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Biomedical subjects

M F Lavin

Publications and source records attributed to M F Lavin.

At least 181 records · Page 10Linked to original sources
PubMed1987

Coupling of histone mRNA levels to radioresistant DNA synthesis in ataxia-telangiectasia cells.

Cloned genomic DNA for human histone H1, H3 and H4 genes has been used to determine the effects of gamma-radiation on histone mRNA levels and synthesis in ataxia-telangiectasia cells. Synthesis of histone mRNA was determined in cells synchronized with aphidicolin. Effects of irradiation on DNA synthesis and passage through S phase were also monitored. Irradiation was found to slow the passage of control cells through the cell cycle but had no effect on progression of ataxia-telangiectasia cells. H1 and core histone mRNA synthesis was inhibited by radiation in two control cell lines after release from aphidicolin block. No inhibition was observed in one ataxia-telangiectasia cell line and a small degree of inhibition in a second. An increased level of mRNA was observed in both irradiated control and ataxia-telangiectasia cells at 5-7 h post-irradiation compared to unirradiated cells. Similar results were obtained in log phase cells. These results demonstrate that histone mRNA synthesis is radioresistant in ataxia-telangiectasia cells and is coupled to radioresistant DNA synthesis in these cells.

Ataxia Telangiectasia↗
PubMed1987

Transferrin receptor expression by human bladder transitional cell carcinomas.

The expression of transferrin receptors (TFR) by normal and neoplastic urothelial cells was studied in "control" patients and in patients with transitional cell carcinoma of the bladder. These tumours were graded independently and consisted of 19 grade I, 30 grade II and 19 grade III lesions. TFRs were identified using a monoclonal antibody specific for TFR (OKT9) in an immunofluorescent or avidin/biotin-immunoperoxidase technique on fresh frozen sections. TFRs were not detected on normal urothelium. However, positive staining was found to increase with increasing pathological grade and stage of the tumours, ranging from 31.6% of grade I to 78.9% of grade III tumours and 51.2% of pTa (mucosa only lesions) to 87.5% of pT2/pT2+ (muscle invasion +/- deeper) primary urothelial malignancies.

Adult↗
PubMed1987

Genotoxicity of analgesic compounds assessed by an in vitro micronucleus assay.

Several analgesic compounds and mixtures of analgesics were examined for both cytotoxicity and ability to induce chromosomal damage in the normal rat-kidney cell line NRK-49F. Chromosomal damage was assessed using an in vitro micronucleus assay. Of all the compounds tested, only N-hydroxyparacetamol caused a high degree of cell death at the concentrations used. 4 analgesic compounds were found to be inducers of micronuclei in NRK cells; in order of decreasing potency these were: N-hydroxyparacetamol, N-hydroxyphenacetin, caffeine and paracetamol. An aspirin, phenacetin, caffeine mixture (APC) failed to induce micronuclei above the background level, and a paracetamol-codeine combination did not increase the level of micronuclei induction above that induced by paracetamol alone. This report suggests paracetamol and some related compounds are capable of inducing chromosomal damage in mammalian cells in vitro, which is consistent with recent reports of a possible paracetamol-DNA interaction.

Analgesics↗
PubMed1987

Inhibition of DNA, RNA and protein synthesis and chromatin alteration by N-hydroxyphenacetin.

The effects of N-hydroxyphenacetin on DNA function and structure were investigated to elucidate the involvement of phenacetin in analgesic nephropathy and transitional cell carcinoma. N-Hydroxyphenacetin or a metabolite inhibited synthesis of DNA, RNA and protein; DNA inhibition was greater at higher pH. No single-strand breaks were detectable in DNA after N-hydroxyphenacetin treatment and no appreciable effect on cell viability was observed at concentrations up to 5 mM. N-Hydroxyphenacetin-induced alteration to chromatin structure was detected using nucleoid sedimentation analysis. Direct binding to plasmid DNA was not observed. These observations are consistent with a role for phenacetin metabolites in renal disease.

Cell Line↗
PubMed1987

DNA fragmentation in childhood T-cell acute lymphoblastic leukaemia.

DNA isolated from 12 patients with childhood acute lymphoblastic leukaemia (T-ALL), analysed by gel electrophoresis exhibited a degradation pattern of increasing nucleosome repeat size, similar to that observed when nuclei from normal control cells are incubated with micrococcal nuclease. None of a group of adult T-ALL showed this fragmentation, nor was the pattern observed in normal control T-cells, cells from patients with a number of other forms of leukaemia, or in B-lymphoblastoid cells under the same incubation conditions. This phenomenon appears to be related to the changes in DNA that occur in glucocorticoid-induced cell death in mouse thymocytes, radiation-damaged spleen cells, and in cytotoxic T-cells deprived of interleukin 2. The method described here promises to be a reliable, simple approach to the characterization of T-ALL in childhood and may aid in the clinical management of this form of leukaemia.

Adolescent↗
PubMed1987

Bovine ocular squamous cell carcinoma: superoxide dismutase and catalase levels.

Considerable evidence exists for the production of free radicals in the eye region after exposure to ultraviolet light. Failure to remove these radicals adequately may be important in the predisposition of Hereford cattle to ocular carcinoma. The levels of two enzymes, superoxide dismutase and catalase, involved in the removal of superoxide anion and hydrogen peroxide, were compared in Hereford cattle predisposed to ocular carcinoma and a resistant breed, Droughtmaster cattle. The results demonstrate comparable levels of superoxide dismutase in the two breeds in both eye tissue and erythrocytes. Catalase levels also were the same in both breeds. Increased incidence of ocular carcinoma in Herefords does not appear to be due to a deficiency in either superoxide dismutase or catalase.

Animals↗
PubMed1986

Isolation of 3,4-dihydroxyphenylalanine-containing proteins using boronate affinity chromatography.

A rapid procedure for the isolation of 3,4-dihydroxyphenylalanine-containing proteins has been developed in which the protein is selectively bound to a m-phenylboronate agarose column, and eluted with 1.0 M ammonium acetate, pH 3.0. The method is based on the affinity of boronates for diols including catechol. The chromatography is carried out in the absence of oxygen to prevent oxidation of the catechol. Other proteins are eluted beforehand with 0.25 M ammonium acetate, pH 8.5, or for glycoproteins with a Tris buffer containing 0.2 M sorbitol, pH 8.5.

Animals↗
PubMed1986

Effect of N-hydroxyparacetamol on cell cycle progression.

N-hydroxyparacetamol treatment of rat kidney cells gave rise to a dose-dependent decrease in DNA synthesis. A concentration of 1.0 mM N-hydroxyparacetamol at pH 7.2 decreased the level of DNA synthesis to 13.0 +/- 2.3% of the control value after 1 hr incubation. This compound also caused a perturbation of cell cycle progression. A concentration of 0.44 mM N-hydroxyparacetamol induced G1/S and S phase blocks. These delays became evident at approximately 12 hr after treatment and persisted until about 15 hr when cells started to recover. It seems unlikely that N-hydroxyparacetamol inhibits DNA synthesis and perturbs cycle progression through alterations to DNA structure as such, since this compound failed to alter the migration pattern of naked plasmid DNA.

Acetaminophen↗
PubMed1986

Study of chromatin structure in ataxia-telangiectasia cells.

Micrococcal nuclease was used as a probe to study chromatin structure in control and ataxia-telangiectasia cells. The rate and extent of release of acid-soluble nucleotide was similar in both cell types. Production of mono- and oligonucleosomes by micrococcal nuclease as determined by gel electrophoresis also failed to reveal differences in chromatin structure between control and ataxia-telangiectasia cells. Radiation exposure did not significantly alter the kinetics of digestion. These results indicate that there are no gross alterations in chromatin structure in ataxia-telangiectasia cells.

Ataxia Telangiectasia↗
PubMed1986

Ataxia-telangiectasia cell extracts confer radioresistant DNA synthesis on control cells.

We have investigated in greater detail the radioresistant DNA synthesis universally observed in cells from patients with ataxia-telangiectasia (A-T). The approach employed in this study was to permeabilize cells with lysolecithin after gamma-irradiation and thus facilitate the introduction of cell extract into these cells. This permeabilization can be reversed by diluting the cells in growth medium. Cells treated in this way show the characteristic inhibition (control cells) or lack of it (A-T cells) after exposure to ionizing radiation. Introduction of A-T cells extracts into control cells prevented the radiation-induced inhibition of DNA synthesis normally observed in these cells. A-T cell extracts did not change the level of radioresistant DNA synthesis in A-T cells. Control cell extracts on the other hand did not influence the pattern of inhibition of DNA synthesis in either cell type. It seems likely that the agent involved is a protein because of its heat lability and sensitivity to trypsin digestion. It has a molecular weight (MW) in the range 20-30 000 D. The development of this assay system for a factor conferring radioresistant DNA synthesis on control cells provides a means of purifying this factor, and ultimately an approach to identifying the gene responsible.

Ataxia Telangiectasia↗
PubMed1986

Rearrangements of T-cell receptor beta-chain genes in human leukaemias.

Rearrangements of the T-cell receptor (TCR), beta-chain genes and immunoglobulin (Ig) heavy chain genes in several T-cell leukaemias (T-ALL and ATL), and some B-cell and myelogenous leukaemias were investigated. Two out of 15 cases of T-cell leukaemia tested failed to show a rearrangement pattern of TCR beta genes although both expressed mRNA for this gene. The remaining 13 cases showed diverse patterns of rearrangements involving either C beta 1, C beta 2 or both. C beta 1 but not C beta 2 was deleted in some of the T-cell leukaemias. Polyclonal T cells from four normal individuals showed the germ line pattern and an additional two bands in Hind III digested DNA. Except for one, all cases of C-ALL (B-cell leukaemia) showed a rearranged JH locus which was not evident in any of T-cell leukaemias studied. One case of B-cell leukaemia showed a rearrangement of both TCR beta genes and JH genes. The results of these studies suggest that rearrangement of TCR and Ig genes occurs at a very early stage of differentiation of stem cells and does not appear to play a direct role in leukaemogenesis per se.

Antigens, Neoplasm↗
PubMed1986

Differential distribution of ATPase- and T6-positive cells (Langerhans cells) in the limbus and cornea of Hereford and non-Hereford cattle.

Epithelial sheets from the limbus, cornea, and third eyelid of Hereford and non-Hereford cattle were examined for the presence of Langerhans cells (LC) using the membrane enzyme ATPase as a marker for LC. The aim of the study was to test the hypothesis that differences in LC density exist between the various ocular epithelia of these animals producing depressed immune surveillance in the case of Hereford cattle. The presence of LC in ocular tissues was confirmed by parallel studies which detected epithelial cells bearing T6, an antigen expressed by human LC. Studies using serial sections demonstrated that T6+ cells also reacted with an anti-human HLA-DR monoclonal antibody. The detection of T6+, DR+ and ATPase+ cells in ocular epithelium in the absence of infiltrating macrophages suggested that LC are present in these tissues. While there were no significant differences in the density of T6+ cells between non-Hereford and Hereford cattle, in the latter ATPase+ cells were significantly fewer in the lateral, medial, and upper limbus.

Adenosine Triphosphatases↗
PubMed1986

Ionizing radiation and DNA-chain elongation in ataxia telangiectasia lymphoblastoid cells.

DNA-chain elongation rates, determined by sedimentation analysis, were found to be similar in control and ataxia-telangiectasia lymphoblastoid cells. A gamma-radiation dose of 6 Gray, which had previously been shown to have a marked inhibitory effect on initiation of DNA replication, had no appreciable effect on elongation rates in either cell type. Elongation rates were also determined at 20 Gray of gamma-rays by pulsing cells with [3H]thymidine prior to irradiation to avoid anomalous sedimentation behaviour. At this radiation dose elongation was almost completely inhibited in control cells while little or no inhibition was observed in ataxia-telangiectasia cells. Deoxyribonucleoside triphosphate pool equilibration times were not altered at either dose.

Ataxia Telangiectasia↗
PubMed1986

Immunohistochemical analysis of the human bladder.

An immunohistochemical profile of cells in the mucosa and lamina propria of adult human bladders is described. Urothelial cells were HLA-DR-ve, ACP + ve and ATPase-ve. All lymphocytes in this layer were of the suppressor/cytotoxic T subgroup and dendritic cells with an identical phenotype to Langerhans cells were seen. In the lamina propria most lymphocytes were of the suppressor/cytotoxic type with some helper/inducer cells. An occasional natural killer cell and Langerhans cell was identified and large macrophage cells with a common phenotype to antigen presenting interdigitating (ID) cells were noted. Fibronectin staining was dense in the vicinity of basement membranes merging to form fine interconnecting latticework-like structures elsewhere in the lamina propria.

Acid Phosphatase↗
PubMed1986

Effect of N-hydroxyparacetamol on DNA, RNA, and protein synthesis and chromatin structure.

N-Hydroxyparacetamol treatment of lymphoblastoid cells gave rise to a dose-dependent decrease in DNA, RNA, and protein synthesis. Inhibition of DNA synthesis was less marked in medium at pH 6.5 than at pH 9.0. N-Hydroxyparacetamol appeared to inhibit DNA synthesis at least in part through alterations to chromatin structure. This compound produced a dose-dependent and time-dependent loss in the superhelix density of DNA as determined by nucleoid sedimentation analysis. Alkaline elution data as well as sucrose gradient analysis revealed that this decrease in sedimentation did not arise through single strand breakage to DNA. The structural alterations to chromatin caused by N-hydroxyparacetamol appeared to have been repaired after 6 hr. However, sedimentation of "repaired" nucleoids in the presence of ethidium bromide was markedly different from sedimentation of untreated nucleoids. These results suggested that some N-hydroxyparacetamol remained associated with nucleo-protein, thus interfering with the binding of ethidium. Furthermore, both RNA and protein synthesis were markedly inhibited by N-hydroxyparacetamol, demonstrating a major effect on cell function. The widespread effects of N-hydroxyparacetamol could be accounted for by changes to chromatin structure or by a more general effect on cellular metabolism. Either of these effects could account for the dramatic cytotoxicity of this compound. A concentration of 2.5 mM reduced cell viability by 96% after 3 days.

Acetaminophen↗
PubMed1985

p-Aminophenol induced DNA damage and cytotoxicity enhanced by autoxidation.

p-Aminophenol inhibits DNA synthesis and alters the structure of DNA. A decrease in sedimentation of nucleoids from cells treated with p-aminophenol was observed and this decrease in sedimentation was considerably less when cells were incubated with p-aminophenol in an atmosphere of nitrogen or at lower pH values. This compound was also shown to be cytotoxic to cells in culture. These results demonstrate that conditions retarding the autoxidation of p-aminophenol lead to reduced effects on DNA structure and a lesser cytotoxic effect.

Aminophenols↗
PubMed1985

Effect of caffeine on gamma-ray-induced G2 delay in ataxia telangiectasia.

Exposure of normal control and ataxia-telangiectasia (A-T) lymphoblastoid cell lines to ionizing radiation gives rise to an increase in the proportion of G2 phase cells. The size and extent of the G2 phase block is greater in A-T cells than in normal cells. Caffeine has a similar overall effect in control and A-T cell lines in reducing the G2 arrest observed after ionizing radiation. While the proportion of cells accumulated in G2 in A-T cells is considerably greater than in controls, addition of caffeine at the time of maximal G2 block brings about a return of G2 phase cell numbers to unirradiated values in 3 hours in both cell types. In normal control cells the caffeine-mediated decrease in G2 cells is reflected by an increase in mitotic cells. These mitotic cells have a higher frequency of chromosome aberrations compared to cells harvested in the absence of caffeine. Similarly in A-T cells addition of caffeine to irradiated cultures, delayed in G2 phase, increased the number of mitotic cells and the frequency of chromosome aberrations.

Ataxia Telangiectasia↗