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J C Smith

Publications and source records attributed to J C Smith.

At least 145 records · Page 8Linked to original sources
PubMed1998

Characterization of beta-carotene 15,15'-dioxygenase activity in TC7 clone of human intestinal cell line Caco-2.

The purpose of this study was to identify mammalian cell line(s) which possess intrinsic enzymatic activity of beta-carotene 15, 15'-dioxygenase. This enzyme (EC1.13.11.21) converts beta-carotene to retinal (precursor of retinol and retinoic acid). To assess activity, cellular enzyme preparations were incubated with beta-carotene for 60 min; retinal formed was quantified by HPLC. Activity was not detected in IPEC-1, HepG2, HL60, Wurzburg, or parent Caco-2 cell lines. However, two subclones of Caco-2, PF11 and TC7, possessed activity (2.5 and 14.7 pmol/h.mg, respectively). Using the enzyme preparation of TC7 cells, retinal formation was linear with incubation time and protein concentration; Km and Vm values were 1.6 microM and 23.8 pmol/h.mg, respectively. In addition, when TC7 cells were maintained in serum-free medium, activity was increased 8.2-fold after 19 days of postconfluency. Finally, 48 h incubation with beta-carotene (delivered to TC7 cells in Tween 40) resulted in a 1.7-fold increase of dioxygenase activity and the appearance of vitamin A (9.3 pmol/mg protein). However, retinoic acid was not detected under our experimental conditions. In sum, the TC7 subclone of the Caco-2 cell line possesses beta-carotene 15, 15'-dioxygenase activity and thus can be useful in future investigations of human carotenoid metabolism.

Caco-2 Cells↗
PubMed1998

Abortion surveillance--United States, 1995.

CONDITION: Since 1990 (i.e., the year in which the number of abortions was highest), the annual number of abortions in the United States has decreased by 15%. REPORTING PERIOD COVERED: This report summarizes and reviews information reported to CDC regarding legal induced abortions obtained in the United States during 1995. DESCRIPTION OF SYSTEM: For each year since 1969, CDC has compiled abortion data received from 52 reporting areas: 50 states, the District of Columbia, and New York City. RESULTS: In 1995, a total of 1,210,883 legal abortions were reported to CDC, representing a 4.5% decrease from the number reported for 1994. The abortion ratio was 311 legal induced abortions per 1,000 live births, and the abortion rate was 20 per 1,000 women aged 15-44 years, the lowest ratio and rate recorded since 1975. Women who were undergoing an abortion were more likely to be young, white, and unmarried; most were obtaining an abortion for the first time. Approximately half of all abortions (54%) were performed at < or =8 weeks of gestation, and approximately 88% were performed before 13 weeks. Approximately 16% of abortions were performed at the earliest weeks of gestation (< or =6 weeks), approximately 17% at 7 weeks of gestation, and approximately 21% at 8 weeks of gestation. Few abortions were provided after 15 weeks of gestation -- approximately 4% of abortions were obtained at 16-20 weeks, and 1.4% were obtained at > or =21 weeks. Younger women (i.e., women aged < or =24 years) were more likely to obtain abortions later in pregnancy than were older women. INTERPRETATION: Since 1990, the number of abortions has declined each year. Since 1987, the abortion-to-live-birth ratio has declined; in 1995, it was the lowest recorded since 1975. This decrease in the abortion ratio reflects a trend that a lower proportion of pregnant women obtain induced abortion. ACTIONS TAKEN: The number and characteristics of women who obtain abortions in the United States should continue to be monitored so that trends in induced abortion can be assessed, efforts to prevent unintended pregnancy can be evaluated, and the preventable causes of morbidity and mortality associated with abortions can be identified and reduced.

Abortion, Legal↗
PubMed1998

Mammalian bombesin-like peptides extend the intermeal interval in freely feeding rats.

We evaluated the effects of systemic delivery of amphibian bombesin and its mammalian homologues on the length of the postprandial intermeal interval. Adult male rats, feeding ad libitum, were injected intraperitoneally (i.p.) 5 min after the end of the first nocturnal meal with 0 (vehicle), 2.5, 5, or 10 nM/kg of tetradecapeptide bombesin (BN), gastrin-releasing peptide(1-27) (GRP1-27), the C-terminal decapeptide of GRP(18-27) (GRP18-27), the C-terminal decapeptide of neuromedin B (NMB23-32), or combinations of equimolar doses of GRP1-27 and NMB23-32. BN produced a potent, dose-related extension (maximum of 177%) of the first postprandial intermeal interval; GRP1-27 produced a lesser but significant prolongation (maximum of 47%); the combination of GRP1-27 and NMB23-32 produced an intermediate prolongation (maximum of 70%); GRP18-27 alone and NMB23-32 alone failed to produce any significant change. Peptide effects were limited to the first postprandial intermeal interval. The results demonstrate that systemic, postprandial injection of BN, GRP1-27, or the combination of GRP1-27 and NMB23-32 extends the duration of the postprandial intermeal interval. The results suggest that the endogenous peptides, released in the gastrointestinal tract by ingested food, have a potent satiety action, selectively lengthening the intermeal interval.

Animals↗
PubMed1998

X-ray diffuse scattering and rigid-body motion in crystalline lysozyme probed by molecular dynamics simulation.

Rigid-body motions are determined from a 1 ns molecular dynamics simulation of the unit cell of orthorhombic hen egg-white lysozyme and their contribution to X-ray diffuse scattering intensities are examined. Using a dynamical cluster technique, groups of backbone atoms that move as approximately rigid bodies are derived from the intramolecular interatomic fluctuation matrix. These groups tend to be local in the sequence or connected by disulphide bonds, and contain on average five residues each, X-ray diffuse scattering patterns, which are sensitive to collective motions, are calculated from the full simulation trajectory (including all the protein degrees of freedom). The results reproduce the main features of the experimental scattering. Diffuse scattering is also calculated from fitted trajectories of the rigid bodies. The full simulation diffuse scattering and atomic displacements are found to be well reproduced by a model in which the backbone atoms form the rigid groups determined using the dynamical cluster technique and the individual side-chains behave as separate rigid bodies: the resulting R-factor with the full simulation scattering is 5%. Quantitatively poorer agreement is obtained from trajectories in which the secondary structural elements of the protein are considered rigid. Rigid whole-molecule and domain motions make only minor contributions to the protein atom displacements. Finally, correlations in the interatomic fluctuations are examined directly using a canonical method.

Computer Simulation↗
PubMed1998

The rat's acceptance and preference for sucrose, maltodextrin, and saccharin solutions and mixtures.

Prior work indicates that rats prefer a mixture of sucrose and maltodextrin to either carbohydrate alone. The present experiment examined whether a sucrose + maltodextrin (S + M) mixture also increases total fluid intake as does saccharin + carbohydrate mixtures. In 23 h/day, one-bottle tests male rats consumed more of a 1% sucrose + 1% maltodextrin mixture than they did of either 2% sucrose or 2% maltodextrin. Adding 0.2% saccharin to the S + M mixture further stimulated consumption. The rats overconsumed the mixture solutions primarily by increasing bout size. In two-bottle choice tests the rats strongly preferred the S + M mixture to the 2% sucrose and 2% maltodextrin solutions and showed a nonreliable preference for the S + M + saccharin mixture to the S + M mixture. The palatability of the S + M mixture is thought to be related to the activation of separate "sweet" and "maltodextrin" tastes.

Animals↗
PubMed1998

Magnetic field conditioned taste aversion in rats.

Conditioned taste aversion is a common classic conditioning procedure used to identify noxious stimuli. When a rat is given a taste solution, the conditioned stimulus (CS), followed by an unpleasant experience, the unconditioned stimulus (US), the rat will avoid consumption of the CS in future presentations. These experiments use the taste aversion procedure to examine the effect of exposure to a high magnetic field. A solution consisting of 3.0 g glucose and 1.25 g saccharin per 1 L of solution (G+S) was used as the CS and a 9.4-T magnet served as the US. In Experiment 1, all rats received a 10 min presentation of the G+S solution followed by either a 30 min exposure to the magnetic field (Magnet, n = 8), a 30-min exposure in a container with similar conditions but lacking the magnetic field (Sham, n = 8), or no exposure (Control, n = 8). The Magnet Group showed a taste aversion on the first day of preference testing (p < 0.05). Experiment 2 employed the same US-CS protocol for 3 consecutive days of conditioning. The Magnet Group demonstrated a taste aversion for the postexposure Days 1-8 (p < 0.01). There was no difference between the Sham and Control Groups in either experiment. The results of this study clearly demonstrate that the rats associated the G+S solution with the experience of being exposed to the high magnetic field and avoided the solution in subsequent presentations.

Animals↗
PubMed1998

Structural modeling of the complex between an acetylcholine receptor-mimicking antibody and its snake toxin antigen.

The antibody M alpha2-3 neutralizes the functional, acetylcholine receptor binding activity of its antigen, neurotoxin alpha, and exhibits several other properties in common with the receptor itself. We present here the results of calculations examining the three-dimensional structure of the toxin alpha:M alpha2-3 complex. The antigen structure, determined by nuclear magnetic resonance spectroscopy, was docked to models of the variable fragment of the antibody combining site by using a method based on surface complementarity and maximization of buried surface area while taking into account the possibility of conformational change on complexation. Extensive experimental information on the location of the functional epitope was incorporated into the analysis and used to screen candidate geometries of the complex resulting from the modeling. Eight plausible structures that are in accord with the experimental data were derived. Common structural features of the models are discussed, and residues of the antibody-combining site that are expected to play important roles in complexation are identified. In particular, three epitope residues that, according to mutagenesis experiments, make particularly strong contributions to the binding, interact excentrically and do not make contact with the central loops of the combining site, L3 and H3.

Animals↗
PubMed1998

Establishment of a BMP-4 morphogen gradient by long-range inhibition.

Recent work suggests that signaling molecules such as activin are capable of acting at long range to establish a morphogen gradient in the amphibian embryo and that responding cells activate different genes at distinct threshold levels of activin. Other signaling molecules like BMP-4 and Xnr-2 also exert concentration-dependent effects, but these factors appear to diffuse less freely. This raises the question of whether gradients of these inducing factors are indeed established, and if so, how they are generated. In this paper we demonstrate directly that BMP-4 elicits graded responses in gastrula-stage embryos. We then show that an effective BMP-4 gradient is established not by diffusion of BMP-4 protein but by the long-range effects of two BMP-4 inhibitors, noggin and chordin. This provides a novel mechanism for the establishment of a morphogen gradient in vertebrate embryos.

Animals↗
PubMed1998

Oxidation of tetrahydro-beta-carboline by cytochrome P-450cam--determination and rationalisation of product distribution.

Cytochrome P-450cam oxidises 1,2,3,4-tetrahydro-beta-carboline, an indolic alkaloid. We report here measurements of the product distribution of this oxidation. To rationalise the experimental results, ab initio quantum-chemistry calculations of the product stabilities and molecular-dynamics calculations of the substrate-binding mode in the active site were performed. The calculations suggest that the product distribution is influenced by both the relative intrinsic gas-phase stabilities of the monohydroxy products and by conformational rearrangement of the active site on substrate binding.

Binding Sites↗
PubMed1998

Excluded volume in the configurational distribution of a strongly-denatured protein.

The configurational distribution of phosphoglycerate kinase (PGK) strongly-denatured in 4 M guanidine hydrochloride solution is investigated using small-angle neutron scattering (SANS) and Monte Carlo computer simulation. It is shown that the experimental scattering profile can be represented by a random flexible chain of spheres of excess scattering density with excluded volume interactions, the best agreement being achieved when partial sphere intersection is allowed. The radius of gyration of the chain increases by a factor of 4 on denaturation, whereas the average length of segments approximately 5 residues long increases by only approximately 10%, consistent with a picture in which the large expansion on denaturation originates primarily from increased long-range flexibility of the polypeptide chain. The results provide a description of the chain statistics from which the construction of starting points for simulation studies of folding of the protein can be envisaged.

Computer Simulation↗
PubMed1998

Effect of oral creatine ingestion on parameters of the work rate-time relationship and time to exhaustion in high-intensity cycling.

The relationship between work rate (W) and time to exhaustion (t) during intense exercise is commonly described by either a hyperbolic function (NLin), t= W'/(W-Wcp), or by its linear equivalent (LinW) Wlim=W' + Wcp(t). The parameter Wcp (critical power) has been described as an inherent characteristic of the aerobic energy system, while W' has been shown to be a ralid estimate of anaerobic work capacity. Recent studies have demonstrated that oral supplementation of creatine monohydrate (CrH2O) increases total muscle creatine stores, and have linked these increases to improved performances in intense intermittent exercise. This study was conducted to determine the effect of CrH2O supplementation on estimates of W' and Wcp derived from the NLin and LinW equations, and to determine the effect of CrH2O on t in exhaustive constant power exercise of different intensities. Fifteen active but untrained university students completed three phases of testing on a cycle ergometer: (1) familiarization, three learning trials, (2) baseline determination of W' and Wcp, four bouts performed at a W selected to elicit fatigue in 90-600 s, and (3) experimental determination of W' and Wcp, four bouts performed at the same W as baseline, but performed after 5 days of ingesting either a placebo (4 x 6 g of glucose/day) or CrH2O (4 x 5 g of CrH2O and 1 g glucose/day). Testing was administered in a double-blind manner. Analyses of covariance revealed a significant effect for CrH2O on both estimates of W' (NLin, P=0.04; LinW, P < 0.01), but not on estimates of Wcp (NLin, P=0.37; LinW; P=0.30). Within groups, t was significantly different for only CrH2O at the two highest Ws (P=0.04). It is concluded that oral ingestion of CrH2O increases estimates of W' due to an improved t at the shorter, more intense exercise bouts.

Administration, Oral↗
PubMed1998

Enzyme activity below the dynamical transition at 220 K.

Enzyme activity requires the activation of anharmonic motions, such as jumps between potential energy wells. However, in general, the forms and time scales of the functionally important anharmonic dynamics coupled to motion along the reaction coordinate remain to be determined. In particular, the question arises whether the temperature-dependent dynamical transition from harmonic to anharmonic motion in proteins, which has been observed experimentally and using molecular dynamics simulation, involves the activation of motions required for enzyme function. Here we present parallel measurements of the activity and dynamics of a cryosolution of glutamate dehydrogenase as a function of temperature. The dynamical atomic fluctuations faster than approximately 100 ps were determined using neutron scattering. The results show that the enzyme remains active below the dynamical transition observed at approximately 220 K, i.e., at temperatures where no anharmonic motion is detected. Furthermore, the activity shows no significant deviation from Arrhenius behavior down to 190 K. The results indicate that the observed transition in the enzyme's dynamics is decoupled from the rate-limiting step along the reaction coordinate.

Catalysis↗
PubMed1998

SOX10 mutations in patients with Waardenburg-Hirschsprung disease.

Waardenburg syndrome (WS; deafness with pigmentary abnormalities) and Hirschsprung's disease (HSCR; aganglionic megacolon) are congenital disorders caused by defective function of the embryonic neural crest. WS and HSCR are associated in patients with Waardenburg-Shah syndrome (WS4), whose symptoms are reminiscent of the white coat-spotting and aganglionic megacolon displayed by the mouse mutants Dom (Dominant megacolon), piebald-lethal (sl) and lethal spotting (ls). The sl and ls phenotypes are caused by mutations in the genes encoding the Endothelin-B receptor (Ednrb) and Endothelin 3 (Edn3), respectively. The identification of Sox10 as the gene mutated in Dom mice (B.H. et al., manuscript submitted) prompted us to analyse the role of its human homologue SOX10 in neural crest defects. Here we show that patients from four families with WS4 have mutations in SOX10, whereas no mutation could be detected in patients with HSCR alone. These mutations are likely to result in haploinsufficiency of the SOX10 product. Our findings further define the locus heterogeneity of Waardenburg-Hirschsprung syndromes, and point to an essential role of SOX10 in the development of two neural crest-derived human cell lineages.

Amino Acid Sequence↗
PubMed1998

Modelling and simulation of light-activated membrane proteins: dynamical transitions in bacteriorhodopsin.

Many of the functions of membranes are carried out by proteins associated with them. A knowledge of atomic-detail membrane protein structures and dynamics is required for a full understanding of these functions. We briefly discuss recent progress in this field using modelling and simulation. One of the best characterised membrane proteins, bacteriorhodopsin, undergoes dynamical transitions with temperature. Here we present preliminary results of molecular dynamics simulation of this protein as a function of temperature, indicating the presence of dynamical transitions at approximately the temperatures seen experimentally.

Bacteriorhodopsins↗
PubMed1998

Structural modeling of the pro-ocytocin-neurophysin precursor.

The hormonal precursor pro-ocytocin-neurophysin is activated by selective cleavage at Arg2-Ala13, producing mature ocytocin and neurophysin. To understand the cleavage mechanism better, and in particular the recognition of the cleavage site, it is necessary to characterize the three-dimensional structure of the precursor molecule. Here we combine a variety of experimental data with molecular modeling and dynamics calculations to derive possible precursor conformations. In the models obtained, the N-terminus of the precursor, corresponding to the ocytocin segment, is hydrogen bonded in a pocket of the neurophysin moiety in a similar manner to a crystallographically obtained non-covalent complex between the two molecules. The calculations suggest that although the ocytocin segment is relatively flexible, it adopts a stable, broad loop structure in the vicinity of the cleavage region, which may constitute the structural element recognized by the cleaving enzyme. The calculations also suggest a possible widening of the distance between the two neurophysin domains in the precursor relative to that in the non-covalent neurophysin-ocytocin complex.

Amino Acid Sequence↗
PubMed1998

Management of substernal goiter.

OBJECTIVE: To analyze the presentation, evaluation and treatment of patients with large substernal goiters, with emphasis on the radiographic evaluation and the results of treatment. STUDY DESIGN: A retrospective chart review of 150 patients undergoing thyroidectomy at the Vanderbilt University Department of Otolaryngology-Head and Neck Surgery. METHODS: Charts of patients undergoing thyroidectomy were reviewed. Those with substernal goiter, defined as a major portion of the goiter within the mediastinum, were included in the study. When available, the radiographic studies were reviewed by a staff neuroradiologist. RESULTS: Twenty-three patients (15.3%) presented with substernal extension of the goiter. Characteristics of these patients included mean age of 59 years, 78% female, symptoms of compression such as dyspnea, choking, and dysphagia (65%), hoarseness (43%), and previous thyroid surgery (30%). Seventeen percent were asymptomatic. Preoperative radiographs demonstrated tracheal compression (73%), tracheal deviation (77%), esophageal compression (27%), and major vessel displacement (50%). Histology revealed multinodular goiter (16/23, 70%), thyroiditis (3/23, 13%), and malignancy (4/23, 17%). The average size of the resected specimen in greatest dimension was 8.0 cm (range, 3.0-14.0 cm) and weighed 148 g (range, 39-426 g). All were successfully approached through a transcervical incision without the need for sternotomy, and total thyroidectomy was performed in 83% of the cases. No major complications have been documented, and no evidence of tracheomalacia was encountered. CONCLUSION: Despite the large size of these goiters and the significant involvement of the major mediastinal structures, all were approached through the transcervical incision. Further, despite significant tracheal involvement, there were no cases of tracheomalacia or major complications. For intraoperative planning, the authors advocate the routine use of preoperative computed tomography scanning.

Adult↗
PubMed1998

The T-box transcription factor Brachyury regulates expression of eFGF through binding to a non-palindromic response element.

Brachyury is a member of the T-box gene family and is required for formation of posterior mesoderm and notochord during vertebrate development. The ability of Brachyury to activate transcription is essential for its biological function, but nothing is known about its target genes. Here we demonstrate that Xenopus Brachyury directly regulates expression of eFGF by binding to an element positioned approximately 1 kb upstream of the eFGF transcription start site. This site comprises half of the palindromic sequence previously identified by binding site selection and is also present in the promoters of the human and mouse homologues of eFGF.

Animals↗