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Biomedical subjects

T Burke

Publications and source records attributed to T Burke.

At least 19 recordsLinked to original sources

Multilocus DNA fingerprints in gallinaceous birds: general approach and problems.

Multilocus profiles were investigated in five different species of Galliformes (ring-necked pheasant Phasianus colchicus, Indian peafowl Pavo cristatus, Japanese quail Coturnix coturnix japonica, domestic chicken Gallus gallus, and red grouse Lagopus lagopus scoticus) using two human multilocus probes (33.6 and 33.15) in combination with each of four restriction enzymes (AluI, DdeI, HaeIII or HinfI). All the species show a DNA fingerprint-like pattern using at least one restriction enzyme in combination with each multilocus probe. The number of bands detected and the value of the index of similarity for each species differ significantly between the profiles obtained with each multilocus probe. Some enzyme/probe combinations reveal strong cross-hybridization of the multilocus probes with satellite or satellite-like DNA sequences in pheasant, peacock, quail and chicken, which partially or completely prevented scoring of the profile. The choice of restriction enzyme was found to influence the number of bands, the value of the index of similarity and the probability of obtaining an identical fingerprint between unrelated individuals. The Mendelian inheritance and independent segregation of the fragments detected using AluI was investigated in three species (ring-necked pheasant, Indian peafowl and red grouse). Some bands were shown to be tightly linked. An extreme case was encountered in the red grouse, where 12 of the 15 bands scored in one parent represented only two, apparently allelic, haplotypes and so derived from a single locus. However, fingerprint patterns will often be adequate for use in paternity analyses, such as in behavioural studies, despite the occurrence of haplotypic sets of bands. Identical DNA multilocus profiles were sometimes observed between captive-bred siblings in one species. These results emphasize the desirability of determining, in each new species, the optimal experimental conditions as a preliminary to any behavioural or population genetic studies that use the multilocus DNA fingerprinting methodology.

Animals

Human autoantibodies against desmoplakins in paraneoplastic pemphigus.

Recently, a previously unrecognized autoantibody mediated blistering disease, paraneoplastic pemphigus has been described. Paraneoplastic pemphigus is associated with lymphoid malignancies, thymomas, and poorly differentiated sarcomas. Serum of affected patients contain pathogenic autoantibodies that immunoprecipitate from normal keratinocytes a characteristic complex of four polypeptides with M(r) of 250, 230, 210, and 190 kD. As our preliminary studies indicated that the 250-kD and the 210-kD antigens comigrated with desmoplakins I and II, we investigated the possibility that autoantibodies against the desmoplakins were a component of this autoimmune syndrome. 11 sera from affected patients were tested by indirect immunofluorescence against desmosome containing tissues, immunoprecipitation of metabolically labeled keratinocytes, and Western immunoblotting of desmoplakins I and II that had been purified to homogeneity from pig tongue epithelium. By indirect immunofluorescence, 9 of 11 sera showed strong binding to epithelial and nonepithelial desmosomes, and 2 were weakly reactive. All 11 immunoprecipitated 250- and 210-kD bands of variable intensity that comigrated with bands identified by a murine monoclonal antidesmoplakin antibody, and immunoblotting confirmed binding of the serum autoantibodies to purified desmoplakins. This demonstrates that paraneoplastic pemphigus is the first human autoimmune syndrome in which autoantibodies against the desmoplakins are a prominent component of the humoral autoimmune response.

Animals

Multilocus and single locus minisatellite analysis in population biological studies.

In this review we describe the situations in which minisatellite analysis is of value to studies of population and evolutionary biology. Evolutionary and population biologists need to be able to quantify genetic relationships among individual organisms at many different levels, from close familial relationships to evolutionarily distant phylogenetic ones. The use of minisatellite markers is put into this context and compared with the other molecular biological techniques. Examples of the use of multilocus minisatellite analysis in population biology are described. The limitations of multilocus fingerprinting are presented, together with the potential advantages of locus-specific probes. The use of locus-specific probes in population biology is now often feasible due to the recent development of a cloning system which allows their efficient isolation. The availability of locus-specific probes should significantly expand the role of minisatellite markers in population biology.

Animals

Hypervariable minisatellite DNA sequences in the Indian peafowl Pavo cristatus.

We report here for the first time the large-scale isolation of hypervariable minisatellite DNA sequences from a non-human species, the Indian peafowl (Pavo cristatus). A size-selected genomic DNA fraction, rich in hypervariable minisatellites, was cloned into Charomid 9-36. This library was screened using two multilocus hypervariable probes, 33.6 and 33.15 and also, in a "probe-walking" approach, with five of the peafowl minisatellites initially isolated. Forty-eight positively hybridizing clones were characterized and found to originate from 30 different loci, 18 of which were polymorphic. Five of these variable minisatellite loci were studied further. They all showed Mendelian inheritance. The heterozygosities of these loci were relatively low (range 22-78%) in comparison with those of previously cloned human loci, as expected in view of inbreeding in our semicaptive study population. No new length allele mutations were observed in families and the mean mutation rate per locus is low (less than 0.004, 95% confidence maximum). These loci were also investigated by cross-species hybridization in related taxa. The ability of the probes to detect hypervariable sequences in other species within the same avian family was found to vary, from those probes that are species-specific to those that are apparently general to the family. We also illustrate the potential usefulness of these probes for paternity analysis in a study of sexual selection, and discuss the general application of specific hypervariable probes in behavioral and evolutionary studies.

Animals

Chromite ore processing residue in Hudson County, New Jersey.

Chromite ore processing residue occurs at over 130 sites in Hudson County, New Jersey. Many of these sites are in urban residential areas. This waste is a result of 70 years of chromate and bichromate chemical manufacturing. At least 15% of the sites contain total chromium concentrations greater than 10,000 mg/kg, with hexavalent content ranging from about 1 to 50%. Continuing leaching of this waste results in yellow-colored surface water runoff and yellow deposits on the soil surface and inside basement walls. The chemistry, environmental fate, health effects, and human exposure potentials for this waste are described.

Chromium

Doxorubicin resistance conferred by selective enhancement of intracellular glutathione peroxidase or superoxide dismutase content in human MCF-7 breast cancer cells.

To examine the role of doxorubicin-stimulated oxyradical formation in tumor cell killing, we introduced glutathione peroxidase (GSH Px) or superoxide dismutase (SOD) into MCF-7 cells by "scrape loading." Control cytoplasmic GSH Px and SOD levels increased from (mean +/- S.E.) 0.37 nmol/min/mg and 0.58 microgram SOD/mg, respectively, to 3.99 or 7.63 nmol/min/mg and 1.40 or 1.83 micrograms SOD/mg after treatment with either 150 or 300 units/ml of GSH Px or 20 or 40 mg/ml SOD. Resistance to doxorubicin was correlated with the level of GSH Px introduced into the MCF-7 cells; a one-hour exposure to 1.75 microM doxorubicin decreased the cloning efficiency of control cells loaded with medium alone to 34%, whereas doxorubicin-treated cells augmented with 150 or 300 units/ml of GSH Px had plating efficiencies of 56 or 86%, P less than 0.05. Introduction of SOD increased MCF-7 resistance to doxorubicin similarly. The heat-inactivated enzymes were not protective. Cells loaded with GSH Px were also resistant to the redox cycling anticancer quinone mitomycin C but not to the redox inactive analogs 5-iminodaunorubicin and mitoxantrone, suggesting that amplification of GSH Px or SOD levels can produce doxorubicin resistance in MCF-7 cells.

Breast Neoplasms

Proximal femoral replacement for pathological fracture.

Nine patients presented to our unit with extensive metastatic destruction of the proximal femur. The extent of bone loss precluded the use of standard methods of fixation. We undertook proximal femoral replacement as a palliative treatment to restore mobility and relieve pain. Six patients mobilized satisfactorily postoperatively. Three patients did not regain independent mobility and died within 4 weeks of surgery. The major complication was dislocation, which occurred in two patients. The mean postoperative survival was 8 months. We believe proximal femoral replacement is a useful palliative procedure in selected cases of metastatic disease of the femur.

Adult

Drinking water contamination and the incidence of leukemia: an ecologic study.

An ecologic study was performed to examine the relation between the incidence of leukemias and the occurrence of volatile organic chemical (VOC) contamination of drinking water supplies within a study area comprised of subpopulations differentially exposed to drinking water VOCs (trichloroethylene and related solvents). Populations served by community water supplies were classified into exposure categories according to VOC contamination status based on 1984-85 sampling data. Leukemia incidence data (1979-84) were collected from a population-based cancer registry. For females, the standardized incidence ratio was elevated only in towns in the highest of three exposure categories. No association was observed in males in any of the exposure categories. A Poisson regression analysis of the data, using finer exposure strata, indicated an increase in risk among females with increasing level of contamination which appeared to be distributed evenly across all age strata. The rate ratio for females at the highest exposure stratum for total non-THM VOCs compared to the least exposed stratum was 1.68. The observed association appears to suggest that drinking water contaminated with VOCs may increase the incidence of leukemia among exposed females, but caution is advised in the interpretation of these results because of the uncertainties inherent in ecologic studies.

Adult

Sustained-release dosage forms.

Transdermal drug delivery provides an effective and advantageous alternative to the oral delivery of therapeutic agents. Currently, many other drugs are being evaluated for transdermal delivery including testosterone, fentanyl, nicotine and timolol. They offer the promise of simplified dosage regimens, enhanced compliance, reduced side effects and improved therapy of disease.

Administration, Cutaneous

Determinants of diagnostic accuracy in pulmonary scintigraphy for pulmonary capillary protein leak associated with adult respiratory distress syndrome (ARDS): a technical note.

Radionuclide assessment of pulmonary capillary protein leak using [99mTc] human serum albumin (99mTc-HSA) was first reported from our laboratory. In this study we investigated the impact of 1) sampling time post tracer injection, and 2) lung region assignment, on diagnostic accuracy between 2 groups (control n = 20 and ARDS n = 20). Each patient received 370 MBq 99mTc-HSA i.v. and was imaged for 45 min. The slope index (SI) [change in lung: heart activity ratio/min] was calculated from 11 computer assigned lung regions for intervals of 5-15 (early [E]) and 15-45 (late [L]) min. The diagnostic accuracy of E vs L SI calculations for the 11 regions was evaluated by stepwise logistic regression. E SI data and L SI data from the lower 1/3 of the lung did not achieve significance for inclusion in the discriminant model (P less than 0.05). In the nine remaining regions L SI was significant. Optimal discrimination was achieved from L SI data obtained from a region confined to the lateral half of the mid 3rd of the lung field (sensitivity 81%, specificity 85%, accuracy 83%). The results confirm that: 1) a late (15-45 min) sampling period and 2) proper region assignment are necessary to maximize accuracy of this technique.

Adult

Phosphorylation of class I but not class II MHC molecules by membrane-localized protein kinase C.

Membranes were isolated from B cells stimulated with phorbol 12-myristate 13-acetate (PMA) for a time sufficient to allow maximal redistribution and activation of protein kinase C (PKC). Exposure of such membranes to a short incubation with [gamma-32P]ATP resulted in the detection of at least nine unique or hyperphosphorylated membrane proteins by SDS-PAGE and autoradiography. The appearance of these phosphoproteins was blocked by pretreatment of the membranes with H-7 or sangivamycin, two selective inhibitors of PKC. In addition, membranes purified from B cells treated with an inactive phorbol ester or stimulated with dibutyryl cAMP failed to exhibit a pattern of new phosphoproteins. These results are consistent with the involvement of PKC in the phosphorylation of the proteins. These phosphoproteins are also candidates for proteins whose functions are modified as a consequence of early signal delivery to resting B cells following membrane immunoglobulin occupancy. This system was utilized to identify the heavy chain of MHC class I molecules as one of the membrane proteins phosphorylated by PKC. The MHC class II molecules were not phosphorylated in membranes isolated from PMA-treated normal B cells or from PMA-treated B cells which had previously been exposed to IL-4. These results indicate that class I, but not class II, MHC molecules are phosphorylated by PKC. It is possible that such a modification of cell surface class I molecules may be involved during the process of signal transduction leading to B cell activation.

Animals

Organization and expression of a second chromosome follicle cell gene cluster in Drosophila.

Four genes expressed during the period of vitelline membrane formation are clustered within 8 kb of DNA in region 26A of the second chromosome. Temporal and quantitative difference in the profiles of accumulated RNA suggest that the genes are independently regulated although they are selectively expressed during the stages of vitelline membrane biosynthesis. In situ hybridization and S1 analyses of RNAs from fractionated eggchambers established that these genes are active only in the follicle cells. S1 mapping with in vitro synthesized RNA probes shows that three of the genes are tandemly oriented. All four appear to be intronless. In vitro translation products from hybrid-selected RNAs indicate that two of these genes code for major vitelline membrane proteins. Sequence analysis of these two genes support this conclusion. The cell- and stage-specific expression of the other two genes, encoding less abundant RNAs, suggests that they also play a role in early eggshell production.

Amino Acid Sequence

Bilateral simultaneous rupture of the quadriceps tendons. A report of four cases and review of the literature.

Four patients with bilateral simultaneous rupture of the quadriceps tendons were each older than 70 years of age. In the first patient the diagnosis was missed and one side was managed conservatively, while the other side was surgically treated. A combination of inadequate postoperative immobilization and the use of chromic catgut sutures probably caused failure of the repair, requiring revision surgery. The other three patients were treated by bilateral operative repair and had satisfactory results. Nonabsorbable suture material use and a minimum period of six weeks of postoperative immobilization are recommended.

Aged

Characterization and sequence of follicle cell genes selectively expressed during vitelline membrane formation in Drosophila.

To isolate genes involved in vitelline membrane production, an ovarian cDNA library was screened with eggchamber RNAs labeled in vivo. Two cDNA clones encoding RNAs that are selectively expressed in follicle cells during the period of vitelline membrane formation were isolated. Following isolation of homologous genomic clones from a Drosophila library, one gene was localized by in situ hybridization to chromosomal region 26A, and the other to 3C. Developmental Northern blots demonstrated that both genes produce 700-800 nucleotide transcripts that accumulate during the stages of vitelline membrane synthesis. In vitro translation products from hybrid selected RNAs and DNA sequence analysis both indicate that the 26A region gene encodes a major protein component of the vitelline membrane. The structural properties of the 3C region follicle cell gene seem more compatible with an intracellular function.

Amino Acid Sequence

The treatment of retroperitoneal fibromatosis with medroxyprogesterone acetate.

Wide excision is the recommended primary therapy for retroperitoneal fibromatosis. Radiation therapy and a variety of medications have been used to treat patients with recurrent tumors, but the response to these agents has not been uniform. The patient presented was successfully treated with medroxyprogesterone acetate for recurrent retroperitoneal fibromatosis that was refractory to multiple operative resections and radiation therapy.

Adult