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Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

DIFFERENCE IN BOND STRENGTH ACCORDING TO FILLING TECHNIQUES AND CAVITY WALLS IN BOX-TYPE OCCLUSAL COMPOSITE RESIN RESTORATION (박스 형태의 복합레진 수복시 충전법 및 와동벽에 따른 결합력 차이에 관한 연구)

  • Ko, Eun-Joo;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.34 no.4
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    • pp.350-355
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    • 2009
  • Bond strength depends on characteristics of bonding surface and restorative technique. The majority of studies dealing with dentin bond strength were carried out on flat bonding surface, therefore, difference of bond strength between axial wall and pulpal wall is not clear yet. This study evaluated bonding difference between cavity walls in class I composite resin restoration with different filling techniques. Twenty extracted caries-free human third molars were used. Cavities were prepared in 6 ${\times}$4 ${\times}$3 mm box-type and divided into four groups according to filling technique and bonding surface: Group I; bulk filling - pulpal wall, Group II; bulk filling - axial wall, Group III; incremental filling - pulpal wall, Group IV; incremental filling - axial wall. Cavities were filled with Filtek $Z250^{(R)}$(3M/ESPE., USA) and Clearfill SE $bond^{(R)}$(Kuraray, Japan). After 24 hour-storage in $37^{\circ}C$water, the resin bonded teeth were sectioned bucco-lingualy at the center of cavity. Specimens were vertically sectioned into 1.0 ${\times}$1.0 mm thick serial sticks perpendicular to the bond surface using a low-speed diamond saw (Accutom 50, Struers, Copenhagen, Denmark) under water cooling. The trimmed specimens were then attached to the testing device and in turn, was placed in a universal testing machine (EZ test, Shimadzu Co., Kyoto, Japan) for micro-tensile testing at a cross-head speed of 1 mm/min. The results obtained were statistically analyzed using 2-way ANOVA and t-test at a significance level of 95%. The results were as follows: 1. There was no significant difference between bulk filling and incremental filling. 2. There was no significant difference between pulpal wall and axial wall, either. Within the limit of this study, it was concluded that microtensile bond strength was not affected by the filling technique and the site of cavity walls.

Effect of Ortet Age and Types of Cuttings on Rooting, Cyclophysis and Topophysis of Rooted Cuttings in Taxus cuspidata S. et Z (주목삽수(揷穗)의 모수령(母樹齡) 및 아조형태(芽條形態)가 발근(發根)과 묘형(苗型)에 미치는 영향(影響))

  • Goo, Gwan Hyo;Lee, Kang Young;Youn, Ki Sik;Kwon, Yeong Han
    • Journal of Korean Society of Forest Science
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    • v.79 no.4
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    • pp.359-366
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    • 1990
  • This study was carried out to investigate rooting rate, plagiotrophic growth and root-promoting effect of IBA in Taxus cuspidata S, et Z. cuttings. The results obtained were summarized as follows ; 1. As the ortet ages of cuttings were increased in 5, 10, to 20 years, the rooting rates were decreased in order of 85.7%, 81.7% and 62.4%, and the number of primary roots were also decreased in 5.3, 3.7 and 2.9, respectively. 2. Rooting rate by cutting types of lateral shoot were higher than those of main shoot type, significant at the 1% level. 3. IBA(l00ppm) treatment promoted the rooting rates and the number of primary roots. 4. As the ortet ages were increased in 5, 10 to 20 years, angles of central shoot of rooted cuttings which mesured cyclophysis were decreased in order of $75.9^{\circ}$, $68.5^{\circ}$ and $59.6^{\circ}$, respectively, significant at the 1% level. 5. Average angles showed $77.0^{\circ}$ for main shoot cutting, $65.9^{\circ}$ for upper lateral shoot and $61.7^{\circ}$ for lower lateral, which was significant at 1% level, and lateral shoot cutting showed more topophysis than main shoot cutting. 6. Shoot length of rooted cuttings from main shoot cutting was longer than that from lateral shoot significant at the 1% level. 7. Relationship between some characteristics showed highly positive correlation.

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Effect of layer combinations with nanocomposite and low-shrinkage composite resins on their color and mechanical properties (나노복합레진과 저수축 복합레진의 복합 층으로 이룬 시편이 색과 물리적 성질에 미치는 영향)

  • Park, Wan-Ky;Choi, An-na;Son, Sung-Ae;Kwon, Yong Hoon;Kang, Eun-Sook;Park, Jeong-Kil
    • Korean Journal of Dental Materials
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    • v.44 no.2
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    • pp.129-139
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    • 2017
  • This study investigated the colors and mechanical properties of layered dental composites. Four nanocomposite resins (Aelite LS, Grandio, Tetric EvoCeram, Filtek Z350XT) and a silorane-based composite resin (P90) were used for overlying and underlying materials, respectively, with different thickness combinations. Colors, translucency parameter (TP), flexural and compressive properties were evaluated. All tested specimens had different color coordinates, although all were of A3 shade. Color coordinates and TP values of layered specimens better matched those of the corresponding overlying product as the thickness of the overlying product was increased. High TP values were related with high $b^*$ value differences between specimens (p<0.05). Both flexural strength and modulus, compressive strength and modulus of layered specimens with different thickness combinations were mostly lower than those of the corresponding overlying products, respectively, in their non-layered state.

Evaluation of Musculoskeletal Disease Risk Assessment and User Satisfaction in Accordance With Using Front-Entry Sitting Toilet for Wheelchair Users (휠체어 사용자의 전면진입착석 화장실 사용에 따른 근골격계질환 위험성 및 만족도 평가)

  • Park, Je-Mo;Hwang, Do-Yeon;Kim, Hee-Dong;Chung, Hyun-Ae;Jung, Hwa-Shik
    • Korean Journal of Occupational Therapy
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    • v.26 no.4
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    • pp.73-84
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    • 2018
  • Objective : The purpose of this study was to promote R&D of toilet design for the disabled by evaluating the Front-entry sitting toilet applied it to actual wheelchair users to examine the possibility of their application as a toilet that is low physical demanding and high satisfaction. Methods : Forty wheelchair users were asked to demonstrate using Front-entry sitting toilet after collecting the general characteristics and their current wheelchair usage information. Musculoskeletal disease risk assessment and user satisfaction were evaluated throughout Rapid Entire Body Assessment(REBA), Quebec Evaluation of Satisfaction with assistive Technology 2.0(QUEST) and subjective opinion questionnaire. Results : Statistical result showed that REBA scores($3.18{\pm}.38$) of Front-Entry Sitting Toilet were significantly lower(z=-5.930, p<.05) than the conventional toilet($6.53{\pm}1.15$). Overall, user satisfaction ratings were high with detailed scores were high in order of durability($4.48{\pm}.62$), safety($4.38{\pm}.63$), necessity($4.33{\pm}.73$) and universal ($4.6{\pm}.61$). Subjective opinion survey showed most were 'it was convenient to not have to rotate(30.8%).' Conclusion : This study found that Front-entry sitting toilet was highly likely to reduce the risk of musculoskeletal disorders and users' satisfactions were high. Through this study, we hope to expand the scope of occupational therapists and contribute to the physical, mental and social health in the daily activities of the people by applying various approaches.

Change in the Fish Fauna and Fish Community Characteristics in the Upper Reaches of the Seomgang (River), Korea (섬강 상류의 어류상 변화 및 어류군집 특성)

  • Hyeong-Su Kim;Mee-Sook Han;Myeong-Hun Ko
    • Korean Journal of Environment and Ecology
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    • v.38 no.3
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    • pp.246-262
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    • 2024
  • The survey conducted from 2018 to 2020 aimed to investigate the changes in fish fauna and community characteristics in the upper reaches of the Seomgang River, Korea. During the survey period, 35 sites were selected, resulting in the collection of 7,817 fish belonging to 12 families and 40 species. The dominant species was Zacco koreanus, with a relative abundance of 34.5%, followed by Z. platypus at 28.7%. Other significant species included Rhynchocypris oxycephalus (10.2%), Pungtungia herzi (5.3%), and Squalidus gracilis majimae (4.3%). Notably, four protected species - Acheilognathus signifer, Gobiobotia brevibarba, and Cottus koreanus, designated as class II endangered wildlife by the Ministry of Environment- were identified. These species predominantly inhabit the middle and lower reaches, except for Gobiobotia brevibarba, which is found in the upper reaches. Nineteen species, accounting for a 47.5% endemism rate, were endemic to Korea. The study also noted the presence of one climate-sensitive species, Cottus koreanus, and two exotic species, Carassius cuvieri and Micropterus salmoides. Community analysis indicated a trend of decreasing dominance and increasing diversity and richness from upstream to downstream, with a distinct division into uppermost reaches, upper reaches, middle and lower reaches, and lakes. The construction of the Hwaseong Dam has had a significant direct and indirect impact on the fish community. The habitat and abundance of endangered species such as R. pseudosericeus, A. signifer, and G. brevibarba decreased dramatically immediately after the dam's construction, transforming the submerged area from lotic to lentic environments. Approximately 20 years later, the habitats have stabilized, leading to an increase in the fish population and a recovery of the previously diminished endangered species. The river health (FAI) was also evaluated, with 27 sites rated as very good (A), seven as good (B), and one as fair (C). However, endangered species such as A. signifer continue to face threats from dam and river construction, while C. Koreanus has experienced a severe population decline due to river works. Additionally, the presence of the ecosystem-disrupting species M. salmoides in Hwaseong Lake raises concerns. To ensure a stable habitat for fish in the upper reaches of the Seomgang River, it is crucial to avoid indiscriminate river construction, urgently implement restoration policies for endangered species such as A. signifer, and develop management strategies to control the spread of invasive species such as bass.

Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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Synthesis and Spectroscopic Investigations of Some New rganotelluronium alts Based on Dicyclohexyl Telluride (디시클로헥실 텔루르염에 기반한 유기텔루로늄염의 성과 분광학적 분석)

  • Al-Rubaie, Ali Z.;Al-Mudhaffar, Dhafir M. H.;Al-Mowali, Ali H.;Asker, Kahtan A.
    • Journal of the Korean Chemical Society
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    • v.53 no.5
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    • pp.530-535
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    • 2009
  • Dicyclohexyl telluride was obtained in a high yield by the reaction of cyclohexyl bromide with NaTeH(prepared in situ) in an aqueous ethanolic solution. A series of new organotelluronium salts of the general formula ${(cyclo-C_6H_11)}_2Te(R)X$ (where R = $CH_3$, X = I (1); R = $C_2H_5$, X = Br(2); R = $C_2H_5$, X = I (3); R = C_3H_5$, X = Br (4)) were prepared by the reaction of ${(cyclo-C_6H_11)}_2Te$ with the corresponding alkyl halide. Reaction of 1 with NaBPh4 gave compound 5 ( i.e. R = CH3, X = BPh4 ‒) in 78% yield. Reaction of ${(cyclo-C_6H_11)}_2Te$ with benzyl bromide and 4-bromophenacyl bromide gave unexpectedly dibenzylcyclohexyltelluronium bromide (6) and bis(4-bromophenacyl)cyclohexyltelluronium bromide (7), respectively. Reaction of 6 with NaBPh4 gave the corresponding tetraphenylborate derivative (8) in high yield. $^1H$ NMR studies revealed that in $CDCl_3$solution compound 1 eliminated alkyl halide. Conductivity, IR, $^1H\;and\;^{13}C$ NMR and thermal data for the new compounds are presented and discussed.

Biological control of the Nothern Root-knot Nematode, Meloidpgyne hapla with Plant Extract (식물의 추출물을 이용한 당근뿌리혹선충의 생물적방제)

  • 김형환;추호렬;박정규;이상명;김준범
    • Korean journal of applied entomology
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    • v.37 no.2
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    • pp.119-206
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    • 1998
  • Nematicidal potential of some plant extracts were evaluated for the control efficacy of the northern root-knot nematode, Meloidogyne hapla on tomato (Lycopersicon esculentum) in pot. Tagetes patula, Zoysia japonica, Rhus sylvestris, R. chinensis, and Allium cepu were used. Leaf or root extracts were prepared at the rate of undiluted and diluted to 2, 4, 8, or 16 times with distilled water and treated simultaneously, ahead or later tomato planting. Pre-treatments of extracts were more effective than simultaneous or post-treatments, and the number of egg masses was different according to concentrations. M. hapla was less infected when the T. patula was planted at 15 days ahead tomato planting. Leaf or root extracts of T. patulu reduced damages of M. hapla significantly in all treatments. Leaf extract was more effective than root extract; the number of egg masses was 2.2 and 5.5 in 5-days pre-treatment of leaf or root extracts while 129.4 in control. In Z. japonica treatment, the number of egg masses was 87.2 in control, 21 in undiluted concentration of leaf extract and 28.4 in diluted concentration as 2 times. Leaf extracts of R. sylvestris, R. chinensis and A. cepa were also very effective against M. Izapla. The number of egg masses of M. hapla was 1.6, 1.6, and 6.2 in 5-days pre-treatments of R. sylvestris, R. chinensis and A. cepa, respectively while it was 193.6 in control. Less egg masses of M. hapla were detected in higher concentrations than in lower concentrations of all the plant extracts.

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Characteristics and Action Pattern of Polygalacturonase from Rhizopus oryzae CJ-2114 (Rhizopus oryzae CJ-2114가 생성하는 Polygalacturonase의 특성 및 작용양상)

  • Chung, Yung-Gun;Cho, Young-Je;Kwon, Oh-Jin;Choi, Cheong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.2
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    • pp.195-200
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    • 1992
  • Rhizopus oryzae CJ-2114 was selected for its strong polygalacturonase activity among various strains of mold found in soil. The optimum pH for the enzyme activity was 4.0 and optimum temperature was 4$0^{\circ}C$. The activation energy for the polygalacturonase was calculated by Arrhenius equation was 2.048㎉/㏖. The reaction of this enzyme followed typical Michaelis-Menten kinetics with the Km value of 54.05mM with the $V_{max}$ of 13.9m mole/min. The enzyme is relatively stable in acidic condition. The activity of polygalactur-onase was inhibited completely by C $u^{2+}$, P $b^{2+}$ and Z $n^{2+}$, $_Mn^{2+}$ at concentration of 1 mM. The enzyme can be inactivated by the treatment with maleic anhydride and iodine. The results indicate the possible involvement of histidine at active site. When polygalacturonase from Rhizopus oryzae CJ-2114 was reacted with poly-galacturonic acid as a substrate mono-, di-, and oligogalacturonic acid were produced at early and mono-, digalacturonic acid produced at late incubation time. time.

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