• Journal of Pharmaceutical Investigation
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• v.36 no.5
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• pp.331-338
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• 2006

A rapid, selective and sensitive reverse-phase HPLC methods for the determination of atenolol and chlorthalidone in human serum and whole blood were validated, and applied to the pharmacokinetic study of atenolol and chlorthalidone combination therapy. Atenolol and an internal standard, pindolol, were extracted from human serum by liquid-liquid extraction, and analyzed on a $\mu$-Bondapak C18 $10-{\mu}$ column in a mobile phase of methanol-0.01 M potassium dihydrogenphosphate(30:70, v/v, adjusted to pH 3.5) and fluorescence detection(emission: 300 nm, excitation: 224 nm). Chlorthalidone and an internal standard, probenecid, were extracted form human whole blood by liquid-liquid extraction, and analyzed on a Luna C18 $5-{\mu}$ column in a mobile phase of acetonitrile containing 77% 0.01 M sodium acetate and UV detection at 214 nm. These analysis were performed at three different laboratories using the same quality control(QC) samples. The chromatograms showed good resolution, sensitivity, and no interference by human serum and whole blood, respectively. The methods showed linear responses over a concentration range of 10-1,000 ng/mL for atenolol and 0.05-20 ${\mu}g/mL$ for chlorthalidone, with correlation coefficients of greater than 0.999 at all the three laboratories. Intra- and inter-day assay precision and accuracy fulfilled international requirements. Stability studies(freeze-thaw, short-, long-term, extracted sample and stock solution) showed that atenolol and chlorthalidone were stable. The lower limit of quantitation of atenolol and chlorthalidone were 10 ng/mL and 0.05 ${\mu}g/mL$, respectively, which was sensitive enough for pharmacokinetic studies. These methods were applied to the pharmacokinetic study of atenolol and chlorthalidone in human volunteers following a single oral administration of Hyundai $Tenoretic^{\circledR}$ tablet(atenolol 50 mg and chlorthalidone 12.5 mg) at three different laboratories.

• Applied Biological Chemistry
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• v.38 no.5
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• pp.382-386
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• 1995

The potato proteinase inhibitor II (PI-II) protein contains chymotrypsin and trypsin inhibitory site. Among several PI-II genes isolated from genomic library, amino acid sequence deduced from PI-IIT gene has 84% identity with that of the polypeptide chymotrypsin inhibitor (PCI). Therefore a gene fragment having homology with the PCI was cloned into a vector using polymerase chain reaction(PCR) from the potato proteinase inhibitor IIT gene. Two different primers were utilized for cloning; primer A contains NdeI restriction site and 30 nucleotides, which has AUG N-terminal methionine codon, primer B contains BclI restriction site and 28 nucleotides, which has TAG translation stop codon. After PCR, about 160 bp-long DNA fragment was cloned into pRT146, derivative of pUC118, and sequenced. The sequenced NdeI/BclI fragment was moved to pET3a, containing bacteriophage T7 promoter and terminator. The expressed proteins in E. coli BL2l(DE3) were determined on a polyacrylamide gel containing sodium dodecyl sulfate. The expected size of protein deduced from the sequenced gene fragment is about 6,500 dalton whose size was similar to the IPTG-induced protein (6,000 dalton) on a gel. However the expression level was much lower than expected.

• Applied Biological Chemistry
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• v.17 no.4
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• pp.257-274
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• 1974

The studies on the carotenoids in the viscera of abalone (Haliotis discus hannai) have been-carried out. The pigments were extracted with acetone-methanol mixture (4 : 1) from the viscera of abalones which were caught around the coastal water of Korea from March to August. The individual carotenoid in the extracts was separated and purified by the silica gel TLC, $Mg(OH)_2$ impregnated paper chromatography and $Mg_2(OH)_2CO_3$ TLC. The isolated eleven carotenoids were investigated and identified by epoxide test, partition test, reduction with sodium borohydride, alkaline hydrolysis, co-chromatography and comparative test with reference carotenoids and electronic and IR absorption spectrophotometry. ${\alpha}$-Carotene, ${\beta}$-carotene. lutein, zeaxanthin, siponaxanthin, siponein, fucoxanthin, loroxan-thin-like and fucoxanthinol-like have been identified among the eleven carotenoids isolated. It has been found that fucoxanthin, on alkaline treatment, was transformed to the product of which chromophore was the same one as fucochrome and semifucoxanthol. Among the identified nein carotenoids siphonaxanthin, siponein, fucoxanthin, loroxanthin-like and fucoxanthinol-like have not been reported previously to be contained in the shellfish.

• Applied Biological Chemistry
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• v.35 no.2
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• pp.76-81
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• 1992

The technicals of KC-7079, isoprothiolane, perfluidone and tricyclazole were granulated with a mixture of inorganic carrier and oil-soluble binder, that is, stearly alcohol or ethyl cellulose. The concentration of the released active ingredient from the granules was analyzed at several days intervals after immersion of these granules in water at $25^{\circ}C$. At the content of stearyl alcohol less than $80g\;kg^{-1}$, the granule kneaded with stearyl alcohol mixture and water disintegrated in water. But the granule kneaded with methanol disintegrated in water at the content of stearyl alcohol less than $30g\;kg^{-1}$. The less the KC-7079-stearyl alcohol granule disintegrated, the slower the release rate of KC-7079 was. No matter how was increased the stearyl alcohol content, the release rate of KC-7079 granule which did not disintegrate was not significantly changed. The sustained releasing effect of the granules was little in the other three pesticides of which the water solubility was higher than of KC-7079(21 ppm). The granule made of ethyl cellulose did not disintegrate even at $5g\;kg^{-1}$ of ethyl cellulose. With the increase of ethyl cellulose content and the decrease of active ingredient in the granules, the sustaining effect of the granules on releasing acitive ingredient was increased. The lower the water solubility of pesticide was, the release rate tended to be sustained except perfluidone.

• Applied Biological Chemistry
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• v.29 no.1
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• pp.83-89
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• 1986

To investigate effectors on the colloidal stability of whey and soybean proteins, characteristics of tofu-gel formation, effects of heat treatment and salt composition on the colloidal stability, and effects of heat treatment on storage stability were analyzed. When experimental tofus were made from the mixture of whey and soybean, the calcium in the whey precipitated the soy proteins, and disrupted the gel formation, which resulted in the curd of poor texture. In the heat treatment at $60{\sim}100^{\circ}C$, whey and the whey proteins dialyzed against distilled water were readily preciptated at over $70^{\circ}C$, but the mixture of whey and soy extract as well as soy extract were stable at the range of temperature. The proteins of soy extract, whey dialyzed against sodium phosphate buffer, and the mixture were stable at the same heat treatment, and this suggested that phosphates in the soy extract stabilize specialty the whey proteins. Soy proteins were easily destabilized by adding $CaCl_2(0.05{\sim}0.07M)$ at $80{\circ}C$ and $70{\sim}85%$ of the proteins in soy extract and the mixture were preciptated, while only $30{\sim}55%$ of the proteins in whey dialyzed against distilled water were destabilized at the same conditions. Storage stability at $4^{\circ}C$ of the mixture was increased when the mixture was treated at $63^{\circ}C$ and lower temperature.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.5
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• pp.427-434
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• 2015

Significant evidence supports the role of the vestibular system in the regulation of blood pressure during postural movements. In the present study, the role of the vestibulo-spino-adrenal (VSA) axis in the modulation of blood pressure via the vestibulosympathetic reflex was clarified by immunohistochemical and enzyme immunoassay methods in conscious rats with sinoaortic denervation. Expression of c-Fos protein in the intermediolateral cell column of the middle thoracic spinal regions and blood epinephrine levels were investigated, following microinjection of glutamate receptor agonists or antagonists into the medial vestibular nucleus (MVN) and/or sodium nitroprusside (SNP)-induced hypotension. Both microinjection of glutamate receptor agonists (NMDA and AMPA) into the MVN or rostral ventrolateral medullary nucleus (RVLM) and SNP-induced hypotension led to increased number of c-Fos positive neurons in the intermediolateral cell column of the middle thoracic spinal regions and increased blood epinephrine levels. Pretreatment with microinjection of glutamate receptor antagonists (MK-801 and CNQX) into the MVN or RVLM prevented the increased number of c-Fos positive neurons resulting from SNP-induced hypotension, and reversed the increased blood epinephrine levels. These results indicate that the VSA axis may be a key component of the pathway used by the vestibulosympathetic reflex to maintain blood pressure during postural movements.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.150-155
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• 1988

In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.893-902
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• 2011

Endophytic fungi are little known for their role in gibberellins (GAs) synthesis and abiotic stress resistance in crop plants. We isolated 10 endophytes from the roots of field-grown soybean and screened their culture filtrates (CF) on the GAs biosynthesis mutant rice line - Waito-C. CF bioassay showed that endophyte GMH-1B significantly promoted the growth of Waito-C compared with controls. GMH-1B was identified as Penicillium minioluteum LHL09 on the basis of ITS regions rDNA sequence homology and phylogenetic analyses. GC/MS-SIM analysis of CF of P. minioluteum revealed the presence of bioactive $GA_4$ and $GA_7$. In endophyte-soybean plant interaction, P. minioluteum association significantly promoted growth characteristics (shoot length, shoot fresh and dry biomasses, chlorophyll content, and leaf area) and nitrogen assimilation, with and without sodium chloride (NaCl)-induced salinity (70 and 140 mM) stress, as compared with control. Field-emission scanning electron microcopy showed active colonization of endophyte with host plants before and after stress treatments. In response to salinity stress, low endogenous abscisic acid and high salicylic acid accumulation in endophyte-associated plants elucidated the stress mitigation by P. minioluteum. The endophytic fungal symbiosis of P. minioluteum also increased the daidzein and genistein contents in the soybean as compared with control plants, under salt stress. Thus, P. minioluteum ameliorated the adverse effects of abiotic salinity stress and rescued soybean plant growth by influencing biosynthesis of the plant's hormones and flavonoids.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.393-401
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• 2014

A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.4
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• pp.679-688
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• 2000

Dispersants, which are used to break water-in-oil emulsions and to remediate oil-spills, are another water pollutants. In this study, magnetic separation technology was applied to remove dispersants from the sea. Magnetite and maghemite were used as magnetic sorbents and SDDBS, an anionic surfactant and Triton X-100, a nonionic surfactant, were employed as dispersants. Batch experiments were undertaken to study the sorption capacity and sorption equilibrium, and water-bath experiments were conducted to simulate the real situation and to describe the recovery of magnetic particles by the permanent magnet or electromagnet. Maghemite has rather constant removal efficiency for dispersants, regardless of surfactant species. On the other hand, removal efficiency by magnetite is higher for anionic surfactant than maghemite and is higher in distilled water than in seawater which contains more ions. The sorption of dispersants to magnetite is explained by electrostatic attraction and that of maghemite is described not only by electrostatic attraction, but also by structural characteristics that provide high sorption ability and surface condition. Water bath experimental results showed that recovery efficiency of magnetic particle after sorption for dispersants is nearly 100%. It is suggested that this magnetic separation technology is an effective way of dispersant removal because of short operating time, high sorption capacity, and high recovery efficiency of sorbents.