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Biomedical subjects

W P Johnson

Publications and source records attributed to W P Johnson.

At least 19 recordsLinked to original sources
PubMed2001

Ferrographic tracking of bacterial transport in the field at the narrow channel focus area, Oyster, VA.

The first results from an innovative bacterial tracking technique, ferrographic capture, applied to bacterial transport in groundwater are reported in this paper. Ferrographic capture was used to analyze samples during an October 1999 bacterial injection experiment at the Narrow Channel focus area of the South Oyster site, VA. Data obtained using this method showed that the timing of bacterial breakthrough was controlled by physical (hydraulic conductivity) heterogeneity in the vertical dimension as opposed to variation in sedimentsurface or aqueous chemical properties. Ferrographic tracking yielded results that compared well with results from other tracking techniques over a concentration range of 8 orders of magnitude and provided a low detection limit relative to most other bacterial tracking techniques. The low quantitation limit of this method (approximately 20 cells/mL) allowed observation of transport of an adhesion-deficient bacterium over distances greater than 20 m in the fine sand aquifer underlying this site.

Bacteria↗
PubMed2001

Comparison of methods for monitoring bacterial transport in the subsurface.

The purpose of this study was to compare in a laboratory experiment, a suite of methods developed to track viable bacteria during field transport experiments. The criteria for development and selection of these methods included: (1) the ability to track bacteria within the environment from which they were isolated; (2) the lack of any effect upon the viability or the transport characteristics of the strain; (3) low detection limits; (4) a quantification range that covered several orders of magnitude; and (5) an analytical cost and turnover time commensurate with the analysis of several thousands of samples in a few months. The approaches developed included: enumeration of bacteria labeled with a vital fluorescent stain (CFDA/SE) using microplate spectrofluorometry, flow cytometry, and ferrographic (immunomagnetic) capture; enumeration of highly (13)C-enriched bacteria using combustion-IRMS; and quantitative PCR. These methods were compared to direct microscopic enumeration and plate counts during a bacterial transport experiment performed in an intact sediment core and designed to simulate the field experiment. Four of the seven methods had equivalent recoveries for the breakthrough of a pulse of bacteria eluting from a 50-cm long sediment core, and all of the methods detected the arrival of cells in the effluent prior to the conservative tracer. Combustion IRMS and ferrographic enumeration had the lowest quantification limits (approximately 2 to 20 cells/ml), whereas microplate spectrofluorometry had the highest quantification limit (approximately 10(5) cells/ml). These methods have the potential for numerous applications beyond tracking bacteria injected into the subsurface.

Bacteria↗
PubMed2001

Evidence for detachment of indigenous bacteria from aquifer sediment in response to arrival of injected bacteria.

Two bacterial strains isolated from the aquifer underlying Oyster, Va., were recently injected into the aquifer and monitored using ferrographic capture, a high-resolution immunomagnetic technique. Injected cells were enumerated on the basis of a vital fluorescence stain, whereas total cell numbers (stained target cells plus unstained target and antigenically similar indigenous bacteria) were identified by cell outlines emanating from fluorophore-conjugated antibodies to the two target strains. The arrival of injected bacteria at the majority of monitored sampling ports was accompanied by simultaneous temporary increases in unstained cell counts that outnumbered the injected bacteria by 2- to 100-fold. The origin and mechanism of appearance of the unstained cells are considered.

Bacterial Adhesion↗
PubMed1995

An integrated strategy for structural characterization of the protein and carbohydrate components of monoclonal antibodies: application to anti-respiratory syncytial virus MAb.

The relatively rapid and extensive characterization of the amino acid sequence and site-specific carbohydrate structures of a recombinant, reshaped human monoclonal antibody directed against respiratory syncytial virus (RSHZ19) is presented. The integrated strategy used a combination of mass spectrometric and conventional methodologies. Liquid chromatography/electrospray mass spectrometry was used for peptide mapping and selective identification of glycopeptides, and Edman degradation and tandem mass spectrometry were used to define the sequences of selected peptides. Matrix-assisted laser desorption/ionization mass spectrometry provided the M(r) of the intact protein and was used to characterize endo- and exoglycosidase digests of isolated glycopeptides to identify the glycosylation-site peptide and define the structures of the carbohydrates at that site. These experiments verified 99.1% of the light- and 99.3% of the heavy-chain amino acid sequences. The N and C termini of both chains were confirmed, and the nature and extent of heterogeneity at the N and C termini of the heavy chain were determined. Oxidation of a specific methionine residue to the sulfoxide was demonstrated by sequencing the N-terminally blocked peptide by tandem MS. Carbohydrate was found exclusively at Asn296 of the heavy chain. There was no evidence for a nonglycosylated form of the molecule or for the presence of O-linked carbohydrate. The qualitative distribution of glycoforms at this site was determined by MS of the isolated, tryptic glycopeptide and compared with results obtained by high-performance anion exchange chromatography and high-resolution gel permeation chromatography of oligosaccharides released by hydrazinolysis. The sequence and linkage of individual glycan species were determined using matrix-assisted laser desorption/ionization MS to monitor the results of a series of controlled digestions with specific exoglycosidases. The set of glycoforms consists predominantly of biantennary, core fucosylated carbohydrates lacking sialic acid. The present study is one of the first to directly evaluate the quantitative as well as qualitative consistency of the MS methods with conventional methods for carbohydrate analysis.

Amino Acid Sequence↗
PubMed1976

Sororal occurrence of diffuse esophageal spasm.

In this report, we have described two sisters with classical symptoms, radiological appearances, and manometric features of diffuse esophageal spasm. To the best of our knowledge, familial occurrence of this disorder has not been documented previously.

Esophageal Diseases↗
PubMed1976

Responses of the competent and incompetent lower oesophageal sphincter to pentagastrin and abdominal compression.

Responses of the lower oesophageal sphincter (LOS) to pentagastrin, given by continuous intravenous infusion in doses ranging between 0 and 9 mug/kg/h, and to external abdominal compression were measured by infused catheters in healthy subjects and in a group of patients with reflux oesophagitis. In separate experiments, pressures were measured both by sensors stationary within the LOS, and by repeated continuous withdrawals of sensors from stomach to distal oesophagus. In normal subjects, doses of pentagastrin within the physiological range (0.9 mug/kg/h) produced modest but statistically significant increases in LOS pressure. By comparison, sphincteric responses in patients with oesophagitis were small and a dose of 4-5 mug/kg/h was the lowest that produced a significant increase in LOS pressure. During abdominal compression increases in LOS pressure did not significantly exceed increases in intragastric pressure in either patients or normal subjects. This was so at all doses of pentagastrin that were tested. Hence, there was no evidence of synergism between the effects of pentagastrin and abdominal compression upon the LOS. We infer from our findings that gastrin does play a modest role in the physiological regulation of human LOS tone. Relative insensitivity of the incompetent LOS to pentagastrin represents, we believe, sphincteric muscle failure. Our results are not consistent with the hypothesis that LOS incompetence is due to loss or impairment of an adaptive response of the LOS to alterations in intra-abdominal pressure.

Abdomen↗
PubMed1972

In vivo formation of H2O2 in red cells during exposure to hyperoxia.

Chow-fed and tocopherol-deficient mice were given aminotriazole (AT), exposed to 100% O(2) at 60 pounds per square inch absolute for 1 hr (OHP), and red blood cells were assayed for catalase activity and lipid peroxide levels. A decrease of catalase activity (CA) in the presence of AT can be taken as evidence of excess formation or accumulation of H(2)O(2). No differences of CA were observed among chow-fed mice, with or without AT and/or OHP. Tocopherol-deficient mice with AT had lower CA (0.174+/-0.040) than chow-fed mice with AT (0.225+/-0.028) P < 0.01. Tocopherol-deficient mice with AT exposed to OHP had even lower CA, 0.137+/-0.024, P < 0.01.The data are consistent with the hypothesis that H(2)O(2) is formed or accumulated in excess in red cells of tocopherol-deficient mice, an effect that is enhanced in the presence of hyperoxia. They imply that tocopherol plays a role in the detoxification of H(2)O(2).

Animals↗