• Journal of the Korean Chemical Society
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• v.42 no.5
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• pp.549-558
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• 1998

O-Antigenic part of Campylobacter jejuni gram negative bacteria was reported to consist of a repeated trisaccharide unit. The disaccharides, GlcNAc-Gal derivatives, as key intermediates for the synthesis of trisaccharide repeating units were synthesized. At the $\beta(1{\to}3)$ glycoside bond formation step between 3,4,6-tri-O-acctyl-2-deoxy-2-N-phthalimido-${\beta}$-D-glucopyranosyl bromide and galactosyl acceptors, high regioselectivities between 2, 3, and 4-OH groups of galactosyl acceptors were found. As a result, no further selective protection steps for OH groups of galactosyl acceptors was necessary, and more effective and compact synthetic scheme was achieved.

• Journal of Microbiology and Biotechnology
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• v.20 no.1
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• pp.193-201
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• 2010

A specific and rapid real-time PCR assay for detecting Ralstonia solanacearum in horticultural soil and plant tissues was developed in this study. The specific primers RSF/RSR were designed based on the upstream region of the UDP-3-O-acyl-GlcNAc deacetylase gene from R. solanacearum, and a PCR product of 159 bp was amplified specifically from 28 strains of R. solanacearum, which represent all genetically diverse AluI types and all 6 biovars, but not from any other nontarget species. The detection limit of $10^2\;CFU/g$ tomato stem and horticultural soil was achieved in this real-time PCR assay. The high sensitivity and specificity observed with field samples as well as with artificially infected samples suggested that this method might be a useful tool for detection and quantification of R. solanacearum in precise forecast and diagnosis.

• Journal of Microbiology and Biotechnology
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• v.19 no.7
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• pp.727-733
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• 2009

Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.289-295
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• 2013

Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of ${\alpha}1$,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human $EF1{\alpha}$ promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human $EF1{\alpha}$ promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.274-280
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• 2005

A chitinase was purified from the culture supernatant of Paenibacillus illinoisensis KJA-424 by protein precipitation, DEAE-Sephadex anion-exchange chromatography, and Sephadex G-150 gel filtration. The molecular weight of the purified chitinase was 54 kDa on SDS-PAGE and activity staining. Optimal pH and temperature were pH 5.0 and 60$^{circ}$C, the presence of 10 ruM Ag$^{+}$ and Hg$^{2+}$ inhibited the activity by $92.1/%$ and $97.7/%$, and the K$_{m}$ and V$_{max}$ values were 1.12 mg chitin mrl and 1.48$\mu$mol GlcNAc min$^{-1}$, respectively. The enzyme hydrolyzed tetramer to dimer, pentamer to dimer and trimer, and hexamer to dimer, trimer and tetramer, indicating an endo-splitting mechanism. The chitinase had no hydrolytic activity toward dimer and trimer. The chitinase inhibited the mycelial growth of Rhizoctonia solani, suggesting an antifungal property.

• Experimental and Molecular Medicine
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• v.50 no.12
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• pp.9.1-9.12
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• 2018

Endometriosis is a disease characterized by implants of endometrial tissue outside the uterine cavity and is strongly associated with infertility. Focal adhesion of endometrial tissue to the peritoneum is an indication of incipient endometriosis. In this study, we examined the effect of various cytokines that are known to be involved in the pathology of endometriosis on endometrial cell adhesion. Among the investigated cytokines, transforming growth factor-${\beta}1$ ($TGF-{\beta}1$) increased adhesion of endometrial cells to the mesothelium through induction of ${\alpha}2-6$ sialylation. The expression levels of ${\beta}$-galactoside ${\alpha}2-6$ sialyltransferase (ST6Gal) 1 and ST6Gal2 were increased through activation of $TGF-{\beta}RI/SMAD2/3$ signaling in endometrial cells. In addition, we discovered that terminal sialic acid glycan epitopes of endometrial cells engage with sialic acid-binding immunoglobulin-like lectin-9 expressed on mesothelial cell surfaces. Interestingly, in an in vivo mouse endometriosis model, inhibition of endogenous sialic acid binding by a $NeuAc{\alpha}2-6Gal{\beta}1$-4GlcNAc injection diminished $TGF-{\beta}1$-induced formation of endometriosis lesions. Based on these results, we suggest that increased sialylation of endometrial cells by $TGF-{\beta}1$ promotes the attachment of endometrium to the peritoneum, encouraging endometriosis outbreaks.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.17 no.3
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• pp.85-91
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• 2017

Purpose: The aim of this study was to describe the clinical and biochemical features as well as the molecular analysis of a newly diagnosed illustrative case with ML II and to analyze the clinical features of 11 Korean patients with ML II/III. Method: Including a newly diagnosed patient, total 11 patients in 10 families were diagnosed as ML II (n=7) or ML III (n=4) were enrolled in the study. A diagnosis of ML II or III was made by demonstrating increased lysosomal enzyme activities in the plasma and sequence analysis of GNPTAB with characteristic clinical features. Result: A illustrative case of ML II patient was a 17 month-old boy showing characteristic facial appearance, multiple joint contractures with cardiac involvements. The enzyme assay showed increased lysosomal enzyme activities in the plasma. We identified compound heterozygous mutations in GNPTAB sequence analysis, including a frameshift (c.3428dupA [pAsn1143Lysfs*3]) and a nonsense variant c.673C>T (p.Gln225*). In total 11 patients with ML II/III, the patients with ML II showed severe growth retardation (height standard deviation score -3.2 [${\pm}1.5$]), compare to patients with ML III. Furthermore, patients with ML II patients had serious cardiac problem (n=4), hepatomegaly (n=3) and underwent tracheostomy (n=3) with further respiratory support due to respiratory distress. To improve osteoporosis and bone pain, all patients with ML III and four of 7 patients with ML II treated with intravenous pamidronate. Conclusion: Here we showed a newly diagnosed case of ML II and clinical features of 11 Korean patients with ML II or III. These data could be helpful for further diagnosis of mucolipidosis, a rare inherited metabolic disease, in Korea.

• Korean Journal of Cognitive Science
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• v.13 no.3
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• pp.1-10
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• 2002

This study was performed to analyze the physical attributes of the faces and affective words on the fares. For analyzing physical attributes inside of a face, 36 facial features were selected and almost of them were the lengths or distance values. For analyzing facial contour 14 points were selected and the lengths from nose-end to them were measured. The values of these features except ratio values normalized by facial vortical length or facial horizontal length because the face size of each person is different. The principal component analysis (PCA) was performed and four major factors were extracted: 'facial contour' component, 'vortical length of eye' component, 'facial width' component, 'eyebrow region' component. We supposed the five-dimensional imaginary space of faces using factor scores of PCA, and selected representative faces evenly in this space. On the other hand, the affective words on faces were collected from magazines and through surveys. The factor analysis and multidimensional scaling method were performed and two orthogonal dimensions for the affections on faces were suggested: babyish-mature and sharp-soft.

• Journal of the Korean Chemical Society
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• v.40 no.8
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• pp.557-564
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• 1996

FAB CAD MS/MS(Fast Atom Bombardment Collision Activated Dissociation Mass Spectrometry/Mass Spectrometry) was used to study different degree of bond stability according to the linkage positions of alkali cationized $(Na^+, Li^+, K^+, NH_4^+)$ stereoisomeric and synthetic oligosaccharides. The alkali metal cations were much more stable, requiring over -40 eV of collision energy vs. only -10 eV for the protonated forms. Of the cations, the potassium cationized trisaccharides were more stable than the others. They would not yield fragment ions under the conditions of collision available in triple quadrupole. Other cationized species exhibited decreasing stability in the series $Nap^+>Li^+>NH_4^+$ using 0.8 mTorr argon pressure in the collision cell. Metal cations of the oligosaccharides maintained charge principally on the amino sugar as shown by shift of all the fragment ions containing the amino sugar. The reason for the higher stability of the metal cationized form is the formation of crown ether-like bond around metal cations, N-acetyl group on GlcNAc and oxygens on fucose moiety. Depending on the metal sizes and the conformation of linkage-isomeric region, cationized species gave linkage dependent fragment patterns and exhibited stability in the series 1-6 > 1-4 > 1-3 linkage.

• Korean Journal of Environmental Agriculture
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• v.14 no.3
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• pp.319-328
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• 1995

This study was carried out to investigate the resistance mechanism by three different kinds of procymidone-resistant and susceptible isolates of Botrytis cinerea which had been collected from green houses. The average resistance level of the resistant strains was 1,000 times higher than that of susceptible ones. Also, it was revealed that the resistance was not originated from components excreted by Botrytis cinerea, based on the result obtained from the treatment with piperonyl butoxide and triphenyl phosphate as an inhibitor of monooxygenase and esterase, respectively. The total lipod content of resistant strains was 1.3 times higher than that of susceptible ones, among fatty acids, palmitic acid, stearic acid, and linoleic and being 3.0, 2.5, and 2.0 times higher, respectinely. Also slight differences in sterol contents and components were observed. The crude chitin content was slightly higher in susceptible strains but contents of N-acetyl glucosamine, a hydrolysate of chitin, were about 2 times higher in resistant ones.