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Biomedical subjects

C A Miles

Publications and source records attributed to C A Miles.

31 records · Page 2Linked to original sources

Attenuation of ultrasound in suspensions of bovine muscle myofibrils and myosin.

The attenuation of 1.5-7 MHz ultrasound was measured over the pH range 3-7 in 100 mM KCl suspensions of bovine M. semitendinosus myofibrils, precipitated myosin and the residue of myofibrils after partial extraction of myosin. In all fractions attenuation showed a similar dependence on pH over the range 3-7, with a broad, substantial maximum in the region of pH 4.5-pH 5.5 and similar mass attenuation coefficients (per g protein). At pH 7 and 7 MHz these were 3.49 +/- 0.20 cm2 g-1 in the myofibrils, 3.26 +/- 0.31 cm2 g-1 in the myofibrilar residue and 2.83 +/- 0.68 cm2 g-1 in the precipitated myosin. Measurements at 5.3 MHz of precipitated myosin over a wider pH range revealed an attenuation titration curve similar to that previously observed in homogenates of muscle and muscle myofibrils, with substantial peaks at about pH 5 and 11.5, and a shoulder perhaps indicating a small underlying peak at about pH 8-9. Myosin dissolved in 800 mM KCl gave attenuation levels that were typically 50% lower than precipitated myosin e.g. at pH 7 and 7 MHz: 2.83 +/- 0.68 cm2 g-1 in the precipitated form, 1.29 +/- 0.10 cm2 g-1 in solution. These results indicated that: (a) attenuation by myosin filaments contributed substantially to the total attenuation in suspensions of myofibrils and (b) the peak in the myofibrilar attenuation is caused, or substantially contributed to, by a process taking place in the myosin component.

Animals↗

Experimental estimation of the viscous component of ultrasound attenuation in suspensions of bovine skeletal muscle myofibrils.

It has been suggested that viscous losses, caused by the motion of myofibrils relative to their environmental fluid, could be the major cause of ultrasound attenuation in muscle. This Paper presents theoretical and experimental estimates of the viscous component of attenuation in suspensions of myofibrils. Experimental estimates were made by monitoring the effects of varying the viscosity and density of the suspending fluid and ranged from 0.55 to 0.72 cm2 g-1 protein at 7 MHz, when expressed as mass attenuation coefficients. This represented 16-22% of the total attenuation measured in suspensions. Corresponding theoretical calculations were lower: 0.44 and 0.21 cm2 g-1 protein, respectively, for suspensions with the myofibrils aligned along and across the direction of sound propagation. It was concluded that most of the attenuation was caused by other absorption processes.

Animals↗

Attenuation of ultrasound in homogenates of bovine skeletal muscle and other tissues.

The attenuation of ultrasound in homogenates of bovine skeletal muscle and suspensions of myofibrils was measured over the frequency range 1.5-7 MHz, and found to be proportional to protein concentration in both. In the homogenates it varied with frequency and temperature in a similar way to the attenuation in post rigor muscle tissue; myofibrils showed a higher frequency dependence. The attenuation in homogenates of bovine muscle, liver and kidney and in suspensions of myofibrils was measured over the pH range 3.5-13, and each showed a peak at about pH 11.5. This was thought to be due to a proton transfer process between NH3+ groups on the tissue proteins and OH- ions in the suspending fluid. A substantial peak at about pH 5 in the muscle and myofibril suspensions was not observed in homogenates of liver and kidney and was thought to be due to components of muscle that are absent from the other tissues. Myofibrils suspended in percoll solution of density 1.05 g cm-3, chosen to match approximately the density of the myofibrils, showed a slightly lower attenuation over the pH range 5-7, but a pH dependence similar to that of the myofibrils suspended in saline. The difference in the attenuations may be interpreted as the viscous component of the attenuation due to relative motion between the myofibril and its surrounding saline. The peak at pH 5 did not, however, appear to be due to the viscous loss mechanism peaking due to maximum shrinkage (and therefore maximum density) of the myofibril near this pH.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The relationship between the base composition of bacterial DNA and its intracellular melting temperature as determined by differential scanning calorimetry.

The correlation between the melting temperature of intracellular DNA, determined by differential scanning calorimetry (DSC) of whole bacteria, and its guanine + cytosine (G + C) content, was examined for 58 species of bacteria. Samples of vegetative cells were heated in a Perkin-Elmer DSC-2C at 10 degrees C min-1 from 5 to 130 degrees C, cooled to 5 degrees C and then re-heated as before. Literature values for the mole fraction of G + C, XGC, were linearly related to the temperature, Tmax, at which the reversible peak, pr, observed on the second heating run was at a maximum, via the equation XGC = (Tmax -73.8)/41.0. This equation accounted for 91.9% of the variance in XGC with 95% confidence limits of +/- 7.3%, approximately 1.6 times the corresponding uncertainty (+/- 4.5%) quoted by De Ley (Journal of Bacteriology 101, 738-754, 1970) for estimates based on the spectroscopically determined melting temperature of purified DNA. Random errors of measurement of Tmax did not greatly limit the precision of the prediction and it was concluded that factors additional to base composition affected the temperature of DNA melting within the bacterial cell. Displacement of Tmax values from the fitted line was particularly noticeable in Campylobacter, Corynebacterium and Bacterionema species and part of the residual variation appeared to be species specific, possibly caused by differences in intracellular solute concentration.

Base Composition↗

Attenuation of ultrasound in post rigor bovine skeletal muscle.

A pulse transmission method for measuring the attenuation of 1-7 MHz ultrasound in bovine skeletal muscle is described. Measurements of the attenuation coefficient at -20, 0, 20 and 40 degrees C conformed to the relation alpha = Afn, where A and n are temperature-dependent coefficients and f is the frequency. alpha/f varied slowly with frequency, and at 4 MHz and 20 degrees C mean values were 1.3 dB cm-1 MHz-1 along the fibres and 0.55 dB cm-1 MHz-1 across the fibres. These data are lower than most previous measurements of skeletal muscle, but comparable with recent measurements of canine heart muscle.

Animals↗

Ultrasound speed and attenuation in homogenates of bovine skeletal muscle.

The attenuation and speed of ultrasound were measured in homogenates of post-rigor bovine skeletal muscle, and found to increase in proportion to the concentration of muscle. Extrapolation of the data to tissue concentrations yielded an attenuation of 7.5 dB cm-1 at pH 5.7, 20 degrees C and 7.3 MHz. This was close to that measured in the minced tissue, 8.3 dB cm-1, and between values previously recorded across and along the fibres of intact muscle. Corresponding measurements for the speed of ultrasound in homogenates, extrapolated to the native tissue concentration, were: 1555 +/- 9 m s-1 at 0 degree C, 1592 +/- 10 m s-1 at 20 degrees C and 1616 +/- 9 m s-1 at 37 degrees C. These were not significantly different from measurements of minced muscle at the same temperatures. Measurements of the attenuation of 7.3 MHz ultrasound in suspensions of myofibrils indicated that attenuation by the myofibrils caused at least 64% of the attenuation in muscle homogenates at pH 5.7. Re-analysis of the viscous loss arising from relative movement of the myofibrils in their surrounding fluid, indicated that this mechanism could account for no more than 15% of the attenuation in muscle homogenates. Attenuation due to scattering was calculated to be at least two orders of magnitude smaller than that observed in either homogenates or suspensions of myofibrils. It was concluded that the contribution of scattering to the attenuation was small, and that the attenuation was caused by processes involving an absorption of energy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Differential scanning calorimetry of bacteria.

Thermograms obtained by differential scanning calorimetry of a range of bacteria of different heat resistances were compared. Equations were derived to calculate the rate at which the numbers of viable organisms in a calorimeter decline as the temperature is raised at a constant rate. Vegetative bacteria scanned at 10 degrees C min-1 showed multi-peaked thermograms with four major peaks (denoted m, n, p and q) occurring in the regions 68-73, 77-84, 89-99 and 105-110 degrees C respectively. Exceptions were that peak m (the largest peak) occurred at 79-82 degrees C in Bacillus stearothermophilus and an additional peak, r, was detected in Escherichia coli at 119 degrees C. At temperatures below the main peak m there were major differences in thermograms between species. There was a direct relationship between the onset of thermal denaturation and the thermoresistance of different organisms. Heat-sensitive organisms displayed thermogram features which were absent in the more heat-resistant types. When samples were cooled to 5 degrees C and re-heated, a small endothermic peak, pr, was observed at the same temperature as p. Peaks p and pr were identified as the melting endotherms of DNA. In all vegetative organisms examined, maximum death rates, computed from published D and z values, occurred at temperatures above the onset of thermal denaturation, i.e. cell death and irreversible denaturation of cell components occurred within the same temperature range.

Bacillus cereus↗

Transit of a meal through the stomach, small intestine, and colon in normal subjects and its role in the pathogenesis of diarrhea.

A method for measuring the transit time of a meal, containing sausages, mashed potato, baked beans, and a pineapple custard dessert, through the gastrointestinal tract was evaluated in 14 healthy volunteers. Gastric emptying was determined by incorporating a radioactive marker in the meal and counting over the surface of the stomach using a crystal scintillation detector. Small intestinal transit time was determined by measuring breath hydrogen excretion and by estimating the radioactivity over the cecum. Finally, whole gut transit time was measured by incorporating radiopaque plastic markers or carmine red in the meal and estimating the appearance of these markers in the stool. Our results showed that measurements of small intestinal transit time were reproducible and in the majority of subjects the increase in hydrogen excretion occurred at the same time as the increase in radioactive counts over the surface of the cecum. The passage of the first marker in the stool coincided with the appearance of carmine red. There were no significant correlations between small intestinal transit time and whole gut transit time or the half time for gastric emptying. Incorporation of 10, 25, and 40 g lactulose into our standard meal in place of sucrose increased the rate of transit through the small intestine but did not significantly alter the rate of gastric emptying or the whole gut transit time. Total stool weight for 48 hr after ingestion of the meal was inversely related to whole gut transit time but not to small intestinal transit time suggesting that the tendency to develop diarrhea in response to a meal containing unabsorbable carbohydrate depends more on the lack of colonic accommodation than on the rate of small intestinal transit. Finally, there was no significant correlation between the measurements of small intestinal transit time after a drink of lactulose and the transit time of a meal in the same subjects.

Adult↗

A mechanochemical model of flagellar activity.

A theory is presented which quantitatively links the physical properties of a flagellum with parameters which characterize the chemical reactions responsible for deforming the flagellum. Realistic values for the wave parameters are predicted when order-of-magnitude values for the appropriate constants are used. The model may be useful in other fields where mechanochemical coupling occurs.

Flagella↗