• Title/Summary/Keyword: Y-protein

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Characterization of an Antarctic alkaline protease, a cold-active enzyme for laundry detergents (세탁세제 첨가용 효소 개발을 위한 남극 해양세균 유래 저온성 단백질분해효소의 특성 연구)

  • Park, Ha Ju;Han, Se Jong;Yim, Joung Han;Kim, Dockyu
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.60-68
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    • 2018
  • A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

Analysis of antigen specificity using monoclonal and polyclonal antibodies to cysticercus cellulosae by enzyme-linked immunoelectrotransfer blot technique (효소면역전기영동이적법을 이용한 유조설고충 단세후군항체 및 환기혈청에 대한 항원특리성 분석)

  • Jo, Seung-Yeol;Gang, Sin-Yeong;Kim, Seok-Il
    • Parasites, Hosts and Diseases
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    • v.25 no.2
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    • pp.159-167
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    • 1987
  • To analyse the antigen specificity of patients sera from 24 confirmed neurocysticercosis and a monoclonal antibody, SDS-PAGE using 10~15% linear gradient gel and EITB were done. Cystic fluid, saline extracts of scolex and of whole worm of C. cellulosae, saline extracts of sparganum, hydatid cyst fluid, saline extracts of Fasciola, Clonorchis and Paragonimus were used as antigen. Of protein bands in cystic fluid of C. cellulosae, patient sera reacted frequently to bands of 152, 94, 64, 48, 24, 15, 10 and 7kDa proteins. To saline extracts of scolex and whole worm of C. cellulosae, patients sera reacted frequently to 94, 64, 52, 39, 34, 15 and 10kDa bands. Two bands in sparganum extract (130 and 64kDa) and two bands in hydatid cyst fluid (52 and 27kDa) were cross-reacting bands with sera from cysticercosis patients. Saline extracts of Fasciola, ClonorchiJ and Paragonimus did 'not exhibit cross-reacting bands. Monoclonal antibody to cystic fluid of C. cellulosae was found to react with low molecular weight proteins of 15, 10 and 7kDa.

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Antigenic localities in the tissued of Metagonimus yokogawai observed by immunogoldlabeling method (면역황금 표식법을 이용한 요꼬가와흡충의 조직내 항원성 부위에 관한 연구)

  • Ahn, Hyuk;Rim, Han-Jong;Kim, Soo-Jin
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.245-258
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    • 1991
  • In order to determine the antigenic localization in the tissues of the adult Metagonimus yokegawai, immunogoldlabeling method was applied using serum immunoglobulins (IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size: 12 nm) , It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.

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Clinical Presentations and Causative Organisms in Children and Adolescents with Osteoarticular Infections: A Retrospective Study (소아청소년기에 발생한 골관절염의 임상 양상 및 원인균에 대한 고찰)

  • Lee, Soyoung;Kim, Han Wool;Cho, Hye-Kyung;Yun, Yoe Hon;Ryu, Kyung Ha;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.22 no.3
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    • pp.154-163
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    • 2015
  • Purpose: Osteoarticular infections in children and adolescents are important because it can cause functional compromise if appropriate treatment is delayed. Therefore, this study was designed to describe the clinical presentations and causative organisms of osteoarticular infections in children and adolescents in order to propose early diagnosis method and an appropriate empiric antimicrobial therapy. Methods: Forty-two medical records were reviewed retrospectively, which were confirmed as osteomyelitis (OM) or septic arthritis (SA) at Department of Pediatrics or Orthopedic Surgery in patients under 18 years old of Ewha Womans University Mokdong Hospital from March 2008 to March 2015. Results: We identified 21 cases of OM, 13 cases of SA and 8 cases of OM with SA. There were 31 males and 11 females and mean age was 7.1 years old. The most common symptoms were pain and tenderness of involved site. Major involved bones were femur (10 cases, 34.5%), tibia (7 cases, 24.1%) and major involved joints were hip (9 cases, 42.9%), and knee (5 cases, 23.8%). Increased serum C-reactive protein and erythrocyte sedimentation rate were observed in 37 cases (88.1%) respectively. Magnetic resonance imaging was performed in 40 cases among 42 cases and was used to demonstrate osteoarticular infections and other adjacent infections. Nine cases (23.7%) among 38 cases and 20 cases (50.0%) among 40 cases were positive in blood culture and infected site culture respectively. The most common causative organism was Staphylococcus aureus, which was represented in 22 cases (75.9%), of which nine cases (40.9%) were resistant to methicillin. Conclusions: S. aureus was the most common causative organism of osteoarticular infections in children and adolescents and the proportion of MRSA was high in this study. Therefore, we recommend vancomycin as the first empiric antimicrobial therapy and suggest that further study is necessary to elucidate an appropriate guideline for treatment which takes into account MRSA proportion.

A Nutrition Intakes Survey of Pregnant Women in a Urban Area -Application of Convenient Method for the Study of Nutritionial Status- (일개 도시지역 임신부의 영양섭취에 관한 조사연구 -간이식 영양조사법 이용-)

  • Kim, In-Sook
    • Journal of Preventive Medicine and Public Health
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    • v.16 no.1
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    • pp.99-104
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    • 1983
  • Aiming at the total 200 pregnant women including 150 ones registered with the Health Center of Chung Ku District and 50 ones registered with the Health Center of Mapo District, we have conducted a research study of the socio-medical characteristics, maternal child health, and the status of nutritional intake which has utilized the application of convenient method for the study of nutritional status, during the period of April 20 to May 25, 1983. And we have obtained the following conclusions: 1. Regarding age distribution, the pregnant women aged from 26 to 30 were most numerous, which was 54.0%. The percentage of women who experienced the first pregnancy was 12.0% and the percentage of those who experienced the second pregnancy was 37.0%, which was the highest. Regarding the weeks of pregnancy of the pregnant women, the first trimesterr was 11.5%, the second trimester 30.0%, and the third trimester 58.6%. 2. Regarding academic achievements, the pregnant women who graduated from middle schools reached 43.5%, which was the highest percentage. Regarding economic status, the pregnant women who owned their own houses were only 21.0%. And the pregnant women whose monthly income was from 300,000 won to 400,000 won were 40%, which was the most numerous. 5. The women above 15 years old who experienced the first menstruation were 84.0%. And those who experienced abortion were totally 54.4%. and 35.5% among those women experienced artificial abortion. 4. 70.5% of the pregnant women said that their health condition was excellent, 24.5% felt subjective complaints, and 5.0% specially received medical consultation for their diseases. 5. 82.0% received prenatal care, but 60.5% regularly received prenatal care. 68.0% received the education for nutrition and only 19.5% regularly received the education for nutrition. 6. Regarding the family composition, the families consisting of two generations were 47.0%, which was the most numerous. 97.5% of the preparation for meals was conducted by housewives. They said that they did not lack time for meal preparation. 7. 94.9% of the pregnant women said that they had eaten as in ordinary times during their pregnancy. 25.5% said that there were tabooed foods. Tabooed foods are chiefly pork, chicken, milk, and eggs. 68.0% don't drink milk during pregnancy, 32.5% take the intake of vitamins, and 20.5% take iron supplement. 8. The average amounts of the intake of protein, fat, and carbohydrate of a pregnant women are 49.3gm, 29.4gm, and 205.1gm respectively, which showed the phenomenon in which the amount of the intake of nutrition increased as the weeks of pregnancy increased. The average amount of the intake of salt was 14.2gm. 9. Regarding the hemoglobin value of all pregnant women, those whose hemoglobin value was less than 11.0gm were 66.5%, those whose hemoglobin value was 11 to 12 gm were 16.5%. and those whose hemoglobin value was above 12gm were 17.0%. The pregnant women whose hemoglobin value was less than 11.0gm in the first trimester of pregnancy, in the second trimester, and in the trimester were 81.8%, 62.8%, and 64.9% respectively. This shows that the phenomenon of anemia increased as the weeks of pregnancy increased 10. Regarding physical development of a pregnant woman during prenatal period, the height and weight were $156.7cm{\pm}14.1$ and $51.1kg{\pm}58$ respectively. When the standard increase of a prenatal weigt gain is set as 100%, the women over the range of 100% were 28.0% and the women under the range of 80% were 37.0%.

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Clinical Significance and Incidence of Gram-positive Uropathogens in Pediatric Patients Younger than 1 Year of Age with Febrile Urinary Tract Infection (1세 이하의 발열성 소아 요로감염에서 Gram-Positive Uropathogens의 발생 빈도 및 임상적 의의)

  • Yang, Tae Hwan;Yim, Hyung Eun;Yoo, Kee Hwan
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.65-72
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    • 2013
  • Purpose: Urinary tract infection (UTI) caused by gram-positive uropathogens is usually hospital-acquired and associated with predisposing conditions. However, the incidence of gram-positive bacteria in community-acquired UTIs has recently increased worldwide. We aimed to investigate the clinical significance of UTI and associated genitourinary malformations in young children with febrile UTIs caused by gram-positive bacteria. Methods: We retrospectively reviewed the medical records of 566 patients (age, <1 year) who visited the Korea University Medical Center for febrile UTIs between January 2008 and May 2013. We classified the patients into the following two groups: gram-positive (P group) and gram-negative (N group), according to the results of urine culture. The fever duration; white blood cell (WBC) counts and C-reactive protein (CRP) levels in peripheral blood; and the presence of hydronephrosis, cortical defects, vesicoureteral reflux (VUR), and renal scarring were compared between the two groups. Results: The number of patients with gram-positive bacteria was 23 (4.1%) and with gram-negative bacteria was 543 (95.9%). The most common pathogen was Escherichia coli, and Enterococcus faecalis showed the highest incidence among gram-positive uropathogens. Patients with gram-positive bacteria showed longer fever duration compared to that in patients with gram-negative bacteria (P vs. N, $3.4{\pm}1.2$ vs. $2.9{\pm}1.6$ days, P <0.05). The incidence of VUR was increased in the gram-positive group compared to that in the gram-negative group (P vs. N, 55.6 vs. 17.8%, P<0.05). However, there were no significant differences in other laboratory and radiologic findings. Conclusion: The findings of our study show that community-acquired UTIs in patients younger than 1 year of age, caused by gram-positive uropathogens, can be associated with prolonged fever duration and the presence of VUR.

Differences in the Clinical Characteristics of Children with Urinary Tract Infections Based on the Results of $^{99m}Tc$-Dimercaptosuccinic Acid Renal Scanning (요로감염 소아에서 입원 초기 시행한 DMSA 신 스캔 결과에 따른 임상양상의 차이에 대한 연구: DMSA 신 스캔의 임상적 의미)

  • Kim, Dong Ouk;Lee, Sang Min;Lee, Jeong Bong;Ko, Young Bin;Kim, Su Jin
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.110-116
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    • 2013
  • Purpose: The $^{99m}Tc$-Dimercaptosuccinic acid (DMSA) renal scan is used primarily for the diagnosis of renal scarring and acute pyelonephritis in children with urinary tract infections (UTI). This study aimed to evaluate clinical differences based on the positive or negative results of DMSA scans and kidney ultrasonography (US) in pediatric UTI. Method: We retrospectively reviewed 142 pediatric patients with UTI who were admitted to Myongji Hospital from January 2004 to December 2012. We performed a comparative analysis of clinical parameters such as age, sex, white blood cell (WBC) count, neutrophil count, blood urea nitrogen (BUN) level, creatinine (Cr) level, C-reactive protein (CRP) level, and durations of hospitalization and fever, grouped by the results of the DMSA scans and kidney US. Results: The mean age of the patients was $33.8{\pm}48.3$ months, and 78 (55%) were male. Fifty-two patients had abnormal DMSA findings, and 71 patients had abormal kidney US findings (test positive groups). In the DMSA scan positive group, there were significant differences in age, WBC counts, neutrophil counts, CRP level, BUN level, Cr level, hospitalization duration, number of abnormal findings on kidney US, and incidence of vesicoureteral reflux (VUR) compared with the scan negative group. The kidney US positive group had significant differences in age, neutrophil count, CRP level, BUN level, Cr level, hospitalization duration, number of abnormal findings on the DMSA scans, and more frequent VUR compared with the US negative group. Conclusion: Our data suggest that there were no major differences in clinical parameters based on the results of the DMSA scans compared with kidney US in pediatric UTI. However, as kidney US and DMSA scan were performed to predict VUR, the sensitivity and negative predictive value was increased.

Clinicopathologic Changes in Children with Immunoglobulin A Nephritis and Henoch-Sch$\ddot{o}$nlein Purpura Nephritis after Cyclosporine A and Angiotensin-converting Enzyme Inhibitor Treatment (Immunuglobulin A 신질환과 Henoch-Schnlein purpura 신질환을 가진 소아에서의 cyclosporine A와 angiotensin-converting enzyme inhibitor 치료의 임상적, 병리학적 변화)

  • Lee, Jeong Ju;Kim, Yong-Jin;Shin, Jae Il;Yim, Hyunee;Park, Se Jin
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.92-100
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    • 2013
  • Purpose: To investigate the clinicopathologic effects of cyclosporine A (CsA) in children with diseases characterized by mesangial immunoglobulin A deposits such as immunoglobulin A nephropathy (IgAN) and Henoch-Sch$\ddot{o}$nlein purpura nephritis (HSPN). Methods: We retrospectively reviewed the clinicopathologic outcomes of 54 children (IgAN, 36; HSPN, 18) treated with CsA. The starting dose of CsA was 5 mg/kg per day, and it was administered in 2 divided doses. The degree of proteinuria and pathologic changes in renal biopsies were evaluated before and after CsA treatment. Results: The mean protein to creatinine ratio decreased from $3.7{\pm}1.5$ to $0.6{\pm}0.4$(P <0.001), and 32 (59.2%) children achieved complete remission of proteinuria after 1-year CsA treatment. Among the 54 children, 24 maintained normal renal function and 25 exhibited microscopic hematuria or proteinuria at the end of CsA treatment. In the HSPN group, 3 children whose initial biopsies indicated class IIIb disease showed class II disease on follow-up, and the follow-up biopsies of 2 children who had class II disease indicated the same class II disease. In the IgAN group, cortical tubular atrophy occurred in 1 child, and no child with IgAN had cortical interstitial fibrosis or tubular atrophy after 1-year CsA treatment. No significant complications were found in the children treated with CsA. Conclusion: Our findings indicate that CsA treatment is effective and beneficial in reducing massive proteinuria and preventing progression to end-stage renal failure in children with glomerular diseases characterized by IgA deposits, such as IgAN and HSPN, within 1 year of treatment.

Extract from Eucheuma cottonii Induces Apoptotic Cell Death on Human Osteosarcoma Saos-2 Cells via Caspase Cascade Apoptosis Pathway (Eucheuma cottonii 추출물에 의한 인체 골육종암 Saos-2 세포의 자가사멸 유도)

  • Kang, Chang-Won;Kang, Min-Jae;Kim, Kyong Rok;Kim, Nan-Hee;Seo, Yong Bae;Kang, Keon-Hee;Kim, Sang-Ho;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.2
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    • pp.147-154
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    • 2016
  • Osteosarcoma (OS) is the most common and malignant bone tumors. Although many types of resection surgery and experimental agents were developed, median survival and clinical prognosis are poorly investigated. Recently, several researches have reported that Eucheuma cottonii has potent as protective effects of coal dust-induced lung damage via inhibition of malondialdehyde (MDA) and oxidative stress in bronchoalveolar lavage fluids (BALF). However, anti-cancer effects and specific molecular mechanism of extract from Eucheuma cottonii (EE) has not been clearly studied yet. This study evaluated that anti-cancer potential of EE in human osteosarcoma Saos-2 cells. EE indicated cytotoxicity on Saos-2 cells in a dose-dependent manner. Morphological degradation and nucleic condensation were also observed under the EE treatment. However, it did not significantly affect on non-cancerous kidney HEK-293 cells under the same concentration which is shown cytotoxicity on Saos-2 cells. The phosphorylation of Fas-Associated Death Domain (FADD) and expression of cleaved caspase-8, -7 and -3 were upregulated in a dose-dependent manner. In immunofluorescence staining, expression level of Fas and cleaved PARP were upregulated by EE treatment. Furthermore, treatment of EE induces upregulation of sub G1 phase by flow cytometry analysis. The results demonstrated that EE has a therapeutic potential against osteosarcoma via FADD mediated caspase cascade apoptosis signal pathway.

Insulin-Like Growth Factor-I Induces Androgen Receptor Coactivator Expression in Skeletal Muscle Cells through the p38 MAPK and ERK1/2 Pathways (C2C12 세포에서 insulin-like growth factor-I이 p38 MAPK, ERK1/2 신호전달 경로를 통해 엔드로젠 수용체 coactivator 발현에 미치는 영향)

  • Park, Chan-Ho;Kim, Hye-Jin;Kim, Tae-Un;Lee, Won-Jun
    • Journal of Life Science
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    • v.21 no.2
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    • pp.242-250
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    • 2011
  • Although insulin-like growth factor-I (IGF-I) and androgen receptor (AR) coactivators are well known effectors of skeletal muscle, the molecular mechanism by which signaling pathways integrating AR coactivators and IGF-I in skeletal muscle cells has not been previously examined. In this study, the effects of IGF-I treatment on the gene expression of AR coactivators in the absence of AR ligands and the roles of the p38 MAPK and ERK1/2 signaling pathways in IGF-I-induced AR coactivators induction were examined. C2C12 cells were treated with 250 ng/ml of IGF-I in the presence or absence of specific inhibitors p38 MAPK (SB203580) or ERK1/2 (PD98059). Treatment of C2C12 cells with IGF-I resulted in increased in GRIP-1, SRC-1, and ARA70 protein expression. The levels of GRIP-1, SRC-1, and ARA70 mRNA were also significantly increased after 5min of IGF-I treatment. IGF-I-induced AR coactivator proteins were significantly blocked by pharmacological inhibitors of p38 MAPK and ERK1/2 pathways. However, there was no significant effect of those inhibitors on IGF-I-induced mRNA level of AR coactivators, suggesting that AR coactivators are post-transcriptionally regulated by IGF-I. Furthermore, the present results suggest that IGF-I stimulates the expression of AR coactivators by cooperative activation of the p38 MAPK and ERK1/2 pathways in C2C12 mouse skeletal muscle cells.