Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0001511 (Adhesion)
 5,955 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Kalinin was purified from squamous cell carcinoma (SCC25) spent culture media using an immunoaffinity column prepared from the mAb BM165. The affinity-purified material was separated by SDS-PAGE into three bands of 165-155, 140, and 105 kD identical to those obtained from normal human keratinocyte cultures and previously identified as kalinin. Kalinin promoted adhesion of a large number of normal cells and established cell lines with an activity similar to other adhesion molecules such as the laminin-nidogen complex, fibronectin, or collagen IV. However, kalinin was a much better substrate than laminin-nidogen complex for adhesion of cells of epithelial origin including primary human keratinocytes. Adhesion to kalinin was followed by cell shape changes ranging from rounded to fully spread cells depending on the cell types. The adhesion-promoting activity of kalinin was conformation dependent and was abolished by heat denaturation. mAb BM165 prevented cell adhesion to kalinin but not to other extracellular matrix substrates. However, either complete or partial inhibition was observed with different cells suggesting the existence of at least two cell-binding sites on the kalinin molecule. Experiments inhibiting cell adhesion with function-blocking anti-integrin subunit antibodies indicated that both alpha 3 beta 1 and alpha 6 beta 1 integrins are involved in the cellular interactions with kalinin, while for cell adhesion to classical mouse Engelbreth-Holm-Swarm laminin only alpha 6 beta 1 integrins, and not alpha 3 beta 1, appeared to be functional. Altogether, these results suggest that kalinin may fulfill additional functions than laminin, particularly for epithelial cells.
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PMID:Kalinin is more efficient than laminin in promoting adhesion of primary keratinocytes and some other epithelial cells and has a different requirement for integrin receptors. 813 72

Recent studies of spinal cord development and plasticity, in chick, have demonstrated a loss of regenerative ability correlating to embryonic day (E) 13 of the 21-day developmental period. Here we describe membrane fractions from embryonic chick spinal cords as permissive or restrictive substrates for the neuron-like differentiation of neuroblastoma x glioma hybrid NG108-15 cells, in vitro. Plasma membranes were purified from the thoracic spinal cord of embryos at a series of developmental stages (E10-E18). Micro-well plates were coated with the fractions and NG108-15 cells cultured thereon. Cells adhered to the E10-coated wells and began to differentiate after 2 h, becoming highly differentiated, with neurites 2-3 times longer than the diameter of the cell body after 24 h in in culture. In contrast, cells cultured in E18-coated wells remained as clusters of undifferentiated cells of rounded morphology, even after 48 h in culture. As well, the permissive and restrictive plasma membranes were assessed semiquantitatively as the number of adhering cells after 20 h of culture. Adhesion of cells to the substrate decreased as the embryonic age of the plasma membrane substrate increased. Examination of the plasma membrane fractions, using SDS-PAGE, revealed several proteins in the 40-60 kDa range that varied substantially between E12, E14 and E18. Results of this study provide in vitro confirmation of previous in vivo findings; namely, that early embryonic spinal cord is initially permissive for neuritic outgrowth becoming restrictive around E13.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Developmental transition by spinal cord plasma membranes of embryonic chick from permissive to restrictive substrates for the morphological differentiation of neuroblastoma x glioma hybrid NG108-15 cell. 845 60

Pseudomonas aeruginosa causes a variety of diseases in humans including lung and ocular infections. Infections of the cornea are usually associated with wearing contact lenses and can result in loss of vision. This study aimed to determine the effect of carbon or nitrogen limitation on the adhesion to contact lenses of a strain of Ps. aeruginosa isolated from contact lens-related corneal inflammation. Cells were grown in a continuous culture apparatus in varying levels of glucose or ammonia to effect nutrient limitation. Adhesion to contact lenses was measured as total counts and viable counts. The cell surface hydrophobicity and charge were measured using adhesion to surface-modified Sepharose. Changes in lipopolysaccharide were determined using 1D SDS-PAGE and changes in cell-surface proteins were measured using 2D gel electrophoresis. The more the cultures were nitrogen limited, the greater the increase in adhesion to unworn hydrogel contact lenses 0.3 x 10(3) - 2.2 x 10(3) cells/mm2 on Etafilon A lenses. Cells that were carbon limited showed a greater increase in adhesion to contact lenses when the lenses had been coated in artificial tears. It appeared that lipopolysaccharide may have been involved in the constitutive adhesion to unworn lenses that occurred during C-limitation, whereas changes in the outer membrane proteins contributed to the increased adhesion under nitrogen limitation, or the change in adhesion that occurred to carbon-limited cells using contact lenses coated in artificial tears. Nine cell-surface proteins appeared during nitrogen limitation with kDa/pI of 75/4.8, 4.9, 5.0; 62/5.6; 89/6.5; 38/6.4; 28/1.5; 18/6.4; 12/4.5. Any or all of these may have been involved in the increased adhesion and further experiments are underway to examine this possibility.
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PMID:Effect of nutrient limitation on adhesion characteristics of Pseudomonas aeruginosa. 1038 43

Streptococcus mutans is a major etiological agent in dental caries. Salivary agglutinin is one of the main salivary components binding to S. mutans. To learn more about the interaction of salivary agglutinin with S. mutans, parotid, submandibular, sublingual and palatal saliva samples were incubated with S. mutans suspension. Both depleted saliva samples and bacterial extracts were analyzed by SDS-PAGE and immunoblotting. Salivary agglutinin was present in all types of glandular saliva and in all cases bound to S. mutans, also to PC337C, a P1 mutant of S. mutans. Agglutinin was separated by SDS-PAGE under reducing and non-reducing conditions and then transferred to nitrocellulose. Non-reduced agglutinin bound S. mutans, but reduced agglutinin did not. Adhesion of S. mutans to agglutinin-coated microplates was inhibited by amine-containing components, 1 M NaCl or KCl and EDTA. Adhesion decreased with decreasing pH with no adhesion below pH 5.0. These data suggest that calcium-dependent electrostatic interactions play a role in binding. By immunoblotting was demonstrated that blood group antigens and Lewis antigens were present on agglutinin. Synthetic blood group antigens and Lewis antigens covalently coupled to polyacrylamide were tested for binding to S. mutans. Only Le(a)(Gal beta 1,3(Fuc alpha 1,4)GlcNAc) bound to S. mutans, whereas the blood group antigens Le(b), Le(x), Le(y), H1, H2, A, B and sialylated Le(a) did not. Lea without galactose (Fuc alpha 1,4GlcNAc) still bound to S. mutans, but Le(a) without fucose (Gal beta 1,3GlcNAc) did not. Binding of agglutinin to S. mutans was not inhibited by Le(a). In conclusion, S. mutans can bind to Le(a) carbohydrate epitopes in which the fucose is an essential residue. Le(a) carbohydrate epitopes are present on salivary agglutinin but play no major role in binding.
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PMID:A role for Lewis a antigens on salivary agglutinin in binding to Streptococcus mutans. 1069 74

Adhesion of mature asexual stage Plasmodium falciparum parasite-infected erythrocytes (iRBC) to the vascular endothelium is a critical event in the pathology of Plasmodium falciparum malaria. It has been suggested that the clag gene family is essential in cytoadherence to endothelial receptors. Primers used in PCR and RT-PCR assays allowed us to determine that the gene encoding CLAG 3 (GenBank accession no. NP_473155) is transcribed in the Plasmodium falciparum FCB2 strain. Western blot showed that antisera produced against polymerized synthetic peptides from this protein recognized a 142-kDa band in P. falciparum schizont lysate. Seventy-one 20-amino-acid-long nonoverlapping peptides, spanning the CLAG 3 (cytoadherence-linked asexual protein on chromosome 3) sequence were tested in C32 cell and erythrocyte binding assays. Twelve CLAG peptides specifically bound to C32 cells (which mainly express CD36) with high affinity, hereafter referred to as high-affinity binding peptides (HABPs). Five of them also bound to erythrocytes. HABP binding to C32 cells and erythrocytes was independent of peptide charge or peptide structure. Affinity constants were between 100 nM and 800 nM. Cross-linking and SDS-PAGE analysis allowed two erythrocyte binding proteins of around 26 kDa and 59 kDa to be identified, while proteins of around 53 kDa were identified as possible receptor sites for C-32 cells. The HABPs' role in Plasmodium falciparum invasion inhibition was determined. Such an approach analyzing various CLAG 3 regions may elucidate their functions and may help in the search for new antigens important for developing antimalarial vaccines.
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PMID:Identifying Plasmodium falciparum cytoadherence-linked asexual protein 3 (CLAG 3) sequences that specifically bind to C32 cells and erythrocytes. 1565 79

Atypical enteropathogenic Escherichia coli (aEPEC) strains are frequently implicated in infant diarrhoea in developing countries. Not much is known about the adherence properties of aEPEC; however, it has been shown that these strains can adhere to tissue-cultured cells. A chromosomal region designated the locus for diffuse adherence (LDA) confers aEPEC strain 22 the ability to adhere to culture cells. LDA is an afimbrial adhesin that contains a major subunit, LdaG, whose expression is induced on MacConkey agar at 37 degrees C. We hypothesized that the bile salts found in this culture media induce the expression of LdaG. Strain 22 and the LdaG mutant were grown in Luria-Bertani (LB) media in the presence or absence of bile salts and heat-extracted surface-expressed proteins were separated by SDS-PAGE to determine whether expression of the 25 kDa LdaG protein was induced. Western blot analysis with anti-LdaG confirmed that bile salts enhance LdaG expression at 37 degrees C. Adhesion assays on HeLa cells revealed that adhesion in a diffuse pattern of strain 22 increased in the presence of bile salts. We also confirmed that expression of the localized adherence pattern observed in the ldaG mutant required the presence of a large cryptic plasmid found in strain 22 and that this phenotype was not induced by bile salts. At the transcriptional level, the ldaG-lacZ promoter fusion displayed maximum beta-galactosidase activity when the parent strain was grown in LB supplemented with bile salts. Fluorescence Activated Cell Sorting analysis, immunogold labelling electron microscopy and immunofluorescence using anti-LdaG sera confirmed that LDA is a bile salts-inducible surface-expressed afimbrial adhesin. Finally, LdaG expression was induced in presence of individual bile salts but not by other detergents. We concluded that bile salts increase expression of LDA, conferring a diffuse adherence pattern and having an impact on the adhesion properties of this aEPEC strain.
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PMID:Bile salts induce expression of the afimbrial LDA adhesin of atypical enteropathogenic Escherichia coli. 1738 33

Balamuthia amoebic encephalitis (BAE) is a serious human disease almost always leading to death. An important step in BAE is amoebae invasion of the bloodstream, followed by their haematogenous spread. Balamuthia mandrillaris entry into the central nervous system most likely occurs at the blood-brain barrier sites. Using human brain microvascular endothelial cells (HBMECs), which constitute the blood-brain barrier, this study determined (i) the ability of B. mandrillaris to bind to HBMECs and (ii) the associated molecular mechanisms. Adhesion assays revealed that B. mandrillaris exhibited greater than 90 % binding to HBMECs in vitro. To determine whether recognition of carbohydrate moieties on the surface of the HBMECs plays a role in B. mandrillaris adherence to the target cells, adhesion assays were performed in the presence of the saccharides mannose, galactose, xylose, glucose and fucose. It was observed that adherence of B. mandrillaris was significantly reduced by galactose, whilst the other saccharides had no effect. Acetone fixation of amoebae, but not of HBMECs, abolished adhesion, suggesting that B. mandrillaris adhesin(s) bind to galactose-containing glycoproteins of HBMECs. B. mandrillaris also bound to microtitre wells coated with galactose-BSA. By affinity chromatography using a galactose-Sepharose column, a galactose-binding protein (GBP) was isolated from detergent extracts of unlabelled amoebae. The isolation of a GBP from cell-surface-biotin-labelled amoebae suggested its membrane association. One-dimensional SDS-PAGE confirmed the proteinaceous nature of the GBP and determined its molecular mass as approximately 100 kDa. This is the first report suggesting the role of a GBP in B. mandrillaris interactions with HBMECs.
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PMID:Balamuthia mandrillaris interactions with human brain microvascular endothelial cells in vitro. 1764 21

Adhesion of microorganisms to dental surfaces is the initial step in the formation of dental bacterial plaque. Streptococcus mutans (S. mutans) is considered the main causal agent of one of the most common diseases in humans: dental caries. Adherence of these bacteria results from the interaction of adhesins that form part of their structure with salivary components, specifically those that compose the acquired pellicle. The complexity of this interaction has been the subject of studies in past years, to the extent of identifying certain salivary components related to adhesion to enamel surfaces, such as proline-rich proteins (PRSs), Staherins, Histatins, Cystatins, etc. One of the objectives of this study was to determine the adhesion capacity of S. mutans to synthetic hydroxyapatite incubated with saliva samples of caries-active and caries-inactive individuals. For the purpose of these assays, both the whole saliva samples and the salivary protein extracts were used. They were obtained by separating the proteins contained in the simple SDS-PAGE, in three ranges of molecular weight, selected in accordance with the electrophoresis profile that was usually found. The results indicated that the adhesion of this microorganism was greater in caries-inactive patients, when tested with whole saliva and proteins in the 120-159 kDa molecular weight range. This suggests that adhesion, per se, does not have a definite effect on the mechanisms that cause the disease in some individuals. However, these are interesting findings that may contribute to the design of strategies to control the adhesion of S. mutans to the tooth's surface.
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PMID:Adhesion of Streptococcus mutans to salivary proteins in caries-free and caries-susceptible individuals. 1764 12

The fed operon gene clusters with each size of 5.6kb, encoding the F18ab or F18ac fimbriae, was amplified respectively by high fidelity PCR using the genomic DNA templates from F18 fimbriae E. coli strains 107/86 or 2134P. The PCR products with the restriction enzyme sites at each end were digested and then cloned into the vector pET-22b (+), the recombinant plamids with the inserts of both type of fed gene clusters were constructed and screened, further confirmed by the means of combination with restriction endonuclease analysis and sequencing. The both types of fimbriae F18ab and F18ac were expressed efficiently in the E. coli BL21 (DE3) after proper concentration of IPTG induction. Expressed fimbriae were revealed and confirmed by transmissible electromicroscope observation. The both fimbriae F18ab and F18ac were isolated and purified from the recombinant E. coli, and only a single major band of protein with size of approximately 15kDa was visualized in Coomassie blue-stained gels after SDS-PAGE. The rabbits sera with high titer of anti-F18 fimbriae were detected after being immunized with the purified F18ab or F18ac fimbriae. The results of combination of agglutination assay with Western blotting showed that the sera directed against both fimbriae F18ab and F18ac reacted positively with the F18 fimbriae from both wild E. coli 107/86 and 2134P. Small intestine epithelial cells with F18 fimbriae receptors, which were from post-weaning piglets with the genotypes of FUT1 gene both M307(GG) and M307(AG), were prepared and tested for the adherence of E. coli expressing F18 fimbriae under the microscopic examination. Adhesion and adhesion inhibition test showed both of the recombinant E. coli expressing F18ab or F18ac fimbriae respectively could adhere to the jejunal epithelial cells in vitro as E. coli 107/86 and 2134p did. The both of anti-sera directed against fimbriae F18ab or F18ac respectively can efficiently inhibit the fimbriae-mediated post-weaning piglet jejunal epithelial cells adherence to both the recombinant E. coli (expressing F18ab or F18ac fimbriae) and wild type E. coli (107/86 and 2134P).
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PMID:[Cloning and expression of F18 fimbrial operon gene clusters from enterotoxigenic Escherichia coli and their bioactivity]. 1806 50

Four different colony morphologies were produced by Flavobacterium columnare strains on Shieh agar plate cultures: rhizoid and flat (type 1), non-rhizoid and hard (type 2), round and soft (type 3), and irregularly shaped and soft (type 4). Colonies produced on AO agar differed from these to some extent. The colony types formed on Shieh agar were studied according to molecular characteristics [Amplified Fragment Length Polymorphism (AFLP), Automated Ribosomal Intergenic Spacer Analysis (ARISA), and whole cell protein SDS-PAGE profiles], virulence on rainbow trout fingerlings, and adhesion on polystyrene and fish gills. There were no molecular differences between colony types within one strain. Type 2 was the most adherent on polystyrene, but type 1 was the most virulent. Adhesion of F. columnare strains used in this study was not connected to virulence. From fish infected with colony type 1, three colony types (types 1, 2 and 4) were isolated. Contrary to previous studies, our results suggest that strong adhesion capacity may not be the main virulence factor of F. columnare. Colony morphology change might be caused by phase variation, and different colony types isolated from infected fish may indicate different roles of the colony morphologies in the infection process of columnaris disease.
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PMID:Flavobacterium columnare colony types: connection to adhesion and virulence? 1898 35


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