In vivo imaging of nitroxide-free-radical clearance in the rat, using radiofrequency longitudinally detected ESR imaging.
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Biomedical subjects
Publications and source records attributed to M A Foster.
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Three imaging methods, ultrasound imaging (UI), computer-assisted axial tomography (CAT) and magnetic resonance imaging (MRI), are widely used in medicine. Their application to the assessment of body composition in nutrition research is still being explored and developed. Ultrasound imaging yields poor image quality but, because it is cheap and safe, deserves further exploration. Both CAT and MRI can produce images with good discrimination among bone, muscle and adipose tissue. Movement artifacts tend to be more serious in MRI than in CAT due to the longer imaging time. On the other hand, the X-ray exposure in CAT is likely to limit its use in human nutrition research. Repeated measurements of tissue volumes by CAT and MRI give similar CV. In both CAT and MRI, intra-abdominal adipose tissue presents greater problems of measurement than subcutaneous adipose tissue. Validation studies with 77-kg pigs of MRI, using 13 slices, predicted total body lipid with residual standard deviation of 1.9%. In validating any of these methods, account should be take of the extent to which the information they give can augment that given by more simple measures like age and weight.
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In vivo quantification of adipose tissue with magnetic resonance imaging (MRI) was validated with pigs. Thirteen transaxial MRI sections were collected, at intervals proportional to body length, from each pig, which was then killed, frozen, and sliced at the locations of the MRI sections. Adipose-tissue quantities were determined by dissecting each slice, and lipid contents of the dissected slices and of the tissue segments between slices were measured. Compared with dissection, MRI underestimated abdominal percent adipose tissue and overestimated cervical percent adipose tissue by less than 6%. When all 13 sections were used, MRI closely predicted percent lipid and dissected percent adipose tissue with small residual SDs (RSD = 1.9 and 2.1, respectively), which increased only slightly if two sections (4, upper thorax and 8, upper abdomen) were used (RSD = 2.3 and 2.6, respectively). In conclusion MRI accurately quantifies adipose tissue in vivo, matching values produced by dissection and chemical analysis.
In the cofermentation of glycerol with a sugar by Lactobacillus brevis and Lactobacillus buchneri, a 1,3-propanediol:NAD+ oxidoreductase provides an additional method of NADH disposal. The enzyme has been purified from both L. brevis B22 and L. buchneri B190 and found to have properties very similar to those reported for the enzyme from Klebsiella pneumoniae. The enzymes required Mn2+ and are probably octamers with a molecular mass of 350 kDa. Although not absolutely specific for 1,3-propanediol when tested as dehydrogenases, the enzymes have less than 10% activity with glycerol, ethanol, and 1,2-propanediol. These properties contrast sharply with those of a protein isolated from another Lactobacillus species (L. reuteri) that ferments glycerol with glucose and previously designated a 1,3-propanediol dehydrogenase.
The simultaneous fermentation of glycerol and sugar by lactobacillus brevis B22 and Lactobacillus buchneri B190 increases both the growth rate and total growth. The reduction of glycerol to 1,3-propanediol by the lactobacilli was found to influence the metabolism of the sugar cofermented by channelling some of the intermediate metabolites (e.g., pyruvate) towards NADH-producing (rather than NADH-consuming) reactions. Ultimately, the absolute requirement for NADH to prevent the accumulation of 3-hydroxypropionaldehyde leads to a novel lactate-glycerol cofermentation. As a result, additional ATP can be made not only by (i) converting pyruvate to acetate via acetyl phosphate rather than to the ethanol usually found and (ii) oxidizing part of the intermediate pyruvate to acetate instead of the usual reduction to lactate but also by (iii) reoxidation of accumulated lactate to acetate via pyruvate. The conversion of lactate to pyruvate is probably catalyzed by NAD-independent lactate dehydrogenases that are found only in the cultures oxidizing lactate and producing 1,3-propanediol, suggesting a correlation between the expression of these enzymes and a raised intracellular NAD/NADH ratio. The enzymes metabolizing glycerol (glycerol dehydratase and 1,3-propanediol dehydrogenase) were expressed in concert without necessary induction by added glycerol, although their expression may also be influenced by the intracellular NAD/NADH ratio set by the different carbohydrates fermented.
Taste sensitivity to five glutamate salts (sodium glutamate, potassium glutamate, ammonium glutamate, calcium diglutamate, and magnesium diglutamate) were determined in sixteen young (mean age 25.58 years) and eighteen elderly (mean age 86.89 years) subjects. The effect of inosine 5'-monophosphate (IMP) and ions on taste perception of glutamate compounds was also investigated. The detection thresholds for glutamate salts were 5.04 times higher in elderly subjects than in young subjects; the recognition thresholds were 3.84 times higher. For young subjects, 0.1 mM IMP lowered detection and recognition thresholds for all 5 salts. A stronger concentration of IMP (1 mM) had this effect in both young and elderly groups. Elderly subjects perceived suprathreshold concentrations as less intense than young subjects. Chloride and acetate salts of sodium, potassium, and calcium reduced the detection and recognition thresholds of L-glutamic acid but had no effect sodium glutamate thresholds.
Mammogenesis and lactation were induced in five multiparous, non-pregnant goats by treatment with oestrogen and progesterone for 11 d, followed by dexamethasone for 3 d. Reserpine was administered during the last 5 d. All five goats lactated, although milk yield was less than had been achieved in previous natural lactations. Mammary development was assessed in vivo, using magnetic resonance imaging. Although parenchyma volume increased by more than 6-fold overall, only 25% of this increase occurred during steroid treatment. Most development took place after the cessation of treatment, when milking commenced. Maximum size was not achieved until week 8 of the induced lactation, and was only 70% of normal parenchyma volume. After 18 weeks lactation the activities of three key milk synthetic enzymes were very similar to values previously found in natural lactations, and secretion efficiency (milk production per unit volume of parenchyma) was also similar to that of natural lactations. We conclude that the lower than normal milk yields were associated with incomplete proliferation of mammary tissue, rather than inadequate differentiation of individual secretory cells.
Four goats were studied from the end of their second lactation. One mammary gland of each goat was dried off just prior to the start of the third gestation, whilst the other gland was milked throughout gestation, with no dry period. At the end of gestation the continuously milked gland was significantly smaller than the gland that had been allowed a dry period. However, this difference did not persist beyond parturition and there was no significant difference between the milk yields of the two glands in the next lactation, although the continuously milked gland tended to have the higher yield. At 18 weeks of lactation, mammary parenchyma weight and secretory cell number were significantly greater in the continuously milked gland, but mammary enzyme activities did not differ between the two glands.
Total and subcutaneous adipose tissue in seven lean and seven obese women were quantified using magnetic resonance imaging (MRI). The distributions of adipose tissues along the body were closely correlated: subcutaneous with total, both within and between lean and obese groups. Lean women had proportionally less adipose tissue in the lower thorax and upper abdomen than did obese women. Reducing the number of MRI scans from 17 to 4 did not increase the residual SD of predicted body adipose tissue (2.9 percent) when body density was used as the reference measure. MRI gave an estimate of total-body adipose tissue significantly closer to the value for fat percent produced when the results from five other techniques (skinfold thickness, underwater weighing, 40K whole-body counting, isotopic water dilution, and tetrapolar bioelectrical impedance) were averaged than when any other technique was used alone. MRI-determined percent body adipose tissue in women is close to, and proportional to, estimates derived by underwater weighing.
Red blood cell proton relaxation times T1 and T2 were measured in samples from chronic alcoholic patients abstinent for varying periods from 1 week to over 6 months. T1 and T2 were elevated in the early stages of abstinence and declined to the values of controls after 8 weeks. Changes in the water content of the red blood cells and the mean corpuscular volume paralleled these changes but were more closely associated with T2. It is suggested that T1 and T2 may reflect different aspects in water content and free-to-bound ratio of water. The significance of these findings is discussed in the context of changes previously observed in the brains of alcoholic patients, and in rats fed a diet supplemented with alcohol for 6 months.
The clinical performance of three posterior composites and two amalgams was evaluated over 3 years. Three hundred and nine composites and 200 amalgams were reviewed at the 3-year recall. The following parameters were assessed: fractures, gingival condition, interproximal contacts, marginal adaptation, staining and colour match. Although there was no significant difference in the overall failure rate between the two types of material, there was a significantly higher incidence of fracture in the amalgam restorations. The amalgam failures were exclusively mechanical, whereas those affecting composites also included failures that were more biological in nature. A significantly higher proportion of the Class II composites failed to achieve an adequate approximal contact, but this was not reflected in a more adverse gingival response. Colour match, surface and marginal stain varied significantly within the composite group and appeared to be material-related. Overall, the materials Occlusion and P-30 were considered to be performing adequately. The clinical handling, colour match and radiopacity of Clearfil Posterior were considered to be unsatisfactory.
Pellets of HeLa from suspension cultured cells in isotonic medium (300 mosmolar) were introduced into a Bruker CXP100 NMR spectrophotometer at 80 mHz within 5 min of the start of centrifugation. T1 and T2 times were measured within a total elapsed time of 20-25 min at 80 mHz and 37 degrees C, and averaged 1430 msec and 120 msec, respectively. Extrapolation to zero extracellular space gave a corrected T1 of 1370 msec. For cells collected after 10 min in hypotonic medium (down to 30 mosmolar) increased proton density correlated well with increased cell water content, but relaxation times did not rise in proportion to that predicted for the entry of "bulk" water (T1 of 4700 msec), except when swelling approached lysis point. Cells partially dehydrated by 10 min in hypertonic medium of up to 1500 mosmolar have also been analyzed, but once again the shortening of T1 was not proportional to the loss of "free" (bulk phase) water. At the upper limit of hypertonic treatment, lacunae or vacuoles of a watery nature separated within the cytomatrix, preventing maximum dehydration. The relationship of cell water to T1 is complex over the whole range of tonicity that HeLa S-3 cells tolerate. The data indicate, however, that hypotonically induced water probably has an average T1 time considerably lower than bulk phase water. In contrast, raising the total extracellular volume with medium had precisely the predicted effect on T1 time, further strengthening the case that water taken up by cell acquires a shorter T1 time. Cells adapting to hypotonic conditions oscillated in size and water content over 2-3 hr before returning to near their initial volume. Under these circumstances, T1 oscillated in the same way but with a reduced amplitude, consistent with the above findings.
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Washed rubber particles isolated from stem homogenates of Parthenium argentatum Gray by ultracentrifugation and gel filtration on columns of LKB Ultrogel AcA34 contain rubber transferase which catalyzes the polymerization of isopentenyl pyrophosphate into rubber polymer. The polymerization reaction requires Mg(2+) isopentenyl pyrophosphate, and an allylic pyrophosphate. The K(m) values for Mg(2+), isopentenyl pyrophosphate, and dimethylallyl pyrophosphate were 5.2 x 10(-4) molar, 8.3 x 10(-5) molar, and 9.6 x 10(-5) molar, respectively. The molecular characteristics of the rubber polymer synthesized from [(14)C]isopentenyl pyrophosphate were examined by gel permeation chromatography on three linear columns of 1 x 10(6) to 500 Angstroms Ultrastyragel in a Waters 150C Gel Permeation Chromatograph. The peak molecular weight of the radioactive polymer increased from 70,000 in 15 minutes to 750,000 in 3 hours. The weight average molecular weight of the polymer synthesized over a 3 hour period was 1.17 x 10(6) compared to 1.49 x 10(6) for the natural rubber polymer extracted from the rubber particles. Over 90% of the in vitro formation of the rubber polymer was de novo from dimethylallyl pyrophosphate and isopentenyl pyrophosphate. Treatment of the washed rubber particles with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate solubilized the rubber transferase. The solubilized enzyme(s) catalyzed the polymerization of isopentenyl pyrophosphate into rubber polymer with a peak molecular weight of 1 x 10(5) after 3 hours of incubation with Mg(2+) and dimethylallyl pyrophosphate. The data support the conclusion that the soluble preparation of rubber transferase is capable of catalyzing the formation of a high molecular weight rubber polymer from an allylic pyrophosphate initiator and isopentenyl pyrophosphate monomer.
The volumes and spin-lattice (T1) relaxation times of breast tissues and parenchymal water content were measured non-invasively by magnetic resonance imaging (MRI) in eight healthy women during four to eight consecutive menstrual cycles. Total breast volume, and parenchymal volume, T1 relaxation time and water content were lowest between days 6 and 15. Between days 16 and 28, parenchymal volume, T1 relaxation time and water content rose sharply by 38.9%, 15.1% and 24.5%, respectively, and peaked after day 25. Within 5 days of the onset of menses, parenchymal volume fell sharply by 30.3%, while water content declined by 17.5%. Rising parenchymal volume in the second half of the menstrual cycle is not solely due to increased tissue water content and provides in vivo evidence for both growth and increased tissue fluid at this time.
A technique is described for the in-vivo determination of mammary gland size and gross composition in goats by using nuclear magnetic resonance imaging (MRI). The volume of test objects determined with MRI had an error of +0.4 +/- 1.6% of the actual volume. In lactating goats the in-vivo MRI estimate of mammary parenchymal volume was significantly greater than, but highly significantly correlated with, the weight of parenchyma determined post mortem (for the whole udder, r = 0.88, P less than 0.001; for individual glands, r = 0.85, P less than 0.001). MRI-determined estimates of the volume of fluid within the mammary gland were within 1.2% of the volume of milk removed from the udders after imaging. The spin-lattice (T1) relaxation time of the whole udder correlated closely with the volume of fluid within the udder. The T1 relaxation time of parenchymal tissue measured in vivo did not differ significantly from that determined immediately after post-mortem excision.
Mammary development and regression were measured in goats in vivo using magnetic resonance imaging (MRI). Measurements were made during the first and second cycles of pregnancy, lactation and involution. In primiparous goats, and exponential pattern of growth was evident during gestation and for the first 2 weeks of lactation. Parenchyma volume correlated significantly with milk yield across goats during early lactation, and across stage of lactation within goats. Milking was discontinued in Week 26 of the first lactation. Involution was characterized by an initial accumulation of fluid (over 2 days) followed by reabsorption; parenchyma volume did not decrease significantly until the 3rd week of involution, which was also the time at which these goats were mated to start their second gestation. Their udders still contained significant quantities of fluid (40-60% of the gross volume), but parenchyma volume was also greater (by 4.7-fold) than in goats beginning their first gestation. By Week 15 of gestation there was no longer a parity difference in parenchyma; the udders of first-gestation goats had grown significantly, but those of second-gestation goats had not. Conversely, between gestation Week 15 and lactation Week 2 mammary growth was significantly more rapid in the second cycle, such that the udder was larger at the start of the second lactation.