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Biomedical subjects

J Thompson

Publications and source records attributed to J Thompson.

At least 541 records · Page 30Linked to original sources

Measurement of lung volumes during active and quiet sleep in infants.

The question of whether functional residual capacity (FRC) falls in infants during active sleep has been clouded by studies using different subject groups and techniques for measurements of lung volume and determination of sleep state. Twenty healthy full-term infants within the first week of life participated in the present study. Neurophysiological and behavioral criteria were used to define sleep state, and measurements of FRC were made using a specially constructed closed-circuit helium dilution system. Regularity of respiration was recorded using magnetometers on the chest and a modified respirator monitor. Results showed that no significant changes in FRC occurred, related either to sleep state or to regularity of respiration. In addition, we failed to detect any differences in FRC between the sexes.

Body Weight↗

The effects of high intensity training upon respiratory gas exchanges during fixed term maximal incremental exercise in man.

The response of respiratory gas exchanges to a 6 week high intensity training program was examined in 5 healthy males during fixed term maximal incremental treadmill exercise. Training was performed 3 d.wk-1 and consisted of a progressive series of repeated 15 sec and 30 sec maximal runs, and weight training exercises for the leg extensor muscles. Respiratory gases during the tests were continuously monitored using an on-line system. Muscle biopsy samples were obtained from the m. vastus lateralis before and after training for histochemical determination of fibre distribution based on myosin ATP-ase activity, and fibre cross-sectional area based on NADH-Tetrazolium Reductase activity. Training significantly increased the proportion of type IIa fibres (+5.9 +/- 2.0%, p less than 0.001) and decreased type I fibres (-6.3 +/- 2.0%, p less than 0.001), the distribution of type IIb fibres remained unchanged (+0.4 +/- 0.9%). Muscle cross-sectional area also showed a significant increase after training in type I (+318 +/- 215 microns 2, p less than 0.05), IIa (+652 +/- 207 microns 2, p less than 0.001) and IIb (+773 +/- 196 microns 2, p less than 0.001) fibres. During fixed term maximal incremental exercise the mean carbon dioxide output (VCO2) and mean respiratory exchange ratio (R = VCO2/VO2) were significantly increased (p less than 0.01) after training. The R-time relationship was at all times shifted to the left after training, being significantly (p less than 0.01) so over the final five min of exercise. No changes in mean exercise oxygen uptake (VO2), maximum oxygen uptake (VO2max) and maximum heart rate (FHRmax) were observed between tests.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

The effects of dietary manipulation upon the respiratory exchange ratio as a predictor of maximum oxygen uptake during fixed term maximal incremental exercise in man.

This study examined the effects of dietary manipulation upon the respiratory exchange ratio (R = VCO2/VO2) as a predictor of maximum oxygen uptake (VO2max). Seven healthy males performed fixed term maximal incremental treadmill exercise after an overnight fast on three separate occasions. The first test took place after the subjects had consumed their normal mixed diet (45 +/- 5% carbohydrate (CHO] for a period of three days. This test protocol was then repeated after three days of a low CHO diet (3 +/- 2% CHO), and again after three days of a high CHO diet (61 +/- 5% CHO). Respiratory gases were continuously monitored during each test using an on-line system. No significant changes in mean exercise oxygen uptake (VO2), VO2max or maximum functional heart rate (FHRmax) were found between tests. Mean exercise carbon dioxide output (VCO2) and R were significantly lower than normal after the low CHO diet (both p less than 0.001) and significantly higher than normal after the high CHO diet (both p less than 0.05). Moreover, compared with the normal CHO diet, the R-time relationship during exercise was at all times significantly (p less than 0.001) shifted to the right after the low CHO diet, and shifted to the left, being significantly so (p less than 0.05) over the final 5 min of exercise, after the high CHO diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

A noise-free molecular hybridization procedure for measuring RNA in cell lysates.

A solution hybridization technique was designed to measure RNA abundance in crude cell lysates and at the same time to maximize confidence that signals resulted from true molecular hybridization. Cell lysates were prepared in 5 M guanidine thiocyanate, then RNA molecules in the lysates were hybridized with two probes, a 32P-labeled RNA "label probe" which provided signal and an oligodeoxyribonucleotide "capture probe" containing a poly(dA) tail which provided a mechanism for selective purification. Ternary hybrids were "captured" on oligo(dT)-coated superparamagnetic beads through a readily reversible interaction with the poly(dA) of the capture probe. RNA did not bind to dT beads through poly(A) under the capture conditions used. Hybrids were purified through cycles of capture on and release from dT beads, with each cycle yielding a 100- to 1000-fold reduction in noise (unhybridized label probe) and a 50-90% recovery of signal (hybridized label probe). Noise was driven below detectable limits after three cycles of capture, thereby improving the sensitivity of measuring target RNA. As few as 15,000 target molecules, 15 fg of a 3-kb RNA, was detectable in the equivalent of 2 x 10(6) cells in concentrated cell lysates (10(8) cells/ml). Since hybridization with both probes was required in order to yield a signal, hybridization specificity could be adjusted with either or both probes. The greater specificity and lack of noise increased confidence that the signal was proportional to the amount of RNA of interest.

Guanidines↗

Allergenic and antigenic relationship between three species of storage mite and the house dust mite, Dermatophagoides pteronyssinus.

We have explored the antigenic and allergenic relationship between the house dust mite Dermatophagoides pteronyssinus and three species of storage mite, Glycyphagus destructor, Acarus siro, and Tyrophagus longior. Crossed immunoelectrophoresis demonstrated that all the mite extracts contained multiple antigens but that there was only limited cross-reactivity between the different species. Six sera were obtained from workers exposed to storage mites and with occupationally related lower respiratory tract symptoms. All workers had specific IgE to D. pteronyssinus and to one or more of the storage mites. The pattern of reactivity varied between the different sera, two responded primarily to D. pteronyssinus and A. siro and four sera to D. pteronyssinus and G. destructor. Only weak responses were observed to T. longior. RAST-inhibition and affinity-absorption experiments demonstrated that D. pteronyssinus had at least three groups of distinct allergenic determinants, determinants specific to D. pteronyssinus, determinants shared with A. siro, and determinants shared with G. destructor. Similarly, both A. siro and G. destructor have specific allergenic determinants and determinants shared with D. pteronyssinus. The findings demonstrate the complexity of the immunologic responses to the different mite species.

Allergens↗

Probes for quantitating subpicogram amounts of HIV-1 RNA by molecular hybridization.

A set of probes was designed for the quantitation of HIV-1 RNA in infected cells by a molecular hybridization procedure called reversible target capture. Reversible target capture is analogous to sandwich hybridization, except that the link between hybrid complexes and the affinity support was reversible, allowing for repeated capture of hybrids on, and release from, fresh affinity support. Repeated cycles of capture resulted in a high degree of purification of hybrids from unreacted probe, thereby greatly reducing assay noise and increasing assay sensitivity. Probes against the HIV-1 pol gene were chosen because their target sequences were highly conserved among HIV-1 isolates, while being divergent enough to provide discrimination from other human T cell tropic viruses. Subpicogram quantities of HIV-1 pol gene RNA were measured with signal:noise ratios of over 10. Hybridization signal increased with increasing target RNA with a proportionality constant of 1.

Cells, Cultured↗

Cesium-137, 134Cs and 110mAg in lambs grazing pasture in NE Scotland contaminated by Chernobyl fallout.

The decline in Cs radioisotope levels has been studied in tissues from lambs grazing lowland pasture. The lambs were slaughtered 18 and 115 d after contamination with Chernobyl fallout. During this time the Cs activity decreased to 13% of the initial amount in animals that had continued to graze contaminated pasture and to 3.5% in animals consuming uncontaminated feed. The 137Cs concentration in grass from the field grazed by the lambs decreased with a half-time of 22 d over the period 11-100 d after contamination. The amounts of Cs radionuclides removed from the pasture by the grazing animals amounted to only 0.01% of the total, the rest remaining in the soil, with over 40% in the upper 10 mm. Small amounts of 110mAg, found in grass, declined with a half-time of 8.9 d, and the radionuclide was found to accumulate in liver tissue.

Accidents↗

Alternative splicing of the neural cell adhesion molecule gene generates variant extracellular domain structure in skeletal muscle and brain.

Myotube mRNA isoforms of the neural cell adhesion molecule (N-CAM) contain a novel sequence block termed muscle-specific domain 1 (MSD1), which is inserted within the extracellular coding region. Here, we report a characterization of the genomic organization of MSD1 and its pattern of expression within cellular N-CAM RNA and polypeptide species. S1 nuclease protection analyses and sequence analysis of an N-CAM human genomic clone containing MSD1 sequences indicated that MSD1 is comprised of three discrete exons of 15, 48, and 42 bp, designated MSD1a, MSD1b, and MSD1c, respectively. Although the MSD1a exon was present in a small proportion of mRNAs from both brain and muscle cells, the entire MSD1 sequence occurred predominantly in mRNAs from differentiated myotube cells. In addition, antiserum raised to a synthetic, MSD1b-encoded peptide sequence was found to stain the cell surface of human skeletal myotubes in culture, whereas myoblasts, fibroblasts, and neural cells were negative. MSD1a, MSD1b, and MSD1c sequences thus arise collectively in N-CAM mRNA and polypeptide isoforms as a result of muscle tissue-specific and developmentally regulated alternative mRNA splicing events. In addition, the occurrence of brain and muscle mRNAs containing only MSD1a indicate that alternative splicing may occur within the MSD region itself to generate further diversity.

Amino Acid Sequence↗

Double labelling of tissue combining tritiated thymidine autoradiography with immunodetection of bromodeoxyuridine: the autoradiographic significance of inhibition of thymidine incorporation into DNA by bromodeoxyuridine given simultaneously.

We describe a reproducible method for combining tritiated thymidine ([H]TdR) autoradiography with immunoperoxidase detection of bromodeoxyuridine (BrdU) in paraffin-embedded tissues. The technique has been used to examine, in mouse tongue epithelium, the inhibition of incorporation into DNA of [3H]TdR by a simultaneous injection of BrdU in the doses that both compounds are likely to be used in cell proliferation studies. The significance that this inhibition has on prolongation of autoradiograph exposure times, to ensure that all cells that incorporate [3H]TdR are scored as positive, in particular the most lightly labelled cells, has been quantified. The inhibition of uptake into DNA of [3H]TdR from 0.23 to 1.85 MBq (6.25 to 50 mu Ci) per animal, produced by a simultaneous injection of 2.5 mg BrdU shows a linear, dose-dependent relationship. Provided the injected dose (in mu Ci per animal) multiplied by the autoradiographic exposure time (in days) is greater than a value of 700, then all cells that are labelled after incorporation of [3H]TdR alone are also labelled after simultaneous double labelling, despite the latter producing a lower average grain count.

Animals↗

School health services in the United States: a view from the United Kingdom.

Despite sharing both a common heritage and a common language, differences exist between life in the United States and the United Kingdom. During spring 1988, the author, who formerly served as editor for the Health Visitor, published by the Health Visitor's Association, completed a six-week study tour of school health services in the United States. In this article, she gives her impressions from the experience, including observations regarding salary, working conditions, professional preparation, and professional recognition. She concludes by identifying several poignant questions to clarify the school nurse's role and to set priorities for the optimal use of time and expertise.

Attitude of Health Personnel↗

Respiratory control in infants at increased risk for sudden infant death syndrome.

There is much debate relating to possible abnormalities in respiratory control mechanisms in infants considered at increased risk for sudden infant death syndrome (SIDS). The P0.1 occlusion technique was used to assess the central respiratory response to hyperoxic hypercapnia during quiet sleep in 21 normal infants, 13 siblings of SIDS victims, and 17 infants with apparent life threatening events. The slope of P0.1 plotted against carbon dioxide concentration increased exponentially with age, independent of body weight in each group. Birth weight has a significant effect on slope with a lower weight predisposing to a lower slope. Siblings as a group had a significantly lower slope at any given age than normal infants, whereas the infants who had had apparent life threatening events were not significantly different from the controls. As intragroup variation in both siblings and control groups greatly exceeded the significant intergroup differences observed, the technique cannot identify individual infants as belonging to one or other group.

Age Factors↗

Coronary pressure-flow relations in hypertensive left ventricular hypertrophy. Comparison of intact autoregulation with physiological and pharmacological vasodilation in the dog.

Coronary pressure-flow relations during autoregulated and vasodilated flow states were compared between eight dogs with renovascular hypertension and left ventricular hypertrophy and 12 normal dogs. Each relation was constructed from serial steady-state measurements of end-diastolic coronary pressure and flow during perfusion of the circumflex artery by an extracorporeal circuit at controlled diastolic pressures of 20-200 mm Hg. Autoregulated pressure-flow relations were compared at three levels of myocardial oxygen demand: resting, high (dobutamine 10 micrograms/kg/min), and low (propranolol 2.5 micrograms/kg/min). Autoregulatory capacity was assessed by calculation of closed-loop flow gain. At each level of myocardial oxygen demand, the lower limit of autoregulation occurred at higher perfusion pressures in the hypertrophy group (rest 65 +/- 3, high 92 +/- 4, low 66 +/- 4 mm Hg) than in the normal group (rest 53 +/- 2, p less than 0.05; high 75 +/- 5, p less than 0.05; low 51 +/- 3 mm Hg) (p less than 0.05). Maximum autoregulatory gain was similar in the normal and hypertrophy groups during resting and low myocardial oxygen demand but was reduced in the hypertrophy group during dobutamine studies. When coronary flow decreased below the lower limit of autoregulation, systolic shortening was reduced in both normal and hypertrophy groups. However, as the autoregulatory limits were at higher pressures in the hypertrophy group, shortening in this group deteriorated at perfusion pressures that did not affect the normal heart. Coronary pressure-flow relations during physiological (peak hyperemia after 15-second flow occlusion) and pharmacologica (intracoronary adenosine 400 micrograms/min) vasodilation was curvilinear and fitted by quadratic regression. During hyperemic vasodilation, maximal conductance per unit mass of myocardium was less in the hypertrophy group over a wide range of perfusion pressures. At a diastolic perfusion pressure of 80 mm Hg, maximum conductance was 4.6 +/- 0.5 ml/min/100 g/mm Hg in the normal group and 3.4 +/- 0.4 ml/min/100 g/mm Hg (p less than 0.05) in the hypertrophy group. Intracoronary adenosine elicited further vasodilation in both groups, but maximum conductance remained less in the hypertrophy group (8.5 +/- 1.7 ml/min/100 g/mm Hg at a perfusion pressure of 80 mm Hg) than in the normal group (13.5 +/- 2.0 ml/min/100 g/mm Hg) (p less than 0.05). Maximal coronary flow reserve is reduced in left ventricular hypertrophy, with a consequent shift of the lower limit of autoregulation to higher perfusion pressures. Thus, as coronary perfusion pressure is decreased, coronary flow and myocardial shortening become impaired at higher

Animals↗

The role of angiotensin-II in progressive diabetic glomerulopathy in the rat.

The potential pathogenic role of angiotensin-II (AII) in early progressive diabetic and renal ablation-induced glomerulosclerosis was explored and compared in the Sprague-Dawley (SD) rat and the mongrel dog. Male SD rats were divided into control and streptozotocin-treated (65 mg/kg, iv) groups. Unilateral surgical nephrectomy (Nx) was performed in half of each group. Enalapril (E; 50 mg/liter in the drinking water) was administered to half of each subgroup. Enalapril (high E; 250 mg/liter) was given to another 13 streptozotocin rats. All diabetic rats were treated with sc NPH insulin (4 U/day), and blood glucose was 520 +/- 124 mg/dl (mean +/- SD). Microalbuminuria was measured by RIA in 24-h urine collections; wet kidney weights were compared. [125I]AII binding assays were performed on isolated glomeruli. In control rats the high affinity binding dissociation constant (Kd) was 0.59 +/- 0.15 nM (n = 26; mean +/- SD) and receptor number (Ro) was 732 +/- 195 fmol/mg glomerular protein. At 3 weeks, the diabetic glomerular AII receptor Kd was 0.38 +/- 0.07 nM (n = 6; P less than 0.02 vs. control) and Ro was 784 +/- 97 fmol/mg protein (P = NS vs. control); diabetic high E Kd was 0.39 +/- 0.06 nM (n = 6; P less than 0.02 vs. control), and Ro was 873 +/- 105 fmol/mg protein (P = NS vs. diabetes without E). By 10 weeks, a Kd of 0.49 +/- 0.12 nM (n = 32; P less than 0.01 vs. control) and a Ro of 780 +/- 174 fmol/mg protein (P = NS vs. control) were observed when all of the diabetic group data were pooled. Neither Nx nor low or high dose E altered Ro. This is evidence that intraglomerular AII levels are normal or reduced after diabetes, Nx, or both. In the diabetic group, low dose E partially prevented, and high E abolished, Nx-enhanced microalbuminuria and renal hypertrophy. In nine pancreatectomized insulin-treated mongrel dogs over a 12- to 24-month period, despite moderately poor glucose control (300 +/- 75 mg/dl) and combined unilateral Nx in five dogs (12 months), elevated microalbuminuria was not observed. [125I]AII binding to isolated normal and diabetic dog glomeruli revealed the Kd to be of low affinity (1.86 +/- 0.28 to 13.80 +/- 1.88 nM), identifying the presence of type B receptors. Hence, the SD rat and mongrel dog differ in susceptibility to glomerular AII receptor type and progressive diabetic glomerulopathy.(ABSTRACT TRUNCATED AT 400 WORDS)

Albuminuria↗

Glomerular epithelial detachment, not reduced charge density, correlates with proteinuria in adriamycin and puromycin nephrosis.

To identify the structural change coincident with increased glomerular permeability in both adriamycin (ADR) and puromycin-aminonucleoside (PAN) nephrosis, we explored the temporal correlation between developing proteinuria, the reduction in glomerular polyanions, and the detachment of epithelium from glomerular basement membrane (GBM). Sprague-Dawley rats received a single tail-vein injection of PAN (150 mg/kg), ADR (7.5 mg/kg) or saline. Nephrotic-range proteinuria appeared between days 2 to 5 in the PAN-treated and days 5 to 10 in the ADR-treated rats. The GBM heparan sulfate charge density and epithelial membrane sialic acid (SA) content were determined before, during and after the rise in proteinuria. Polyethyleneimine was used to detect heparan sulfate and the number staining of renal cortical slices was used to detect heparin sulfate and the number of sites/microns GBM in the lamina rara externa were counted on electron micrographs (magnification x60,000). Controls had a regular distribution of polyethyleneimine 20.19 +/- 1.72 sites/microns (X + SD). In ADR rats, the polyethyleneimine density decreased by day 5, 18.61 +/- 1.79 sites/microns (p less than 0.05) which persisted to day 15, 17.38 +/- 1.27 sites/microns (p less than 0.02). PAN rats, by day 1, had significant reduction, 14.94 +/- 1.47 sites/microns (p less than 0.05), which persisted to day 20. The total membrane-bound SA content of isolated glomeruli was analyzed with a modified Warren's method. The SA content in control glomeruli was 46.8 +/- 8.0 nmol/mg glomerular protein (X +/- SD). In ADR rats, there was significant decrease in SA content to 84 +/- 3% of control (p less than 0.01) at day 15. In PAN rats, by day 2, the SA content was decreased to 73 +/- 18% of control (p less than 0.05). In both models, scanning and transmission electron microscopy revealed epithelial foot process fusion, loss of slit diaphragms, and vacuolization before increased proteinuria. Epithelial detachment from the GBM and rupture of vacuoles occurred coincidently with rapid development of nephrotic proteinuria in both models. In summary, reduced GBM heparan sulfate and epithelial SA content do not correlate with the onset of altered glomerular permeability, whereas epithelial detachment is coincident with the development of massive proteinuria in both ADR and PAN nephrosis.

Animals↗