Biomedical subjects

J S Hunt

Publications and source records attributed to J S Hunt.

At least 145 records · Page 8Linked to original sources

PubMed1980

Isolation of human glomerular basement membrane antigens by affinity chromatography utilising Goodpasture's kidney antibody eluates.

The soluble material produced from a 96-hour digest of sonicated human glomerular basement membranes with purified collagenase was shown to contain at least 12 components by sodium dodecyl sulphate-polyacrylamide gel electrophoresis with molecular weights ranging from 20 x 10(3) to greater than 200 x 10(3) daltons. Five of these components produced antigenic peaks after two-dimensional immunoelectrophoresis into agar plates containing rabbit antiserum to the solubilised glomerular basement membrane. Two groups of antigenic components were demonstrated in these gels by two-dimensional immunoelectrophoresis autoradiographic techniques utilising 125I-labelled collagenase-digested human glomerular basement membranes reacted with antibodies eluted at acid pH from the kidney of the Goodpasture's patient. This acid eluate was shown to contain contaminants of alpha 1-antitrypsin and albumin co-eluting with the antibodies bound to the glomerular basement membrane. After removal of these contaminants, the antibodies were linked to cyanogen bromide-activated Sepharose and used to affinity purify four antigenic fractions from the collagenase-digested glomerular basement membrane. These fractions were eluted by 0.2 M glycine pH 2.8 and with 0.5 M ammonium thiocyanate and had molecular weights of 22-25, 43-45, 65-70 and 205 x 10(3) daltons. The smaller molecular weight components were shown to be common to both included and excluded fractions obtained by Ultragel AcA 44 chromatography of the digested material in 10 mM phosphate pH 8. This suggests that the larger molecular weight component would be an aggregate of a smaller component. Preliminary carbohydrate and amino acid analyses indicated that the different elution procedures elicited antigens with different chemical characteristics.

Anti-Glomerular Basement Membrane Disease↗

PubMed1980

Macrophages in NZB/NZW mouse glomerular disease.

Glomerular cultures from mice have been established and the various cells associated with them observed microscopically by phase contrast time-lapse cinematography, after histological staining and ultrastructurally with the transmission and scanning electron microscope. The most common cell types were large 80-200 microns diameter irregular polyhedral cells appearing as flattened plate-like structures morphologically, possibly of epithelial origin. Smaller 70-100 microns diameter fusiform cells possibly derived from either endothelium or mesangium were also often seen. A third cell type, small 10-20 microns diameter circular, motile and actively phagocytic was especially noticeable in cultures of hybrid New Zealand Black/New Zealand White mice but was not present in appreciable numbers in control cultures from strains without renal disease. This third cell type appeared to have Fc receptors, stained positively with non-specific esterase and by indirect immunofluorescence with specific rabbit anti-mouse macrophage serum and had the morphological appearance of a macrophage by scanning and transmission electron microscopy. It is concluded that cells of macrophage origin are associated with the immune complex glomerulonephritis in these hybrid mice.

Age Factors↗

PubMed1980

Biochemical and mitogenic properties of Tamm-Horsfall urinary glycoprotein.

Thirty separate preparations of human Tamm-Horsfall urinary glycoprotein (THP) prepared from pooled urine of normal individuals were assessed for the capacity to stimulate in vitro blast transformation with normal human peripheral blood lymphocytes. Detailed studies were performed on some of these preparations by various biochemical, ultrastructural and in vitro techniques. Heterogeneity amongst the sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE) profiles of several electrophoresis (2D-IEP). Gas liquid chromatographic analysis (GLC) revealed significant amounts of glucose in two of these samples. Ultrastructural studies using direct staining or platinum carbon shadowed specimens confirmed the fibrillar polymeric nature of soluble THP in vitro studies with human peripheral blood lymphocyte populations showed THP to have mitogenic properties towards B cells associated with the production of intracytoplasmic immunoglobulin. THP also stimulated the migration of mouse peritoneal exudate macrophages in a micro macrophages migration assay.

Cell Migration Inhibition↗

PubMed1979

A micro macrophage migration inhibition test for the detection of cellular immunity in vitro.

A simple, economical, macrophage migration inhibition assay enabling multiple tests to be performed using microculture plates has been developed. The system confers many advantages over the standard in vitro assay of macrophage migration inhibition factor. It requires only 5 X 10(5) macrophages for each assay, small volumes (100 microliter) of test factors are consumed, 96 tests in each culture plate are easily assessed microscopically and the technique is easily and reproducibly executed. An adherent monolayer of activated fresh mouse peritoneal exudate cells (PEC) is established in each microculture well. A 'starting line' is scored across the monolayer by the application of a trimmed razor blade and the cells are dislodged and aspirated from one side of the line. Test factors from cultures of stimulated lymphocytes were added to the 'wounded' monolayer and the migration of macrophages from test and control cultures was measured after staining. The cells across the line were counted after incubation. Miration was shown to be time and temperature dependent, markedly stimulated by colchicine and appeared to associate with tuberculin antigen (PPD) skin testing and lymphocyte transformation in human subjects.

Animals↗

PubMed1978

Tamm-Horsfall protein in the glomerular capsular space.

Tamm-Horsfall protein was detected within the capsular space by immunofluorescence in 7 of 72 consecutive patients on whom renal immunopathological studies were performed. Three patients showed prominent aggregates or crescentic collections affecting 30-50% glomeruli; the remaining four patients showed smaller aggregates between lobules. All patients showed pathological evidence of tubulointerstitial disease. It is suggested that Tamm-Horsfall protein in the capsular space is a sign of intratubular urinary backflow and that Tamm-Horsfall antiserum is a useful addition to the reagents used in the immunofluorescence study of renal biopsies.

Adult↗

PubMed1978

Stimulation of human peripheral blood lymphocytes by Tamm-Horsfall urinary glycoprotein.

Tamm-Horsfall urinary glycoprotein (THP), prepared by salt precipitation of pooled urine from normal individuals, stimulated purified human peripheral blood lymphocytes (PBL) to undergo blastoid transformation. the response was measured by tritiated thymidine uptake into DNA after 6 days in culture. Several batches of THP stimulated, in varying degrees, all samples of PBL tested and the response approached that seen with the mitogens phytohaemagglutinin (PHA), Concanavalin A (Con A) and pokeweed mitogen (PWM) after 4 days in culture. The response usually exceeded that seen after 6 days with tuberculin purified protein derivative (PPD) in Mantoux positive lymphocyte donors.

Concanavalin A↗

PubMed1977

On the cysteine and cystine content of proteins. Differences between intracellular and extracellular proteins.

Analysis of published data on the cysteine and half-cystine content of proteins indicates that most intracellular proteins may be classified as sulfhydryl proteins (those containing cysteine but little or no half-cystine) and that such sulfhydryl proteins have a low cysteine content. The mean systeine content found for 32 intracellular mammalian proteins was 1.6% and intracellular proteins of many bacteria have similar or lower values. Extracellular mammalian proteins are primarily disulfide proteins (those containing half-cystine but little or no cysteine) have a high half-cystine content, the mean value found for some 34 extracellular mammalian proteins being 4.1%. This is contrasted with many of the extracellular proteins from facultative bacteria which are cyst(e)ine-free proteins, being lacking in both cysteine and half-cystine. These and related observations are interpreted in terms of the evolution of life in a reducing atmosphere and the subsequent transition to an oxidizing environment. It is suggested that disulfide proteins evolved primarily after the accumulation of oxygen in the atmosphere.

Bacterial Proteins↗

PubMed1977

Kinetics and mechanisms of monolayer interactions IV: Surface activity of alkanols and energies of their interaction with dipalmitoyllecithin and dipalmitoylphosphatidylethanolamine.

The free energies of adsorption of the C1 to C14 alkanols at the air--water interface, estimated from plots of the surface pressure (pi less than or equal to 5 dynes/cm) against the bulk concentration, were a linear function of the chain length for 1-alkanols. From C3 to C8, the 2- and 3-isomers showed lower values than the 1-isomers. The energies of interaction of the C1 to C14 alkanols with dipalmitoylphosphatidylethanolamine and dipalmitoyllecithin monolayers, previously spread at the air-water interface, were estimated from the increase of the surface pressure with increasing concentrations of the injected alkanol. The energies of interaction of the C1 to C5 1-alkanols were linear functions of the chain length. The energy of interaction per methylene group of the alkyl chain suggests that the phospholipid monolayers behaved as ultrathin "oil" phases. The 2- and 3-isomers presented marked departures from linearity. The literature data for reflection coefficients in biomembranes and for partition coefficients between olive oil and water, red cell membranes and water, and phospholipid liposomes and water for the C1 to C5 alkanols show similar linearities with the chain length for the 1-isomers and comparable departures for the 2- and 3-isomers.

Adsorption↗

PubMed1977

Glomerular lesions in argyric NZB/NZW mice.

Glomerular basement membranes showed strong silver labelling on electron microscopy in NZB/NZW mice fed 12 mmol AgNO3 for 6 weeks at ages from 1-7 months. Although some silver granules were seen within mesangial immune deposits following their concurrent deposition, silver was usually deposited in electron-lucent areas around pre-existing deposits, or formed a boundary to an expanded mesangium when immune deposits subsequently developed. Immune deposits in capillary loops occurred on the outer or inner aspect of the zone of silver labelling.

Animals↗

PubMed1976

Silver deposition in mouse glomeruli.

Administration of 6 mM silver nitrate in the drinking water of mice resulted in deposition within the glomerular basement membrane of silver granules which were detected on electron microscopy after 12 days. Larger aggregates were detected in the basement membrane and mesangium when silver ingestion was extended to 14 weeks. The silver deposits did not alter significantly over a period of 21 weeks after silver nitrate ingestion was stopped.

Animals↗

PubMed1976

Immmune complex glomerular disease in argyric mice.

A mesangiopathic glomerulonephritis with prominent mesangial immunoglobulin deposits was induced in mice by single or multiple intraperitoneal injections of bovine serum albumin. The silver-labelled basement membrane in mice which had ingested 6mM silver nitrate in drinking water proved a useful marker indetermining that the immune deposits were on the intracapillary aspect of the basement membrane.

Animals↗

PubMed1974

BCG immunotherapy of malignant melanoma: summary of a seven-year experience.

Over the past 7 years, 151 patients with malignant melanoma have been treated with BCG immunotherapy alone or as an adjunct to surgical therapy. Direct injection of metastatic melanoma lesions limited to skin resulted in 90% regression of injected lesions and 17% regression of uninjected lesions in immunocompetent patients. Approximately 25% of these patients remained free of disease for 1 to 6 years. Direct injections of BCG into nodules of patients with subcutaneous or visceral metastases resulted in a lower incidence of local control and no long term survivors. Attempts to improve the results of immunotherapy in these patients by palliative surgical resection of large metastatic lesions to lower tumor burden followed by BCG immunotherapy significantly improved the results although many patients still developed recurrent disease. Early results of a clinical trial combining BCG immunotherapy with regional lymphadenectomy in patients with melanoma metastatic to lymph nodes have been encouraging and promising. Further controlled clinical trials are necessary to elucidate the role of BCG in immunotherapy. However, since BCG is but one of a number of potential immunologic adjuvants, even more effective immunotherapy will be possible as further knowledge of the interactions of cellular and humoral immunity is acquired.

Anaphylaxis↗

PubMed1974