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Cadms/SynCAMs/Necls/TSLCs Interact with Multi-PDZ Domain Protein MUPP1 (Cadms/SynCAMs/Necls/TSLCs와 multi-PDZ domain protein MUPP1 단백질의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1276-1283
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    • 2014
  • Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.

Characterization of a Mucolipidosis Type II Mouse Model and Therapeutic Implication of Lysosomal Enzyme Enriched Fraction Derived from Placenta (뮤코지방증 2형 마우스 모델의 특징과 태반에서 추출한 리소좀 효소 투여의 결과)

  • Cho, Sung Yoon;Kim, Ki-Yong;Kim, Su Jin;Sohn, Young Bae;Maeng, Se Hyun;Kim, Chi Hwa;Ko, Ah-Ra;Song, Junghan;Yeau, Sung-Hee;Kim, Kyung-Hyo;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.1
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    • pp.5-13
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    • 2012
  • I-cell disease (mucolipidosis type II; MIM 252500) and pseudo-Hurler polydystrophy (mucolipidosis type III; MIM 252600) are disorders caused by abnormal lysosomal transport in cells. The presence of numerous inclusion bodies in the cytoplasm of fibroblasts, a lack of mucopolysacchariduria, increased lysosomal enzyme activity in serum, and decreased GlcNAc-phosphotransferase activity are hallmark. Here, we attempted to investigate phenotypical and biochemical characteristics of the knockoutmouse of GlcNAc-phosphotransferase ${\alpha}/{\beta}$ subunits; in addition, we also attempted to determine whether the lysosome enriched fraction derived from placenta can be beneficial to phenotype and biochemistry of the knockout mouse.We found that the knockout mouse failed to thrive and had low bone density, as is the case in human. In addition, skin fibroblasts from the animal had the same biochemical characteristics, including increased lysosomal enzyme activity in the culture media, in contrast to the relatively low enzyme activity within the cells. Intravenous injection of the lysosome rich fraction derived from placenta into the tail vein of the animal resulted in a gain of weight, while saline injected animals didn't.In conclusion, our study demonstrated the phenotypical and biochemical similarities of the knockout mouse to a mucolipidosis type II patient and showed the therapeutic potential of the lysosome enriched fraction. We admit that a larger scale animal study will be needed; however, the disease model and the therapeutic potential of the lysosome enriched fraction will highlight the hope for a novel treatment approach to mucopolipidosis type II, for which no therapeutic modality is available.

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$^{99m}Tc$-Glucarate Uptake in Ischemic Tissue of Experimental Models of Cerebral Ischemia (실험적 뇌허혈증 모델에서 허혈 조직의 $^{99m}Tc$-glucarate 섭취)

  • Jeong, Jae-Min;Kim, Young-Ju;Choi, Seok-Rye;Kim, Chae-Kyun;Mar, Woong-Chun;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon;Lee, Dong-Soo
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.4
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    • pp.484-492
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    • 1996
  • To detect ischemic tissue in experimental model of cerebral ischemia made by middle cerebral artery(MCA)-occlusion, we acquired triple image of $^{99m}Tc$-glucarate, [$^{18}F$]fluoro-deoxyglucose (FDG), and 2,3,5- triphenyltetrazolium (TTC) staining. We made cerebral infarction either with reperfusion (after occlusion of 2 hours) or without reperfusion in 10 Sprague-Dawley rats by inserting thread to MCA through internal carotid artery. After 22 hours, we injected 740 MBq of $^{99m}Tc$-glucarate and 55.5 MBq of [$^{18}F$]FDG through tail vein. Each 1 mm slice of rat brains was frozen and exposed to imaging plate for 20 minutes in freezer to get an [$^{18}F$]FDG image. After 20 hours enough to fade radioactivity of [$^{18}F$]FDG, the slices were again imaged by BAS1500 for $^{99m}Tc$-glucarate uptake. Finally, these brain tissues were stained with TTC. Semi-quantitative visual analysis was done by grading 0 to 3 points according to the degree of uptakes($^{99m}Tc$-glucarate) and decreased uptakes([$^{18}F$]FDG and TTC). Ten rats survived with neurologic symptoms. TTC staining confirmed the development of infarction. The size of the infarction was relatively larger in the group without reperfusion. [$^{18}F$]FDG images were similar to TTC-stained images. However, we found regions with intermediate uptake which were not stained with TTC. We found regions with intermediate [$^{18}F$]FDG uptake where TTC staining was normal. $^{99m}Tc$-glucarate uptake was round only in TTC non-stained region. In the TTC stained regions, there were no uptake of $^{99m}Tc$-glucarate. We could not find clear relation between $^{99m}Tc$-glucarate uptake with [$^{18}F$]FDG uptake. This was partly because percent uptake of $^{99m}Tc$-glucarate was so small (less than 1 percent of injected dose) and because there were quite heterogeneity of patterns of [$^{18}F$]FDG uptake and TTC. With these findings, we could conclude that $^{99m}Tc$-glucarate were taken up only in part of ischemic tissues which were proven to be nonviable. The establishment of MCA-occluded rat model with or without reperfusion and triple imaging for $^{99m}Tc,\;^{18}F$ and TTC helped the characterization of $^{99m}Tc$-glucarate uptakes. Further work is needed to clarify the meaning or diversities or [$^{18}F$]FDG and TTC and their relation with $^{99m}Tc$-glucarate.

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Construction of a Fluorescently Labeled Infectious R Peptide-Less Moloney MLV Molecular Clone for Analysis of Syncytium (합포체 분석을 위해 R 펩타이드가 결여된 형광 표지 Moloney 마우스레트로바이러스 Molecular Clone 제조)

  • Lee, Yong-Jin;Park, Jin-Woo;Lee, Kyu-Jun;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.246-250
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    • 2009
  • Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.

The survival rate of fish discarded from trawler (트롤 어선에서 어획후 투기된 어류의 생존율)

  • 안희춘;양용수;박창두;조삼광;박해훈;정의철
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.39 no.2
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    • pp.87-91
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    • 2003
  • Experimental fishing was carried out to investigate the survival rate of fishes discarded after hauling from the trawl and to develop the fishing gear and method for fisheries management which can improve the survival rate of young fishes escaped from the trawl codend in offshore korean southern sea and off Cheju Island of Korea. The young fishes were bred in fish cage on the board to measure the sustainable survival time for fishes escaped from grid panel and codend. The obtained results are summarized as follows' 1. Japanese flying squid(Todarodes pacjficus), mitra squid(Loligo chinesis), hair-tail( Trichiurus lepturus), john dory(Zeus japonicus), spanish mackerel(Scomberomorus niphonius), redlip croaker (Pseudosciaena polyactis) and blackthroat seaperch(Doederliinia berycoides) were dead instantly after hauling. 2. Survival rate of tiger shark(Galeocerdo cuvier), conger eel (Conger myriaster), red skate (Dasyatis akajei), black scraper(Navodon modestus) and japanese fan lobster(Ibacus ciliatus) might be high after discarding because they survived for long hours in fish cage. 3. Blotched eelpout(Zoarces gili) escaped from the escapement device(grid) was dead within 6 hours in the water tank installed on the board but 97% of tiger shark and 72% of conger eel survived over 72 hours. 4. Red skate escaped from trawl codend was dead within 60 hours in the water tank installed on the board but sustainable survival time of 25% of octopus(Paroctopus dofleini) and 100% of black scraper was over 72 hours. 5. Compared with the survival rate of rosefish(Helicolenus higendorfi) escaped from the escapement device(grid) and trawl codend, all of the rosefish escaped from the grid was dead within 7.2 hours but 8.3% of the rosefish escaped from the codend survived over 72 hours. 6. In comparison with the survival rate of japanese fan lobster survived over 72 hours, survival rate of japanese fan lobster escaped the codend was much higher as 75% than 33% of japanese fan lobster escaped from the grid.

A Demonstrative Study on the Intake Habits of Dairy Goats (Saanen) Fed with Roughages (유산양의 조사료 채식습성에 관한 실증적 연구)

  • Gang, Byung-Ho;Lee, In-Duk;Lee, Hyung-Suk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.1
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    • pp.63-72
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    • 2009
  • The experiment was conducted from 2007 to 2008. The experimental roughages include five sources and 25 species in all; grasses and legumes: 5 species (mixed grasses, orchardgrass, tall fescue, alfalfa, white clover), native grasses and weeds: 5 species (mixed native grasses, Miscanthus sinensis Anderss, Arundinella hirta (Thunb.) Tanaka, barnyard grass, short awn, forage crops and straw: 5 species (barley 4- hairy vetch, wheat 4- hairy vetch, rye silage, barley silage, baled rice straw), browse and fallen leaves: 5 species (mixed browse, oriental white oak browse, Quercus serrta Thunb., browse oriental cherry fallen leaves, Japanese chestnut fallen leaves), and imported hay and straw: 5 species (timothy hay, tail fescue straw, annual ryegrass straw, klinegrass hay, alfalfa hay). Ten dairy goats (Saanen) were selected which had nearly the same body weight (25kg). The experiment was carried out on the dairy goats farm at Geumsan-Cun in Chungnam province. The chemical composition and dry matter digestibility of each roughage source and species were significantly different at the sampling area, plant species, growth stages and cutting period. Among all the 25 species of roughages, the favorite intake species order by dairy goats was observed like this: mixed grasses, white clover, alfalfa and the lower intake species order was baled rice straw and rye silages. The dairy goats ate more roughages which had low fibrous contents, but high dry matter digestibility. On the other hand, compared to each roughage source, the goat's favorite roughage were grasses and legumes (34.6%) among the trial species. Based on the result, it is reconfirmed that the food habit of dairy goats seems to be closer to that of graters.

A Survey on the Actual Condition for Dining-out in Pusan 2. The Preference of Restaurant and Food According to Age Groups and Sex Distinction (부산지역의 외식실태조사 2. 연령과 성별에 따른 음식점과 음식의 선호도)

  • 김두진;임효진
    • The Korean Journal of Food And Nutrition
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    • v.11 no.2
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    • pp.200-210
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    • 1998
  • This study was conducted to find out the actual condition of dining-out and the preferences of restaurant and food according to age groups and sex distinction in Pusan area. The survey was conducted on 564 peoples who live or work, including students aged more than 4th grade in elementary school in Pusan area. The results of the questionnaires are as follows ; The reasons of prefferring restaurant were in order of "delicious taste", "comfortable atmosphere" and "economical price", and of no prefferring restaurant were in order on "untasty", "unsanitary" and "unkind employees". Female and student group were more in the items of unexperinced food than male and older group. The menu of preferring were in order of "sliced raw fish", "pizza", "bulgogi", and of no preferring were in order of "dog soup", "gopchang jengol", "ox tail soup" male and older group showed a relatively highest preference for "traditional food", however, female and younger(student group and young group) showed a relatively highest preference for "fast foods". The reasons of prefferring food were in order of "delicious taste", "something like" and "abundance of nutrition", and of no prefferring food were in order of "untasty", "hatred food" and "doesn't suit constitution". Male and older group showed a relatively highest preference for "abundance of nutrition", however, female and younger showed a relatively highest preference for "foreign food". But, the results of the preferences of restaurant and food were different according to sex distinction and age groups.ot;, however, female and younger showed a relatively highest preference for "foreign food". But, the results of the preferences of restaurant and food were different according to sex distinction and age groups.ge groups.

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Effects of $H_2O$-fraction of Dioscorea japonica Thunb and selenium on lipid peroxidation in streptozotocin-induced diabetic rats (참마 $H_2O$ 분획물과 Selenium 보충이 당뇨 흰쥐의 지질과산화에 미치는 영향)

  • 김명화
    • Korean journal of food and cookery science
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    • v.17 no.4
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    • pp.344-352
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    • 2001
  • The purpose or this study was to investigate the effect or H $_2$O fraction or Dioscorea japonica Thunb(DJT) and selenium(Se) on the lipid peroxidation in streptozotocin (STZ)-induced diabetic rats. Male Sprague-Dawley rats weighing 180∼220g were divided into 5 groups: One normal rat group and 4 diabetic rat groups(the STZ-Control group, the DJT group, the DJT-Se group and the Se group). Diabetes was induced in the male rats by injection of STZ into tail vein at a dose of 45 mg/kg. The H$_2$O-fraction of DJT(500 mg/kg) was administered orally for 14 days. The supplementation was achieved with the AIN-76 recommendation diet by adding 2 mg/kg diet of selenium as Na$_2$SeO$_3$ which was prepared freshly everyday. The levels of glycogen in liver and muscle and protein in kidney, liver and muscle were measured. The liver concentrations of cholesterol and triglyceride were analyzed. Also, the malondialdehyde(MDA) levels in kidney, liver and lung were determined. The glycogen levels of liver and muscle in diabetic groups were not significantly different from the normal group. The protein concentrations of kidney, liver and muscle were not significantly different either, but the level of muscle protein was higher than STZ-Control group. The levels of liver cholesterol were significantly different between normal and STZ-Control groups and decreased in all diabetic experimental groups fed on H$_2$O-fraction of DJT and Se supplementation compared with the STZ-Control group. The levels of liver triglyceride were higher in the DJT-Se group than the STZ-Control group. The concentrations of MDA in lung decreased greatly by the administration of Se among all and the concentration of liver MDA was significantly reduced and that of DJT-Se group was the lowest. In conclusion, the results indicated that the administration of H$_2$O-fraction of DJT with selenium supplementation has a synergistic antioxidative effect by influencing on lipid metabolites and peroxidation especially in liver.

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The Effect of Cold Shock on Function and Morphology of Dog Epididymal Spermatozoa (개에서 cold shock이 정소상체유래의 정자의 기능과 형태에 미치는 영향)

  • Yu Il-jeoung
    • Journal of Veterinary Clinics
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    • v.21 no.4
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    • pp.329-335
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    • 2004
  • Dog spermatozoa were recovered from the caudae epididymides of 23 domestic dogs which were 11 pure breed and 12 mix-breed dogs ranging in age from 0.6 to 3 years. The experimental designs were as follows: 1) the effect of chilling to 0. 10 or 37$^{\circ}C$. 2) the kinetics of chilling injury at 0 or 4$^{\circ}C$, and 3) the effect of sugars at $0^{\circ}C$. Viable spermatozoa were recovered by percoll gradient separation and adjusted to 5${\times}$10$^{7}$ spermatozoa/ml. In experiment 1, spermatozoa were diluted with 0.33 M glucose supplemented with 3% BSA (G-BSA) at 1:2 dilution. Spermatozoa were loaded into straws and exposed at 0, 10 or 37$^{\circ}C$ for 30 min. In experiment 2, spermatozoa were prepared as the experiment 1 and exposed for 0.5, 5, 15, or 30 min at 0 or 4$^{\circ}C$. In experiment 3, spermatozoa were diluted with different sugars (0.33 M galactose, glucose, fructose, mannitol, lactose, sucrose, raffinose) and cooled to $0^{\circ}C$ for 30 min. Sperm membrane integrity, motility and acrosome integrity were assayed after rewarming at 37$^{\circ}C$ for 5 min. Sperm motility and membrane integrity abruptly decreased with decreasing temperature but acrosome integrity gradually decreased (P<0.05). Sperm motility was more sensitive to cold shock than membrane integrity and acrosome integrity. Spermatozoa cooled to $0^{\circ}C$ were more damaged than those at 4$^{\circ}C$. Sperm motility was not different among exposed times at both. 0 and 4$^{\circ}C$. However, membrane integrity of spermatozoa exposed for 30 min at both 0 and 4$^{\circ}C$ was significantly lower (P<0.05). Spermatozoa diluted in 0.33 M fructose or galactose showed lower motility and higher morphological abnormality with coiled tail at $0^{\circ}C$. These sperm characteristics were strongly related. These results indicate that dog epididymal spermatozoa are relatively sensitive to rapid cooling and higher morphological abnormality at $0^{\circ}C$ was shown in spermatozoa diluted in fructose and galactose.

Brain Activation Pattern and Functional Connectivity during Convergence Thinking and Chemistry Problem Solving (융합 사고와 화학문제풀이 과정에서의 두뇌 활성 양상과 기능적 연결성)

  • Kwon, Seung-Hyuk;Oh, Jae-Young;Lee, Young-Ji;Eom, Jeung-Tae;Kwon, Yong-Ju
    • Journal of the Korean Chemical Society
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    • v.60 no.3
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    • pp.203-214
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    • 2016
  • The purpose of this study was to investigate brain activation pattern and functional connectivity during convergence thinking based creative problem solving and chemistry problem solving to identify characteristic convergence thinking that is backbone of creative problem solving using functional magnetic resonance imaging(fMRI). A fMRI paradaigm inducing convergence thinking and chemistry problem solving was developed and adjusted on 17 highschool students, and brain activation image during task was analyzed. According to the results, superior frontal gyrus, middle frontal gyrus, inferior frontal gyrus, medial frontal gyrus, cingulate gyrus, precuneus and caudate nucleus body in left hemisphere and cuneus and caudate nucleus body in right hemisphere were significantly activated during convergence thinking. The other hand, middle frontal gyrus, medial frontal gyrus and caudate nucleus in left hemisphere and middle frontal gyrus, lingual gyrus, caudate nucleus, thalamus and culmen of cerebellum in right hemisphere were significantly activated during chemistry problem solving. As results of analysis functional connectivity, all of areas activated during convergence thinking were functionaly connected, whereas scanty connectivity of chemistry problem solving between right middle frontal gyrus, bilateral nucleus caudate tail and culmen. The results show that logical thinking, working memory, planning, imaging, languge based thinking and learning motivation were induced during convergence thinking and these functions and regions were synchronized intimately. Whereas, logical thinking and inducing learning motivation functioning during chemistry problem solving were not synchronized. These results provide concrete information about convergence thinking.