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Biomedical subjects

J R Archer

Publications and source records attributed to J R Archer.

At least 19 recordsLinked to original sources

Cell proliferation within the growth plate of the tich mouse.

The pattern of chondrocyte proliferation was studied in the proximal tibial growth plate of tich mice (gene symbol tch), a recessive mouse mutant, which is coisogenic with the A.TL strain. Specimens were qualitatively studied at time points of 2, 3, 4, 6, and 8 weeks of age by bromodeoxyuridine labeling of cell division and routine histology. At 2 weeks, when the lesion appeared as a full-width thickening of the growth plate, a greater proportion of cells appeared positively labeled in the proliferative zone. This concavity in the central portion of the growth plate became progressively more focal between 3 and 4 weeks to give a "tongue" of unresorbed, noncalcified cartilage in the central region of the tich growth plate. BrdUrd labeling indicated that the appearance of the cartilage tongue corresponded with increased cell division in the central region of the growth plate. At the same time, a "second" zone of cell division formed, within the zone of hypertrophy, such that labeled cells appeared to be set among chondrocytes with hypertrophic morphology. At stages after 4 weeks of age the focal feature disappeared as the growth plate returned to more normal morphology by maturity. It seems that this unique "second" zone of dividing cells may contribute to formation of an elongation of the nonresorbed tongue of cartilage. However, it is not likely to be the primary defect since growth plate changes were apparent at earlier stages.

Animals

Immune complexes in ankylosing spondylitis.

Immune complexes have been reported in ankylosing spondylitis (AS) and may implicate infectious agents. Serum samples from 49 patients with AS were assayed for immune complexes by polyethylene glycol precipitation, followed by radial immunodiffusion and pepsinogen binding immunoassay. Both methods showed increases in IgA containing immune complexes, which correlated with serum IgA and with IgA rheumatoid factor concentrations, but did not show increases in other immune complex components. Increased immune complexes were associated with peripheral joint synovitis, but showed no correlation with other clinical or laboratory indices of disease activity. Immune complexes from nine AS serum samples and one AS synovial fluid were electrophoretically separated then probed with anti-Klebsiella pneumoniae, but AS specific antigens were not identified. This study did not suggest a major role for immune complexes in AS without peripheral disease, nor provide serological evidence for the involvement of klebsiella antigens.

Antigen-Antibody Complex

Absence of a specific effect of free radicals on HLA-B27.

The spondylitis associated HLA-B27 epitope includes a characteristic unpaired cysteine at amino acid position 67. On some B27 molecules the thiol (-SH) side chain of this residue seems to be available for chemical interactions. The possibility that free radicals produced during inflammation might specifically affect this group was investigated in this work. Cells bearing HLA-B27 were exposed to free radicals generated by ultraviolet irradiation or hydrogen peroxide, and HLA antigens were then measured by flow cytometry. Binding of monoclonal antibodies to B27 was not affected. These results do not support a specific susceptibility of HLA-B27 to damage by free radicals, despite its apparently vulnerable structure.

4-Chloromercuribenzenesulfonate

HLA-B27 subtypes.

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Epitopes

Tich: a mutant causing disproportional growth in the mouse.

A spontaneous mutation 'tich' (gene symbol tch) appeared as a recessive mutation in inbred mice of strain A. TL. Homozygotes are rather dumpy mice of approximately normal weight but with short limbs and tail. Skeletal measurements on backcross siblings show that the mandible bones are almost normal but long bones and some parts of the pelvic and pectoral girdles are short. Although tich resembles brachypodism phenotypically it is not linked to agouti, and does not match the description of any other skeletal mutation. There was some evidence for weak linkage with albinism on chromosome 7. The mutation has reappeared amongst the A. TL mice of a UK commercial breeder and may have been accepted as the norm for A. TL amongst some European users of this mouse.

Animals

Specific antibody response to the mycobacterial 65 kDa stress protein in ankylosing spondylitis and rheumatoid arthritis.

Immune responses to conserved, immunogenic homologues of the mycobacterial 65 kDa stress protein (SP65) have been implicated in inflammatory arthritis. Serum anti-SP65 was measured in AS, RA and healthy controls using an indirect enzyme immunoassay with recombinant SP65. IgA anti-SP65 was elevated in 19 of 59 AS patients, but the elevation in median level was not statistically significant. Anti-SP65 of all isotypes was increased in RA, but achieved significance (P less than 0.01) for IgA only. Adjusting specific antibody results for elevations in total serum Ig levels reduced AS and RA anti-SP65 to near normal levels, suggesting that a major component of the increased anti-SP65 may be secondary to polyclonal activation.

Antibodies

Effect of a free sulphydryl group on expression of HLA-B27 specificity.

Sequence studies indicate that the alpha-1 domain of the HLA-B27 molecule has a characteristic unpaired cysteine residue at position 67, adjacent, because of secondary structure, to a lysine at position 70. Simple chemical considerations predict that this cysteine should have an exceptionally reactive sulphydryl group. We have shown by ELISA and flow cytometry that the binding of some monoclonal antibodies to B27 on lymphoid cell lines can be inhibited by reagents which react with sulphydryl groups. However this inhibition is never complete: the evidence suggests 2 forms of B27 molecule, one of which is already blocked. We propose that some HLA molecules with oxidised sulphydryls are recognised as different from the reduced forms. Whether they are also recognised as foreign will depend on an individual's history of thymic learning. Oxidation to 'foreign' HLA in the adult is likely to predispose to inflammatory reactions.

Antibodies, Monoclonal

Attempt to show inhibition by oxygen in the modifying factor assay.

An Australian laboratory has demonstrated that antibodies to certain strains of enteric bacteria react with cells from HLA-B27-positive patients with a 'modifying factor' (MF) from these bacteria. These observations have proved difficult to reproduce. We investigated the possibility that oxygen inhibits the test by blocking B27, but we were unable to obtain consistent positive results.

Antibodies, Bacterial

Binding of monomeric and aggregated immunoglobulin to enzymes. A source of artefact in antibody assays.

Pepsinogen has previously been shown to bind non-specifically to immune complexes and aggregated immunoglobulins. We demonstrate here using a solid-phase immunoassay that immunoglobulins aggregated by heat or glutaraldehyde bind non-specifically to several different enzymes. Some of these, including pepsinogen (marketed as pepsin), hyaluronidase and trypsin, are used in the breakdown of tissues or biochemical preparations during the preparation of antigens. Contamination of impure antigens by enzyme is likely to lead to products which bind non-specifically to immune complexes. This can cause misidentification of complexes as antibodies. We recommend that all tests for specific antibody involving the use of antigens prepared by these or other enzymes should include a control with aggregated immunoglobulin substituted for the test serum.

Antibodies

Status of an unpaired thiol group on the HLA-B27 epitope.

Sequence analysis and site-directed mutagenesis of HLA-B27 indicate an unpaired cysteine at position 67 of the hypervariable region corresponding to its serologically defined, disease-associated epitope. We investigated whether chemical modification of this thiol group affected the serological reactions of B27. B27-positive cells were treated with thiol-blocking agents and then tested for recognizable B27 expression. Anti-HLA-B27 alloantisera and monoclonal antibodies were used in cytotoxicity, absorption, and cellular ELISA (cELISA). The semi-quantitative cytotoxicity-based assays showed some decrease in both B27 and controls. However, cELISA indicated that the inhibition of B27 was significantly greater than control antigens, and dependent on thiol-blocker concentration. This suggests that a proportion of HLA-B27 molecules have a free, reactive thiol at the antibody-defined epitope. Incomplete inhibition by thiol-blocking agents indicates that the remainder are inaccessible.

Antibodies, Monoclonal

Biomarkers of aging: tissue markers. Future research needs, strategies, directions and priorities.

Objective tests that allow early detection of deleterious changes with age are necessary to develop treatments enhancing the health span--the length of healthy life. Here we report tests of eight biological systems that can be performed in mice with no harm to the subjects. Male and female B6, CBA and F1 mice were used. While most test results correlated with chronological age in most genotypes, none predicted subsequent longevities in more than two genotypes. Surprisingly, the open field activity test that most consistently predicted longevities, did not correlate with chronological age. Six tests predicted beneficial effects of food restriction in F1 males, but only one correctly predicted the deleterious effects of the same food restriction regimen in B6 males. These results suggest that different biological systems age at different rates, that rates are affected by genotype and that an anti-aging treatment beneficial in one genotype may be harmful in another.

Aging

The role of HLA-B27 in arthritis.

We postulate (Table I) that ReA is an antigen or immune complex induced condition caused by chronic intracellular bacterial infection at a distant site. The main predisposing factor is a failure to resist this infection. If the bacteria happen to carry MF, the inflammation is exacerbated in B27 positive patients. In contrast AS occurs in individuals who lack immunity to MF and eventually become infected by an intracellular organism which synthesizes it (virus or plasmid?). HLA-B27 acts only at the site of inflammation.

Arthritis

Characterization of an unclassified microaerophilic bacterium associated with gastroenteritis.

Four isolates of an unclassified microaerophilic bacterium resembling Campylobacter species were characterized by growth requirements, microscopic examination, biochemical characteristics, antimicrobial susceptibility tests, and protein profile analysis. The unclassified isolates were differentiated from Campylobacter jejuni, Campylobacter coli, Campylobacter fetus subsp. fetus, Campylobacter laridis, Campylobacter pylori, and an ovine isolate. The bacterium was fusiform shaped with a corrugated surface due to the presence of periplasmic fibers and had multiple bipolar flagella. Biochemically, the bacterium was separated from the Campylobacter controls by its negative catalase reaction, negative nitrate reduction, and no growth in 1% glycine. It was also resistant to ampicillin. Protein profile analysis demonstrated nine major protein bands present in the unclassified isolates that were absent in the Campylobacter controls. The bacterium also differed from the ovine isolate by its negative catalase reaction, rapid urea hydrolysis, and susceptibility to clindamycin, erythromycin, and tetracycline. Our results showed that the unclassified bacterium was distinct from the recognized Campylobacter species.

Animals

Case report of an unclassified microaerophilic bacterium associated with gastroenteritis.

An unusual microaerophilic gram-negative bacterium was isolated from the stools of two individuals presenting with chronic diarrhea. This bacterium resembled Campylobacter species by colonial morphology and biochemical reactions. However, microscopic examination revealed a fusiform rod with a corrugated surface, rather than a spiral rod. This is the first reported isolation of this bacterium from humans.

Adult

Epidemiologic study of canine blastomycosis in Wisconsin.

An epidemiologic study was designed to investigate the increasing number of cases of canine blastomycosis being reported in Wisconsin. From January 1980 through July 1982, 200 cases of canine blastomycosis from 39 Wisconsin counties were examined to assess epidemiologic and environmental aspects of this disease. Based on a survey of 176 dog owners, principal disease characteristics for canine blastomycosis were anorexia, lethargy, shortness of breath, chronic cough, and weight loss. The greatest number of cases of canine blastomycosis was in the northwest, north central, northeast, central, and southeast regions of Wisconsin. The northeast and central regions were determined to be new enzootic areas. Sporting breeds accounted for the largest percentage of cases among the various breeds of dogs in Wisconsin. Most of the affected dogs were 3 years old or younger and there was no apparent sexual predilection. Canine blastomycosis was diagnosed more frequently from late spring through late fall. Enzootic areas, except for the southeast region of Wisconsin, were located where the soil was sandy and acid. The results of this study suggested a possible association of enzootic areas with waterways, especially impoundments.

Animals