• Journal of Preventive Medicine and Public Health
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• 제35권4호
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• pp.282-286
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• 2002

Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

• Journal of Pharmaceutical Investigation
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• 제25권3호spc1호
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• pp.7-20
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• 1995

Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$ $Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.

• Microbiology and Biotechnology Letters
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• 제34권2호
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• pp.166-173
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• 2006

This research has been performed to clarify the relationship between hydrogeochemistry and bacterial community structure in groundwater contaminated with landfill leachate. We collected and analyzed samples from 5 sites such as leachate (KSG1-12), treated leachate (KSG1-16), two contaminated groundwaters (KSG1-07 and KSG1-08) and non-contaminated groundwater (KSG1-13). pH was 8.83, 8.04, 6.87, 6.87 and 6.53 in order; redox potential (Eh) 108, 202, 47, 200 and 154 mV; electric conductivity (EC) 3710, 894, 1223, 559 and 169.9 $\mu$S/cm; suspended solids (SS) 86.45, 13.74, 4.18, 0.24 and 11.91 mg/L. In KSG01-12, the ion concentrations were higher especially in $Cl^-$ and $HCO_3^-$ than other sites. The concentrations of Fe, Mn and $SO_4^{2-}$ were higher In KSG1-07 than in KSG1-08, and vise versa in $NO_3^{2-}$. In the comparison of DGGE fingerprint patterns, the similarity was highest between KSG1-13 and KSG1-16 (57.2%), probably due to common properties like low or none contaminant concentrations. Otherwise KSG1-08 showed lowest similarities with KSG1-13 (25.8%) and KSG1-12 (27.6%), maybe because of the degree of contamination. The most dominant bacterial species in each site were involved in $\alpha$-Proteobacteria (55.6%) in KSG1-12, $\gamma$-Proteobacteria (50.0%) in KSG1-16, $\beta$-Proteobacteria (66.7%) in KSG1-07, $\gamma$-Proteobacteria (54.5%) in KSG1-08 and $\beta$-Proteobacteria (36.4%) in KSG1-13. These results indicate that the microbial community structure might be changed according to the flow of leachate in grounderwater, implying changes in concentrations of pollutants, available electron accepters and/or other environmental conditions.

• KOREAN JOURNAL OF CROP SCIENCE
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• 제53권2호
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• pp.171-178
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• 2008

In the methanol extract from safflower seeds, two kinds of antioxidant were detected by preparative HPLC [$\mu$-Bondapak $C_{18}$ column ($7.8{\times}300\;mm$)]. Two unknown compounds were defined as CA and CB which had peaks at 22.1 min and 24.5 min, respectively. Antioxidant activity was measured by their scavenging ability on the stable tree radical of 1,1-diphenly-2-picrylhydrazyl (DPPH). For bulk extraction of antioxidants, the methanol extract was fractionated with hexan, chloroform, ethyl acetate and butanol. The ethyl acetate traction showing the highest DPPH radical scavenging activity was further purified by silica gel column chromatography to CA and CB. By NMR analysis, CA and CB were identified as N-(p-Coumaroyl)serotonin and N-feruoylserotonin, respectively. The content of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were analyzed by reverse phase HPLC using a $\mu$-Bondapak $C_{18}$ column ($3.9{\times}300\;mm$) with linear gradient elution from 10% acetonitrile to 50% acetonitrile for 30min on UV detector at 300 nm. The contents of N-(p-Coumaroyl)serotonin and N-feruoylserotonin were 4.11 mg/g DW and 7.29 mg/g DW, respectively, and these two DPPH radical scavengers were detected only in the hull of seeds.

• The Korean Journal of Physiology and Pharmacology
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• 제2권2호
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• pp.251-260
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• 1998

Cromakalim (BRL 34915), known as an airway smooth muscle relaxant, inhibited the releases of mediators in the antigen-induced mast cell activation. It has been suggested that cromakalim, in part, inhibited mediator releases by inhibiting the initial increase of 1,2-diacylglycerol (DAG) produced by the activation of the other phospholipase system which is different from phosphatidylcholine-phospholipase D pathway. The aim of this study is to further examine the inhibitory mechanism of cromakalim on the mediator release in the mast cell activation. Guinea pig lung mast cells were purified by using enzyme digestion and percoll density gradient. In purified mast cells prelabeled with $[^3H]PIP_2$, phospholipase C (PLC) activity was assessed by the production of $[^3H]$insitol phosphates. Protein kinase C (PKC) activity was assessed by measuring the protein phosphorylated from mast cells prelabeled with $[{\gamma}-32P]ATP$, and Phospholipase $A_2\;(PLA_2)$ activity by measuring the lyso-phosphatidylcholine produced from mast cell prelabeled with 1-palmitoyl-2-arachidonyl $phosphatidyl-[^{14}C]choline$. Histamine was assayed by fluorometric analyzer, and leukotrienes by radioimmunoassay. The PLC activity was increased by activation of the passively sensitized mast cells. This increased PLC activity was decreased by cromakalim pretreatment. The PKC activity increased by the activation of the passively sensitized mast cells was decreased by calphostin C, staurosporine and cromakalim, respectively. The $PLA_2$ activity was increased in the activated mast cells. The pretreatment of cromakalim did not significantly decrease $PLA_2$ activity. These data show that cromakalim inhibits histamine release by continuously inhibiting signal transduction processes which is mediated via PLC pathway during mast cell activation, but that cromakalim does not affect $PLA_2$ activity related to leukotriene release.

• Ocean and Polar Research
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• 제39권1호
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• pp.23-34
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• 2017

This study was carried out to investigate the spatial distributions of the macrobenthic communities in the Seomjin River estuary from May, 2015 to May, 2016. The number of species was 163, the mean density was $1,865ind.m^{-2}$, biomass was $204g{\cdot}wet\;m^{-2}$ during this study period. The highest number of species and density appeared among polychaetes whereas the most biomass was contributed by mollusks due to the presence of Corbicula japonica in every season. The study area was divided into 3 regions with similar benthic fauna responding to the gradient of the salinity. Praxillella praetermissa was the dominant species in regions of over 30 psu during all seasons. C. japonica and Hediste diadroma were dominant in the upper regions of the Seomjin River where the salinity was less than 10 psu. Heteromastus filiformis showed the broadest distributional range and dominated in all seasons except for the most upper stream at st. 7. From the result of the Bio-Env analysis, salinity was the most important environmental factor affecting the formation of macrobenthic communities in the study area, and salinity and TOC were the highest contributors to the macrobenthic communities. From the correlation analysis between major dominant species and environmental factors, C. japonica, Prionospio japonica and H. diadroma showed a negative correlation with salinity, while P. praetermissa and Scolectoma longifolia showed a positive correlation. H. filiformis was little affected by salinity but showed a positive correlation with TOC or silt content of sediment.

• Journal of Microbiology and Biotechnology
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• 제23권11호
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• pp.1617-1626
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• 2013

The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

• Journal of Embryo Transfer
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• 제25권4호
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• Environmental Engineering Research
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• 제11권3호
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• pp.127-133
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• 2006

Aerobic floc-like sludge was formed in a batch reactor and the effect of cations on the formation of aerobic floc-like sludge was studied. In order to enhance the formation (rate) of aerobic floc-like sludge, cations such as $K^+$, $Na^+$, $Ca^{2+}$, and $Mg^{2+}$ were added to the seed sludge. It was found that $Ca^{2+}$ had positive effect on the formation of floc-like sludge, as measured by sludge volume index (SVI) for settle ability. The formation of floc-like sludge was confirmed by the microscopic observation after DAPI staining. The scattered forms of sludge samples at the initial stage became aggregated to form floes after $Ca^{2+}$ addition. To ensure the functions of sludge floes in a treatment plant, the gradient of ionic species around the surfaces of floc-like sludge was monitored by ion selective microelectrodes for ${NH_4}^+,\;{NO_3}^-$, and pH. The effective concentration of $Ca^{2+}$ ion to form floc-like sludge was determined to be 750 mg/L (0.15 mg $Ca^{2+}/mg$ MLSS). Under the effective $Ca^{2+}$ condition, the SVI value was the lowest and large distribution of nitrifying bacteria at the outer surface was observed in the aerobic floc-like sludge. From the results, it was found that the calcium ion functioned as an agent for the formation of aerobic floc-like sludge, resulting in the enhanced nitrification.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.226-233
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• 2017

Jeung-pyun, a fermented rice cake, is prepared by fermenting rice sourdough using makgeolli, a traditional Korean rice wine, in the presence of yeast and lactic acid bacteria (LAB). The goal of this study was to conduct biochemical and microbial analyses of five different rice sourdoughs, each fermented with a different commercial makgeolli, using culture-dependent and culture-independent approaches. All sourdough samples fermented with different makgeolli for 6.5 h showed different profiles in pH, total titratable acidity, organic acid concentration, and microbial growth. LAB belonging to different genera were identified based on colony morphology on modified MRS and sourdough bacteria agar medium. PCR-denaturing gradient gel electrophoresis analyses of the five sourdoughs showed different bands corresponding to LAB and yeast. 16S/26S rRNA gene sequence analyses of the samples confirmed that the predominant LAB in the five fermented rice doughs was Lactobacillus plantarum, Lb. pentosus, and Lb. brevis. Various other Lactobacillus spp. and Saccharomyces cerevisiae were common in all five fermented samples. This study provides comprehensive and comparative information on the microflora involved in fermentation of rice sourdough and signifies the need to develop effective starters to enrich the quality of jeung-pyun.