells.Key words : quercetin, histamine, RBL-2H3 cell
The bioavailability of furosemide-polyethylene glycol (PEG 4000) solid dispersion in male rabbits
Get PDFFurosemide is a potential diuretic drug usually used for the secondary treatment of hypertension. Unfortunately this agent is very slightly soluble in water, so it has poor bioavailability. The oral administration of drug shows that only 60 % of the dose can be absorbed.The aim of the present study is to increase the bioavailability of furosemide by mixing the furosemide-PEG 4000 solid dispersion (1 : 1) in capsule dosage forms (formula A). The powder of LasixR tablet in capsules was used as a standard reference (formula B), and the original powder of furosemide in capsules was used as control (formula C). All of the formulas contained 40 mg of furosemide.The bioavailability of these formulas was evaluated following oral administration in male rabbits using The Latin Square Cross Over Design, then the plasma furosemide concentrations were analyzed spectrofluorometrically. The results indicated that the bioavailability of furosemide in the formula A was equal to that of the formula B (P > 0,05) or both formulas were bioequivalent. But the Cpmax value of the formula C was significantly lower in comparison with the formulas A and B.Key words: furosemide, solid dispersion, bioavailability
CLEANING EFFECTIVITY OF SEVERAL SURFACTANS TO PESTISIDES RESIDUES ON FRESH APEL FRUITS
Get PDFIntensification efforts in farming to increase productivity must consider the pesticide utilization, especially insecticide and herbicide. Several pestisides which are still used include carbofuran and organochlorine, some of them have lipofilic properties and might harm to human health. Therefore, an effort is required to washing off pesticides from farming products is one of the effort which can be performed. Since pesticides has lipofilic properties, therefore cleaning pesticides with water is not sufficient. Surfactant is required to increase washing off ability of water. Wash off ability of several surfactants circulated on the market i. e. SL, ML and A were investigated. The result showed that the wash off ability values of surfactants to DDT residues on fresh apples were 79.18 %, 75.19 % and 67.49 % for SL, A and ML respectively. The wash off effectiveness of surfactant A, SL and ML to -metrine were 85.29 %, 80.48 % and 64.47 % respectively.Key words: pesticide, cleaning efficiency surfactant, DDT, -metrine
Jaspamide: Structure identification of cytotoxic and fungicide compound isolated from Stylissa flabelliformis sponges
Get PDFA research on the structure identification of cytotoxic and fungicide compound of Stylissa flabelliformis sponge has been conducted. The structure identification was analysed with spectroscopy ultraviolet, MS, 1HNMR and 13C-NMR methods.Based on the spectroscopic data and comparison with literatures, the compound was identified as jaspamide.Key words: sponge, Stylissa flabelliformis , jaspamide, structure elucidatio
EFFECT OF RIFAMPICIN PRETREATMENT ON HIPOGLYCEMIC EFFECT OF GLYPIZIDE AMONG HEALTHY VOLUNTEERS
Get PDFThe incidence of tuberculosis in diabetic patients is high, therefore, the combination usage of antituberculosis (rifampicin) and antidiabetic (glypizide) medicines is inevitable. Rifampicin, an enzyme inductor, capable to influence the metabolism of other medicine when administered in concordance. The aimed of this study, therefore, was to investigate the influence of rifampicin pre-treatment on hypoglycaemic effect of glypizide (a second generation of sulphonylurea) among 12 Indonesia healthy volunteers, of both sexes. This study applied randomized crossover design receiving with and without rifampicin pre-treatment. Prior to starting the experiment, the pre-treatment group was given 450 mg of rifampicin orally daily for 7 days. Subsequently, single dose of 5 mg glypizide was administered to both of control and pre-treatment groups. The blood samples were then collected at a certain interval time for 7 hours. Glucose Oxydase (GOD) method were used to analyze the level of glucose in blood samples. The result showed no significant influence of rifampicin on hypoglycaemic effect of glypizide. However, it was found that rifampicin reduced significantly blood glucose level at 2.5 and 3 hours for 51.38 % and 20.58 %, respectively. The conclution of this study exhibit that pre-treatment with 450 mg of rifampicin daily for 7 days did not affect AUC0-7 blood glucose level as a result of single dose administration of 5 mg glypizide.Key words : rifampicin, glypizide, hypoglycaemic effec
Solubilization capacity of surfactant due to its different chain length of lipophile and hydrophile
Get PDFThe aim of the study is to look at the influence of different chain length of hydrophile and lipophile of the surfactants to their solubilization capacity. The solubilization capacity of surfactants with different chain lengths of lipophile and hydrophile has been performed by preparing somesolubilization form formulas, using several kinds of lipophilic chain length, i.e. cetyl (C-16), stearyl (C-18), and oleyl (C=18 with unsaturated chain) alcohols and ethyleneoxyde chain length of 2, 10 and 20 respectively. After preparation, the solubilited forms were then stored at temperature of 25O and 45OC, and observed if there were any solubilization occurred. The results have showed that it needs an optimum length of ethyleneoxyde to have better solubilization; at certain longer ethylenoxyde the longer the length of lipophilic chain, the more solubilization would be, therefore, unsaturated hydrocarbon chain caused diminution of solubilization; whatever the surfactant used as an agent of solubilization, storage at a higher room temperature would facilitate the solubilization to occur.Key words : surfactant, solubilizatio
IDENTIFICATION OF IROMP PROTEIN OF K.PNEUMONIAE RESISTANT MUTANTS AGAINST BRL41897 USING SDS –PAGE
Get PDFInitial characterization, the MIC, Fe uptake and detection of siderophores, of the resistance K.pneumoniae mutants against BRL41897A (KSL mutants) showed that there were differences amongs them. Therefore it was necessary to observe the OMP and IROMP proteins of the mutants especially which had different MIC’s to the WT galur (M10). Results from SDS-PAGE analysis showed that KSL19 produced 49kDa protein weakly and similar result was found in the 22 kDa protein production by KSL38, KSL52, KSL58 and KSL59. Using Fe-CAA media -media lack of Fe - showed that 3 mutants synthesized certain protein weaker than M10 galur, KSL19 in the production of the 88 kDa protein, KSL38 and KSL59 at the 80 kDa protein. We also observed that KSL19 synthesized new 88 kDa protein. This result showed that certain mutant which had decreased production of one protein could stimulate another weak protein.Key words: SDS-PAGE – IROMP – K.pneumoniae mutants – BRL 41897A.
The stability of PGV-0 (Pentagamavunon-0) as an Antiinflammatory drug in liquid dosage forms
Get PDFPGV-0 is a curcumin derivate, a synthetic compound which may be a candidate of a new drug. This substance has a potent inflammatory effect with a very low toxicity.One of the first step which must be searched for a candidate of drug, is to perform a stability study. There are many degraded drugs causing the adverse reactions. Most of them could be the initiator in forming an antigen at anaphylactic reaction or allergic reaction. Further more some of the degraded drugs are very toxic. So, the study of degradation or the stability investigations of a new drug should be carried out.The PGV-0 stability in a buffer solution at pH 10,0 had been studied by an accelerated temperature method. The temperatures were held at 50°, 55° and 60° C. The intact PGV-0 has been analysed by HPLC. The results then were used to define the shelf-life, the half life and the activation energy of the degradation of PGV-0.It was evident that PGV-0 was unstable in aqueous solution, the shelf-life was only 45.3 hours, the half-life was 299 hours and the activation energy was 14,2 kkal mol-1. Because the PGV-0 was not stable in aqueous solution, it is suggested that this substance should be made into solid dosage forms instead of the liquid dosage forms.Key word : PGV-0, stability, solutio