In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3 U/ml. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF 923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13 U/ml) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCo_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567 U/mg. The molecular weight of PLD was about 55 kD and the optimum pH and temperature were 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$. Special metal ions were not necessary to the PLD activity.
Purpose: Analysis of volatile organic solvents in 2-deoxy-2-$[^{18}F]$ fluoro-D-glucose ($[^{18}F]$FDG) preparations was performed by gas chromatography (GC), in accordance with USP. Materials and Methods: Analyses were carried out on a Hewlett-Packard 6890 gas chromatography equipped with an FID. Results: We determined the amounts of ethanol and acetonitrile on every batch of our routine $[^{18}F]$FDG preparations, ranging between 5000 ppm and 100 ppm. In our routine preparation of $[^{18}F]$FDG, the amount of acetonitrile and ethanol in the final product were well below the maximum allowable limit described in the USP. Conclusion: Our $[^{18}F]$FDG preparations were in accordance with the suggested USP maximum allowable levels of the quality control analysis of volatile organic compounds.
We isolated wild lactic acid bacteria from kimchi and identified as Lactobacillus brevis by using API 50 CHL Kit, some morphological and physiological tests. In order to evaluate the acid tolerance of Lactobacillus brevis, its survivals rate, glycolysis assay, membrane permeability, and pH profiles of $H^+-ATPase$ were also determined. When Lactobacillus brevis were incubated in Lactobacilli MRS broth adjusted to various levels of pH for 2 hours, the decreases in its population at pH 4.0 and 3.0 were about 2.61 log cycles/mL and 5.89 log cycles/mL, respectively, but there was no decrease at pH 6.0 and 5.0. Glycolysis by Lactobacillus brevis had optimal pH about 6.5 and glucose degradation was reduced by 50% at a pH of 5.2. $Mg^{++}$ release from Lactobacillus brevis cells in medium with pHs of 4.0 and 3.0 was 24.3 and 71.2% of totals, respectively. The $H^+-ATPase$ of Lactobacillus brevis showed a maximal activity between pH values of approximately 6.5 to 7.0.
Statement of problem: Streptococcus produces energy and forms extracellular polysaccharides by metabolizing sucrose. Insoluble glucan, a kind of extracellular polysaccharide, is the important material of dental plaque. Fructose affects the metabolism of sucrose. Purpose: The purpose of this study was to evaluate the effect of fructose on the metabolism of sucrose in Streptococcus mutans. Materials and methods: To determine the effect of fructose on the formation of artificial plaque by Streptococcus mutans Ingbritt, S. mutans and fructose were placed in beakers containing M17 broth and sucrose. The wires were hung on frameworks inserted into cork stoppers, and then immersed in each of the beakers. After the incubation with gentle shaking, each wire was weighed. To analyze the effect of fructose on the sucrose metabolism by S. mutans or glucosyltransferase, S. mutans and fructose were placed in M17 broth containing sucrose. After the incubation. the remaining sucrose and polymers were analysed by thin layer chromatography. Results: The following results were obtained; 1. When Streptococcus mutans was cultured in the media containing 3% sucrose for 8 hours, the mean weight of formed artificial plaque on the wires was $124.3{\pm}3.0mg$, whereas being reduced to $20.7{\pm}10.2mg$ in the media added with 3% sucrose and 4% fructose(p<0.05) 2. When the control containing glucose was added with sucrose, the optical density of Streptococcus mutans solution cultured for 24 hours was not increased compared with the control, while being increased by adding with fructose. 3. When Streptococcus mutans was incubated in the media added with sucrose and fructose for 8 hours, the number of viable cells was increased compared with the media added with sucrose. 4. The amount of remained sucrose was increased in Streptococcus mutans culture supernatant of media added with sucrose and fructose than with sucrose only. but the amount of produced insoluble glucan was decreased. 5. The amounts of remained sucrose and produced soluble glucan were increased in the culture of glucosyltransferase-contained media added with sucrose and fructose than with sucrose only, but the amount of produced insoluble glucan was decreased . Conclusion: These results indicated that the sucrose metabolism and the production of insoluble glucan were inhibited in Streptococcus mutans by adding fructose in the media containing sucrose.
This research was concentrated to Investigate the effects of Insumon as a newly developed dietary supplement in patients with type 2 diabetes. The average glycated hemoglobin level was dropped from 7.0${\pm}$1.1% to 6.7${\pm}$0.9% after 8 weeks with statistic significance (p=0.0202). For patients of HbA1c over 7%, the average HbA1c significantly decreased from 8.1${\pm}$0.8% to 7.5${\pm}$0.8% (p=0.0171). The average fasting blood glucose level was decreased significantly from 9.5${\pm}$1.2 mmol/L (171.6${\pm}$22.3 mg/dL) to 8.5${\pm}$1.5 mmol/L (152.3${\pm}$26.3 mg/dL) (p=0.0262) after 4 weeks and to 8.8${\pm}$1.5 mmol/L (158.3${\pm}$24.1 mg/dL) (p=0.0445) after 8 weeks in the group of patients of FBG of over 7.8 mmol/L (140 mg/dL). The average HDL cholesterol level was increased significantly after 8 weeks from 1.2${\pm}$0.2 mmol/L (46.5${\pm}$8.6 mg/dL) to 1.3${\pm}$0.3 mmol/L (51.6${\pm}$10.9 mg/dL) (p=0.0007). Thus, these results suggested that patients with type 2 diabetes might benefit by Insumon as a herbal dietary supplement without any serious side effects.
Objective : Recently scolopendrid aqua-acupuncture has been a good effect on pain control but it has not been known about clinical safety. The purpose of this study was to investigate acute toxicity of scolopendrid aqua-acupuncture. Method : In order to prove the clinical safety of scolopendrid aqua-acupuncture, We have observed a bacteriological examination and clinical pathology test after scolopendrid aqua-acupuncture treatment. Balb/c mice were injected intravenous with Scolopendrid aquaacupuncture treatment for $LD_{50}$ and acute toxicity test. We analyzed physical reaction(side effect)and clinical pathology test before and after Scolopendrid aqua-acupuncture treatment of mice and 20 patients suffering from pain, who admitted department of Acupunture and Moxibustion, College of Oriental Medicine, Won-Kwang University Kwangju hospital. Results : In the Blood agar plate and Nutrient agar plate, a bacteriological examination did not show a bacillus. In acute $LD_{50}$ toxicity test, there was no mortality thus unable to attain the value. Examining the toxic response in the acute toxicity test, there was no sign of toxication. In acute toxic test, running biochemical serum test couldn't yield any differences between the control and experiment groups. In the 20 patients treated with Scolopendrid aqua-acupuncture, hematologic test did not show remarkable change. In the 20 patients treated with Scolopendrid aqua-acupuncture, Liver function test(AST, ALT, ALP) showed a slight decrease on the contrary, and abnormal rate showed a decrease of 5.0% compared with previous study. Reanl function test(BUN, Cr) and abnormal rate showed a decrease of 5.0% compared with previous study. In the 20 patients treated with Scolopendrid aqua-acupuncture, Electrolyte were normal range before and after treatment. In the Urine analysis of 20 patients, Leukocyte, Protein, Glucose, Keton, Bilirubin, U-bilinogen were not detected before and after Scolopendrid aqua-acupuncture treatment, and the rest almost made no difference.
Gopal, Velmani;AL Rashid, Mohammad Harun;Majumder, Sayani;Maiti, Partha Pratim;Mandal, Subhash C
Journal of Pharmacopuncture
/
v.18
no.2
/
pp.7-18
/
2015
Objectives: Lawsone (1,4 naphthoquinone) is a non redox cycling compound that can be catalyzed by DT diaphorase (DTD) into 1,2,4-trihydroxynaphthalene (THN), which can generate reactive oxygen species by auto oxidation. The purpose of this study was to evaluate the toxicity of the phytomarker 1,4 naphthoquinone and its metabolite THN by using the molecular docking program AutoDock 4. Methods: The 3D structure of ligands such as hydrogen peroxide ($H_2O_2$), nitric oxide synthase (NOS), catalase (CAT), glutathione (GSH), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH) and nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) were drawn using hyperchem drawing tools and minimizing the energy of all pdb files with the help of hyperchem by $MM^+$ followed by a semi-empirical (PM3) method. The docking process was studied with ligand molecules to identify suitable dockings at protein binding sites through annealing and genetic simulation algorithms. The program auto dock tools (ADT) was released as an extension suite to the python molecular viewer used to prepare proteins and ligands. Grids centered on active sites were obtained with spacings of $54{\times}55{\times}56$, and a grid spacing of 0.503 was calculated. Comparisons of Global and Local Search Methods in Drug Docking were adopted to determine parameters; a maximum number of 250,000 energy evaluations, a maximum number of generations of 27,000, and mutation and crossover rates of 0.02 and 0.8 were used. The number of docking runs was set to 10. Results: Lawsone and THN can be considered to efficiently bind with NOS, CAT, GSH, GR, G6PDH and NADPH, which has been confirmed through hydrogen bond affinity with the respective amino acids. Conclusion: Naphthoquinone derivatives of lawsone, which can be metabolized into THN by a catalyst DTD, were examined. Lawsone and THN were found to be identically potent molecules for their affinities for selected proteins.
The contribution of endogenous transport systems to the blood-brain barrier (BBB) transport of basic and acidic drugs was studied by using a carotid injection technique in rats and an isolated bovine cerebrovascular disease state were compared between the normotensive rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) which have been well established as an animal model with pathogenic similarities to humans. Basic drugs such as eperisone, thiamine and scopolamine inhibited, in a concentration dependent manner the in vivo uptake of $[{^3}H]choline$ through BBB, whereas amino acids and acidic drugs such as salicylic acid and valproic acid did not inhibit the uptake. The uptake of $[^3H]choline$ by B-CAP increased with time and showed a remarkable temperature dependency. The uptake of $[^3H]choline$ by B-CAP showed the very similar inhibitory effects as observed in the in vivo brain uptake, and was competitively inhibited by a basic drug, eperisone. The in vivo BBB uptakes of $[^3H]acetic$ acid and $[^{14}C]salicylic$ acid were dependent on pH of the injectate and the concentration of drugs. Several acidic drugs such such as salicylic acid, benzoic acid and valproic acid inhibited the in vivo uptake of $[^3H]acetic$ acid, whereas amino acid, choline and a basic drug such as eperisone did not inhibit the uptake. The uptake of acetic acid by B-CAP was competitively inhibited by salicylic acid. The permeability surface area product (PS) through BBB for $[^3H]choline$ in SHRSP was significantly lower than that in WKY. The concentration of choline in the brain dialysate in SHRSP was about half of that in WKY, while no significant difference was observed in the plasma concentration of choline between SHRSP and WKY. No significant difference was observed in the transport of monocarboxylic acids, glucose and neutral amino acid through BBB between SHRSP and WKY. From these results, it was concluded that BBB transport system of choline contributes to the transport of basic drugs through BBB, that acidic drugs can be transported via a moncarboxylic acid BBB transport system and that the specific dysfuntion of the BBB choline transport in SHRSP was ascribed to the reduction of the maximum velocity of choline concentration in the brain interstitial fluids.
The effect of green tea drinking on the hepatocellular chemical cacinogenesis have been studied. Placental glutathione S-transferase(GST-P) positive foci area in a liver tissue, contents of thiobarbituric acid reactive substances(TBARS), total cytochrome P450 and glucose 6-phospphatase(G6P) activity in hepatic microsomes were investigated. Weaning Sprague-Dawley male rats were fed AIN-76A diet with deionized water or green tea infusion, Rats of CTR and CTR+ groups were provided deionized water while GTI and GTI+ groups were provided green tea instead of deionized water for the entire experimental period of 13weeks. Rats of GTP and GTP + groups had deionized water for the first 6 weeks and switched to green tea for the last 7weeks of the experimental period. CTR+, GTI +, and GTP + groups were carcinogen treated groups, Diethylnitrosamine(DEN) was injected as a single dose of 200mg/kg body weight intraperitoneally after 4 weeks of feeding. 2-Acetyla-minofluorene(AAF) was used as a carcinogen proliferater and suppled in the diets of carcinogen treated rats as 0.02% content for the last 6weeks starting from 2weeks after DEN injection. Rats were sacrificed after 13week weeks of feeding. The area and number of GST-P positive foci detected in carcinogen treated rats were decreased by green tea ingestion but when timing and duration of green tea ingestion was delayed after promotion period as in GTP + group, GST-P positive foci were not decreased as much as in GTI+ group. TBARS contents of carcinogen treated rats decreased by 13weeks of green tea ingestion but GTP groups did not show statiscally significant differences. G6P activities tended to decrease by carcinogen treatment but changes were not statiscally significant by green tea ingestion. Total cytochrome P450 contents were increased by carcinogen treatment. Thirteen weeks of green tea ingestion (GTI) also increased to total cytochrome P450 contents while 7weeks of green tea ingestion(GTP) did show any effects. These results suggest that green tea has suppressive effects on hepatocellular chemical carcinogenesis probably through the activities of antioxidant compounds. (Korean J Community utrition 2(5) : 735∼744, 1997)
This study was conducted to measure the difference in the total antioxidant status(TAS) of overweight and normal weight adults, and to investigate the correlation between TAS and the nutrient intake. Anthropometric parameter, TAS, biochemical parameters and dietary intake were measured in the normal weight group(n=17, BMI 20-25, obesity index 90-110%, body fat 16-20%) and overweight group(n=13, BMI > 25, obesity index > 120%, body fat > 25%) among Korean young males. The TAS of the overweight group was significantly higher than that of the normal weight group(p < 0.01). Among biochemical parameters, the average uric acid concentration of the overweight group was significantly higher(p < 0.05) compared to that of the normal weight group indicating, the uric acid concentration reflects TAS. Usual dietary intake showed that the intakes of Ca and vitamin B$_2$, are below RDA. The average intake of vitamin C in the overweight group was significantly lower than that in the normal group(p < 0.05). There was no difference in the intake of carotene and vitamin E between groups. The diet quality of both groups was satisfactory. The result of the correlation analysis on TAS, the uric acid concentration, and the measured indexs are as follows : In the overweight group, TAS was negatively correlated with alchol drinking, and uric acid concentration was positively correlated with BMI, obesity index, and TAS. In conclusion, TAS was significantly higher in the overweight group than that of the normal group, although no association was found with nutrient intakes. Serum uric acid showed a positive relationship with TAS indicating, it is a possible determinant of the antioxidant capacity.
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