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Biomedical subjects

J D Harvey

Publications and source records attributed to J D Harvey.

At least 19 recordsLinked to original sources

AIDS after Montreal.

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Acquired Immunodeficiency Syndrome

Electron donor properties of the antitumour drug amsacrine as studied by fluorescence quenching of DNA-bound ethidium.

The effect of the antitumour acridine derivative amsacrine [4'-(9-acridinylamino)methanesulphon-m-anisidide] on the fluorescence lifetime of DNA-bound ethidium has been investigated using a synchronously pumped cavity dumped dye laser producing picosecond pulses for sample excitation and a time-correlated single photon counting detection system. As the proportion of DNA-bound amsacrine on the synthetic DNA polymer poly[deoxyadenylic-thymidylic acid] is increased, the fluorescence decay curve of ethidium can be accurately resolved into two exponential components. The short lifetime component, whose proportion increases with increasing proportions of DNA-bound amsacrine, has a lifetime of between 3 and 4 ns, significantly longer than that of ethidium in aqueous solution (1.63 ns). The magnitude of the long lifetime component decreases from 25.4 to 14 ns with increasing proportions of bound amsacrine. It is concluded that a new fluorescence state of ethidium (lifetime 3-4 ns) is present, probably resulting from reversible electron transfer between ethidium and amsacrine. The ability of various 9-anilinoacridine derivatives to quench the fluorescence of DNA-bound ethidium appears to be related to the electron donor properties of the substituents on the anilino ring, as well as to experimental antitumour activity. The electron donor properties of DNA-bound amsacrine may therefore be relevant to its antitumour action.

Acridines

Aerobic and anaerobic swimming speeds of spermatozoa investigated by twin beam laser velocimetry.

The motility of bovine and ovine spermatozoa has been studied under aerobic and anaerobic conditions, using a dual beam laser velocimeter. Cells swimming under aerobic conditions were found to be characterized by a translational swimming speed and a rotation rate that were approximately double those of cells swimming in an anaerobic environment. Both types of spermatozoa have been found to exhibit a sudden coordinated transition between fast and slow swimming states when the available oxygen is exhausted. This transition from aerobic to anaerobic swimming states has also been shown to be reversible. Studies of the duration of aerobic motility using the same apparatus have shown that the cells have a constant motile efficiency over the temperature range 32 degrees-42 degrees C.

Aerobiosis

Reovirus RNA transcriptase: evidence for a conformational change during activation of the core particle.

Reovirus cores contain an RNA transcriptase capable of synthesizing messenger RNA. When cores are suspended in 1 X SSC at 37 degrees they are quiescent and synthesize no product, but in the presence of the components of an RNA transcriptase reaction mixture they actively synthesize mRNA. Photochemical crosslinking has been used to investigate the arrangement of RNA and protein in both "quiescent" and "active" cores. Irradiation induces the formation of a noncovalent RNA:protein complex in "quiescent" but not in "active" cores. This difference is attributed to a conformational change in the reovirus core which results from the transition between the "quiescent" and "active" states of the particle.

Enzyme Activation

Twin-beam laser velocimeter for the investigation of spermatozoon motility.

Previous laser light-scattering studies of spermatozoon motility have been hampered by the large, asymmetric shape of spermatozoa, which causes difficulties in the interpretation of intensity fluctuations in the light scattered from a single laser beam. This paper describes an experimental arrangement for measuring the distribution of transit times for swimming spermatozoa using two slightly separated, focused laser beams. The theory of operation of the instrument is developed to enable the analysis of the experimentally obtained cross-correlation functions. The effects of the pronounced spermatozoon asymmetry and associated intensity modulation in the scattered light are also investigated and shown to be negligible for the twin beam experimental arrangement, provided that the swimming speed distribution has a coefficient of variation (sigma/upsilon greater than 0.1. Results obtained using this apparatus are presented for the velocity distribution of spermatozoa from a variety of bulls.

Animals

Light-scattering studies of bull spermatozoa. II. Interaction and concentration effects.

The complete autocorrelation function of the intensity fluctuations of laser light scattered from motile bull spermatozoa is shown to depend upon several factors not previously considered. Samples of bull spermatozoa generally contain a substantial proportion of dead cells, which give rise to slowly decaying components of the autocorrelation function. Whereas previous work has concentrated on the form of the fast decaying autocorrelation component, we are concerned here with the relative amplitude and shape of the slow autocorrelation component and the general form of the composite function. In principle, the relative amplitudes of the fast and slow components of the autocorrelation function can be used as an assay of the proportion of swimming cells. We show that this amplitude ratio depends upon cell concentration, scattering cell geometry, and scattering angle. A simple model is developed to explain these results on the basis of the asymmetry of light scattered from these cells, motile/immotile cell interactions, wall-swimming effects, and geotactic reorientation of dead cells.

Animals

The relationship between the size of mitochondria and the intensity of light that they scatter in different energetic states.

The intensity of light scattered at 90 degrees to the incident beam and the effective hydrodynamic radii of mitochondria incubated under a variety of conditions have been measured. Addition of high concentrations of uncouplers to respiring mitochondria resulted in a decrease in scatter which was not due to swelling. Addition of valinomycin to mitochondria depleted to substrate in K+-free medium produced an increase in scatter that was not due to shrinking. It is concluded that changes in the intensity of scattered light are not reliable indices of changes of volume of mitochondria, and the changes in conformation with changes in metabolic state dominate changes in light scatter. A molecular mechanism for the effect of metabolic state upon the scattered intensity is suggested.

Animals

Laser light-scattering studies of bull spermatozoa. I. Orientational effects.

Calculations based on the known dimensions of bull spermatozoa show that the scattered light intensity is strongly dependent upon the relative orientation of the particle to the incident beam. The magnitude of this effect of apparently much greater than for other systems where motility has been investigated by dynamic light scattering. The calculations show that the scattering source can be approximated by a small spinning mirror, and consequently the greatest light intensity at the detector results from cells swimming in a direction perpendicular to the scattering vector. The calculations are in substantial agreement with photographic observations, as well as direct measurements of the scattered intensity. Previous treatments of dynamic light scattering from swimming bull spermatozoa based on point scattering models are shown to be incorrect.

Animals

Subunit structure of the reovirus spike.

Cross-linking reovirus spike protein with the bifunctional reagent dimethyl suberimidate revealed that each spike was composed of a pentameric aggregate of polypeptide lambda 2.

Cross-Linking Reagents

Generation time statistics of Escherichia coli B measured by synchronous culture techniques.

Synchronous cultures of Escherichia coli B were produced under various environmental conditions. Analysis of the cell number data permitted the characterization of the generation time distribution for these organisms and the estimation of the mother-daughter generation time correlation coefficients. For all growth conditions, the distribution of generation times was found to be symmetrical with a coefficienoefficient was significantly negative at doubling times between 40 and 64 min. However, the results for a culture growing in succinate medium at 37 degrees C, which had a significantly greater generation time, yielded a correlation coefficient close to zero. Within the range of temperatures studied (26 to 37 degrees C), no significant effect on the correlation coefficient was observed.

Bacteriological Techniques

The molecular size and shape of liver glycogen.

The molecular-weight distribution of liver glycogen has been established from the analysis of sedimentation rates of fractions separated on sucrose density gradients and from the direct measurement of the diffusion coefficients of these fractions by laser-intensity-fluctuation spectroscopy. Hydrodynamic studies indicated that all fractions of glycogen of mol.wt.exceeding 25x10(6) had about 1.1 g of water per g of polysaccharide associated with them. The hydration and hydrodynamic behaviour of all fractions of mol.wt. exceeding 25x10(6) was similar, whereas smaller fractions behaved anomalously, indicating a substantially different overall structure.

Centrifugation, Density Gradient

Hydrodynamic properties of 2-mercaptoethanol-modified glycogen.

Treatment of glycogen with 2-mercaptoethanol and iodoacetamide gives rise to a modified glycogen which resembles the original glycogen in its hydrodynamic behaviour but has a pronounced tendency to aggregate. The modified glycogen can be distinguished easily, by its diffusion coefficient, from glycogen degraded by more traditional methods of extraction. The 'fundamental' glycogen particle appears to be composed of two or three glycogen beta-particles linked by a single protein chain.

Animals