Pneumocystis carinii pneumonia. An approach to rapid laboratory diagnosis.

Laboratory procedures used to establish the diagnosis of Pneumocystis carinii pneumonia were evaluated using an experimental murine model. Touch preparations and suspension smears were prepared from lung tissue know to contain Pneumocystis cysts. These preparations were stained by a variety of methods known to demonstrate either cyst forms or sporozoites and trophozoites. Suspension smears proved to be superior to touch preparations in terms of cyst content and homogeneity of staining. Also, methods that stain cyst forms were superior to those that stain sporozoites and trophozoites for location and identification of organisms. The authors believe that suspension smears prepared from lung tissue and stained with toluidine blue O should be examined initially as a rapid screening method for Pneumocystis cysts. When the results of this initial screen are negative or inconclusive, additional suspension smears stainded by the modified Gomori methenamine silver nitrate method should be examined, pending availability of histologic sections.

Antioxidants reduce the mutagenic effect of malonaldehyde and beta-propiolactone. Part IX. Antioxidants and cancer.

Increasing concentrations of malonaldehyde and beta-propiolactone were increasingly mutagenic with 7 mutants of Salmonella typhimurium, 5 of which mutated bya frameshift mechanism and 2 of which mutated through base-pair substitution. The antioxidants vitamin C, vitamin E, selenium and butylated hydroxytoluene (BHT) at 3 logarithmic concentrations markedly reduced mutagenesis in those strains which mutated by frameshift mechanism.

Cytokinin affects nuclear- and plastome-encoded energy-converting plastid enzymes.

Cytokinins induce two specific morphological alterations in mosses: (i) the differentiation of a tip-growing cell into a three-faced apical cell (the so-called bud), and (ii) the division of chloroplasts. In a developmental mutant of the moss Physcomitrella patens (Hedw.) B.S.G. (mutant PC22) impeded in both cellular differentiation (bud production) and chloroplast division, addition of cytokinin (N6-delta 2-isopentenyladenine) led to bud production after 3 d in the wild type and after 7 d in the mutant. Hormone induced a division of the mutant macrochloroplasts starting within 24 h and ongoing for 72 h. During this period the abundances of several plastid proteins changed in both genotypes as judged by two-dimensional-protein gel electrophoresis, silver staining and subsequent quantification with novel computer software. Eight of these polypeptides were isolated independently, subjected to microsequencing and thus identified, resulting in the first protein sequence data from a moss. Three polypeptides (24 kDa, 22 kDa, 20 kDa) were found to be homologous to enhancer protein OEE2 of the oxygen-evolving complex, four to represent isoforms of phosphoglycerate kinase (EC 2.7.2.3), and one was identified as the beta-chain of chloroplast ATPase (EC 3.6.1.34). Possible involvement of these key enzymes of the chloroplast energy-conversion machinery in organelle division and in cellular differentiation is discussed. Further sequence information was obtained from both subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39). Amounts of these polypeptides were not appreciably affected by cytokinin in moss chloroplasts.

Effects of blood on blood culture medium.

The morphological and biochemical changes that occur after inoculation of sterile blood into a blood culture medium (tryptic soy broth) with sodium polyanetholesulfonate and CO(2) were investigated. Cellular changes, pH, PCO(2), and PO(2) were monitored and evaluated. Erythrocytes became crenated and developed precipitated hemoglobin inclusions within 4 h. The lymphocytes appeared morphologically intact at 24 h, and by 48 h a few cells had undergone transformation. Many neutrophils were vacuolated at 24 h. Neutrophils capable of phagocytizing Staphylococcus aureus were observed after 18 h of incubation. Identifiable eosinophiles were present on day 6 of the study. A decrease in PO(2) in the unvented bottles from 44.4 to 8 mm of Hg occurred by 24 h. PO(2) remained low for 6 days, after which a slight increase occurred. An increase in PO(2) in the vented bottle from 51 to 58 mm of Hg occurred by 24 h of incubation. In both the vented and unvented bottles the PCO(2) increased. This increase was markedly more rapid in the unvented bottle. From a pH of 7.06 a decrease occurred for the first 24 h after inoculation, with the pH stabilizing at 6.8 in the vented bottles and at 6.6 in the unvented bottles. The biochemical changes that occurred in the vented culture bottles stabilized more rapidly than those of the unvented bottles. Changes caused by the addition of sterile blood to a blood culture medium resulted in conditions which departed considerably from accepted optima for the isolation of clinically important microorganisms. The phagocytosis of organisms that occurred may also have reduced the yield.

The plastome-encoded zfpA gene of a moss contains procaryotic as well as eucaryotic promoter consensus sequences and its RNA abundance is modulated by cytokinin.

Plastid DNA of the moss Physcomitrella patens has been sequenced. An open reading frame (ORF 315) was identified downstream from rbcL, between trnR-CCG and psaI. This ORF shares homology with zfpA, a putative regulatory gene in Pisum sativum. The moss ORF is preceded by a Shine-Dalgarno sequence, two plastid promoter consensus sequences, and three TATA boxes. A specific probe detected three transcripts of low abundance in the wild-type moss and a cytokinin-sensitive chloroplast mutant. Steady state levels of zfpA transcripts were different in the two genotypes. In mutant protonemata treated with cytokinin, steady state levels of the largest transcript decreased significantly.

False-negative stool occult blood tests caused by ingestion of ascorbic acid (vitamin C).

In a female patient with unexplained anemia, ascorbic acid ingestion and apparent false-negative occult blood tests were related. When she stopped ascorbic acid, her stools became strongly reactive ("4+") by three tests for occult stool blood; this association was observed repeatedly. A test developed to measure stool occult blood in the presence of ascorbic acid remained reactive throughout this observation, and the observation was confirmed by in-vitro studies. Current tests for occult blood depend on the pseudoperoxidase activity of heme and are inhibited by low levels of ascorbic acid. Reducing substances chemically similar to ascorbic acid also inhibits occult blood tests; oxidized ascorbic acid and sulfhydryl reducing agents do not inhibit them at physiologic levels.

Intrathyroidal parathyroid adenoma or hyperplasia. An occasionally overlooked cause of surgical failure in primary hyperparathyroidism.

In six surgically proved intrathyroidal parathyroid tumors, neither thyroid scan nor gross appearance of the thyroid at surgery allowed reliable detection of parathyroid glands wholly within the thyroid. Preoperative selective catheterization of thyroid veins with radioimmunoassay for parathyroid hormone facilitated successful removal of hyperfunctioning intrathyroidal parathyroid tissue in all five cases in which it was performed.