Biomedical subjects

T Wadström

Publications and source records attributed to T Wadström.

At least 145 records · Page 8Linked to original sources

PubMed1993

Binding of Escherichia coli to Penrose rubber drains--an in vitro study.

Ten different isolates of Escherichia coli were used to compare bacterial attachment to Penrose rubber drains at different temperatures and to investigate a possible relation with cell surface hydrophobicity and charges, as well as the capacity of autoaggregation. Penrose rubber drain pieces of 1 cm2 were incubated with 4.8 x 10(6) E. coli cells for 1 h at 22, 37 and 42 degrees C, respectively. After rinsing with phosphate buffered saline (PBS), the number of adhering bacteria on the drain pieces was calculated by measuring cellular ATP (adenosine triphosphate) bioluminescence. Bacterial cell surface properties at different temperatures were determined by two-phase partitioning and autoaggregation was determined by arbitrary scoring under the dissecting microscope. For most strains of E. coli, the number of adherence to Penrose rubber drain was higher at 22 degrees C than at 37 degrees C and 42 degrees C. Bacterial cell-surface properties and autoaggregation capacity were influenced by the growth temperature, but without correlation to bacterial ability to attach to rubber drains. Thus, the present study demonstrates that attachment of E. coli to Penrose rubber drains was significantly influenced by temperature, but bacterial cell-surface hydrophobicity and charge as well as autoaggregation capacity had no influence on bacterial attachment ability.

Bacterial Adhesion↗

PubMed1993

Binding of collagen, fibronectin, lactoferrin, laminin, vitronectin and heparan sulphate to Staphylococcus aureus strain V8 at various growth phases and under nutrient stress conditions.

We have examined how Staphylococcus aureus strain V8 cells interact with 125I-labelled extracellular matrix (ECM) and serum proteins (collagen type I and IV), fibronectin, lactoferrin, laminin, vitronectin, and heparan sulphate at various phases of the growth cycle. Maximal binding of these glycoproteins and heparan sulphate to the bacteria occurred after 17 to 20 h in the late stationary phase except for fibronectin-binding, which was maximal after 12 to 14 h. Binding of the glycoproteins and heparan sulphate to S. aureus V8 under nutrient stress conditions exhibited complex patterns based on different starving conditions and various binding ligands. In general, bacteria starved in distilled water and 0.02 M potassium phosphate buffer (pH 7.2) at room temperature showed high susceptibility to all binding ligands within the first 18 h, followed by entering a lower binding period (except for collagen-binding which still remained high). The binding was not correlated to cell surface charge or hydrophobicity of the bacteria. Furthermore, extracellular and cell-associated proteolytic activity of starved cells against ECM and serum proteins was found to be greater than for non-starved cells. Thus, S. aureus could sustain its ability to bind various connective tissue and cell surface components during a long period of time even in the absence of energy-yielding substrates.

Blood Proteins↗

PubMed1993

Studies on binding of glycosaminoglycans to Streptococcus pyogenes by using 125I-heparan sulphate as a probe.

Binding of 125I-heparan sulphate to the cell surface of Streptococcus pyogenes is mediated by proteins, that could be released from the streptococcal cell wall by using alkaline buffer. SDS-electrophoresis revealed two bands with molecular weights of 63 and 58 kDa. Binding of the 125I-labelled heparan sulphate probe to streptococci seems to be due to charge interactions, as the same probe was displaced by unlabelled heparan sulphate, other negatively charged molecules such as heparin, dextran sulphate, dermatan sulphate or by high ionic strength. The interaction was also strongly influenced by pH. The binding constant at pH 7.2 was estimated to be 9.8 x 10(6) mol/l, suggesting a moderate affinity. The presence of collagen of different types enhanced binding of 125I-labelled heparan sulphate to streptococci, whereas fibronectin and vitronectin had an inhibitory effect. The cooperation between heparan sulphate and collagen could be important for the adhesion of streptococci to connective tissue.

Bacterial Outer Membrane Proteins↗

PubMed1993

Isolation of a sialic acid-specific surface haemagglutinin of Helicobacter pylori strain NCTC 11637.

A deionized water extract of Helicobacter pylori NCTC 11637 contained haemagglutinin activity that was (i) soluble (i.e., not associated with particulate material sedimented by centrifugation at 100,000 x g for 1 h), (ii) stable to lyophilization, (iii) heat-labile, (iv) chymotrypsin-sensitive, (v) inhibited by fetuin, orosomucoid, and NANLac, but not by asialofetuin and (vi) inactive against guinea pig erythrocytes incubated with Clostridium perfringens neuraminidase, but active against untreated guinea pig erythrocytes. The data support the idea that the haemagglutinin is a protein which recognizes the alpha-(2-3) structure of sialylated glycoconjugates. Fractionation of the extract by isoelectric focusing and by gel filtration with Sephacryl S-400 indicated that the haemagglutinin has a pI of 3.7 and consist of high molecular-weight-protein aggregates. SDS-PAGE analysis of the preparation purified by gel filtration showed 3 protein bands at ca. 64 kD, 56 kD and 20 kD. Electron microscopy of H. pylori incubated with gold-labelled fetuin indicated that the haemagglutinin was associated with loosely adherent material on the bacterial surface, and that the purified haemagglutinin did not reveal a fimbrial structure. The ability to bind to sialoglycoconjugates on the erythrocyte membrane suggests that the haemagglutinin may be an important colonization factor enabling H. pylori to bind to similar saccharide structures on epithelial cells.

Animals↗

PubMed1993

Adherence of haemagglutinating Helicobacter pylori to five cell lines.

Helicobacter pylori causes gastritis and is an important factor for the development of peptic ulcer disease in man. We used two different methods to examine the adhesion of nine H. pylori strains, with different haemagglutinating properties, to five cell lines, HeLa S3, HFI, Vero, SW1222 and WEHI cells. The adhesion studies were performed a) as bacterial adhesion to a monolayer of tissue culture cells, visualizing bacteria with fluorescein-isothiocyanate-labelled antibodies, b) as cell agglutination with bacteria and eucaryotic cells mixed in a suspension. The H. pylori strains were divided into three groups according to their cell adhesion properties. In general, H. pylori strains which showed the best adhesion to the five cell lines were strains which showed the best capability of agglutinating erythrocytes of several animal species. It is likely that the same adhesins are involved in cell adhesion and in haemagglutination. The two methods gave similar results.

Animals↗

PubMed1993

Affinity of the gastric pathogen Helicobacter pylori for the N-sulphated glycosaminoglycan heparan sulphate.

Binding of 125I-heparan sulphate was a common property of Helicobacter pylori strains isolated from patients with gastroduodenal ulcer diseases. Binding was (i) saturable; (ii) reversible by the addition of unlabelled heparan sulphate and heparin; (iii) inhibited by unlabelled heparan sulphate, heparin, and heparin oligosaccharides but not by other glycosaminoglycans of comparable size (chondroitin sulphate and dermatan sulphate) or by highly glycosylated glycoproteins (hog gastric mucin and fetuin); (iv) reduced by heat treatment (80 degrees C, 10 min) and exposure of the bacteria to pronase E, proteinase K, trypsin and chymotrypsin, but unaffected by treatment with pepsin and neuraminidase; and (v) time-, pH-, and ionic strength-dependent. Scatchard plot analysis of the binding data indicated the presence of one class of high-affinity receptor (Kd = 9 x 10(-9) M) for heparan sulphate.

Binding, Competitive↗

PubMed1993

High-affinity binding of the basement membrane protein collagen type IV to the crystalline virulence surface protein array of Aeromonas salmonicida.

The surface of the fish pathogen Aeromonas salmonicida is covered by a paracrystalline array (the A-layer) which is a virulence factor for the organism. Quantification of the ability of A. salmonicida cells to bind collagen types I and IV in a 125I-radiolabelled liquid-phase assay showed that A-layer-positive cells bound high levels of collagen type IV, but significantly lower levels of collagen type I. Collagen type IV binding was confirmed using non-radiolabelled enzyme-linked immunosorbent assays. 125I-Collagen type IV binding was rapid, specific, saturable, high affinity, and essentially irreversible by unlabelled collagen type IV. The A-layer was responsible for collagen type IV binding because binding was inactivated by selective removal of the A-layer at pH 2.2, and neither isogenic A-layer-deficient A. salmonicida mutants nor strains of Aeromonas hydrophila possessing a morphologically similar paracrystalline array bound this basement membrane protein.

Aeromonas↗

PubMed1993

Protective opsonic activity of antibodies against fibronectin-binding proteins (FnBPs) of Staphylococcus aureus.

In this report, opsonic activity of hyperimmune rabbit IgG against fibronectin-binding proteins (gal-FnBP A and ZZ-FnBP B) of Staphylococcus aureus is described. Moreover, the action of IgG purified from serum of rabbits immunized with 'combined vaccine' (fibronectin-binding protein A+collagen-binding protein+alpha-toxoid) is shown. The opsonic activity has been studied in an in vitro phagocytosis assay as well as in vivo. Mice which had been infected intraperitoneally with S. aureus strain Cowan 1 pretreated (opsonized in vitro) with specific anti-FnBPs IgG were able to eliminate the staphylococci from the peritoneal cavity and liver more rapidly than controls. Also, clearance from the bloodstream of intravenously injected S. aureus Cowan 1 as well as S. aureus U320, opsonized with IgG anti-FnBPs or anti-FnBP+CnBP+alpha-toxoid, was more effective than observed in control groups. In other in vivo experiments it was shown that mice passively immunized with hyperimmune IgG anti-FnBP (one or two doses, intravenously) before challenge with S. aureus Cowan I eliminated the bacteria better than controls injected only with preimmune IgG.

Adhesins, Bacterial↗

PubMed1993

High-affinity binding of laminin by Helicobacter pylori: evidence for a lectin-like interaction.

Laminin, the major glycoprotein of basement membranes, was shown to be bound by the human gastric pathogen Helicobacter pylori. Binding of 125I-laminin by strain 17874 was time-dependent, specific and saturable. Scatchard analysis of specific binding indicated about 2000 binding sites per cell with a dissociation constant of 8.5 pM. Treatment of the cells by heat (80 degrees) and with proteolytic enzymes drastically reduced laminin binding, suggesting that the laminin receptors are surface proteins. Some highly glycosylated glycoproteins inhibited laminin binding by 50%. Furthermore, N-acetylneuraminyllactose decreased laminin binding by 70% and neuraminidase treatment of laminin by 50%, while a recombinant B1 chain of laminin, containing high-mannose type oligosaccharides, inhibited binding by only 25%. This suggests that terminal sialic acids on laminin compete for a specific sugar binding protein(s) on H. pylori cells.

Bacterial Adhesion↗

PubMed1993

Interferon-gamma, interleukin-1 and tumour necrosis factor-alpha synthesis during experimental murine staphylococcal infection.

Several exotoxins of Staphylococcus aureus were shown to modulate the host immune system by stimulation of monokine release. BALB/c mice infected intravenously (i.v.) with live cells if S. aureus, strain Cowan 1, had a detectable serum level of TNF-alpha at 3, 4 and 5 h after injection. When S. epidermidis (strain E3380, clinical isolate) was used to infect mice, the level of TNF-alpha was lower (the detection limit of the cytotoxicity assay with WEHI cells was 40 pg ml-1). Kinetics of TNF synthesis was different from that observed in experimental infections caused by Gram-negative bacteria. Similarly to TNF-alpha, IL-1 alpha appears in a measurable level at 3 h after i.v. injection of bacteria. The highest serum level of IFN-gamma was observed 12 h after infection with both S. aureus and S. epidermidis. A quantity ten times more of S. epidermidis than of S. aureus cells was required to induce similar levels of TNF-alpha and IFN-gamma. Recombinant IFN-gamma administered in vivo in four daily doses followed by infection of S. aureus resulted in increased elimination of bacteria from the spleen, liver and peritoneal cavity of mice.

Animals↗

PubMed1993

Reduction of E. coli adherence to rubber slices treated with phospholipids.

The present study aimed at modifying the surface of biliary drain material to reduce bacterial adherence. The adherence of cells of seven E. coli strains to rubber slices treated with phosphatidylcholin (PC) or phosphatidylinositol (PI) and the adherence of cells of E. coli strain NG7C to PC- or PI-treated rubber slices implanted in the common bile duct in rats were studied in vitro. The rubber slices were incubated with 1 x 10(7) cfu radiolabeled E. coli cells/ml at 37 degrees C for 60 min and then drained and washed thrice in 2 ml PBS, and adherent E. coli cells were quantified by radioactivity counting. The results show that both PC and PI absorbed on the surface of slices reduced the adherence of E. coli cells in at least two ways, i.e. by changing surface properties in vitro and by reducing deposition of host-derived molecules on phospholipid-treated surfaces in vivo. The results may be of use for modification of the biomaterial surface in the clinical situation.

Animals↗

PubMed1993

Rapid detection and characterization of sialic acid-specific lectins of Helicobacter pylori.

A particle agglutination assay (PAA) using fetuin (Ft) covalently coupled to carboxylate-modified latex (CML) particles was evaluated for rapid detection of sialic acid-specific haemagglutinins/lectins (SALs) of Helicobacter pylori isolates which bind sialoglycoconjugates. Sixty-three percent (20/32) of the isolates examined gave a positive PAA test. Cell-bound SALs were extracted by washing the bacteria with deionized water or isotonic saline, and their expression was influenced by pH and culture conditions. The Ft-CML reactivity of the PAA-positive isolates was inhibited by bovine submaxillary mucin, transferrin, fetuin, orosomucoid, vitronectin and lactoferrin in a manner which suggested that the isolates contain a lectin recognizing the alpha(2-6) linkage of terminal sialic acid. Western blots of strain NCTC 11637 SALs probed with horseradish peroxidase (HRP)-labelled Ft identified three bands (MW 64 kD, 62 kD, 56 kD) which also reacted with HRP-labelled mucin, transferrin, lactoferrin, orosomucoid, vitronectin and laminin. Sera from patients with a H. pylori infection and one polyclonal rabbit antiserum (strain NCTC 11637) also reacted with the SALs. Immunogold labelling of a polyclonal rabbit antiserum raised against the 64 kD protein of strain NCTC 11637 that reacted strongly with Ft-CML showed that abundant SALs were loosely cell-associated with the cell surface of both spiral and coccoidal forms of H. pylori. SALs were also present in low amounts on the surface of strain NCTC 11638 and 66, a clinical isolate that did not react with Ft-CML.

Animals↗

PubMed1993

The expression of potential colonization factors of yeasts isolated from fish during different growth conditions.

Three strains, Rhodotorula rubra, Rhodotorula glutinis, and Candida zeylanoides, isolated from fish, were tested for the expression of putative tissue colonization factors. All strains were able to bind collagen type I, fibronectin, and laminin to various degrees after growing on various solid and broth media, while the binding to collagen type IV was sparse under all conditions tested. For the three strains tested, a very low cell surface hydrophobicity was shown for growth on various solid and broth media. Mostly, the strains also expressed a negatively charged surface. Extracellular protease activity using different substrates was shown for all three strains. Furthermore, two properties related to iron scavenging, i.e., binding of lactoferrin and production of siderophores, were also tested. For the three strains a capacity to bind lactoferrin as well as a capacity to excrete siderophores were demonstrated. Since these different properties have been correlated to virulence and to the capacity of colonization in other organisms, we address the question of whether the expression of these properties in yeasts could contribute to colonization in fish.

Animals↗

PubMed1992

Surface properties, connective tissue protein binding and Shiga-like toxin production of Escherichia coli isolated from patients with ulcerative colitis.

Escherichia coli strains isolated from intestinal biopsies of patients with ulcerative colitis (n 146), Crohn's disease and colonic polyposis (n 41) were analysed for binding of collagen I, collagen IV, fibronectin and laminin. Strains expressed varying degrees of binding of one or more of the four connective tissue proteins. Only 32 strains did not express binding of any of the proteins. The strains expressed low or moderate cell surface hydrophobicity. There was no correlation between protein binding and expression of cell surface hydrophobicity. E. coli isolated from inflamed rectal mucosa were slightly less negatively charged than strains isolated from healthy intestinal mucosa. Shiga-like toxins I and II were detected in 32 strains from 28 patients. Of these, 5 strains had been isolated from normal or healed tissue. In patients with inflammatory bowel disease, connective tissue proteins are exposed in intestinal ulcerations. Strains expressing binding of one or several of these proteins may have a selective advantage to colonize these lesions.

Adolescent↗

PubMed1992

Vitronectin-binding surface proteins of Staphylococcus aureus.

S. aureus strain ISP 546 was selected (of 55 strains tested) to define optimal conditions for expression of vitronectin binding. High binding was expressed when the strain was grown on blood agar and in Todd-Hewitt broth. Binding was optimal in the 6.0 to 7.2 pH range and was unaffected by divalent cations and ionic strength. Binding was partially inhibited by D-mannose, heparin, types I and IV collagen, fibronectin, fibrinogen and vitronectin, but was not affected by other carbohydrates or glycoproteins tested. Cell surface binding components were extracted with the aid of 1 M LiCl (pH 5.0) from strain ISP 546 grown in Todd Hewitt broth. Vitronectin binding proteins were purified by affinity chromatography on heparin-Sepharose. Fractions inhibiting binding of 125I-labelled vitronectin to strain ISP 546 were eluted by 0.01 M NaOH, dialysed, concentrated and subjected to SDS-PAGE. Silver staining revealed one major band (70 kDa) and two minor bands (34 and 36 kDa).

Binding, Competitive↗

PubMed1992

Binding of laminin, type IV collagen, and vitronectin by Streptococcus pneumoniae.

Forty-three strains of Streptococcus pneumoniae were tested for their ability to bind radiolabelled laminin, collagen types I, II and IV, fibronectin, and vitronectin. Two basement membrane components, laminin and type IV collagen, interacted with many S. pneumoniae strains. All strains bound laminin and 28 (65%) bound collagen type IV. Approximately 60% of the strains bound vitronectin but only a few strains showed low binding of fibronectin and collagen type I and II.

Bacterial Adhesion↗

PubMed1992

Binding of Chlamydia trachomatis serovar L2 to collagen types I and IV, fibronectin, heparan sulphate, laminin and vitronectin.

Chlamydia trachomatis serovar L2 elementary bodies were tested for binding to 125I-labelled components of the extracellular matrix. C. trachomatis bound high levels of collagen type I and heparan sulphate, whereas the binding to collagen type IV, fibronectin, laminin and vitronectin was substantially less. Preliminary characterization showed that chlamydial surface proteins are essential for binding to occur. Our results suggest that a selective affinity for C. trachomatis to a limited number of components of the extracellular matrix may have important implications mediating attachment to host cell surfaces and localization of chlamydiae in other tissues, such as joints.

Bacterial Adhesion↗

PubMed1992

Immunological response to a Staphylococcus aureus fibronectin-binding protein.

A protein (gal-FnBP), constructed by fusion of the genes encoding beta-galactosidase of Escherichia coli and the binding domains of fibronectin-binding protein (FnBP) of Staphylococcus aureus was used. FnBP is a surface protein responsible for attachment of bacteria to extracellular matrix of various host tissues. Gal-FnBP is more stable and can be produced in larger quantities than native FnBP. The binding specificity of this fusion protein was established in a Western blot analysis. Treatment of gal-FnBP with formalin inactivated the binding capacity of the protein but immunogenicity was retained. Immunisation of mice with formalin-treated gal-FnBP resulted in high antibody titres against the fibronectin-binding part of this fusion protein. These antibodies were measured by their ability to block the specific binding of fibronectin to gal-FnBP in a blocking assay. Sera raised against formalin-treated gal-FnBP and non-treated gal-FnBP blocked this binding to 40 and 25% respectively, thereby indicating the usefulness of gal-FnBP as a vaccine component.

Adhesins, Bacterial↗