• Title/Summary/Keyword: Selective media

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Cell fusion and fusants characterization of Bacillus strains producing biopolymer (Biopolymer 생산성 Bacillus 속 균주의 세포융합과 융합주의 세포특성)

  • Yim, Moo-Hyun;Kim, Seong-Ho
    • Applied Biological Chemistry
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    • v.42 no.1
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    • pp.12-19
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    • 1999
  • To improve biopolymer productivity and properties of Bacillus strains, protoplast fusion was performed between biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. The results were as follows; Protoplasts mixed in fusion fluid containing 33% PEG 6000, 1% PVP and 10 mM $CaCl_2$ were reacted for 5 min at $37^{\circ}C$ and then centrifused protoplasts were directly overlaid on the selective media containing $100\;{\mu}g/ml$ antibiotics and incubated for 3 days. At this conditions, the frequency of protoplast fusion was generally in the range of $4.6{\times}10^{-5}\;to\;1.8{\times}10^{-7}$ in ratio. Segregation ratio was observed between 1 to 6% indicating genetic stability of all the fusants. Fusants growth were also observed on the media contained amino acid and antibiotics as required marked materials. DNA contents of the selected fusants were 1.6 to 2.7 times more than that of parent strains. With observation by TEM microscopy, spherical protoplasts were first released from the swollen parental cells and then contracted to fuse in the process of fusion. And fused cells were observed representative vesicle. Originally, the parental cells were observed as in the morphology of thick-walled and double membrane-surrounded rod shape with TEM microscopy.

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Isolation and Characterization of Marine Bacterial Strain SH-1 Producing Agar-Degrading Enzymes (한천 분해효소를 생산하는 해양 미생물 SH-1의 분리 및 특성 분석)

  • Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.324-330
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    • 2014
  • A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

Bioequivalenee of Samchundang Atenolol Tablet to Tenolmin Tablet (테놀민 정에 대한 삼천당아테놀올 정의 생물학적동등성)

  • Cho, Hea-Young;Kang, Hyun-Ah;Lee, Suk;Baek, Seung-Hee;Lee, Yong-Bok
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.339-344
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    • 2003
  • Atenolol is a water soluble, ${\beta}_1$ selective adrenoceptor antagonist used in the treatment of angina and hypertension. It is primarily eliminated renally with minimal hepatic metabolism. The purpose of the present study was to evaluate the bioequivalence of Samchundang Atenolol (Samchundang Pharmaceutical Co., Korea.) to Tenolmin(Hyundai Pharmaceutical Ind. Co., Korea). The atenolol release from the two atenolol tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty four normal male volunteers, 22.83$\pm$1.99 years in age and 65.82$\pm$7.15 kg in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 50 mg of atenolol was orally administered, blood was taken at predetermined time intervals and the concentrations of atenolol in serum were determined using HPLC method with fluorescence detector. The dissolution profiles of two atenolol tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_{t}$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_{t}$ and $C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AUC_{t}$, $C_{max}$ and $T_{max}$ between two tablets based on the Tenolmin were 3.74%, 4.38% and 17.77%, respectively. There were no sequence effects between two tablets in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) (e.g., log(0.98)∼log(1.l1) and log(0.95)∼log(1.l5) for $AUC_{t}$ and $C_{max}$ respectively), indicating that Samchundang Atenolol tablet is bioequivalent to Tenolmin tablet.

A Correlation Study between the Environmental, Personal Exposures and Biomarkers for Volatile Organic Compounds (대기 중 휘발성유기오염물질의 환경, 개인 및 인체 노출의 상관성 연구)

  • Jo, Seong-Joon;Shin, Dong-Chun;Chung, Yong;Breysse, Patrick N.
    • Environmental Analysis Health and Toxicology
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    • v.17 no.3
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    • pp.197-205
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    • 2002
  • Volatile organic compounds (VOCs) are an important public health problem throughout the world. Many important questions remain to be addressed in assessing exposure to these compounds. Because they are ubiquitous and highly volatile, special techniques must be applied in the analytical determination of VOCs. Personal exposure measurements are needed to evaluate the relationship between microenvironmental concentrations and actual exposures. It is also important to investigate exposure frequency, duration, and intensity, as well as personal exposure characteristics. In addition to air monitoring, biological monitoring may contribute significantly to risk assessment by allowing estimation of absorbed doses, rather than just the external exposure concentrations, which are evaluated by environmental and personal monitoring. This study was conducted to establish the analytic procedure of VOCs in air, blood, urine and exhaled breath and to evaluate the relationships among these environmental media. The subjects of this study were selected because they are occupationally exposed to high levels of VOCs. Environmental, personal, blood, urine and exhalation samples were collected. Purge & trap, thermal desorber, gas chromatography and mass selective detector were used to analyze the collected samples. Analytical procedures were validated with the“break through test”, 'quot;recovery test for storage and transportation”,“method detection limit test”and“inter-laboratory QA/QC study”. Assessment of halogenated compounds indicted that they were significantly correlated to each other (p value < 0.01). In a similar manner, aromatic compounds were also correlated, except in urine sample. Linear regression was used to evaluate the relationships between personal exposures and environmental concentrations. These relationships for aromatic and halogenated are as follows: Halogen $s_{personal}$ = 3.875+0.068Halogen $s_{environmet}$, ($R^2$= .930) Aromatic $s_{personal}$ = 34217.757-31.266Aromatic $s_{environmet}$, ($R^2$= .821) Multiple regression was used to evaluate the relationship between exposures and various exposure deter-minants including, gender, duration of employment, and smoking history. The results of the regression model-ins for halogens in blood and aromatics in urine are as follows: Halogen $s_{blood}$ = 8.181+0.246Halogen $s_{personal}$+3.975Gender ($R^2$= .925), Aromatic $s_{urine}$ = 249.565+0.135Aromatic $s_{personal}$ -5.651 D.S ($R^2$ = .735), In conclusion, we have established analytic procedures for VOC measurement in biological and environmental samples and have presented data demonstrating relationships between VOCs levels in biological media and environmental samples. Abbreviation GC/MS, Gas Chromatography/Mass Spectrometer; VOCs, Volatile Organic Compounds; OVM, Organic Vapor Monitor; TO, Toxic Organicsapor Monitor; TO, Toxic Organics.

Isolation of Hypervirulent Agrobacterium spp from Korea and Application for Transformation of Tobacco (한국산 고감염 Agrobacterium spp의 분리 및 연초의 형질전환에 이용)

  • 양덕춘;정재훈;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.207-217
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    • 1998
  • Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

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Bioequivalence Test of Fexofenadine Hydrochloride 120 mg Tablets (염산펙소페나딘 120밀리그람 정제의 생물학적동등성시험)

  • Cho, Hea-Young;Kang, Hyun-Ah;Kim, Se-Mi;Lee, Yong-Bok
    • YAKHAK HOEJI
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    • v.52 no.3
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    • pp.188-194
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    • 2008
  • Fexofenadine, ($\pm$)-4-1-hydroxy-4-{4-(hydroxydiphenylmethyl)-1-piperidinyl}-butyl-a,a-dimethyl benzeneacetic acid, is a selective histamine $H_1$ receptor antagonist, and is clinically effective in the treatment of seasonal allergic rhinitis and chronic idiopathic urticaria as a first-line therapeutic agent. The purpose of the present study was to evaluate the bioequivalence of two fexofenadine hydrochloride tablets, $Allegra^{(R)}$ (Handok Pharmaceuticals Co., Ltd.) and Alecort (Samchundang Pharmaceutical Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of fexofenadine from the two fexofenadine hydrochloride formulations in vitro was tested using KP VIII Apparatus II method with various dissolution media. Twenty six healthy male subjects, 25.62$\pm$3.35 years in age and 70.05$\pm$11.71 kg in body weight, were divided into two groups and a randomized 2$\times$2 cross-over study was employed. After a single tablet containing 120 mg as fexofenadine hydrochloride was orally administered, blood samples were taken at predetermined time intervals and the concentrations of fexofenadine in serum were determined using HPLC with fluorescence detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The harmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated, and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$, $C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Allegra^{(R)}$, were -1.37, 5.22 and 16.50% for $AUC_t$, $C_{max}$ and $T_{max}$, respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g., log 0.83$\sim$log 1.08 and log 0.81$\sim$log 1.03 for $AUC_t$ and $C_{max}$, respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Alecort tablet was bioequivalent to $Allegra^{(R)}$ tablet.

Effect of Ramulus et uncus uncariae on Glucose Oxidase-Induced Toxicity in Cultured Cerebral Neurons (조구등이 Glucose Oxidase로 손상된 대뇌신경세포에 미치는 효과)

  • Kim Hyeong Soo;Lee Yong Suk;Oh Suk Kyu;Lee Kang Chang;Lee Geon Mok;Lee Jeong;Lee Sang Bork;Kim Jong Ho;Yu Jun Ki;Kang Young Seong;Kim Sung Soo;Song Ho Jun;Park Seung Taeck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.1016-1019
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    • 2002
  • To examine the cytotoxic effect of glucose oxidase(GO) in cultured mouse cerebral neurons, cytotoxicity was measured by MTT assay after cultured nerve cells were incubated for 3 hours in the media containing 1 ~ 60mU/ml concentrations of GO. In addition, the neuroprotective effect of Ramulus et uncus uncariae(REUU) was determined by MTT assay in these cultrures. Cell viability was remarkably decreased in a dose- and time-dependent manner after cultured mouse cerebral neurons were exposed to 30mU/ml GO for 3 hours. In the neuroprotective effect of REUU on GO-induced toxicity, REUU blocked the GO-mediated neurotoxicity in these cultures. From above the results, it suggests that GO is toxic in cultured mouse cerebral neurons and selective herb extract such as REUU is effective in prevetion of the neurotoxicity induced by GO.

A Study on Resin Synthesis and Adsorption Characteristics for Separation and Recovery of U(VI) (우라늄(VI)의 분리회수를 위한 수지합성과 흡착특성에 관한 연구)

  • 강영식;노기환
    • Journal of environmental and Sanitary engineering
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    • v.14 no.1
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    • pp.31-41
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    • 1999
  • Several new ion exchange resins have been synthesized from chloromethyl styrene-1, 4-di-vinylbenzene with 1%, 2%, 5% and 10%-crosslinking and macrocyclic ligands of cryptand type by interpolymerization method. The adsorption characteristics and the pH, time, solvents and concentration dependence of the adsorption of metal ions by this resin were studied. The correlation between the separation characteristics of uranium and transition metal on the resins and the stability constants of complexes with macrocyclic ligands have been examined. The resins were very stable in both acidic and basic media and had good resistance to heat at $280^{\circ}C$. The $UO_2^{2+}$ aqueous solution was not adsorbed on the resins below pH 3.0, but the power of adsorption of $UO_2^{2+}$ increased rapidly above pH 4.0. The optimum equilibrium time for adsorption of metallic ions was twenty minutes and adsorptive power decreased in proportion to crosslinking size of the resins and order of dielectric constants of solvents used and the selective sequence for metal cations is in the order of $UO_2^{2+}$, $Cu^{2+}$ and $Ce^{3+}$ .

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Distribution of Pathogenic Vibrios in the Aquatic Environment Adjacent to Coastal Areas of South Korea and Analysis of the Environmental Factors Affecting Their Occurrence (2016년도 국내 해양환경내 병원성 비브리오균의 분포 및 해양환경인자간의 상관성 분석)

  • Jeong, Young-Il;Myung, Go-Eun;Choi, Eun-Jin;Soh, Sang-Moon;Park, Gi-Jun;Son, Tae-Jong
    • Journal of Environmental Health Sciences
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    • v.44 no.2
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    • pp.133-142
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    • 2018
  • Objectives: The pathogenic Vibrios genus denotes halophilic bacteria that are distributed in aquatic environments, including both sea and freshwater. Vibrio cholerae, Vibrio vulnificus, and Vibrio parahaemolyticus are the most important species since they can be potent human pathogens and leading causes of septicemia, wound infections, and seafood borne gastroenteritis. The recent emergence of a potential pandemic clone, V. cholera serotype O1 and the cholera outbreak in South Korea in 2016 indicates the importance of consistent surveillance of pathogenic Vibrio genus within coastal areas. Methods: The present study was undertaken to determine where and how vibrios live in the aquatic environment adjacent to coastal areas of South Korea. For this survey, a total of 838 samples were obtained at 35 different sites in South Korean coastal areas during the period from January 2016 to December 2016. Pathogenic vibrios was determined using the real-time PCR method, and its clones were isolated using three selective plating media. We also monitored changes in seawater and atmospheric temperature, salinity, turbidity, and hydrogen ion concentration at the collection points. Results: The total isolation rates of V. vulnificus, V. cholera (non-pathogenic, non-O1, non-O139 serogroups), and V. parahaemolyticus from seawater specimens in 2016 were 14.2, 13.48, and 67.06%, respectively. Conclusions: The isolation rates of pathogenic vibrios genus showed a positive correlation with temperature of seawater and atmosphere but were negatively correlated with salinity and turbidity.

A Study on the Adsorption of U(VI), NiI(II), Nd(III) Metal Ions Using Synthetic Resin (합성수지를 이용한 U(VI), NiI(II), Nd(III) 금속이온들의 흡착에 관한 연구)

  • 박성규;김준태;노기환
    • Journal of environmental and Sanitary engineering
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    • v.15 no.1
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    • pp.77-87
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    • 2000
  • Several new ion exchange resins have been synthesized from chloromethyl styrene-l,4-divinylbenzine with 1%, 2%, 10% and 20%-crosslink and macrocyclic ligands of cryptand type by interpolymerization method. The adsorption characteristics and the pH, time, solvents and concentration dependence of the adsorption of metal ions by this resin were studied. The correlation between the separation characteristics of uranium and transition metal on the resins and the stability constants of complexes with macrocyclic ligands have been examined. The resins were very stable in both acidic and basic media and have good resistance to heat. The $UO_2^{2+}$ was not adsorbed on the resins below pH 3.0, but the power of adsorption of $UO_2^{2+}$ increased rapidly above pH 4.0. The optimum equilibrium time for adsorption of metallic ions was two hours and adsorptive power decreased in proportion to crosslink size of the resins and order of dielectric constants of solvents used and the selective sequence for metal cations was in the order of $UO_2^{2+}$, $Ni{2+}$ and $Nd{3+}$.

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