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Comparision of PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS) in Leu72Met Polymorphism of Ghrelin Gene (Ghrelin 유전자의 Leu72Met 다형성 분석에서 PCR-RFLP, PCR-SSCP, Amplication Refractory Mutation System(ARMS)의 비교분석)

  • Kang, Ju Sung;Kim, Se Rim;Kim, Sun Young;Joo, Chan Uhng;Cho, Soo Chul;Hwang, Pyoung Han
    • Clinical and Experimental Pediatrics
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    • v.48 no.10
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    • pp.1068-1075
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    • 2005
  • Purpose : The role of ghrelin, which promotes the secretion of growth hormone, was not well known until now. Recently it was found that the mutation of ghrelin gene is related to obesity and diabetes. This study is to find the screening method that can easily and effectively detect the polymorphism of Leu72Met in ghrelin gene of obesity patients and apply it to clinical usage. Methods : We compared PCR-RFLP, PCR-SSCP and ARMS methodologies for analyzing of the polymorphism of Leu72Met in ghrelin gene of obesity children, and also studied the merits and demerits of these methodologies. Results : In this study, we were able to find out the band of peculiar allele of Leu72Met in ghrelin gene using PCR-RFLP, PCR-SSCP and ARMS analyses. The polymorphism of Leu72Met in ghrelin gene determined by all above methodologies was in complete agreement. Compared to the PCR-RFLP and PCR-SSCP, ARMS analysis is simple, inexpensive and also consume less time. It is very sensitive to analyze the polymorphism and easy to understand the results of test. Conclusion : Though PCR-RFLP, PCR-SSCP and ARMS analyses were sensitive to analyze the polymorphism of Leu72Met in ghrelin gene, ARMS analysis appears to be more efficient than PCR-RFLP and PCR-SSCP. Therefore, we conclude that ARMS analysis is suitable to analyze the polymorphism of Leu72Met in ghrelin gene for large quantity of specimens.

Diagnostic classification and clinical aspects of floppy infants in the neonatal and pediatric intensive care units (신생아 및 소아 중환자실에 입원한 늘어지는 영아(floppy infant)의 진단적 분류 및 임상적 고찰)

  • Kim, Eun Sun;Jung, Kyung Eun;Kim, Sang Duk;Kim, Eo Kyung;Chae, Jong Hee;Kim, Han Suk;Park, June Dong;Kim, Ki Joong;Kim, Beyong Il;Hwang, Yong Seung;Choi Jung-Hwan
    • Clinical and Experimental Pediatrics
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    • v.49 no.11
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    • pp.1158-1166
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    • 2006
  • Purpose : The purpose of this study is to make a diagnostic classification and discuss a diagnostic strategy of floppy infants by investigating clinical, neurological, electrophysiological, and genetic analysis of infants admitted to intensive care units with the complaint of hypotonia. Methods : A retrospective study was performed from Jan. 1993 to Dec. 2005 in neonatal and pediatric intensive care units of Seoul National University Children's Hospital. Clinical features and all tests related to hypotonia were investigated. Results : There were 21 cases of floppy infants admitted to intensive care units. Final diagnosis was classified as centra (7 cases[33.3 percent]), peripheral (11 cases [52.4 percent]), and unspecified (3 cases [14.3 percent]). Among the central group, three patients were diagnosed as hypoxic ischemic encephalopathy, two patients as Prader-Willi syndrome, one patient as chromosomal disorder, and one patient as transient hypotonia. Among the peripheral group, four patients were diagnosed as myotubular myopathy, three patients as SMA type 1, two patients as congenital myotonic dystrophy, one patient as congenital muscular dystrophy, and one as unspecified motor-neuron disease. Motor power was above grade 3 on average, and deep tendon reflex was brisk in the central group. Among investigations, electromyography showed 66 percent sensitivity in the peripheral group, and muscle biopsy was all diagnostic in the peripheral group. Brain image was diagnostic in the central group, and Prader-Willi FISH or karyotyping was helpful in diagnosis in central group. Morbidity and mortality was more severe in the peripheral group Conclusion : Classification of diagnosis by clinical characteristics in this study, and application of investigations step by step, may provide an effective diagnostic strategy.

Allele Distribution and Frequency of Human Surfactant Protein-A2 in Korean Neonates (한국 신생아의 폐 표면 활성제 단백-A2(Human Surfactant Protein-A2) 유전자 대립형질의 분포와 빈도)

  • Kim, Nyeon Cheon;Yoon, Hee Chul;Suk, Jung Su;Ko, Jung Ho;Yoo, Ook Joon;Lee, In Kyu;Oh, Myung Ho;Bae, Chong Woo
    • Clinical and Experimental Pediatrics
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    • v.46 no.4
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    • pp.340-344
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    • 2003
  • Purpose : We evaluated allele frequencies and distribution of surfactant protein A2(SP-A2) in Korean neonates in order to estimate the prevalence of RDS, to find out new SP-A alleles, and to establish new steroid therapy. Methods : Genomic DNA was extracted from 71 neonates and served as a template in PCR for genotype analysis. SP-A gene-specific amplications and gene-specific allele determinations were performed using PCR-cRFLP methods. Results : The distribution for the alleles of the SP-A2 gene in the study population was 1A, $1A^0$, $1A^1$, $1A^2$, $1A^3$, $1A^5$, $1A^6$, $1A^7$, $1A^8$, $1A^9$, $1A^{11}$, $1A^{12}$. The specific frequencies for the alleles of the SP-A2 gene in the study population were : 1A=11.3%, $1A^0=38%$, $1A^1=12.7%$, $1A^2=9.2%$, $1A^5=15.5%$, $1A^7=2.9%$, $1A^8=4.9%$, $1A^9=2.2%$, others=3.3%. Conclusion : The frequency of $1A^0$ was higher than the other SP-A2 alleles in Korean neonates. This finding suggests that the prevalence of RDS in Korea may be low compared with other countries. However, this finding also suggests that Korean neonates have a high risk of infection.

Metabolic syndrome in the overweight and obese adolescents and the impact of obesity on the cardiovascular system (과체중과 비만 청소년에서 대사증후군과 비만이 심혈관에 미치는 영향)

  • Hong, Young Mi;Song, Young Whan;Kim, Hae Soon;Park, Hae Sook;Min, Jung Hae;Jung, Jo Won;Kim, Nam Su;Noh, Chung Il
    • Clinical and Experimental Pediatrics
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    • v.52 no.10
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    • pp.1109-1118
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    • 2009
  • Purpose : Metabolic syndrome (MS), characterized by obesity and insulin resistance, elicits risk factors such as hyperlipidemia, hypertension, and glucose intolerance with additive effects on atherosclerosis, leading to cardiovascular diseases. The purposes of this study were to evaluate the prevalence of MS among overweight and obese adolescents and to investigate the impact of obesity on the cardiovascular system. Methods : In all, 684 adolescents were included in the study. Blood pressure, body mass index (BMI), fasting blood glucose, total cholesterol, triglyceride, low-density-lipoprotein (LDL)-cholesterol, high-density-lipoprotein (HDL)-cholesterol, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and high-sensitive C-reactive protein (hs-CRP) were measured in the patients with a BMI of >85 percentile. Brachial-ankle pulse wave velocity (BaPWV) and ankle brachial index were measured using Vascular Profiler (VP)-1000. Results : MS was confirmed in 19.5% of the overweight and obese adolescents and 50.8% of the obese adolescents. The systolic and diastolic blood pressure, height, weight, fat mass, %fat, BMI, obesity index, and waist circumference were higher in the overweight and obese adolescents with MS. Moreover, the triglyceride, AST, ALT, and hs-CRP levels were higher, whereas HDL-cholesterol level was significantly lower in this group. The overweight and obese adolescents with MS showed shorter diastolic and systolic times, higher heart rate and BaPWV, and longer E-wave deceleration time by echocardiography. Conclusion : Overweight and obese adolescents showed characteristic MS features such as hypertension and hyperlipidemia. Thus, obese adolescents predisposed to MS should be provided early treatment for obesity.

Interaction of Forskolin with the Effect of $N^6-Cyclopentyladenosine$ on $[^3H]-Acetylcholine$ Release in Rat Hippocampus (흰쥐 해마에서 Acetylcholine 유리에 미치는 $N^6-Cyclopentyladenosine$ 및 Forskolin의 영향)

  • Choi, Bong-Kyu;Park, Hie-Man;Kang, Yeon-Wook;Kook, Young-Johng
    • The Korean Journal of Pharmacology
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    • v.28 no.2
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    • pp.129-136
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    • 1992
  • As it has been reported that the depolarization-induced acetylcholine (ACh) release is modulated by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus and various lines of evidence indicate the involvement of adenylate cyclase system in $A_1-adenosine$ post-receptor mechanism in hippocampus, it was attempted to delineate the role of adenylate cyclase system in the $A_1-receptor-mediated$ control of ACh release in this study. Slices from rat hippocampus were incubated with $[^3H]-choline$ and the release of the labelled products was evoked by electrical stimulation $(3\;Hz,\;5\;Vcm^{-1},\;2\;ms,\;rectangular\;pulses)$, and the influence of various agents on the evoked tritium-outflow was investigated. $N^6-cyclopentyladenosine$ (CPA), a specific $A_1-adenosine$ receptor agonist, in concentrations ranging from 0.1 to $10\;{\mu}M$, decreased the $[^3H]-ACh$ release in a dose-dependent manner without the changes of basal rate of release. 8-cyclopentyl-1,3-dipropylxanthine $(DPCPX,\;1{\sim}10\;{\mu}M)$, a selective $A_1-receptor$ antagonist, increased the $[^3H]-ACh$ release in a dose-related fashion with slight increase of basal tritium-release. And the CPA effects were significantly inhibited by DPCPX $(2\;{\mu}M)$ pretreatment and the dose-response curve produced by CPA was shifted to the right. The responses to N-ethylmaleimide $(NEM,\;10\;&\;30\;{\mu}M)$, a SH-alkylating agent of G-protein, were characterized by increments of the evoked ACh-release and the basal release, and the CPA effect were completely abolished by NEM pretreatment. Forskolin, a specific adenylate cyclase activator, in concentrations ranging from 0.3 to $10\;{\mu}M$, increased the evoked ACh-release in a dose-dependent manner and the CPA effects were inhibited by forskolin. These results indicate that the $A_1-adenosine$ heteroreceptor plays an important role in ACh-release via nucleotide-binding protein Gi in the rat hippocampus and that the adenylate cyclase system might be participated in this process.

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Studies on the Interactions of $M_1-,M_2-receptors$ with Nicotinic Receptors in Rabbit Sympathetic Ganglia (가토 교감신경절에서 무스카린성 수용체 아형과 니코틴성 수용체의 상호작용에 대한 연구)

  • Kim, Jong-Keun;Rhu, Choon-Sik;Kang, Sam-Suk
    • The Korean Journal of Pharmacology
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    • v.28 no.2
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    • pp.171-179
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    • 1992
  • Effects of a $M_1$ receptor antagonist, pirenzepine, a $M_2$ receptor antagonist, AF-DX116, and a nicotinic receptor antagonist, mecamylamine on the pressor responses to preganglionic sympathetic nerve stimulation (PNS) and McN-A-343 and DMPP in spinal (pithed) rabbits were investigated, in order to elucidate a functional role of $M_1$, $M_2$ and nicotinic receptors in ganglionic transmission. Pirenzepine and AF-DX116 selectively inhibited the McN-A-343-induced pressor response in chlorisondamine-treated rabbit and the BCh-induced bradycardia, respectively. Electrical stimulations of preganglionic sympathetic outflow at T8 level produced increases in blood pressure. Pirenzepine $(3\;{\mu}g/kg)$ significantly inhibited the PNS-induced pressor response and the degree of inhibition was not changed by increasing the doses to $100\;{\mu}g/kg$. AF-DX116 $(100\;{\mu}g/kg)$ had no effect on the PNS-induced pressor response. Mecamylamine inhibited the PNS-induced pressor response in a dose-dependent manner. The inhibitory action of mecamylamine was significantly augmented by combined-treatment with pirenzepine $(30\;{\mu}g/kg)$ but AF-DX116 $(100\;{\mu}g/kg)$ did not affect the inhibitory action of mecamylamine. McN-A-343 and DMPP elicited pressor response in the spinal rabbit. Pirenzepine and AF-DX116 dose-dependently inhibited the McN-A-343-induced pressor response but they did not affect DMPP-induced pressor response. Mecamylamine inhibited both pressor responses induced by McN-A-343 and DMPP. These results suggest that not only nicotinic receptors but also $M_1$ receptors play a facilitatory role in ganglionic transmission but $M_2$ receptors do not contribute the transmission in spinal (pithed) rabbits.

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Analysis of Wind Vorticity and Divergence in the High-latitude Lower Thermosphere: Dependence on the Interplanetary Magnetic Field (IMF) (고위도 하부 열권 바람의 소용돌이도와 발산 분석: 행성간 자기장(IMF)에 대한 의존도)

  • Kwak, Young-Sil;Lee, Jae-Jin;Ahn, Byung-Ho;Hwang, Jung-A;Kim, Khan-Hyuk;Cho, Kyung-Seok
    • Journal of Astronomy and Space Sciences
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    • v.25 no.4
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    • pp.405-414
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    • 2008
  • To better understand the physical processes that control the high-latitude lower thermospheric dynamics, we analyze the divergence and vorticity of the high-latitude neutral wind field in the lower thermosphere during the southern summertime for different IMF conditions. For this study the National Center for Atmospheric Research Thermosphere-Ionosphere Electrodynamics General Circulation Model (NCAR-TIEG CM) is used. The analysis of the large-scale vorticity and divergence provides basic understanding flow configurations to help elucidate the momentum sources that ulti-mately determine the total wind field in the lower polar thermosphere and provides insight into the relative strengths of the different sources of momentum responsible for driving winds. The mean neutral wind pattern in the high-latitude lower thermosphere is dominated by rotational flow, imparted primarily through the ion drag force, rather than by divergent flow, imparted primarily through Joule and solar heating. The difference vorticity, obtained by subtracting values with zero IMF from those with non-zero IMF, in the high-latitude lower thermosphere is much larger than the difference divergence for all IMF conditions, indicating that a larger response of the thermospheric wind system to enhancement in the momentum input generating the rotational motion with elevated IMF than the corresponding energy input generating the divergent motion. the difference vorticity in the high-latitude lower thermosphere depends on the direction of the IMF. The difference vorticity for negative and positive $B_y$ shows positive and negative, respectively, at higher magnetic latitudes than $-70^{\circ}$. For negative $B_z$, the difference vorticities have positive in the dusk sector and negative in the dawn sector. The difference vorticities for positive $B_z$ have opposite sign. Negative IMF $B_z$ has a stronger effect on the vorticity than does positive $B_z$.

Effect of $Ca^{2+}$ and $Ca^{2+}-antagonists$ on the Spontaneous Contractions and Electrical Activities of Guinea-pig Stomach (기니피그 위 평활근의 자발적 수축과 전기적 활동에 대한 $Ca^{2+}$$Ca^{2+}$-길항제 영향)

  • Rhie, Sang-Ho;Kim, Ki-Whan
    • The Korean Journal of Physiology
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    • v.21 no.2
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    • pp.241-257
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    • 1987
  • The effects of external $Ca^{2+}$ and $Ca^{2+}-antagonists$ on the spontaneous contractions and electrical activities were investigated in guinea-pig stomach in order to clarify the mechanism for the generation of slow waves. Electrical responses of circular smooth muscle cells were recorded using glass capillary microelectrodes filled with 3 M KCl. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows: 1) The amplitude of spontaneous contractions was maximal at around 2-4 mM $Ca^{2+}$, whereas their frequency was inversely related with external $Ca^{2+}$ within the range of 0.5 to 16 mM $Ca^{2+}$. 2) Verapamil suppressed the amplitude of spontaneous contraction in a dose-dependent manner, while the frequency of spontaneous contractions was almost not changed over the whole concentration of verapamil $(0.01{\sim}5\;mg/l)$. 3) Manganese increased both the amplitude and the frequency of spontaneous contractions dose-dependently in low $Mn^{2+}$ (below 0.05 mM $Mn^{2+}$), while their amplitude and frequency were decreased in high $Mn^{2+}$ (above 0.1 mM $Mn^{2+}$). 4) The ampltude and maximum rate of rise of slow waves were incrased in high $Ca^{2+}$ solution. In $Ca^{2+}-free$ solution, the spontaneous contractions recorded simultaneously with slow waves ceased and tonic contraction ($Ca^{2+}-free$ contracture) was developed in parallel with membrane depolarization and the disappearance of slow waves. 5) Verapamil (1 mg/1) decreased the amplitude and maximum rate of rise of slow waves and it depolarized the membrane by about 6 mV, whereas the frequency of slow waves was not affected by verapamil. 6) Manganese showed different characteristic effects between low and high $Mn^{2+}$ on the slow waves: In low $Mn^{2+}$ (0.05 mM $Mn^{2+}$), the initial rapid increases and the subsequent gradual decreases in three parameters of slow waves (amplitude, rate of rise, and frequency of slow waves) till a new steady state were observed. However, in high $Mn^{2+}$ (0.5 mM $Mn^{2+}$) slow waves disappeared and membrane was depolarized. From the above results, the following conclusions could be made: 1) $Ca^{2+}$ is necessary for a generation of the slow waves, even though it is small amount. 2) Verapamil suppresses the spontaneous contractions of gastric antral strip by the decreases in amplitude and maximum rate of rise of slow waves, while this drug does not block the $Ca^{2+}-channel$ involved in the generation of slow waves. 3) Manganese has dual actions on the $Ca^{2+}-channels$; the $Ca^{2+}-channel$ involved in the generation of slow waves (or Na-Ca exchange system) or the channel for the generation of spike potentials are stimulated by a low concentration of $Mn^{2+}$, while both the $Ca^{2+}$. Channels are blocked by high concentration of $Mn^{2+}$.

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Relation of Ethanol and Calcium to Contractile and Electrical Activity of Cat Stomach (고양이 위(胃)의 수축 및 전기활동에 대한 에탄올과 칼슘의 관계)

  • Kim, Myung-Suk;Sim, Sang-Soo;Yoon, Shin-Hee;Han, Sang-Jun;Kim, Chung-Chin;Choi, Hyun
    • The Korean Journal of Physiology
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    • v.21 no.2
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    • pp.259-272
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    • 1987
  • This was study carried out to investigate the effect of calcium on spontaneous contraction and electrical activity induced by ethanol in gastric smooth muscle. After peeling off the mucous membrane from the isolated whole stomach of 102 cats, two kinds of small muscle preparations $(2.0{\times}0.2\;cm)$, one longitudinal and the other circular, were excised from the fundus, the corpus and the antrum portion of each whole stomach specimen. The isometric contraction of the small muscle preparation was measured in a cylinder-shaped chamber filled with Krebs-Ringer-dextrose solution (pH 7.4, temperature $36{\pm}0.5^{\circ}C$) bubbling with 5% $CO_2$ in $O_2$. A large muscle preparation $(5.0{\times}1.2\;cm)$ was excised from the anterior wall of the corpus-antrum portion of the same specimen in 72 of 102 cats. The gastric electrical activity (slow wave and spike potential) was monopolarly recorded by four capillary electrodes (Ag-AgCl), of which two were placed on the corpus and two on the antrum, in a muscle chamber filled with the same solution as described above. Changes in the amplitude of the contraction, frequency of the gastric slow wave and the production of the spike potential were observed after adding ethanol and/or under the treatments with verapamil, $CaCl_2$ and Ca-free Krebs-Ringer-dextrose solution. The results were as follows: 1) After adding ethanol, the spontaneous phasic contraction of the corpus was reduced dose-dependently (0.125-2.0%), which was totally abolished by higher concentrations (2.0-8.0%) of ethanol. 2) The corporal phasic contraction was also completely abolished by verapamil $(3{\times}10^{-5}\;M)$ or Ca-free Krebs-Ringer-dextrose solution. The contraction was increased by $CaCl_2\;(1.8{\times}10^{-3}\;M)$, but the inhibitory effect of ethanol on the contraction persisted even under the treatment with $CaCl_2$. 3) At higher concentrations, ethanol caused tonic contraction of both preparations from the fundus, the corpus and the antrum in a dose-dependent manner. The tonic contraction of the fundus produced by ethanol was not influenced by $CaCl_2$ or verapamil, whereas the tonic contraction was not produced by ethanol in tile Ca-free solution. 4) Frequency of gastric slow wave was decreased dose-dependently by the addition of ethanol (0.25-1.0%), and tile slow wave was not produced by higher concentration of ethanol (2.0%). 5) The frequency of slow wave was significantly reduced by verapamil only and the inhibitory influence of ethanol on the slow wave frequency was reinforced by verapamil. 6) The treatment of $CaCl_2$ increased significantly the slow wave frequency, and attenuated the inhibitory effect of ethanol on the frequency. It is therefore suggested that ethanol regulates the phasic contraction and the production of slow wave by interfering with the transport of calcium in the stomach muscle of the cat.

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A Study on the Current Fire Insurance Subscription and Solutions for Ensuring the Safety of the Traditional Market (전통시장 안전성 확보를 위한 개선방안: 화재보험 가입실태를 중심으로)

  • Kim, Yoo-Oh;Byun, Chung-Gyu;Ryu, Tae-Chang
    • Journal of Distribution Science
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    • v.9 no.4
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    • pp.43-50
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    • 2011
  • Concerning the risk factors of the outbreak of a fire in a traditional market, most of those markets are located in downtown areas or residential areas; thus, although their location may be favorable in terms of marketability, they face a potential risk in that a fire may develop into a large blaze owing to poor environment or the absence of facilities prepared for disaster during a fire. Moreover, as many people are densely poised in the markets, it is very probable that a fire may occur owing to the excessive use of heaters in the winter as well as the reckless use of electric and gas facilities. It seems that traditional markets encounter difficulty being insured against fire, because of their vulnerability and that the vast majority of small-scale sellers are likely to suffer mental anguish and tremendous physical injury in case of a fire. However, most of those sellers in the traditional markets are hand-to-mouth sellers, and they lack awareness of safety concerns and have insufficient experience in safe facility management. As small-scale sellers constitute the majority in the traditional market, the subscription rate of fire insurance in most of the traditional markets is low for the reasons of their needy circumstances and their financial burden. Statistically, the subscription by street vendors is non-existent; therefore, these vendors have a fairly limited access to indemnification after fire damage. Because of these problems, this study's purpose is to identify the current level of insurance subscription by these markets, which are exposed to poor facilities and vulnerability to fire. In order to fix this, it appears that shop owners and consumers will have to band together. For this study, we executed a fire policyholder fact-finding mission at traditional markets with approximately 108 and 981 stores. The research method was executed by an investigation using one-on-one individual interviews using a questionnaire. The contents investigated current insurance subscriptions. The method of analysis looked at the difference of insured amount according to volume size through cross-tabulation of the difference of insured amount by possession form, difference of insured amount by market form, difference of insured amount by category of business, difference of insured amount by market size, etc. Furthermore, the study should be used to propose solutions for problems through theoretical review with the use of a literature research, because the field case study was through interviews with the persons concerned, and the survey of the current insurance subscriptions by traditional market shopkeepers. The traditional market would generally have difficulty affording fire insurance. Fire insurance subscription rates of most of the market proved to be inactive, because of the economic burden of payment. Lack of funds is thought to be the main factor that causes a lack of realization about the necessity of fire insurance. In addition to expensive insurance premiums, sometimes, the companies' valuation of the businesses is lower than their actual valuations, and they do not pay out enough during a claim. The research presents an improvement plan that, when presented at the traditional markets, may strengthen their ability to procure fire insurance through the help of the central government. Researchers connected with the traditional market mainly accomplish the initial research. However, although this research has its limitations, it offers considerable benefits. For future researchers, I would suggest looking at several regions for comparison.

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