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Biomedical subjects

D I Hoar

Publications and source records attributed to D I Hoar.

At least 55 records · Page 3Linked to original sources
PubMed1986

T lymphocytes from hemophiliacs proliferate after exposure to factor VIII product.

Chronic exposure of hemophiliacs to allogeneic proteins via factor VIII concentrate for control of bleeding could lead to a state of chronic antigen stimulation. Immune function from eight hemophiliacs requiring large amounts of factor VIII concentrate was assessed. It was found that 5 of 8 had a positive blastogenic response to low concentrations of factor VIII in a 7-day thymidine uptake assay. Separation of lymphocyte subpopulations indicated that the reactivity was contained in the T cell-enriched fraction. There was no blast transformation noted secondary to the factor VIII product in 20 controls tested in a 7-day assay nor was there any mitogenic effect of the factor VIII in a 3-day assay. In fact, high concentrations of factor VIII impaired that T lymphocytes from hemophiliacs are antigenically primed to some constituent in the lyophilized factor VIII concentrate.

Adult↗
PubMed1986

Ground squirrel hepatitis virus (GSHV) infection and hepatocellular carcinoma in the Canadian Richardson ground squirrel (Spermophilus richardsonii).

Sera and livers from 40 Richardson ground squirrels were examined for evidence of ground squirrel hepatitis virus (GSHV) infection and hepatocellular carcinoma. Twenty-five sera were obtained from fully grown adult ground squirrels and 15 from young ground squirrels estimated to be between 6-8 weeks of age. All animals had been caught in the wild and had spent less than 1 month in captivity. Sixteen sera (40%) had at least one serologic marker of GSHV (2 with GSHV surface antigen, 3 GSHV core antigen, 5 GSHV antibody to core antigen and 11 GSHV antibody to surface antigen). Two cases of hepatocellular carcinoma were detected, both in adult ground squirrels. Tumour tissue and adjacent normal liver tissue were negative for GSHV surface and core antigen by direct immunofluorescence in both livers and negative for GSHV-DNA by molecular hybridization in the one tumour examined. These results indicate that: A) GSHV is geographically more widespread than previously considered; B) viral transmission occurs early in life, probably via the vertical route; and C) hepatocellular carcinoma is a relatively common finding in these animals while still in their wild state. Any causal relationship between hepatocellular carcinoma and GSHV infection in these animals, however, has yet to be demonstrated.

Animals↗
PubMed1986

Natural killer cell activity from hemophiliacs exhibits differential responses to various forms of interferon.

Patients with hemophilia are at risk for the development of acquired immunodeficiency syndrome (AIDS). Patients with AIDS have recurrent infections and/or malignancy and altered immune response, including decreased T lymphocyte counts, decreased T helper lymphocytes, defective T cell blastogenesis, hypergammaglobulinemia, defective natural killer (NK) activity and impaired response of NK to interferon-beta (IFN-beta). It is feasible that chronic antigen stimulation with subsequent release of interferon could be related to the impaired NK reactivity to IFN-beta of patients with AIDS. Because hemophiliacs are subjected to chronic antigen stimulation secondary to the administration of foreign protein, the reactivity of NK cells from patients with hemophilia to IFN-alpha, IFN-beta and IFN-gamma was studied. Eight patients with hemophilia requiring high levels of clotting factor replacement were assessed. Three patients were antibody positive to HTLV-III. All had normal baseline NK cell function. In the first set of experiments, all patients responded normally to in vitro IFN-alpha by increasing NK activity, but four patients had significant failure and two had mild impairment in NK response to IFN-beta. This latter observation was particularly evident at very low concentrations of IFN. In repeated experiments, seven of eight had impaired NK response to IFN-beta and IFN-gamma but normal response to IFN-alpha. Only one patient's NK cells responded better to IFN-gamma. There was no obvious correlation of these findings to antibody status to HTLV-III. Chronic antigen stimulation and the modulation of interferon receptors are discussed as possible mechanisms that could produce these findings.

Adult↗
PubMed1985

Induction of mitochondrial mutations in human cells by methotrexate.

Inhibition of dihydrofolate reductase by the folate analog, methotrexate (MTX) results in a depletion of tetrahydrofolate dependent one carbon transfer reactions in amino acid and nucleic acid biosynthesis. When human cells (either HeLa or normal skin fibroblasts) are exposed to MTX in a defined medium containing dialyzed fetal calf serum, essential and non-essential amino acids, and purine source, the thymidylate pools alone are depleted. Under these conditions exposure to 10(-6) M MTX induces mitochondrial mutagenesis, measured as an increase in the frequency of chloramphenicol resistant (CAPR) colonies, without altering the rate of nuclear mutation monitored by determining the frequency of 6-thioguanine resistance (TGr). The occurrence of CAPR mutations is time, and MTX concentration dependent and the frequency of CAPR can be decreased quantitatively by adding thymidine to the culture medium. This mitochondrial specific mutagenesis can also be achieved using the thymidylate synthetase inhibitor, 5-fluorodeoxyuridine further implicating thymidylate pools as the mediator of this effect. During the course of exposure to 10(-6) M MTX the thymidine kinase deficient HeLa BU25 cell line exhibits a progressive depletion and degradation of mitochondrial DNA suggesting that the mutagenesis and DNA degradation represent portions of a progressive process. The basis for the selective sensitivity of the mitochondrial genome to thymidylate depletion mutagenesis may be the consequence of its differences from the nuclear genome in mechanisms of DNA replication or repair.

Animals↗
PubMed1985

Hepatitis B virus DNA is enriched in polymorphonuclear leukocytes.

DNA hybridization and cell separation techniques were used to determine which blood components contained hepatitis B viral DNA sequences. Free monomer-length hepatitis B virus was found in large amounts in the polymorphonuclear leukocyte cell fraction in two of five HBsAG-positive patients. In these two patients, viral DNA sequences were not detected in the plasma or platelet fraction, whereas the mononuclear cell DNA contained small amounts of a 7.2 kb size unintegrated hepatitis B genome. These studies indicate that the major reservoir of unit-length viral DNA in the asymptomatic hepatitis B carriers studied here was in the polymorphonuclear leukocyte fraction. The basis for the presence of the viral DNA within these cells is presently unknown, but may relate to viral replication within, or phagocytosis of virus by, these cells.

Carrier State↗
PubMed1984

Factors distinguishing couples at risk for nondisjunction.

Micronucleus frequencies were determined on 24 young parents of trisomic infants, 21 individuals with recurrent unexplained abortions, and 42 control individuals with proven reproductive success. In addition to measurements of spontaneous micronucleus frequencies, mitomycin C induced frequencies were determined at two doses (2.5 and 5.0 ng/mL). Using the 95% confidence limits established from control data as an arbitrary upper limit, 16 of 24 parents of trisomics and 5 of 21 recurrent aborters were detected by the micronucleus assay to above this cutoff. The effects of sex, age, pregnancy status, and a variety of environmental exposures were studied by comparing the micronucleus frequencies of the exposed and unexposed populations. The data suggested that vitamins were associated with a lower micronucleus frequency and tea drinking with an increased micronucleus frequency in parents of trisomics, an effect not seen in controls. These results suggest that a biologic basis for nondisjunction may be associated with elevation in spontaneous and induced micronuclei. In addition, tea and vitamins may modulate micronucleus frequencies in parents of trisomics who appear to be more sensitive to these influences than the controls.

Abortion, Habitual↗
PubMed1984

A frequent restriction fragment length polymorphism in the human metallothionein-II processed gene region is evolutionarily conserved.

Genomic blot analysis of human DNA indicated that metallothioneins are represented by a multi-gene family. Clones containing metallothionein sequences have been isolated and two of these have been identified as metallothionein-I and metallothionein-II processed genes by sequence analysis. The metallothionein-II processed gene in humans shows two restriction fragment length polymorphisms of 4.5 and 4.8 kb (10(3) bases) when EcoRI-digested genomic DNA from various individuals was analysed by Southern blotting. All the three genotypes are found at a high frequency and thus the metallothionein-II processed gene represents a true polymorphic marker. Familial studies also indicate that these restriction fragment length polymorphisms follow the classical Mendelian inheritance. Detailed Southern blot analyses show that this restriction fragment length polymorphism is due to a restriction site polymorphism and is localized at the 5'-flanking region of the metallothionein-II processed gene. Sequence analysis of the suspected region in the 4.8 kb fragment shows that the sequence G*GATTC, which is found 371 nucleotides downstream from the EcoRI site on the 5' end of the 4.8 kb fragment, makes a HinfI site. A transition of *G to A in this sequence in the 4.5 kb allele has resulted in loss of the HinfI site and created an EcoRI site. Thus, this mutation has given rise to this restriction fragment length polymorphism.

Alleles↗
PubMed1979

Host-cell reactivation of UV-irradiated adenovirus in Cockayne syndrome.

Measurements of the host-cell reactivation (HCR) of mutagen-treated virus provides a very sensitive tool for detecting abnormal DNA repair. The best example of the utility of HCR studies in the examination of the DNA-repair capacity of human cells has come from studies of cells from the UV-sensitive repair-deficient xeroderma pigmentosum (XP) patients. We have examined the HCR of UV-treated adenovirus type 5 (Ad5) and type 2 (Ad2) in cells from patients with Cocayne syndrome (CS), another sun-sensitive syndrome whose cells also exhibits UV-sensitivity in culture. Comparisons with obligate heterozygotes and normal controls failed to reveal an abnromality in the HCR capacity of the CS cells. As the abnormality in DNA metabolism in CS appears to be in a late step in excision repair, a bypass mechanism may exist in these cells for circumventing the defect in the repair of viral DNA.

Adenoviridae↗
PubMed1979

Familial mutagen sensitivities: are these the hallmarks of meiotic or mutator mutants in humans?

Mutagen hypersensitivity (MHS) has been found to be associated with abnormalities in DNA metabolic processes in many prokaryotic and eukaryotic organisms. The study of fibroblasts derived from humans with genetic diseases believed to have altered DNA metabolism, has also revealed patterns of MHS. In this paper results are presented that suggest MHS patterns unrelated to obvious disease can be inherited in a dominant fashion. As these individuals exhibiting MHS patterns have been observed in families with poor reproductive history, new syndromes, or ontogenetic problems (including malignancies) there may be a causal relationship between these events and the MHS. These events which may have a genetic basis appear as maternal or paternal effect mutants as the consequences are observed in reduced reproductive fitness or abnormal progeny. Since these effects are similar to the events precipitated by the meiotic or mutator mutants in Drosophila, it is speculated that the MHS patterns may be the hallmarks of such mutants in man.

Ataxia Telangiectasia↗
PubMed1978

DNA repair in Cockayne syndrome.

Cockayne syndrome (CS) is a rare recessive genetic disease characterized in part by premature ageing and photosensitive skin. Because of the latter characteristic, this syndrome was considered to be an example of a UV-sensitive DNA repair-defective human disorder. We demonstrated normal levels of UV-induced unscheduled DNA synthesis (UDS) in four unrelated CS patients that show hypersensitivity to both UV and Mitomycin C (MMC). At low UV exposure, CS DNA shows a dose-dependent decrease in size. By contrast, heterozygotes appear to have a threshold below which there is little change in size of single strand DNA. Immediately following UV or MMC treatment, CS DNA is deficient in high molecular weight species, but undergoes a normal transition to larger DNA during a chase interval in the presence or absence of caffeine. This suggests a defect in replication or excision repair and no defect in post-replication repair (PRR). Pulse studies performed in the presence of hydroxyurea (HU) also reveal a deficient production of large DNA, suggesting the defect is in repair. As these cells have normal UDS and normal PRR, the basis for their UV sensitivity must be distinct from that observed in xeroderma pigmentosum (XP).

Cells, Cultured↗