Charles D. Jeffries

Analytical isoelectric focusing of secreted dermatophyte proteins applied to taxonomic differentiation of Microporum and Trichophyton species (preliminary studies)

Culture filtrates were prepared from dermatophytes under standard conditions and adapted for analytical isoelectric focusing in thin layer polyacrylamide gels over the pH range 3.5-9.5. Dermatophytes grown in trypticase soy broth secreted a large number of proteins displaying a wide range of isoelectric points (pIs). Trichophyton megninii extracts contained a triplet of proteins focusing in the pH 8.0-8.5 range that were absent in taxonomically related T. kuryangei isolates. Single ascospore isolates and standard tester strains of Nannizzia otae (+) mating type were differentiated from the (-) mating type by proteins focusing at pH 6.5 and 8.4. These were markedly reduced in the (+) type. The isofocused pattern of Microsporum canis conformed closely to the (-) mating type of N. otae. The protein patterns of T. megninii and T. kuryangei were distinct from those obtained with M. canis and M. equinum because of an intense-staining broad protein band, pI 7.2, and three periodic acid-Schiff-positive glycoproteins focusing in the acidic range which were absent in the Microsporum species. A characteristic protein or doublet (pI 8.7) was present in the Microsporum species and absent in the Trichophyton species. Analytical isoelectric focusing is a potentially useful method to distinguish inter- and intra-species differences in the pattern of secreted dermatophyte proteins present in culture filtrates and in trichophytins. The information derived may be useful in the classification of species.

Curvularia lunata in experimental phaeohyphomycosis

Curvularia lunata strain SP, isolated from a disseminated human infection, infected normal mice, but three other strains of C. lunata and one each of C. pallescens and C. spicifera did not. The SP strain was recovered in cultures from, and hyphal filaments were observed in, abscesses in the liver and spleen of experimentally infected mice. All strains of Curvularia infected mice treated with 400 rads X-irradiation and 10.0 mg cortisone, but at 400 rads and 5.0 mg only two strains of C. lunata (SP and Ghosh) and C. pallescens and C. spicifera infected mice. At 200 rads and 10.0 mg, C. lunata Sp, C. pallescens and C. spicifera; and at 200 rads and 5.0 mg, only C. lunata SP and C. pallescens caused infection. After X-irradiation (200 or 400 rads) or cortisone (5.0 or 10.0 mg) alone only C. lunata SP caused infections in mice.

Class-specific antibody in human dermatophytosis reactive with Trichophyton rubrum derived antigen.

Dermatophyte infections induce a humoral immune response and an enzyme linked immunosorbent assay was used to detect specific antibody classes against antigen derived fromTrichophyton rubrum. Sera from 19 acute patients, 18 chronic patients, and 27 normal controls were evaluated. Mean IgG titers against dermatophyte antigen were significantly higher in all patients than in controls. Mean IgM levels were significantly higher in acute patients than in controls. No significant difference was detected in IgE titers between the patients and controls. These results do not reveal whether the humoral immune response has a role in the progression of the infection.

The Effect of Glutathione on Survival of Endotoxin Treated Mice

Conclusion Injection of mice with 30 mg of GSH, especially intravenously, was complicated by the unphysiologic, extracellular distribution of the exogenous glutathione. The GSH given to mice to replace nonprotein sulfhydryl lost as a result of endotoxin shock provided only slight protection. Survival was significantly improved by intraperitoneal injection of 30 mg of GSH 4 hr before the endotoxin. Thus, there appears to be little therapeutic potential for GSH in endotoxin shock.

Opportunistic fungi in detroit's rats and opossum

Four Norway rats (Rattus norvegicus) and one opossum (Didelphis virginiana) were surveyed for the presence of fungi. Eighteen different fungi were isolated from rat samples and six from the opossum, includingTrichophyton mentagrophytes. Hair and skin scrapings from laboratory raised Norway rats, bedding and air from the room yielded onlyPenicillium sp.

Effect of exogenous steroids and inhibitors of steroidogenesis on endotoxin shock.

Summary Suppression of corticosteroid production by inhibitors effected a tenfold increase in susceptibility of mice to death from endotoxin shock. Amphenone B and metyrapone caused some increase in susceptibility, but if given more than 1 hr before the endotoxin afforded protection to the test animals. Aminoglutethimide exerted a deleterious effect for at least 24 hr. The effect of the aminoglutethimide was reversed by pregnenolone, progesterone and deoxycorticosterone, although the LD50 remained lower than in the control animals. Corticosterone, cortisone and hydrocortisone afforded marked protection in both the endotoxin treated mice as well as the endotoxin-inhibitor treated mice. Andros-tenedione caused a decrease in the LD50 value in both the control and aminoglutethimidetreated mice. By contrast 17-a-hydroxyprogesterone afforded no protection against endotoxin in the control mice and a significant, but miniscule, degree of protection in the inhibitor-treated mice. This investigation was supported by a U.S. Public Health Service research grant AI-06767, from the National Institute of Allergy and Infectious Diseases.

Liver Carbohydrate Levels in Mice Treated with Endotoxin, Cortisone, and Elipten

Summary Mice treated with endotoxin 18 hr earlier had markedly reduced liver carbohydrate, but with cortisone under the same conditions the level was more than doubled. Elipten exerted no apparent effect on the carbohydrate levels in normal mice, but very nearly eliminated the deposition of glycogen after the mice were treated with cortisone. The effect of endotoxemia on the liver carbohydrate was enhanced by Elipten, and ameliorated slightly by cortisone alone or in combination with Elipten. The author thanks Dr. J. J. Chart of CIBA for the supply of Elipten. This investigation was supported by USPHS research grant AI-06767 from the National Institute of Allergy and Infectious Diseases.

Determination of Microbial DNA Base Pair Ratios by Agar Gel Electrophoresis

Summary The determination of DNA base pair ratios has been described using a rapid agar-gel electrophoresis method for the separation of DNA bases obtained from formic acid hydrolyzates. The actual mol % G + C values calculated from spectrophotometric data on the electrophoretically separated bases compared favorably with values obtained by thermal denaturation or buoyant density determinations on separate aliquots of the same DNA.