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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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Fertilizer and Organic Inputs Effects on CO2 and CH4 Emission from a Soil under Changing Water Regimes (토양 수분 변동 조건에서 시비 및 유기물 투입에 따른 CO2와 CH4 방출 특성)

  • Lim, Sang-Sun;Choi, Woo-Jung;Kim, Han-Yong
    • Korean Journal of Environmental Agriculture
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    • v.31 no.2
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    • pp.104-112
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    • 2012
  • BACKGROUND: Agricultural inputs (fertilizer and organic inputs) and water conditions can influence $CH_4$ and $CO_2$ emission from agricultural soils. This study was conducted to investigate the effects of agricultural inputs (fertilizer and organic inputs) under changing water regime on $CH_4$ and $CO_2$ emission from a soil in a laboratory incubation experiment. METHODS AND RESULTS: Four treatments were laid out: control without input and three type of agricultural inputs ($(NH_4)_2SO_4$, AS; pig manure compost, PMC; hairy vetch, HV). Fertilizer and organic inputs were mixed with 25 g of soil at 2.75 mg N/25 g soil (equivalent to 110 kg N/ha) in a bottle with septum, and incubated for 60 days. During the first 30-days incubation, the soil was waterlogged (1 cm of water depth) by adding distilled water weekly, and on 30 days of incubation, excess water was discarded then incubated up to 60 days without addition of water. Based on the redox potential, water regime could be classified into wetting (1 to 30 days), transition (31 to 40 days), and drying periods (41 to 60 days). Across the entire period, $CH_4$ and $CO_2$ flux ranged from 0 to 13.8 mg $CH_4$/m/day and from 0.4~1.9 g $CO_2$/m/day, and both were relatively higher in the early wetting period and the boundary between transition and drying periods. During the entire period, % loss of C relative to the initial was highest in HV (16.4%) followed by AS (8.1%), PMC (7.5%), and control (5.4%), indicating readily decomposability of HV. Accordingly, both $CH_4$ and $CO_2$ fluxes were greatest in HV treatment. Meanwhile, the lower $CH_4$ flux in AS and PMC treatments than the control was ascribed to reduction in $CH_4$ generation due to the presence of oxidized compounds such as ${SO_4}^{2-}$, $Fe^{3+}$, $Mn^{4+}$, and ${NO_3}^-$ that compete with precursors of $CH_4$ for electrons. CONCLUSION: Green manure such as HV can replace synthetic fertilizer in terms of N input, however, it may increase $CH_4$ emission from soils. Therefore, co-application of green manure and livestock manure compost needs to be considered in order to achieve satisfactory N supply and to mitigate $CH_4$ and $CO_2$ emission.

Antimicrobial Activity of Sodium chlorate and Korean Herbal Extracts against Mice infected with Escherichia coli O157:H7 (Escherichia coli O157:H7에 감염된 마우스에 대한 염소산나트륨과 한약재 복합추출물 합제의 항균효과)

  • Cha, Chun-Nam;Lee, Yeo-Eun;Son, Song-Ee;Park, Eun-Kee;Choi, Hyun-Ju;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.81-86
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    • 2012
  • The present study was evaluated the antibacterial effect of the combination of $Coptidis$ $rhizoma$, $Glycyrrhiza$ $uralensis$ Fischet, $Schizandra$ $chinensis$ and $Corni$ $Fructus$(1:1:1) extracts(CGSC10). Furthermore, the effectiveness of CGSC10, sodium chlorate, and the combination of CGSC10 and sodium chlorate(CGSCS10) against $E.$ $coli$ O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 5, 10, and 20% CGSC10 was inhibited the growth of $E.$ $coli$ O157:H7 by 34.7, 60.2, and 76.4%, respectively. For 7 days after single challenge with $E.$ $coli$ O157:H7, forty female ICR mice were divided into four experimental groups which were administered in drinking water with saline, 10% CGSC10, 15 mM sodium chlorate, and CGSCS10, respectively. On the 3rd day, the number of $E.$ $coli$ O157:H7 in mouse feces was significantly decreased by administration of CGSC10, 15 mM sodium chlorate, and CGSCS10 ($p$ < 0.001). On the 7th day-after administration, CGSC10, sodium chlorate, and CGSCS10 were decreased the number of $E.$ $coli$ O157:H7 by 27.1, 67.7, and 83.3%, respectively. According to the results of the present study, administration of CGSCS10 to mice can reduce the severity of $E.$ $coli$ O157:H7 infection. In addition, it is suggested that CGSCS10 represents a good candidate for the treatment of enteric infections in domestic animals.

The Egg Development of Korean Slender Gudgeon, Squalidus gracilis majimae (Cypriniforms: Cyprinidae) (한국산 긴몰개 (Squalidus gracilis majime, Cyprinidae)의 난발생)

  • Park, Kyung-Seo;Hong, Young-Pyo;Moon, Woon-Ki;Choi, Shin-Suk;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.38 no.1 s.110
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    • pp.73-82
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    • 2005
  • This study was conducted, based on the field survey and laboratory observations, to elucidate egg developmental processes and their characteristics of the Korean slender gudgeon, Squalidus gracilis majimae. For the experiments, the mature adults were collected at the Woongcheon-Cheon Stream and Boreung Reservoir located in Boreung City, Chungnam Province and eggs were obtained from the natural spawning area. Morphological characteristics of the egg and embryonic development were summarized as follows: The shape of the fertilized egg was spherical, adhesive and transparent. The fertilized egg was 2.9${\pm}$0.3 mm (n = 30) in mean diameter under water temperature of $26{\pm}1.5^{\circ}C$, light white in color and had no oil droplets. After 20 minutes from the time of fertilization, a blastodisc was formed and divided into two cells at 48 minutes after fertilization. The blastular stage occurred at 5 hours 40 minutes after fertilization and the gastrular stage was detected at 8 hours 41 minutes after fertilization. The beginning of embryo formation was observed at 12 hours 58 minutes after fertilization and optic vesicles and 9 somites were discovered at 17 hours 05 minutes after fertilization. Differentiation of brains and embryo wiggling were observed at 37 hours 27 minutes after fertilization. Heart beating and the formation of melanophores in optic vesicles were detected at 44 hours 46 minutes after fertilization. The formation of pectoral fins and melanophores in the body were discovered at 50 hours 36 minutes after fertilization. Hatching occurred at 57 hours 49 minutes after fertilization. The newly hatched larvae were 3.3${\pm}$0.2 mm (n = 120) in total length. We believe that these results may contribute the species and population conservations under the situation of accelerated water pollution and the decreases of its diversity.

Development and validation of an analytical method for nematicide imicyafos determination in agricultural products by HPLC-UVD (HPLC-UVD를 이용한 살선충제 imicyafos의 시험법 개발 및 검증)

  • Do, Jung-Ah;Park, Hyejin;Kwon, Ji-Eun;Choi, Won-Jo;Lee, Hyun-Sook;Chang, Moon-Ik;Hong, Jin-Hwan;Oh, Jae-Ho
    • Analytical Science and Technology
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    • v.27 no.5
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    • pp.234-242
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    • 2014
  • Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.

Bactericidal Efficacy of Fumagari OPP®, Fumigant Against Staphylococcus aureus (훈증소독제, Fumagari OPP®의 Staphylococcus aureus에 대한 살균효과)

  • Cha, Chun-Nam;Park, Eun-Kee;Choi, Hyunju;Kim, Yongpal;Yoo, Chang-Yeol;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.349-353
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    • 2013
  • This study was performed to evaluate the bactericidal efficacy of Fumagari OPP$^{(R)}$, fumigation disinfectant, containing 20% ortho-phenylphenol against Staphylococcus aureus (S. aureus). In this research, efficacy test of fumigant against S. aureus was carried out according to French standard NF T 72-281. S. aureus working culture suspension number (N value), all of the colony numbers on the carriers exposed with the fumigant (n1, n2, and n3), the number of bacterial test suspentions by pour plate method (N1), the number of bacterial test suspentions by filter membrane method (N2) and the mean number of bacteria recovered on the control-carriers (T value) were obtained from the preliminary test. In addition, the reduction number of S. aureus exposed with the fumigant (d value) was calculated using T value, the mean number of bacteria in recovery solution (n'1) and the mean number of bacteria on carriers plated in agar (n'2). N value was $4.0{\times}10^8$ CFU/mL, and n1, n2, and n3 were higher than 0.5N1, 0.5N2 and 0.5N1, respectively. Additionally, T value was $3.4{\times}10^6$ CFU/mL. In the bactericidal effect of the fumigant, the d value was 6.43 logCFU/mL. According to the French standard for the fumigant, the d value for the effective bactericidal fumigant should be over than 5 logCFU/mL. The results indicated that Fumagari OPP$^{(R)}$ had an effective bactericidal activity against S. aureus, then the fumigant can be applied to disinfect food materials and kitchen appliances contaminated with pathogenic bacteria.

GAMETOGENESIS AND EARLY DEVELOPMENT OF LINUPARUS TRIGONUS(VON SIEBOLD) (펄닭새우 생식세포형성과정 및 초기발생)

  • KIM Chang-Hyeon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.2
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    • pp.71-96
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    • 1977
  • Early development Linuparus trigonus(von Siebold) has been studied based on the samples collected monthly in Je-ju Island, Korea from February, 1975 to January, 1977. Gametogenesis, reproductive cycle, embryonic development were investigated by histological mettled, and morphological description was made on the first phyllosoma larva which reared in the laboratory. Testis is composed of two tubular duct which are symmetrical with H-shaped appearance. Outer layer of testis is of fibrous connective tissue capsule. In the lumen there is a convoluted seminiferous tubule with interstitial tissue. Ovary is a pair of symmetrical blind tubular lobes, and the midportions are connected each other. The ovary consists of a couple of ovarian sacs partitioned by two-layered connective tissue fibers. Proliferation of spermatogonia are observed all the year around on the germinal epithelium of seminiferous tubule. Partial spermatogenesis is always in progress, and the spermatozoa appear all the year around in the tubules. Nutrition of early oogonia is supplied by fibrous mesenchyme which is abundantly distributed in ovarian sacs. Oocytes grow and couplete maturation divisions in the follicle layers. They finally develop into mature ova before spawning. Reproductive cycle is classified into four successive stages; multiplication stage from September to December, growing stage from January to March, maturation division stage from April to May and mature stage from June to August. Spawning takes place from May to August with peak spawning from Into July to early August. Cleavage type is superficial. Blastopore is formed in blasto-disc region which is proliferation of blastoderm cells. Germinal layers are also derived from tile region. Mesoderm formation is originated from endodermal cells which are formed front the blasto-disc region. The endodermal cells are separated by the process of delamination from yolk sac and take part in the formation of the mid-gut. Morphological characteristics of first phyllosoma larva are different from the larvae of other Palinurid and Scyllarid species.

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Reproductive Cycle of a Rockfish, Sebastes schlegeli (조피볼락의 생식주기)

  • BAEK Jae-Min;HAN Chang-Hee;KIM Dae-Jung;PARK Chul-Won;Aida Katsumi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.5
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    • pp.431-438
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    • 2000
  • To clarify the annual reproductive cycle in a rockfish, Sebastes schlegeli, monthly changes in gonadosomatic index (GSI), hepatosomatic index (HSI) and histological feature of gonads and plasma levels of sex steroid hormones ($estradiol-l7{\beta},\;17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one,\;testosterone\;and\;11-ketotestosterone$) were investigated. The annual reproductive cycle in females could be divided into 5 periods as follows: 1) recovery period (June to September): serum level of $estradiol-l7{\beta}$ increased gradually; 2) vitellogenesis period (Septemer to february) : vitellogenic oocytes were obsewed, GSI sustained high value, and serum level of $estradiol-l7{\beta}$ increased; 3) gestation period (February-April): developing larva showed in the ovary, and serum levels of $17{\alpha},\;20{\beta}-dihydroxy-4-pregnen-3-one$ and testosterone increased; 4) partrition period (April to May) : larva were delivered, and value of GSI and serum levels of hormones decreased rapidly; 5) resting period (May to June) : value of GSI and serum levels of $estradiol-l7{\beta}$ and testosterone remained low. The annual reproductive cycle in males could be divided into 6 periods; 1) early maturation period (April to June): value of GSI and serum levels of hormones incresed gradually, cyst of spermatogonia incresed in number, and a small number of cyst of spermatocyte was observed; 2) mid-maturation perid (June to September); value of GSI and serum levels of hormones increased, and germ cells in many cysts were undergoing active sperrnatogenesis; 3) late maturation period (September to November) : value of GSI and serum levels of hormones remained high and spermatozoa were released into the lumina of the seminal lobules; 3) spermatozoa dischaging period (Nobember to December) : the lumina of the seminal lobules were enlarged and filled with mature spermatozoa; 4) degeneration period (December to Februauy)i value of GSI decresed and cyst of spermatocyte were decresed in number; 5) resting period (December to April) : no histological changes of testes were observed, and value of GSI and serum levels of hormones remained low. In November, the lumina of the seminal lobules were filled with mature spermatozoa and sperm masses were present in the ovarian cavity. Thus, copulation in this species occurred in November and December.

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The apoptotic fragment assay in rat peripheral lymphocytes and crypt cells with whole body irradiation with 60Co ϒ-rays and 50 MeV cyclotron fast neutrons (코발트-60 감마선과 50 MeV 싸이크로트론 고속 중성자선에 전신조사된 랫드의 말초 임파구와 음와 세포의 아포토시스 유도를 이용한 생물학적 선량 측정 모델 개발 연구)

  • Kim, Tae-hwan
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.203-210
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    • 2001
  • Here, we compared the effectiveness of 50 MeV($p{\to}RBe^+$) cyclotron fast neutrons versus $^{60}Co$ ${\gamma}$-rays by the apoptotic fragment frequency in both rat peripheral lymphocytes and crypt cells to check a radiobiological endpoint. The incidence of apoptotic cell death was increased in all irradiated groups, and radiation at all doses trigger rapid changes in both crypt cells and peripheral lymphocytes. These data suggest that apoptosis may play an important role in homeostasis of damaged radiosensitive target organ by removing damaged cells. The curve of dose-effect relationship for these data of apoptotic fragments frequencies was $y=0.3+(6.512{\pm}0.279)D(r^2=0.975)$ after neutrons, while $y=0.3+(4.435{\pm}0.473)D+(-1.300{\pm}0.551)D^2(r^2=0.988)$ after ${\gamma}$-rays. In addition, $y=3.5+(118.410{\pm}10.325)D+(-33.548{\pm}12.023)D^2(r^2=0.992)$ after ${\gamma}$-rays in rat lymphocytes. A significant dose-response relationship was found between the frequency of apoptotic cell and dose. These data show a trend towards increase of the numbers of apoptotic cells with increasing dose. Dose-response curves for high and low linear energy transfer (LET) radiation modalities in these studies were different. The relative biological effectiveness (RBE) value for crypt cells was 1.919. In addition, there were significant peaks on apoptosis induction at 4 and 6h after irradiation, and the morphological findings of the irradiated groups were typical apoptotic fragments in crypt cells that were hardly observed in the control group. Thus, apoptosis induction in both crypt cells and peripheral lymphocytes could be a useful endpoint of rat model for studying screening test and microdosimetic indicator to evaluate the biological effects of radiation-induced cell damage.

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Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.