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G M Scalarone

Publications and source records attributed to G M Scalarone.

At least 19 recordsLinked to original sources
PubMed2005

Blastomyces dermatitidis antigen detection in urine specimens from dogs with blastomycosis using a competitive binding inhibition ELISA.

A competitive binding inhibition enzyme linked immunosorbent assay (ELISA) was used to detect Blastomyces dermatitidis antigens in urine specimens from dogs with blastomycosis. Sera from rabbits immunized with B. dermatitidis killed whole yeast cells were used as the primary antibody in the competitive ELISA. This initial study was performed to determine if B. dermatitidis antigen detection was possible and to test the efficacy of the rabbit sera as a primary antibody. An indirect ELISA was also performed to compare antigen detection in urine to antibody detection in the sera of the infected dogs. The results indicate 100% (36/36 specimens) detection of both antigen and antibody. Cross reactivity with Histoplasma capsulatum, as well as non-specific binding with the normal urine specimens, was observed with the competitive binding inhibition ELISA.

Animals↗
PubMed2004

Comparative serological evaluation of 10 Blastomyces dermatitidis yeast phase lysate antigens from different sources.

Yeast phase lysate antigens from 10 isolates of Blastomyces dermatitidis (dog, ERC-2 and T-58; T-27, polar bear; woodpile, ER-3; bat lung, 48938; human, B5929, B5895, B5896, B5931, and CAPP) from different geographical regions, in addition to a Histoplasma capsulatum (G217B) lysate preparation were compared with respect to their reactivity against serum specimens from dogs, rabbits and humans positive for blastomycosis using an indirect enzyme-linked immunosorbent assay. In addition, the lysate antigens were also assayed against histoplasmosis-positive human serum samples to study their cross-reactivity. Variable results were obtained with T-58 and T-27 exhibiting the greatest reactivity. We also noticed that the lysate did not react consistently to serum samples across species with lesser reactivity evidenced when testing dog sera. Finally, T-58 gave the highest cross-reactivity with histoplasmosis-positive sera. The study may prove valuable in the development of antigen candidates for blastomycosis serodiagnosis.

Animals↗
PubMed2003

Blastomyces dermatitidis lysate antigens: antibody detection in serial serum specimens from dogs with blastomycosis.

Yeast phase lysate antigens prepared from different isolates of Blastomyces dermatitidis (T-58, dog-Tennessee; T-27, polar bear-Tennessee; ERC-2, dog-Wisconsin; ER-3, woodpile-Wisconsin) were compared with respect to the detection of antibodies (indirect enzyme-linked immunosorbent assay-ELISA, peroxidase system) in 126 serial serum specimens (pre-treatment, 30 and 60 days post-treatment with itraconazole) from 42 dogs with diagnosed blastomycosis. Mean absorbance values observed with the four lysate antigens at the three treatment intervals ranged from the most reactive to the least reactive as follows: T-58 (0.270, 0.210, 0.136); T-27 (0.209, 0.156, 0.096); ER-3 (0.189, 0.144, 0.089) and ERC-2 (0.158, 0.129, 0.080). Even though variations in reactivity were evidenced, the lysates prepared from isolates from various geographical regions and sources were all efficacious as antigens for the immunodiagnosis of canine blastomycosis.

Animals↗
PubMed2002

Serological differences in two Blastomyces dermatitidis isolates from different geographical regions of North America.

Yeast phase lysate antigens were prepared from two isolates (T-58 and ERC-2) from different geographic locations. Tennessee and Wisconsin. These lysate were evaluated with respect to their ability to detect antibody in dogs infected with blastomycosis and rabbits immunized with the lysates by an enzyme linked immunosorbent assay (ELISA). Both the dog sera and rabbit sera assays demonstrated that there were serological differences in these two isolates, which implied that there was antigenic variance in geographical populations of B. dermatitidis. These results correlated with a previous molecular study that indicated that there are genetic differences in different geographical populations of the organism.

Animals↗
PubMed2002

Comparison of seven Blastomyces dermatitidis antigens for the detection of antibodies in humans with occupationally acquired blastomycosis.

Yeast phase lysate antigens, prepared from seven isolates of Blastomyces dermatitidis (dogs, T-58, M-98; human, Le; soil, S; sea lion, SL; polar bear, PB; cat, C) were assayed by enzyme-linked immunosorbent assay (ELISA) to detect antibodies in human sera. The sera were from individuals potentially exposed to B. dermatitidis while working on a prairie dog relocation project in Colorado. All antigens exhibited greatest reactivity with three specimens in two separate assays. Absorbance values ranged from 0.370 (M-98) to 0.427 (LE) (Trial 1) and from 0.579 (M-98) to 0.714 (PB) (Trial 2) with serum 2; from 0.368 (C) to 0.453 (LE) (Trial 1) and from 0.565 (SL) to 0.694 (S) (Trial 2) with serum 3 and from 0.392 (M-98) to 0.506 (LE) (Trial 1) and from 0.557 (SL) to 0.758 (T-58) (Trial 2) with serum 6. The greatest reactivity was observed with sera from two persons (2 and 3 from the same individual and serum 6) who became ill with symptoms of pulmonary disease and sought medical care. Both patients were subsequently diagnosed with blastomycosis by microscopic and culture methods. The study indicated that all of the lysate antigens, regardless of source, could be used to reliably diagnose blastomycosis.

Animals↗
PubMed2002

Serological differences in three Blastomyces dermatitidis strains.

Yeast phase lysate antigens, prepared from three isolates of Blastomyces dermatitidis (T-58, Tennessee dog; 48089, Zaire human; ERC-2, Wisconsin dog) were assayed for their ability to detect antibodies in human sera, dog sera and sera from rabbits immunized with each of the lysate antigens. The dog sera were from animals diagnosed with blastomycosis from various endemic regions in North America. T-58 and ERC-2 lysate antigens exhibited a high reactivity with the serum from dogs infected with blastomycosis; however, 48089 lysate showed low reactivity with the same sera. With the immunized rabbit sera, 48089 lysate was the only lysate with a high reactivity with the 48089 serum and it exhibited little reactivity with the heterologous sera. The T-58 and ERC-2 lysate antigens reacted minimally with the 48089 serum but reacted highly with both the T-58 and ERC-2 sera. The human sera were from individuals potentially exposed to B. dermatitidis while working on a prairie dog relocation project in Colorado. Remarkably, all three lysate antigens could detect antibodies in the individuals diagnosed with blastomycosis. This study indicated that there were serological differences in the 48089 Zaire lysate compared with the other lysate antigens and it may be designated serotype 2.

Animals↗
PubMed2001

Genetic diversity and transcriptional analysis of the bys1 gene from Blastomyces dermatitidis.

Blastomyces dermatitidis, a pathogenic fungal organism, is able to exist in two different morphologies, a multicellular mycelium or a unicellular yeast, according to temperature, 25 degrees C and 37 degrees C respectively. The switching between morphologies must be accompanied by a cascade of signaling events in which expression of genes responsible for the change of morphology is increased or decreased. bys1, a gene from B. dermatitidis isolate #58, is expressed at high levels in the unicellular yeast, but gradually diminishes as the temperature is lowered and the organism converts to the mycelial phase where there is no transcription of bys1. We explored if bys1 homologs are found in other B. dermatitidis isolates and if the transcription of the homologs were regulated by temperature. bys1 was identified in all B. dermatitidis isolates tested and could be grouped into two classes by Southern blot, PCR, and DNA sequence. Although the bys1 transcripts of both classes were regulated by temperature, transcription rates varied between the three isolates tested.

Amino Acid Sequence↗
PubMed1997

Immunological and chemical characterization of glycoproteins in IEF fractions of Blastomyces dermatitidis yeast lysate antigen.

After isoelectric focusing (IEF), fractions of a Blastomyces dermatitidis yeast lysate antigen were analysed for the presence of glycoproteins that may lead to cross-reactivity in immunoassays for the diagnosis of blastomycosis. Five major glycoproteins were apparent, two of which showed cross-reactivity when used in Western blots with sera obtained from dogs with histoplasmosis and coccidioidomycosis. These five glycoproteins were characterized for linkage to the proteins using N-glycosidase F (NGF) and for their lectin binding properties. The cross-reactive 235- and 160-kDa glycoproteins were found to possess mainly O-linked, high-mannose-type carbohydrates, and periodate-mediated oxidation of these molecules eliminated cross-reactivity observed with heterologous sera. Thus, the periodate-treated IEF antigens described here may be useful in solid-phase enzyme immunoassays for the diagnosis of blastomycosis.

Animals↗
PubMed1997

Comparative studies on the detection of antibodies and delayed hypersensitivity responses with 10 Blastomyces dermatitidis lysate antigens.

Yeast-phase lysate antigens were prepared from 10 different isolates of Blastomyces dermatitidis. Comparative studies were performed using the lysate antigens in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies in sera from dogs with blastomycosis and histoplasmosis. In order to evaluate the ability of the lysate reagents to elicit delayed dermal hypersensitivity (DTH) responses, the lysates were compared as skin-testing antigens in hairless guinea pigs that were previously sensitized with B. dermatitidis or Histoplasma capsulatum killed whole yeast cells. All ten of the lysate reagents were able to detect antibody with the ELISA in the serum specimens from dogs with blastomycosis (absorbance values ranged from 0.184 to 0.272; mean value 0.235). In contrast, when the lysates were assayed against sera from dogs with histoplasmosis, the absorbance values ranged from 0.053 to 0.151, with a mean value of 0.092. All ten lysate antigens were able to elicit a DTH response in the B. dermatitidis-immunized animals (mean axes of induration values ranged from 7.0 to 14.4 mm; mean value 8.6 mm). On the other hand, only minimal reactivity was evidenced in the guinea pigs immunized with H. capsulatum (mean axes of induration values ranged from 0.8 to 2.9 mm; mean value 1.8 mm).

Animals↗
PubMed1997

Detection of antibodies and delayed dermal hypersensitivity with different lots of Blastomyces dermatitidis yeast lysate antigen: stability and specificity evaluations.

Comparative evaluations were performed to assess the stability, sensitivity and specificity of eight lots of yeast lysate antigen prepared from a Blastomyces dermatitidis dog isolate (T-58). These antigens were prepared during the period from 1989 to 1995. The lysates were used in an ELISA for the detection of antibodies in serum specimens from dogs with blastomycosis and histoplasmosis. In order to evaluate the ability of the lysates to elicit delayed dermal hypersensitivity (DTH) responses, they were compared as skin-testing antigens in guinea pigs that were previously sensitized with B. dermatitidis or Histoplasma capsulatum killed whole yeast cells. All 8 of the lots of antigen detected antibody in the sera from dogs with blastomycosis (absorbance values ranged from 0.432 to 0.543; mean value of 0.508). The absorbance values ranged from 0.283 to 0.439 (mean value of 0.326) when the lysates were assayed against sera from dogs with histoplasmosis. All of the antigens were able to elicit a DTH response in B. dermatitidis immunized animals (mean axes of induration values ranged from 10.5 mm to 12.5 mm; mean value of 11.6 mm). In contrast, only minimal cross-reactivity was evidenced in the guinea pigs immunized with H. capsulatum (mean axes of induration values ranged from 0 to 4.5 mm; mean value of induration of 1.7 mm).

Animals↗
PubMed1992

Comparative studies on the postantifungal effect produced by the synergistic interaction of flucytosine and amphotericin B on Candida albicans.

A turbidometric method was used to measure Candida albicans yeast cell growth and to quantitate the postantifungal effect (PAFE) after exposure to various concentrations of flucytosine and amphotericin B, alone and in combination, for 2 hr at 30 degrees C. The drug concentrations used in the PAFE assays were determined by initial MIC and FIC (fractional inhibitory concentration) evaluations. The PAFE was calculated by the difference in time (hr) required for growth of the control and test cultures to reach the 0.5 absorbance level following removal of the drug by dilution. A synergistic PAFE was evidenced with combinations of the two drugs at concentrations below their individual MICs. Combinations of flucytosine (0.012 to 0.049 micrograms ml-1) and amphotericin B (0.195 to 0.39 micrograms ml-1) produced PAFEs ranging from 6.3 to 21.8 hr. These PAFEs persisted from 0.3 to 14.7 hr longer than those achieved when each of the two agents was assayed separately.

Amphotericin B↗
PubMed1992

Evaluation of a commercial DNA probe assay for the identification of clinical isolates of Blastomyces dermatitidis from dogs.

A DNA probe assay (AccuProbe, Gen-Probe, Inc., San Diego, CA) for culture confirmation of Blastomyces dermatitidis in clinical isolates was evaluated using 72 isolates of B. dermatitidis (62 dog isolates, five from other animals, four human isolates and one soil isolate). In addition, specificity determinations were performed using 28 American Type Culture Collection (ATCC) fungal isolates, other than B. dermatitidis, representing 21 different genera. The non-isotopic hybrid protection assay, that can be performed in less than 1 h, uses a chemiluminescent acridinium ester DNA probe that is homologous to ribosomal RNA sequences of B. dermatitidis. Positive results were obtained with all 72 B. dermatitidis isolates (100% sensitivity). In addition, no positive results were evidenced with the other 28 non-B. dermatitidis fungal isolates, including six strains of Histoplasma capsulatum (100% specificity).

Animals↗
PubMed1992

Synergistic postantifungal effect of flucytosine and fluconazole on Candida albicans.

The in vitro efficacy of flucytosine and fluconazole, separately and in combination, with respect to induction of a postantifungal effect (PAFE) on Candida albicans was studied. PAFE refers to the persistent suppression of fungal cell growth following a short period of exposure to an antifungal agent. A turbidometric method was used to measure cell growth and to quantitate the PAFE following exposure of C. albicans yeast cells to different concentrations of the two agents for 2 h. The PAFE was determined by the difference in time (h) required for growth of the control and test cultures to increase to the 0.5 absorbance level following removal of the drug by dilution. Minimum (MIC) and fractional inhibitory concentration determinations were made and the data used for selecting the concentrations used in the PAFE evaluations. A synergistic interaction of the two drugs at concentrations well below their individual MICs was evidenced. Flucytosine:fluconazole ratios of 1:16-1:32 at concentrations ranging from 0.024-0.098 micrograms ml-1 and from 0.78-1.56 micrograms ml-1, with flucytosine and fluconazole, respectively, induced PAFEs which persisted for 2.5 h longer than those achieved when each of the two agents was assayed separately.

Candida albicans↗
PubMed1992

The postantifungal effect of 5-fluorocytosine on Candida albicans.

The in-vitro postantifungal effect (PAFE) of 5-fluorocytosine for Candida albicans for short periods of time was investigated. Yeast cells were exposed for 0.5, 1 or 2 h to a range of concentrations (0.1-3.2 mg/L) of 5-fluorocytosine. The PAFE was quantitated by determinations of the number of colony forming units at hourly intervals (0-10 h) after removal of the drug by dilution. The length of the PAFE was dependent upon the concentration of 5-fluorocytosine and the duration of exposure. An exposure time of 0.5 h resulted in PAFE's ranging from 0 to 4.2 h. Exposure times of 1 and 2 h resulted in longer PAFEs and in many instances suppression of cell growth was seen for the entire evaluation period (up to ten hours).

Candida albicans↗
PubMed1991

Detection of antibody responses and delayed dermal hypersensitivity with Blastomyces dermatitidis yeast and mycelial lysate antigens.

Yeast cell lysate and mycelial lysate antigens prepared from one strain (T-58) of Blastomyces dermatitidis were evaluated with respect to the detection of antibodies and delayed dermal hypersensitivity. Comparable ELISA sensitivity values were evidenced with the two antigens when assayed against serum specimens from dogs with blastomycosis, sera from non-infected dogs residing in endemic and non-endemic areas for blastomycosis and sera from rabbits that were hyperimmunized with B. dermatitidis antigens. Specificity determinations with anti-Histoplasma capsulatum rabbit sera indicated that both reagents exhibited only minimal cross-reactivity; the mycelial antigen was slightly more specific than the yeast phase reagent. Similar sensitivity and specificity results were experienced when the two antigens were used to detect delayed dermal hypersensitivity in guinea pigs previously sensitized with B. dermatitidis or H. capsulatum.

Animals↗
PubMed1988

Preparation and ELISA evaluation of Blastomyces dermatitidis yeast phase lysate antigens.

Enzyme-linked immunosorbent assay (ELISA) comparative studies are described in which 10 Blastomyces dermatitidis antigenic preparations from human, dog, and soil strains were used for the detection of rabbit serum antibodies directed against B. dermatitidis killed yeast cells of human and dog origin as well as detection of antibodies in human serum samples. Reactive reagents were produced from all 10 strains, but two antigens (K-Le and T-043324) demonstrated exceptional sensitivity with moderate cross-reactivity with histoplasmosis specimens. Additionally, these reagents were able to detect anti-B. dermatitidis antibodies produced in rabbits in response to human (H-100) or dog (T-042976) strains in a comparable manner. These data suggest that a single antigenic reagent might be suitable for ELISA detection of B. dermatitidis antibodies in different species with blastomycosis.

Animals↗