Visualizing adhesion-induced agglutination of Escherichia coli with mannosylated nanoparticles.

Polymeric nanoparticles covalently functionalized with derivatized D-mannose molecules were synthesized and characterized. These nanoparticles have an average size of approximately 160 nm in diameter, thus bearing a large number of surface-tethered mannose moieties for multivalent interactions with adhesins on bacterial cells. Specifically, the mannosylated nanoparticles bind strongly with Escherichia coli, allowing the convenient visualization of adhesion interactions under a conventional electron microscope.

Use of a Disposable Syringe as a Novel Economic Breath-Collection Chamber for Mice.

We modified commercially available 50-ml syringes to allow their use as breath-collection chambers for mice. By drilling holes at the flanged end and applying 3-way valves to the opposite (tip) end, a syringe can be ventilated, then quickly and simply converted to a closed chamber for collection of breath samples. We evaluated the influence of sample collection time on CO2 content of breath samples and found that 30 sec was required to obtain a CO2 concentration of 3 to 5%.

Vector potential of houseflies for the bacterium Aeromonas caviae.

Houseflies, Musca domestica Linnaeus (Diptera: Muscidae), have been implicated as vectors or transporters of numerous gastrointestinal pathogens encountered during feeding and ovipositing on faeces. The putative enteropathogen Aeromonas caviae (Proteobacteria: Aeromonadaceae) may be present in faeces of humans and livestock. Recently A.

Detection of Aeromonas caviae in the common housefly Musca domestica by culture and polymerase chain reaction.

Aeromonas caviae has been implicated in diarrhoeal disease of livestock and humans. The potential role of houseflies in the epidemiology of this pathogen was investigated by examining the prevalence of A. caviae in houseflies collected from two South Carolina farms and one restaurant.

Factors affecting the validity of the 13C-urea breath test for in vivo determination of Helicobacter pylori infection status in a mouse model.

Background: The mouse model using a human isolate of Helicobacter pylori is being widely accepted as an economical means of studying gastric infection. A noninvasive monitoring method would be useful for repeated testing to establish the time course of infection and the efficacy of treatments. In this study, we describe factors that affected interpretation of 13C urea breath test results for the assessment of H.

Repression and inactivation of α-amylase in species during growth on cellobiose.

Thermophilic actinomycetes establish themselves as numerically dominant bacterial populations in selected high temperature environments by virtue of their exoenzymic ability to degrade the complex polysaccharides in thermogenic plant biomass. When were grown on a mixture of cellulose and starch in mineral salts minimal medium, α-amylase was repressed via inhibition of maltose uptake by cellobiose. Addition of cellobiose to exponential phase cells growing on maltose or maltotriose triggered rapid degradation of extant amylase in the culture fluid of wild-type cells, but not in a protease-deficient mutant of A serine protease purified from caused inactivation of the amylase in culture fluid of the mutant when added at a concentration approximating to that of the wild-type strain.

Effect of carbon source on growth temperature and fatty-acid composition in Thermomonospora curvata.

The growth-temperature range of the actinomycete, Thermomonospora curvata, was influenced by the nature of the soluble carbon sources used, which were derived from cellulose, pectin, starch and xylan. This thermophile had the broadest (38 to 65°C) and narrowest (42 to 59°C) temperature range during growth on cellobiose (from cellulose) and 4-deoxy-LXXX-threo-t-hexoseulose uronic acid (from pectin), respectively. This substrate-temperature interaction was accompanied by changes in cellular fatty acids: uronic-acid-grown cells had relatively low amounts of branched chain fatty acids (particularly iso-16:0) and high amounts of monounsaturated fatty acids (particularly cis-18:1) compared with cells grown on any other substrate.

Purification and characterization of the major beta-1,4-endoglucanase from Thermomonospora curvata.

The major beta-1,4-endoglucanase (EG) of the thermophilic actinomycete, Thermomonospora curvata, contributed over 80% of the total EG activity recovered from cell-free culture fluid after growth on cellulose. The enzyme was purified to electrophoretic homogeneity by ammonium sulphate precipitation, ion-exchange chromatography and size exclusion HPLC. This monomeric enzyme had a specific activity of 750 IU mg(-1) when assayed with 2.

Adaptation of the [13C]urea breath test as a noninvasive method for detection of Helicobacter pylori infection in squirrel monkeys (Saimiri spp.).

The [13C]urea breath test was adapted for use in squirrel monkeys (Saimiri spp.) for identification of experimentally induced infection with Helicobacter pylori, the bacterium causing gastric ulcer in humans. A canine anesthesia inhalation mask was modified with a volume-reducing insert allowing sufficient breath collection from these small primates within 30 sec.

Effects of the detergent Tween 80 on Thermomonospora curvata.

Tween 80 (0.1%, v/v) added to Thermomonospora curvata growing in minimal medium caused a transient lowering of the dry cell mass, decreased the optimal growth temperature of the thermophile from 62 to 54°C, and increased extracellular esterase activity. Cells grown in the presence of Tween 80 had decreased concentrations of branched chain fatty acids and increased concentrations of oleic acid.

Interference of the detergent Tween 80 in protein assays.

The nonionic detergent Tween 80, which has been widely used to stimulate protein secretion in bacterial and fungal systems, caused interferences in three protein determination methods. The OD595 developed in the Coomassie blue dye-binding assay with a variety of purified proteins in the presence of Tween 80 was 1.6 to 3.

Influence of carbon source on cell surface topology of Thermomonospora curvata.

The appearance of cell surface protuberances in Thermomonospora curvata correlated with cell-bound exoenzymes which could be removed by brief sonication. Mycelia grown on cellulose or xylan had numerous protuberances and retained 20 to 25% of endoglucanase and endoxylanase at cell surfaces, while those grown on pectin or starch had few protuberances and negligible bound pectinase or amylase.

Temperature-dependent patterns of exoenzyme biosynthesis in Thermomonospora curvata.

Production of depolymerlzing exoenzymes in the thermophilic actinomycete, Thermomonospora curvata, grown at 40°, 50° and 61°C, were compared. Cellulase-specific activities were similar at the three growth temperatures. Amylase-and pectinase-specific activities decreased with increasing growth temperature, while xylanase had the reverse pattern.

Cloning of three endoglucanase genes from Thermomonospora curvata into Escherichia coli.

A BamHI genomic library from Thermomonospora curvata was constructed in E. coli using cosmid vector pHC79. Four clones able to hydrolyze CMC were isolated.

Changes in Endoglucanase Patterns during Growth of Thermomonospora curvata on Cellulose.

The endoglucanases of the thermophilic actinomycete Thermomonospora curvata were characterized. Early-exponential-phase culture fluid contained at least three endoglucanases, with molecular weights of 23,000, 46,000, and 146,000 and K(m) values of 1.54, 3.

Preferential Utilization of Cellobiose by Thermomonospora curvata.

Thermomonospora curvata was cultivated on mineral salts medium containing glucose and cellobiose under conditions that increasingly favored the uptake of glucose. In each case cellobiose was utilized in preference to glucose and induced beta-glucosidase and endoglucanase activity. [C]glucose metabolism studies indicated that cellobiose was not cleaved by extracellular beta-glucosidase and transported as glucose.

Inducible thermoalkalophilic polygalacturonate lyase from Thermomonospora fusca.

A thermostable polygalacturonate lyase (PL; EC 4.2.2.

Cyclic AMP phosphodiesterase in Thermomonospora curvata.

Cyclic AMP phosphodiesterase (PDE; EC 3.1.4.

Cyclic AMP levels during induction and repression of cellulase biosynthesis in Thermomonospora curvata.

Specific cellulase production rates (SCPR) were compared with intracellular cyclic AMP (cAMP) levels in the thermophilic actinomycete, Thermomonospora curvata, during growth on several carbon sources in a chemically defined medium. SCPR and cAMP levels were 0.03 U (endoglucanase [EG] units) and 2 pmol per mg of dry cells, respectively, during exponential growth on glucose.

Cellulase biosynthesis in a catabolite repression-resistant mutant of Thermomonospora curvata.

A catabolite repression-resistant mutant of the thermophilic actinomycete Thermomonospora curvata was obtained by treatment with ethyl methanesulfonate and UV light. Cellulase biosynthesis was undiminished by glucose, 2-deoxyglucose, or alpha-methyl glucoside, which are potent repressors in the wild type. Intracellular cyclic AMP levels were higher in the mutant in both the absence and the presence of repressors.

Quantitative screening of clinical isolates for immunoglobulin A protease production.

The production of immunoglobulin A (IgA) protease is a potentially useful marker in differentiating pathogenic from nonpathogenic species of clinical isolates; however, current quantitative assay methods are too tedious for routine application. A simple quantitative method was developed to screen clinical isolates for IgA protease production. This method is based on the specificity of reaction between IgA and alpha chain-specific antiserum in an immunochemistry analyzer (Beckman Instruments, Inc.