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Isolation and Purification of Antimicrobial Peptide from Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 유래 항균 펩타이드 분리 및 정제)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;An, Cheul Min;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1259-1268
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    • 2016
  • In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by $C_{18}$ reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], $20.8{\mu}g/ml$), Bacillus subtilis (MEC, $0.2{\mu}g/ml$), Streptococcus mutans (MEC, $0.2{\mu}g/ml$), gram-negative bacteria including Pseudomonas aeruginosa (MEC, $5.7{\mu}g/ml$), Escherichia coli (MEC, $2.6{\mu}g/ml$) and fungi, Candida albicans (MEC, $56.3{\mu}g/ml$). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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The Effects of Protozoa on the Early Formation of Microbial Fouling Communities of Inchon Coastal Waters (인천 연안 미세오손생물 군집 형성과 원생동물의 영향)

  • Choi, Joong-Ki;Yang, Eun-Jin;Lee, Won-Je;Yoon, Won-Duk;Shim, Jae-Hyung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.4 no.4
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    • pp.349-362
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    • 1999
  • To understand the role of protozoa in the early formation of microbial fouling community, the studies on the formation of microbial film, the succession of microbial fouling communities, and the grazing pressure on bacteria population in microbial film were carried out in the laboratory, Inchon outer port and Inchon inner harbour. Bacteria and heterotrophic flagellates formed primary microbial film on the aluminum surface within 6 hours and oligotrich ciliates were observed 2 cells $mm^{-2}$ on the same surface at 9 hours in Inchon inner harbour which had physically stagnant condition. The larvaes of Balanus albicostatus which were dominant meiobenthos in Inchon coastal area attached on the glass surface at the first day of experiment. Heterotrophic flagellates showed maximum abundance of 465 cells $mm^{-2}$ at the 13rd day and ciliates showed maximum abundance of 63 cells $mm^{-2}$ at the 11st day in the Inchon inner harbour. In the Inchon outer port which opens to the outer sea, the maximum abundance of protozoa occurred at early phase, but not so many. The dominant heterotrophic flagellates were Metrornonas simplex and Bodonids. Dominant ciliates were small tintinnids and oligotrich ciliate Strombidium sp., Large Strombidium (oligotrich ciliate) and sessile Acineta turberosa (suctorian ciliate) occurred after 10 days. The attached larvae of Balanus occurred as biofouling organism on the early surface and showed maximum abundance of 18 indiv. $cm^{-2}$ at 7th day. At that time, adult barnacles were observed on the surface and dead barnacles were observed after two days. Except barnacles, the larvaes of Anthozoa sp., Oysters (Crassostrea gigas) and Polychaeta were observed on the surface from 3rd day. 3 benthic copepods including Harpacticus sp., I isopod, 1 polychaeta and 1 gastropoda were observed as predators of the microbial film on the surface after 7 days when microbial film developed very well. Although the ingestion rates of protozoa on the bactctia of the rnicrobi31 film were relatively low, the average grazing rate of protozoa on bacteria was high of 0.058 $h^{-1}$. This implied that the grazing pressure of protozoa influences the mortality of bacteria populations on the microbial film. but protozoa cannot get enough energy from only bacteria on the microbial film.

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A Study on the Amount and Chemical Compositions of Excreta from Laying Hens (산란계의 분뇨 배설량 및 이화학적 성상)

  • HwangBo, Jong;Hong, Eui-Chul;Na, Seung-Hwan;Yu, Dong-Jo;Kim, Hak-Kyu;Park, Mi-Na;Jung, Kie-Chul;Choo, Hyo-Jun;Park, Hee-Du;Seo, Ok-Suk
    • Korean Journal of Poultry Science
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    • v.36 no.4
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    • pp.337-342
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    • 2009
  • This work was conducted to investigate the manure excreta and chemical compositions of layers. Two hundred 1-d-old ISA Brown layers were used in this work, and ten of 200 layers were selected for measurement of manure excreta with feeding phase. Means of the manure excreta per bird, the moisture content of poultry, and the dried manure were $124.7\;{\pm}\;27.5\;g$/bird/day, $76.9\;{\pm}\;1.9%$, and $29.1\;{\pm}\;3.9\;g$/bird/day, respectively. N, $P_2O_5$, and $K_2O$ of manure (55 weeks) were $1.39\;{\pm}\;0.139$, $0.62\;{\pm}\;0.11$ and $0.68\;{\pm}\;0.09%$, respectively. pH, $BOD_5$, $COD_{Mn}$, and suspended solids (SS) were $8.19\;{\pm}\;0.71$, $50,266\;{\pm}\;621\;mg/L$, $62,832\;{\pm}\;803\;mg/L$, and $121,725\;{\pm}\;16,165\;mg/L$, respectively. Heavy metal contents in the manure were $0.003\;{\pm}\;0.0013\;ppm$ for Hg and $0.008\;{\pm}\;0.0012\;ppm$ for Pb, the latter of which was highly low compared with the standard (150 ppm), but no Cd was detected. The results of this work show the average excretion amounts and chemical compositions of layer's manure, which is used as an organic fertilizer.

Australian Case Study in Regulatory Techniques to the Security Industry Reform and Policy Implications (호주 민간경비산업 고품질 규제수단 검토 및 시사점)

  • Kim, Dae-Woon
    • Korean Security Journal
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    • no.47
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    • pp.7-36
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    • 2016
  • The security providers industry, often referred to as an industry with unconfined growth ceiling, has entered a remarkable mass-growth phase since the 1980. In the modern era, private-sector security increasingly cover functions relating to general security awareness (including counter-terrorism) in partnership with State bodies, and the scale of operations continue to accelerate, relative to the expanding roles. In the era of pluralisation of policing, there has been widening efforts pursued to develop a range of regulatory strategies internationally in order to manage such growth and development. To date, in South Korea, a diverse set of industry review studies have been conducted. However, the analyses have been conventionally confined to North America, Britain, Germany and Japan, while developments in other world regions remain unassessed. This article is intended to inform the drivers and determinants of regulatory reforms in Australia, and examine the effectiveness of the main pillars of licensing innovations. Over the past decades, the Australian regime has undergone a wave of reforms in response to emerging issues, and in recognition of the industry as a 'public good' due to underpopulation density and the resulting security challenges. The focus of review in this study was on providing a detailed review of the regulatory approach taken by Australia that has expanded police-private security co-operation since the 1980s. The emphasis was on examining the core pillars of risk management strategies and oversight practices progressed to date and evaluating areas of possible improvement in regulation relative to South Korea. Overall, this study has identified three key features of Australian regime: (1) close checks on questionable close associates (including fingerprinting), (2) power of inspection and seizure without search warrant, (3) the 'three strikes' scheme. The rise of the private security presence in day-to-day policing operations means that industry warrant some intervening government-sponsored initiative. The overall lessons learnt from the Australian case was taken into account in determining the following checks and balances that would provide the ideal setting for the best-practice arrangement: (1) regulatory measure should be evaluated against a set of well-defined indicators, such as the merits of different enforcement tools for each given risk, (2) information about regulatory impacts should be analysed by a specialist research institute, (3) regulators should be innovative in applying a range of strategies available to them by employing a mixture of compliance promotional strategies, and adjust the mix as required.

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Analysis of golf swing motion for specific properties of club shaft (클럽 샤프트(Club Shaft) 특성에 따른 골프 스윙(Golf Swing)동작 분석)

  • Kim, Sung-Il;Kim, Ky-Hyoung;Kim, Hyung-Soo;Lee, Hyun-Seob;Kim, Jin-Uk;Ahn, Chan-Gyu;Kim, Hee-Jin
    • Korean Journal of Applied Biomechanics
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    • v.12 no.2
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    • pp.17-32
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    • 2002
  • The purpose of this study was to find the rational method to analyze golf swing with specific property of club shaft. Three subjects were filmed by two high speed digital cameras with 500 fps. The phase analyzed was downswing of each subject. The three-dimensional coordinates of the anatomical landmarks were obtained with motion analysis system Kwon3d 3.0 version and smoothed by lowpass digital filter with cutoff frequency 6Hz. From these data, kinematic and kinetic variables were calculated using Matlab(ver 5.0) The variables for this study were angular velocity and accelerations, which were calculated and following conclusions have been made : 1) Golf swing time of stiff club is faster than that of regular club. 2) In shoulder joint motion of swing with the stiff club, x-stiff showed mort rapid negative acceleration than that of regular club. 3) In regular club, the velocity of club head would be more effective velocity, which was increasing, than those of other clubs before impact. 4) In wrist joint motion of swing with stiff club, x-stiff club showed faster than regular club in the downswing and impact more rapid negative acceleration.

A Case Study on Center of Gravity Analysis when Performing Uchimata by Posture and Voluntary Resistance Levels of Uke in Judo[ll] (유도 허벅다리걸기 기술발휘 시 받기의 자세와 저항수준에 따른 중심변인 분석 사례연구[II])

  • Kim, Eui-Hwan;Kim, Sung-Sup;Chung, Chae-Wook
    • Korean Journal of Applied Biomechanics
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    • v.15 no.1
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    • pp.237-257
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    • 2005
  • It was to study as a following-research of "A Case Study on Center of Gravity(COG) Analysis when Performing Uchimata(inner thigh reaping throw) by Posture and Voluntary Resistance Levels(VRL) of Uke in Judo[I]". The purpose of this study was to analyze the COG variables when performing uchimata(inner thigh reaping throw) by two postures and voluntary resistance levels(VRL) of uke(reciver) in Judo. The subjects, who were one male judoka(YH) for 1992 Barcelona Olympic Games Olympian(silver medalist), and one male trainee; Y.I.University representative member (SDK), and were filmed on two S-VHS 16mm video cameras(60fields/sec.) through 3-dimensional motion analysis methods, that postures of uke were shizenhontai (straight natural posture) and jigohontai(straight defensive posture), VRL of uke were 0% and 100%, respectively. The kinematical variable was COG variable, distance of COG, and distance of resultant COG between uke and tori(the thrower), velocity and acceleration of COG. The data of this study collection were digitized by SIMI Motion Program computed the mean values and the standard deviation calculated for each variables. When performing uchinmata according to each posture and VRL of uke and classifying. From the data analysis and discussion, the conclusions were as follows : 1. Displacement of COG Subject YH, COG was the highest in kuzushi(balance -breaking), vertical COG was low when following in tsukuri(positioning; set-up), kake(application; execution), and COG was pattern of same character each postures and resistance, respectively. Subject SDK, COG was low from kumikata(engagement positioning) to kake, and COG was that each postures and resistance were same patterns, respectively. Subject YH, SDK, each individual, postures and resistance, vertical COG was the lowest in kake phase, when performing. 2. Distance of COG between uke and tori The distance of COG between uke and tori when performing, subject YH was $0.64{\sim}0.70cm$ in kumikata, $0.19{\sim}0.28cm$ in kake, and SDK was $0.68{\sim}0.72cm$ in kumikata, $0.30{\sim}0.42\;cm$ in kake. SDK was wider than YH. 3. Distance of resultant COG between uke and tori The distance of resultant COG between uke and tori when performing, subject YH was $0.27{\sim}0.73cm$ from kumikata to kake. and SDK was $0.14{\sim}0.34cm$ in kumikata, $0.28{\sim}0.65cm$ in kake. Jigohontai(YH:$0.43{\sim}0.73cm$,SDK:$0.59{\sim}0.65cm$) was more moved than shizenhontai(YH:$0.27{\sim}0.53cm$, SDK: $0.28{\sim}\;0.34cm$). 4. Velocity of COG The velocity of COG when performing uchimata, subject YH was fast anterior-posterior direction in kuzushi, ant.-post. and vertical direction fast in tsukuri and kake. SDK was lateral, ant.-post. and vertical direction in kuzushi, ant.-post. and vertical direction in tsukuri and ant.-post. direction in take, respectively. 5. Acceleration of COG The acceleration of COG when performing uchimata, The trend of subject YH was showed fast vertical direction in kuzushi and tsukuri, ant.-post. and vertical direction fast in kake. The trends of SDK showed lateral direction in kuzushi, lateral and ant.-post. direction in tsukuri and ant.-post. direction in kake, respectively.

Effectiveness of Fentanyl Transdermal Patch (Fentanyl-TTS, $Durogegic^{(R)}$) for Radiotherapy Induced Pain and Cancer Pain: Multi-center Trial (방사선치료로 인한 통증 및 암성통증에 대한 듀로제식의 효과: 다기관연구)

  • Shin, Seong-Soo;Choi, Eun-Kyung;Kim, Jong-Hoon;Ahn, Seung-Do;Lee, Sang-Wook;Kim, Yeun-Sil;Lee, Kyu-Chan;Lee, Chang-Geol;Loh, John-JK;Chun, Mi-Son;Oh, Young-Teak;Kim, Ok-Bae;Huh, Seung-Jae
    • Radiation Oncology Journal
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    • v.24 no.4
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    • pp.263-271
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    • 2006
  • $\underline{Purpose}$: To evaluate the effectiveness and safety of fentanyl-TTS in the management of radiotherapy induced acute pain and cancer pain treated with radiotherapy. $\underline{Materials\;and\;Methods}$: Our study was open labelled prospective phase IV multi-center study. the study population included patients with more 4 numeric rating scale(NRS) score pain although managed with other analgesics or more than 6 NRS score pain without analgesics. Patients divided into two groups; patients with radiotherapy induced pain (Group A) and patients with cancer pain treated with radiotherapy (Group B). All patients received 25 ug/hr of fentanyl transdermal patch. Primary end point was pain relief; second end points were change in patient quality of life, a degree of satisfaction for patients and clinician, side effects. $\underline{Results}$: Between March 2005 and June 2005, 312 patients from 26 participating institutes were registered, but 249 patients completed this study. Total number of patients in each group was 185 in Group A, 64 in Group B. Mean age was 60 years and male to female ratio was 76:24. Severe pain NRS score at 2 weeks after the application of fentanyl was decreased from 7.03 to 4.01, p=0.003. There was a significant improvement in insomnia, social functioning, and quality of life. A degree of satisfaction for patients and clinician was very high. The most common reasons of patients' satisfactions was good pain control. Ninety six patients reported side effect. Nausea was the most common side effect. There was no serious side effect. $\underline{Conclusion}$: Fentanyl-TTS was effective in both relieving pain with good tolerability and improving the quality of life for patients with radiotherapy induced acute pain and cancer pain treated with radiotherapy. The satisfaction of the patients and doctors was good. There was no major side effect.

Cytotoxicity of COX-2 Inhibitor (Nimesulide) in Non-small Cell Lung Cancer Cell Line (비소세포폐암 세포주에서 COX-2억제제(Nimesulide)의 세포독성)

  • Park Chan Beom;Jeon Hyun Woo;Jin Ung;Cho Kyu Do;Kim Chi Kyung;Wang Young-Pil
    • Journal of Chest Surgery
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    • v.38 no.4 s.249
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    • pp.263-270
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    • 2005
  • In recent years, a combination of two demographic phenomena, an increased number of older people in the population and an increase in the incidence of lung cancer with age, has made it mandatory to develop therapeutic modalities with less toxicity for the treatment of inoperable elderly patients with lung cancer. Therefore, we investigated the correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Material and Method: Immunohistochemical staining of COX-2 was performed. After exposure of Nimesulide, XTT analysis, FACS analysis and Hoechst staining were carried out. Result: COX-2 protein was expressed in non-treated A549 cells strongly, but not in H1299. Cytotoxicity of Nimesulide against A549 cell and H1299 cell were similar and $IC_{50}$ of Nimesulide in both cell lines were $70.9{\mu}M$ in A549 cell line and $56.5{\mu}M$ in H1299 cell line respectively. FACS analysis showed $G_0/G_1$ arrest in both cell lines and the S phase cell fraction was decreased. Morphologic assessment of apoptosis by Hoechst 33258 staining, many apoptotic cells were detected in both cell lines. Conclusion: Selective COX-2 inhibitor, Nimesulide, can inhibit the proliferation of non-small cell lung cancer cell lines in vitro. Inhibitory effect of Nimesulide are induction of apoptosis and $G_0/G_1$ arrest. There is no correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Therefore, highly selective COX-2 inhibitors such as Nimesulide can be expected to lead to even greater efficacy of their use as adjuncts to various anticancer angents and radiation therapy for the treatment of high-risk patients.

Resolution Method of Hazard Factor for Life Safety in Rental Housing Complex (임대주택단지의 생활안전 위해요인 해소방안)

  • Sohn, Jeong-Rak;Cho, Gun-Hee;Kim, Jin-Won;Song, Sang-Hoon
    • Land and Housing Review
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    • v.8 no.1
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    • pp.1-11
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    • 2017
  • The government has been constructing and supplying public rental housing to ordinary people in order to stabilize housing since 1989. However, the public rental houses initially supplied to ordinary people are at high risk for safety accidents due to the deterioration of the facilities. Therefore, this study is aimed to propose a solution to solve the life safety hazards of the old rental housing complex as a follow-up study of Analysis of Accident Patterns and Hazard Factor for Life Safety in Rental Housing Complex. Types of life safety accidents that occur in public rental housing complexes are sliding, falling, crash, falling objects, breakage, fire accidents, traffic accidents and criminal accidents. The types of safety accidents that occur in rental housing complexes analyzed in this study are sliding, crashes, falling objects, and fire accidents. Although the incidence of safety accidents such as falling, breakage, traffic accidents and crime accidents in public rental housing complexes is low, these types are likely to cause safety accidents. The method of this study utilized interviews and seminar results, and it suggested ways to solve the life safety hazards in rental housing complexes. Interviews were conducted with residents and managers of rental housing complexes. Seminars were conducted twice with experts in construction, maintenance, asset management, housing welfare and safety. Through interviews and seminars, this study categorizes the life safety hazards that occur in rental housing complexes by types of accidents and suggests ways to resolve them as follows. (1) sliding ; use of flooring materials with high friction coefficient, installation of safety devices such as safety handles, implementation of maintenance, safety inspections and safety education, etc. (2) falling ; supplementation of safety facilities, Improvement of the design method of the falling parts, Safety education, etc. (3) crash ; increase the effective width of the elevator door, increase the effective width of the lamp, improve the lamp type (U type ${\rightarrow}$ I type), etc. (4) falling objects and breakage ; design of furniture considering the usability of residents, replacement of old facilities, enhancement of safety consciousness of residents, safety education, etc. (5) fire accidents ; installation of fire safety equipment, improvement by emergency evacuation, safety inspection and safety education, etc. (6) traffic accidents ; securing parking spaces, installing safety facilities, conducting safety education, etc. (7) criminal accidents; improvement of CCTV pixels, installation of street lights, removal of blind spots in the complex, securing of security, etc. The roles of suppliers, administrators and users of public rental housing proposed in this study are summarized as follows. Suppliers of rental housing should take into consideration the risk factors that may arise not only in the design and construction but also in the maintenance phase and should consider the possibility of easily repairing old facilities considering the life cycle of rental housing. Next, Administrators of rental housing should consider the safety of the users of the rental housing, conduct safety checks from time to time, and immediately remove any hazardous elements within the apartment complex. Finally, the users of the rental housing needs to form a sense of ownership of all the facilities in the rental housing complex, and efforts should be made not to cause safety accidents caused by the user's carelessness. The results of this study can provide the necessary information to enable residents of rental housing complexes to live a safe and comfortable residential life. It is also expected that this information will be used to reduce the incidence of safety accidents in rental housing complexes.