• Title/Summary/Keyword: $E_b/N_t$

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Overexpression and Periplasmic Transport of 5-Enolpyruvylshikimate 3-Phosphate Synthase in E. coli (대장균에서 5-Enolpyruvylshikimate 3-Phosphate Synthase의 대량 발현 및 Periplasmic Space로의 Transport)

  • 김남일;임재윤;조태주
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.1-6
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    • 1997
  • 5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase is the sixth enzyme of the shikimate pathway that synthesizes aromatic amino acids. The enzyme is a primary target for the glyphos'lte which is a broad-spectrum and environmetally safe herbicide. As a first step toward development of glyphpsate-resistant EPSP synthase, the EPSP synthase gene(aroA) was amplified by polymerase chain reaction and cluned into pET-25b vector. In this construct. designated pET-aro, the aroA gene is expressed under control of strong T7 promoter. and the EPSP synthase is produced as a fusion protein with pelB leader at N-terminus and HSV-tag and His-tag at C-terminus. When the pET-aro clone was induced to produce the enzyme, it was found that the EPSP synthase was successfully exported to peri plasmic space. The periplasmic transport was greatly dependent on the induction temperatures. Among the induction temperatures examined($25^{\circ}C$, $30^{\circ}C$, $34^{\circ}C$ and $37^{\circ}C$). induction at $34^{\circ}C$ gave rise to maximal periplasmic transport. The recomhinant EPSP synthase could have been purified hy $Ni^{2+}$ -affinity chromatography using the His-tag. and detected hy anti-HSV -tag antibody. The recombinant EPSP synthase also hound to phosphocellulose resin and was eluted hy shikimate 3-phosphate and phosphoenolpyruvate. as expected. The recombinant EPSP synthase purified from phosphocellulose resin showed typical EPSP synthase activity.

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Effects of Wilting and Additives on the Fermentation Chrateristics , Quality and Aerobic Stability of Rye Silage (예건 및 첨가제가 호밀사일리지의 발효특성과 사료가치 및 호기적 안정성에 미치는 영향)

  • 이광녕;김동암
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.17 no.2
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    • pp.187-198
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    • 1997
  • This experiment was conducted to evaluate effects of wilting and additives on fermetation characteristics, quality and aerobic stability of silage of rye(Seca1e cereale L.) harvested at the early heading stage. Harvested rye was wilted or mixed with beet pulp and then treated with formic acid or lactic acid bacteria(LAB) inoculant. Treatments were consisted of direct cut(DC), wilted(WT), beet pulp added(BP), direct cut and formate applied(DF), wilted and formate applied(WF), beet pulp and formate applied(BF), direct cut and LAB inoculated(DL), beet pulp added and LAB inoculated(BL), and wilted and LAB inoculated(WL) silages. Afler 70 days of ensiling period, the silages were opened and exposed to air for 7 days, and the silages of opening day(0 day) were compared with the silages exposed to air for 7 days(7 day). 1. LAB inoculated rye silages(DL, WL, BL) lowered pH values effectively eom the third day of ensiling and showed most stable pattern of pH changes during the initial fermentation process. Direct cut and formate applied(DF), beet pulp and formate applied(BF) and beet pulp added(BP) silages were also effective in lowering pH from the seventh day thai direct cut treatment(DC). 2. pH was low below 3.7 in all treatments of 0 day. After 7-day period of aerobic exposure, DL and WL showed considerable increases in pH from 3.5 and 3.4 to 8.3 and 6.4, respectively. 3. Direct cut rye silages(DF, DC, DL) and beet pulp and formate treatment(BF) produced effluent of 121.2, 85.9, 80.3 and 34.2 mlkg, respectively and these were greater than others(P<0.01). Beet pulp retained 1.61 I/kg of effluent at the application rate of 50 kg/t in comparing BP with DC, and formate application increased the amount of effluent in comparing DF and BF with DC and BP, respectively(P<0.01). 4. Beet pulp and LAB treatment(B1) was highest in content of lactate as 3.1% and formate treated silages(DF, WF, BF) showed lower content of lactate and total acid than others. LAB inoculated(D1, WL, BL) and wilted (WT) silages were graded to be good quality as a second group by the Flieg's score. Formate applied silages (DF, WF, BF) were graded as a foum group lower than DC by one. 5. Residual content of WSC of rye silage was higher in BP, BF, DL and BL than direct cut treatment(M3) (P< 0.01). Formate application had a tendency to increase the content of residual content of WSC. 6. For the ratio of NH3-N to Total N, wilted and LAB inoculated(W1) and direct cut and LAB inoculated(D1) silages were lowest on 0 day as 8.9 and 9.3% respectively. But after 7-day period of aerobic exposure, WL and DL showed largest increase of the ratio of $NH_3-N$ to Total N from 0 day(P<0.01). 7. On 0 day wilted(WT), beet pulp and formate treatment(BF), and beet pulp and LAB treatment(B1) were lower than direct cut treatment(DC) for NDF and ADF concentrations consistently. Formate applied silages (DF, WF, BF) made little change in ADF and NDF concentrations during 7 day period of aerobic exposure. DL and WL showed a large increase in ADF and NDF concentrations under aerobic condition(P<0.01). 8. IVDMD values of wilted(WT), beet pulp added(BP), wilted and LAB inoculated(W1) and beet pulp added and LAB inoculated(B1) silages were higher than direct-cut treatmentPC) as 84.7, 84.7, 84.4 and 83.0%, respectively on 0 day. But during 7-day period of aerobic exposure, a great decreae in IVDMD of WL was showed(P<0.01). The experimental results indicate that wilted silage(WT) could be recommended as the most effective treatment for reducing efluent and increasing quality and feed value of rye silage without deteriorating aerobic stability more than direct cut treatment(DC). Additionally, under unfavorable weather condition beet pulp added and LAB inoculated treatment(T3L) might be the possible alternative for successful ensiling of forage rye.

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

The Study to Re-establish the Amount and Major Pollution Source of Wastewater from Seasonal Swine Farms (양돈농가에서의 계절별 세정수 발생량 및 주요 오염원 재설정 연구)

  • Kwag, J.H.;Choi, D.Y.;Kim, J.H.;Jeong, K.H.;Cho, S.H.;Jeong, M.S.;Kang, H.S.;Ra, C.S.
    • Journal of Animal Environmental Science
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    • v.15 no.3
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    • pp.225-230
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    • 2009
  • The purpose of this study was to determine the effect of re-establish the amount and major pollution source of swine wastewater from swine farms. The results obtained in this study was summarized as follow; The quantity of wastewater produced from the average volume of swine wastewater was $2.49{\ell}$/head/day and $2.49{\ell}$ in spring, $2.65{\ell}$ in summer, $2.47{\ell}$ in autumn, and $2.35{\ell}$ in winter. The quantity of wastewater production during summer season was higher than of the other seasons. The water pollutant concentration in wastewater of swine farms, $BOD_5$, SS, T-N and T-P, was $4,689\;mg/{\ell}$, $2,122\;mg/{\ell}$, $3,238\;mg/{\ell}$ and $118\;mg/{\ell}$, respectively.

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Analysis of Vegetation Structures and Vegetation-Environment Relationships of Medicinal on Short-term Income Forest Products, in Korea - Cudrania tricuspidata (Carrière) Bureau ex Lavallèe·Sorbus commixta Hedl.·Hovenia dulcis Thunb. - (임산물 약용수의 자생지 식생 구조와 환경과의 상관관계 분석 - 꾸지뽕나무·마가목·헛개나무 -)

  • Hyoun-Sook Kim;Sang-Myong Lee;Kil-Nam Kang;Seog-Gu Son;Si-Chul Ryu;Kyung-Joon Lee;Jong-Hoon Lee;Byung-Seol Lee;Joong-Ku Lee
    • Korean Journal of Environment and Ecology
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    • v.37 no.5
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    • pp.347-366
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    • 2023
  • In the present study, the vegetation was classified using the phytosociological method and canonical-correlation analysis (CCA) was implemented to analyze correlation between community structure and environmental factors in the natural habitats of forest byproducts, especially medicinal plants, such as Cudrania tricuspidata, Sorbus commixta, and Hovenia dulcis, in 2021-2022 to provide primary ecological data to establish environmental conditions for wild vegetable cultivation. A total of 11 plots in five regions, 8 plots in three regions, and 17 plots in 5 regions were selected for the natural habitats of C. tricuspidata in southern Korea, S. commixta in high mountains, and H. dulcis in valleys of central Korea, respectively. The importance value in each community was respectively analyzed as follows, in C. tricuspidata community, the importance value of C. tricuspidata (61.10) was the highest, followed by Celtis sinensis, Pinus thunbergii, Neolitsea aciculata, Styrax japonica, Carpinus coreana, Quercus serrata, and Q. acutissima. In Sorbus commixta community, Q. mongolica (57.21) was the highest, followed by, S. commixta (42.58), Betula ermani, Tilia amurensis, A. pseudosieboldianum, A. tschonoskii var. rubripes, Cornus controversa, Magnolia sieboldii, and Taxus cuspidata. In H. dulcis community, H. dulcis (64.58) was the highest, followed by Zelkova serrata, Cornus controversa, A. mono, Q. serrata, C. cordata, and Juglans mandshurica. As the result of the analysis on DBH of the major species having the high importance value, in C. tricuspidata community, C. tricuspidata, C. sinensis, Neolitsea aciculata, and C. coreana show the density of normal distribution, so the dominant status of these species is likely to continue. In S. commixta community, S. commixta show the density of reverse J-shaped curve, so the dominant status of these species is likely to be stable, and Q. mongolica, B. ermani and T. amurensis, show the density of normal distribution, so the dominant status of these species is likely to continue. In H. dulcis community, C. cordata, and J. mandshurica show the density of reverse J-shaped curve, so the dominant status of these species is likely to be stable, and H. dulcis, Z. serrata, C. controversa and A. mono had a formality distribution, suggesting a continuous domination of these species over the other species for the time being. The results of CCA ordination analysis using 11 environmental factors and 30 communities of three taxa classified by TWINSPAN analysis revealed that the altitude showed the strongest correlation with the vegetation. C. tricuspidata community was distributed on the moderate and gentle northeastern slope at low altitude with the highest pH, C.E.C, Ca2+, and Mg2 and various P2O5, whereas S. commixta community was distributed on the steep slope at high altitude with the highest O.M and T-N and lower P2O5, Ca2+, Mg2+, C.E.C and pH, which is the opposite tendency of the environment of C. tricuspidata community. H. dulcis community was distributed on the gentle northern slope at lower altitude with an average pH, O.M, T-N, Ca2+, Mg2+, and C.E.C, except higher P2O5.

Chito-oligosaccharides as an Alternative to Antimicrobials in Improving Performance, Digestibility and Microbial Ecology of the Gut in Weanling Pigs

  • Han, K.N.;Kwon, I.K.;Lohakare, J.D.;Heo, S.;Chae, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.556-562
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    • 2007
  • A total of 126 crossbred weanling pigs (average body weight of $6.3{\pm}0.3$ kg) were used to investigate the effect of chito-oligosaccharide (COS) on growth performance, nutrient digestibility, pH of gastro-intestinal tract (GI), intestinal and fecal microflora of young piglets. Pigs were allocated to three dietary treatments based on body weight and gender in a single factorial arrangement. Treatments were control (No COS), T1 (0.2% COS during starter (6-13 kg) and 0.1% COS during grower (13-30 kg) phases, and T2 (0.4% COS during starter (6-13 kg) and 0.3% COS during grower (13-30 kg) phases, respectively. Each treatment had 3 replicates and 14 pigs were raised in each pen. COS is a low molecular weight water-soluble chitosan that can be obtained from chitin of the crab shell after deacetylation with concentrated sodium hydroxide at high temperature and then further decomposition by chitosanase enzyme in the presence of ascorbic acid. For the starter and grower periods, there were no significant differences (p>0.05) in average daily gain (ADG) and feed to gain ratio among treatments. However, during the overall period (6-30 kg), T2 showed better (p<0.05) feed to gain ratio than other treatments. A digestibility study was conducted at the end of grower phase which showed improvement (p<0.05) in DM and crude fat digestibility in T2 over the control. At 25 kg body weight, 6 pigs per treatment (2 per replicate) were sacrificed to determine the effect of diets on pH and microbial count at different sections of the GI tract. The pH of the cecal contents in pigs fed 0.1% COS was higher (p<0.05) than in the other treatments. Total anaerobic bacterial number increased from cecum to rectum in all treatments. The weekly total bacterial counts showed higher (p<0.05) in feces of pigs fed COS than that of untreated pigs at the $8^{th}$ week. The number of fecal E. coli in untreated pigs at $4^{th}$ wk was 7.35 log CFU/g compared to 6.71 and 6.54 log CFU/g in 0.1 and 0.3% COS-treated pigs, respectively. Similarly, at $8^{th}$ wk, fecal clostridium spp. were lower in pigs fed 0.3% COS (5.43 log CFU/g) than in untreated pigs (6.26 log CFU/g). In conclusion, these results indicated that chito-oligosaccharide could improve feed efficiency in young pigs and inhibited the growth of harmful bacteria.

Mitochondrial Cytochrome b Sequence Variations and Population Structure of Siberian Chipmunk (Tamias sibiricus) in Northeastern Asia and Population Substructure in South Korea

  • Lee, Mu-Yeong;Lissovsky, Andrey A.;Park, Sun-Kyung;Obolenskaya, Ekaterina V.;Dokuchaev, Nikolay E.;Zhang, Ya-Ping;Yu, Li;Kim, Young-Jun;Voloshina, Inna;Myslenkov, Alexander;Choi, Tae-Young;Min, Mi-Sook;Lee, Hang
    • Molecules and Cells
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    • v.26 no.6
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    • pp.566-575
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    • 2008
  • Twenty-five chipmunk species occur in the world, of which only the Siberian chipmunk, Tamias sibiricus, inhabits Asia. To investigate mitochondrial cytochrome b sequence variations and population structure of the Siberian chipmunk in northeastern Asia, we examined mitochondrial cytochrome b sequences (1140 bp) from 3 countries. Analyses of 41 individuals from South Korea and 33 individuals from Russia and northeast China resulted in 37 haplotypes and 27 haplotypes, respectively. There were no shared haplotypes between South Korea and Russia - northeast China. Phylogenetic trees and network analysis showed 2 major maternal lineages for haplotypes, referred to as the S and R lineages. Haplotype grouping in each cluster was nearly coincident with its geographic affinity. In particular, 3 distinct groups were found that mostly clustered in the northern, central and southern parts of South Korea. Nucleotide diversity of the S lineage was twice that of lineage R. The divergence between S and R lineages was estimated to be 2.98-0.98 Myr. During the ice age, there may have been at least 2 refuges in South Korea and Russia - northeast China. The sequence variation between the S and R lineages was 11.3% (K2P), which is indicative of specific recognition in rodents. These results suggest that T. sibiricus from South Korea could be considered a separate species. However, additional information, such as details of distribution, nuclear genes data or morphology, is required to strengthen this hypothesis.

Effect of Lipid Sources with Different Fatty Acid Profiles on Intake, Nutrient Digestion and Ruminal Fermentation of Feedlot Nellore Steers

  • Fiorentini, Giovani;Carvalho, Isabela P.C.;Messana, Juliana D.;Canesin, Roberta C.;Castagnino, Pablo S.;Lage, Josiane F.;Arcuri, Pedro B.;Berchielli, Telma T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1583-1591
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    • 2015
  • The present study was conducted to determine the effect of lipid sources with different fatty acid profiles on nutrient digestion and ruminal fermentation. Ten rumen and duodenal fistulated Nellore steers (268 body weight${\pm}27kg$) were distributed in a duplicated $5{\times}5$ Latin square. Dietary treatments were as follows: without fat (WF), palm oil (PO), linseed oil (LO), protected fat (PF; Lactoplus), and whole soybeans (WS). The roughage feed was corn silage (600 g/kg on a dry matter [DM] basis) plus concentrate (400 g/kg on a DM basis). The higher intake of DM and organic matter (OM) (p<0.001) was found in animals on the diet with PF and WF (around 4.38 and 4.20 kg/d, respectively). Treatments with PO and LO decreased by around 10% the total digestibility of DM and OM (p<0.05). The addition of LO decreased by around 22.3% the neutral detergent fiber digestibility (p = 0.047) compared with other diets. The higher microbial protein synthesis was found in animals on the diet with LO and WS (33 g N/kg OM apparently digested in the rumen; p = 0.040). The highest C18:0 and linolenic acid intakes occurred in animals fed LO (p<0.001), and the highest intake of oleic (p = 0.002) and C16 acids (p = 0.022) occurred with the diets with LO and PF. Diet with PF decreased biohydrogenation extent (p = 0.05) of C18:1 n9,c, C18:2 n6,c, and total unsaturated fatty acids (UFA; around 20%, 7%, and 13%, respectively). The diet with PF and WF increased the concentration of $NH_3-N$ (p<0.001); however, the diet did not change volatile fatty acids (p>0.05), such as the molar percentage of acetate, propionate, butyrate and the acetate:propionate ratio. Treatments PO, LO and with WS decreased by around 50% the concentration of protozoa (p<0.001). Diets with some type of protection (PF and WS) decreased the effects of lipid on ruminal fermentation and presented similar outflow of benefit UFA as LO.

Microgrooves on titanium surface affect peri-implant cell adhesion and soft tissue sealing; an in vitro and in vivo study

  • Lee, Hyo-Jung;Lee, Jaden;Lee, Jung-Tae;Hong, Ji-Soo;Lim, Bum-Soon;Park, Hee-Jung;Kim, Young-Kwang;Kim, Tae-Il
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.120-126
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    • 2015
  • Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

The Relationship between the Personality of Elementary School Students and the Safety Accidents in the School (초등학생의 성격특성과 학교안전사고)

  • Kim, Chang-Hee;Park, Young-Soo
    • The Journal of Korean Society for School & Community Health Education
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    • v.4
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    • pp.1-20
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    • 2003
  • This study was not intended to analyze out the realities of the safety accidents in school. The purpose of this study was to find out the personality of elementary school students in 6th grade and analyze the relationship between the personality of elementary school students and the safety accidents in the school. We made a visit 12 elementary school personally in the B city of Gyeonggi Province and gave out 552 questionaries in order to accomplish the purpose of this study. And we analyzed the 501 answer data except the unfittable data 51 sheets-in case of no answer or untruthfulness. The inspection of the personality used the Eysenck Personality Questionnaire(EPQ) and the questionnaires of the safety accidents in the school were reorganized of the questionnaires which were used in Chun-Ok, Joen(1997) and One-Yong, Kim(1999)'s study to correspond with our purpose of study. Answer sheets were gone through the t and x2 test by using the program of SPPS. First, elementary school students are in the formative period in the life of a man. So many students's personality was in the middle group rather than in high and low group. In regard of the distintion between the boy students and the girl students, the boy students had higher score in Psychoticism(P) but the girl students had higher score in Neutroticism(N). And Lie(L) was showing a little more score in the girl students. Second, there was significant difference in the investigation of safety accidents in school according to sex distinction. 1) Comparing the boy students and the girl students, the rate of accidents was higher in boy rather than girl. And there was significant difference. 2) Comparing the grade of the school, the rate of accidents was higher in upper grade than lower grade both girls and boys. 3) In the kind of accidents, the traumatic Injuries were very popular and the next were contusions. There was significant difference in the traumatic injury and nosebleed. 4) The results of injured parts showed that the injuries of foot, leg, hand, arm hold a large majority. And there was a significant difference in head, face, foot, leg and etc. 5) Most accidents broke out during the break time and then accidents happened after school were the next. The flowings were lunch time and class time. There was a significant difference in accidents happened during the class, break time and lunch time. 6) Comparing the subjects, the most accidents broke out in the class of physics. 7) Among the cause of accidents, one's own carelessness was showing the highest rate. There was a significant distintion in the fight and game. 8) The safety accidents took place most in playground, and there was a significant difference. 9) Being slight wound was showing almost in the degrees of damage. And there was a significant difference. Third, the boy students were not showing the significant difference in relation to the personality, the frequency of accidents and the degrees of damage. But the girl students were showing the significant difference in relation to the Extraversion-Introversion(E) and the frequency of accidents. And there were a significant difference between E and N. Also they were showing the intense damages.

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