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G A Smith

Publications and source records attributed to G A Smith.

At least 163 records · Page 9Linked to original sources
PubMed1988

Design and properties of a fluorescent indicator of intracellular free Na+ concentration.

We have recently described a cryptand structure, FCryp-1, with appropriate properties for an indicator of intracellular free Na+ concentration using the 19F-n.m.r. chemical shift of the incorporated 5FBAPTA [1,2-bis-(2-amino-5-fluorophenoxy)ethane-NNN'N'-tetra-acetic acid] reporter group to measure the free cytosolic Na+ concentration [( Na+]i) [Smith, Morris, Hesketh and Metcalfe (1986) Biochim. Biophys. Acta 889, 82-83]. FCryp-1 carries four carboxylate groups to confer aqueous solubility and the indicator is membrane-permeant when the carboxyls are esterified with acetoxymethyl ester groups. Here we describe the synthesis of FCryp-2 to provide a fluorescent indicator of [Na+]i. FCryp-2 retains the parent tribenzo (2:2:1) cryptand structure of FCryp-1, in which the benzenoid ring at C-21 in FCryp-1 is replaced by an indole derivative which acts as the fluorophor in FCryp-2. With excitation at 340 nm, FCryp-2 gives an emission maximum at 460 nm in the absence of Na+ which shifts to 395 nm when FCryp-2 is saturated with Na+, with an isosbestic point at 455 nm. The apparent dissociation constant of FCryp-2 in a buffer solution of 100 mM-KCl/20 mM-KH2PO4/K2HPO4, pH 7.0, at 37 degrees C is 6.0 mM and the free Na+ concentration can be measured either from the calibrated fluorescence intensity at 395 nm, which increases 25-fold when Na+ is bound to FCryp-2, or from the ratio of fluorescence intensities at 395 nm and 455 nm. The measurement of free [Na+] by either method is unaffected by K+, Ca2+ or Mg2+ in the normal intracellular concentration ranges. Free [Na+] measurements by the ratio method are unaffected by pH from 6.6 to 7.6.

Ethers, Cyclic↗
PubMed1988

Orthodontic bonding to porcelain--bond strength and refinishing.

Glazed porcelain surfaces are not amenable to resin penetration for orthodontic bonding. The aims of this study were to evaluate (1) the bond strengths of two orthodontic composite materials to treated porcelain surfaces, (2) the effect of thermocycling, and (3) the porcelain surfaces after refinishing. Sixty glazed porcelain disks were assigned to three surface treatment groups: Silane (S), roughening and silane (RS), and roughening (R). The acid-etched enamel surfaces of 20 extracted teeth served as a control group. Ten specimens in each group were bonded with either Concise or System 1. They were stored in water at 37 degrees C or thermocycled 8 degrees to 45 degrees C. Shear bond strength was tested with an Instron testing machine (0.05 cm/min). Analysis of variance showed that the four surface treatment groups and the two composite materials produced significantly different bond strengths (P less than 0.01 and P less than 0.05, respectively). The effect of thermocycling was not significant (P greater than 0.05). Acid-etched enamel/Concise produced the highest mean bond strength (17.4 MN/m2). This was significantly greater (P less than 0.05) than the mean bond strengths of S/Concise (11.1), RS/System 1 (8.6), RS/Concise (8.1), and enamel/System 1 (7.8). The latter four mean bond strengths in turn were significantly greater (P less than 0.05) than S/System 1 (2.5), R/Concise (2.1), and R/System 1 (1.8). Of the four refinishing systems evaluated, all produced smooth surfaces but were unable to reproduce a glazed appearance. Roughening of porcelain and the silane treatment achieve bond strengths that should be clinically successful.(ABSTRACT TRUNCATED AT 250 WORDS)

Adhesives↗
PubMed1988

Age, variability, and speed: between-subjects diversity.

Two independent data sets were selected to examine the interrelations among reaction time (RT), between-subject variability or diversity (SD), and age. In both data sets, a strong correlation between RT and SD was obtained. This strong correlation was not affected when age was controlled in a partial correlation analysis. On the other hand, a weaker but significant correlation was obtained between age and SD. This correlation was eliminated when RT was controlled in a partial correlation analysis. Our analyses of the two data sets also indicated that the relation between RT and SD is identical for both young and elderly groups. Thus, the greater diversity often observed in performances of older groups is a direct consequence of slowing, rather than an independent effect of advancing age.

Adolescent↗
PubMed1988

Comparison between the cellobiose/mannitol and 51Cr-labelled ethylenediaminetetra-acetate absorption tests in the detection of coeliac disease.

1. The cellobiose/mannitol and 51Cr-labelled ethylenediaminetetra-acetate (51Cr-EDTA) absorption tests have been compared with respect to their ability to detect altered intestinal permeability in coeliac disease. 2. In patients with clinically proven coeliac disease, 13 out of 14 (93%) had abnormal urinary cellobiose/mannitol ratios, while only five out of 12 (42%) had abnormal urinary recoveries of 51Cr-EDTA. Thus, the cellobiose/mannitol absorption test is more sensitive in recognizing coeliac disease and this difference was significant (P less than 0.02). 3. Possible reasons for this difference in the sensitivity of the two absorption tests are discussed.

Adult↗
PubMed1988

Ca2+ transient, Mg2+, and pH measurements in the cardiac cycle by 19F NMR.

19F NMR indicators have been used to measure the free cytosolic cation concentrations ([Mn+]i, where M is the atomic symbol and n is the value of the charge) of Ca2+, H+, and Mg2+ in perfused ferret hearts. The [Ca2+]i transient, cytosolic pH (pHi), and [Mg2+]i have also been followed at 16 phases in the cardiac cycle in hearts paced at 1.25 Hz at 30 degrees C. The initial [Ca2+]i rose rapidly after a 50-ms delay, was maximal at greater than 1.5 microM after 150 ms, and declined thereafter to the initial concentration. In contrast, no significant changes in pHi (pH 7.03 +/- 0.08) or [Mg2+]i (1.2 +/- 0.1 mM) were detected in the cycle. A decrease in developed pressure when the [Ca2+]i indicator (but not the pHi or [Mg2+]i indicator) was loaded into hearts was substantially reversed by the addition of 50 microM ZnCl2 to the perfusion medium. The Zn2+ was taken up into the myoplasm and displaced Ca2+ bound to the indicator, a symmetrically substituted difluoro derivative of 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA), as evidenced by the appearance of the Zn-5FBAPTA resonance. The decrease in developed pressure caused by 5FBAPTA, therefore, may be due to its Ca2+ buffering effect on the myoplasm. By coloading hearts with the [Ca2+]i and pHi indicators, simultaneous measurement of several [Mn+]i was demonstrated, which should provide a useful addition to the methods available to monitor cardiac function and pharmacology.

Animals↗
PubMed1988

Caffeine reduction as an adjunct to anxiety management.

A reduction in tea and coffee drinking is popularly advocated for the relief of tension and anxiety symptoms, and it was decided to collect empirical data on this matter in the course of normal clinical practice. It was found that patients who made substantial reductions in caffeine drinking also made the greatest improvements in anxiety, irritability, sleep disturbance, headaches and abdominal symptoms. The methodology leaves an ambiguity in the interpretation of this result, but the data provide a useful basis for the design of further studies.

Adolescent↗
PubMed1988

[Energy and protein requirements of dogs and cats].

The latest research findings published on the energy and amino acid requirements of dogs and cats are reviewed. Special attention was paid to research results, applicable to working dogs, which have not as yet been incorporated into the recommended requirement standards of the National Research Council.

Animals↗
PubMed1987

Synergistic signals in the mechanism of antigen-induced exocytosis in 2H3 cells: evidence for an unidentified signal required for histamine release.

The aim of this study was to determine whether the increase in cytosolic free Ca2+ concentration ([Ca2+]i) in response to antigen (aggregated ovalbumin) on IgE-primed 2H3 cells was sufficient to account for exocytosis. When the [Ca2+]i responses to antigen and the Ca2+ ionophore A23187 were compared, A23187 was much less effective at releasing histamine at equivalent [Ca2+]i increases, and little or no stimulated histamine release occurred with A23187 concentrations that matched the [Ca2+]i response to antigen concentrations that stimulated maximal histamine release. The [Ca2+]i response to antigen is not, therefore, sufficient to account for exocytosis, although extracellular Ca2+ is necessary to initiate both the [Ca2+]i response and histamine release: the antigen must generate an additional, unidentified, signal that is required for exocytosis. To determine whether this signal was the activation of protein kinase C, the effects of the phorbol ester 12-0-tetradecanoyl phorbol 13-acetate (TPA) on the responses to antigen were examined. TPA blocked the antigen-induced [Ca2+]i response and the release of inositol phosphates but had little effect on histamine release and did not stimulate exocytosis by itself. The unidentified signal from the antigen is therefore distinct from the activation of protein kinase C and is generated independently of the [Ca2+]i response or the release of inositol phosphates. Taken together with other data that imply that there is very little activation of protein kinase C by antigen when the rate of histamine release is maximal, it is concluded that the normal exocytotic response to antigen requires the synergistic action of the [Ca2+]i signal together with an unidentified signal that is not mediated by protein kinase C.

Antigens↗
PubMed1987

Relationship of androgenic activity to splanchnic insulin metabolism and peripheral glucose utilization in premenopausal women.

The importance of androgenic activity in mediating the effects of obesity and body fat topography on splanchnic insulin metabolism and peripheral insulin sensitivity was studied in 19 nonhirsute premenopausal women with a wide range of ideal body weight [percent ideal body weight (% IBW), 78-202%] and body fat distribution pattern [waist to hip girth ratio (WHR), 0.67-0.91]. Turnover kinetics of peripheral plasma C-peptide and insulin were measured, and estimates of pancreatic insulin production (PIP) and the hepatic extraction fraction (HEF) were calculated. The peripheral insulin sensitivity index (M/I) was determined during an euglycemic insulin clamp study. Androgenic activity was assessed by estimating the plasma level of sex hormone-binding globulin (SHBG) and percentage of free testosterone (% FT). After iv glucose stimulation, PIP ranged from 40-254 mU/min X m2 and correlated highly with % IBW (r = 0.78; P less than 0.01). Insulin HEF ranged from 5-69% of the pancreatic production and was inversely proportional to WHR (r = -0.60; P less than 0.01). Increasing WHR also correlated with the diminution in M/I (r = -0.47; P less than 0.05), which, in turn, correlated with the decline in the HEF of insulin (r = 0.60; P less than 0.01). Since PIP, HEF, and M/I correlated with SHBG and % FT, and since the degree of androgenic activity correlated with % IBW and WHR, partial regression analysis was performed. After adjusting for the effects of SHBG and % FT, the relationship between % IBW and PIP remained unaltered, whereas the correlation between WHR and HEF or M/I and their relationship to each other were either markedly reduced or became insignificant. Thus, in premenopausal women, the increase in pancreatic insulin production with increasing weight is independent of the degree of androgenic activity. On the other hand, the decline in hepatic insulin extraction and diminution in peripheral insulin sensitivity with upper body fat localization are in part mediated by increased androgenic activity. This association may account for the pronounced hyperinsulinemia and insulin resistance characteristic of this form of obesity.

Adipose Tissue↗
PubMed1986

Design of an indicator of intracellular free Na+ concentration using 19F-NMR.

The development is described of an Na+ chelator with appropriate properties for an indicator of intracellular free Na+ concentration ([Na+]i). The new indicator, FCryp-1, is a tribenzo derivative of the parent (2:2:1) cryptand structure, incorporating the same F-substituted dibenzo 19F-NMR reporter group as the free [Ca2+] indicator, 5FBAPTA (Smith, G.A., Hesketh, T.R., Metcalfe, J.C., Feeney, J. and Morris, P.G. (1983) Proc. Natl. Acad. Sci., USA 80, 7178-7182). FCryp-1 has appropriate affinity for Na+ (KNa = 10(1.3) M-1) and selectivity over other intracellular cations (KK; KCa; K Mg less than 10(-1) M(-1)) for a [Na]i indicator. There is an 19F-NMR chemical shift of 2.00 ppm between free FCryp-1 and the Na-FCryp-1 complex which provides a direct read out of free [Na+]. FCryp-1 carries four carboxylate groups to confer aqueous solubility which can be esterified with acetoxymethyl groups to render the indicator membrane permeant. Experiments on pig lymphocytes loaded with FCryp-1 gave an indicated [Na+]i of 13.8 +/- 1.8 mM (n = 4). The FCryp-1 structure can also be readily modified to provide fluorescent [Na+]i indicators.

Animals↗
PubMed1986

Splanchnic insulin metabolism in obesity. Influence of body fat distribution.

The effects of obesity and body fat distribution on splanchnic insulin metabolism and the relationship to peripheral insulin sensitivity were assessed in 6 nonobese and 16 obese premenopausal women. When compared with the nonobese women, obese women had significantly greater prehepatic production and portal vein levels of insulin both basally and following glucose stimulation. This increase correlated with the degree of adiposity but not with waist-to-hip girth ratio (WHR). WHR, however, correlated inversely with the hepatic extraction fraction and directly with the posthepatic delivery of insulin. The latter correlated with the degree of peripheral insulinemia. The decline in hepatic insulin extraction with increasing WHR also correlated with the accompanying diminution in peripheral insulin sensitivity. Increasing adiposity is thus associated with insulin hypersecretion. The pronounced hyperinsulinemia of upper body fat localization, however, is due to an additional defect in hepatic insulin extraction. This defect is closely allied with the decline in peripheral insulin sensitivity.

Adipose Tissue↗