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Biomedical subjects

L D Gray

Publications and source records attributed to L D Gray.

32 records · Page 2Linked to original sources
PubMed1988

Laboratory diagnosis of systemic fungal diseases.

The increase in the number of fungal infections seen in debilitated and immunocompromised patients in the last several years makes it necessary to consider all fungi as potential pathogens. Clinical microbiology laboratories are playing increasingly important roles in the recovery, isolation, and identification of these fungi. This article contains specific recommendations and references concerning a practical approach to the laboratory identification of systemic fungi. The proper and timely selection, collection, and transport of specimens is imperative, and clinicians are responsible for appropriate specimen selection to ensure optimal chances of recovery of pathogens. Respiratory tract secretions and blood are excellent sources for detection of disseminated fungal infection. Specimens should be placed into transport media if the sample size is small or if only a small number of organisms are thought to be present. Direct microscopic examination of specimens can provide valuable information, often allowing a clinician to initiate immediate therapy. Specimens that are more likely than others to contain systemic fungi and that should be examined routinely include the following: pulmonary biopsy material, bronchial washes and lavages, specimens from immunocompromised patients, purulent specimens, and specimens suspected of containing a specific fungus. Valuable methods of examining specimens directly include treatment with KOH and calcofluor white. Use of media to recover fungi is the basis of making a laboratory diagnosis of a fungal disease, and the use of proper recovery and subculture media is imperative. Noninhibitory media allow contaminants to grow readily and should be used only to recover fungi from normally sterile body sites or for subculture. Blood-enriched media allow almost all pathogenic and saprophytic fungi to flourish. Therefore, such media, unless they contain antibiotics, should not be used as primary recovery media. Media that contain antibiotics should be used as primary recovery media to prevent overgrowth of pathogenic fungi by contaminants. Yeasts recovered from clinical specimens can be identified by a combination of tests, which include direct microscopic examination, germ tube formation, microscopic morphology of growth on corn meal agar, and ability to utilize certain carbohydrates. Molds recovered from clinical specimens are identified by a combination of growth rate, colonial characteristics, size and shape of hyphae, and microscopic examination of reproductive structures and other fungal elements.(ABSTRACT TRUNCATED AT 400 WORDS)

Culture Media↗
PubMed1988

Use of a semi-automatic image analyser in directly determining mean linear intercepts.

A direct method for measuring mean linear intercepts (Lm) was tested. A semi-automatic image analysis system (Zeiss, Videoplan) and an overlay of parallel lines were employed in determining Lm in intravascularly perfused guinea pig lungs. Actual chord lengths were recorded and the data was immediately processed. Spurious intercepts and manual data manipulation were avoided. This direct method of measurement proved to be simple, rapid and very consistent across three trials for each animal. Analysis of variance revealed no significant differences between the three trials (p = 0.22).

Animals↗
PubMed1987

Mouse skin damage caused by cytolysin from Vibrio vulnificus and by V. vulnificus infection.

Light and electron microscopy of mouse skin damage caused by intradermal infection with a virulent strain of Vibrio vulnificus and by a single intradermal injection of the cytolytic toxin produced by the bacterium revealed similar structural alterations. The epidermis was intact; however, the infection and toxin produced acute cellulitis characterized by extensive extracellular edema; disorganization of collagen bundles; large accumulations of cell debris and plasma proteins; damaged or necrotic fat cells, capillary endothelial cells, and muscle cells; and mild inflammatory cell infiltration. The virulent strain of V. vulnificus produced a capsule and was resistant to phagocytosis in vivo, whereas a weakly virulent strain of the bacterium did not produce a capsule and was readily phagocytized and digested. Factors that may be important in the pathogenesis of V. vulnificus wound infections include a capsule that inhibits phagocytosis and an extracellular cytolytic toxin that is responsible, at least in part, for the severe tissue damage characteristic of such wound infections.

Animals↗
PubMed1987

Effect of Candida morphology on amphotericin B susceptibility.

We showed that brief exposures to amphotericin B (AmB) inhibited the induction of new Candida germ tubes and the lengthening of partially induced germ tubes. Blastoconidia with germ tubes were more susceptible to AmB killing, and this varied directly with the induction period and the AmB exposure period. AmB did not preferentially affect germ tube adherence to fibrin matrices.

Amphotericin B↗
PubMed1987

Pulmonary lymphatic clearance of 99mTc-DTPA from air spaces during lung inflation and lung injury.

A total of 22 sheep with lymphatic cannulas were used to determine if 99mTc-labeled diethylenetriaminepentaacetic acid (DTPA) clears directly from the air spaces of the lungs into the lymph vessels. Each sheep was anesthetized and ventilated with an aerosol of the DTPA for 2-5 min, and the DTPA activities in the lymph and plasma were measured every 15 min for 2 h. After the first 45 min, the average ratio of the DTPA in the lymph to that in the plasma (L/P) was 1.03 +/- 0.06 (SD) in the six control experiments and 1.11 +/- 0.05 in the six experiments in which the lungs were inflated with a positive end-expired pressure of 10 cmH2O throughout the study. Direct movement of the DTPA from the air spaces into the lymph was not necessary to account for the DTPA clearance in these experiments because the L/P ratio was not significantly different from 1.0. Eight additional sheep received intravenous infusions of air at 0.2 ml.kg-1.min-1 for 2 h to induce lung injury before depositing the DTPA. In these sheep L/P was 1.53 +/- 0.28, which was significantly higher than the value measured in the control group (P less than 0.01). We considered the possibility that the increased L/P ratio in these sheep could be due to alterations in the distribution of the blood flow to the tissue, but the L/P ratio in four sheep whose distribution of blood flow was altered by inflation of a balloon in the right pulmonary artery was 1.05 +/- 0.10, the same as the control value.(ABSTRACT TRUNCATED AT 250 WORDS)

Aerosols↗
PubMed1986

Detection of anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human surviving V. vulnificus disease.

An enzyme-linked immunosorbent assay and a cytolysin neutralization assay were used detect anti-Vibrio vulnificus cytolysin antibodies in sera from mice and a human that survived V. vulnificus disease. The detection of antibodies against the cytolysin indicated that the cytolysin is produced in vivo, and this observation is consistent with the hypothesis that the cytolysin is involved in the pathogenesis of V. vulnificus disease.

Animals↗
PubMed1985

Identification of Vibrio vulnificus by indirect immunofluorescence.

A rapid, sensitive, and specific indirect immunofluorescence (IIF) procedure is described for identifying Vibrio vulnificus. Reference antisera were prepared by vaccinating rabbits with surface antigen preparations of V. vulnificus, and the antisera were examined for the ability to react with and serologically group 85 isolates of V. vulnificus grown in heart infusion broth, and to detect V. vulnificus in tissue specimens from mice experimentally infected with a virulent isolate of the bacterium. The antisera detected 100% of the V. vulnificus isolates examined and gave false-positive results in approximately 0.9% of 445 IIF tests performed with non-V. vulnificus clinical isolates. V. vulnificus also was detected in frozen tissue sections from infected mice; however, the most easily observed positive results were obtained by examining V. vulnificus from lesion specimens and blood cultured briefly in heart infusion broth. The bacteria in 2-hr-old cultures of local lesions fluoresced brilliantly and were easily detectable. The IIF procedure could be of value in rapidly diagnosing fulminating and potentially fatal human disease caused by V. vulnificus.

Animals↗
PubMed1985

Purification and characterization of an extracellular cytolysin produced by Vibrio vulnificus.

An extracellular cytolytic toxin produced by the halophilic bacterium Vibrio vulnificus was isolated free of detectable contamination with medium constituents and other bacterial products by sequential ammonium sulfate precipitation, gel filtration with Sephadex G-75, hydrophobic interaction chromatography with phenyl-Sepharose CL-4B, and isoelectric focusing in an ethylene glycol density gradient. The cytolysin is a heat-labile, hydrophobic protein that is inhibited by large amounts of cholesterol, is partially inactivated by proteases and trypan blue, has a molecular weight (estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by amino acid analysis) of ca. 56,000, and has an isoelectric point of ca. 7.1. The first 10 amino-terminal amino acid residues of the cytolysin are Gln-Glu-Tyr-Val-Pro-Ile-Val-Glu-Lys-Pro. Lysis of mouse erythrocytes by the purified cytolysin is a multi-hit, at least two-step process consisting of a temperature-independent, toxin-binding step, followed by a temperature-dependent, membrane-perturbation step(s). In addition to possessing cytolytic activity against erythrocytes from 17 animal species and against Chinese hamster ovary cells in tissue culture, the purified cytolysin preparation was lethal for mice (ca. 3 micrograms/kg, intravenous 50% lethal dose) and had vascular permeability factor activity in guinea pig skin.

Amino Acids↗
PubMed1984

Protection of mice against Vibrio vulnificus disease by vaccination with surface antigen preparations and anti-surface antigen antisera.

Vaccination of mice with either Formalin-killed cells or cell extracts of a virulent strain and a weakly virulent strain of Vibrio vulnificus or with rabbit antisera (AS) against the Formalin-killed cells and cell extracts protected against the virulent strain of V. vulnificus. However, the virulent strain vaccines and AS elicited a significantly stronger immune response than the weakly virulent strain vaccines and AS. Adsorption of the AS with either the homologous or heterologous Formalin-killed cells significantly reduced the ability of the AS to protect mice. The major protective antigen(s) in the cell extracts migrated in the void volume of Sephacryl S-400 superfine, was not effectively sedimented by centrifugation at 100,000 X g for 2 h, had an isoelectric point of 3.8 to 4.2, and was sensitive to boiling or autoclaving for 15 min, periodate oxidation, and exposure to pH 12 but was resistant to 56 degrees C, trypsin, pronase, RNase, neuraminidase, and pH 4.5. Electron microscopy revealed that the virulent strain possessed a more dense ruthenium red-staining layer on its outer membrane and had a much smoother surface than the weakly virulent strain. Our results provide evidence that a major protective antigen and virulence factor of V. vulnificus is a heat-labile, acidic polysaccharide located on the bacterial surface.

Animals↗
PubMed1978

Purification of Pseudomonas aeruginosa proteases and microscopic characterization of pseudomonal protease-induced rabbit corneal damage.

Extracellular proteases of three cornea-virulent strains of Pseudomonas aeruginosa were isolated by sequential ammonium sulfate precipitation, Ultrogel AcA 54 gel filtration, and flat-bed isoelectric focusing. The purity of the preparations was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis , thin-layer isoelectric focusing in polyacrylamide gel, immunodiffusion and immunoelectrophoretic procedures, and tests for the presence of other known pseudomonal products. Light and electron microscopic examination of rabbit corneal lesions observed 4 to 6 h after the intracorneal injection of submicrogram amounts of the proteases revealed: (i) degeneration and necrosis of epithelium, endothelium, and keratocytes, (ii) infiltration, degeneration, and necrosis of polymorphonuclear leukocytes, (iii) loss of the characteristic weblike pattern, colloidal iron staining, and ruthenium red staining of the stromal proteoglycan ground substance, (iv) dispersal of strucutrally normal appearing collagen fibrils, ground substance, (iv) dispersal of structurally normal appearing collagen fibrils, and (v) accumulation of plasma proteins and fibrin in the necrotic corneas. These structural alterations are very similar to those observed previously during experimental P. aeruginosa keratitis, and this similarity supports the idea that pseudomonal proteases are responsible, at least in part, for the rapid and extensive liquefaction necrosis characteristic of pseudomonal-induced keratitis. In addition, the results support the idea that pseudomonal proteases elicit severe corneal damage by causing the loss of the corneal proteoglycan ground substance, thus resulting in dispersal of undamaged collagen fibrils, weakening of the corneal stroma, and subsequent descemetocele formation and corneal perforation by the anterior chamber pressure.

Animals↗
PubMed1975

Rabbit corneal damage produced by Pseudomonas aeruginosa infection.

Gross, light microscopic, and electron microscopic examination of the rabbit corneal destruction produced by experimental Pseudomonas aeruginosa infections revealed a combination of acute inflammation and liquefaction necrosis of the cornea. Degeneration of the epithelial cells and the start of polymorphonuclear leukocyte infiltration of the cornea occurred initially. These changes were followed by loss of the epithelium, degeneration and loss of the keratocytes and endothelium, loss of the characteristic weblike pattern of the proteoglycan ground substance, dispersal of ultrastructurally normal collagen fibrils, extensive accumulation followed by degeneration of polymorphonuclear leukocytes, and accumulation of plasma proteins and fibrin in the necrotic cornea. Histochemical examination of the cornea suggested a loss of the proteoglycan ground substance but not of collagen. Rabbit corneas injected with Clostridium histolyticum collagenase showed gross and cellular changes similar to those observed during the pseudomonal infections; however, histochemical examination suggested a loss of collagen, and electron microscopy revealed ultrastructurally abnormal collagen fibrils. The results support the idea (i) that a bacterial or host-derived collagenase is not required for extensive corneal damage during a P. aeruginosa corneal infection, and (ii) that a P. aeruginosa corneal infection may severly damage the cornea by producing extensive corneal edema and by causing the loss of the corneal proteoglycan ground substance, thus resulting in dispersal of undamaged collagen fibrils, weakening of the cornea, and subsequent descemetocele formation and corneal perforation by the anterior chamber pressure.

Clostridium↗
PubMedDate not supplied

Correlation of urine pH with the detection of cytomegalovirus by the shell vial technique.

The influence of the pH of urine on the detection of cytomegalovirus (CMV) by the shell vial assay was evaluated. The pH values of 295 urine specimens ranged from 4.7 to 8.5 (mean 6.3) and were not significantly different in culture-positive versus culture-negative samples. None of the urine specimens appeared to be toxic for the cells used in the shell vial assay. We recommend inoculation of urine specimens into shell vials without adjustment of pH.

Cytomegalovirus↗
PubMedDate not supplied

The incidence of Mycoplasma pneumoniae pneumonia.

BACKGROUND: Mycoplasma pneumoniae has been considered a pathogen for humans since the 1940s. Of the 12 species of Mycoplasma found in humans, M pneumoniae is the most widely recognized pathogen. Morbidity from M pneumoniae results from the combined direct effect of cytotoxins produced by the organisms and the indirect effect of inflammatory responses to the presence of the organisms. Several studies have reviewed the incidence of M pneumoniae pneumonia in selected populations with variable results. By using tests that were not definitive detectors of the organism, earlier studies cited have overestimated the true incidence of this infection. We reevaluate several of these early studies in the light of newer findings. METHODS: Using the key words "Mycoplasma pneumoniae," "pneumonia," "prevalence," "incidence," and "community acquired," the MEDLINE files from 1992 to the present were searched. Articles dating before 1992 were accessed from cross-reference of the more recent articles. Only clinical trials with a sample size greater than 125 were considered. RESULTS AND CONCLUSIONS: M pneumoniae pneumonia occurs in 4- to 5-year cycles and in densely populated areas. Clinical symptoms of M pneumoniae pneumonia include dry cough, sore throat, middle ear involvement, and low-grade fever, as well as additional extrapulmonary manifestations. Bullous myringitis is not a common finding in M pneumoniae infection. Diagnostic tests include cold agglutinins, complement fixation, culture, and enzyme immunoassay. A fourfold rise in M pneumoniae-specific antibody in serum from acutely ill and convalescent patients remains the reference standard for diagnosing the infection. The incidence of M pneumoniae is probably lower than reported in many studies. Using tests that are not diagnostic of the infection can give a falsely elevated incidence of M pneumoniae infection in specific populations.

Humans↗