• Title/Summary/Keyword: endo type

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Purification and Properties of ${\beta}-Mannanases$ from Germinated Guar Bean (${\beta}-Mannanase$ 군(群)의 정제(精製) 및 그들의 성질(性質)에 관(關)한 연구(硏究))

  • Lee, Su-Rae
    • Applied Biological Chemistry
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    • v.7
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    • pp.1-13
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    • 1966
  • 1) Three ${\beta}-1$, 4-mannanases were isolated from germinated guar bean through extraction, ammonium sulfate fractionation, column chromatography on cellulose derivatives and gel filltration on Sephadex G-100. They were designated as ${\beta}-1$, 4-mannanase A,B and C, respectively, in the order of isolation. 2) These enzymes were different in several aspects such as pH optimum, effect of metal ions, adsorbability on cellulose derivatives, molecular weight, Michaelis constant toward reduced ivory nut mannan A, mode of action and extent of hydrolysis of the mannan. 3) ${\beta}-1$, 4-Mannanases A and C were proposed to be two different endo-enzymes of random-splitting type producing a series of oligosaccharides from ${\beta}-1$, 4-mannans. ${\beta}-1$, 4-Mannanase B was suggested to be possibly an exe-type enzyme catalyzing a stepwise splitting from the non-reducing end of ${\beta}-1$, 4-mannans to produce mannose. 4) Guaran was subjected to hydrolysis by the purified enzymes and the consequence was discussed in connection with structural requirements of the enzymes toward substituted ${\beta}-1$, 4-mannans and their role in germinating guar seeds.

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Analysis of Bovine Interferon-tau Gene subtypes Expression in the Trophoblast and Non-trophoblast cells

  • Kim, Min-Su;Lim, Hyun-Joo;Lee, Ji Hwan;Park, Soo Bong;Won, Jeong-Il;Kim, Hyun Jong
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.195-203
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    • 2018
  • Interferon-tau (IFNT) is known as a major conceptus protein that signals the process of maternal recognition of pregnancy in ruminants. Also, multiple interferon genes exist in cattle, However, molecular mechanisms of these bovine IFNT (bIFNT) genes whose expressions are limited have not been characterized. We and others have observed that expression levels of bovine subtype IFNT genes in the tissues of ruminants; thus, bIFNT1 and other new type I (bIFNTc1/c2/c3) gene co-exist during the early stages of conceptus development and non-trophoblast cells. Its genes transcription could be regulated through CDX2 and ETS2 and JUN and/or cAMP-response element binding protein (CREB)-binding protein (CREBBP) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. Bovine ear-derived fibroblast cells, were co-transfected with luciferase reporter constructs carrying upstream (positions -1000 to +51) regions of bIFNT1 and other new type I gene and various transcription factor expression plasmids. Compared to each - 1kb-bIFNT1/c1/c2/c3-Luc increased when this constructs were co-transfected with CDX2, ETS2, JUN and/or CREBBP. Also, Its genes was had very effect on activity by CDX2, either alone or with the other transcription factors, markedly increased luciferase activity. However, the degree of transcriptional activation of the bIFNTc1 gene was not similar to that bIFNT1/c2/c3 gene by expression plasmid. Furthermore, Sequence analyses also revealed that the expression levels of bIFNT1/c2/c3 gene mRNAs expression were highest on day 17, 20 and 22 trophoblast and, Madin-Darby bovine kidney (MDBK), Bovine ear-derived fibroblast (EF), and endometrium (Endo) non-trophoblast cells. But, bIFNTc1 mRNA had not same expression level, bIFNTc1 lowest levels than those of IFNT1/c2/c3 gene in both trophoblast and non-trophoblast cells. These results demonstrate that bovine subtype bIFNT genes display differential, in the trophoblast and non-trophoblast cells.

Surgical Treatment for Descending Necrotizing Mediastinitis (하행성 괴사성 종격동염에 대한 수술)

  • Ryu, Kyoung-Min;Seo, Pil-Won;Park, Seong-Sik;Kim, Seok-Kon;Lee, Jae-Woong;Ryu, Jae-Wook
    • Journal of Chest Surgery
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    • v.41 no.1
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    • pp.82-88
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    • 2008
  • Background: Descending necrotizing mediastinitis (DNM) is a serious disease originating in odontogenic or oropharyngeal infection with high mortality despite adequate antibiotics and aggressive surgery. We analyzed results of treatment for DNM. Material and Method: We studied 8 cases diagnosed as DNM from 1998 to 2007. All patients received emergent surgical drainage and debridement with broad spectrum antibiotics just after diagnosis. Antibiotics were changed after bacterial susceptibility testing. The surgical approach included 2 cases of cervicotomy, 6 cases of cervicotomy, and a thoracotomy. Result: The interval between symptom onset and hospitalization was $4.6{\pm}1.8$ days ($1{\sim}9$ day). DNM originated in 4 cases of odontogenic infection (50%), 2 cases of oropharyngeal infection (25%), and 2 cases of unknown origin (25%). Causative organisms were found in 6 cases; Streptococcus in 4 cases, Staphylococcus in 1 case, and Klebsiella in 1 case. The Endo DNM classification was type I (2 cases), IIA (3 cases), and IIB (3 cases). The incidence of thoracotomy was 75%. The surgical mortality rate was 25% (2/8). The cause of death was multiple organ failure caused by septic shock. All mortality cases received only cervicotomy and aggravated infections after initial drainage. Conclusion: Early diagnosis, immediate surgical drainage, and adequate antibiotics, including covered anaerobes, are required. Thoracotomy should be performed with cervicotomy even for localized DNM.

Ability for Biosynthesis of C27 Brassinosteroids by an Enzyme Pool Prepared from Cultured Cells of Phaseolus vulgaris (강낭콩 현탁배양세포에서 추출된 Enzyme Pool의 C27 Brassinosteroids 합성 능력)

  • Kim Tae-Wuk;Joo Se-Hwan;Kim Seong-Ki
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.63-71
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    • 2005
  • A cell-free enzyme solution prepared from suspension cultured cells of Phaseolus vulgaris successfully mediated conversions of cholesterol $\to$ cholestanol and 6-deoxo-28-norteasterone $\leftrightarrow$ 6-deoxo-28-nor-3-dehydroteasterone $\leftrightarrow$ 6-deoxo-28-nortyphasterol $\to$ 6-deoxo-28-norcastasterone $\to$ 28-norcastasterone. Al-though conversion of cholestanol to 6-deoxo-28-norteasterone intermediated by 6-deoxo-28-norcathasterone was not demonstrated, this strongly suggests that a complete set of biosynthetic enzymes catalyzing reactions from cholesterol to 28-norcastasterone via 6-deoxo-28-nor type brassinosteroids is endogenously present in the cells, which demonstrates that a $C_{27}$ brassinosteroids biosynthetic pathway, namely the late C-6 oxidation for $C_{27}$ brassinosteroids, is operative in the cells. Additionally, the enzyme solution mediated conversion of 28-norcastasterone to castasterone in the presence of S-adenosyl-methionine and NADPH, providing that the $C_{27}$ brassinosteroids biosynthesis is an important route to generate castasterone in the cells. Together with our previous finding that castasterone can be biosynthesized by the same biosynthetic pathway in tomato, this study demonstrates that the $C_{27}$ brassinosteroids biosynthesis is a common alternative process to maintain endo-genous level of castasterone, an active $C_{28}$ brassinosteroid, in plants.

Characterization of Endochitosanases-Producing Bacillus cereus P16

  • Jo, Yu-Young;Jo, Kyu-Jong;Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Kil-Yong;Kim, Tae-Hwan;Park, Ro-Dong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.960-968
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    • 2003
  • A bacterial isolate showing a strong endochitosanase activity was isolated from soil and then characterized. The isolate was identified and designated as Bacillus cereus P16, based on morphological and biochemical properties, assimilation tests, cellular fatty acids pattern, along with 16S rRNA gene sequence. The optimized medium for producing extracellular chitosanase in a batch culture contained 1% tryptone, 0.5% chitosan, and 1% NaCl (pH 7.0). Powder chitosan and tryptone served the best as carbon and nitrogen sources, respectively, for the chitosanase production. Chitosanase activity was the highest when culture was completed at $37^{\circ}C$ among various temperatures ($20-42^{\circ}C$) tested in a shaking incubator (200 rpm). The levels of chitosanase activity in the culture fluid were 2.0 U/ml and 3.8 U/ml, respectively, when incubated in a flask for 60 h and in a jar fermenter for 24 h. The culture supernatant showed a strong liquefying activity on the soluble chitosan. The viscosity of 1% chitosan solution, that was incubated with the culture supernatant, was rapidly decreased, suggesting the secretion of endochitosanolytic enzymes by P16. The culture fluid revealed six endo-type chitosanase isozymes, two major (38 and 45 kD), and four minor (54, 65, 82, and 96 kD) forms by staining profile. The crude enzymes were very stable, and full activity was maintained for 4 weeks at $4^{\circ}C\;or\;-20^{\circ}C$ in the culture supernatant, suggesting a highly desirable stability rate for making an industrial application of the crude enzymes. The supernatant also cleaved the insoluble chitosan powder, but the hydrolysis rate was much lower. The enzymic degradation products of chitosan contained $(GlcN)_n$ (n=2-8). The concentration of chitosan in the reaction mixture of the crude enzyme affected the chitooligosaccharides composition of the hydrolysis products. When the higher concentration of chitosan was used, the higher degree of polymerized chitooligosaccharides were produced. By comparison with other commercial chitosanase preparations, P16 was indeed found to be a valuable enzyme source for industrial production of chitooligosaccharides from chitosan.

Effects of electrical stimulation on healing of endo-osseous titanium implants in circumferential defect (전기자극이 성견 골결손부에 매식된 임플란트 주위조직의 치유에 미치는 영향)

  • Shim, Jae-Chang;Kim, Young-Jun;Chung, Hyun-Ju;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.177-193
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    • 2004
  • Several experimental studies showed that the application of small amounts of electric current to bone stimulated osteogenesis at the site of the cathode and suggested that electrical currents promote osseointegration around dental implants. The purpose of this study was to determine the effect of direct microcurrent to endosseous titanium implants placed in bone defects. The right and left 2nd, 3rd and 4th mandibular premolars in ten mongrel dogs (15Kg of weight) were extracted. One monthe later, Ti-machined screw type implants(3.8 mm diameter x 8.5 mm length, $AVANA^{(R)}$, Ostem) were placed in surgically created circumferential defect area(width 5mm, depth 4mm). The implants were divided into three groups according to the treatment modalities: Control group- implants without electrical stimulation; Experimental group I- implants with allogenic demineralized freeze dried bone grafting; and Experimental group II-implants allogenic demineralized freeze dried bone grafting and electric stimulation. The animals were sacrificed in the 4th and 8th week after implant placement and un-decalcified specimens were prepared for histological and histometrical evaluation of bone-implant contact ratio (BIC) and bone formation area ratio (BFA) in defect area. Some specimens at 8 weeks after implantation were used for removal torque testing. Histologically, there was connective tissue infiltration in the coronal part of defect area in control and the experimental group I, whereas direct bone contact was found in the experimental group II without connective tissue invasion. Average BIC ratios at 4 weeks of healing were 60.1% in the experimental group II, 47.4% in the experimental group I and 42.7% in the control. Average BIC ratios at 8 weeks after implantation were 67.6% in the experimental group II, 55.9% in the experimental group I and 54.6% in the control. The average BFA ratio was 84.0% in the experimental group II, 71.8% in the experimental group I and 58.8% in the control at 4 weeks, and the BFA ratios were 89.6% in the experimental group II, 81.4% in the experimental group I and 70.5% in the control at 8 weeks after implantation. The experimental group II showed also significantly greater BIC and BFA ratios compared to the control and the experimental group I (p<0.05). The removal torque values at 8 weeks after implantation were 56 Ncm in the experimental group II, 49 Ncm in the experimental group I and 43 Ncm in the control. There was a statistically significant difference among 3 groups (p<0.05). These results suggest that electrical stimulation improve and accelerate bone healing around endosseous titanium implants in bone defect.

Homology Modeling and Characterization of Oligoalginate Lyase from the Alginolytic Marine Bacterium Sphingomonas sp. Strain MJ-3 (알긴산을 분해하는 해양미생물인 Sphingomonas sp. MJ-3 균주의 올리고알긴산 분해효소의 상동성 모델링 및 특성연구)

  • Kim, Hee Sook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.121-129
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    • 2015
  • Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

Probe of Unfilled Sheath of Prestressed-Concrete Girder Bridge Using Impact-Echo Method (충격탄성파법을 이용한 PC형교의 쉬이스 내부 그라우트 미충전부 탐사)

  • Lee, Sang Hun;Kim, Sang Jin;Endo, Takao;Sagara, Yuzo
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.15 no.1
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    • pp.112-119
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    • 2011
  • Sheaths are arranged in the web and lower part of prestressed-concrete girder bridges, and prestressing wires and concrete are indirectly bonded together by filling interior parts of the sheaths. However, when the filling is not sufficient, water can penetrate the interior parts of the sheaths and, thereby, prestressing wires can be corroded or transverse cracks would occur around sheaths by the freezing and thawing action of the penetrated water. Therefore, it is an important element in maintenance management of structures to find out the sheaths unfilled with grout early after construction. In this paper, in order not to damage bridge members, the impact-echo method with a new approach in application is used to probe sheaths unfilled with grout for real structures. The location of sheaths is first estimated with reinforcing bar probing instrument of radar type and the measurement locations of sheaths are determined. By sending elastic wave to the side of girders and receiving the response, the location of the unfilled part of a sheath was estimated from the difference between high frequency peak and twice the resonance frequency indicating thickness. To verify the location of void estimated by the impact-echo method, pictures were taken by an industrial fiber scope after drilling a hole.

Study on the Availability of Repeated Flexible Bronchoscopy(RFB) (반복적 굴곡성 기관지경검사(RFB)의 유용성에 대한 연구)

  • Lee, Hong-Lyeol;Moon, Tae-Hoon;Cho, Jae-Hwa;Ryu, Jeong-Seon;Kwak, Seung-Min;Cho, Chul-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.365-376
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    • 2000
  • Background : Ever since Flexible Fiberoptic Bronchoscopy was introduced into clinical practice, it has played an important role in both diagnosis and therapy of respiratory diseases. Repeated bronchoscopic examinations is are not so uncommon. This study was designed prospectively to assess the clinical availability of the Repeated Flexible Bronchoscopy (RFB). Methods : Pre-established indications were as follows : 1) To confirm diagnosis or the cell type in proven malignancy 2) to diagnose or locate hemoptysis 3) to follow-up or confirm recurrence 4) to use in therapy. We performed RFB and analyzed the data in 156 patients during 28-month period. Results : The frequency of RFB was 23.0%. The indication for diagnosis or cell type of malignancy was 25 cases, of which 2 cases were confirmed by a third bronchoscopic examination and 3 cases by surgical procedures. Localization of the bleeding site was confirmed in 53.8%. RFB for small cell lung cancer yielded more information on residual or recurred lesion not apparent even with the CT scan in 30%. Previous cases of bronchostenosis due to endo-bronchial tuberculosis was shown to have worsened in 66.7%. Therapeutic manipulations were done in 126 cases, and bronchial suction was most common. Complications showed decreasing tendency with repeated examinations. Conclusion : The RFB for diagnosis or cell type of malignancy was useful in that comfirmation of diagnosis was possible in 85.7% of malignancy. More aggressive procedures should be employed including TBLB or TBNA. The RFB showed possible usefulness in the follow-up of patients with small cell lung cancer. For the patients with hemoptysis or endobronchial tuberculosis, the RFB did not the significance did not show significance because its results did not influence the diagnosis, therapy or clinical course.

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