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Biomedical subjects

D C White

Publications and source records attributed to D C White.

At least 145 records · Page 8Linked to original sources

The role of magnesium adenosine triphosphate in the contractile kinetics of insect fibrillar flight muscle.

The changes in tension produced by small step or sinusoidal changes of length have been measured for chemically skinned flight muscle fibres of the giant tropical water bug Lethocerus at MgATP concentrations in the range 0.1-10 mM. In the presence of calcium ion concentrations of about 20 microM, the rates of the rapid mechanical processes observed were found to increase with increasing MgATP, exhibiting saturation with an apparent half-saturation constant lying between 0.1 and 1 mM MgATP, depending upon the conditions used. Under the same conditions, an increase in MgATP concentration was found to lead to a slight decrease in the isometric tension generated by the preparations. The results are discussed with reference to some current crossbridge models of muscle contraction.

Actomyosin↗

Biomass measurement of methane forming bacteria in environmental samples.

Methane-forming bacteria contain unusual phytanylglycerol ether phospholipids which can be extracted from the bacteria in sediments and assayed quantitatively by high performance liquid chromatography (HPLC). In this procedure the lipids were extracted, the phospholipids recovered, hydrolyzed, purified by thin layer chromatography, derivatized and assayed by HPLC. Ether lipids were recovered quantitatively from Methanobacterium thermoautotrophicum and sediments at levels as low as 8 x 10(-14) moles. In freshwater and marine sediments the flux of methane to the atmosphere and the methane levels in the pore water reflects the recovery of the phytanyl glycerol ether lipid 'signature'. The proportion of the ether phospholipid to the total recoverable phospholipid was highest in anaerobic digester sewage sludge and deeper subsurface freshwater sediment horizons.

Biomass↗

Servo-controlled closed-circuit anaesthesia. A method for the automatic control of anaesthesia produced by a volatile agent in oxygen.

A method of closed-circuit anaesthesia has been developed in which the end-tidal concentration of the volatile anaesthetic agent used is controlled automatically using a closed-loop servo system. End-tidal anaesthetic concentrations, measured by the Engstrom EMMA, were maintained in the closed circuit by direct liquid injection. The system was tested in the laboratory and in clinical use (12 subjects). Accurate estimation of anaesthetic uptake was readily obtained and results for the uptake of halothane agreed closely with those of previous workers. The major sources of error in the method were the result of zero offset in the Engstrom EMMA which in turn was caused by humidity and the intrinsic characteristic of the simple proportional controller used. These errors were easily correctable, and end-tidal halothane concentration could be controlled to within 0.1%. Mean halothane vapour uptake at a constant end-tidal concentration of 0.8% was 114 ml min-1 at 1 min, 36 ml min-1 at 5 min, 29 ml min-1 at 10 min and between 22 and 18 ml min-1 at 20-35 min.

Adult↗

Uptake of isoflurane during clinical anaesthesia. Servo-control of liquid anaesthetic injection into a closed-circuit breathing system.

Twenty-six patients were anaesthetized with isoflurane (Forane) in oxygen using a completely closed-circuit breathing system to which liquid isoflurane was injected. The rate of injection was controlled to maintain the end-tidal concentration at a pre-set value (1.5% = 1.3 MAC). The rate of injection of anaesthetic was recorded and from the data the uptake of anaesthetic by the patient was measured. It was found to average 44.5 ml of vapour at 2 min, 23.7 ml at 10 min and 19.4 ml at 30 min. Attempts were made to correlate the data with anthropometric measurements.

Adolescent↗

The elasticity of relaxed insect fibrillar flight muscle.

The mechanical properties of glycerol-extracted fibres from the dorsal longitudinal muscle of Lethocerus have been determined by sinusoidal and transient analysis in the time range 1 ms-1000 s, and from rest length to 10% strain for fibres in relaxing and rigor solutions. The fibres behave reversibly up to strains of about 5%, but reach an elastic limit in the range 5-9% strain, depending upon the rate of strain. Electron micrographs of fibres at different degrees of stretch, and after partial extraction of the contractile proteins, suggest that a connexion between the end of the A filament and the Z line, named a C filament, is responsible for the high stiffness of the relaxed muscle. Estimates are made of the compliance of the A, I and C filaments. The mechanical response of the relaxed muscle, over the entire frequency range studied, is assignable to the C filaments. An analysis of the stiffness of the fibres at different tensions in activating and relaxing solutions, and in fibres relaxed by orthovanadate, shows that the C filaments still exert their mechanical effect in the active muscle. That is, the response of the active muscle consists of the contribution from the cross-bridges plus that of the C filaments, acting mechanically in parallel. This situation is incompatible with earlier explanations of the fully activated mechanical dynamics of fibrillar muscle. Alternative explanations at the cross-bridge level are described in the paper that follows this one.

Animals↗

Role of cross-bridge distortion in the small-signal mechanical dynamics of insect and rabbit striated muscle.

The mechanism of the active tension response of insect fibrillar muscle to step changes and small oscillations of length was re-investigated, following White's demonstration (1983) that the high relaxed stiffness evidently persists during activation and cannot be neglected as had previously been assumed. White's result makes earlier explanations of the small-signal response untenable; the experimental and theoretical studies described here lead to a new class of explanations at the cross-bridge level. The response of an activated muscle to a fast stretch consists of a synchronous tension increase that is followed first by a rapid decay of tension and then by a delayed rise ('stretch activation'). It was shown in glycerinated fibre preparations from the water bug and the bumblebee that subtraction of the relaxed tension response from the active response results in a prominent undershoot of the tension level preceding the step, before the delayed rise of tension. The responses of the same fibres to sinusoidal oscillations, in the frequency range 1-150 Hz, showed an equivalent behaviour, with the active locus circling the relaxed locus in a Nyquist plot, as described by Machin & Pringle (1960). Stiffness was determined during the tension response to a large step (of 1%) by recording the immediate change of tension to a small test step (0.2%), applied at various times after the conditioning step. In the majority of preparations stiffness remained constant or increased during the undershoot of tension. Step and sinusoidal responses with the above features cannot be explained at all by an active component resembling a simple exponential delay. We show, however, that such features are predicted if certain small-signal effects of cross-bridge distortion (previously and erroneously assumed negligible in insect flight muscle for the small-signal case) are incorporated in models of the cross-bridge cycle. Two alternative hypotheses for the effects of distortion are examined: (i) distortion-induced detachments and (ii) distortion-modulated transitions among multiple attached states (Huxley & Simmons, 1971). For the first we also show that the results do not differ qualitatively whether one assumes strain, interfilament displacement or 'bridge recruitment' as the physical correlate of stretch activation. Both of the above explanations account, at least qualitatively, for the observed rapid decay and undershoot of tension following a step increase of length, and for the circling of the active Nyquist-plot loci about the passive locus. The explanation based on distortion-induced detachments, however, appears to be inconsistent with the stiffness measurements.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Spatial distribution of biochemical parameters indicating biomass and community composition of microbial assemblies in estuarine mud flat sediments.

The spatial distribution of communities was examined in estuarine mud flat sediments by the biochemical analysis of the lipids and lipid components extracted from the sediments. Total phospholipid was used as a measure of total biomass, and fatty acids were used as indicators of community composition. Comparisons were made among 2- by 2-m (location) and 0.2- by 0.2-m (cluster) sampling plots by using a nested analysis of variance to design an optimal sampling strategy to define the microbial content of a large, relatively homogenous area. At two of the three stations, a 2- by 2-m plot was representative of the station, but 0.2- by 0.2-m areas were in no case representative of the station. The biomass measured by the extractable phospholipid and the total lipid palmitic acid showed excellent correlation with the fatty acid "signatures" characteristic of bacteria, but showed a lower correlation with the long-chain polyenoic fatty acids characteristic of the microfauna.

Journal Article↗

Relationship Between Physiological Status and Formation of Extracellular Polysaccharide Glycocalyx in Pseudomonas atlantica.

Marine pseudomonads, such as Pseudomonas atlantica, are readily isolated from sediments. These organisms form extracellular polysaccharide polymers (glycocalyx). The factors affecting the composition and amount of glycocalyx in batch culture of these organisms were examined. The formation of glycocalyx was stimulated by the inclusion of galactose as the carbon source and by increased surface area resulting from addition of sand to the medium. The composition of the glycocalyx changed during the growth cycle, with a marked increase in the proportions and absolute amounts of uronic acids as the rate of synthesis increased. In estuarine sediments, the glycocalyx contained a carbon content at least as great as in the microbes themselves. The greatest accumulation of these polymers occurred late in the stationary phase when the physiological status of the cells, as measured by the adenylate energy charge, showed maximal stress. Maximal formation of glycocalyx possibly could be used as an estimate of the nutritional status of these microbes.

Journal Article↗

Polymeric Beta-Hydroxyalkanoates from Environmental Samples and Bacillus megaterium.

The procaryotic endogenous storage polymer known as poly-beta-hydroxybutyrate is actually a mixed polymer of short-chain beta-hydroxy fatty acids. A method for the quantitative recovery of this mixed polymer, called poly-beta-hydroxyalkanoate (PHA), with analysis by capillary gas-liquid chromatography, showed the presence of at least 11 short-chain beta-hydroxy acids in polymers extracted from marine sediments. Polymers extracted from Bacillus megaterium monocultures were also a complex mixture of beta-hydroxy acids with chain lengths between four and eight carbons. Lyophilized sediments were extracted in a modified Soxhlet extractor, and the polymer was purified with ethanol and diethyl ether washes. The purified polymer was treated with ethanol-chloroform-hydrochloric acid (8.5:2.5:1) for 4 h at 100 degrees C, a treatment which resulted in the formation of the ethyl esters of the constituent beta-hydroxy acids. Subsequent assay of the products by gas-liquid chromatography indicated excellent reproducibility and sensitivity (detection limit, 100 fmol). Disturbing sediments mechanically or adding natural chelators increased all major PHA components relative to the bacterial biomass. Gardening of sedimentary microbes by Clymenella sp., an annelid worm, induced decreases in PHA, with changes in the relative proportion of component beta-hydroxy acids. The concentration of PHA relative to the bacterial biomass can reflect the recent metabolic status of the microbiota.

Journal Article↗

Effects of oil and gas well-drilling fluids on the biomass and community structure of microbiota that colonize sands in running seawater.

Well-drilling fluid and a number of the known components (barite, clay, Aldacide, Surflo, and Dowicide, were tested for effects on the biomass and community structure of the microbiota that colonize marine sands exposed for eight weeks to running ambient seawater. Shading the microbiota from light depressed the microflora without a significant effect on the biomass, while well-drilling fluids layered on the surface or mixed with the sand significantly increased a component of the bacteria and the microfauna as reflected in changes in the fatty acid composition. There were some shading effects from the surface layering of well-drilling fluids as reflected in the fatty acids from the microflora when compared to the sands mixed with well-drilling fluids. Barite had essentially no effect on the biomass or community structure while clays increased nearly all of the biomass indicators for the bacteria as well as the microfauna; the clay overlay mirrors the effect of the drilling fluids. Aldacide shifted the bacterial composition, depressing the proportions of microbes containing the cyclopropane fatty acids and the anaerobic pathways of desaturation. Concentrations of 1 and 15 microgram/L increased the bacterial biomass as reflected in the total lipid (16:0) and extractable lipid phosphate coupled with a decrease in the total microeukaryotes. Surflo increased the biomass and shifted the bacterial community structure at concentrations between 4 and 800 microgram/L. The lowest level also stimulated the microfauna. Dowicide at 100 microgram/L increased the bacteria forming cis-vaccenic acid and the microfauna similar to low concentrations of Surflo.

Fatty Acids↗

Estimations of uronic acids as quantitative measures of extracellular and cell wall polysaccharide polymers from environmental samples.

The extracellular polysaccharide polymers can bind microbes to surfaces and can cause physical modification of the microenvironment. Since uronic acids appear to be the components of these extracellular films that are most concentrated in a location outside the cell membrane, a quantitative assay for uronic acids was developed. Polymers containing uronic acids are resistant to quantitative hydrolysis, and the uronic acids, once released, form lactones irreproducibly and are difficult to separate from the neutral sugars. These problems were obviated by the methylation of the uronic acids and their subsequent reduction with sodium borodeuteride to the corresponding alcohol while they were in the polymer and could not form lactones. This caused the polymers to lose the ability to adhere to their substrates, so they could be quantitatively recovered. The hydrolysis of the dideuterated sugars was reproducible and could be performed under conditions that were mild enough that other cellular and extracellular polymers were not affected. The resulting neutral sugars were readily derivatized and then were separated and assayed by glass capillary gas-liquid chromatography. The dideuterated portion of each pentose, hexose, or heptose, identified by combined capillary gas-liquid chromatography and mass spectrometry, accurately provided the proportion of each uronic acid in each carbohydrate of the polymer. Examples of the applications of this methodology include the composition of extracellular polymers in marine bacteria, invertebrate feeding tubes and fecal structures, and the microfouling films formed on titanium and aluminum surfaces exposed to seawater.

Journal Article↗

Preservation of estuarine sediments for lipid analysis of biomass and community structure of microbiota.

Various methods were tested for preserving estuarine sediments in the field before biochemical analysis of the microbiota. Total microbial biomass was determined as lipid phosphate (LP), and the fatty acids of the microbial lipids were used as indicators of community structure. Control samples were sieved to remove macroinvertebrates and plant materials and were extracted immediately in the field. Other samples were preserved both before and after sieving and stored for 5 days before analysis. Freezing resulted in a 50% decline in LP and significant decreases in many fatty acids. Refrigeration resulted in a 19% decrease in LP but no change in the fatty acids. Samples preserved with Formalin before sieving exhibited no significant change in LP but substantial increases in many fatty acids, which were probably derived from the macroinvertebrates. Sieved samples preserved with Formalin showed a 17 to 18% decline in LP but no change in the fatty acids. Ideally, samples should be sieved and extracted immediately in the field. However, short-term refrigeration and longer-term preservation of sieved samples with Formalin may be acceptable compromises.

Journal Article↗

Sensitive assay, based on hydroxy fatty acids from lipopolysaccharide lipid A, for Gram-negative bacteria in sediments.

Biochemical measures have provided insight into the biomass and community structure of sedimentary microbiota without the requirement of selection by growth or quantitative removal from the sediment grains. This study used the assay of the hydroxy fatty acids released from the lipid A of the lipopolysaccharide in sediments to provide an estimate of the gram-negative bacteria. The method was sensitive to picomolar amounts of hydroxy fatty acids. The recovery of lipopolysaccharide hydroxy fatty acids from organisms added to sediments was quantitative. The lipids were extracted from the sediments with single-phase chloroform-methanol extraction. The lipid-extraction residue was hydrolyzed in 1 N HCl, and the hydroxy fatty acids of the lipopolysaccharide were recovered in chloroform for analysis by gas-liquid chromatography. This method proved to be about fivefold more sensitive than the classical phenol-water or trichloroacetic acid methods when applied to marine sediments. By examination of the patterns of hydroxy fatty acids, it was also possible to help define the community structure of the sedimentary gram-negative bacteria.

Bacillus subtilis↗

Effect of silicate grain shape, structure, and location on the biomass and community structure of colonizing marine microbiota.

Microbiota colonizing silica grains of the same size and water pore space, but with a different microtopography, showed differences in biomass and community structure after 8 weeks of exposure to running seawater. The absence of surface cracks and crevices resulted in a marked diminution of the total microbial biomass measured as lipid phosphate and total extractable palmitic acid. With increasing smoothness of the sand grain surface, examination of the community structure showed a marked decrease in procaryotes and algal microeucaryotes, with a relative increase in microeucaryotic grazers. A comparison of the colonizing sediment incubated in running seawater or at 32 m on the sea floor with a sediment core showed a decreased bacterial biomass with a different community structure and a decreased total microeucaryotic population of both grazers and algae. The quantitative differences in microbial biomass and community structure between the microcosms and the actual benthic population in the core were determined.

Journal Article↗

Effect of manual brush cleaning on biomass and community structure of microfouling film formed on aluminum and titanium surfaces exposed to rapidly flowing seawater.

Metals exposed to rapidly flowing seawater are fouled by microbes that increase heat transfer resistance. In this study, results of biochemical test methods quantitatively relating the biomass and community structure of the microfouling film on aluminum and titanium to heat transfer resistance across the metal surface during three cycles of free fouling and manual brushing showed that cleaning accelerates the rate of fouling measured as the loss of heat transfer efficiency and as microfouling film biomass. The results also showed that the rate of fouling, measured as an increase in heat transfer resistance, is faster on titanium than on aluminum but that the titanium surface is more readily cleaned. In three cycles of free fouling and cleaning with a stiff-bristle nylon brush, the free-fouling communities re-forming on aluminum became enriched in bacteria containing short-branched fatty acids as the cycling progressed. The free-fouling community on titanium revealed an increasingly diverse morphology under scanning electron microscopy that was enriched in a portion of the microeucaryotes. Brushing removed most of the biomass, but left a residual community that was relatively enriched in a portion of the bacterial assembly containing cyclopropane fatty acids on aluminum and in a more diverse community on the titanium surface. The residual communities left after cleaning of titanium revealed an increase in bacteria with short-branched fatty acids and in microeucaryotes as cleaning continued. No significant changes occurred in the residual microbial community structure left on aluminum with cleaning; it was, again, less diverse than that remaining on titanium. The residual communities secreted a twofold-larger amount of extracellular polymer, measured as the ratio of total organic carbon to lipid phosphate, than did the free-fouling community on both surfaces.

Journal Article↗

Effect of light on biomass and community structure of estuarine detrital microbiota.

Comparison of estuarine detrital microbiota grown with and without light in the absence of macroscopic grazing showed shifts in the community structure that enabled correlation between various biochemical measures. Analysis of these biochemical measures showed that growth in light induces the smallest increases in procaryotic attributes such as muramic acid; wall glucosamine; lipid phosphate; total extractable adenosine nucleotides; short-branched, cyclopropane, and cis-vaccenic fatty acids; lipid glucose and mannose; the incorporation of acetate into lipid; and the formation of deoxyribonucleic acid from thymidine. Measures of the microfauna such as lipid inositol and the gamma-linolenic series of polyenoic fatty acids also increased minimally in the light-grown microbiota. Measures of sulfo-lipid synthesis, lipid glycerol, total extractable palmitate, 18-carbon polyenoic fatty acids, and total polyenoic fatty acids longer than 20 carbons increased 10- to 15-fold in algae and fungi. Chlorophyll a, lipid galactose, and the 16- and 20- carbon polyenoic fatty acids characteristic of diatoms increased maximally in the light. This increase of diatom measure correlated with the sheets of diatoms detected by scanning electron microscopy.

Journal Article↗

Segmental pancreas transplantation from living related and cadaver donors: a clinical experience.

Between July 25, 1978, and January 25, 1981, 20 segmental pancreas transplants from 14 cadaver (cad) and six related (rel) donors were performed. All but two recipients had previously received renal allografts for treatment of end-sage diabetic nephropathy. The grafts were placed intraperitoneally-13 with open ducts, 4 with prolamine-injected ducts, and 3 with silicone rubber-injected ducts. At this writing six patients have functioning grafts (3 silicone, 3 open duct) at less than 1 (cad), 2 (cad), 3 (rel), 15 (cad), 20 (rel), and 31 (cad) months. Five open-duct grafts failed for technical reasons. Two prolamine-injected grafts lost function between 1 and 3 months; biopsy specimens showed severe fibrosis. The others lost function between 2 and 4 months. Three patients died 1 to 3 months after transplantation. Metabolic test results were normal or nearly normal in most recipients with functioning grafts. Splenectomy was required in one related donor; there were no other complications of donation. Technical problems and rejection are impediments to application of pancreas transplantation, but a sustained correction of the metabolic defect has been achieved in a few patients, and the effect on secondary complications are studied.

Adolescent↗