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Biomedical subjects

H Swift

Publications and source records attributed to H Swift.

At least 37 records · Page 2Linked to original sources

Quantitative cytochemistry of the diaminobenzidine cytochrome oxidase reaction product in mitochondria of cardiac muscle and pancreas.

The rate constant (k) of the cytochrome oxidase reaction under optimal conditions for cytochemical staining (i.e., 15 min fixation, incubation for 180 min for heart, 120 min for pancreas) can be used as a measure of the enzyme concentration within mitochondria. The rate constant derived from microdensitometric measurements of the mass thickness of the 3,3'-diaminobenzidine (DAB) cytochrome oxidase reaction in cristae times correlated data derived from morphometry on the surface density of cristae (SVcristae/Vmit micron-1) and the volume density of mitochondria per cell (Vmit/Vcell) has been used to determine the respiratory index (RI) of these tissues according to the following equation: RI = k(SVcristae/Vcell). Using this formula, the RI of cardiac muscle tissue was computed to be 33 times the RI of pancreas under the conditions of our experiments. The greater cristae surface density and the large mitochondrial volume density in cardiac muscle and high k value accounted for the higher RI of cardiac muscle.

3,3'-Diaminobenzidine↗

Marital interaction and adjustment in patients with essential hypertension.

Twenty-five men and twenty-six women with essential hypertension, and their spouses, were compared with normotensive control couples on a questionnaire measure of marital adjustment. The marriages of the hypertensive men showed significantly raised levels of marital dissatisfaction in the spouses, and an abnormal degree of reciprocal dissatisfaction between spouses. The marriages of the female patients showed an abnormal lack of reciprocity between spouses' scores. These findings support anecdotal reports of a relationship between marital disharmony and communication problems and elevated blood pressure. It is suggested that clinicians could routinely assess marital adjustment in patients with essential hypertension, and that in some cases conjoint marital therapy might be an adjunct or alternative to anti-hypertensive medications.

Adult↗

Alteration of protein synthesis and induction of specific protein phosphorylation by hyperthermia.

Confluent cultures of the mouse cell line clone 1D were subjected to 1-hr hyperthermic treatments. Temperatures were increased from the control level of 37 degrees to values ranging from 38 to 45 degrees. Protein synthesis patterns were determined in fluorograms of sodium dodecyl sulfate-polyacrylamide gels labeled with [3H]leucine. Although incorporation into most proteins was either repressed or decreased by the treatment, several proteins showed an increased label of were apparently induced de novo. Among the induced proteins was a prominent band, probably a doublet, with an estimated molecular weight of 70,000 to 69,000. Crude cell lysates made from 37 degrees, 41 degrees, and 45 degrees-treated cells were tested for kinase activity at 30 degrees by a 10-min incubation with adenosine [gamma-32P]triphosphate. Several specific proteins exhibited increased phosphorylation, while phosphorylation of other proteins decreased. The most significant increase in phosphorylation was shown by a protein with molecular weight of about 37,000. We suggest that heat treatment induces or activates one or more specific phosphokinase(s) with the ability to phosphorylate proteins with approximate molecular weights of 37,000, 36,000, 23,000, and 16,000.

Animals↗

Morphology of a novel cyanobacterium and characterization of light-harvesting complexes from it: Implications for phycobiliprotein evolution.

The morphology of the marine cyanobacterium DC-2 and two light-harvesting complexes from it have been characterized. DC-2 has an outer cell wall sheath not previously observed, the purified phycoerythrin shows many unusual properties that distinguish it from all phycoerythrins characterized to date, and isolated phycobilisomes have a single absorption band at 640 nm in the phycocyanin-allophycocyanin region of the spectrum. On the basis of these observations we suggest that DC-2, rather than being a member of the Synechococcus group, should be placed in its own taxonomic group. In addition, the particular properties of the isolated phycoerythrin suggest that it may be representative of an early stage in the evolution of the phycoerythrins. These observations are of special interest in light of the contribution DC-2 and related cyanobacteria may make to global primary productivity.

Journal Article↗

Pigment mutants of Chlamydomonas with deformed plastids and increased levels of chloroplast nucleic acids.

The chloroplast ultrastructure of two Chlamydomonas reinhardtii pigment variant mutants, U3N and U3A, is strikingly different from that of the wild type. The mutant chloroplast has greatly lowered levels of chlorophyll a and b, and lacks the usual ordered thylakoid membrane structure. The amount of chloroplast ribosomes is increased, but the pyrenoid and surrounding starch grains appear to be unaltered. Our biochemical analyses have shown that, while the properties of chloroplast DNA, ribosomal RNA, and ribosomes in these mutants appeared to be normal, their relative amounts per cell increased markedly when compared to the wild type. In U3N these increases were approximately 60% for chloroplast DNA and 80% for chloroplast ribosomes. However, the ratio of chloroplast rDNA genes to total chloroplast DNA remained unchanged as shown by DNA-rRNA hybrdization. We propose that (1) The enhanced level of chloroplast ribosomes in these mutants is a direct consequence of the elevated amount of chloroplast DNA. Both of these increases may, in turn, arise from defective mechanism for their control. (2) These mutants grow successfully in the absence of functional photosynthesis, provided an external carbon source is available to them, but functional plastid DNA, ribosomes and protein synthesis may still be a requirement for normal starch metabolism.

Chlamydomonas↗

Genomic organization in the flesh fly Sarcophaga bullata.

The genome of the flesh fly Sarcophaga bullata has been characterized both cytologically and biochemically. S. bullata has a haploid DNA level of 0.61 picograms which is five times larger than the haploid genome size of Drosophila melanogaster. Reassociation kinetics of Sarcophaga DNA shows that its sequence organization is very similar to that of D. melanogaster in having a very large proportion of single copy DNA (81%) and only small amounts of highly and moderately repetitive DNA (9% and 6%, respectively). cRNAs from all three sequence classes were prepared and their cytological distributions on biploid and polytene cells determined by in situ hybridization. The cytological distribution of the highly repetitive probe was found to be restricted to the centromeric heterochromatin of two of the five autosomes and this sequence class was also found to be markedly underreplicated in polytene foot-pad cells. No highly repetitive DNA was localized on either of the sex chromosomes, but only on the two large centromeric regions of chromosomes C and E. Moderately repetitive DNA was found uniformly distributed on all of the autosomes in both testis and polytene foot-pad squashes. As in the case of the highly repetitive sequence probe, no moderately repetitive DNA was detected on either the X or Y chromosomes. Moderately repetitive DNA in Sarcophaga was also shown to have the "Drosophila type" pattern of sequence interspersion with a moderately repetitive element of congruent to 5,000 nucleotides adjacent to a unique element of greater than 10,000 nucleotides. The Sarcophaga genome is the largest for which this type of interspersion has so far been demonstrated.

Animals↗

Characterization of extrachromosomal DNA in the flesh fly Sarcophaga bullata.

The polytene pupal foot pad cells of the flesh fly Sarcophaga bullata contain numerous extrachromosomal DNA containing granules. We have determined both the origin and the nature of the DNA sequences present in these granules. Studies done with quinacrine staining of seven day old pupal foot-pad polytene nuclei showed that the granules fluoresced very brightly while the chromosomal bands to which the granules were attached did not. The only other highly fluorescent regions of the polytene karyotype were the centromeric heterochromatin of chromosomes C and E and several bands associated with the nucleolus of Chromsome A. When polytene nuclei were hybridized in situ with cRNA made from highly repetitive DNA, many of the granules positively labeled. Most of the label on these slides was concentrated on the centromeric heterochromatin of chromosomes C and E. Quinacrine staining of the foot-pad cells at very early stages of pupal development showed that when granules were present, they were always closely associated with the same two centromeric regions, those of chromosomes C and E. Since the highly repetitive DNA located in these centromeric regions is underreplicated, we conclude that the granules result from an extrusion process which takes place early during the polytenization of these cells. The chromosomal integrity of the centromeric heterochromatin of chromosomes C and E is apparently disrupted and repetitive sequences are dissociated from the chromosomes as DNA granules which then secondarily become associated with chromosomal bands throughout the nucleus.

Animals↗

Isolation and characterization of mitochondrial DNA from Chlamydomonas reinhardtii.

Mitochondrial DNA (mtDNA) has been isolated from a mitochondrial pellet of Chlamydomonas reinhardtii. The mtDNA has a buoyant density of 1.706 g/ml in CsCl, a melting temperature of 87.9 degrees in standard saline citrate, and a nucleoside composition of 47.5% deoxyguanidine plus deoxycytidine with no odd nucleosides. Thermal denaturation and renaturation studies have shown that (i) mtDNA contains no extensive intramolecular heterogeneity nor significant base bias between the complementary polynucleotide chains and (ii) mtDNA renatures as a single homogeneous class with a kinetic complexity of 9.78 X 10(6) daltons. Although rare (less than or equal to 1%), both open and supercoiled circular mtDNA molecules have been observed in the electron microscope. Contour lengths of linear and open and closed circular molecules are all within the range of 4.0-5.4 micron with a mean of 4.67 +/- 0.30 micron. This size is similar to that of animal mtDNA but approximately 1/8 that of the higher plant mtDNAs. The magnitude of mtDNA reiteration in C. reinhardtii is estimated to be of the same order as that of chloroplast DNA.

Centrifugation, Isopycnic↗

Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma. I. Ultrastructural analysis of organelles.

Electron microscope studies have been made on the fine structure of the colorless biflagellate, Polytoma obtusum, with main emphasis on the structural organization of the mitochondria and the leucoplast. Both organelles have been demonstrated to contain DNA aggregates as well as ribosomal particles within their matrix material. Reconstructions from serial showed that (a) the mitochondria were highly convoluted and irregular in shape and size, and (b) the leucoplast was a single cup-shaped entity, with large starch grains, localized at the posterior end, and multiple sites of DNA aggregates. The starch-containing compartments appeared to be interconnected by narrow tubular or sheetlike bridges. Cytoplasmic invaginations into the plastid, often containing mitochondria, were of frequent occurrence, and membranes of mitochondria and the leucoplast appeared to be closely apposed. Membranes elements, both sheetlike and vesicular, were also present in the matrix. The Polytoma leucoplast was, in certain respects, morphologically similar to the plastids of various photosynthetic mutants of Chlamydomonas, most of which show Menedelian segregation. It is suggested that Polytoma arose from a Chlamydomonas-like ancestor, possibly through combined mutational processess of both chloroplast and nuclear genomes. Since Polytoma leucoplasts contain both DNA and ribosomal particles, it is probable that these organelles still possess semiautonomy and limited ability for protein synthesis.

Cell Membrane↗

Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma. II. General characterization of organelle nucleic acids.

Polytoma obtusum has a main band DNA (alpha) with a buoyant density in CsC1 of rho = 1.711 g/ml and a light DNA satellite (beta) with rho = 1.682 g/ml. beta-DNA was substantially enriched in a fraction containing small leucoplast fragments and some mitochondria, which was obtained in a pellet sedimenting between 3,000 g and 5,000 g. A crude mitochondrial pellet was also obtained by sedimenting at 12,000 g to recover particulates remaining in the supernate after 10 min at 5,000 g. This fraction contained a third DNA component (gamma) with rho = 1.714 g/ml. We have concluded that the leucoplasts of P. obtusum contain the beta-DNA (1.6882) and the mitochondria possess the gamma-component (1.714). Two distinct classess of ribosomes were isolated and separated by sucrose density gradients, a major 79S species and a minor species at 75S. The major species possessed the 25S and 18S ribosomal RNA (rRNA), characteristic of cytoplasmic ribosomes, and these particles co-sedimented in sucrose gradients with the 79S cytoplasmic ribosomes of Chlamydomonas reinhardtii. The minor species was present in about 2% of the total ribosomal population but showed an eight-to-ninefold enrichment in the leucoplast pellet, suggesting that it was of organelle origin. These 73S particles had RNA components migrating very closely with the 18S and 25S species of the 79S ribosomes, but the base composition of the rRNA from these two classes of ribosomes was significantly different; the rRNA from the 79S ribosomes had a G+C mole ratio of 50.0%, while the rRNA from the 73S class had a ratio of 47.5%. By comparison, chloroplast ribosomes of C. reinhardtii were found to sediment at 70S and contain rRNA molecules of 23S and 16S, with a G + C content of 51.0%. These findings support the concept that the Polytoma leucoplast possesses characteristic genetic and protein-forming systems.

Chlamydomonas↗

Characterization of cytoplasmic and nuclear genomes in the colorless alga Polytoma. III. Ribosomal RNA cistrons of the nucleus and leucoplast.

The colorless alga Polytoma obtusum has been found to possess leucoplasts, and two kinds of ribosomes with sedimentation values of 73S and 79S. The ribosomal RNA (rRNA) of the 73S but not the 79S ribosomes was shown to hybridize with the leucoplast DNA (rho - 1.682 g/ml). Nuclear DNA of Polytoma (rho = 1.711) showed specific hybridization with rRNA from the 79S ribosomes. Saturation hybridization indicated that only one copy of the rRNA cistrons was present per leucoplast genome, with an average buoyant density of rho = 1.700. On the other hand, about 750 copies of the cytoplasmic rRNA cistrons were present per nuclear genome with a density of rho = 1.709. Heterologous hybridization studies with Chlamydomonas reinhardtii rRNAs showed an estimated 80% homology between the two cytoplasmic rRNAs, but only a 50% homology between chloroplast and leucoplast rRNAs of the two species. We conclude that the leucoplasts of Polytoma derive from chloroplasts of a Chlamydomonas-like ancestor, but that the leucoplast rRNA cistrons have diverged in evolution more extensively than the cistrons for cytoplasmic rRNA.

Biological Evolution↗