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At least 145 records · Page 8Linked to original sources

Processing of secondary ion microscope images: an example of application to the thyroid.

Ion microscopy is a microanalytical method by which one can obtain distribution images of any chemical element with isotope discrimination even at very low local concentrations, in successive slices of the specimen. These images are obtained at the price of progressive erosion of the specimen, so that the analysis may not be replayed and it is necessary to record the maximum amount of information during specimen erosion. We present an improvement of this method using a highly sensitive camera connected to a video analog-digital converter. The images are acquired and digitized on line and may be processed by an image computer. We illustrate the technique described with an application of ion microscopy that is made possible by digital recording and processing of images. This application concerns the precise comparison of iodine isotopes and phosphorus distributions in sections of the thyroid gland of rats which were submitted to an iodine-deficient diet followed by an injection of 129I.

Animals↗

Molecular basis for the transformation defects in mutants of Haemophilus influenzae.

To determine the molecular basis of transformation defects in Haemophilus influenzae, the fate of genetically marked, (32)P-labeled, heavy deoxyribonucleic acid (DNA) was examined in three mutant strains (rec(1) (-), rec(2) (-), and KB6) and in wild type having (3)H-labeled DNA and a second genetic marker. Transforming cells upon lysis with digitonin followed by low-speed centrifugation are separable into the supernatant fraction, containing mainly the unintegrated donor DNA, and the pellet, containing most of the resident DNA along with integrated donor DNA. Electron micrographs of digitonin-treated cells also indicate that the resident DNA is trapped inside a cellular structure but that cytoplasmic elements such as ribosomes are extensively released. DNA synthesis in digitonin-treated cells is immediately blocked, as is any further integration of donor DNA into the resident genome. Isopycnic and sedimentation analysis of supernatant fluids and pellets revealed that in strains rec(2) (-) and KB6 there is little or no association between donor and resident DNA, and thus there is negligible transfer of donor DNA genetic information. In these strains, the donor DNA is not broken into pieces of lower molecular weight as it is in strain rec(1) (-) and in the wild type, both of which show association between donor and recipient DNA. In strain rec(1) (-), although some donor DNA atoms become covalently linked to resident DNA, the incorporated material does not have the donor DNA transforming activity.

Bacteriolysis↗

Molecular events accompanying the fixation of genetic information in Haemophilus heterospecific transformation.

Heterospecific transformation between Haemophilus influenzae and H. parainfluenzae was investigated by isopycnic analysis of deoxyribonucleic acid (DNA) extracts of (3)H-labeled transforming cells that had been exposed to (32)P-labeled, heavy transforming DNA. The density distribution of genetic markers from the resident DNA and from the donor DNA was determined by transformation assay of fractions from CsCl gradients, both species being used as recipients. About 50% of the (32)P atoms in H. parainfluenzae donor DNA taken up by H. influenzae cells were transferred to resident DNA, and only a small amount of the label was lost under conditions of little cell growth. There was less transfer in the reciprocal cross, and almost half of the donor label was lost. In both crosses, the transferred donor material transformed for the donor marker considerably more efficiently when assayed on the donor species than on the recipient species, indicating that at least some of the associated (32)P atoms are contained in relatively long stretches of donor DNA. When the transformed cultures were incubated under growth conditions, the donor marker associated with recipient DNA transformed the donor species with progressively decreasing efficiency. The data indicate that the low heterospecific transformation between H. influenzae and H. parainfluenzae may be due partly to events occurring before association of donor and resident DNA but results mostly from events that occur after the association of the two DNA preparations.

Bacteriolysis↗

Characterization of excision repair in Neurospora crassa.

The excision of pyrimidine dimers from the deoxyribonucleic acid (DNA) of Neurospora crassa was examined. Postirradiation incubation in the presence of several chemicals known to inhibit various repair systems indicated that caffeine reduced the rate of excision twofold, but did not inhibit excision completely as did proflavine and quinacrine. Examination of the time course of excision showed that repair occurs at a relatively rapid rate: approximately 60 dimers excised per min after 500 ergs/mm(2). Further evidence for rapid excision was obtained by sedimentation analysis of DNA; the maximal number of breaks introduced during repair was three, suggesting that breaks are repaired almost as fast as they are made and that only a few dimers are repaired at a time. Repair synthesis was measured by prelabeling the DNA with (15)N and D(2)O, and then subjecting the DNA to equilibrium density gradient centrifugation after postirradiation incubation with (32)P. Accumulation of single-strand breaks with increasing dose of ultraviolet radiation suggested that the limiting step was subsequent to the incision and excision steps of repair. Equilibrium CsCl centrifugation demonstrated that the limiting step in excision was repair synthesis.

Acridines↗

The relationship between plasma urea levels and some muscle trimethylamine levels in Xenopus laevis: a 31P and 14N nuclear magnetic resonance study.

Urea is known to be disruptive to proteins, yet high levels occur in a variety of tissues. It has been suggested that trimethylamines counteract the effects of urea on protein. We have, therefore, directly investigated whether elevating tissue urea levels produces an increase in trimethylamine concentrations by using 31P and 14N nuclear magnetic resonance (NMR) spectroscopy to detect two trimethylamines (glycine betaine and glycerylphosphorylcholine) in intact, living gastrocnemius muscle from Xenopus laevis. Xenopus laevis naturally elevates its tissue urea levels under dry conditions. This ability was used to obtain plasma urea levels ranging from 1 to 110 mmol l-1. This procedure did not alter pH or levels of ATP or phosphocreatine in the muscles, but did raise trimethylamine levels. We find that there is a significant relationship between plasma urea concentrations and the trimethylamine levels studied. This relationship was, however, limited to the lower range of urea levels. We propose that other trimethylamines or mechanisms come into play at urea concentrations above 20 mmol l-1.

Animals↗