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Biomedical subjects

D Lloyd

Publications and source records attributed to D Lloyd.

At least 19 recordsLinked to original sources
PubMed1995

Temperature-dependent changes in plasma-membrane lipid order and the phagocytotic activity of the amoeba Acanthamoeba castellanii are closely correlated.

The relationship between temperature-dependent changes in phagocytotic activity of Acanthamoeba castellanii and the fatty acid composition and physical properties of plasma membrane-enriched fractions were determined in cultures acclimated to 30 degrees C and 15 degrees C. Chilling (from 30 degrees C to 15 degrees C) had a very pronounced short-term inhibitory effect on phagocytosis only in stationary-phase cultures, which displayed a low degree of fatty acid unsaturation. A subsequent increase in phagocytosis by these cells was correlated with a low-temperature-induced increase in fatty acid unsaturation (shown previously [Jones, Lloyd and Harwood (1993) Biochem. J. 296, 183-188] to be due to n-6 desaturase induction). Plasma membrane-enriched fractions from 15 degrees C-acclimated cells also showed a marked increase in the relative proportion of polyunsaturated fatty acids. Steady-state fluorescence depolarization studies, using the membrane probe diphenylhexatriene, revealed increases in plasma membrane order with decreasing assay temperature. Over the upper assay-temperature range (25-40 degrees C), fluorescence anisotropy values were higher in membranes from 30 degrees C-acclimated cells; a 3.3 degrees C relative displacement of plots indicated that temperature-induced changes in membrane lipid composition compensated for approx. 22% of the ordering effect of low temperature. Changes in the temperature-dependence of fluorescence anisotropy, possibly corresponding to lateral phase separations or alterations in other bulk physical properties of membranes, occurred between 20 and 25 degrees C in membranes from 30 degrees C-acclimated cells and between 15 and 20 degrees C in membranes from 15 degrees C-acclimated cells. Fluorescence anisotropy plots were superimposed at assay temperatures between 5 and 15 degrees C. Short-term phagocytotic rates in whole cells decreased with assay temperature. Arrhenius discontinuities in rates of phagocytosis occurred at approx. 25.0 degrees C and 17.5 degrees C in 30 degrees C- and 15 degrees C-acclimated cells respectively, and in each case were thus within the temperature ranges of slope-change in the corresponding fluorescence anisotropy plots. The results show a direct correlation between plasma membrane fatty acid unsaturation, membrane physical properties and phagocytotic activity in A. castellanii. Therefore, a specific integrated physiological process has been correlated with fatty acid desaturase induction for the first time.

Acanthamoeba↗
PubMed1995

The effects of co-cultivation with the acetogen Acetitomaculum ruminis on the fermentative metabolism of the rumen fungi Neocallimastix patriciarum and Neocallimastix sp. strain L2.

The effects of co-cultivation with the hydrogen-utilizing acetogenic bacterium Acetitomaculum ruminis on the fermentative activities of the rumen fungi Neocallimastix patriciarum or Neocallimastix sp. L2 were investigated. In both co-cultures acetate production increased, making it the predominant fermentation product, as the accumulation of lactate, formate, ethanol, H2 and (in the case of Neocallimastix sp. L2) succinate all decreased. The effects of co-cultivation with Methanobrevibacter smithii were more pronounced. Decreased activities of lactate dehydrogenase, alcohol dehydrogenase and (in the case of Neocallimastix sp. L2) fumarate reductase accompanied the shift in fermentation product formation. The rate of glucose utilization and the fungal biomass yield were also increased in the co-culture.

Acetates↗
PubMed1995

The aniline blue fluorochrome specifically stains the septum of both live and fixed Schizosaccharomyces pombe cells.

A novel method is described which uses aniline blue for the specific fluorescent staining of the septa of dividing cells of the fission yeast, Schizosaccharomyces pombe. It gives the same results with live and fixed cells. In fixed or, more generally, dead cells there is no staining of the cytoplasm: this renders aniline blue superior to other dyes previously used to stain the septum of S. pombe. This feature allows quantitative analysis of the septum index for fixed samples and, therefore, makes aniline blue the stain of choice for cell cycle kinetic studies.

Aniline Compounds↗
PubMed1995

Cell cytotoxicity of sodium nitrite, sodium nitroprusside and Roussin's black salt against Trichomonas vaginalis.

We have investigated the action of sodium nitrite and other nitrosyl complexes, such as sodium nitroprusside and Roussin's black salt, on the growth of metronidazole-sensitive and resistant strains of Trichomonas vaginalis and their hydrogenosomal enzymes. All three chemicals inhibited the growth of T. vaginalis: sodium nitrite at 8 mM, sodium nitroprusside at 1.2 mM and Roussin's black salt at 0.2 mM. Metronidazole-sensitive (KT9) and resistant (CDC85) isolates showed similar cytotoxicity against these molecules. Specific activities of pyruvate:ferredoxin oxidoreductase and hydrogenase and oxygen uptake rates were decreased in the T. vaginalis isolate treated with sodium nitrite and sodium nitroprusside. However, Roussin's black salt increased the specific activity of pyruvate:ferredoxin oxidoreductase or hydrogenase in CDC85 or KT9 cells and increased the oxygen uptake rate in the KT9 isolate.

Animals↗
PubMed1995

A temperature-compensated ultradian clock ticks in Schizosaccharomyces pombe.

An ultradian oscillation is described for Schizosaccharomyces pombe which meets the criteria for a cellular clock, i.e. timekeeping device. The rhythm can be induced by transfer from circadian conditions (stationary phase or very slow growth) to ultradian conditions (rapid growth). It can also be synchronized by ultradian temperature cycles of 6 degrees C difference. Released to constant temperature, the rhythm persists for 20 h without damping. The period of the free-running rhythm is temperature-compensated and in no experiment did period length fall outside the narrow range between 40 and 44 min. The parameter observed is the septum index, i.e. the percentage of cells occupying the last stage of the cell cycle in wild-type cells before final division. The results suggest control of the cell division processes by the ultradian clock.

Activity Cycles↗
PubMed1995

The ability of membrane potential dyes and calcafluor white to distinguish between viable and non-viable bacteria.

Various dyes were assessed for their ability to discriminate between viable and non-viable bacteria. Two methods of killing were employed: by heat treatment or by gramicidin treatment. Staining was carried out in two ways; by staining directly in the medium or by washing cells prior to staining in buffer. Carbocyanine and rhodamine 123 dyes only exhibited small changes in fluorescence between viable and non-viable populations of bacteria. Both oxonol dye (bis 1,3-dibutylbarbituric acid trimethine oxonol) and calcafluor white proved much more useful.

Bacterial Physiological Phenomena↗
PubMed1995

Scottish neonatal intensive care units; a study of staff and parental attitudes.

The study takes a three stage approach to review Scottish unit policies, staff views and parents experiences of visiting and involvement with their low birthweight infant (1,750 gm or less) in a sample of Scottish neonatal intensive care units (NICUs). The study indicated that while most units had 'liberal' views regarding parental and family visiting some staff views were not in line with their unit policy and retained reservations about family visiting. Many parents reported visiting daily despite considerable 'social' and real costs. Unit differences emerged regarding the degree of parental involvement as measured by tasks carried out.

Adult↗
PubMed1994

Effects of oxygen, pH and nitrate concentration on denitrification by Pseudomonas species.

The production of nitrogen-containing gases by denitrification in three organisms was examined using membrane inlet mass spectrometry. The effects of O2 (during both growth and maintenance) and of pH, nitrate concentration and carbon source were tested in non-proliferating cell suspensions. Two strains of Pseudomonas aeruginosa were capable of co-respiration of NO3- and O2 and, under controlled O2 supply, gave oscillatory denitrification. Variations in culture and assay conditions affected both the rate of denitrification and the ratio of end products (N2O:N2). Higher rates were seen following anaerobic growth. Optimum values of pH and nitrate concentration for denitrification are given. Generally, the optimum pH was 7.0-7.5, approximately that of the growth medium. Optimum nitrate concentration was generally 20 mM.

Acetates↗
PubMed1994

Determination of the viability of Trichomonas vaginalis using flow cytometry.

In clinical laboratories, viability of Trichomonas vaginalis is determined by using light microscopy (differential count of motile to nonmotile organisms). Alternative methods are proposed that utilise flow cytometry. Under an epifluorescence microscope, live organisms fluorescence intensely green with fluorescein diacetate (FDA), whereas dead cells fluoresce orange with propidium iodide (PI). Flow cytometric histograms of green versus red fluorescence reveal distinct populations for live and dead cells. The anionic oxonal probe DiBAC4(3) is a membrane potential sensitive dye that distributes between the inside of the cell and the medium. Live organisms are less fluorescent than dead organisms when stained with the oxonol probe. Valinomycin, dicyclohexylcarbodiimide, and vanadate all give significant changes in the fluorescence intensities of cultures stained with the oxonol probe compared with control cultures, indicating that this probe is detecting changes in plasma membrane potential. Both FDA/PI and oxonol staining protocols allow good discrimination between populations and permit counts that are more statistically significant than those obtained by light microscopy. These methods remove the subjectiveness of microscopic counts and would increase the accuracy of susceptibility assays.

Animals↗
PubMed1994

Polypeptides of hydrogenosome-enriched fractions from rumen ciliate protozoa and trichomonads: immunological studies.

The evolution of hydrogenosomes, energy-generating organelles of rumen ciliate protozoa and the flagellate trichomonads has been the subject of much speculation. Polypeptides of the hydrogenosome-enriched fractions from the rumen ciliates, Dasytricha ruminantium, Isostricha spp., Polyplastron multivesiculatum and Eudiplodinium maggii were separated by SDS-PAGE and compared to analogous polypeptide preparations from Tritrichomonas foetus. Immunoblotting with antisera specific to the hydrogenosomes of T. foetus identified common immunoreactive polypeptides present at estimated molecular masses of 28, 35, 38, 44, 48, 58, 100 and 120 kDa. That at 120 kDa corresponds to a single subunit of the purified pyruvate:ferredoxin oxidoreductase from the hydrogenosome of Trichomonas vaginalis.

Animals↗