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Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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Adjuvant Effect of PAMAM Dendrimer on the Antigenicity of Keyhole Limpet Hemocyanin in Balb/c Mice (Balb/c 마우스에서 Keyhole limpet hemocyanine (KLH)의 항원성에 대한 PAMAM dendrimer 의 면역증강 효과)

  • Lee, Ga-Young;Kim, Min Jee;Kim, So Yeon;Lee, Kyung Bok;Oh, Dong Hyun;Cho, Young Ho;Yoo, Yung Choon
    • Journal of Life Science
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    • v.30 no.10
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    • pp.905-911
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    • 2020
  • The adjuvant effect of PAMAM dendrimer G4 (PAMAM) on the induction of humoral and cellular immune responses against keyhole limpet hemocyanin (KLH) was examined. Mice were immunized subcutaneously twice at two-week intervals with KLH, with or without PAMAM dendrimer (100 ㎍/mouse), and the mice immunized with KLH+PAMAM showed significantly higher antibody titers against KLH than those immunized with KLH alone. The assay for determining the isotypes of the antibodies showed that PAMAM augmented the KLH-specific antibody titers of IgG1, IgG2a, IgG2b, IgG3, and IgM. In addition, mice immunized twice with KLH+PAMAM followed by a subcutaneous injection of KLH (20 ㎍/site) 7 weeks after the primary immunization exhibited a higher delayed-type hypersensitivity (DTH) reaction than those treated with KLH alone. In an in vitro analysis of T lymphocyte proliferation in response to KLH in week 8, the splenocytes of mice treated with KLH+PAMAM showed significantly higher proliferating activity than those treated with KLH alone, and the culture supernatants of cell cultures from mice immunized with added PAMAM dendrimer showed higher levels of KLH-specific cytokine (IL-4 and IFN-r) production. These results suggest that PAMAM dendrimer G4 possesses a potent immune-adjuvant activity for enhancing both humoral and cell-mediated immunity specific to foreign antigens.

Photoalteration in Biodegradability and Chemical Compositions of Algae- derived Dissolved Organic Matter (자외선에 의한 조류기원 용존유기물의 생분해도 및 화학조성변환.)

  • Imai, Akio;Matsushige, Kazuo;Nagai, Takashi;Kim, Yong-Hwan;Kim, Bom-Chul;Choi, Kwang-Soon
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.235-241
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    • 2003
  • The effect of ultraviolet (UV) radiation on the characteristics of algae-derived dissolved organic matter (DOM) was examined by comparing the biodegradability and DOM fraction distribution of algal DOM before and after UV exposure. Algal DOM from two axenic cultures of Microcystis aeruginosa and Oscillatoria agardhii were irradiated for 24 h at a UV intensity of 42 W/$m^2$. A complete degradation of algal DOM during the UV exposure did not occur, remaining at constant concentrations of dissolved organic carbon(DOC). After UV exposure, however, microbial degradations were reduced by 17% in M. aeruginosa and 53% in O. agardhii, respectively, and decomposition rates also were two times lower in UV exposed algal DOM. In addition, the chemical compositions of algal DOM altered substantially after UV radiation exposure. The proportions of hydrophilic bases (HiB; protein-like DOM) decreased considerably in both algal DOM sources after UV exposure (16.8% and 20.0% of DOM, respectively), whereas those of hydrophilic acids (HiA; carboxylic acids-like DOM) increased as much as the decrease of the HiB fraction. Capillary ion electrophoresis (CE) analysis showed that several carboxylic acids increased significantly after UV exposure, further confirming an increase in HiA fractions. The results of this study clearly indicate that algal DOM can be changed in its chemical composition as well as biodegradability without complete degradation by UV radiation.

Production of Amylase by a Thermophi1ic Fungus, Mucor Sp. (고온성(高溫性) 사상균(絲狀菌) Mucor Sp.에 의(依)한 Amylase의 생산(生産))

  • Lee, Sang Ho;Park, Yoon Joong
    • Korean Journal of Agricultural Science
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    • v.15 no.2
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    • pp.153-163
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    • 1988
  • This experiment was carried out to obtain the thermophilic fungus producing amylase and to investigate properties of the amylase. The selected strain, L-11 was obtained from soil in the vicinity of a hot spring and identified as Mocor sp.. And then the conditions for enzyme production in wheat bran cultures and properties of the crude enzyme were investigated. Furthermore, the enzyme was purified and the characteristics of purified enzyme were studied. The results obtained were as follows: 1. On the wheat bran medium added 80-100% water, amylase was effectively produced by the selected strain, L-11 for 48 hrs incubation at $50^{\circ}C$. 2. When the crude enzyme solution of the strain L-11 was passed through DEAE-Sephadex A-50 column chromatography, two peaks having amylase activity were obtained, and one peak was that of the main enzyme (enzyme of B peak). 3. The purified enzyme (enzyme of B peak) was recognized as single protein band on polyacrylamide disc gel electrophoresis. 4. In the hydrolysis reaction of soluble starch by the enzyme of main amylase, oligosaccharides produced at early stage were maltose and maltotriose mainly and procedure of the reaction maltose amount of maltose and glucose was increased. 5. The strain L-11 was recognized as a special strain producing ${\alpha}-amylase$ mainly and scarcely glucoamylase. 6. The optimum pH, optimum temperature, pH stability, and temperature stability of ${\alpha}-amylase$ were pH 4.0, $60-65^{\circ}C$, pH 4.0-9.0, and below$70^{\circ}C$.

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The Hepatotprotective and Antioxidative Effects of Onion (Allium cepa) Extracts in Rat Hepatocyte Primary Culture (양파(Allium cepa) 추출물의 간보호 및 항산화 효과)

  • Lim Sang-Cheol;Rhim Tae-Jin
    • Korean Journal of Plant Resources
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    • v.18 no.3
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    • pp.470-478
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    • 2005
  • The objective of present study was to investigate the hepatoprotective and antioxidative effects of onion extracts. Primary cultures of rat hepatocytes were incubated with 1.5 mM tort-butyl hydroperoxide(t-BHP), potent oxidizing agent to liver, for 1 hr in the presence or absence of various concentrations (0, 0.01, 0.05, 0.1 or 0.3 mg/ml) of onion extract. Incubation with t-BHP increased glutamic oxaloacetic transaminase(GOT) and lactate dehydrogenase(LDH) acitivities and thiobarbituric acid reactive substances(TBARS) concentration but decreased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) reduction. Onion extracts at the concentration of 0.05 mg/ml decreased t-BHP-induced GOT and LDH activities. Onion extract at the concentration of 0.1 mg/ml increased t-BHP-induced MTT reduction. Onion extract at the concentration of 0.01 mg/ml decreased t-BHP-induced TBARS concentration. Taken together, onion extracts prevented t-BHP-induced hepatocyte injury and lipid peroxidation. Catalase, glutathione peroxidase(GSH-Px) and glutathione reductase(GSH-Rd) activities of hepatocytes were significantly decreased by t-BHP. Onion extracts at the concentration of 0.1 mg/ml prevented t-BHP-induced decrease in catalase, GSH-Px and GSH-Rd activities. Onion extracts prevented hydroxyl radical-induced single-strand breakage in dose-dependent manner when plasmid DNA was incubated with various concentrations of onion extracts in the presence of Fenton reagents producing hydroxyl radical. These results demonstrate that onion extracts suppressed t-BHP-induced cytoctoxicity, decreased viability and lipid peroxidation and increased GSH-Px, GSH-Rd and catalase activities. Thus hepatoprotective and antioxidant effects of onion extract seem to be due to, at least in part, the increase in antioxidant enzyme activities as well as prevention from hydroxyl radical-induced oxidation, followed by inhibition of lipid peroxidation.

The Manufactures and Characteristics of Chestnut Yoghurt (The Volatile Flavour Compounds and the Sensory Properties of Chestnut Yoghurt) (밤 Yoghurt의 제조와 특성 (밤 Yoghurt의 휘발성 향미성분 및 관능적 특성))

  • Shin, Hyeon-Su;Kim, Sang-Beom;Kim, Gi-Yeong;Ryu, Jin-Su;Lim, Jong-U
    • Journal of Dairy Science and Biotechnology
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    • v.19 no.1
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    • pp.4-12
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    • 2001
  • The effects of the addition of raw and dried chestnut with levels of 1%($T_1$),2%($T_2$),3%($T_3$) and 4%($T_4$) in skim milk substrate on the volatile flavour compounds and sensory properties of yoghurt fermented with the mixed cultures of YC-380 and ABT-4 during fermentation and storage period were investigated. In all treatments, the contents of acetaldehyde and acetone were detected at 2 hrs. and 1 hr. of fermentation, respectively and decreased with the storage period. The contents of ethanol and diacetyl were detected in all treatments at 3 hrs. and 4 hrs. of fermentation, respectively and increased significantly(p<0.05) with the storage period. The contents of volatile flavour compounds of treatments were increased gradually with decreasing the level of addition of raw and dried chestnut, and increased in order of fermented with YC-380 and ABT-4. The contents of volatile flavour compounds of raw chestnut yoghurt were slightly high compared to those of dried chestnut yoghurt. The taste, flavour and texture of $T_1$ were slightly higher than those of all treatments immediately after fermentation and during the storage period. The scores of sensory evaluation of treatments except $T_1$ were lowered significantly(p<0.05) with increasing the level of addition of raw and dried chestnut. The quality of both of flavour and texture, and taste were superior to chestnut yoghurt fermented with YC-380 and ABT-4, respectively. Generally the scores of sensory evaluation of dried chestnut yoghurt were slightly high compared to those of raw chestnut yoghurt. From the results mentioned above, the addition of raw and dried chestnut at 1%(w/v) level in skim milk substrate were suitable for volatile flavour compounds and sensory property of raw and dried chestnut yoghurt.

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Screening and Identification of a Cesium-tolerant Strain of Bacteria for Cesium Biosorption (환경유래의 세슘 저항성 균주 선별 및 세슘 흡착제거 연구)

  • Kim, Gi Yong;Jang, Sung-Chan;Song, Young Ho;Lee, Chang-Soo;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.4
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    • pp.304-313
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    • 2016
  • One of the issues currently facing nuclear power plants is how to store spent nuclear waste materials which are contaminated with radionuclides such as $^{134}Cs$, $^{135}Cs$, and $^{137}Cs$. Bioremediation processes may offer a potent method of cleaning up radioactive cesium. However, there have only been limited reports on $Cs^+$ tolerant bacteria. In this study, we report the isolation and identification of $Cs^+$ tolerant bacteria in environmental soil and sediment. The resistant $Cs^+$ isolates were screened from enrichment cultures in R2A medium supplemented with 100 mM CsCl for 72 h, followed by microbial community analysis based on sequencing analysis from 16S rRNA gene clone libraries(NCBI's BlastN). The dominant Bacillus anthracis Roh-1 and B. cereus Roh-2 were successfully isolated from the cesium enrichment culture. Importantly, B. cereus Roh-2 is resistant to 30% more $Cs^+$ than is B. anthracis Roh-1 when treated with 50 mM CsCl. Growth experiments clearly demonstrated that the isolate had a higher tolerance to $Cs^+$. In addition, we investigated the adsorption of $0.2mg\;L^{-1}$ $Cs^+$ using B. anthracis Roh-1. The maximum $Cs^+$ biosorption capacity of B. anthracis Roh-1 was $2.01mg\;g^{-1}$ at pH 10. Thus, we show that $Cs^+$ tolerant bacterial isolates could be used for bioremediation of contaminated environments.

Causative Organisms of Community Acquired Urinary Tract Infection and Their Antibiotic Susceptibility at a Secondary hospital in Korea (2차 병원에서의 지역 사회 획득 요로 감염의 원인균 및 항생제 감수성 분석)

  • Jo, Yun Ju;Lee, Eun Jeong;Choi, Kyong Min;Eun, Young Min;Yoo, Hwang Jae;Kim, Cheol Hong;Lee, Hyun Hee;Kim, Pyung Kil
    • Pediatric Infection and Vaccine
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    • v.17 no.1
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    • pp.30-35
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    • 2010
  • Purpose : We investigated the causative organism and its antibiotic susceptibility of community acquired urinary tract infection (UTI) in children at a secondary hospital to test the adequacy of the current guidelines. Methods : Children diagnosed with UTI at the Department of Pediatrics, Kwandong University MyMyongji Hospital by pyuria and bacterial growth of greater than $1.0{\times}10^5CFU/mL$ on clean catch midstream urine from January 2005 to December 2008 were studied retrospectively. The epidemiologic data, causative organism, and the antibiotic susceptibility were analyzed. Results : Sixty two children were diagnosed with sixty four cases of UTI's. Two bacteria were isolated in one case and thus data on 65 urine cultures were analyzed. The male:female ratio was 1.6:1 and 78.1% were less than 12 months of age. Escherichia coli was the predominant cause consisting of 53 cases (82.8%) of the cases. K. pneumoniae (5), Enterobacter (4), Enterococcus (1), $\beta$-streptococcus (1), Diphtheroides (1) were isolated. The antibiotic resistance of E. coli were as follows; ampicillin 69.8%, cefotaxime 1.9%, gentamicin 15.1%, amikacin 0.0%, levofloxacin 1.9%, and trimethoprim/sulfamethoxazole 26.4 %. Only one case of the E. coli was extended spectrum $\beta$-lactamase (ESBL) positive. Conclusion : Compared to prior reports from other tertiary hospitals in Korea, E. coli was the predominant cause in childhood UTI and the rate of ESBL positivity was low. The antibiotic resistance was also different compared to prior reports. We conclude that a difference in the cause and antibiotic resistance of childhood UTI exists between centers and this should be taken into consideration when prescribing antibiotics for childhood UTIs.

Colonization Rate and Control of Vancomycin-Resistant Enterococci in the Neonatal Intensive Care Unit (일개 신생아중환자실 반코마이신 저항 장구균(VRE)의 유행 양상과 조절)

  • Seo, Jung Ho;Nam, Ga Yeon;Park, Kyung Hee;Byun, Shin Yun;Park, Su Eun
    • Pediatric Infection and Vaccine
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    • v.17 no.1
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    • pp.1-8
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    • 2010
  • Purpose : Recently, vancomycin-resistant enterococci (VRE) have become one of the major nosocomial pathogens in Korea. However, there have been few studies on the epidemiology of VRE colonization among neonates. In this study, we investigated the prevalence of VRE colonization, risk factors for VRE, and how to control the spread of VRE infection in the Neonatal Intensive Care Unit (NICU) of Pusan National University Hospital (PNUH). Methods : We retrospectively reviewed medical records of 192 neonates who were admitted to the NICU of PNUH from March 2006 to March 2007. Surveillance cultures from rectal swabs for detecting VRE were obtained weekly during the study period. We analyzed the prevalence of VRE and various risk factors. Results : The rate of VRE colonization among NICU patients was 25% (48/192). Thirty five of these VRE colonized patients were transferred to the NICU from other local hospitals. Compared with the non-VRE group, the risk factors associated with VRE colonization were lower birth weight, congenital heart disease, applied mechanical ventilation, use of a central venous catheter, chest tubing, a history of surgery, and use of antibiotics. Conclusion : VRE colonization among patients admitted to the NICU is rapidly increasing. Monitoring and managing premature neonates from the beginning of the birth process, avoiding many invasive procedures, avoiding antibiotics such as vancomycin and third generation cephalosporin are important for preventing the emergence and spread of VRE colonization in the NICU.

Orbital and Periorbital Cellulitis in Children: A Recent 7-Year Clinical Review (소아에서 안와와 안와주위 봉와직염에 관한 최근 7년간의 임상적 고찰)

  • Choi, Jeong Ho;Hon, Sun Yeong;Park, Sung Sin;Cha, Sung Ho;Kim, Eun Jin;Lee, Jin;Chang, Jin Keun
    • Pediatric Infection and Vaccine
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    • v.15 no.1
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    • pp.36-44
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    • 2008
  • Purpose : Orbital cellulitis is rare, but it could be from the serious complication of sinusitis in children. It is often difficult to distinguish periorbital cellulitis from orbital cellulitis. The purpose of this study is to describe the clinical features of orbital and periorbital cellulitis in a pediatric population and to assess the predisposing factors and their complications. Methods : Forty-one patients aged 18 years and younger who were admitted between January 2000 and December 2006 to Hanil General Hospital and Kyunghee University Hospital with orbital or periorbital cellulitis. The retrospective analyses included clinical characteristics of orbital and periorbital cellulitis, dermographics, past history, predisposing factors, clinical presentations, treatments, and complications. Results : Among 41 patients, 34 patients had periorbital cellulitis, 7 patients had orbital cellulitis. While paranasal sinus disease was the most common predisposing cause in orbital cases, skin lesion, insect bite, dacrocystitis and conjunctivitis were the common causes in periorbital cases. In comparison with periorbital cases, orbital cases had higher level of white blood cell count, erythrocyte sedimentation rate, and C-reactive protein. Blood cultures were taken in 23 patients, but only one had Staphylococcus aureus from blood. Only one case had surgical incision and drainage and recovered without complications. Conclusion : From the results of our data, when patient shows erythematous swelling of periorbital area with opthalmoplegia, chemosis and proptosis, orbital CT scan is required to make diagnosis of orbital cellulitis. Most cases of orbital cellulitis can be treated successfully without surgical intervention. If there is no clinical improvement, repeated CT scan and/or surgical intervention should be considered.

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