Binding of purified Escherichia coli O75X adhesin to frozen sections of human kidney

Abstract Binding characteristics of the purified Escherichia coli O75X adhesin in frozen sections of human kidney were determined, using antibodies raised against the purified antigen and rhodamine-conjugated second antibodies. To identify the adhesin-binding nephron domains, the same tissue sections were double stained with fluorescein isothiocynate-conjugated nephron site-specific lectins. The results revealed that, at the tubular pole, the O75X adhesin bound selectively to the basement membrane of proximal and distal tubules and, with a slightly lower efficiency, of collecting ducts. In the glomerulus, the O75X adhesin bound only to the parietal epithelial cells (Bowman's capsule). The purified O75X adhesin bound also to exfoliated epithelial cells in human urine. Our results suggest that the O75X adhesin may contribute to the uropathogenicity of E. coli by binding the bacteria to tissue structures in the human urinary tract.     

Characterization of type 1 pili of Salmonella typhimurium LT2.

Type 1 pili from Salmonella typhimurium LT2 were purified and characterized. The pilus filaments were 6 nm in diameter and over 1 microns long. Estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight of the pilin was 21,000. The isoelectric point of the filament was 4.1. Hydrophobic amino acids comprised 40.3% of the total amino acids of the pilin, which contained more proline, serine, and lysine than reported for the type 1 pilin of Escherichia coli. Purified pili agglutinated both horse and chicken erythrocytes and yeast cells but not bovine, sheep, or human erythrocytes. Horse erythrocyte agglutination was inhibited at lower concentrations by alpha-methyl-D-mannoside than by yeast mannane and D-fructose. Agglutination was not affected by D-galactose or sucrose. Results of the present study confirm the role of type 1 pili as Salmonella hemagglutinins and show chemical differences between the type 1 pili of S. typhimurium and E. coli.     

Carbohydrate-rich tissue components in lung cancer and in normal bronchial tissues: a histochemical study

Synopsis The carbohydrate-rich compounds in bronchopulmonary neoplasms and in non-neoplastic tissue have been characterized histochemically. Glycogen was present in few epidermoid and large-cell carcinomas. Epithelial mucosubstances were produced by adeno-, mucoepidermoid, and large-cell carcinomas. The mucosubstances produced by carcinoma cells had characteristics different from those occurring in normal tissue. The most striking characteristic was the presence of a sialidase-labile compound in certain neoplasms.Hyaluronic acid was present in the stroma of the carcinomas. In a third of the cases studied, chondroitin sulphates were also present. Higher sulphated compounds were observed as well. This stromal reaction was correlated with the occurrence of a round cell reaction, but not with mast cells. This was considered to indicate the production of stromal material and fibres, but it can also explain the high levels of carbohydrate-rich substances in serum and urine in cases where neoplastic tissue itself does not produce such substances. It also agrees with the theory of carbohydrate-rich compounds acting as a barrier preventing immunological reactions against neoplastic cells.     

Histochemical demonstration of carbonic anhydrase activity in the alimentary canal

Summary Carbonic anhydrase (CAH) activity was histochemically demonstrated in various parts of the alimentary canal of rat and in the stomach of man using the method of Waldeyer and Häusler (1959). The most intense histochemical reaction was observed in the parietal cells of the rat stomach, and reactions of decreasing intensity in the epithelial cells of the colon, appendix, jejunoileum, duodenum and oesophagus in the order mentioned. An intense reaction was also observed in the parietal cells of the stomach of man and a weak activity in the pyloric glands. After electrophoresis on cellulose acetate film the CAH activity in human and rat stomach mucosa showed one band with the same migration rate as the fastest moving band of the erythrocyte CAH.     

Vegetation of the Special Zoological Reserve of Kopački Rit

This paper presents the results of several years' surveys on the vegetation of the Special Zoological Reserve of Kopaki Rit. Over this large floodplain of the Danube and Drava rivers prevail different types of hygro- and hydrophytic communities. Some of them are relatively constant, having persisted in this Reserve for many years with a relatively constant area. Some other communities, particularly aquatic ones, appear periodically, and their area is directly influenced by surface water area and duration of the flood.The vegetation can be divided into 36 plant communities within the classes Lemnetea, Potamogetonetea, Littorelletea, Phragmitetea, Isoëto-Nanojuncetea, Bidentetea tripartitae, Chenopodietea, Plantaginetea majoris, Molinio-Arrhenatheretea and Querco-Fagetea.     

Quantitative contributions of bacteria and of Deinococcus geothermalis to deposits and slimes in paper industry

Deinococcus geothermalis has frequently been isolated from pink colored deposits of paper industry processes. Laboratory studies have shown that D. geothermalis is capable of forming on nonliving surfaces patchy biofilms that are resistant to adverse agents such as extreme pH, desiccation, solubilising detergents and biocides. This study was done to quantitatively assess the role of D. geothermalis as a biofouler in paper industry. Colored deposits were collected from 24 European and North American paper and board machines and the densities of the bacterial 16S rRNA genes and those of the red slime producers D. geothermalis and Meiothermus spp. were measured by QPCR (quantitative real time PCR). D. geothermalis was found at nine machines, usually from splash area deposits, but its contribution was minor, 0.001–1%, to the total bacterial burden of 8.3 to log 10.5 log units per gram wet-weight of the deposits. When D. geothermalis was found in a measurable quantity, Meiothermus spp. also was found, often in bulk quantity (7–100% of the total bacteria). The data are in line with the properties of D. geothermalis known from laboratory biofilm studies, indicating this species is a pioneer coloniser of machine surfaces and may help other bacteria to adhere and grown into biofilms, rather than competing with them.     

Functional analysis of the minor subunits of S fimbrial adhesin (SfaI) in pathogenic Escherichia coli

S fimbrial adhesins I and II (SfaI and II), produced by extraintestinal Escherichia coli pathogens that cause urinary tract infections (UTI) and newborn meningitis (NBM), respectively, mediate bacterial adherence to sialic acid-containing glycoprotein receptors present on host epithelial cells and extracellular matrix. The S fimbrial adhesin complexes consist of four proteins: SfaI-A, the major subunit protein and the minor subunit proteins SfaI-G, SfaI-S and SfaI-H. Sialic acid-specific binding is mediated by the minor subunit protein SfaI-S. In order to determine whether the minor subunit proteins SfaI-G, -S and -H play a role in the modulation of adherence and the degree of fimbriation, a trans-complementation system was developed. A non-adhesive E. coli K-12 derivative, harbouring the sfaI-A gene but lacking sfaI-G, -S and -H, was transformed with sfaI-G, -S or -H. Only SfaI-S was able to increase the degree of fimbriation and to confer adhesion properties on the recombinant E. coli K-12 strains. Amino acid residues in SfaI-S that are involved in modulation of fimbriation as well as in receptor recognition were localized by random and site-directed mutagenesis.     

Immunohistochemical demonstration of carbonic anhydrase.

Rabbits were immunized using human erythroxyte carbonic anhydrase B (HCA B) purified by the modified methods of Armstrong et al. (1966) and Bernstein and Schraer (1972). The globulin fraction was isolated by ammonium sulphate precipitation. The anti-HCA B globulin was specific, when judged using the double diffusion technique of Ouchterlony and immunoelectrophoresis. No cross reaction with human erythrocyte carbonic anhydrase C was found, but cross reactions with erythrocyte carbonic anhydrase from rat, mouse and guinea pig were observed. Flurorescein isothiocyanate conjugated goat anti-rabbit globulin was used for the localization of HCA B in tissue sections and erythrocytes on slides.     

Adhesion of Escherichia coli to human kidney cryostat sections

Abstract A method for studying adhesion of fluorochrome-stained Escherichia coli to cryostat sections of human kidney is described. Of the 3 model strains used, the P-fimbriated strain KS71 adhered more effectively than the type-1-fimbriated or nonfimbriated strains. Adhesion of KS71 was inhibited specifically by anti-P-fimbriae Fab fragments, and it was concluded that P-fimbriae mediate E. coli adhesion in the kidneys.     

Characterization of dominant cultivable lactobacilli and their antibiotic resistance profiles from faecal samples of weaning piglets

AIMS: To examine the lactic acid bacteria flora of weaning piglets, to define the distribution of different lactobacilli species in piglet faecal samples, and to determine the susceptibility phenotype to 11 antibiotic of different families. METHODS AND RESULTS: The faecal samples were taken from piglets with good herd status at 11 and 28 days after weaning. The Lactobacillus isolates (n = 129) from 78 animals housed in pairs in 39 pens were preliminarily identified by their morphology and biochemical characteristics. Partial 16S ribosomal DNA (16S rDNA) was used to identify the isolates to the species level, and RAPD (randomly amplified polymorphism DNA) profiles to differentiate Lactobacillus isolates to the strain level. Based on these studies, 67 strains were selected for antibiotic resistant tests. The most numerous Lactobacillus species found in the piglets was Lactobacillus reuteri (n = 43). Other lactobacilli were L. salivarius (n = 15), L. agilis (n = 4), L. johnsonii (n = 2), L. vaginalis (n = 1), L. mucosae (n = 1) and L. gallinarum (n = 1). All the strains were susceptible to chloramphenicol, ampicillin and gentamicin. Two L. salivarius isolates and two L. reuteri isolates were found to be multiresistant. CONCLUSIONS: This study indicates that the faecal Lactobacillus flora in piglets consists mainly of L. reuteri, L. salivarius and L. acidophilus group lactobacilli, and the distribution of lactobacilli is similar between individuals of the same age and with the same diet. Most of the Lactobacillus isolates tested were sensitive to the antibiotics used in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: Valuable information on Lactobacillus species distribution and their antibiotic resistance profiles in piglets is obtained.