• BMB Reports
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• v.29 no.5
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• pp.462-467
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• 1996

Acetolactate synthase (ALS, EC 4.1.3.18) is the first common enzyme in the biosynthesis of leucine, isoleucine, and valine. It is the target enzyme for several classes of herbicides, including the sulfonylureas, the imidazolinones, the mazolopyrimidines, the pyrimidyl-oxy-benzoates, the pyrimidyl-thio-benzens, and the 4,6-dimethoxypyrimidines. An amino-terminal fragment of the sulfonylurea-resistant ALS gene (SurB) from Nicotiana tabaccum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS1 was used to transform Escherichia coli strain BL21, and the tobacco ALS was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). Polyclonal antibodies against the fusion product (GST-ALS) were produced, and the sensitivity of GST-ALS with the rabbit anti-GST-ALS IgG was up to 50 ng. This antibody was used for Western blot analysis of the partially purified ALS from barley shoots. The results suggest that the polyclonal antibody produced in this study can be used to detect plant ALS.

• Clinical and Experimental Pediatrics
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• v.59 no.sup1
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• pp.116-120
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• 2016

Congenital hyperinsulinism (CHI) is a rare condition that can cause irreversible brain damage during the neonatal period owing to the associated hypoglycemia. Hypoglycemia in CHI occurs secondary to the dysregulation of insulin secretion. CHI has been established as a genetic disorder of islet-cell hyperplasia, associated with a mutation of the ABCC8 or KCNJ11 genes, which encode the sulfonylurea receptor 1 and the inward rectifying potassium channel (Kir6.2) subunit of the ATP-sensitive potassium channel, respectively. We report the case of a female newborn infant who presented with repetitive seizures and episodes of apnea after birth, because of hypoglycemia. Investigations revealed hypoglycemia with hyperinsulinemia, but no ketone bodies, and a low level of free fatty acids. High dose glucose infusion, enteral feeding, and medications could not maintain the patient's serum glucose level. Genetic testing revealed a new variation of ABCC8 mutation. Therefore, we report this case of CHI caused by a novel mutation of ABCC8 in a half-Korean newborn infant with diazoxide-unresponsive hyperinsulinemic hypoglycemia.

• Archives of Pharmacal Research
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• v.26 no.1
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• pp.9-14
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• 2003

For probing the importance of planarity of imidazolidinone motif of 4-phenyl-1-(benzenesulfonyl)imidazolidinones 1 for their cytotoxicity, 4-phenyl-2-(benzoyl)[1,2,5]thiadiazolidine-1,1-dioxide (2a), 4-phenyl-2-(p-toluoyl)[1,2,5]thiadiazolidine-1,1-dioxide (2b), 4-phenyl-2-(phenylcarbamoyl)[1,2,5]thiadiazolidine-1,1-dioxide (3a), and 4-phenyl-2-(p-tolylcarbamoyl)[1,2,5]thiadiazolidine-1,1-dioxide (3b) were prepared along with their regioisomers (5a, 5b, 9a, 9b) and their cytotoxicity were measured against human lung carcinoma (A549), human colon carcinoma (COLO205), human ovarian cancer (SK-OV-3), human leukemic cancer (K562), and murine colon adenocarcinoma (Colon26) cell lines in vitro. All compounds prepared do not show any activity against all five cancer cell lines unlike 1. Compounds 1 possess planarity of imidazolidinone, especially in sulfonylurea moiety ($-SO_2$NHCONH-). However compounds 2 and 3 have nonplanar 5-membered ring, [1,2,5]thiadiazolidine-1,1-dioxides. Such structural differentiation might result in the loss of activity. Therefore the inactivity of 2 and 3 could also be an indication for the necessity of planarity of imidazolidinone ring of 1 for their cytotoxic activity.

• Bulletin of the Korean Chemical Society
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• v.30 no.5
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• pp.1184-1186
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• 2009

• BMB Reports
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• v.28 no.6
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• pp.471-476
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• 1995

Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of valine, leucine, and isoleucine. ALS is the target enzyme for several classes of structually diverse herbicides. We have synthesized 4,6-dimethoxypyrimidine derivatives as ALS inhibitors, and their inhibitory activities on barley ALS were determined. $IC_{50}$ values for the derivatives are 0.2~200 ${\mu}m$. K11570, the most potent ALS inhibitor with $IC_{50}$ of 0.2 ${\mu}m$, showed mixed-type inhibition with respect to substrate pyruvate, and the progress curves for ALS inhibition by K11570 indicated that the amount of inhibition increased with time. Inhibition-competition experiments were carried out and indicated that three different classes of inhibitors, K11570, a sulfonylurea Ally, and leucine, bind to ALS in a mutually exclusive manner. Chemical modification of tryptophanyl and tyrosyl residues of ALS decreased the sensitivity of ALS to K11570, while cysteine modification did not affect the sensitivity. These results suggest that tryptophanyl and tyrosynyl residues are probably located at or near the inhibitor binding site.

• Journal of Pharmaceutical Investigation
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• v.27 no.3
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• pp.233-239
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• 1997

Glibenclamide is a second generation sulfonylurea that is orally active as a hypoglycemic drug. It exists as a crystalline powder which is sparingly soluble in water. It was investigated that the potential of glibenclamide to exhibit polymorphism. Three polymorphic modifications (form 1, form 2 and form 3) and three pseudopolymorphic modifications (form 4, form 5 and form 6) were obtained by crystallization from different organic solvents. The isolated crystal forms were characterized by differential scanning calorimetry(DSC), thermogravimetric analysis(TGA) and X-ray crystallography powder diffraction studies. Form 1 was the most stable and melt at $175.4^{\circ}C$. Form 2 was metastable and melt at $151.0^{\circ}C$. Form 3 was a new polymorphic modification because it was different from form 1 and form 2 in X-ray crystallography powder diffraction data. Form 4 was a 1 : 7(toluene : glibenclamide) toluene solvate; form 5 was a 1 : 5(toluene : glibenclamide) toluene solvate; form 6 was a 3 : 8(pentanol : glibenclamide) pentanol solvate. All forms were stable in 3-month storage under 0% or 100% humidity condition. The dissolution rate of form 4 was highest; those of form 2, form 3, form 1, form 5 and form 6 followed.

• Journal of Microbiology and Biotechnology
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• v.8 no.3
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• pp.295-299
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• 1998

Oligonucleotide primers were designed and successfully applied to amplify DNA fragments of P450 hydroxylase genes from actinomycetes which produce a large variety of medically important metabolites. Primers were designed based on several regions of strong similarities in amino acid sequence of P450 hydroxylases from a variety of actinomycetes, primarily in the regions of an oxygen binding site and a heme ligand pocket. These primers were used to amplify DNA fragments from seven different actinomycetes species producing a variety of different compounds. The deduced amino acid sequences of the isolated fragments revealed significant similarities to known P450 hydroxylase including the product of the suaC or subC genes from Streptomyces griseolus that is capable of metabolizing a number of sulfonylurea herbicides, and to the product of the $P450_{sca2}$ from S. carbophilus that produces a specific HMG-CoA reductase inhibitor. This method should help researchers in cloning the P450 hydroxylase genes involved in the biosynthesis of useful compounds.

• Korean Journal of Weed Science
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• v.31 no.2
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• pp.119-133
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• 2011

Sulfonylurea (SU)-resistant weeds include seven annual weeds such as Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and three perennial weeds of Scirpus planiculmis, Sagittaria pigmaea and Eleocharis acicularis as of 2010 since identification Monochoria korsakowii in the reclaimed rice field in 1998. The Echinochloa oryzoides resistant to acetyl CoA carboxylase (ACCase) and acetolactate synthase (ALS) inhibitors has been confirmed in wet-direct seeding rice field of the southern province, Korea in 2009. In the beginning of occurrence of SU-resistant weeds the M. vaginalis, S. juncoides and C. difformis were rapidly and individually spreaded in different fields, however, theses resistant weeds have been occurring simultaneously in the same filed as time goes by. The resistant biotype by weed species demonstrated about 10- to 1,000-fold resistance, base on $GR_{50}$ (50% growth reduction) values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. In history of paddy herbicides in Korea, the introduction of SU herbicides including besulfuron-metyl and pyrazosulfuron-ethyl that control many troublesome weeds at low use rates and provide excellent crop safety gave farmers and many workers for herbicide business refreshing jolt. The products and applied area of SU-included herbicides have been rapidly increased, and have accounted for about 69% and 96%, respectively, in Korea. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in direct-seeded rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which are effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. ALS genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006, the early period, in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly because the several resistant weeds have been occurring simultaneously in the same filed. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2 leaf stage.

• Korean Journal of Weed Science
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• v.12 no.3
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• pp.271-291
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• 1992

This study was conducted to survey the situation of direct rice seeding in Honam province in Korea to investigate problems and seek countermeasure of weed control in direct rice seeding. The total area of direct rice seeding in the south-western part of Korea (Chonbuk, Chonnam, and Chungnam) was 1650.8ha (732.1ha for direct seeding in dry field and 918.7ha for direct seeding in flooding field) in 1992. The followings are summary of the study. 1. In case of direct rice seeding in dry field, butachlor EC and G at 3 to 5 DAS was mostly selected by farmers to control weeds in dry field. Benthiocarb or chlornitrofen was also used in few cases. At 10 to 14 DAS just before rice emergence, tank misture of butachlor EC and paraquat was treated by some farmers. At 35 to 40 days, after flooding mixture of sulfonylurea derivatives was sequentially applied. Surviving weeds including barnyardgrass were finally controlled by mixture of bentazon+quinclorac WP foliage application. 2. In case of direct rice seeding in flooding field, weed control were mostly unsuccessful partially due to wrong selection of herbicide and missing the optimum application time. Three relatively successful weed control in the survey were summarized as follows. 1) Oxadiazon EC, butachlor or benthiocarb were treated just after puddling(5 to 7 days before seeding). then mixture of bentazone+quinclorac WP or sulfonylurea derivatives was sequently applied to control remaining weeds at 20 days after seeding. 2) Mixtures of bensulfuronmethyl+dimepiperate G, pyrazosulfuronethyl+molinate G, or bensulfuronmethyl+mefenacet+dymron G were applied at 11 days after puddling when barnyardgrass were at 2.0 leaf stage. Phytotoxicity was not found in case of mixture of bensulfuronmethyl+dimepiperate G but found in the other two cases but disappeared later. 3) Mixtures of bensulfuronmethyl+quinclorac G., pyrazosulfuronethyl+quinclorac G or betazone and quinclorac G were treated after 18 to 20 days after puddling when barnyardgrass was within 3.0 leaf stage. It showed good weed control in both annuals and perrenials without phytotoxicity. On the contrary, other sulfonylurea derivatives such as middle periodic herbicide showed poor weed control against barnyardgrass, so that sequential treatment of bentazone+quinclorac WP mixture was required. 3. Herbicidal characteristics and optimum application time of 45 rigistered herbicides in Korea were analyzed to discover new substitute for quinclorac mixture, that showed excellent weed control against barnyardgrass at its 3 leaf stage or older. The analysis revealed that 70% of herbicides were for preemergence and the others were post periodic herbicide. Most farmers favor to apply herbicide when rice seedlings completely rooted, at this time barnyardgrass are at 2.5-3.0 leaf stage. Therefore herbicide of which optimum application time had long is required. In this study. 6 middle periodic herbicides among sulfonylurea derivatives and 2 quinclorac mixture were selected and evaluated their weeding spectrums at different leaf stage of barnyardgrass in both soil application in flooding condition and foliage application in dry paddy field. The order of weeding spectrum in magnitude was as follows : bentazone+quinclorac WP> bentazone + quinclorac G>bensulfuronmethyl + quinclorac G>pyrazosulfuronethyl + quinclorac G> pyrazosulfuronethyl + Molinate G>bensulfuronmethyl + mefenacet + dymron G>bensulfuronmethyl + mefenacet G>bensulfuron methyl+benthiocarb G. The above results coincided with that of the survey. In conclusion, there is no proper substitute for quinclorac mixrure, which can control barnyardgrass at 3.0 leaf stage or even older. Therefore quinclorac should be supplied continuously to farmers in order to anchor direct rice seeding in Korea. Author suggested the followings to eastablish direct rice seeding technology effectively and quickly : 1) A tentatively named "The research committee for direct rice seeding" which was composed of farmers. researchers and goberment. should be eastablished to cooperate effectively. 2) Development of a pricise direct rice seeding machine for both dry and flooding paddy field. which is workable regardless of condition and varieties of seeds. 3) Study on protecting rice seed and seedling from sparrows. 4) Systematic studies of weed control techniques in direct rice seeding to standardize herbicide application. 5) Studies on farm-land reformation. techniques of precise land preparation. and direct rice seeding using an airplane.

• Korean Journal of Environmental Agriculture
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• v.22 no.1
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• pp.36-40
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• 2003

Cyclosulfamuron a herbicide of sulfonylurea type, is a relatively new compound which control broad leaves and perennial weeds in rice field. However, this herbicide has a minor disadvantage of decreasing rice plant growth, especially in early growth stage. Therefore, far introducing this cyclosulfamuron as a herbicide in rice field, it is important to minimize the suppression of early plant growth with maintaining weed control efficacy. This study was conducted to evaluate effects of cyclosulfamuron early plant growth and acetolactate synthase activity of rice (Oryza sativa cv Dongjinbyeo, Hwasungbyeo, Ilpumbyeo). Rice growth was inhibited by cyclosulfamuron in their early growth stage. The concentrations required far 50% inhibition of Dongjinbyeo, Hwasungbyeo and Ilpumbyeo growth were 6.3, 9.2 and 146.mg/kg, respectively. Inhibition effect of cyclosulfamuron on the root elongation was greater than the effect on the shoot growth. Concentrations required far 50% inhibition of acetolactate synthase activity from Dongjinbyeo, Hwasungbyeo, Ilpumbyeo were 42.7, 32.7 and $56.7\;{\mu}M$, respectively.