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Biomedical subjects

D A Morrison

Publications and source records attributed to D A Morrison.

At least 127 records · Page 7Linked to original sources

Isolation and characterization of three new classes of transformation-deficient mutants of Streptococcus pneumoniae that are defective in DNA transport and genetic recombination.

Transformation-deficient mutants of Streptococcus pneumoniae were isolated after nitrosoguanidine mutagenesis. Seventeen mutants developed normal peaks of competence, as tested by their ability to degrade one strand of donor DNA, but they yielded transformants for chromosomal point markers at efficiencies from less than 0.001 to 0.04 that of the wild type. Some of the mutants were defective in DNA uptake and are described as entry defective (Ent-). Others took up DNA in normal quantities, but they failed to give stable transformants and are described as recombination defective (Rec-). In two of the Rec- mutants, normal levels of transformation by plasmid DNA occurred; in the others, it was reduced as much as chromosomal transformation. Conjugative transfers of a chromosomal omega (cat tet) element and of the plasmid pIP501 occurred at normal levels both to and from Rec- mutants. Transfer of chloramphenicol resistance by transformation with omega (cat tet) donor DNA, however, was blocked in Rec- mutants to about the same extent as was transformation for point markers.

Conjugation, Genetic↗

Fate of DNA in eclipse complex during genetic transformation in Streptococcus pneumoniae.

Uptake of DNA and genetic recombination proceeded normally in competent Streptococcus pneumoniae despite inhibition of DNA replication by 6-(p-hydroxyphenylazo)-uracil. Immediately after a brief uptake period, 68% of donor DNA label was in eclipse complex form, and 22% was in low-molecular-weight products; by the completion of integration at 10 min, 23% was integrated into the chromosome, and the rest was lost from the cell. Throughout the process, less than 1% was found as free single strands. The DNA in eclipse complex is therefore an intermediate in the integration process.

DNA Replication↗

Competence for genetic transformation in pneumococcus depends on synthesis of a small set of proteins.

In bacterial genetic transformation the uptake of DNA and its integration into the resident chromosome is dependent on a special cellular state, termed competence. In those species where appearance of competence has been studied, specific (but often poorly defined) growth conditions lead to a simultaneous development of competence in a substantial fraction of the cells in a culture. In Bacillus subtilis, and in Haemophilus species, competence appears in the stationary phase of growth or in certain other growth-limiting conditions. Streptococcus pneumoniae (pneumococcus) is perhaps unusual in that virtually all cells of a culture become competent, for a short period at a specific cell density during logarithmic growth, without perturbing the growth rate. The synchronous appearance of competence in pneumococcal cultures results from an autocatalytic effect of a small protein released by the cells that induces competence. The response to competence factor has been shown to require protein synthesis. We report here additional information on the nature of competence in pneumococcus: pulse-labelling studies show that for the brief period of competence protein synthesis is restricted to a few specific polypeptides.

Bacterial Proteins↗

Transformation in pneumococcus: nuclease resistance of deoxyribonucleic acid in the eclipse complex.

Donor deoxyribonucleic acid strands in the eclipse phase of genetic transformation of pnuemococcus (Streptococcus pneumoniae) are purified as a complex with a cf the deoxyribonucleic acid strand in this complex to digestion by nucleases was shown to be 50- to 1,000-fold less than that of uncomplexed single strands of deoxyribonucleic acid. Deoxyribonuclease I, micrococcal nuclease, Neurospora endonuclease, nuclease P1, and the major endogenous nuclease of cell-free extracts were studied. Sensitivity to nuclease attack was not uniform along the deoxyribonucleic acid strand; sequences of strongly protected bases were separated by more sensitive regions. The minimum size of protected fragments was about 70 bases. A complex of protein with the protected deoxyribonucleic acid segments was obtained after partial digestion. The sizes of these complexes, of the protected deoxyribonucleic acid segments, and of the protein subunit released by complete nuclease digestion, are all approximately identical, as determined by gel exclusion chromatography. Deoxyribonucleic acid strands of eclipse complex were also shown to be particularly well protected from attack by the major pneumococcal endonuclease in cell extracts.

Bacterial Proteins↗

Radiographic patterns of drug-induced lung diseases.

Drugs which induce lung disease are categorized according to the radiographic pattern of the disease. The five categories are: (a) diffuse interstitial (reticulo-nodular) findings, (b) diffuse air-space consolidation, (c) pleural effusion of fibrosis, (d) hilar or mediastinal widening, and (e) localized areas of consolidation. Information regarding onset, reversibility, fever, eosinophilia, and findings associated with each drug is presented. An extensive list of references is included.

Drug-Related Side Effects and Adverse Reactions↗

Biopsy of the main carina; staging lung cancer with the fiberoptic bronchoscope.

The efficacy of main carinal biopsy through the fiberoptic bronchoscope for evaluating resectability has not been determined. Forty-eight patients with carcinoma, but without gross neoplastic involvement of the main carina, underwent biopsy. Five (10 percent) had abnormal results on biopsy of the main carina. Three of the five patients were initially considered candidates for surgery. There were no complications from the procedure. Biopsy of the main carina through the fiberoptic bronchoscope is a valuable staging procedure in selected patients because of its simplicity and yield and the significance of abnormal findings.

Adenocarcinoma↗

Transformation in pneumococcus: protein content of eclipse complex.

A two-step purification of pneumococcal eclipse complex is described, which uses sucrose gradient sedimentation followed by agarose gel permeation chromatography. Purified complex contains, in addition to donor DNA single strands, macromolecular material that can be labeled with methionine or leucine during development of competence. This material co-chromatographed with eclipse complex DNA on hydroxylapatite, was dissociated from the DNA by sodium dodecyl sulfate, and was completely digested by Pronase. The sodium dodecyl sulfate-released material eluted as a single peak in sodium dodecyl sulfate chromatography. These properties were consistent with the noncovalent association with eclipse complex of a protein or class of proteins with a narrow range of polypeptide sizes. Evidence for the specific association of this protein with transforming DNA is eclipse was also obtained from parallel purification from 35S-labeled nontransformed cells; the amount of methionine label in the corresponding fractions in such cells was only 5% of that in transformed cells.

Bacterial Proteins↗

Vatican II and pluralism in pastoral care.

The documents of Vatican II imply that the ever increasing plurality of needs in the world and in Catholic health care institutions must be met by plurality in Christian response. Catholic hospitals should welcome onto their pastoral care teams people with diverse credentials and use them to promote the spiritual care of patients--Catholic and non-Catholic. In addition, this pluralistic ministry should extend itself beyond institutional walls toward the social needs of the community.

Catholicism↗

Transformation in pneumococcus: existence and properties of a complex involving donor deoxyribonucleate single strands in eclipse.

Donor deoxyribonucleic acid (DNA) single strands exist in a complex during the eclipse phase in pneumococcal transformation. This eclipse complex exhibited specific physical properties distinct from those of both pure DNA single strands and native DNA. These included a lower affinity for diethylaminoethyl-cellulose and hydroxylapatite than that of single-strand DNA, faster sedimentation than the DNA chains that it contains, and a buoyant density in Cs2SO4 lower than that of native DNA. The complex was dissociated by treatments with sodium dodecyl sulfate, NaOH, guanidine-hydrochloride, chloroform, and proteinase K but was insensitive to ribonuclease.

Centrifugation, Isopycnic↗

Structure of deoxyribonucleic acid on the cell surface during uptake by pneumococcus.

We exposed competent cells of Diplococcus pneumoniae to high-molecular-weight donor deoxyribonucleate (DNA) and examined the state of the DNA bound to them in forms sensitive to deoxyribonuclease I. The portion elutable with 5 M guanidine hydrochloride was shown to be native, of much lower molecular weight (4 x 10(6) to 5 x 10(6)) than the donor, and as active in further transformation as sheared DNA of the same size. The portion resistant to release by guanidine hydrochloride was also shown to be native and active in transformation. These results, along with previous ones, imply that the breaks produced outside the cell are not at genetically specific sites. Furthermore, it was found that entry past the cell barrier to deoxyribonuclease could occur at 0 C by a process sensitive to ethylenediaminetetraacetate.

Carbon Isotopes↗

Activity of deoxyribonucleic acid fragments of defined size in Bacillus subtilis transformation.

The transforming activity of Bacillus subtilis deoxyribonucleic acid (DNA) that had been sheared and purified with respect to size by sucrose gradient sedimentation is given as a function of the DNA molecular weight. It is shown (i) that fragments of median molecular weight 1.2 x 10(6) have finite activity (10(-4)), (ii) that the shape of the activity-versus-molecular weight function is qualitatively similar to that observed previously for Diplococcus pneumoniae, and (iii) that this shape precludes interpretation in terms of critical size models.

Bacillus subtilis↗

Transformation and deoxyribonucleic acid size: extent of degradation on entry varies with size of donor.

The fate of label introduced as donor deoxyribonucleic acid (DNA) into competent cells of Diplococcus pneumoniae was determined immediately after entry at 25 C, as a function of the size of the donor DNA. Part of the label is found to be acid soluble, part has been incorporated into chromosomal DNA, apparently through reincorporation of degraded donor DNA, and part is found in single strands of length smaller than that of the input donor DNA strands. The last fraction apparently constitutes the precursor for integration of intact donor genetic markers and is referred to as the intact fraction. For large donor DNA the intact fraction contains over 80% of the total intracellular label, but the median strand length has been reduced to 2.2 x 10(6) daltons. For small donor molecules (1 x 10(5) to 6 x 10(5) daltons per strand) the fraction intact increases with donor size from 10 to 50% of the total intracellular label, and the median strand length of this fraction is half that of the donor strands. By combining these results with earlier data on the size dependence of the yield of transformants per unit of total intracellular donor label, we have calculated the probability that a marker in the intact fraction will be integrated, as a function of the length of the donor strand after entry. This probability has a linear dependence on strand length for activities below 40% of maximum, and extrapolates to zero activity at 77,000 daltons per strand.

Bacillus subtilis↗

Early intermediate state of transforming deoxyribonucleic acid during uptake by Bacillus subtilis.

Examination of the effects of ethylenediaminetetraacetic acid (EDTA) on uptake of transforming deoxyribonucleic acid (DNA) by Bacillus subtilis allowed definition of a new intermediate state of transforming DNA during uptake. Markers in this state, termed "EDTA-resistant," are bound to the cell, are sensitive to inactivation by deoxyribonuclease, but may become deoxyribonuclease-resistant despite the addition of excess EDTA. Markers become EDTA-resistant quadratically with time after the addition of DNA to a competent culture, but linearly after exposure of competent cells to a brief pulse of DNA. An attractive model consistent with these findings is that EDTA blocks only the initiation of entry of a molecule of DNA.

Bacillus subtilis↗