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S F Queener

Publications and source records attributed to S F Queener.

119 records · Page 7Linked to original sources
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Incorporation of fatty acids and amino acids by cultured Pneumocystis carinii.

Cultured P. carinii rapidly took up a variety of fatty acids. The relative rates of uptake for four fatty acids were 18:1 >> 16:0 approximately equal to 18:0 approximately equal to 18:2. Fatty acids were primarily incorporated into phospholipids and the uptake process was specifically inhibited by 2.2 and 22 microM primaquine, a concentration having no effect on host cells. Amino acids were also taken up by cultured P. carinii in a primaquine sensitive process. Radiolabeled leucine was incorporated into the major surface glycoprotein of P. carinii. The formation of radioactive P. carinii-specific proteins indicated that the cultured organisms carried out transcription and translation and that the incorporation of amino acids was dependent upon P. carinii rather than rare HEL human embryonic lung cells. The spinner flask culture system provides convenient access to P. carinii for metabolic studies in defined medium for a period of 5-14 days after inoculation.

Animals↗
PubMedDate not supplied

Isolation and characterization of rat lung Pneumocystis carinii gp120.

The principal glycoprotein (gp120) of Pneumocystis carinii obtained from infected rat lung was isolated by differential extraction and size-exclusion chromatography. The purified glycoprotein was cleaved with CNBr to two peptides of approximately 27 and 33 kDa. Amino acid sequences were obtained from both peptides. Proteolytic digestion with V8 protease yielded several peptides and sequences were obtained from peptides of 10 and 19 kDa. The cyanogen bromide cleavage results led to the conclusion that gp120 exists as a homodimer.

Amino Acid Sequence↗
PubMedDate not supplied

Inoculated mouse model of Pneumocystis carinii pneumonia.

A transtracheally inoculated mouse model of Pneumocystis carinii has been developed using BALB/c mice, a widely available strain free of latent P. carinii infection. The mean infectivity score of untreated inoculated mice was 4.1 compared to the mean infectivity score of 0.1 for trimethoprim/sulfamethoxazole (50/250 mg/kg) treated inoculated mice, approximately a four-log difference. An inoculated mouse model of P. carinii infection provides both a source of organisms from a different host and an animal model for study of drugs for therapy and prophylaxis which is less costly than rats and which requires less drug than required for rats.

Animals↗
PubMedDate not supplied

An ELISA method for quantitation of Pneumocystis carinii in culture and lung.

Numbers of Pneumocystis carinii in cultures or tissues traditionally have been determined by counting organisms on Giemsa-stained slides. For cultures, 10 microliters of culture supernatants have been sampled and counted on days 1, 3, 5 and 7. Infectivity scores of P. carinii-infected animal lung have been determined by three examiners scoring lung impression smears stained with Giemsa using a roughly logarithmic scale. Both counting procedures are tedious and time consuming. We have developed an enzyme-linked immunosorbent assay (ELISA) system which uses culture supernatants (in vitro) or homogenized animal lung (in vivo) as antigen, convalescent rat sera as primary antibody, and goat anti-rat alkaline phosphatase-conjugated immunoglobulin G as secondary antibody. The ELISA method shows good correlation with manual counts of Giemsa stains and allows a more rapid, more efficient method for quantitating P. carinii in both culture and infected lung.

Animals↗
PubMedDate not supplied

Inhibition of Pneumocystis dihydrofolate reductase by analogs of pyrimethamine, methotrexate and trimetrexate.

Dihydrofolate reductase was obtained from Pneumocystis carinii isolated from heavily infected lungs of female Sprague-Dawley rats infected by transtracheal inoculation. The enzyme differed significantly from other forms of dihydrofolate reductase in response to KCl and to antifolate drugs. Dihydrofolate reductase from P. carinii was used to assess activity of analogs of pyrimethamine, methotrexate, and trimetrexate. One pyrimethamine analog was selective for P. carinii dihydrofolate reductase; potency was in the micromolar range. In contrast, 21 methotrexate analogs and 2 trimetrexate analogs were selective for P. carinii dihydrofolate reductase; potencies for these were in the nanomolar range.

Animals↗