Search PubMed⌕ Search

Biomedical subjects

E Reiss

Publications and source records attributed to E Reiss.

At least 55 records · Page 3Linked to original sources
PubMed1988

Serological, electrophoretic, and biological properties of Cryptococcus neoformans antigens.

We compared a cryptococcal culture filtrate antigen referred to as CneF with chemically defined cryptococcal antigen fractions isolated by Cherniak and co-workers by using double immunodiffusion gels, polyacrylamide gel electrophoresis, immunoblots, and footpad reactivity of immunized mice. The three previously described components of cryptococcal culture filtrates are a high-molecular-weight glucuronoxylo-mannan (GXM), which is the major constituent, a galactoxylomannan (GaIXM), and a mannoprotein (MP). In this study we demonstrated that CneF contained components which were serologically and electrophoretically similar to the three previously described cryptococcal culture filtrate fractions. The MP fraction elicited significantly stronger delayed-type hypersensitivity responses than did the GXM or GaIXM fraction when used in mice immunized either with the CneF in complete Freund adjuvant or whole heat-killed Cryptococcus neoformans yeast cells. These findings were confirmed when the footpads of immunized mice were challenged with GaIXM and MP preparations from a culture filtrate of a C. neoformans acapsular mutant that does not produce GXM. Thus, we concluded that the MP was the primary component recognized by the anticryptococcal cell-mediated immune response in mice.

Antibodies, Fungal↗
PubMed1988

Enhanced binding of capsular polysaccharides of Cryptococcus neoformans to polystyrene microtitration plates for enzyme-linked immunosorbent assay.

A sensitive enzyme-linked immunosorbent assay (ELISA) to measure antibodies against capsular polysaccharide was developed, based on the enhanced binding of polysaccharide to polystyrene microtitration plates. The wells of the microtitration plate were primed with an adipic acid dihydrazide derivative of bovine serum albumin (AH-BSA) (100 micrograms/mL, 0.01 M NaPO4-0.14 M NaCl, pH 7.2 (PBS]. Capsular polysaccharide, the glucuronoxylomannan of Cryptococcus neoformans serotype A, was oxidized with NaIO4 for 5 min; the reaction was then quenched with ethylene glycol. The partially oxidized polysaccharide was dialyzed vs. PBS, and its concentration was adjusted to 50 micrograms/mL with PBS. This solution (100 microL/well) was covalently bound to the AH-BSA primed microtitration plates through formation of a Schiff base between the hydrazide group on the AH-BSA and the aldehyde groups on the polysaccharide. Antimouse IgG-alkaline phosphatase conjugate was used in an indirect ELISA to measure captured murine monoclonal antibodies directed against glucuronoxylomannan. Mean absorbances, after 15 min, were 0.13 in negative control wells, and greater than 0.7 in test wells. No intermediate steps were required to block nonspecific binding of antibody.

Antibodies↗
PubMed1988

Establishment of primary cell cultures from human and canine parathyroid gland explants.

Primary parathyroid cell cultures were established from 22 canine and 20 human parathyroid glands explanted into Connaught Medical Research Laboratory-1415 (CMRL-1415) medium containing fetal calf serum (FCS). Initiation of cellular outgrowth and the rate of cellular propagation through the first subcultures were evaluated in various media. These included Roswell Park Memorial Institute-1640 (RPMI-1640), minimal essential medium (MEM), medium-199 (M-199) and Coon's modified Ham's F-12 (F12) and CMRL-1415 media. Only CMRL-1415 medium supplemented with FCS invariably supported the vigorous rate of cellular growth as determined by the number of cells in cultures, their viability, parathyroid hormone (PTH) levels in the growth media and the number of proliferating cells. Propagation of parathyroid cells in vitro was limited to five passages. Cessation of cell division was always coincidental with the emergence of numerous, non-dividing binucleated cells. These experiments demonstrate that primary parathyroid cell cultures from canine and human parathyroid gland explants can be readily established in CMRL-1415 medium containing FCS.

Animals↗
PubMed1988

DNA patterns in parathyroid disease predict postoperative parathyroid hormone secretion.

Flow cytometric analysis of the nuclear deoxyribonucleic acid (DNA) content of parathyroid glands excised from patients with hypercalcemic hyperparathyroidism has identified three distinct DNA patterns. The most frequent pattern showed a high percentage of cells with tetraploid DNA, which indicated an increase in the G2 and M phase of the cell cycle. Thirty-four patients were found to have abnormal tetraploid DNA content. One patient had a normal diploid pattern, and seven were found to have an aneuploid DNA population in their excised parathyroid glands. This unexpected finding of aneuploid DNA appears to be an unique feature of these endocrine glands because they have no histologic or clinical characteristics of malignant change. All patients have remained normocalcemic and clinically well after excision of only grossly enlarged glands. Postoperative parathyroid hormone (PTH) levels were correlated in 17 patients with DNA analyses of biopsy specimens from 30 normal-sized glands which were left in situ. Seven patients with elevated PTH postoperatively had high tetraploid or aneuploid DNA in all 13 glands from which biopsy specimens had high tetraploid or aneuploid DNA in all 13 glands from which biopsy specimens had been taken. In 10 patients with normal PTH levels, six had normal diploid patterns, whereas four had high tetraploid DNA in their gland biopsy specimens. DNA content present in biopsy specimens of normal-sized, in situ glands was predictive (p less than 0.042) of parathyroid gland secretory activity. These findings suggest that the stimulus for parathyroid gland hyperfunction often affects more than a single enlarged gland and persists after clinical cure, as shown by a more rapid cell turnover in some remaining glands and continued hypersecretion of hormone.

DNA↗
PubMed1987

An electrophoretic karyotype for Candida albicans reveals large chromosomes in multiples.

Using field-inversion gel electrophoresis we defined an electrophoretic karyotype for the yeast, Candida albicans. The karyotype is distinct from other species of Candida and is species specific. A total of five distinct chromosomal mobility groups were observed, at least four of which are composed of a minimum of two fragments each. From the apparent sizes of these fragments relative to the large chromosomes of the morphologically related yeast Saccharomyces cerevisiae, together with the known genome size of this organism, we conclude that the karyotype is the result of the migration of intact chromosomes.

Candida↗
PubMed1986

Ultrastructure of acapsular mutant Cryptococcus neoformans cap 67 and monosaccharide composition of cell extracts.

Acapsular mutant Cryptococcus neoformans cap 67 was grown in Pine's citrate broth medium for 3 days and the cells then transferred to a nitrogen-free medium for 6 days. The cells were subjected to a four stage extraction with buffered Triton-X100, cold dilute alkaline borohydride, hot dilute acetic acid, and a second alkaline extraction. Galactoxylomannan antigens were recovered from the culture supernates of both 3 day-old and 9 day-old yeast cells. The alkaline extracts contained water-soluble galactoxylomannan and a water-insoluble glucan. Dilute acid treatment released a minor amount of carbohydrate from the cells. The second alkaline extraction yielded increased amounts of glucan and galactoxylomannan from the 9 day-old cells. Soluble non-dialyzable cell extracts were antigenically identical in immunodiffusion with the culture supernate antigens. After the extraction sequence, all of the galactose, xylose, and mannose were removed from the cells. The walls retained their shape after extraction but their layers were loosened. Cells resuspended in nitrogen-free medium for six days developed thickened walls with alternating electron-dense and electron-lucent layers. The major constituent of the thickened 9 day-old cell walls was glucose, only 5% glucosamine was detected.

Antigens, Fungal↗
PubMed1986

Monoclonal antibodies against the M-protein and carbohydrate antigens of histoplasmin characterized by the enzyme-linked immunoelectrotransfer blot method.

Monoclonal antibodies (MAbs) of two different specificities were produced by immunizing mice with the semipurified M antigen of histoplasmin. One type, from clone CB4, was an immunoglobulin M that precipitated a polysaccharide present in histoplasmin and also formed immunoprecipitates with a cross-reactive polysaccharide present in extracts of Blastomyces dermatitidis and Coccidioides immitis. The second type of MAb, from clone EC2, was an immunoglobulin G that reacted in the enzyme-linked immunoelectrotransfer blot (EITB) assay with a doublet of proteins with an apparent molecular size of 70 to 75 kilodaltons. This molecule is proposed as the authentic M protein antigen that is recognized by M antibodies in sera from mice and rabbits immunized with Histoplasma capsulatum and from persons with histoplasmosis. The M factor also occurs in an abundant disulfide-bridged dimer which has a molecular size of 150 kilodaltons and is nonimmunoreactive under the conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Animals↗
PubMed1986

Monoclonal antibodies against Candida tropicalis mannan: antigen detection by enzyme immunoassay and immunofluorescence.

Three strains of mice were immunized with Candida tropicalis cell walls, and antibodies against mannan were detected by indirect enzyme immunoassay (EIA) in 3 of 9 BALB/c mice, 4 of 11 C57BL/6 mice, and 4 of 8 CFW mice. Responding mice produced immunoglobulin M (IgM), but IgG was not detected in their sera. Fusion of the high-responder BALB/c mouse with a plasmacytoma cell line resulted in 41 clones secreting antimannan monoclonal antibodies (MAbs). Four clones selected for propagation included one IgM and one IgG MAb that reacted with mannans of Candida albicans serotypes A and B and of C. tropicalis and two IgM MAbs specific for an epitope only in the mannans of C. albicans serotype A and C. tropicalis. One of the IgM MAbs, CB6, was an effective substitute for rabbit antibodies in the double-antibody sandwich EIA to detect antigenemia produced in rabbits infected with C. albicans A or C. tropicalis. It could function either as the peroxidase-conjugated indicator antibody or as the capture antibody. Two MAbs, CB6 (C. tropicalis and C. albicans A specific) and AC3 (C. tropicalis and C. albicans A and B specific), functioned in place of polyclonal antisera in the serotyping of C. albicans by immunofluorescence. There was 95.8% agreement in the results of serotyping using MAbs as reagents compared with rabbit antisera. Competitive inhibition in EIA between CB6 and monospecific antisera against C. albicans factors 1, 4, and 6 indicated that CB6 binds to an epitope which is probably factor 6. Serologic similarity between factor 4 and the binding site of MAb AC3 was also determined.

Animals↗
PubMed1986

Monoclonal antibodies against isoelectrically focused Nocardia asteroides proteins characterized by the enzyme-linked immunoelectro-transfer blot method.

Sera from rabbits immunized with culture filtrates and homogenates of Nocardia asteroides B1042 gave at least eight precipitin bands by immunoelectrophoresis. At least 20 proteins with isoelectric points (pls) in the pH 4 to 5.4 range were observed in isoelectric focusing patterns. The enzyme-linked immunoelectro-transfer blot (EITB) assay showed that several of the isofocused proteins reacted with rabbit antisera and with sera from nocardiosis and tuberculosis patients. Antibodies against three proteins with pls of 4, 4.43, and 4.68 (antigenic factors 1,6,8) were present in nocardiosis patients' sera. The proteins were excised from isofocused gels, and IgG monoclonal antibodies (MAbs) were produced by the hybridoma method. MAbs against factors 1 and 6 did not crossreact with cytoplasmic antigens of Mycobacterium chelonae, M. intracellulare serotypes 4B and 8A, M. fortuitum, M. gordonae, or M. kansasii in the EITB method. Factor 8 (MAb) crossreacted with antigens of M. intracellulare and M. fortuitum.

Animals↗
PubMed1986

Antigenemia in rabbits infected with Candida albicans serotype B: detection by enzyme immunoassay and preliminary characterization of the antigen.

Peroxidase-labeled IgG against Candida albicans serotype A cell walls failed to detect antigenemia in rabbits infected with C. albicans type B. Therefore type B antisera were produced in rabbits immunized with either type B cell walls; whole blastoconidia heated and killed at 60 degrees C (HK); a sublethal intravenous dosage of 10(6) colony forming units (c.f.u.); or a normally lethal dose of 10(7) c.f.u. injected into subcutaneously implanted plastic chambers. The four antisera were conjugated to peroxidase for a double antibody sandwich enzyme immunoassay. Sixteen rabbits, immunosuppressed with cortisone, received 10(7) type B blastoconidia intravenously. On day 4 their kidneys contained between 3.4 X 10(8) and 1.4 X 10(11) c.f.u. Antigenemia was detected only with the conjugated anti-HK-IgG. A mean concentration of 246 ng ml-1 of circulating antigen was detected only when serum-antigen complexes were dissociated by boiling in EDTA. The properties of the circulating antigen were as follows: relative molecular weight between 50 000 and 100 000 da; stable to boiling and proteinases, labile to sodium periodate oxidation, and bound by concanavalin A. These properties are consistent with a polysaccharide or glycoprotein, probably the mannoprotein of the cell wall. Antigenic specificity was probed further using IgG produced against heat-killed C. albicans serotype A. Antigenemia resulting from infection with serotype B was detected in 5 of 5 rabbits tested with the anti-HK-serotype B IgG but only with 3 of 5 tested with the anti-HK-serotype A IgG. In addition, the concentrations detected with serotype B reagents were 31 times higher.

Animals↗
PubMed1986

The parathyroid gland in health and disease.

The authors studied the parathyroid glands from 100 previously healthy subjects who died suddenly and were admitted to the Dade County Medical Examiner's (ME) morgue and from 66 inpatients who died at Jackson Memorial Hospital (JMH). Parathyroid glands in patients with diseases (JMH series) were heavier than those in healthy persons (ME series), and both groups of glands were significantly heavier than those previously reported. Mean glandular weight in white subjects was 42.6 +/- 20.3 mg, with a range of 22-103 mg. The 95% upper limit of gland weight for healthy white subjects was 73.1 mg and for black subjects, 91.6 mg. The size and weight exhibited a skewed distribution. Gland weight varied with age, increasing to a maximum in the 41-60 year old age group in all subsets except white women, in whom it continued to increase till after age 70. There was slight correlation (r2 = 0.15) of gland weight with body weight within series and race groups; parenchymal content of the glands was not constant but correlated positively with glandular weight. Glands from both series had a comparable fat content. Fat was unevenly distributed throughout the gland, and its amount was highly variable, ranging between 0 and 90%, with a mean of 26% for white subjects and 24% for black subjects in both series. Therefore, percentage fat may not be used as an index of hyperplasia. Healthy back subjects had heavier glands than healthy white subjects, unaccounted for by differences in body weights; this difference was not statistically significant in subjects with disease. Within the black race, glands were not significantly heavier in disease than in health, and in the few cases with serum calcium determinations, the gland weight did not vary inversely with serum calcium levels as in white subjects, suggesting a basic difference in parathyroid calcium metabolism between the two races.

Adolescent↗
PubMed1985

Characterization of protein and mannan polysaccharide antigens of yeasts, moulds, and actinomycetes.

Antigens in coccidioidin were compared with purified subfractions via tandem immunoelectrophoresis (IEP) and by a combination of advancing line and crossed IEP. Rocket IEP was suitable for titrating the reactions and showing the relationship between column fractions. These techniques required multicomponent antisera produced by hyperimmunization over many months and by the use of known standard migration pairs. The IEP variations were used to chart the development of antisera against coccidioidin factors, to monitor antigen purifications, and to test the immunochemical homogeneity of an isolated antigen. Mannose-based heteroglycans of Cryptococcus neoformans were recovered from the culture filtrate. After precipitation of the major viscous glucuronoxylomannan (GXM) with ethanol or cetyltrimethylammonium bromide, the supernate is reserved because it contains a galactoxylomannan (GalXM). After removal of glucuronic acid from the GXM, the resulting xylomannan of serotype A was amenable to 13C-nuclear magnetic resonance (NMR) spectrometry; it revealed nonreducing xylose, alpha-1,3-mannose, and alpha-1,2/1,3 disubstituted mannose, thus confirming by an independent means what was previously known. The characterization sequence of GalXM included: (1) gas-liquid chromatography (GLC) of neutral sugars as peracetylated aldononitriles; (2) methylation-fragmentation GLC mass spectrometry to determine the glycosidic linkages; and (3) 13C-NMR showing similarities to mannan of Saccharomyces cerevisiae. Affinity chromatography of the GalXM on concanavalin A separated the galactoxylo component from an adsorbed mannoprotein. Selection of monoclonal antibodies (MAbs) relies on presumptive enzyme immunoassays (EIAs) or radioimmunoassays for rapid screening of clones and for determination of isotypes; however, higher resolution confirmatory tests are needed to obtain MAbs of desired specificity. MAbs against Candida tropicalis mannan were labeled with horseradish peroxidase to use for detecting mannan in serum. MAbs against the partially purified "m" factor of histoplasmin were characterized by the enzyme-linked immunoelectro-transfer blot technique (EITB), revealing unsuspected complexity in the antigen. Secreted proteins of Nocardia asteroides were isoelectrically focused; three proteins, identified by EITB as promising to be specific for that actinomycete, were cut out of gels and used to immunize mice for production of MAbs. The fimbriae of Actinomyces viscosus and A. naeslundii that mediate lactose-reversible coagglutination with Streptococcus sanguis have been used to evoke MAbs.(ABSTRACT TRUNCATED AT 400 WORDS)

Actinomycetales↗
PubMed1985

Comparison of enzyme immunoassay and gas-liquid chromatography for the rapid diagnosis of invasive candidiasis in cancer patients.

Three proposed quantitative markers for candidiasis, arabinitol, mannose, and mannan in serum, are compared in 50 normal blood donors and 38 high-risk patients, 23 with and 15 without invasive candidiasis. Arabinitol concentrations in serum, the arabinitol/creatinine ratio, and mannose concentrations in serum were significantly greater in the 15 patients without candidiasis than in the normal blood donors (P less than 0.05). The sensitivities and specificities were 26 and 87% for arabinitol, 13 and 93% for the arabinitol/creatinine ratio, and 39 and 87% for mannose. On the other hand, mannan concentrations in serum were less than 1 ng/ml in normal blood donors and patients without candidiasis (P = 0.344), and the sensitivity and specificity were 65 and 100%, respectively. Of 23 patients with proven or probable candidiasis, 16 had mannan levels in serum greater than the mean + 2 standard deviations (0.46 ng/ml) for the 15 controls. In 16 patients with invasive candidiasis and positive blood cultures for the Candida spp., only 13 had elevated levels of at least one of the three markers. The arabinitol/creatinine ratio, the mannose level, and the mannan level became elevated an average of 4 days before, 1 day before, and on the same day that the blood cultures were drawn, respectively. Conversely, mannan was detected in the sera of six of seven patients with invasive candidiasis and negative blood cultures. We conclude that the best approach to diagnosing invasive candidiasis involves obtaining blood cultures and carrying out serial assays for mannan in serum.

Candidiasis↗
PubMed1984

Oncogenous osteomalacia. Review of the world literature of 42 cases and report of two new cases.

The clinical and biochemical data in 42 reported cases of oncogenous osteomalacia are reviewed, and data in two previously unreported cases are recorded. It is likely that the syndrome is more common than suggested by the paucity of reports and may account for a substantial fraction of nonfamilial, adult-onset "idiopathic" osteomalacia. Tumors associated with the syndrome are characteristically benign, of mesenchymal origin, highly vascular, and composed principally of giant and spindle cells. Complete excision of tumors results in cure of the osteomalacia in the majority of patients. The syndrome presumably represents an instance of humor-induced phosphaturia but supporting experimental data are scanty. Plasma levels of 1,25-dihydroxycholecalciferol are uniformly low, and treatment with this metabolite is generally very effective; however, abnormal vitamin D metabolism cannot by itself account for the syndrome.

Adolescent↗
PubMed1984

Comparison of serum mannan, arabinitol, and mannose in experimental disseminated candidiasis.

Concentrations of arabinitol, mannose, and mannan in serum have independently been reported to be elevated in patients with invasive candidiasis. These three marker substances were compared in a rabbit model. Twelve rabbits, immunosuppressed with cortisone, were infected intravenously with Candida albicans 3181A. Six uninfected control animals also received cortisone, and four rabbits were neither infected nor immunosuppressed. Blood samples, drawn from 2 days before to 14 days after infection, were assayed for serum mannan by sandwich enzyme immunoassay, antibodies to mannan by indirect enzyme immunoassay, arabinitol and mannose by gas-liquid chromatography, and serum creatinine. Serum mannan, negative before infection, peaked in all infected animals 4 days after infection (mean, 18 ng/ml) and decreased thereafter. Significant increases (2 standard deviations greater than mean in normals) in arabinitol, the arabinitol/creatinine ratio, and mannose were found in 12, 8, and 12 of the infected rabbits, respectively, but also in all 6 uninfected animals receiving cortisone. Only serum mannan was specific in this immunosuppressed rabbit model.

Animals↗