• Mass Spectrometry Letters
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• v.12 no.3
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• pp.66-75
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• 2021

Interleukin-4 (IL-4) and IL-13 are cytokines secreted by immune cells. Cytokines induce the proliferation of macrophages or promote the differentiation of secretory cells. The initiation and progression of allergic inflammatory diseases, such as asthma, are dependent on cytokines acting through related receptor complexes. IL-4 and IL-13 are N-glycoproteins. Glycan structures in glycoproteins play important roles in protein folding, protein stability, enzymatic function, inflammation, and cancer development. Therefore, the glycan structure of IL-4 and IL-13 needs to be elucidated in detail for the development of effective therapies. We report the first attempt to characterize the site-specific N-glycosylation of recombinant IL-4 and IL-13 via liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The tandem mass spectra of intact N-glycopeptides were identified using the Integrated GlycoProteome Analyzer (I-GPA) platform, which can automatically and rapidly analyze multiple N-glycopeptides, including their glycan composition and amino acid sequences. The recombinant IL-4 and IL-13 were identified with amino acid sequence coverages of 84% and 96%, respectively. For IL-4, 52 glycoforms on one N-glycosylation site were identified and quantified. In IL-13, 232 N-glycopeptides from three N-glycosylation sites were characterized, with the site Asn52 being the most extensively glycosylated (~80%). The complex glycans were the most abundant glycan on IL-4 and IL-13 (~96% and 91%, respectively), and the biantennary glycans were the most abundant in both recombinant IL-4 and IL-13 proteins.

• Journal of Korean Clinical Nursing Research
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• v.28 no.3
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• pp.277-288
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• 2022

Purpose: This study was conducted to evaluate the effects of the insulin therapy adherence program (INSTA GRAM) for patients with type 2 diabetes mellitus. Methods: This study examined a total of 63 patients with type 2 diabetes for whom the initiation of insulin therapy is being considered or within 6 months of undergoing insulin therapy. INSTAGRAM is a stage-specific intervention consisting of strategies for motivating patients to initiate and maintain insulin therapy using the transtheoretical model on the stages of change, the process of change, and self-efficacy. The participants were randomly assigned to either the INSTAGRAM group (experimental group, n=32) or the standard diabetes education group (control group, n=31). The INSTAGRAM group received 6 sessions over 8 weeks(face-to-face education was conducted in the first and last sessions, and telephone coaching was conducted from the second to fifth sessions). The outcomes were measured by psychological insulin resistance, self-efficacy, and glycosylated hemoglobin (HbA1c). Results: Psychological insulin resistance (p<.001), self-efficacy (p<.001), and HbA1c (p=.024) of the participants in the INSTAGRAM group significantly improved compared to those in the control group. Conclusion: INSTAGRAM is effective in overcoming psychological insulin resistance and improving self-efficacy and HbA1c. Therefore, the INSTAGRAM can be recommended as a nursing intervention for type 2 diabetic patients who delay the initiation of insulin therapy.

• Journal of Veterinary Science
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• v.21 no.1
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• pp.5.1-5.12
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• 2020

The major glycoproteins of bovine gammaherpesvirus 4 (BoHV-4) are gB, gH, gM, gL, and gp180 with gB, gH, and gp180 being the most glycosylated. These glycoproteins participate in cell binding while some act as neutralization targets. Glycosylation of these envelope proteins may be involved in virion protection against neutralization by antibodies. In infected cattle, BoHV-4 induces an immune response characterized by low neutralizing antibody levels or an absence of such antibodies. Therefore, virus seroneutralization in vitro cannot always be easily demonstrated. The aim of this study was to evaluate the neutralizing capacity of 2 Argentine BoHV-4 strains and to associate those findings with the gene expression profiles of the major envelope glycoproteins. Expression of genes coding for the envelope glycoproteins occurred earlier in cells infected with isolate 10/154 than in cells infected with strain 07/435, demonstrating a distinct difference between the strains. Differences in serological response can be attributed to differences in the expression of antigenic proteins or to post-translational modifications that mask neutralizing epitopes. Strain 07/435 induced significantly high titers of neutralizing antibodies in several animal species in addition to bovines. The most relevant serological differences were observed in adult animals. This is the first comprehensive analysis of the expression kinetics of genes coding for BoHV-4 glycoproteins in 2 Argentine strains (genotypes 1 and 2). The results further elucidate the BoHV-4 life cycle and may also help determine the genetic variability of the strains circulating in Argentina.

• The Journal of Korean Academy of Orthopedic Manual Physical Therapy
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• v.29 no.3
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• pp.63-72
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• 2023

Background: This study investigated the effects of face-to-face and non-face-to-face McKenzie exercises and elastic band strengthening exercises on falls and fitness in older adults. Methods: A total of 44 subjects were divided into a face-to-face group (FG; n=22) and a non-face-to-face group (NFG; n=22) who met older adults. Both groups used McKenzie exercises and strength training with elastic bands The fall efficacy scale (K-FES) and glycosylated hemoglobin (HbA1c) Pro were used to compare the senior fitness test (SFT). Results: No significant differences were detected between the two groups before the intervention; however, a significant difference was noted in the K-FES scores after the intervention. The change in HbA1c after the intervention was not statistically significant in the FG or the NFG, and no significant difference was detected between the two groups. The change in lower limb muscle strength in the elderly physical fitness test showed no significant difference between the two groups before or after the intervention. The change in upper limb flexibility after the intervention was not significantly different in the FG or NFG, indicating no significant difference between the two groups. The change in the 3-meter round-trip test after the intervention was significantly different in both the FG and NFG groups, but the difference between the groups was not statistically significant. The change in the grip strength test of the dominant hand showed no significant difference after the intervention in the FG or the NFG, and the difference between the FG and NFG groups was not statistically significant. The dominant foot external standing after the intervention was significantly improved in the FG, but no significant difference was detected in the NFG, and the difference between the two groups was not statistically significant. Conclusion: The study findings revealed that FG and NFG McKenzie exercises and elastic band strengthen exercises were equally effective at preventing falls and improving fitness in older adults.

• Journal of Microbiology and Biotechnology
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• v.34 no.4
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• pp.930-939
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• 2024

Mushroom laccases play a crucial role in lignin depolymerization, one of the most critical challenges in lignin utilization. Importantly, laccases can utilize a wide range of substrates, such as toxicants and antibiotics. This study isolated a novel laccase, named HeLac4c, from endophytic white-rot fungi Hericium erinaceus mushrooms. The cDNAs for this enzyme were 1569 bp in length and encoded a protein of 523 amino acids, including a 20 amino-acid signal peptide. Active extracellular production of glycosylated laccases from Saccharomyces cerevisiae was successfully achieved by selecting an optimal translational fusion partner. We observed that 5 and 10 mM Ca²⁺, Zn²⁺, and K⁺ increased laccase activity, whereas 5 mM Fe²⁺ and Al³⁺ inhibited laccase activity. The laccase activity was inhibited by the addition of low concentrations of sodium azide and ⳑ-cysteine. The optimal pH for the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt was 4.4. Guaiacylglycerol-β-guaiacyl ether, a lignin model compound, was polymerized by the HeLac4c enzyme. These results indicated that HeLac4c is a novel oxidase biocatalyst for the bioconversion of lignin into value-added products for environmental biotechnological applications.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.8
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• pp.1086-1093
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• 2012

The hypoglycemic and hypolipidemic effects of autoclaved soy flour and DJI chungkukjang powder fermented using Bacillus subtilis DJI were investigated in type 2 diabetic animal models. After a 2-week adaptation period, the diabetic animal model db/db mice were divided into the diabetic control group (D-C group), a diabetic group fed with soybean (D-S group), and a diabetic group fed with DJI chungkukjang (D-CJ group). The body weight gain, food intake, water intake, liver, and adipose tissue weights were not significantly different between the experimental groups. The supplementation of DJI chungkukjang or autoclaved soy flour diet induced a marked reduction of fasting blood glucose, blood glycosylated hemoglobin levels, and glucose levels in the oral glucose tolerance test and AUC for glucose compared with the diabetic control group. However, DJI chungkukjang showed a much stronger antidiabetic effect than unfermented autoclaved soy flour. Serum insulin levels were the same among the groups. The supplementation of DJI chungkukjang or autoclaved soy flour diet also significantly lowered the serum triglyceride, total cholesterol, and LDL-cholesterol levels compared with the control diabetic group, while it elevated the HDL-cholesterol level in the serum. This data suggests that the dietary supplementation of autoclaved soy flour or DJI chungkukjang may be useful in the control of blood glucose in animals with type 2 diabetes.

• Journal of Life Science
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• v.20 no.4
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• pp.584-588
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• 2010

Clusterin is an 80 kDa heterodimetric glycosylated protein which plays diverse biological roles in various tissues and organs. Clusterin is reported to be associated with the pathogenesis of coronary artery disease and atherosclerosis. Therefore, we investigated the genotype for the T

• Journal of Life Science
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• v.22 no.2
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• pp.142-147
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• 2012

The sporulation-specific glucoamylase (SGA) of Saccharomyces diastaticus is known to be produced in the cytoplasm during sporulation. For the purpose of proving that SGA has secretory potential, we constructed a hybrid plasmid, pYESC25, containing the promoter and the putative signal sequence of the SGA fused in frame to the endo-1,4-${\beta}$-D-glucanase (CMCase) gene of Bacillus subtilis without its own signal sequence. The recipient yeast strain of S. diastaticus YIY345 was transformed with the hybrid plasmid. CMCase secretion from S. diastaticus harboring pYESC25 into culture medium was confirmed by the formation of yellowish halos around transformants after staining with Congo red on a CMC agar plate. The transformant culture was fractionated to the extracellular, periplasmic, and intracellular fraction, followed by the measurement of CMCase activity. About 63% and 13% enzyme activity were detected in the culture supernatant (extracellular fraction) and periplasmic fraction, respectively. Furthermore, ConA-Sepharose chromatography, native gel electrophoresis, and activity staining revealed that CMCase produced in yeast was glycosylated and its molecular weight was larger than that of the unglycosylated form from B. subtilis. Taking these findings together, SGA has the potential of secretion to culture medium, and the putative signal sequence of SGA can efficiently direct bacterial CMCase to the yeast secretion pathway.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.195-200
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• 2008

Saxifraga stolonifera (Saxifragaceae) is a perennial herbaceous plant growing in Korea, China, Japan and Russia. The aerial part has been used as herbal medicine for treatment of pneumonia, frostbite, inflammation and microbial infection. In this study, fresh juice and methanol extract were prepared from the aerial part of S. stolonifera, and their antimicrobial, antithrombin, and antioxidant activity were evaluated, respectively. The fresh juice showed weak antimicrobial activity against Proteus vulgaris, Escherichia coli O157:H7 and Candida albicans with ignorable DPPH scavenging activity. But, the methanol extract showed strong antioxidant activity ($IC_{50}$ of $37.5{\mu}g/mL$) with minor, broad-range antimicrobial activity. Antithrombin activities were not observed in fresh juice and extract, up to 1.5 mg/mL. Sequential organic solvent fractionation of methanol extract showed that $IC_{50}s$ of ethylacetate and the butanol fraction were 6.9 and $7.8{\mu}g/mL$, respectively, that is comparable with vitamin C or butylated hydroxytoluene. Analysis of component in extract and fractionates suggested that the antioxidants in fractions are diverse and the active substances have glycosylated phenolic structure. Our results suggest that the aerial part of S. stolonifera could be used as the natural source of potential antioxidant.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.35-35
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• 2005

Recombinant human $interferon-{\beta}-1a(rIFN-{\beta})$ is a single glycosylated protein (at N80, 1N) with anti-viral activity. However, present drugs have a relatively short serum half-life of $rIFN-{\beta}$, thus patients suffer from frequent $infections.^{1)}$ To improve its half-life, eight glycosylation analogs were prepared, which have additional N-linked glycosylation consensus sequences (N-X-T/S) within the $IFN-{\beta}$ molecule and/or at C-terminal. Each $rIFN-{\beta}$ analog was examined for the presence of additional N-linked glycosylation and the maintenance of anti-viral activity in CHO cells. The molecular weights of five analogs were not changed. However, two analogs, R27T within $rIFN-{\beta}$ (27 kDa, 2N) and GNITVNITV at C-terminal (29kDa, 2N), showed a clear increase in molecular weights, compared to native $rIFN-{\beta}$ (23 kDa, 1N). And another combined analog of R27T+GNITVNITV showed increased molecular weight (33 kDa, 3N). It was confimed that the molecular weight increment of analogs was caued by the N-linked glycosylation with the treatment of N-glycansae. In the case of anti-viral activity, the analog GNITVNITV showed a reduction in activity compared to native $IFN-{\beta}$, whereas the analogs R27T and R27T+GNITVNITV were found to have distinctly increased activities. Pharmacokinetic study in rats also disclosed that the analogs R27T and R27T+GNITVNITV had 2 3 fold increased serum half-life, respectively. In conclusion, the addition of N-linked glycosylation in $rIFN-{\beta}$ increased serum half-life, thereby its less frequent administration will be expected.