• Title/Summary/Keyword: Sialic acids

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Detection of Viruses and Changes of Protein of Saliva in Patients with Recurrent Aphthous Ulcer (구내 재발성 아프타성 궤양 환자에서 타액내 바이러스 검출 및 단백질의 변화)

  • Park, Sang-Bae;Kim, Byung-Gook;Bae, Jeong-Sik
    • Journal of Oral Medicine and Pain
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    • v.24 no.2
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    • pp.125-135
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    • 1999
  • This study was performed to discover the underlining influences of Herpes Simplex virus (HSV) and Varicella Zoster virus (VZV), to detect the changes of whole protein and mucin level and to observe protein profiles in the saliva when recurrent aphthous ulcer (RAU) was present. Unstimulated whole saliva was collected from 23 patients who for over three years had a clinical history of RAU, in a group of 10 women and 13 men, ranging from 11 to 72 years of age, and 20 healthy subjects, in a group of 8 women and 12 men, who did not have the symptoms nor a past history of RAU. Through the means of Polymerization Chain Reaction, genomic DNA from the HSV and VZV was purified from the saliva samples for identifying precisely the two types of viruses, and the level of whole protein and sialic acids in the saliva and the ratio of sialic acid to whole protein were measured, and SDS-polyacrylamide gel electrophoresis was performed. The results obtained were as follows ; 1. 39.13% of patients showed 224 bp bands of VZV DNA, those were appeared more in patients than in control group (p<0.01), but there was no significant difference between patients and control group in HSV DNA (p>0.05). 2. The concentration of whole protein in men patients was lower than in men control group (p<0.05), but there were no significant differences between patients and control in other groups (p>0.05). 3. The concentration of sialic acids from patients was lower than control group in all groups (p<0.05). 4. The concentration of sialic acids in proportion to that of whole protein was lower in patients than in control group (p<0.05), and in the two women groups (p<0.01), but no noticeable difference was found between the two men groups (p>0.05). 5. There were no consistent differences observed in the protein profiles of patients with control group except that certain protein bands near 50 kDa was lower in patients than in control group. These results suggest that viruses such as HSV and VZV and reduction of salivary whole protein and mucin levels are related to development of RAU.

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Enhanced sialylation and in vivo efficacy of recombinant human α-galactosidase through in vitro glycosylation

  • Sohn, Youngsoo;Lee, Jung Mi;Park, Heung-Rok;Jung, Sung-Chul;Park, Tai Hyun;Oh, Doo-Byoung
    • BMB Reports
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    • v.46 no.3
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    • pp.157-162
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    • 2013
  • Human ${\alpha}$-galactosidase A (GLA) has been used in enzyme replacement therapy for patients with Fabry disease. We expressed recombinant GLA from Chinese hamster ovary cells with very high productivity. When compared to an approved GLA (agalsidase beta), its size and charge were found to be smaller and more neutral. These differences resulted from the lack of terminal sialic acids playing essential roles in the serum half-life and proper tissue targeting. Because a simple sialylation reaction was not enough to increase the sialic acid content, a combined reaction using galactosyltransferase, sialyltransferase, and their sugar substrates at the same time was developed and optimized to reduce the incubation time. The product generated by this reaction had nearly the same size, isoelectric points, and sialic acid content as agalsidase beta. Furthermore, it had better in vivo efficacy to degrade the accumulated globotriaosylceramide in target organs of Fabry mice compared to an unmodified version.

Enhanced γ-aminobutyric acid and sialic acid in fermented deer antler velvet and immune promoting effects

  • Yoo, Jiseon;Lee, Juyeon;Zhang, Ming;Mun, Daye;Kang, Minkyoung;Yun, Bohyun;Kim, Yong-An;Kim, Sooah;Oh, Sangnam
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.166-182
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    • 2022
  • Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines

  • Talabnin, Krajang;Talabnin, Chutima;Ishihara, Mayumi;Azadi, Parastoo;Wongkham, Sopit;Sripa, Banchob
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.691-695
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    • 2016
  • Protein glycosylation is the most common posttranslational modification in mammalian cells. Aberrant protein glycosylation has been reported in various diseases, including cancer. We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-$MS^n$). Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by $NeuA_{c2}Gal_1GalNA_{c1}$. These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA.

Isolation of New CHO Cell Mutants Defective in CMP-Sialic Acid Biosynthesis and Transport

  • Shin, Dong-Jun;Kang, Ji Young;Kim, Youn Uck;Yoon, Joong Sik;Choy, Hyon E;Maeda, Yusuke;Kinoshita, Taroh;Hong, Yeongjin
    • Molecules and Cells
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    • v.22 no.3
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    • pp.343-352
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    • 2006
  • Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

Anti-bacterial effects of enzymatically-isolated sialic acid from glycomacropeptide in a Helicobacter pylori-infected murine model

  • Noh, Hye-Ji;Koh, Hong Bum;Kim, Hee-Kyoung;Cho, Hyang Hyun;Lee, Jeongmin
    • Nutrition Research and Practice
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    • v.11 no.1
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    • pp.11-16
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    • 2017
  • BACKGROUND/OBJECTIVES: Helicobacter pylori (H. pylori) colonization of the stomach mucosa and duodenum is the major cause of acute and chronic gastroduodenal pathology in humans. Efforts to find effective anti-bacterial strategies against H. pylori for the non-antibiotic control of H. pylori infection are urgently required. In this study, we used whey to prepare glycomacropeptide (GMP), from which sialic acid (G-SA) was enzymatically isolated. We investigated the anti-bacterial effects of G-SA against H. pylori in vitro and in an H. pylori-infected murine model. MATERIALS/METHODS: The anti-bacterial activity of G-SA was measured in vitro using the macrodilution method, and interleukin-8 (IL-8) production was measured in H. pylori and AGS cell co-cultures by ELISA. For in vivo study, G-SA 5 g/kg body weight (bw)/day and H. pylori were administered to mice three times over one week. After one week, G-SA 5 g/kg bw/day alone was administered every day for one week. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), IL-$1{\beta}$, IL-6, and IL-10 levels were measured by ELISA to determine the anti-inflammatory effects of G-SA. In addition, real-time PCR was performed to measure the genetic expression of cytotoxin-associated gene A (cagA). RESULTS: G-SA inhibited the growth of H. pylori and suppressed IL-8 production in H. pylori and in AGS cell co-cultures in vitro. In the in vivo assay, administration of G-SA reduced levels of IL-$1{\beta}$ and IL-6 pro-inflammatory cytokines whereas IL-10 level increased. Also, G-SA suppressed the expression of cagA in the stomach of H. pylori-infected mice. CONCLUSION: G-SA possesses anti-H. pylori activity as well as an anti-H. pylori-induced gastric inflammatory effect in an experimental H. pylori-infected murine model. G-SA has potential as an alternative to antibiotics for the prevention of H. pylori infection and H. pylori-induced gastric disease prevention.

Production and Characterization of a Monoclonal Antibody against Surface Glycoprotein, gp6 1, on K562 Erythroleukemia Cells (K562 적혈구암 세포주의 표면 당단백질에 대한 단클론항체의 생성 및 특성)

  • 김한도;정재훈;홍선화;김정락;한규형;임운기;유미애;이경희;강호성
    • The Korean Journal of Zoology
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    • v.39 no.1
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    • pp.12-20
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    • 1996
  • A multipotential hematopojetic cell line, 1(562 cell, was differentiated into megakaryocyte by a chemical inducer, PMA, with an enhanced expression of gpIlla accompaning with a distinct morphological change. On the other hand, 1(562 cells were differentiated into erythrocytes by other chemical inducers, DMSO or butyrate, with a concomitant increase in hemoglobin accumulation. An antigen of apparent molecular weight of 61 kDa was identified on the surface of 1(562 cells by using monoclonal antibody raised against 1(562 cells. The antigen was considered to be a glycoprotein molecule rich in sialic acids and the epitope of antigen was sensitive to neuraminidase digestion or peroxidase oxidation, but resistant to heat treatment. The 61 kDa surface antigen was increased or decreased in its expression along differentiation of 1(562 cells into megakaryocytes or erythrocytes, respedively.

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진흙버섯의 항인플루엔자 활성 및 활성성분 규명

  • Hwang, Byung Soon;Yun, Bong-Sik
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.41-41
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    • 2016
  • Influenza viruses are RNA viruses that belong to the Orthomyxoviridae family, and those can be divided into three types; A, B, and C, which based on the differences of the inner nucleoproteins and genomic structures. All three genera differ in their genomic structure and nucleoprotein content, they are further classified into various serotypes based on the two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). These glycoproteins play crucial roles in viral infection and replication. Hemagglutinin mediates binding of virions to sialic acid receptors on the surfaces of target cells at the initial stage of infection. Neuraminidase cleaves the glycosidic bonds of sialic acids from the viral and cell surfaces to release the mature virions from infected cells, after viral replication. Because NA plays an important role in the viral life cycle, it is considered an attractive therapeutic target for the treatment of influenza. The methanolic extracts of Phellinus baumii and Phellinus igniarius exhibited significant activity in the neuraminidase inhibition assay. Polyphenolic compounds were isolated from the methanolic extracts. The structures of these compounds were determined to be hispidin, hypholomine B, inoscavin A, davallialactone, phelligridin D, phelligridin E, and phelligridin G by spectroscopic methods. Compounds inhibited the H1N1 neuraminidase activity in a dose-dependent manner with $IC_{50}$ values of 50.9, 22.9, 20.0, 14.2, 8.8, 8.1 and $8.0{\mu}M$, respectively. Moreover, these compounds showed anti-influenza activity in the viral cytopathic effect (CPE) reduction assay using MDCK cells. These results suggests that the polyphenols from P. baumii and P. igniarius are promising candidates for prevention and therapeutic strategies against viral infection.

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