Glucuronidation by the uridine diphosphateglucuronosy-ltransferase 1A enzymes (UGT1As) is a major pathway for elimination of particular drugs and endogenous substances, such as bilirubin. We examined the relation of eight single nucleotide polymorphisms (SNPs) and haplotypes of the UGT1A gene with their clinical factors. For association analysis, we genotyped the variants by direct sequencing analysis and polymerase chain reaction (PCR) in 218 healthy Koreans. The frequency of UGT1A1 polymorphisms, -3279T>G, -3156G>A, -53 $(TA)_{6>7}$, 211G>A, and 686C>A, was 0.26, 0.12, 0.08, 0.15, and 0.01, respectively. The frequency of -118 $(T)_{9>10}$ of UGT1A9 was 0.62, which was significantly higher than that in Caucasians (0.39). Neither the -2152C>T nor the -275T>A polymorphism was observed in Koreans or other Asians in comparison with Caucasians. The -3156G>A and -53 $(TA)_{6>7}$ polymorphisms of UGT1A were significantly associated with platelet count and total bilirubin level (p=0.01, p=0.01, respectively). Additionally, total bilirubin level was positively correlated with occurrence of the UGT1A9-118 $(T)_{9>10}$ rare variant. Common haplotypes encompassing six UGT1A polymorphisms were significantly associated with total bilirubin level (p=0.01). Taken together, we suggest that determination of the UGT1A1 and UGT1A9 genotypes is clinically useful for predicting the efficacy and serious toxicities of particular drugs requiring glucuronidation.
Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.
Kim Yong-Gyun;Basio Neil A.;Ibrahim Ahmed M.A.;Bae Sung-Woo
Korean journal of applied entomology
/
v.45
no.1
s.142
/
pp.15-24
/
2006
Inhibitor kB (IkB)-like gene has been found in the genome of Cotesia plutellae bracovirus (CpBV), which is the obligatory symbiont of an endoparsitoid wasp, C. plutellae. The open reading frame of CpBV-IkB was 417 bp and encoded 138 amino acids. Four ankyrin repeat domains were found in CpBV-IkB, which shared high homology with other known polydnavirus IkBs. Considering a presumptive cellular IkB based on Drosophila Cactus, CpBV-IkB exhibited a truncated structure with deletion of signal-receiving domains, which suggested its irreversible inhibitory role in NFkB signal transduction pathway of the parasitized host in response to the wasp parasitization. CpBV-IkB was expressed only in the parasitized diamondback moth, Plutella flostella. Its expression was estimated by quantitative RT-PCR during parasitization period, showing a constitutive expression pattern from the first day of parasitization. An indirect functional analysis of CpBV-IkB was conducted and suggested a hypothesis of host antivirus inhibition.
Journal of the Korean Society of Marine Environment & Safety
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v.18
no.5
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pp.379-387
/
2012
In order to see the oceanographic conditions, the observations of aquaculture farm of ascidian in Youngdeuk, the east coast of Korea were conducted through 6 times-23 February, 6 April, 8 June, 19 August, 6 October and 20 December-in 2010. Surveys were conducted in 20 stations bimonthly using SBE 19 CTD instrument. The mixed layer depth (MLD) was deep in winter and shallow in summer. The cold water below $5^{\circ}C$ in temperature was occupied below thermocline through all season. The temperature was high in the southeastern area. The salinity was increased from the coast to the open sea. The halocline was distinct at 20 m depth in August and at 40 m depth in October. The lowest value of salinity was appeared at the depth of 10 m in October. In addition the value of precipitation minus evaporation denoted negative in October. These low saline water seemed to inflow to the coast from the open sea. Therefore the low saline water moved to the east coast of Korea. The EKWC may play an important role to convey the low saline water. It may affect the aquaculture farm along the coast as the mass mortality of ascidian. It needs to clarify the role and pathway of EKWC to transfer the low saline water along the east coast of Korea.
It has widely been observed that the effect of elevating atmospheric $CO_2$ concentrations on rice productivity depends largely on soil N availabilities. However, the responses of ammonia volatilization from flooded paddy soil that is an important pathway of N loss and thus affecting fertilizer N availability to concomitant increases in atmospheric $CO_2$ and temperature has rarely been studied. In this paper, we first report the interactive effect of elevated $CO_2$ and temperature on ammonia volatilization from rice paddy soils applied with urea. Urea labeled with $^{15}N$ was used to quantitatively estimate the contribution of applied urea-N to total ammonia volatilization. This study was conducted using Temperature Gradient Chambers (TGCs) with two $CO_2$ levels [ambient $CO_2$ (AC), 383 ppmv and elevated $CO_2$ (EC), 645 ppmv] as whole-plot treatment (main treatment) and two temperature levels [ambient temperature (AT), $25.7^{\circ}C$ and elevated temperature (ET), $27.8^{\circ}C$] as split-plot treatments (sub-treatment) with triplicates. Elevated temperature increased ammonia volatilization probably due to a shift of chemical equilibrium toward $NH_3$ production via enhanced hydrolysis of urea to $NH_3$ of which rate is dependent on temperature. Meanwhile, elevated $CO_2$ decreased ammonia volatilization and that could be attributed to increased rhizosphere biomass that assimilates $NH_4^+$ otherwise being lost via volatilization. Such opposite effects of elevated temperature and $CO_2$ resulted in the accumulated amount of ammonia volatilization in the order of ACET>ACAT>ECET>ECAT. The pattern of ammonia volatilization from applied urea-$^{15}N$ as affected by treatments was very similar to that of total ammonia volatilization. Our results suggest that elevated $CO_2$ has the potential to decrease ammonia volatilization from paddy soils applied with urea, but the effect could partially be offset when air temperature rises concomitantly.
Background & Objective : Naeso-san(NSS) has been used for the treatment of functional dyspepsia, regarded as a gastric dysmotility disease. A main cause of gastric dysmotility is antral dilatation or antroduodenal uncoordination. Therefore, we investigated the effect of NSS on gastric motility and its mechanism of action, as well as the morphologic changes in antral dilatated rats. Methods : Antral dilatated rats were induced by wrapping a nonabsorbable rubber ring(D:6mm, W:4mm, T:1mm) around the 1st portion of the duodenum for 8 weeks. Then morphologic changes were investigated and compared with normal intact rats before and after 8 weeks. Gastric emptying was measured by administration of normal saline(NS) or NSS in normal intact and antral dilatated rats. In another series of experiments to evaluate the mechanism of NSS under delayed conditions, normal intact rats were treated with atropine sulfate(1mg/kg, s.c.), quinpirole HCl(0.3mg/kg, i.p.), $NAME(N^{G}-nitro-L-arginine$ methyl ester, 75mg/kg, s.c.) and cisplatin(10mg/kg, i.p.), respectively. The myoelectrical activity of the gastric smooth muscle was recorded in normal intact and antral dilatated rats. The contractile waves were measured for 30 minutes before and after administration of each solution(NS, NSS). Results : Body weight gain of antral dilatated rats was significantly lower than that of the controls. Futhermore, we found the thickness of the mucosal and muscular layers and surface area of the stomach increased significantly compared with controls. NSS 278㎎/㎏ improved gastric emptying more than normal saline or NSS 93mg/kg in normal intact(p=0.026) and antral dilatated rats(p=0.03). NSS enhanced gastric emptying significantly in the NAME treated group(p=0.002). NSS 278mg/kg increased the significant postprandial dominant power than that of NS in normal intact rats, whereas there was no statistical significance in antral dilatated rats. Conclusions : NSS stimulates gastric motility through the cholinergic pathway. We expect that pathologic model with antral dilatation can be used as an exprimental tool which is similar to dyspepsia and NSS would be effective especially in dysmotility-like functional dyspepsia with antral dilatation or impaired reservoir functions such as gastric adaptive relaxation.
Polysaccharides isolated from Korean black raspberry wine were examined for their chemical properties and immuno-modulating activities. The molecular mass of RB-1b-I, the main polysaccharide in black raspberry wine, was estimated as 180 kDa and it contained a significant proportion of mannose (76.8%) and 4 different minor component sugars such as galactose (15.8%), arabinose (3.8%), glucose (2.6%) and rhamnose (1.2%). This indicated that RB-1b-I was mainly present as a mannan, which had originated from the cell walls of fermenting yeasts. On the other hand, RB-1b-I induced high levels of macrophage activation as well as mitogenicity regarding murine splenocytes in vitro. The intravenous administration of RB-1b-I significantly augmented NK cytotoxicity against YAC-1 tumor cells. RB-1b-I also showed potent anti-complementary activity in a dose-dependent manner via both alternative and classical pathways. Results indicated that Korean black raspberry wine contains peculiar polysaccharides which provide beneficial immuno-stimulating activities for human health.
Yeast pheromone a-factor is a 13-amino acid peptide hormone that is synthesized as a part of a larger precursor, prepro-$\alpha$-factor, consisting of a signal peptide and a proregion of 64 amino acids. The carboxy-terminal half of the precursor contains four tandem copies of mature $\alpha$-factor. To investigate the molecular basis of intracellular sorting, proteolytic processing, and storage of the peptide hormone, yeast prepro-$\alpha$-factor precursors were heterologously expressed in rat pituitary $GH_3 cells. When cells harboring the precursor were metabolically labeled, a species of approximately 27 kD appeared inside the cells. Digestion with peptide: N-glycosidase F (PNG-F) shifted the molecular mass to a 19 kD, suggesting that the 27 kD protein was the glycosylated form as in yeast cells. The nascent polypeptide is efficiently targeted to the ER in the $GH_3 cells, where it undergoes cleavage of its signal peptide and core glycosylation to generate glycosylated pro-a-factor. To look at the post ER intracellular processing, the pulse-labelled cells were chased up to 2 hrs. The nascent propeptides disappeared from the cells at a half life of 30 min and only 10-25% of the newly synthesized, unprocessed precursors were stored intracellularly after the 2 h chase. However, about 20% of the pulse-labeled pro-$\alpha$-factor precursors were secreted into the medium in the pro-hormone form. With increasing chase time, the intracellular level of propeptide decreased, but the amount of secreted propeptide could not account for the disappearance of intracellular propeptide completely. This disappearance was insensitive to lysosomotropic agents, but was inhibited at $16^{circ}C or 20^{\circ}C$, suggesting that the turnover of the precursors was not occurring in the secretory pathway to trans Golgi network (TGN) or dependent on acidic compartments. From these results, it is concluded that a pan of these heterologous precursors may be processed at its paired dibasic sites by prohormone processing enzymes located in TGN/secretpry vesicles producing small peptides, and that the residual unprocessed precursors may be secreted into the medium rather than degraded intracellularly.
Jo, Mi Na;Jung, Ji En;Yoon, Hyun Joo;Chang, Kyung Hoon;Jee, Hee Sook;Kim, Kee-Tae;Paik, Hyun-Dong
Microbiology and Biotechnology Letters
/
v.42
no.4
/
pp.347-353
/
2014
${\beta}$-Glucosidase from Aspergillus usamii KCTC 6954 was used to convert ginsenoside Rb1 to compound K, which has a high bio-functional activity. The enzymatic activities during culturing for 15 days were determined using ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside. The growth rate of the strain and the enzymatic activity were maximized after 6 days (IU; $175.93{\mu}M\;ml^{-1}\;min^{-1}$). The activities were maximized at $60^{\circ}C$ in pH 6.0. During culturing, Rb1 was converted to Rd after 9 d and then finally converted to compound K at 15 d. In the enzymatic reaction, Rb1 was converted to the ginsenoside Rd within 1 h of reaction time and compound K could be detected after 8 h. As a result, this study demonstrates that $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K is the main metabolic pathway catalyzed by ${\beta}$-glucosidase and that ${\beta}$-glucosidase is a feasible option for the development of specific bioconversion processes to obtain minor ginsenosides such as Rd and compound K.
Exposure to high temperatures during the reproductive period of crops decreases their productivity. The Intergovernmental Panel on Climate Change's (IPCC) fifth Assessment Report predicts that the frequency of high temperatures will continue to increase in the future, resulting in significant impacts on the world's food supply. This study evaluate climate change-induced heat stress on four major agricultural crops (rice, maize, soybean, and wheat) at a global level, using the coupled atmosphere-ocean model of Hadley Centre Global Environmental Model version 2 (HadGEM2-AO) and FAO/IIASA Global Agro-Ecological Zone (GAEZ) model data. The maximum temperature rise ($1.8-3.5^{\circ}C$) during the thermal-sensitive period (TSP) from the baseline (1961-1990) to the future (2070-2090) is expected to be larger under a Representative Concentration Pathway (RCP) 8.5 climate scenario than under a RCP2.6 climate scenario, with substantial heat stress-related damage to productivity. In particular, heat stress is expected to cause severe damage to crop production regions located between 30 and $50^{\circ}N$ in the Northern Hemisphere. According to the RCP8.5 scenario, approximately 20% of the total cultivation area for all crops will experience unprecedented, extreme heat stress in the future. Adverse effects on the productivity of rice and soybean are expected to be particularly severe in North America. In Korea, grain demands are heavily dependent on imports, with the share of imports from the U.S. at a particularly high level today. Hence, it is necessary to conduct continuous prediction on food security level following the climate change, as well as to develop adaptation strategy and proper agricultural policy.
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