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Biomedical subjects

N King

Publications and source records attributed to N King.

At least 19 recordsLinked to original sources
PubMed2002

The effect of graded levels of exercise on energy intake and balance in free-living men, consuming their normal diet.

OBJECTIVE: To assess the effect of graded increases in exercised-induced energy expenditure (EE) on appetite, energy intake (EI), total daily EE and body weight in men living in their normal environment and consuming their usual diets. DESIGN: Within-subject, repeated measures design. Six men (mean (s.d.) age 31.0 (5.0) y; weight 75.1 (15.96) kg; height 1.79 (0.10) m; body mass index (BMI) 23.3(2.4) kg/m(2)), were each studied three times during a 9 day protocol, corresponding to prescriptions of no exercise, (control) (Nex; 0 MJ/day), medium exercise level (Mex; approximately 1.6 MJ/day) and high exercise level (Hex; approximately 3.2 MJ/day). On days 1-2 subjects were given a medium fat (MF) maintenance diet (1.6 x resting metabolic rate (RMR)). MEASUREMENTS: On days 3-9 subjects self-recorded dietary intake using a food diary and self-weighed intake. EE was assessed by continual heart rate monitoring, using the modified FLEX method. Subjects' HR (heart rate) was individually calibrated against submaximal VO(2) during incremental exercise tests at the beginning and end of each 9 day study period. Respiratory exchange was measured by indirect calorimetry. Subjects completed hourly hunger ratings during waking hours to record subjective sensations of hunger and appetite. Body weight was measured daily. RESULTS: EE amounted to 11.7, 12.9 and 16.8 MJ/day (F(2,10)=48.26; P<0.001 (s.e.d=0.55)) on the Nex, Mex and Hex treatments, respectively. The corresponding values for EI were 11.6, 11.8 and 11.8 MJ/day (F(2,10)=0.10; P=0.910 (s.e.d.=0.10)), respectively. There were no treatment effects on hunger, appetite or body weight, but there was evidence of weight loss on the Hex treatment. CONCLUSION: Increasing EE did not lead to compensation of EI over 7 days. However, total daily EE tended to decrease over time on the two exercise treatments. Lean men appear able to tolerate a considerable negative energy balance, induced by exercise, over 7 days without invoking compensatory increases in EI.

Adult↗
PubMed2002

The effect of graded levels of exercise on energy intake and balance in free-living women.

AIM: We assessed the effect of graded increases in exercised-induced energy expenditure (EE) on appetite, daily energy intake (EI), total daily EE and body weight in six lean women using a within-subject, repeated measures design. METHOD: Subjects were each studied three times during 7 day treatments, corresponding to no-exercise (control; Nex; 0 MJ/day), medium exercise level (Mex; approximately 1.9 MJ/day) and high exercise level (Hex; approximately 3.4 MJ/day), with 2 day maintenance beforehand. Subjects self-weighed ad libitum food intake. EE was assessed by continual heart rate monitoring. During waking hours subjects recorded hourly sensations of hunger and appetite. RESULTS: EE amounted to 9.2, 11.0 and 12.1 MJ/day (F (2, 10)=5.67; P=0.023 (s.e.d.=0.87)) on the Nex, Mex and Hex treatments, respectively. The corresponding values for EI were 8.9, 9.2 and 10.0 MJ/day (F (2, 10)=4.80; P=0.035 (s.e.d.=0.36)). There were very weak treatment effects on hunger. Weight loss was significantly different from zero on the Mex and Hex treatments. CONCLUSION: Markedly increasing EE through exercise produced significant but partial compensations in EI ( approximately 33% of EE due to exercise). Accurate adjustments of El to acute increases in EE are likely to take weeks rather than days.

Adult↗
PubMed2001

A receptor tyrosine kinase from choanoflagellates: molecular insights into early animal evolution.

The evolution of the Metazoa from protozoans is one of the major milestones in life's history. The genetic and developmental events involved in this evolutionary transition are unknown but may have involved the evolution of genes required for signaling and gene regulation in metazoans. The genome of animal ancestors may be reconstructed by identification of animal genes that are shared with related eukaryotes, particularly those that share a more recent ancestry and cell biology with animals. The choanoflagellates have long been suspected to be closer relatives of animals than are fungi, the closest outgroup of animals for which comparative genomic information is available. Phylogenetic analyses of choanoflagellate and animal relationships based on small subunit rDNA sequence, however, have yielded ambiguous and conflicting results. We find that analyses of four conserved proteins from a unicellular choanoflagellate, Monosiga brevicollis, provide robust support for a close relationship between choanoflagellates and Metazoa, suggesting that comparison of the complement of expressed genes from choanoflagellates and animals may be informative concerning the early evolution of metazoan genomes. We have discovered in M. brevicollis the first receptor tyrosine kinase (RTK), to our knowledge, identified outside of the Metazoa, MBRTK1. The architecture of MBRTK1, which includes multiple extracellular ligand-binding domains, resembles that of RTKs in sponges and humans and suggests the ability to receive and transduce signals. Thus, choanoflagellates express genes involved in animal development that are not found in other eukaryotes and that may be linked to the origin of the Metazoa.

Amino Acid Sequence↗
PubMed2001

Haplotype study of three polymorphisms at the dopamine transporter locus confirm linkage to attention-deficit/hyperactivity disorder.

BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) is often treated using methylphenidate, a psychostimulant that inhibits the dopamine transporter. This led E.H. Cook and colleagues to consider the dopamine transporter locus (DAT1) as a primary candidate gene for ADHD. That group reported a significant association between ADHD and the 480-base pair (bp) allele of the variable number of tandem repeats (VNTR) polymorphism located in the 3' untranslated region of the DAT1 gene. This association was later replicated in additional studies. METHODS: The DAT1 gene has additional common polymorphisms in intron 9 and exon 9. We investigated the possibility of linkage of DAT1 and ADHD using the VNTR polymorphism and two additional common polymorphisms in 102 nuclear families with an ADHD proband. Using the transmission disequilibrium test, we examined the transmission of the alleles of each of these polymorphisms, as well as the haplotypes of the polymorphisms. RESULTS: We did not observe significant evidence for the biased transmission of the alleles of either the VNTR or the additional two polymorphisms when examined individually, although there was a trend for the biased transmission of the 480-bp allele of the VNTR. When we examined the haplotypes of the three polymorphisms we found significant evidence for biased transmission of one of the haplotypes containing the 480-bp VNTR allele. We also genotyped six additional DNA sequence variants of the DAT1 gene. However, these variants were not sufficiently polymorphic in our sample to be informative. Two of the DNA variants that result in an amino acid change, Ala559Val and Glu602Gly, were not observed in our sample. CONCLUSIONS: Our results support previous findings of an association between the DAT1 gene and ADHD.

Alleles↗
PubMed2001

Glutamate-loading stimulates metabolic flux and improves cell recovery following chemical hypoxia in isolated cardiomyocytes.

The amino acid glutamate is used in cardioplegic solutions, yet evidence is conflicting as to whether or not exogenous glutamate is indeed cardioprotective. This controversy may be because increasing extracellular glutamate does not necessarily lead to an increase in intracellular glutamate. In this study we aimed to determine whether isolation of myocytes in the presence of glutamate resulted in glutamate-loading of the cells, and, if so, whether such loading protected myocytes from simulated (chemical) hypoxia. Single ventricular myocytes were isolated from rat hearts in the presence and absence of 6.4 mM glutamate. Levels of glutamate and ATP were determined using HPLC, and NADH/NAD(+) was determined from cell autofluorescence. Chemical hypoxia was induced by superfusion with a solution containing 2.5 mM cyanide and no glucose. Intracellular [Ca(2+)] was measured by loading cells with indo-1, and cell length was measured using an edge-tracking device. Isolation of myocytes in the presence of glutamate resulted in increased intracellular glutamate levels compared with cells isolated in the absence of glutamate, 1324+/-108 v 948+/-124 pmol/mg protein, respectively (P<0.05). Cells loaded with glutamate showed increased NADH/NAD(+), (0.384+/-0.032 v 0.281+/-0.029, P<0.05) and greater ATP levels (36.031+/-1.633 nmol/mg protein v 19.279+/-3.327 nmol/mg protein, P<0.005) compared to control cells. When subjected to chemical hypoxia, cells underwent rigor-contracture at various timepoints, and were then reperfused following 5 min in rigor. Cells loaded with glutamate showed better recovery of diastolic [Ca(2+)], Ca(2+) transient amplitude, and improved contractile function compared with cells isolated in absence of glutamate. This study demonstrates an efficient method for loading myocytes with glutamate during cell isolation, and myocytes loaded with glutamate showed increased metabolic flux, as indexed by a higher NADH/NAD(+) and ATP content. Myocytes also exhibited better recovery from chemical hypoxia in terms of both Ca(2+) handling and cell contraction.

Adenosine Triphosphate↗
PubMed2001

Characteristics of L-aspartate transport and expression of EAAC-1 in sarcolemmal vesicles and isolated cells from rat heart.

OBJECTIVE: L-Aspartate is an important intermediary metabolite in the heart and has also been implicated in myocardial protection, but little is known about its transport across the cardiac sarcolemma. In this study we have tested the hypothesis that the high affinity sodium-dependent aspartate transporter, EAAC-1 is expressed in heart and have also characterised aspartate transport into the myocardium. METHODS: Characteristics of L-[14C]aspartate uptake into rat heart were investigated using sarcolemmal vesicles and isolated myocytes. The expression of EAAC-1 in the two preparations was also investigated by western blotting. RESULTS: The K(m) and V(max) of L-aspartate uptake was 9.78+/-0.7 microM and 1.17+/-0.27 pmol/mg/s in vesicles compared to 6.53+/-1.24 microM and 13.65+/-1.0 pmol/microl/s in cells. In vesicles, L-aspartate uptake was dependent on external sodium and internal potassium, and was rheogenic. In cells, L-aspartate uptake was also dependent on external sodium. Addition of unlabelled L- and D-aspartate and L-glutamate significantly inhibited L-[14C]aspartate uptake in both preparations but D-glutamate had no effect. An antibody to the aspartate transporter, EAAC-1 recognised a protein of appropriate size in both vesicles and cells. CONCLUSIONS: L-aspartate uptake in heart is mediated by a high affinity sodium-dependent transporter. This is accompanied by the expression in heart of EAAC-1. The physiological significance of this transporter with respect to aspartate utilisation in the heart is discussed.

Amino Acid Transport System X-AG↗
PubMed2001

Description and evaluation of a Newton-based electronic appetite rating system for temporal tracking of appetite in human subjects.

This study assessed the reliability and validity of a palm-top-based electronic appetite rating system (EARS) in relation to the traditional paper and pen method. Twenty healthy subjects [10 male (M) and 10 female (F)] - mean age M=31 years (S.D.=8), F=27 years (S.D.=5); mean BMI M=24 (S.D.=2), F=21 (S.D.=5) - participated in a 4-day protocol. Measurements were made on days 1 and 4. Subjects were given paper and an EARS to log hourly subjective motivation to eat during waking hours. Food intake and meal times were fixed. Subjects were given a maintenance diet (comprising 40% fat, 47% carbohydrate and 13% protein by energy) calculated at 1.6xResting Metabolic Rate (RMR), as three isoenergetic meals. Bland and Altman's test for bias between two measurement techniques found significant differences between EARS and paper and pen for two of eight responses (hunger and fullness). Regression analysis confirmed that there were no day, sex or order effects between ratings obtained using either technique. For 15 subjects, there was no significant difference between results, with a linear relationship between the two methods that explained most of the variance (r(2) ranged from 62.6 to 98.6). The slope for all subjects was less than 1, which was partly explained by a tendency for bias at the extreme end of results on the EARS technique. These data suggest that the EARS is a useful and reliable technique for real-time data collection in appetite research but that it should not be used interchangeably with paper and pen techniques.

Adult↗
PubMed2001

L-leucine transport in rat heart under normal conditions and effects of a simulated hypoxia.

L-leucine plays a central role in the regulation of protein metabolism in heart and has been implicated in myocardial protection, but little is known about the relationship between these phenomena and leucine transport across the cardiac sarcolemma. In this study we used sarcolemmal vesicles and ventricular myocytes isolated from rat heart to characterise L-leucine transport under normal conditions and to investigate the effect of simulated hypoxia or inhibition of protein synthesis. The Km and Vmax of leucine uptake were 5.24+/-0.65 mM and 1.43+/-1.84 nmol min(-1) mg(-1) protein in vesicles compared to 2.17+/-0.13 mM and 1.7+/-0.76 nmol min(-1) microl(-1) intracellular space in cells. Transport was not dependent on Na+ or H+ gradients. In vesicles L-leucine uptake was increased by trans-stimulation, whilst inhibition was observed with classical system L substrates including 2-aminobicyclo[2,2,1]-heptane-2-carboxylic acid (BCH) suggesting that this system mediated L-leucine transport in heart. L-Leucine uptake into isolated cardiac myocytes was inhibited after 20, 30 and 60 min of simulated hypoxia. This was not caused by reduced cell viability, although the cells underwent a rigor contracture. Inhibition of protein synthesis did not affect L-leucine transport.

Animals↗
PubMed2001

Idiopathic environmental intolerance: increased prevalence of panic disorder-associated cholecystokinin B receptor allele 7.

BACKGROUND: A growing body of evidence suggests that idiopathic environmental intolerance (IEI) is a psychophysiologic disorder with prominent features of anxiety/panic and somatization, although proponents of a toxicogenic explanation claim, despite a lack of convincing evidence, that symptoms arise from exposure to otherwise nonnoxious environmental agents. Patient behaviour is characterized by strenuous avoidance of perceived triggers to the point of severe impairment of normal social and vocational functioning. IEI proponents claim that previous studies showing a high prevalence of psychopathology in patients with IEI and studies showing panic responses to known panicogenic challenges merely reflect the anxiety-producing result of living with IEI. OBJECTIVE: We explored whether IEI and panic disorder, personality traits, or both shared an underlying neurogenetic basis that would predate the anxiety of IEI symptomatology. The DNA of patients with IEI was examined for the presence of known panic disorder-associated cholecystokinin B (CCK-B) receptor alleles and for personality trait-associated dopamine D4 receptor polymorphisms. METHODS: Eleven patients with typical IEI symptoms were recruited and were individually matched to normal control subjects from an existing bank for age, sex, and ethnic background. Genomic DNA was extracted from peripheral blood samples. CCK-B and dopamine D4 receptor polymorphisms were examined by using standard PCR-based techniques. RESULTS: There was a significantly higher prevalence of the panic disorder-associated CCK-B receptor allele 7 in subjects with IEI (9/22 [40.9%]) compared with control subjects (2/22 [9.1%], P =.037). There was no difference in personality trait-associated polymorphisms of the gene encoding dopamine D4 receptor between patients and control subjects. CONCLUSIONS: These findings provide preliminary evidence that IEI and panic disorder share a common neurogenetic basis, which would predate the anxiety-producing effects of IEI symptoms. Further studies with larger samples are warranted, but these results support previous studies that suggest that panic disorder may account for much of the symptomatology in at least some cases of IEI and provide a basis for rational treatment strategies.

Alleles↗
PubMed2001

Dopamine system genes not linked to social phobia.

Social phobia, particularly in its generalized form, has a genetic component in its etiology as suggested by positive twin studies and child temperament studies of social anxiety. Observations from functional imaging research suggest that dopamine function may be abnormal in the brains of patients with social phobia. Our investigation examined polymorphisms in the dopamine D2, D3 and D4 receptor genes, plus the dopamine transporter gene in a sample consisting of 17 multiplex social phobia families. We employed both parametric and non-parametric methods to test for linkage. Linkage was excluded for all loci under the broad diagnostic category. In the medium diagnostic category, the D3 receptor gene showed non-significant positive LOD scores (LOD = 0.62). We are able to clearly exclude a major effect for each of the four dopamine gene markers under the broad diagnosis of social phobia. Additional studies of dopamine system genes will be necessary to define clearly their role in social phobia.

Chromosome Mapping↗
PubMed2000

Using fMRI to study recovery from acquired dysphasia.

We have used functional magnetic resonance imaging (fMRI) to characterize brain activations associated with two distinct language tasks performed by a 28-year-old woman after partial recovery from dysphasia due to a left frontal hemispheric ischemic stroke. MRI showed that her ischemic lesion extended posteriorly from the left inferior frontal to the perisylvian cortex. fMRI scans of both language tasks revealed substantial differences in activation pattern relative to controls. The nature of this difference was task-specific. During performance of a verbal semantic decision task, the patient, in contrast to controls, activated a network of brain areas that excluded the inferior frontal gyrus (in either hemisphere). A second task involving rhyme judgment was designed to place a heavier cognitive load on language production processes and activated the left inferior frontal gyrus (Broca's area) strongly in normal controls. During this task, the most prominent frontal activation in the patient occurred in the right homologue of Broca's area. Subsequent analysis of this data by methods able to deal with responses of changing amplitude revealed additional, less sustained recruitment by the patient of cortex adjacent to the infarct in the region inferior to Broca's area during rhyming. These results suggest that in addition to changes in cognitive strategy, recovery from dysphasia could be mediated by both the preservation of neuronal networks in and around the infarct and the use of homologous regions in the contralateral hemisphere.

Adult↗
PubMed2000

The effect of body dissatisfaction on women's perceptions of female celebrities.

OBJECTIVE: Research suggests that media exposure causes some women to feel heightened dissatisfaction with their body shape. This study attempts to determine which women are effected as such, by investigating how women feel about their own bodies and how this effects their perceptions of female celebrities in the media. METHOD: Undergraduate females (n = 116) were shown one accurate and six distorted photographs of thin and heavy female celebrities. Each distorted photograph made the celebrity appear thinner or heavier than actuality. Participants chose which photograph portrayed each celebrity's true body shape. Body shape concerns were measured by the Body Shape Questionnaire. RESULTS: Women concerned about their body shape judged thin celebrities as thinner than actuality, whereas unconcerned women judged them accurately. Both groups judged heavy celebrities as heavier than actuality. DISCUSSION: Results suggest certain women are effected by media exposure due to their own perception of females in the media. Prevention strategies, and the media's role in body dissatisfaction and dieting disorders, are discussed.

Adolescent↗
PubMed2000

Apoptosis of vascular smooth muscle cells induced by cholesterol and its oxides in vitro and in vivo.

The ability of cholesterol and its oxides to induce apoptosis in vascular smooth muscle cells in tissue culture and in a rabbit model of atherosclerosis was evaluated. Apoptosis was detected using DNA laddering and in situ end-labelling of fragmented DNA. Cholesterol oxides, but not cholesterol, were found to inhibit proliferation and induce apoptosis of vascular smooth muscle cells in tissue culture. 7-ketocholesterol was found to be the most potent inhibitor of proliferation, while 25-hydroxycholesterol was found to be the most potent inducer of apoptosis. These data suggest that the inhibition of proliferation and the induction of apoptosis by cholesterol oxides within vascular smooth muscle cells use different pathways, suggesting a differential role for these cholesterol oxides within the arterial wall. Cholesterol feeding after balloon injury in a rabbit model of atherosclerosis is known to result in the accumulation of cholesterol oxides. However, we found that cholesterol feeding had no effect on the level of apoptosis in the rabbit aortic wall after balloon injury, suggesting that the major factor determining apoptosis in our model was the balloon injury.

Animals↗
PubMed2000

Research on the cognitive-behavioral treatment of school refusal: a review and recommendations.

Cognitive-behavior therapy is frequently used in the treatment of school refusal, a challenging problem for mental health professionals and school authorities. We review the clinical and research support for the efficacy of cognitive-behavior therapy using recently published guidelines for determining the level of evidentiary support for psychosocial interventions. Although cognitive-behavior therapy appears to be a useful treatment for school refusal, further research is needed before it can be considered as having "well-established" empirical status. Several other important methodological and theoretical issues are emphasized.

Adolescent↗
PubMed2000

The use of visual analogue scales to assess motivation to eat in human subjects: a review of their reliability and validity with an evaluation of new hand-held computerized systems for temporal tracking of appetite ratings.

This present paper reviews the reliability and validity of visual analogue scales (VAS) in terms of (1) their ability to predict feeding behaviour, (2) their sensitivity to experimental manipulations, and (3) their reproducibility. VAS correlate with, but do not reliably predict, energy intake to the extent that they could be used as a proxy of energy intake. They do predict meal initiation in subjects eating their normal diets in their normal environment. Under laboratory conditions, subjectively rated motivation to eat using VAS is sensitive to experimental manipulations and has been found to be reproducible in relation to those experimental regimens. Other work has found them not to be reproducible in relation to repeated protocols. On balance, it would appear, in as much as it is possible to quantify, that VAS exhibit a good degree of within-subject reliability and validity in that they predict with reasonable certainty, meal initiation and amount eaten, and are sensitive to experimental manipulations. This reliability and validity appears more pronounced under the controlled (but more artificial) conditions of the laboratory where the signal:noise ratio in experiments appears to be elevated relative to real life. It appears that VAS are best used in within-subject, repeated-measures designs where the effect of different treatments can be compared under similar circumstances. They are best used in conjunction with other measures (e.g. feeding behaviour, changes in plasma metabolites) rather than as proxies for these variables. New hand-held electronic appetite rating systems (EARS) have been developed to increase reliability of data capture and decrease investigator workload. Recent studies have compared these with traditional pen and paper (P&P) VAS. The EARS have been found to be sensitive to experimental manipulations and reproducible relative to P&P. However, subjects appear to exhibit a significantly more constrained use of the scale when using the EARS relative to the P&P. For this reason it is recommended that the two techniques are not used interchangeably.

Appetite↗
PubMed1999

Septation, dephosphorylation, and the activation of sigmaF during sporulation in Bacillus subtilis.

Cell-specific activation of transcription factor sigmaF during sporulation in Bacillus subtilis requires the formation of the polar septum and the activity of a serine phosphatase (SpoIIE) located in the septum. The SpoIIE phosphatase indirectly activates sigmaF by dephosphorylating a protein (SpoIIAA-P) in the pathway that controls the activity of the transcription factor. By use of a SpoIIE-GFP fusion protein in time-course and time-lapse experiments and by direct visualization of septa in living cells, we show that SpoIIE is present in the predivisional sporangium, where it often localizes near both cell poles in structures known as E-rings. We also present evidence consistent with the view that SpoIIE is present in both progeny cells after polar division. These findings are incompatible with a model for the control of sigmaF activity in which the phosphatase is simply sequestered to one cell. Instead, we conclude that the function of SpoIIE is subject to regulation, and we present evidence that this occurs in two stages. The first stage, which involves the phosphatase function of SpoIIE, depends on the cell division protein FtsZ and could correspond to the FtsZ-dependent assembly of SpoIIE into E-rings. The second stage occurs after the dephosphorylation of SpoIIAA-P and is dependent on the later-acting, cell-division protein DivIC. Evidence based on the use of modified and mutant forms of the phosphatase protein indicates that SpoIIE blocks the capacity of unphosphorylated SpoIIAA to activate sigmaF until formation of the polar septum is completed.

Bacillus subtilis↗