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Bing Liu

Publications and source records attributed to Bing Liu.

96 records · Page 6Linked to original sources

Structural characterisation of cysteines in a bacterial-binding motif of human salivary mucin MG2.

Human salivary mucin MG2 is a 180 kDa glycoprotein secreted by submandibular/sublingual and minor salivary glands. Secreted MG2 contains a domain with the only two cysteines (Cys(45) and Cys(50)) present in the polypeptide backbone; in native and recombinant MG2 this domain is involved in mucin binding to oral microbes. As the reduction and alkylation of MG2 has been shown to abolish binding, the present study was undertaken to determine whether the cysteine residues exist in the dithiol or disulphide form. Electrophoretic analysis under reducing and non-reducing conditions showed that intermolecular disulphide bonds do not occur between MG2 molecules. The same incorporation of radiolabelled iodoacetamide into MG2 was obtained with or without prior reduction. When radiolabelled alkylated MG2 was digested with Endoproteinase Lys-C and the derived peptides were separated by reversed-phase high-performance liquid chromatography (RP-HPLC), radioactivity was found in two fractions. Mass spectral analyses of these fractions showed the presence of peptides Cys-Leu-His-Lys and Arg-Cys-Arg-Pro-Lys, both containing carboxymethylated cysteines. These results show that the cysteines in the structural motif associated with bacterial binding exist in the dithiol form, and suggest the potential use of cysteine-containing peptides as agents to modify interactions of MG2 with microbes and oral surfaces.

Amino Acid Motifs↗

Interaction of human salivary mucin MG2, its recombinant N-terminal region and a synthetic peptide with Actinobacillus actinomycetemcomitans.

The antimicrobial properties of human salivary mucin MG2 against the periodontal pathogen, Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans), were investigated using purified MG2, rNMUC7 (a recombinant polypeptide containing residue 1-144 of MG2) and synthetic peptides PEP1 (residue 1-17) and PEP2 (residue 47-63). MG2 and rNMUC7 bound to A. actinomycetemcomitans strains SUNY75, SUNY465, SUNY523, 652 and JP2 in a liquid phase binding assay. The bactericidal activities of rNMUC7, PEP1 and PEP2 against A. actinomycetemcomitans SUNY523 were examined in a colony forming unit killing assay. The LD50 for rNMUC7 was 9 microM, for PEP2 was 20 microM and PEP1 did not exhibit bactericidal activity. The primary structure of these polypeptides was analyzed and a direct relationship between net positive charge and bactericidal activity was found. Screening of saliva samples from 60 individuals on Western blots probed with an anti-MG2 antibody against PEP2 revealed that a 20 kDa MG2 fragment was present in 66% of subjects and that this fragment was not present in glandular secretions. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of tryptic peptides derived from the 20 kDa fragment confirmed that this fragment contained a portion of the amino terminal region of MG2. The present study showed that the N-terminal region of MG2 and a subdomain within this region are microbicidal against A. actinomycetemcomitans and that a 20 kDa fragment of MG2 occurs in whole saliva. This suggests that cleavage of MG2 in vivo may produce fragments with microbicidal properties and that this may represent a novel mechanism of host defense.

Adult↗

Expression of membrane-associated mucins MUC1 and MUC4 in major human salivary glands.

Mucins are high molecular weight glycoproteins secreted by salivary glands and epithelial cells lining the digestive, respiratory, and reproductive tracts. These glycoproteins, encoded in at least 13 distinct human genes, can be subdivided into gel-forming and membrane-associated forms. The gel-forming mucin MUC5B is secreted by mucous acinar cells in major and minor salivary glands, but little is known about the expression pattern of membrane-associated mucins. In this study, RT-PCR and Northern blotting demonstrated the presence of transcripts for MUC1 and MUC4 in both parotid and submandibular glands, and in situ hybridization localized these transcripts to epithelial cells lining striated and excretory ducts and in some serous acinar cells. The same cellular distribution was observed by immunohistochemistry. Soluble forms of both mucins were detected in parotid secretion after immunoprecipitation with mucin-specific antibodies. These studies have shown that membrane-associated mucins are produced in both parotid and submandibular glands and that they are expressed in different cell types than gel-forming mucins. Although the function of these mucins in the oral cavity remains to be elucidated, it is possible that they both contribute to the epithelial protective mucin layer and act as receptors initiating one or more intracellular signal transduction pathways.

Blotting, Northern↗

[Clinical analysis to 36 cases of acute laryngeal damage].

OBJECTIVE: To research the way to deal with the acute laryngeal damage. METHOD: Data of 36 cases with acute laryngeal damage were analyzed. RESULT: Among 36 cases, there were 10 cases who received non-surgical treatment and their phonation was normal except 2 cases who had mild hoarseness. Among 26 cases who had been performed operation. 21 had normal phonation after operation. 5 cases still had roughness of voice. There were 22 cases with normal respiration but 4 cases with larynostenosis and difficulty in decannulation. Among these 4 cases, 2 cases were decannulated respectively after 2 months and 10 years after operation. 29 cases made of 80% in all were cured and decannulated within 1 month. CONCLUSION: Acute laryngeal damage patients should be diagnosed and treated as early as possible according to the damage degree so as to recovery the laryngeal and tracheal's function and to prevent the sequelae and to increase the cure rate.

Acute Disease↗

A Cycloaddition Approach toward the Synthesis of Substituted Indolines and Tetrahydroquinolines.

The intramolecular Diels-Alder reaction of 2-substituted aminofurans (IMDAF) results in the formation of various indolines and tetrahydroquinolines. The isolation of these ring systems from the IMDAF reaction can be rationalized in terms of an initial [4 + 2]-cycloaddition that first produces an oxa-bridged cycloadduct, which was not detected since it readily underwent nitrogen-assisted ring opening. Proton exchange followed by an eventual dehydration provides the aromatic product. In certain cases, the intermediate cyclohexadienol can be isolated and independently converted to the final product in high yield. The starting 2-aminofurans were readily prepared from furanyl acyl azide by a Curtius rearrangement in the presence of an alcohol. Alkylation of the resulting N-alkyl carbamate with an alkenyl bromide allows for the synthesis of a wide variety of cycloaddition precursors. The scope of the IMDAF reaction was evaluated by using mono- and disubstituted alkenes, electron rich and electron deficient olefins, and acetylenic tethers. Cyclic 2-amidofurans were also synthesized using a related intramolecular Diels-Alder reaction of 2-amido-substituted oxazoles which contain a tethered alkyne. This transformation represents a new route to this rare heterocyclic ring system. The sequential cycloaddition method was used for a formal synthesis of the pyrrolophenanthridone alkaloid hippadine.

Journal Article↗

Anxiolytic agent, dihydrohonokiol-B, recovers amyloid beta protein-induced neurotoxicity in cultured rat hippocampal neurons.

The effects of anxiolytic honokiol derivative, dihydrohonokiol-B (DHH-B), on amyloid beta protein (Abeta(25-35), 10 nM)-induced changes in Cl(-)-ATPase activity, intracellular Cl- concentration ([Cl-]i) and glutamate neurotoxicity were examined in cultured rat hippocampal neurons. DHH-B (10 ng/ml) recovered Abeta-induced decrease in neuronal Cl(-)-ATPase activity without any changes in the activities of Na+/K+-ATPase and anion-insensitive Mg2+-ATPase. A GABA(C) receptor antagonist (1,2,5,6,-tetrahydropyridin-4-yl) methyl-phosphinic acid (TPMPA, 15 microM), inhibited the protective effects of DHH-B on Cl(-)-ATPase activity. DHH-B reduced Abeta-induced elevation of [Cl-]i as assayed using a Cl(-)-sensitive fluorescent dye, and prevented Abeta-induced aggravation of glutamate neurotoxicity. These data suggest that DHH-B exerts the neuroprotective action against Abeta through GABA(C) receptor stimulation.

Adenosine Triphosphatases↗