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A Study on the Mechanism of Insulin Sensitivity to Glucose Transport System: Distribution of Subcellular Fractions and Cytochalasin B Binding Proteins (인슐린의 포도당 이동 촉진 기전에 관한 연구 -세포내부 미세구조와 Cytochalasin B 결합단백질의 분포-)

  • Hah, Jong-Sik
    • The Korean Journal of Physiology
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    • v.24 no.2
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    • pp.331-344
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    • 1990
  • What makes glucose transport function sensitive to insulin in one cell type such as adipocyte, and insensitive in another such as liver cells is unresolved question at this time. Recently it is known that insulin stimulates glucose transport in adipocytes largely by redistributing transporter from the storage pool that is included in a low density microsomal fraction to plasma membrane. Therefore, insulin sensitivity may depend upon the relative distribution of gluscose transporters between the plasma membrane and in an intracellular storage compartment. In hepatocytes, the subcellular distribution of glucose transporter is less well documented. It is thus possible that the apparent insensitivity of the hepatocyte system could be either due to lack of the constitutively maintained, intracellular storage pool of glucose transporter or lack of insulin-mediated transporter translocation mechanism in this cell. In this study, I examined if any intracellular glucose transporter pool exists in hepatocytes and this pool is affected by insulin. The results obtained summarized as followings: 1) Distribution of subcellular fractions of hepatocyte showed that there are $24.9{\pm}1.3%$ of plasma membrane, $36.9{\pm}1.7%$ of nucleus-mitochondria enriched fraction, $23.5{\pm}1.2%$ of lysosomal fraction, $9.6{\pm}1.0%$ of high density microsomal fraction and $4.9{\pm}0.5%$ of low density microsomal fraction. 2) In adipocyte, there were $29.9{\pm}2.6%$ of plasma membrane, $19.4{\pm}1.9%$ of nucleus-mitochondria enriched fraction, $26.7{\pm}1.8%$ of high density microsomal fraction and $23.9{\pm}2.1%$ of low density microsomal fraction. 3) Surface labelling of sodium borohydride revealed that plasma membrane contaminated to lysosomal fraction by $26.8{\pm}2.8%$, high density microsomal fraction by $8.3{\pm}1.3%$ and low density microsomal fraction by $1.7{\pm}0.4%$ respectively. 4) Cytochalasin B bound to all of subcellular fractions with a Kd of $1.0{\times}10^{-6}M$. 5) Photolabelling of cytochalasin B to subcellular fractions occurred on 45 K dalton protein band, a putative glucose transporter and D-glucose inhibited the photolabelling. 6) Insulin didn't affect on the distribution of subcellular fractions and translocation of intracellular glucose transporters of hepatocytes. 7) HEGT reconstituted into hepatocytes was largely associated with plasma membrane and very little was found in low density microsomal fraction which equals to the native glucose transporter distribution. Insulin didn't affect on the distribution of exogeneous glucose transporter in hepatocytes. From the above results it is concluded that insulin insensitivity of hepatocyte may due to lack of intracellular storage pool of glucose transporter and thus intracellular storage pool of glucose transporter is an essential feature of the insulin action.

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Effect of thyroparathyroidectomy on urinary excretion of sodium, potassium, calcium and phosphate in the rabbits (갑상선 부갑상선 적출이 뇨중 Na, K, Ca 및 $PO_4$ 배설에 미치는 영향)

  • Choi, Duck-Kyung
    • The Korean Journal of Physiology
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    • v.16 no.1
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    • pp.69-76
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    • 1982
  • The effect of parathyroid hormone on calcium and phosphate metabolism have been widely investigated, however less attention has been paid to the effect on urinary excretion. This study was performed for the purpose determining urinary excretion of Na, K, Ca, and $Po_4$, of 18 thyroparathyroidectomized (TPTX) rabbits, which were TPTX previously 7 to 10 days compared with the same normal ones. After TPTX 0.2 mg/day of synthyroid was donated to the rabbits. The concentration of electrolytes in the serum and urine was determined by the following method; Na and K were determined by means of flame photometry, Ca was by EDTA titration $method^{19)}$, and $Po_4$ by Fiske and Subba-Raw $method^{20)}$. The results as follows. The concentrations of electrolytes in the serum were 1) In the normal control rabbits (N = 25) (data, $Mean{\pm}S.E.$) $Na\;131.72{\pm}1.33\;mEq/L$, $K\;3.59{\pm}0.28\;mEq/L$, $Ca\;12.58{\pm}0.29\;mg%$, $Po_4\;4.50{\pm}\;0.45mg%$. 2) In the TPTX rabbits(N= 18) $Na\;140.6l{\pm}2.56\;mEq/L$, $K\;3.38{\pm}0.36\;mEq/L$, $Ca\;l2.18{\pm}0.45\;mg%$, $Po_4\;3.92{\pm}\;0.35\;mg%$. There was no significant change between the normal and TPTX rabbits. The concentration of elelctrolytes in the urine were variously changed. 3) In the normal rabbits. $Na\;8.40{\pm}1.09\;mEq/L$, $K\;81.59{\pm}10.19\;mEq/L$, $Ca\;16.02{\pm}3.12\;mg%$, $Po_4\;13.16{\pm}2.89mg%$. 4) In the TPTX rabbits, $Na\;14.57{\pm}3.39\;mEq/L$ slight ncreased, $K\;116.06{\pm}12.77\;mEq/L$ significant increased (P<0.05), $Ca\;18.90{\pm}5.44\;mg%$ no significant increased, $Po_4\;43.38{\pm}8.67\;mg%$ significant increased (p<0.01). The effect of TPTX was assumed that it affected upon increasing tubular secretion of $K^+$ and inhibition of the tubular reabsorption of $Po_4$.

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The Analgesic Effects of Apitoxin and its Mechanism via JOR and Measuring Expression of mRNA in Phospholipase and TPH using RT-PCR (Jaw Opening Reflex 및 RT-PCR을 이용한 봉독의 진통효과)

  • Cho, Kwang-Ho;Lee, Jae-Dong;Park, Dong-Suk;Ahn, Byoung-Choul
    • Journal of Pharmacopuncture
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    • v.3 no.1
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    • pp.35-51
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    • 2000
  • The purpose of this study is to prove the analgesic effects of apitoxin and its mechanism via jaw-opening reflex(JOR) and measuring expression of mRNA in Phospholipase and Tryptophan hydroxylase(TPH) using RT-PCR. The experiments were carried out on Sprague-Dawley rats(300-400g) and mastocytoma(P-185 HTR) for JOR and RT-PCR, respectively. Rats anesthetized with thiopental sodium (80mg/kg) were used in the Tooth Pulp stimulation induced JOR. The amplitude of a digastric electromyogram (dEMG) was recorded during the stimulation at an intensity of 1.5 times the threshold for JOR. Apitoxin used in this experiment was diluted with normal saline by 1:1000. Apitoxin was injected intravenously into the test group while normal saline to the control group. However, it was injected directly into the cell of mastocytoma. We referred to base sequence registered in Genbank in designing primers for RT-PCR. The results were as follows; (1)Compared with control group, analgesic effect started to show right after Sprague-Dawely rats were treated with apitoxin($71.50{\pm}8.08$) and lasted for 50 minutes. (2)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of phospholipase or rate-limiting enzyme of biosynthesis of prostaglandins with $10{\mu}g/ml$ apitoxin.($31.74{\pm}18.98%$, P<0.05) (3)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of TPH or rate-limiting enzyme in biosynthesis of serotonin with $10{\mu}g/ml$ apitoxin.($131.37{\pm}16.87%$, P<0.05). These results suggest that $10{\mu}g/ml$ apitoxin have the most analgesic effects. This study showed that apitoxin has analgesic effects and held good for 50 minutes. The injection of apitoxin has brought out changes in the degree of expression of phospholipase and TPH. These results strongly suggest that analgesic mechanism by apitoxin is closely related to prostaglandins and serotonin.

Effect of Brine Treatment Applied in the Manufacture of Traditional Forged High Tin Bronzes of Korea (한국의 방짜유기에 가해지는 염수처리의 효과에 관한 연구)

  • Lee, Jae-Sung;Jeon, Ik-Hwan;Kwak, Seok-Chul;Park, Jang-Sik
    • Journal of Conservation Science
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    • v.28 no.4
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    • pp.403-410
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    • 2012
  • The brine treatment applied during the fabrication of forged high tin bronze objects is considered effective at the removal of surface oxide layers developed at elevated temperatures. There is not much information, however, available for the understanding of its exact effect and purpose. This work performed laboratory experiments to characterize the effect brine treatments produce on the surface of bronze objects during fabrication. Specimens were first made in the bronze shop of the Yongin folk village under varying conditions of brine treatments, and the results obtained were then used in the following laboratory experiments where the effect of brine treatments were investigated in terms of brine concentrations, alloy compositions and thermo-mechanical treatments. The results show that oxide layers generated at high temperature are easily removed by the brine treatment. It was found that the element, chlorine, played a key role in the removal of such oxide layers as opposed to the other constituent of the brine, sodium, makes no notable contribution. In bronze alloys containing 22% tin, this brine effect is obtained regardless of the application of forging as long as the brine concentration is over 0.5% based on weight. In alloys containing lead, however, no brine effect is observed due to the molten lead that emerges from inside the hot bronze specimen and forms a thin layer on its surface.

Fine Structure on the Pigment Epithelial Cell and the Bruch's Membrane of the Rat Retina after X-Irradiation (X-선 조사를 받은 흰쥐 망막의 색소상피세포와 기저복합층의 미세구조)

  • Ko, Jeong-Sik;Shin, Ki-Ho;Ahn, E.Tay;Yang, Nam-Gil;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.23 no.2
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    • pp.11-26
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    • 1993
  • This experiment was performed to study the morphological responses of the pigment epithelial cell and the Bruch's membrane of the retina of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The heads of the rats, under sodium thiopental anesthesia, were exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80cm, and the. dose rate was 200 rads/min. The experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Under anesthesia, 1% glutaraldehyde-1% paraformaldehyde solution(0.1M Millonig's phosphate buffer, pH 7.3) was perfused through the left ventricle and ascending aorta. Pieces of the tissue taken from the posterior region of the retina were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.3) and 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH7.3), and embedded in araldite mixture. The ultrathin sections contrasted with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. The morphological changes of the pigment epithelial cells were not pronounced after exposure to 3,000 rads of X-ray. But on the 6th hour after exposure to 6,000 rads of X-ray, bulging nuclear membrane protruding into the cytoplasm and nuclear chromatin clumped into numerous masses along the nuclear membrane were observed. At the 2nd and 6th day post-irradiation, partial cytolysis or necrosis were seen. 2. The thickness of the Bruch's membrane of the experimental groups were increased in the time and dose range covered by this study, and splitting or diffusing basal laminae of the choriocapillary layer were observed frequently in the experimental group. Above results suggest that large amount(6,000 rads) of head irradiation induce direct hazardous effects on the pigment epitherial cells and Bruch's membrane of the retina of the rat, but pigment epithelial cells are more radioresistant than Bruch's membrane.

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The Effect of Drinking Water Fluoride on the Fine Structure of the Ameloblast in the Fetal Rat (음용수 불소가 흰쥐태아 법랑모세포의 미세구조에 미치는 영향)

  • Lim, Do-Seon;Jeong, Moon-Jin;Yoe, Sung-Moon
    • Applied Microscopy
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    • v.29 no.2
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    • pp.189-193
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    • 1999
  • The response of ameloblast to long term (3 weeks) exposure to fluoride was examined in continuously erupting mandibular incisors of pregnancy rats as compared to control rats receiving a similar diet (Teklad L-356) but no sodium fluoride in there drinking water. Rats were started on water containing 0 ppm, 100 ppm, 200 ppm, and 300 ppm NaF at the beginning of pregnancy. To examine on the ultrastructural changes of the ameloblast, electron microscopy was used. The results indicated that rat incisors expressed two major changes in normal amelogenesis that could be attributed to chronic fluoride treatment. The fluoride produces marked alteration in the fine structure of ameloblast from teeth of young rats, such as large confluent distensions of the endoplasmic reticulum and swelling of isolated mitochondria, in particular on the morphology of the rough-surfaced endoplasmic reticulum. A graded series of alterations to these organelles were produced, and the severity of the changes would seem to be dependent on dose and time. This experimental data suggested that exposure prolonged of animal to high level of fluoride appears to induce morphological changes in the normal appositional growth and initial mineralization of enamel created during amelogenesis.

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Ultrastructural Study on the Ependymal Cells of the Read-Irradiated Rats (방사선이 뇌실막세포의 미세구조에 미치는 영향)

  • Ahn, E-Tay;Cho, Hwee-Dong;Kim, Jin-Gook;Park, Kyung-Ho;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.29 no.1
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    • pp.11-23
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    • 1999
  • Ultrastructure of the ependymal cells of X-irradiated rats on their head were studied. Rats weighing $200\sim250gm$ were X-irradiated on their head and neck areas. Total exposures were 3,000 rads or 6,000 rads depending on experimental groups. And irradiated rats were sacrificed on 6 hours, 2 days and 6 days following the radiation exposures. Animals were perfused through the heart with 1% glutaraldehyde-1% paraformaldehyde solution, under ether-anesthesia. The tissues from the wall of lateral ventricles were fixed in the 2% osmium tetroxide solution. The results observed with electron microscope were as follow: 1. In 6 hours group, many ependymal cells were swelled, luminal portions of cytoplasms of some cells protruded into the ventricular lumen, and many cilia were lost or irregularly altered. 2. In 2 days group, ependymal cells were swelled more severely and subependymal edema were pronounced. 3. Protruded cytoplasm contained usually basal bodies of cilia, groups of mitochondria, endoplasmic reticula , etc. 4. Following X-irradiations, some protruded masses contained neural elements including the axon terminals with dense core vesicles. Axons and axon terminals were also found in the enlarged intercellular spaces among ependymal cells. From the above results, the heavy irradiation on the head area of the rat induced alteration of the ependymal cells lining the lateral ventricle. Hence the ependymal functions of selective barrier, protective barrier, and metabolic barrier could be altered following X-irradiation on the head.

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The Daily Intakes of Nutrients and Snacks of High School Smokers and Non-Smokers (고등학생 흡연자와 비흡연자의 1일 영양소 및 간식 섭취)

  • Song, Young-Mi;Han, Jang-Il;Kim, Seong-Ai
    • Korean Journal of Community Nutrition
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    • v.13 no.4
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    • pp.476-488
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    • 2008
  • This study investigates the relationship of smoking on daily intake of nutrients and snacks in the Chungnam and Daejeon high school students. Up to date scientific nutrition education and counseling programs in the regular school system is needed for a professional nutrition education teacher. The primary objective of this study was to provide useful information to nutrition education teachers. A survey was conducted with 400 high school students in the Chungnam and Daejeon areas. 381 out of 400 questionnaires were analyzed with SPSS 12.0K. The subjects were composed of 49.8% male, 50.1% female and 40.9% regular high school students, 59.1% business high school students and smokers 43.1%, non-smokers 56.9%. 43.4% of smokers had been smoking since middle school. On analysis of daily nutrient intakes, 16 out of 19 nutrients except animal calcium, Vitamin A and Vitamin C were much more consumed by the smoking group than the non-smoking group non-significantly. Especially vegetable fat and Vitamin E were higher in the smoking group than the non-smoking group (p<0.05). The intake ratio of carbohydrates: protein: fat was similar in the two groups (smoking group 55:15:29, nonsmoking group 56:15:28). Intakes of Vitamin B1 and potassium in comparison with the Korean dietary reference intakes(KDRI) were under 50% in both groups. However, sodium was taken over 200% compared to KDRI in both groups. Intakes of Vitamin C in the smoking group were as low as 76.5% in comparison to KDRI. Smokers need to increase the intakes of Vitamin C considering that smokers need to intake Vitamin C two times than non-smokers. Nutrient intakes from snacks in the smoking group were higher than the non-smoking group. Nutrients that originated from snacks which took over 20% among daily nutrient intakes were 12 nutrients(energy, fat, carbohydrate, calcium, P, Fe, K, Vitamin $B_1$, Vitamin $B_2$, Vitamin C, Vitamin E, dietary fiber) in the smoking group compared to 7 nutrients(energy, vegetable protein, fat, carbohydrate, calcium, Vitamin $B_2$, Vitamin C) in the non-smoking group. The smoking group was significantly paying more money for snacks each month than the non-smoking group was(p<0.01). Periods of consumption were irregular in the smoking group(p<0.05) and the smoking group was used to taking snacks in the morning compared to the non-smoking group. The smoking group preferred sweets and high calorie food over other snacks in comparison of the non-smoking group. The nonsmoking group had better eating habits than the smoking group.

Dietary and Non-dietary Factors Related to Bone Mineral Density in Female College Students (여대생의 골밀도와 식이 및 비식이 요인들 간의 상관성)

  • Lim, Ji-Hye;Bae, Hyun-Sook;Lee, Seung-Min;Ahn, Hong-Seok
    • Korean Journal of Community Nutrition
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    • v.13 no.3
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    • pp.418-425
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    • 2008
  • The purpose of this study was to investigate correlations between bone mineral density (BMD) and dietary and non-dietary factors in female college students. The BMD of the subjects (n = 38) was measured using DEXA (Dual Energy X-ray Absorptiometry) at lumbar spine and three femoral sites including femoral neck, ward's triangle, and femoral trochanter. Three-day 24-hour dietary recall data were collected from each subject to assess consumption levels of nutrients and food groups. The mean (${\pm}SD$) values of age, age of menarche, height, weight, body mass index (BMI), waist-to-hip ratio (WHR), fat mass, and % body fat of the subjects were 21.34 (${\pm}1.73$) years, 13.1 (${\pm}1.2$) years, 161.3 (${\pm}5.0$) cm, 53.7 (${\pm}7.2$) kg, 20.6 (${\pm}2.6$) $kg/m^2$, 0.80 (${\pm}0.04$), 15.4 (${\pm}4.4$) kg, and 28.2 (${\pm}4.7$), respectively. The BMD values of lumbar spine, femoral neck, ward's triangle, and femoral trochanter as T-value were $1.150{\pm}0.13\;g/cm^2$, $0.932{\pm}0.11\;g/cm^2$, $0.850{\pm}0.13\;g/cm^2$, and $0.721{\pm}0.10\;g/cm^2$, respectively. The daily mean energy intake of the subjects was 1660.6 kcal. The intake levels of carbohydrate, calcium, iron, vitamin C, and folic acid were lower than the KDRIs, while those of fat, phosphorus, sodium, vitamin A, and vitamin $B_6$ were higher than the KDRIs. Significantly negative correlation were detected between consumption of fat and oils and the BMD of all sites measured (p < 0.05). Potato and starch intake was negatively correlated to the BMD of femoral neck and word's triangle (p < 0.05). The intake of cereals was found to be negatively correlated to the BMD of ward's triangle (p v 0.05). There was also negative correlation of intake of soybeans and products with the BMD of lumbar spine (p < 0.05). Weight and muscle mass were positively correlated to the BMD of both lumbar spine and femurs (p < 0.01). Significantly positive correlations between BMI, and fat mass and the BMD of lumbar spine were observed (p < 0.001). Additionally fat mass was positively correlated to the BMD of femoral neck. WHR and % body fat were positively correlated to the BMD of lumbar spine (p < 0.05). Negative correlation was found between a serum calcium level and the BMD of femoral neck (p < 0.05). In summary non-dietary factors generally showed significant correlations with the BMD of lumbar spine, while several dietary factors showed significant correlations with the BMD of femurs.

Associations between Exposure to Unhealthy Food Outlets Within Residential District and Obesity: Using Data from 2013 Census on Establishments and 2013-2014 Korea National Health and Nutrition Examination Survey (거주지 주변의 식품환경과 비만의 관련성 연구: 2013 전국사업체조사와 2013-2014 국민건강영양조사 자료를 이용하여)

  • Kim, Yoonjung;Han, Sung Nim
    • Korean Journal of Community Nutrition
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    • v.21 no.5
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    • pp.463-476
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    • 2016
  • Objectives: Environmental, social and personal factors influence eating patterns. This study aimed to investigate the relationship between unhealthy food outlets within a residential area and obesity using nationally representative Korean survey data and data from the Census on Establishments. Methods: Data on the food intakes and socioeconomic variables of a total of 9,978 adults aged ${\geq}19$ years were obtained from the 2013-2014 Korea National Health and Nutrition Examination Survey. Geographic locations of restaurants were obtained from the 2013 Census on Establishments in Korea. Administrative area was categorized into tertiles of count of unhealthy food outlets based on the distribution of number of unhealthy food outlets among all urban (Dong) and rural (Eup or Myun) administrative districts in Korea. Multilevel logistic regressions model were used to assess the association between the number of unhealthy food outlets and obesity. Results: People living in the district with the highest count of unhealthy food outlets had higher intakes of fat (45.8 vs. 44.4 g/day), sodium (4,142.6 vs. 3,949.8 mg/day), and vitamin A (753.7 vs. $631.6 {\mu}gRE/day$) compared to those living in the district with the lowest count of unhealthy food outlets. A higher count of unhealthy food outlets was positively associated with frequent consumption of instant noodles, pizza, hamburgers and sandwiches, sweets and sour pork or pork cutlets, fried chicken, snacks, and cookies. Higher exposure to unhealthy food outlets was associated with increased odds of obesity (1st vs. 3rd tertile; OR 1.689; 95% CI 1.098-2.599). Conclusions: A high count of unhealthy food outlets within a residential area is positively associated with the prevalence of obesity in Korea. The results suggest that food environmental factors affects the health outcomes and interventions aiming to restrict the availability of unhealthy food outlets in local neighborhoods may be a useful obesity prevention strategy.