Self referral to an accident and emergency department.
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Biomedical subjects
Publications and source records attributed to J Green.
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1. The roles of the three protein components of soluble methane mono-oxygenase were investigated by the use of rapid-reaction techniques. The transfer of electrons through the enzyme complex from NADH to methane/O2 was also investigated. 2. Electron transfer from protein C, the reductase component, to protein A, the hydroxylase component, was demonstrated. Protein C was shown to undergo a three-electron--one-electron catalytic cycle. The interaction of protein C with NADH was investigated. Reduction of protein C was shown to be rapid, and a charge-transfer interaction between reduced FAD and NAD+ was observed; this intermediate was also found in static titration experiments. Thus the binding of NADH, the reduction of protein C and the intramolecular transfer of electrons through protein C were shown to be much more rapid than the turnover rate of methane mono-oxygenase. 3. The rate of transfer of electrons from protein C to protein A was shown to be lower than the reduction of protein C but higher than the turnover rate of methane mono-oxygenase. Association of the proteins was not rate-limiting. The amount of protein A present in the system had a small effect on the rate of reduction of protein C, indicating some co-operativity between the two proteins. 4. Protein B was shown to prevent electron transfer between protein C and protein A in the absence of methane. On addition of saturating concentrations of methane electron transfer was restored. With saturating concentrations of methane and O2 the observed rate constant for the conversion of methane into methanol was 0.26 s-1 at 18 degrees C. 5. By the use of [2H4]methane it was demonstrated that C-H-bond breakage is likely to be the rate-limiting step in the conversion of methane into methanol.
Washing thylakoid membranes with 1 M LiCl causes the release of the beta subunit from the chloroplast energy transducing complex (CF1.CF0) in spinach chloroplasts. This protein purifies by size exclusion chromatography as a 180-kDa aggregate and, thus, is probably composed of a trimer of beta polypeptides. The purified aggregate binds ADP to a high and a low affinity site with dissociation constants of 15 and 202 microM, respectively. Mg2+ is required for ADP to bind to both sites. Manganese binds to the protein in a cooperative manner to at least two sites with high affinity. The beta subunit preparation catalyzes Mg2+-dependent ATP hydrolysis at rates which are comparable to other subunit-deficient CF1 preparations and is increased by treatments known to activate the Mg2+-ATPase activity of CF1. However, Ca2+ is not an effective cofactor for this reaction and treatments which activate the Ca2+-ATPase of CF1 are either ineffective or inhibitory.
Measurements of cell volume changes, free cytosolic Ca2+ concentration [( Ca2+]i) with Fura 2 and cell membrane potential with 3,3'-dipropylthiodicarbocyanine iodide were used to study the effect of cell volume change on Ca2+ influx and the membrane potential of the osteoblastic osteosarcoma cell line, UMR-106-01. Swelling the cells by hypo-osmotic stress was followed by reduction in cell volume which was markedly impaired by removal of medium Ca2+. Accordingly, cell swelling resulted in [Ca2+]i increase only in the presence of medium Ca2+. The cell swelling-activated Ca2+ entry pathway was active at resting membrane potentials, and Ca2+ influx through this pathway markedly increased upon cell hyperpolarization. A linear relationship between Ca2+ entry and the potential across the plasma membrane was observed. Thus, the volume-activated Ca2+ permeating pathway in UMR-106-01 cells has conductive properties. These pathways do not spontaneously inactivate with time when the cells are not allowed to volume regulate. The pathway can be blocked by micromolar concentrations of nicardipine and La3+ but display very low sensitivity to diltiazem and verapamil. Activation of the volume-sensitive, Ca2+ permeating pathway was not dependent on an increase in [Ca2+]i. Likewise, activation of the pathway was independent of a change in membrane potential between -85 and -3 mV. The increase in [Ca2+]i resulted in hyperpolarization of the cells, probably due to activation of Ca2+-activated K+ channels. The volume-sensitive pathways were partially active under isotonic conditions. Their activity was inhibited by cell shrinkage and increased by cell swelling. The pathways were sensitive to small changes in cell volume, particularly around a medium osmolarity of 310 mosM.
Measurements of free cystolic Ca2+ ([Ca2+]i) and Ba2+ ([Ba2+]i) concentrations with Fura 2 were used to identify and characterize the properties of a depolarization-activated Ca2+ and Ba2+ entry in the plasma membrane of osteoblast-like cells. The presence of this pathway was demonstrated in two osteoblastic cell lines, UMR-106 and MC3T3-E1 and osteoblasts isolated from rat long bone and rat neonatal calvariae. Subsequent characterization of the pathway was performed in the osteosarcoma cell line UMR-106. Depolarization of the cells with high medium K+ was followed by an increase in [Ca2+]i which was dependent on medium Ca2+. Ba2+ ions depolarized the cells and were transported by this pathway. Mg2+ ions interfered with Ca2+ and Ba2+ entry. At 140 mM KCl and 1 mM MgCl2, the pathway could be saturated with Ca2+ or Ba2+. The apparent affinity for Ca2+ was 0.78 mM and for Ba2+ 1.82 mM. Ca2+ or Ba2+ entry into the cells was blocked by low concentrations of nicardipine, diltiazem, verapamil, and La3+. In the absence of an increase in [Ca2+]i or [Ba2+]i, the pathway inactivated within about 5 min after depolarization. When [Ca2+]i or [Ba2+]i was allowed to increase, the pathway inactivated within about 20 s. These properties suggest that Ca2+ and Ba2+ entry are mediated by an L-type, depolarization-activated Ca2+ channel in osteoblasts. The activity of these channels changes little with an increase or decrease in cell volume. Thus, it is concluded that these pathways do not provide the Ca2+ entry pathway required for initiation of volume decrease by osteoblasts.
Paired scrapes and biopsies from 100 women attending a routine colposcopy clinic were examined by dot blot DNA hybridisation for infection with human papillomavirus (HPV) types 6/11, 16, 18, and 31; 51% of the scrapes and 50% of the biopsies were positive for HPV infection. Scrapes detected more HPV 18 (10% vs. 2%, P = less than 0.05) and HPV 31 (7% vs. 3%, not significant) than did the biopsies, but biopsies detected more HPV 16 (42% vs. 33%, not significant). Comparison of the results for each patient revealed that the correlation between scrapes and biopsies was not very close: only 34 patients were HPV-positive by both sampling methods, whereas 67 were positive if the results were combined. This report discusses the implications of these findings.
The factors responsible for meal initiation in man are not known. Recently, a small, transient decline in blood glucose was found to precede meals in freely feeding rats (1). To determine whether a similar decline in blood glucose precedes meals in humans, we measured plasma glucose concentration before and after requests for lunch on 4 different days in each of 7 subjects living in a setting that eliminated temporal cues and most or all external prompts for eating. Under these conditions, subjects could request any meal at any time by naming it and specifying its content. Meals were promptly prepared and served. Blood was sampled at randomized intervals averaging 20 minutes. Blood samples from 120 minutes preceding the subject's request for lunch to 90 minutes after the request were analyzed for glucose by the hexokinase method. The mean plasma glucose concentration did not change significantly from about 1 hour before lunch request to 20-30 minutes after the request. A transient decline in glucose similar to that reported in the rat was observed prior to only 1 of the 28 lunches. Given the ease of obtaining food and the absence of the usual temporal and psychosocial cues for eating, the results are not consistent with the hypothesis that meal requests in humans are triggered by a transient decline of plasma glucose concentration. The hypothesis is not disproven, however, since the intersample intervals of about 20 minutes may have failed to detect decreases of blood glucose that were smaller or of shorter duration than those of the rat.
The history of counselling in HIV infection and AIDS is reviewed and the stages in the development of the area are set out. The way in which the area has developed in the West is related to local circumstances and the need for the development of models suited to local circumstances elsewhere in the world is stressed. There are many areas of of HIV/AIDS counselling where considerable uncertainty about the right approach remains, for instance with injecting drug users and with HIV-infected pregnant women. There is a great and largely unmet need for further research in the area and some of the questions which need to be addressed are set out.
The mechanism of activation of Na+/H+ exchanger by various stimuli was studied in the human epidermoid carcinoma cell line A431 and in peripheral blood mononuclear cells (PBM). Intracellular pH (pHi) was measured by using the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. Stimulation of A431 cells by epidermal growth factor (EGF), bradykinin (BK), phorbol-12-myristate 13-acetate (PMA), and osmotic shrinkage resulted in exchanger activation. In PBM, activation of Na+/H+ exchanger was induced by concanavalin A (Con A) and phytohemagglutinin (PHA), as well as PMA and osmotic shrinkage. Inhibition of protein kinase C inhibited only PMA-stimulated exchanger activation in both cell types. When osmotic shrinkage was applied after exposure of the cells to any agonist, augmentation of exchanger activation by osmotic stress was observed. These findings suggest that various stimuli activate Na+/H+ exchanger through different mechanisms. Kinetic analysis demonstrated that activation of the exchanger by any type of stimulus resulted in modification of the apparent affinities for intracellular H+ (H+i) and intracellular Na+ (Na+i) in opposite directions. While there is an increased apparent affinity for H+i, the apparent affinity for Na+i decreases. This finding suggests that in A431 cells this phenomenon serves as a common mechanism for activation of Na+/H+ exchanger by different stimuli.
The relationship between verbal ability and aggressive behavior in 53 physically abused children in a residential treatment center was examined. Characteristics differentiating aggressive children from their less aggressive peers were identified. Although general verbal ability was similar in aggressive and nonaggressive abused children, reading and expressive language deficits were more prevalent in the highly aggressive children. Highly aggressive children were significantly more deficient in reading ability and more often required speech-language services. Implications of these findings for treatment and future research are discussed.
The size of an adenoid is generally considered to be a significant factor in the causation of Eustachian tube dysfunction and glue ear in children. We studied 120 consecutive cases of children undergoing adenoidectomy. Our aim was to assess the accuracy of the digital and mirror examinations in identifying adenoid size. Two surgeons of 1 and 2 years' experience in the speciality were found to have a statistically significant correlation between mirror examination and the amount of adenoid removed at operation. One surgeon of 4 years' experience was found to have a higher degree of correlation for both mirror and digital examinations.
Two stable clusters of parent-rated temperamental traits, a "drive" and a "regulatory" cluster, are identified in the first six months of life. The relationship among these clusters and neonatal behavior, measured on the Brazelton Neonatal Behavior Assessment Scale, is considered, and the nature of the clusters compared to the results of other studies with similar methodology. Implications of these results for an interactional theory of early development are discussed.
Fifteen semen specimens from 10 men with intrameatal penile warts attending a genitourinary clinic were tested by Southern blot hybridisation for the presence of human papillomavirus (HPV) DNA. Five specimens were positive for HPV types 6/11. This observation may have implications for screening of semen used for artificial insemination by donor.
The naturally occurring prostaglandins (PGs) were studied with respect to their abilities to change free cytosolic Ca2+ concentrations ([Ca2+]i), adenosine 3',5'-cyclic monophosphate (cAMP) levels, and cell proliferation in the osteoblastic cell line, UMR-106-01, and primary cultures of osteoblasts prepared from neonatal rat calvariae. All PGs tested stimulated an increase in [Ca2+]i, which was mainly due to Ca2+ release from intracellular stores. Measurements of the 50% effective concentration for the different PGs show that the potency ranking for PG-evoked [Ca2+]i increase in these cells is F2 alpha greater than D2 much greater than E2 greater than TxB2 greater than E1 greater than I2 much greater than A2. The PGs also increase cAMP levels in osteoblasts. At the highest concentrations tested (10-25 microM), dose-response saturation of cAMP production was observed only by PGE2 and PGE1. The potency rank for PG-stimulated cAMP increase was E2 greater than E1 much greater than A2 greater than I2 greater than F2 alpha greater than D2 greater than TxB2. Measurements of the effect of the PGs on thymidine uptake showed that low concentrations of PGF2 alpha and PGD2 had either no effect or stimulated proliferation of osteoblast-like cells. Relatively low concentration of PGE2, PGE1, and PGA2 inhibited proliferation. The potency ranking for PG-mediated inhibition of cell proliferation was identical to that found for PG-stimulated cAMP production. We conclude that all the naturally occurring PGs tested can activate the two signal transduction systems in osteoblasts.(ABSTRACT TRUNCATED AT 250 WORDS)
Using a recently described automated system, we measured total respiratory resistance (Rrs) of 59 patients with obstructive lung disease during spontaneous tidal breathing. We also measured airway resistance (Raw) and various other indices of pulmonary function. The results demonstrate that this system gives values of Rrs which correlate well with other objective measures of the severity of obstructive lung disease. It can also be used to detect changes in resistance after inhalation of a bronchodilator. In 10 patients we measured Raw during tidal breathing and computed the total non-airway resistance (Rnaw) as the difference between Rrs and Raw. Rnaw made up a larger portion of Rrs than would be expected in these patients. These results could not be accounted for by differences in breathing frequency, and may instead have been caused by differences between the times in the ventilatory cycle at which Rrs and Raw were measured. We cannot exclude the possibility that Rnaw was abnormally elevated in these patients.