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Biomedical subjects

J B Boyd

Publications and source records attributed to J B Boyd.

126 records · Page 7Linked to original sources
PubMed1976

Characterization of postreplication repair in mutagen-sensitive strains of Drosophila melanogaster.

Mutants of Drosophila melanogaster, with suspected repair deficiencies, were analyzed for their capacity to repair damage induced by X-rays and UV radiation. Analysis was performed on cell cultures derived from embryos of homozygous mutant shocks. Postreplication repair following UV radiation has been analyzed in mutant stocks derived from a total of ten complementation groups. Cultures were irradiated, pulse-labeled, and incubated in the dark prior to analysis by alkaline sucrose gradient centrifugation. Kinetics of the molecular weight increase in newly synthesized DNA were assayed after cells had been incubated in the presence or absence of caffeine. Two separate pathways of postreplication repair have been tentatively identified by mutants derived from four complementation groups. The proposed caffeine sensitive pathway (CAS) is defined by mutants which also disrupt meiosis. The second pathway (CIS) is caffeine insensitive and is not yet associated with meiotic functions. All mutants deficient in postreplication repair are also sensitive to nitrogen mustard. The mutants investigated display a normal capacity to repair single-strand breaks induced in DNA byX-rays, although two may possess a reduced capacity to repair damage caused by localized incorporation of high specific activity thymidine-3H. The data have been employed to construct a model for repair of UV-induced damage in Drosophila DNA. Implications of the model for DNA repair in mammals are discussed.

Animals↗
PubMed1976

The mei-9 alpha mutant of Drosophila melanogaster increases mutagen sensitivity and decreases excision repair.

The mei-9(a) mutant of Drosophila melanogaster , which reduces meiotic recombination in females (Baker and Carpenter 1972), is deficient in the excision of UV-induced pyrimidine dimers in both sexes. Assays were performed in primary cultures and established cell lines derived from embryos. An endonuclease preparation from M. luteus , which is specific for pyrimidine dimers, was employed to monitor UV-induced dimers in cellular DNA. The rate of disappearance of endonuclease-sensitive sites from DNA of control cells is 10-20 times faster than that from mei-9(a) cells. The mutant mei-218, which is also deficient in meiotic recombination, removes nuclease-sensitive sites at control rates. The mei-9(a) cells exhibit control levels of photorepair, postreplication repair and repair of single strand breaks. In mei-9 cells DNA synthesis and possibly postreplication repair are weakly sensitive to caffeine. Larvae which are hemizygous for either of the two mutants that define the mei-9 locus are hypersensitive to killing by the mutagens methyl methanesulfonate, nitrogen mustard and 2-acetylaminofluorene. Larvae hemizygous for the mei-218 mutant are insensitive to each of these reagents. These data demonstrate that the mei-9 locus is active in DNA repair of somatic cells. Thus functions involved in meiotic recombination are also active in DNA repair in this higher eukaryote. The results are consistent with the earlier suggestions (Baker and Carpenter 1972; Carpenter and Sandler 1974) that the mei-9 locus functions in the exchange events of meiosis. The mei-218 mutation behaves differently in genetic tests and our data suggest its function may be restricted to meiosis. These studies demonstrate that currently recognized modes of DNA repair can be efficiently detected in primary cell cultures derived from Drosophila embryos.

Animals↗
PubMed1975

The proteins of polytene chromosomes of Drosophila hydei.

It is reported that chromatin can be prepared from highly purified polytene nuclei from the salivary glands of third instar larvae of Drosophila hydei; such chromatin differs from that of diploid nuclei mainly by deficiencies in certain nonhistone chromosomal proteins. It is suggested that these proteins are important components of constitutive heterochromatin, which is severely underrepresented in polytene chromosomes. Chromosome morphology, including the pattern of induced puffs, is maintained throughout the mass isolation of glands and sucrose gradient purification of nuclei, as indicated by studies on temperature-shocked and control larvae. No significant alteration in the chromosomal proteins following puff induction by heat shock could be detected on analysis of the isolated protein fractions by disc gel electrophoresis. More sensitive techniques must be developed to study the apparent rearrangement or accumulation of protein at puff sites, and to elucidate the role of this protein in gene activation.

Animals↗
PubMed1974

Repair replication and photorepair of DNA in larvae of Drosophila melanogaster.

Repair replication of DNA has been studied in first instar larvae of Drosophila melanogaster with isopycnic centrifugation techniques. Larvae were fed BUdR, FUdR, streptomycin, penicillin, and Fungazone for two to four hours prior to exposure to UV, X-rays, MMS, or EMS. Feeding was continued for four hours in the presence of (3)HBUdR and DNA was isolated from whole larvae. Repair replication is stimulated by each of these agents. MMS is about 10 times as potent as EMS in stimulating repair synthesis. A dose of 200 ergs/mm(2) largely saturates the level of repair replication observed after UV irradiation. Repair replication rises between 0 and 80,000 R of X-rays before falling off. Semiconservative synthesis is seriously inhibited above a dose of 40,000 R of X-rays. Photorepair has been detected as a reduction in repair synthesis resulting from post-irradiation exposure to photoreactivating light. The same treatment has no detectable effect on X-ray-stimulated repair replication. Repair replication is insensitive to the presence of caffeine or hydroxyurea during the final incubation, although semiconservative synthesis is strongly inhibited by these agents. A mixture of BUdR and (3)HTdR can be used to replace (3)HBUdR in detecting repair replication.

Animals↗
PubMed1969

Structural and functional properties of polytene nuclei isolated from salivary glands of Drosophila hydei.

Salivary gland nuclei of Drosophila hydei, isolated by a modification of the procedure described by Boyd et al. (9), retain their normal morphology during the isolation and subsequent incubation procedure. RNA synthesis was studied in isolated nuclei by biochemical and cytological techniques. In radioautographs 70% of the nuclei displayed a distribution of labeled RNA over the nuclear constituents similar to the distribution obtained after in vivo incorporation of radioactive precursor. Chromosome puffs and the nucleoli were specifically labeled. The remaining 30% of the nuclei showed a weak to very weak incorporation of radioactive precursor. In these nuclei most of the radioautographic grains were concentrated over the nucleolus, and a few grains were randomly distributed over the chromosomes. Actinomycin D and the absence of ATP, GTP, and CTP in the medium inhibited incorporation of radioactive precursor. The radioactive product was sensitive to combined pronase and RNase digestion. Addition of E. coli RNA polymerase to the incubation medium enhanced the specific labeling over the puffed regions. The sedimentation behavior of the RNA synthesized in isolated nuclei was different from that of RNA synthesized during a 20 min pulse of radioactive precursor administered to whole glands in vivo and in vitro. Neither the steroid ecdysterone nor a temperature treatment was effective in inducing new puffs in isolated nuclei.

Adenosine Triphosphate↗
PubMed1968

Mass preparation of nuclei from the larval salivary glands of Drosophila hydei.

A method has been developed for isolating gram quantities of salivary glands from late third instar larvae of Drosophila hydei. The isolated glands have a normal appearance and incorporate RNA and DNA precursors normally. Nuclei can be isolated from these glands in 90% yield with the use of detergents. These nuclei contain morphologically normal giant polytene chromosomes.

Animals↗
PubMedDate not supplied

Tumors of the skull base: outcome and survival analysis of 77 cases.

We reviewed our experience with combined approaches to lesions that transcend the bones of the skull base. Seventy-seven skull base procedures were performed on 73 patients during a 10-year period from 1982 to 1992. There were 34 patients (44%) with region lesions (anterior), seven patients (9%) with region II lesions (anterior-lateral), 25 patients (32%) with region III lesions (lateral-posterior), and 11 patients (14%) with lesions that invaded more than one anatomic site. The histopathology in this series was quite variable, with 22 patients (29%) having squamous cell carcinoma and eight patients (10%) having basal cell carcinoma. Forty-one patients had surgery by an anterior approach and 38 patients had lateral approaches, with 18 undergoing an infratemporal approach and 29 undergoing temporal bone resections. Overall, 44% of the patients had a postoperative complication. Survival of this heterogeneous group of patients is 79% at 2 years and 71% at 4 years, with those patients with region II disease having a statistically significant poorer prognosis with no survivors at 4 years.

Adolescent↗
PubMedDate not supplied

Third-chromosome mutagen-sensitive mutants of Drosophila melanogaster.

A total of 34 third chromosomes of Drosophila melanogaster that render homozygous larvae hypersensitive to killing by chemical mutagens have been isolated. Genetic analyses have placed responsible mutations in more than eleven complementation groups. Mutants in three complementation groups are strongly sensitive to methyl methanesulfonate, those in one are sensitive to nitrogen mustard, and mutants in six groups are hypersensitive to both mutagens. Eight of the ten loci mapped fall within 15% of the genetic map that encompasses the centromere of chromosome 3. Mutants from four of the complementation groups are associated with moderate to strong meiotic effects in females. Preliminary biochemical analyses have implicated seven of these loci in DNA metabolism.

Animals↗