• Journal of Soil and Groundwater Environment
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• v.21 no.5
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• pp.16-24
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• 2016

A new strain of Pseudomonas sp. was isolated from mercury (Hg)-contaminated sites in Taiwan. This bacterium removed more than 80% of Hg present in the culture medium at 12 h incubation and was chosen for further analysis of the molecular mechanisms of Hg tolerance/removal abilities in this Pseudomonas sp. We used RNA-seq, one of the next-generation sequencing methods, to investigate the transcriptomic responses of the Pseudomonas sp. exposed to 60 mg/L of Hg²⁺. We de novo assembled 4,963 contigs, of which 10,533 up-regulated genes and 5,451 down-regulated genes were found to be regulated by Hg. The 40 genes most altered in expression levels were associated with tolerance to Hg stress and metabolism. Functional analysis showed that some Hg-tolerant genes were related to the mer operon, sulfate uptake and assimilation, the enzymatic antioxidant system, the HSP gene family, chaperones, and metal transporters. The transcriptome were analyzed further with Gene Ontology (GO) and Cluster of Orthologous Groups (COGs) of proteins and showed diverse biological functions and metabolic pathways under Hg stress.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.98-98
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• 1997

This study was done to investigate the effect of vitamin E on hypoxia/reoxygenation-induced hepatic injury in isolated perfused rat liver. Rats were pretreated with vitamin E or vehicle(soybean oil). Isolated livers from fasted 18 hours were subjected to 45min of low flow hypoxia or N$_2$ hypoxia followed by reoxygenation for 30min. The perfusion medium used was KHBB(pH 7.4) and 50${\mu}$㏖/$\ell$ of ethoxycoumarin was added to the perfusate to determine the ability of hepatic drug-metabolizing systems, In low flow hypoxia model, total glutathione and oxidised glutathione levels were significantly increased by hepoxia/reoxygenation with slight increase in LDH levels. These increases were prevented by vitamin E pretreatment. In N$_2$ hypoxia model, LDH, total glutathione and oxidized glutathione levels were increased significantly by hypoxia but restored to normal level by reoxygenation. Vitamin E had little effect on this hypoxic damage. There were no significant changes in the rate of hepatic oxidation of 7-EC to 7-HC in both hepoxic models. But, the subsequent conjugation of 7-HC by sulfate or glucuronic acid were significantly decreased by hypoxia, but restored by reoxygenation in both hypoxia models. As opposed to our expectation, treatment with vitamin E aggrevated the decrease of the rate of conjugation and even inhibited the restoration by reoxygenation. Our findings suggest that hypoxia/reoxygenation diminishes phase II drug metabolizing function and this is, in part, related to decreased energy level.

• Korean Journal of Pharmacognosy
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• v.33 no.4 s.131
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• pp.312-318
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• 2002

Effect of the acid hydrolysate of silk fibroin on obesity was investigated in obese(057BL/KsJ-db/db) mice. After 8 weeks feeding of 1%(w/w) or 3%(w/w) fibroin hydrolysate, the extents of reduction in body weight were significantly higher than that of obese control. The weight reduction in female mice was higher than that in male mice. Plasma leptin in male mice increased up to 1.8-fold higher level than obese control by feeding hydrolysate. In case of female mice, however, it rather decreased with increased feeding concentration of hydrolysate. From the results of high glycine and serine contents of peptide fractions contained in fibroin hydrolysate, it was inferred that fibroin peptides might affect xylosyltransferase(XT) activity on chondroitin sulfate synthesis causing to change susceptibility of adipocytes to hormones such as insulin followed by the reduced leptin synthesis in female mice. The result of the higher lipolysis in hydrolysate-fed group than obese control indicated that the reduction in body weight was due to the increased lipolytic activities in male and female mice in common.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1067-1075
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• 2007

Twenty eight 3-4 month old castrated Black Bengal kids (Capra hircus) were used to determine the effects of source and level of dietary copper (Cu) concentration on their performance and nutrient utilization. Cu was supplemented (0, 10, 20 and 30 mg/kg diet DM) as copper sulfate ($CuSO_4$, $5H_2O$) or copper proteinate (Cu-P). Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each) and mineral and vitamin mixture (0.5% each) supplements at 3.5% of body weight to meet NRC (1981) requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. $CuSO_4$ or Cu-P was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well ventilated shed with facilities for individual feeding in aluminum plated metabolic cages. Blood samples were collected from the jugular vein on d 0, 30, 60 and 90 to determine hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and serum enzymes (alkaline phosphatase, alanine transferase and aspertate transferase). A metabolism trial of 6 days duration was conducted after 90 days of experimental feeding. Statistical analysis revealed that source and level of Cu supplementation improved live weight gain (p<0.04) and average daily gain (p<0.01). No significant contribution of source and level of Cu to alter serum serum enzymes was evident. Goats fed Cu-P tended to have higher Hb, PCV and TEC than with $CuSO_4$ supplementation. Cu-P increased digestibility of ether extract (EE, p<0.02) and crude fiber (p<0.05) and showed an increasing trend (p<0.09) for digested crude protein (CP) and crude fiber (CF). Supplemental dose of Cu linearly improved (p<0.02) digestibilities of dry matter (DM), organic matter (OM), EE and nitrogen free extract (NFE). Though the absorption of nitrogen (N) was not affected (p>0.10) by both source and dose of Cu, N retention was affected (p<0.04) and there was a significant $Source{\times}Dose$ interaction (p<0.05). Final body weight (BW) was not influenced (p>0.10) by the source of Cu but increasing dose of Cu increased (p<0.04) the BW of kids. TDN intake (g/kg $W^{0.75}$) was higher (p<0.05) with the increased dose of Cu and there was a significant $Source{\times}Dose$ interaction. It was concluded that supplementation of Cu from different sources and varying dose level in a concentrate based diet may improve performance, nutrient utilization and plane of nutrition in castrated Black Bengal kids. The effects on performance and nutrient utilization are more pronounced with Cu-P than $CuSO_4$ supplementation. Higher dose of Cu showed better result than lower dose.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.8
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• pp.1154-1161
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• 2000

Two experiments were conducted to evaluate the efficacy of cupric citrate (Cu-citrate) relative to cupric sulfate $(CuSO_4)$ as a Cu source for weanling and grow-finish pigs. In addition, the use of liver and bile Cu concentrations as indices of the bioavailability of Cu sources was investigated. Experiment one consisted of a nursery phase (35 d; initial BW=6.4 kg, final BW=21.4 kg) followed by a grow-finish phase (103 d; initial BW=21.5 kg, final BW=111.7 kg). Experiment two only consisted of a nursery phase (35 d; initial BW=6.3 kg, final BW=18.6 kg). Dietary treatments were identical for both experiments and consisted of: control (10 ppm $CuSO_4$); control+66 or 225 ppm $CuSO_4$; control+33, 66, or 100 ppm Cu-citrate. An antibiotic was included in diets for Exp. 1 but not Exp. 2. In both experiments, growth performance variables were similar for pigs receiving Cu-citrate and $CuSO_4$; however, growth performance was not improved by high concentrations of $CuSO_4$. Liver and bile Cu were increased (p<0.05) by 225 ppm $CuSO_4$; however, lower dietary concentrations of Cu from either $CuSO_4$ or Cu-citrate did not affect the Cu concentration of liver or bile relative to that observed in the control pigs. Irrespective of Cu source, there was no linear (p>0.10) increase in plasma Cu with increasing Cu concentrations in the diet for both experiments. However, the plasma Cu concentrations were highest (p<0.10) in pigs receiving diets supplemented with 225 ppm $CuSO_4$. Sixteen randomly chosen pigs per treatment in Exp. 1 were continued through the grow-finish phase. Body weight gain and feed intake were improved (p<0.10) by 66 ppm $CuSO_4$, but other dietary Cu treatments did not alter pig performance compared to the control diet. Plasma Cu concentrations were increased (p<0.10) by 225 ppm $CuSO_4$ in the growing phase and by 225 ppm $CuSO_4$ and 100 ppm Cu-citrate in the finishing phase. These data reveal no consistent effect of $CuSO_4$ on performance; therefore, it is difficult to assess the efficacy of these two Cu sources. In addition, these studies demonstrate that liver and bile Cu are not good indicators of Cu bioavailability in pigs fed adequate to pharmacological concentrations of Cu.

• Journal of Ginseng Research
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• v.11 no.2
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• pp.145-252
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• 1987

Isolated rat adipocytes are well known to possess opposite pathways of lipid metabolism: lipolysis and ipogenesis. Both of the metabolism respond to various biologically active substances such as epinephrine, ACTH and insulin. Epinephrine and ACTH stimulate lipolysis and insulin accelerates lipogenesis. Recently, Korean red ginseng powder was found to contain adenosine and an acidic poptide which inhibited epinephrine-induced lipolysis and sl imulated insulin-mediated lipogenesis from added glucose. The acidic peptide is consisted mainly of glutamic acid and glucose. Ginsenosides Rb1 and Re inhibited ACTH-induced lipolysis in isolated rat adipocytes, while they did not affect insulinstimulated lipogenesis, Thus, all these substances extracted from Korean red ginseng exhibited selective modulations toward the opposite metabolic pathways in rat adipocyte; They inhibited the lipolysis but not the lipogenesis. We call these substances"selective modulators". Recently, we isolated a toxic substance named "toxohormone-L " from ascites fluid of patients with various malignant tumors. The toxohormone-L stimulated lipolysis in rat adipocytes and induced anorexia in rats. Both the lipolytic and the anorexigenic actions of toxohormone-L were found to be inhibited by ginsenoside Rb2 in Korean red ginseng. Based on these results, physiological signifi¬cances of these substances in Korean red ginseng were discussed. Pan ax ginseng is a medicinal plant long used in treatment of various pathological states including general complaints such as head ache, shoulder ache, chilly constitution and anorexia in cancer patients, There have been many pharmacological studies on Panax ginseng roots. Petkovllreported that oral administration of an aqueous alcoholic extract of ginseng roots decreased the blood sugar levtl of rabbits. Saito2lreported that Panax ginseng suppressed hyperglycemia induced by epinephrine and high carbohydrate diets. These findings suggest that Panax ginseng roots contain insulin-like substances. Previously, we demonstrated that gin¬seng roots contain an insulin-like peptide which inhibits epinephrine-induced lipolysis and stimulated insulin-mediated lipogenesis. In 1984, we suggested that such an insulin-like substance should be called a selective modulator4). Present investigation describes the details of the selective modulators in ginseng roots. During progressive weight loss in patients with various neoplastic disease, depletion of fat stores have been observed. The depletion of body fat during growth of neoplasms is associated with increase in plasma free fatty acids. Recently, we found that the ascites fluid from patients with hepatoma or ovarian tumor and the pleural fluid from patients with malignant lymphoma elicited fatty acid release in slices of rat adipose tissue in vitro. The lipolytic factor, named"toxohormone-L". was purifed from the ascites fluid of patients with hepatoma. The isolated preparation gave a single band on both disc gel electrophoresis and sodium dodecyl sulfate(SDS)-acrylamide gel electrophoresis in the presence of ${\beta}$-mercaptoethanol. Its molecular weight was determined to be 70,000-75,000 and 65,000 by SDS-acrylamide gel electrophoresis and analytical ultracentrifugation, respectively. Injection of toxohormone-L into the lateral ventricle of rats significantly suppressed food and water intakes. There was at least 5 hr delay between its injection and appearance of its suppressive effect. In the present study, we also tried to find a inhibitory substance toward toxohormone-L from root powder of ginseng.

• Applied Biological Chemistry
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• v.22 no.3
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• pp.145-149
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• 1979

The radiochemical purity of $^{14}C(U)-glucose$ solution to be injected to normal laying hen was investigated for studying biokinetics of carbohydrate and lipid metabolism. The liquid scintillation counter was employed for determining the activity of carbon-14. The barium hydroxide and zinc sulfate were adopted to precipitate the protein in the solution. The glucose content in the solution was observed as 0.912 mg per ml, applying Hultman's method. The specific activity of $^{14}C(U)-glucose$ solution was known as 31.3 nCi/mg glucose. The glucose pentaacetate was synthesized to isolate the pure glucose from the solution. The specific activity of pure glucose was measured as 28.5 nCi/mg glucose. Therefore, it was known that the radiochemical purity of the solution was 82.7%.

• Journal of Microbiology and Biotechnology
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• v.31 no.10
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• pp.1409-1419
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• 2021

A growing number of healthy dietary ingredients in fruits and vegetables have been shown to exhibit diverse biological activities. Phloretin, a dihydrochalcone flavonoid that is abundant in apples and pears, has anti-inflammatory effects on ulcerative colitis (UC) mice. The gut microbiota and metabolism are closely related to each other due to the existence of the food-gut axis in the human colon. To investigate the interplay of faecal metabolites and the microbiota in UC mice after phloretin treatment, phloretin (60 mg/kg) was administered by gavage to ameliorate dextran sulfate sodium (DSS)-induced UC in mice. Gut microbes and faecal metabolite profiles were detected by high-throughput sequencing and liquid chromatography mass spectrometry (LC-MS) analysis, respectively. The correlations between gut microbes and their metabolites were evaluated by Spearman correlation coefficients. The results indicated that phloretin reshaped the disturbed faecal metabolite profile in UC mice and improved the metabolic pathways by balancing the composition of faecal metabolites such as norepinephrine, mesalazine, tyrosine, 5-acetyl-2,4-dimethyloxazole, and 6-acetyl-2,3-dihydro-2-(hydroxymethyl)-4(1H)-pyridinone. Correlation analysis identified the relations between the gut microbes and their metabolites. Proteus was negatively related to many faecal metabolites, such as norepinephrine, L-tyrosine, laccarin, dopamine glucuronide, and 5-acetyl-2,4-dimethyloxazole. The abundance of unidentified Bacteriodales_S24-7_group was positively related to ecgonine, 15-KETE and 6-acetyl-2,3-dihydro-2-(hydroxymethyl)-4(1H)-pyridinone. The abundance of Christensenellaceae_R-7_group was negatively related to the levels of 15-KETE and netilmicin. Stenotrophomonas and 15-KETE were negatively related, while Intestinimonas and alanyl-serine were positively related. In conclusion, phloretin treatment had positive impacts on faecal metabolites in UC mice, and the changes in faecal metabolites were closely related to the gut microbiota.

• Journal of Life Science
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• v.18 no.8
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• pp.1036-1041
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• 2008

Methenyltetrahydrofolate synthetase extract was obtained from mouse liver and purified via $30{\sim}70%$ ammonium sulfate fractionation, Fast Q anion exchange and phenyl agarose chromatography. HPLC gel chromatography and SDS-polyacrylamide electrophoresis experiments showed that the enzyme is a monomer with molecular weight of 23 kDa. Optimum temperature and pH were $35^{\circ}C$ and 6.5, respectively. The enzyme was chemically modified only by tetranitromethane and 1-ethyl-3- (3-dimethyl aminopropyl)-carbodiimide (EDC), indicating that tyrosine and carboxylate are in the active site. pH studies showed that 2 tyrosines are involved in the binding of the substrates and a carboxylate in catalysis. Therefore, the chemical mechanism of the enzyme is likely that 2 tyrosines bind to ATP and 5-formylTHFand a carboxylate acts as a general base.

• Proceedings of the Korean Society of Food Science and Nutrition Conference
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• 2004.11a
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• pp.65-70
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• 2004

Modulation of biotransformation enzymes is one mechanism by which a diet high in fruits and vegetable may influence cancer risk. Inhibition of cytochrome P450s (CYP) and concomitant induction of conjugating enzymes are hypothesized to reduce the impact of carcinogens in humans. Thus, exposure to types and amounts of phytochemicals may influence disease risk. Like other xenobiotics, many classes of phytochemicals are rapodly conjugated with glutathione, glucuronide, and sulfate moieties and excreted in urine and bile. In humans, circulating phytochemical levels very widely among individuals even in response to controlled dietary interventions. Polymorphisms in biotransformation enzymes, such as the glutathione S-transferases (GST), UDP-glucuronosyltransferases (UGT), and sulfotransferases (SULT), may ocntribute to the variability in phytochemical clearance and efficacy; polymorphic enzymes with lower enzyme activity prolong the half-lives of phytochmicals in vivo. Isothiocyanates (ITC) in cruciferous vegetables are catalyzed by the four major human GSTs: however reaction velocities of the enzymes differ greatly. In some observational studies of cancer, polymorphisms in the GSTMI and GSTTI genes that result in complete lack of GSTM1-1 protein, respectively, confer greater protection from cruciferous vegetable in individuals with these genotypes. Similarly, we have shown in a controlled dietary trial that levels of GST-alpha-induced by ITC-are higher in GSTMI-null individuals exposed to cruciferous vegetablse. The selectivity of glucuronosyl conjugation of flavonoids is dependent both on flavonoid structure as well as on the UGI isozyme involved in its conjuagtion. The effects of UGI polymorphisms on flavonoid clearnace have not been examind; but polymorphisms affect glucuronidation of several drugs. Given the strong interest in the chemopreventive effects of flavonoids, systematic evaluation of these polymorphic UGTs and flavonoid pharmacokinetics are warranted. Overall, these studies suggest that for phytochemicals that are metabolized by, and affect activity of, biotransformation enzymes, interactions between genetic polymorphisms in the enzymes and intake of the compounds should be considered in studies of cancer risk. Genetic polymorphisms in biotransformation enzymes may account in prat for individual variation in metabolism of a wide range of phytochemicals and their ultimate impact on health.