Purification and characterization of a novel fibrinolytic α chymotrypsin like serine metalloprotease from the edible mushroom, Lyophyllum shimeji

Moon, S.M.,Kim, J.S.,Kim, H.J.,Choi, M.S.,Park, B.R.,Kim, S.G.,Ahn, H.,Chun, H.S.,Shin, Y.K.,Kim, J.J.,Kim, D.K.,Lee, S.Y.,Seo, Y.W.,Kim, Y.H.,Kim, C.S. Society for Bioscience and Bioengineering, Japan ; 2014 Journal of bioscience and bioengineering Vol.117 No.5

A novel fibrinolytic enzyme was purified from Lyophyllum shimeji, a popular edible mushroom in Asia. The enzyme was purified using combination of anion exchange chromatography on a Mono Q 5/5 column and size exclusion gel filtration chromatography on Superdex 200 100/300 column. This purification protocol resulted 80.9-fold purification of the enzyme and a final yield of 5.7%. The molecular weight of the purified enzyme was estimated to be 21 kDa by SDS-PAGE and size exclusion gel filtration. The N-terminal amino acid sequence was found to be ITFQSASP, which is dissimilar from that of known fibrinolytic enzymes. The purified enzyme was a neutral protease with an optimal reaction pH and temperature of 8.0 and 37<SUP>o</SUP>C, respectively. Enzymatic activity was inhibited by Cu<SUP>2+</SUP> and Co<SUP>2+</SUP>. It was also significantly inhibited by PMSF and TPCK. Furthermore, it was found to exhibit a higher specificity for S-7388, a well-known chymotrypsin chromogenic substrate, indicating chymotrypsin like serine metalloprotease. The relative fibrinolytic activity of 5 μg purified enzyme have two fold more activity than 1 unit/ml of plasmin on fibrin plate. Furthermore, purified enzyme preferentially hydrolyzed the Aα-chain followed by the Bβ- and γ-chain of fibrinogen, which is precursor of fibrin. Therefore, these data suggests that the fibrinolytic enzyme derived from edible mushroom, L. shimeji, might be useful for thrombolytic therapy and preventing thrombotic disease.

Changes in spatial variations of sap flow in Korean pine trees due to environmental factors and their effects on estimates of stand transpiration

Moon, M.,Kim, T.,Park, J.,Cho, S.,Ryu, D.,Suh, S.,Kim, H. S. Springer Science + Business Media 2016 Journal of mountain science Vol.13 No.6

<P>It is difficult to scale up measurements of the sap flux density (J(S)) for the characterization of tree or stand transpiration (E) due to spatial variations in J(S) and their temporal changes. To assess spatial variations in the sap flux density of Korean pine (Pinus koraiensis) and their effects on E estimates, we measured the J(S) using Granier-type sensors. Within trees, the J(S) decreased exponentially with the radial depth, and the J(S) of the east aspects were higher than those of the west aspects. Among trees, there was a positive relationship between J(S) and the tree diameter at breast height, and this positive relationship became stronger as the transpiration demand increased. The spatial variations that caused large errors in E estimates (i.e., up to 110.8 % when radial variation was ignored) had varied systematically with environmental factors systematic characteristics in relation to environmental factors. However, changes in these variations did not generate substantial errors in the E estimates. For our study periods, the differences in the daily E (E-D) calculated by ignoring radial, azimuthal and tree-to-tree variations and the measured ED were fairly constant, especially when the daily vapor pressure deficit (D_(D)) was higher than 0.6 kPa. These results imply that the effect of spatial variations changes on sap flow can be a minor source of error compared with spatial variations (radial, azimuthal and tree-to-tree variations) when considering E estimates.</P>

난관배양액이 처녀발생유기된 돼지난포란의 체외발달에 미치는 영향

문승주,이경호,김호,김창렬,은대숙,김광현,나진수,김재홍 全南大學校 農業科學技術硏究所 1997 農業科學技術硏究 Vol.32 No.-

본 연구는 난관배양액이 돼지수정란의 체외발달에 미치는 효과를 규명키 위하여 수행하였다. 돼지 미성숙 난포란은 TCM-199, Ham's F-10 그리고 Whitten's 배양액에 10% 난포액과 호르몬(PMSG : 10IU/㎖, HCG : 10IU/㎖)을 첨가 20시간 배양하고 호르몬을 첨가하지 않는 배양액에서 20시간 추가 배양하여 총 40시간동안 배양하여 체외성숙을 유도하였다. 체외성숙후 0.1% hyaluronidase로 난구세포를 제거하고 15% FCS가 함유된 TCM-199으로 3회 세척하고 TCM-199에 15% FCS와 10% ethanol 혼합액에 세척한 난자를 옮겨 10분간 배양 처녀발생을 유기하였다. 처녀발생 6시간후 전핵형성율은 체외성숙배양액으로 TCM-199을 사용했을 때 56.4%, Ham's F-10의 경우 58.3%, Whitten's 배양액의 경우 74.0%를 보였다. 처녀발생 유기 48시간째 난할율은 TCM-199을 사용했을 때 45.7%, Ham's F-10에서 45.4%, Whitten's배양액에서 39.2%를 보였으며 세종류의 배양액에 POCM을 첨가 배양했을 때 TCM-199에 44.8%, Ham's F-10에서 45.4%, Whitten's배양액에서 43.7%로 나타났다. 처녀발생육 난자를 96시간 체외배양시킨 결과 상실배 발달율이 POCM을 첨가 했을 때 첨가하지 않은 시험구에 비하여 유의적으로 높았다(P<0.05) The effect of porcine oviductal conditioned medium(POCM) on in vitro development of chemically activated porcine oocytes was studied. Porcine oocytes were cultured in TCM-199, Ham's F-10 and Whitten's medium with hormonal supplements for 20h and 40h additional culture without hormonal supplements. After in vitro maturation, the denuded oocytes were washed 3 times with TCM-199 contaning 15%(v/v) ethanol to induce pathenogenetic activation. At 6h after activation, pronuclea formation rates were 56.4% in TCM-199, 59.3% in Ham's F-10 and 74.0% in Whitten's maturation medium. At 48h after activation, 45.7%, 45.4% and 39.2% of oocytes claved in TCM-199, Ham's F-10 and Whitten's culture medium, respectively. And 44.8%, 45.5% and 43.7% of oocytes were claved in TCM-199, Ham;s F-10 and Whitten's culture medium supplemented with POCM, respectively. The rates of moular were higher in culture medium with POCM than without POCM at 96h after activation.(P<0.05)

유전자 알고리즘을 이용한 InGaP/GaAs HBT 소신호 등가회로 파라미터 추출

장덕성(D.S. Chang),문종섭(J.S. Moon),박철순(C.S. Park),윤경식(K.S. Yoon) 한국지능시스템학회 2001 한국지능시스템학회논문지 Vol.11 No.6

에미터 크기가 2×10 μm²인 InGaP/GaAs 이종접합 바이폴라 트랜지스터의 T자 모양으로 연결된 등가회로 요소를 추출하기 위하여, 경계 구간 설정이 개선된 유전자 알고리즘을 채택하였다. 이 소신호 모델 파라미터를 유전자 알고리즘을 사용하여, 다양한 순방향 바이어스에서 측정한 S-파라미터들로부터 추출하였다. 추출된 값들은 물리적인 의미와 일관성을 보여준다. 모델 S-파라미터는 측정 S-파라미터와 2GHz- 26.5GHz의 주파수 범위에서 잘 일치한다. The present approach based on the genetic algorithm with improved selections of bounds was adopted to extract a bridged T equivalent circuit elements of a 2×10 μm² InGaP/GaAs HBT. The small-signal model parameters were extracted using the genetic algorithm from S-parameters measured at different frequencies under multiple forward-active biases, which demonstrate physically meaningful values and consistency. The agreement between the measured and modeled S-parameters is excellent over the frequency range of 2 to 26.5GHz.

miR-302b maintains ''stemness'' of human embryonal carcinoma cells by post-transcriptional regulation of Cyclin D2 expression

Lee, N.S.,Kim, J.S.,Cho, W.J.,Lee, M.R.,Steiner, R.,Gompers, A.,Ling, D.,Zhang, J.,Strom, P.,Behlke, M.,Moon, S.H.,Salvaterra, P.M.,Jove, R.,Kim, K.S. Academic Press 2008 Biochemical and biophysical research communication Vol.377 No.2

Embryonic stem cells (ESCs) and embryonal carcinoma cells (ECCs) possess the remarkable property of self-renewal and differentiation potency. They are model preparations for investigating the underlying mechanisms of ''stemness''. microRNAs are recently discovered small noncoding RNAs with a broad spectrum of functions, especially in control of development. Here, we show that miR-302b indirectly regulates expression of the pluripotent stem cell marker Oct4, and it directly regulates expression of Cyclin D2 protein, a developmental regulator during gastrulation. Using loss-of function and gain-of function approaches, we demonstrate that functional miR-302b is necessary to maintain stem cell self-renewal and inhibit neuronal differentiation of human ECCs. During retinoic acid-induced neuronal differentiation, Cyclin D2 protein but not mRNA expression is strongly increased, concurrent with the down-regulation of miR-302b and Oct4. Our results suggest that miR-302b plays an important role in maintaining the pluripotency of ECCs and probably ESCs, by post-transcriptional regulation of Cyclin D2 expression.

Prevalence and characterization of Salmonella in pigs from conventional and organic farms and first report of S. serovar 1,4,[5],12:i:- from Korea

Tamang, M.D.,Gurung, M.,Nam, H.M.,Moon, D.C.,Kim, S.R.,Jang, G.C.,Jung, D.Y.,Jung, S.C.,Park, Y.H.,Lim, S.K. Elsevier Scientific Pub. Co 2015 Veterinary microbiology Vol.178 No.1

<P>This study compared the prevalence of Salmonella spp. and their antimicrobial susceptibilities in pigs from conventional and organic farms during 2012-2013 in Korea and characterized them by molecular methods. Altogether, 100 nontyphoid Salmonella were isolated: 47 from 1324 pigs (3.5%) from conventional farms and 53 from 641 pigs (8.3%) from organic farms. The most frequent serovar was Typhimurium (49%) followed by Panama (24%), 1,4,[5],12:i:- (5%), and Virchow (5%). Overall, the isolates were most often resistant to tetracycline (75%) followed by ampicillin (66%), streptomycin (57%), and gentamicin (44%). The prevalence of antimicrobial resistance, multi-drug resistance phenotype, and resistance to tetracycline, ampicillin, and gentamicin were significantly higher in swine Salmonella from conventional farms than those from organic farms. The most common resistance pattern was ampicillin-gentamicin-tetracycline (n = 16). All eight ceftiofur-resistant Salmonella identified produced CTX-M-15. Overall, decreased susceptibility to ciprofloxacin was observed in 39 isolates. Among them, a single isolate was positive for qnrS1 gene. An insertion sequence ISEcp1 was detected upstream of bla(CTX-M) gene in all isolates. The spread of bla(CTX-M-15) gene was attributed to combination of clonal expansion and horizontal dissemination mediated by IncHI2 plasmid. Multilocus variable number of tandem repeats analysis demonstrated clonal dissemination of S. Typhimurium and S. 1,4,[5],12:i:- strains in pigs. To our knowledge, this is the first report of bla(CTX-M-15) gene in S. Virchow from pigs and qnrS1 gene in S. Rissen from animals. This study also reports the first occurrence of Salmonella serovar 1,4,[5],12:i:- from Korea and CTX-M-15 producing Salmonella from pigs in Korea. (C) 2015 Elsevier B.V. All rights reserved.</P>

두개골의 단순전후 방사선 사진을 이용한 전두동의 분류 및 그 분포에 관한 연구

이혜경,권귀향,김기정,강신몽,문국진 순천향대학교 2002 Journal of Soonchunhyang Medical Science Vol.8 No.4

Cowden's Disease - A Report on the First Case in Korea and Literature Review

Lee, H.-R Korean Academy of Medical Science 1997 Journal of Korean medical science Vol.12 No.6

Vitamin D-binding protein interacts with Aβ and suppresses Aβ-mediated pathology

Moon, M,Song, H,Hong, H J,Nam, D W,Cha, M-Y,Oh, M S,Yu, J,Ryu, H,Mook-Jung, I Macmillan Publishers Limited 2013 CELL DEATH AND DIFFERENTIATION Vol.20 No.4

The level of vitamin D-binding protein (DBP) is increased in the cerebrospinal fluid of patients with Alzheimer’s disease (AD), suggesting a relationship with its pathogenesis. In this study, we investigated whether and how DBP is related to AD using several different approaches. A pull-down assay and a surface plasmon resonance binding assay indicated direct interactions between purified DBP and amyloid beta (Aβ), which was confirmed in the brain of AD patients and transgenic AD model mice by immunoprecipitation assay and immunohistochemical double-staining method. Moreover, atomic force microscopic examination revealed that DBP reduced Aβ aggregation in vitro. DBP also prevented Aβ-mediated death in cultured mouse hippocampal HT22 cell line. Finally, DBP decreased Aβ-induced synaptic loss in the hippocampus and rescued memory deficits in mice after injection of Aβ into the lateral ventricle. These results provide converging evidence that DBP attenuates the harmful effects of Aβ by a direct interaction, and suggest that DBP is a promising therapeutic agent for the treatment of AD.

Impact of White Matter Changes on Activities of Daily Living in Mild to Moderate Dementia

Moon, So Young,Na, Duk L.,Seo, Sang Won,Lee, Jun-Young,Ku, Bon D.,Kim, Seong Yoon,Park, Kyung Won,Shim, Yong S.,Youn, Young Chul,Chung, Chan-Seung,Cheong, Hae-Kwan,Choi, Seong Hye,Cha, Kyung Ryeol,Kim S. Karger AG 2011 European neurology Vol.65 No.4

<P>The association between white matter changes and activities of daily living (ADL) in a large, well-defined cohort of patients with mild-to-moderate dementia (either Alzheimer’s disease or subcortical vascular dementia) were investigated. A total of 289 patients were divided into three groups (140 mild, 99 moderate, and 50 severe) depending on the degree of white matter changes as indicated on brain magnetic resonance image scans. Further, we analyzed the three groups’ performances on basic and instrumental ADL. The degree of white matter changes was associated with greater age, hypertension, previous history of stroke, higher Hachinski Ischemic Score, worse global cognitive and functional status, and an increased impairment of basic ADL and instrumental ADL. The increased impairment with regard to the severe group’s performance on both the basic and instrumental ADL remained significant after adjustment for age and hypertension. Tasks involving physical activities were most significant. This was the first study investigating the association between white matter changes and ADL in a large, well-defined dementia cohort. The present study suggests that severe white matter changes may be associated with higher impairment on both basic and instrumental ADL.</P><P>Copyright © 2010 S. Karger AG, Basel</P>