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Biomedical subjects

J C Rhodes

Publications and source records attributed to J C Rhodes.

At least 37 records · Page 2Linked to original sources

Elevated Accumulation of Proline in NaCl-Adapted Tobacco Cells Is Not Due to Altered Delta-Pyrroline-5-Carboxylate Reductase.

Tobacco (Nicotiana tabacum L. var Wisconsin 38) cells that are adapted to 428 millimolar NaCl accumulate proline mainly due to increased synthesis from glutamate. These cells were used to evaluate the possible role of Delta(1)-pyrroline-5-carboxylate reductase in the regulation of proline biosynthesis. No increase in the specific activity of Delta(1)-pyrroline-5-carboxylate reductase in crude extracts throughout the growth cycle was observed in NaCl-adapted cells compared to unadapted cells. The enzyme from both cell types was purified extensively. On the basis of affinity for the substrates NADPH, NADH, and Delta(1)-pyrroline-5-carboxylate, pH profiles, chromatographic behavior during purification, and electrophoretic mobility of the native enzyme, the activities of the enzyme from the two sources were similar. These data suggest that the NaCl-dependent regulation of proline synthesis in tobacco cells does not involve induction of pyrroline-5-carboxylate isozymes or changes in its kinetic properties.

Journal Article↗

Antigenicity and immunogenicity of an extract from the cell wall and cell membrane of Histoplasma capsulatum yeast cells.

In order to identify T-cell antigens from Histoplasma capsulatum yeast cells, we prepared a detergent extract of the cell wall and cell membrane of yeast-phase H. capsulatum G217B and analyzed its antigenicity and immunogenicity. Mice injected with viable H. capsulatum yeast cells or with 500 or 1,000 micrograms of the extract mounted a delayed-type hypersensitivity response to solubilized cell wall and cell membrane. Vaccination with this antigenic preparation conferred a protective immune response in mice that were challenged intravenously with H. capsulatum yeast cells. The extract induced in vitro proliferation by splenocytes from mice injected with either viable yeast cells or the soluble cell wall and cell membrane preparation. We also examined the profile of in vitro responses by a murine T-cell line and by cloned T cells to soluble cell wall and cell membrane by employing the technique of T-cell immunoblotting. Two prominent regions that stimulated the T-cell line and cloned T cells were identified. Fractions encompassing an area between 53 and 64 kDa caused proliferation by a T-cell line and five of six clones. Antigens recognized by the T-cell line and by three of six clones were contained in another area that extended from 69 to 82 kDa. The data demonstrate that this soluble extract from cell wall and cell membrane contains antigens recognized by T cells and mediates protective immunity. Moreover, T-cell immunoblotting provides a useful technique for mapping immunoreactive molecules from H. capsulatum yeast cells.

Animals↗

The elastinolytic proteinase of Aspergillus flavus is not glycosylated.

Aspergillus flavus produces an elastinolytic metalloproteinase in culture fluid which does not appear to be a glycoprotein. The elastase did not stain with periodic acid Schiff reagent, its migration rate was not changed by digestion with glycosidases or chemical agents, and its release into culture medium was not inhibited by tunicamycin.

Animals↗

Experimental murine invasive pulmonary aspergillosis.

A new model of invasive pulmonary aspergillosis (IPA) was developed in immunosuppressed mice. Intranasal route of inoculation was used to deliver predictable numbers of Aspergillus flavus conidia. The LD50 was determined to be 2.7 X 10(2) viable conidia, and a combination of quantitative culture and determination of chitin content was shown to best measure the progression of pulmonary disease. The evolution of IPA in these mice conformed with what has been reported in human cases of aspergillosis; both histopathology of the pulmonary lesions and dissemination pattern resembled their human counterparts. The authors hope to use this model to study virulence mechanisms of Aspergillus and novel therapeutic methods.

Administration, Intranasal↗

Isolation and characterization of an elastinolytic proteinase from Aspergillus flavus.

An elastinolytic proteinase of Aspergillus flavus has been isolated to homogeneity, and its physical and biochemical properties have been characterized. Two purification protocols were compared; an initial step of ion-exchange chromatography was found to be equivalent to ammonium sulfate precipitation at neutral pH. A combination of gel filtration and adsorption chromatographies on the resultant crude enzyme produced highly purified elastase with yields of 5 to 10%. The enzyme is a 23-kilodalton protein with a pI of 7.6. The enzyme activity is markedly inhibited by numerous metal ions. Aspergillus elastase appears to be a metalloproteinase EC 3.4.24.X), as determined by its sensitivity to 1,10-phenanthroline.

Aspergillus flavus↗

Development of two isogenic sweet corn hybrids differing for glycinebetaine content.

A hybrid of sweet corn, Zea mays L. (;1720'; Rogers Brothers Seed Co.), was found to be comprised of glycinebetaine-positive and glycinebetaine-deficient individuals in a 1:1 mixture. This phenomenon was traced to segregation for a single, nuclear, dominant gene determining leaf glycinebetaine content within the female inbred parent of this hybrid. Selection for homozygous recessive (glycinebetaine-deficient) and homozygous dominant (glycinebetaine-positive) genotypes of the female inbred parent enabled production of two isogenic versions of hybrid ;1720' differing with respect to a single copy of the dominant allele, by mating these female parent selections with the common homozygous recessive (glycinebetaine-deficient) male parent. These two isogenic hybrids are shown to differ by a factor of 300- to 400-fold in glycinebetaine titer of young expanding leaves of salinized plants, but exhibit no striking differences in the levels of free amino acids or the level of N-methylnicotinic acid (nicotinic acid betaine; trigonelline). The only significant difference between the two hybrids in terms of amino acid composition was found to be in the level of alanine under nonsalinized conditions. The betaine-deficient hybrid exhibited a 14% lower alanine level than the betaine-positive hybrid. Betaine deficiency was not associated with altered stress-induced accumulation of amino acids such as proline, serine, and asparagine plus aspartate, attesting to the high specificity of the genetic difference between these isogenic hybrids with respect to betaine accumulation. This germplasm offers unique opportunities to test whether a single dominant allele determining stress-induced betaine accumulation capacity influences stress resistance in maize.

Journal Article↗

Paecilomyces variotii. An unusual cause of isolated sphenoid sinusitis.

Isolated fungal sphenoid sinusitis is an unusual head and neck infection. We describe the first reported case (to our knowledge) of Paecilomyces variotii sphenoid sinusitis. In addition, it represents one of the few reported cases of isolated fungal sphenoid sinusitis. Diagnosis and management of this condition are described. Aggressive surgical management is advocated when fungal sphenoid sinusitis is suspected.

Aged↗

Elastase production in clinical isolates of Aspergillus.

Clinical isolates of Aspergillus species were tested, both retrospectively and prospectively, for elastase activity using rose bengal-elastin agar plates. Patient records were then reviewed to determine the clinical diagnosis including the presence or absence of aspergillosis. All isolates that caused invasive aspergillosis produced elastase, but not all isolates producing elastase were associated with invasive disease.

Aspergillosis↗

Virulence factors in fungal pathogens.

Fungal pathogens possess virulence factors that allow them to cause disease in humans and animals. Combinations of properties such as temperature tolerance, evasion of host defences, dimorphism, and enzymatic activities may serve as virulence mechanisms for fungi.

Animals↗

Encapsulation and melanin formation as indicators of virulence in Cryptococcus neoformans.

Acapsular (Cap-) mutants of Cryptococcus neoformans var. neoformans that produce melanin (Mel+) on diphenol media at 30 degrees C but not at 37 degrees C were found to be avirulent for mice. Compared with wild-type isolates, the mutants had a lower rate of L-3,4-dihydroxyphenylalanine uptake at 37 degrees C and showed an insignificant level of phenoloxidase activity at both temperatures. To study the relationship of Cap and Mel phenotypes to virulence in mice, we crossed one of the mutants (Cap- Mel-) with a wild type (Cap+ Mel+) to obtain four classes of progeny (Cap+ Mel+, Cap+ Mel-, Cap- Mel+, and Cap- Mel-). The progeny with the Cap+ Mel+ phenotype and the wild-type parent (Cap+ Mel+) were inoculated into mice (10(6) cells per mouse) and, within 40 days, produced fatal infection in 90 to 100% of the animals. None of the other three phenotypes produced fatal infection within the same period. While progeny with the Cap+ Mel- phenotype did produce fatal infection after 40 days, 70 to 90% of the mice survived at least until day 70. However, in the isolates recovered from the brain tissue of a mouse that died on day 68, nearly 40% of the CFU had reverted to the Cap+ Mel+ type. The virulence of one of these revertant Cap+ Mel+ isolates was compared with that of a Cap+ Mel- isolate recovered from the same tissue. One hundred percent of the mice inoculated with the revertant died within 35 days, while no fatal infection was produced in the mice inoculated with the Cap+ Mel- isolate within the same period. The isolates with the Cap- Mel+ or Cap- Mel- phenotype not only failed to produce fatal infection but failed to revert to the Cap+ Mel+ type in the mouse brain during the experimental period. These results indicate that both the Cap+ phenotype and the Mel+ phenotype are important indicators of virulence in C. neoformans.

Animals↗

Contribution of complement component C5 to the pathogenesis of experimental murine cryptococcosis.

C5-deficient (C5-) mice succumb much sooner after intravenous inoculation with Cryptococcus neoformans than do C5-sufficient (C5+) mice. The C5- mice developed acute, fatal cryptococcal pneumonia, whereas the C5+ mice did not. The pneumonia was characterized by lung viable counts in C5- mice up to 1000-fold higher than in C5+, initial sequestration of twice as much 59Fe-labeled C. neoformans, and subsequent development of pulmonary edema. Chemotaxis of heterophils (PMNs) and mononuclear cells in response to C. neoformans was markedly greater in C5+ mice than in C5- animals. The effect of C5 on localization and growth of C. neoformans in the lung appeared to account for the disparate survival times of C5+ and C5+ mice after intravenous inoculation with C. neoformans.

Animals↗

Production and regeneration of protoplasts from Cryptococcus.

Protoplasts were quickly and efficiently produced from both varieties of Cryptococcus neoformans and from C. laurentii by use of the multi-enzyme product Novozym 234. Conditions for regeneration of protoplasts are described. DNA yield from the Novozym-produced protoplasts was superior to that from snail gut enzyme-derived protoplasts.

Cryptococcus↗

Inhibition of antipyrine metabolite formation in rats in vivo.

The effect of four inhibitors of cytochrome P-450-mediated drug oxidations (SKF 525A, cimetidine, metyrapone and alpha-naphthoflavone) on the urinary metabolite pattern and 14CO2 exhalation rate (CER)-time profile following [N-methyl-14C]antipyrine administration has been investigated. The CER-time profiles indicated that inhibition of antipyrine metabolism was in the rank order SKF 525A greater than cimetidine greater than metyrapone greater than ANF. The urinary metabolite patterns showed selectively in action towards particular pathways, 3-hydroxylation being primarily decreased by SKF 525A and cimetidine, and N-demethylation by ANF. The results provide further evidence for involvement of multiple forms of cytochrome P-450 in antipyrine metabolism.

Animals↗

Gonadal hormone regulation of MAO and other enzymes in hypothalamic areas.

Activities of type A monoamine oxidase (MAO), acetylcholine esterase (AChE), and glucose-6-phosphate dehydrogenase (G6PDH) were differentially altered in hormone-sensitive areas of the preoptic-hypothalamic continuum after administration of estrogen and progesterone. Estrogen increased activity of AChE in the bed nucleus of the stria terminalis and activity of G6PDH in the periventricular area (PVE) of the preoptic area, arcuate-median eminence (Ar-ME) and pituitary. Estrogen decreased activity of MAO in the PVE of the anterior hypothalamus, pars lateralis of the ventromedial nucleus and in the Ar-ME. Acute administration of progesterone (1 h) to estrogen-treated females did not further alter estrogen-dependent changes in AChE or G6PDH; however, MAO activity in the ventromedial nucleus and Ar-ME was rapidly increased after progesterone. Without prior estrogen administration, progesterone did not affect MAO activity. Administration of the protein synthesis inhibitor anisomycin prior to progesterone did not antagonize progesterone-dependent increases in MAO. Progesterone added in vitro to homogenates from estrogen-treated but not from untreated females increased MAO activity. The hormonal specificity, time course of effects and anatomical location of the enzymatic changes suggest that some of them may participate in the mediation of gonadal hormone action in the CNS. In particular, changes in MAO activity in the ventromedial nucleus and Ar-ME are consistent with reported effects of these hormones on monoamine turnover which in turn have been suggested to contribute to hormonal regulation of feminine sexual behavior and gonadotropin secretion.

Acetylcholinesterase↗

Taxonomic studies on Filobasidiella species and their anamorphs.

The taxonomy of Filobasidiella neoformans Kwon-Chung and F. bacillispora Kwon-Chung and their anamorphs were reinvestigated. Although the cross between the type culture of the two species failed to produce viable basidiospores, another pair of isolates did yield viable basidiospores. The segregation of phenotypic markers among the tetrads isolated from this inter specific cross proved that meiosis had occurred. On the basis of other previously known differences and the present genetic study, the two species are now considered to be two varieties of the species, F. neoformans. The anamorph of F. neoformans var. neoformans grew well at 37 degrees C in vitro and produced fatal infection in mice while that of F. neoformans var. bacillispora grew poorly at 37 degree C and failed to produce fatal infection in mice. Cryptococcus bacillisporus Kwon-Chung et Bennett is regarded as a synonym of C. neoformans var. gattii Vanbreuseghem et Takashio.

Crosses, Genetic↗

Interpretation of CO2 exhalation rate data from demethylation of aminopyrine and its metabolite monomethylaminoantipyrine.

1 Aminopyrine breath tests make use of the commercially available (N-dimethyl-[14C])-aminopyrine. A pharmacokinetic model has been proposed to relate 14CO2 exhalation rates (CER) to the demethylation of ([14C]-methyl)-aminopyrine (AP) and -monomethylaminoantipyrine (MAP). 2 computer simulations based on the model show that the shape of the CER-time profile is largely dependent on the ratio of the MAP to AP elimination rate constants. If this ratio equals 0.5 then the CERs decline in the monoexponential fashion. Ratios less than 0.5 result in concave biexponential curves whereas ratios greater than 0.5 result in convex curves. When demethylation is not complete for both compounds the transfer from biexponential to monoexponential behaviour will only occur at ratios greater than 0.5. 3 The resolution of concave biexponential CER-time profiles to give accurate estimates of AP and MAP elimination rate constants can only be achieved when the length of the experiment is adequate. The commonly employed 2 microCi tracer dose of aminopyrine is insufficient to monitor CER over the necessary time period to detect the proposed biexponential decline.

Aminopyridines↗

Phenoloxidase activity and virulence in isogenic strains of Cryptococcus neoformans.

A naturally occurring Mel- variant of Cryptococcus neoformans was isolated from the wild type. The effect of phenoloxidase activity on virulence was analyzed on genetically constructed Mel+ and Mel- isolates. The traits Mel+ and virulence in mice, as measured by cumulative mortality and replication potential in brain tissue, cosegregated among the progeny of a Mel+ X Mel- cross. Revertants (MelR) isolated during the course of the cumulative mortality experiment were used to compare virulence in isogenic sets of Mel- and MelR. In two separate sets of such isolates, Mel+ phenotype and virulence coreverted. Measurement of substrate uptake and phenoloxidase activity showed that loss of detectable phenoloxidase was the basis for the Mel- phenotype and that enzyme activity reappeared in the MelR isolates. An intermediate phenotype, Melbg, was also described. Cosegregation and coreversion of the melanin phenotype and virulence suggest that phenoloxidase is a virulence factor in C. neoformans.

Catechol Oxidase↗