Clustering of diet, physical activity and sedentary behaviour among Australian children: cross-sectional and longitudinal associations with overweight and obesity.

BACKGROUND/OBJECTIVES: Evidence suggests diet, physical activity (PA) and sedentary behaviour cluster together in children, but research supporting an association with overweight/obesity is equivocal. Furthermore, the stability of clusters over time is unknown. The aim of this study was to examine the clustering of diet, PA and sedentary behaviour in Australian children and cross-sectional and longitudinal associations with overweight/obesity. Stability of obesity-related clusters over 3 years was also examined. SUBJECTS/METHODS: Data were drawn from the baseline (T1: 2002/2003) and follow-up waves (T2: 2005/2006) of the Health Eating and Play Study. Parents of Australian children aged 5-6 (n=87) and 10-12 years (n=123) completed questionnaires. Children wore accelerometers and height and weight were measured. Obesity-related clusters were determined using K-medians cluster analysis. Multivariate regression models assessed cross-sectional and longitudinal associations between cluster membership, and body mass index (BMI) Z-score and weight status. Kappa statistics assessed cluster stability over time. RESULTS: Three clusters, labelled 'most healthy', 'energy-dense (ED) consumers who watch TV' and 'high sedentary behaviour/low moderate-to-vigorous PA' were identified at baseline and at follow-up. No cross-sectional associations were found between cluster membership, and BMI Z-score or weight status at baseline. Longitudinally, children in the 'ED consumers who watch TV' cluster had a higher odds of being overweight/obese at follow-up (odds ratio=2.8; 95% confidence interval: 1.1, 6.9; P<0.05). Tracking of cluster membership was fair to moderate in younger (K=0.24; P=0.0001) and older children (K=0.46; P<0.0001). CONCLUSIONS: This study identified an unhealthy cluster of TV viewing with ED food/drink consumption, which predicted overweight/obesity in a small longitudinal sample of Australian children. Cluster stability was fair to moderate over 3 years and is a novel finding. Prospective research in larger samples is needed to examine how obesity-related clusters track over time and influence the development of overweight and obesity.

Left dorsolateral prefrontal cortex supports context-dependent prioritisation of off-task thought.

When environments lack compelling goals, humans often let their minds wander to thoughts with greater personal relevance; however, we currently do not understand how this context-dependent prioritisation process operates. Dorsolateral prefrontal cortex (DLPFC) maintains goal representations in a context-dependent manner. Here, we show this region is involved in prioritising off-task thought in an analogous way. In a whole brain analysis we established that neural activity in DLPFC is high both when 'on-task' under demanding conditions and 'off-task' in a non-demanding task. Furthermore, individuals who increase off-task thought when external demands decrease, show lower correlation between neural signals linked to external tasks and lateral regions of the DMN within DLPFC, as well as less cortical grey matter in regions sensitive to these external task relevant signals. We conclude humans prioritise daydreaming when environmental demands decrease by aligning cognition with their personal goals using DLPFC.

The role of energy intake and energy misreporting in the associations between eating patterns and adiposity.

BACKGROUND/OBJECTIVES: Research examining associations between eating occasion (EO) frequency and adiposity is inconclusive; studies examining the impact of energy misreporting are rare. This study examined associations between eating patterns and adiposity, with adjustment for energy misreporting, in a nationally representative sample of Australian adults. SUBJECTS/METHODS: Dietary intake was assessed via two 24-h recalls collected during the 2011-12 National Nutrition and Physical Activity Survey (n=4050 adults, aged ⩾19 years). Frequencies of all EOs, meals and snacks were calculated. Height, weight and waist circumference (WC) were measured. Energy misreporting was assessed as the ratio of energy intake to predicted energy expenditure (EI:EE). Energy misreporters were identified by EI:EE ratios, <0.68 or >1.32. Multivariate regression models assessed associations between eating patterns and body mass index (BMI), WC, overweight/obesity (BMI ⩾25 kg m-2) and central overweight/obesity (WC ⩾94 cm in men and ⩾80 cm in women). RESULTS: After adjustment for covariates and EI:EE, frequency of all EOs, meals (women only) and snacks was positively associated with WC and BMI (all P<0.01). Snack, but not meal frequency, was also associated with overweight/obesity (men: OR=1.22, 95% CI 1.07-1.39; women: OR=1.26, 95% CI 1.10-1.43) and central overweight/obesity (men: OR=1.17, 95% CI 1.04-1.32; women: OR=1.21, 95% CI 1.06-1.37). Multivariate analysis that excluded energy misreporters and adjusted for EI yielded either null or inverse associations (P<0.05). CONCLUSIONS: These findings suggest that the associations between eating patterns and adiposity are complicated by the role of EI and energy misreporting. Longitudinal research that considers the impact of EI and energy misreporting is needed to better understand the relationship between eating patterns and obesity.

Significant Changes in Cell and Chloroplast Development in Young Wheat Leaves (Triticum aestivum cv Hereward) Grown in Elevated CO2.

Cell and chloroplast development were characterized in young Triticum aestivum cv Hereward leaves grown at ambient (350 [mu]L L-1) or at elevated (650 [mu]L L-1) CO2. In elevated CO2, cell and chloroplast expansion was accelerated by 10 and 25%, respectively, in the first leaf of 7-d-old wheat plants without disruption to the leaf developmental pattern. Elevated CO2 did not affect the number of chloroplasts in relation to mesophyll cell size or the linear relationship between chloroplast number or size and mesophyll cell size. No major changes in leaf anatomy or in chloroplast ultrastructure were detected as a result of growth in elevated CO2, but there was a marked reduction in starch accumulation. In leaf sections fluorescently tagged antisera were used to visualize and quantitate the amount of cytochrome f, the [alpha]- and [beta]-subunits of the coupling factor 1 in ATP synthase, D1 protein of the photosystem II reaction center, the 33-kD protein of the extrinsic oxygen-evolving complex, subunit II of photosystem I, and ribulose-1,5-bisphosphate carboxylase/oxygenase. A significant finding was that in 10 to 20% of the mesophyll cells grown in elevated CO2 the 33-kD protein of the extrinsic oxygen-evolving complex of photosystem II and cytochrome f were deficient by 75%, but the other proteins accumulated normally.

A Genetic Analysis of Chloroplast Division and Expansion in Arabidopsis thaliana.

A nuclear recessive mutant of Arabidopsis thaliana, arc5, has been isolated in which there is no significant increase in chloroplast number during leaf mesophyll cell expansion and in which there are only 13 chloroplasts per mesophyll cell compared with 121 in wild-type cells. Mature arc5 chloroplasts in fully expanded mesophyll cells are 6-fold larger than in wild-type cells. A large proportion of arc5 chloroplasts also show some degree of central constriction, suggesting that the mutation has prevented the completion of the chloroplast division process. To examine the interaction of arc loci, a double mutant was constructed between arc1, a mutant possessing many small chloroplasts, and arc5. A second double mutant was also constructed between arc3, a previously discovered mutant also possessing few large chloroplasts per cell, and arc1. Analysis of these double mutants shows that chloroplast number per mesophyll cell is greater when arc5 and arc3 mutations are expressed in the arc1 background than when expressed alone. The cell-specific nature of arc mutants was also analyzed. The phenotypic traits characteristic of arc3 and arc5 are a reduction in chloroplast number and an increase in chloroplast size in mesophyll cells: these changes are also observed in reduced form in the epidermal and guard cell chloroplasts of arc3 and arc5 plants. Analysis of parenchyma sheath cell chloroplasts suggests that in leaves of arc1 plants the normal developmental distinction between mesophyll and parenchyma sheath chloroplasts is perturbed. The relevance of these findings to the analysis of the control of chloroplast division in mesophyll cells is discussed.

Recognition of Peroxisomes by Immunofluorescence in Transformed and Untransformed Tobacco Cells.

We report the visualization of peroxisomes in tobacco (Nicotiana tabacum) leaves using fluorescently labeled antibodies to glycolate oxidase. In transgenic tobacco leaves the expression of isocitrate lyase was also visualized. In dual probing experiments both enzymes were shown to be present together in all peroxisomes in transgenic tobacco leaves.

Subcellular Visualization of Gene Transcripts Encoding Key Proteins of the Chlorophyll Accumulation Process in Developing Chloroplasts.

The coordination of the synthesis of chlorophyll (Chl) and light-harvesting Chl proteins was determined by observing the sequence of appearance of the specific mRNAs for the nuclear genes CHLH, Por, and Lhcb1*2 (AB180). CHLH encodes a magnesium protoporphyrin chelatase subunit that is involved in the first committed step in Chl biosynthesis; Por encodes protochlorophyllide oxidoreductase, which catalyzes the penultimate and only light-dependent step in Chl biosynthesis; and Lhcb1*2 encodes light-harvesting Chl a/b binding protein of the type-1 light-harvesting complex of photosystem II. Using digoxigenin-labeled antisense and sense RNA probes and a highly sensitive in situ hybridization technique, we have visualized the first appearance of the specific mRNAs in postmitotic mesophyll cells of developing 7-d-old wheat leaves (Triticum aestivum cv Maris dove). The transcripts for CHLH and POR are detectable in the youngest (18 h postmitotic) leaf tissue containing dividing cells; light-harvesting complex of photosystem II transcripts appear 12 h later. This is consistent with a requirement for accumulation of Chl before synthesis of Chl a/b binding protein can proceed at a high rate. All of the transcripts are most abundant in mesophyll cells. In the first leaf the POR message is initially restricted to the palisade, but 12 h later it is also present in the spongy mesophyll cells. All three transcripts aggregated around the surface of the chloroplasts, suggesting that translation may occur preferentially in the vicinity of the target organelle for the primary translation products.

Mitochondria Increase Three-Fold and Mitochondrial Proteins and Lipid Change Dramatically in Postmeristematic Cells in Young Wheat Leaves Grown in Elevated CO2.

A dramatic stimulation in mitochondrial biogenesis during the very early stages of leaf development was observed in young wheat plants (Triticum aestivum cv Hereward) grown in elevated CO2 (650 [mu]L L-1). An almost 3-fold increase in the number of mitochondria was observed in the very young leaf cells at the base of the first leaf of a 7-d-old wheat plant. In the same cells large increases in the accumulation of a mitochondrial chaperonin protein and the mitochondrial 2-oxoglutarate dehydrogenase complex and pyruvate dehydrogenase complex were detected by immunolabeling. Furthermore, the basal segment also shows a large increase in the rate of radiolabeling of diphosphatidylglycerol, a lipid confined to the inner mitochondrial membrane. This dramatic response in very young leaf cells to elevated CO2 suggests that the numerous documented positive effects of elevated CO2 on wheat leaf development are initiated as early as 12 h postmitosis.

arc6, A Fertile Arabidopsis Mutant with Only Two Mesophyll Cell Chloroplasts.

A novel mutant of Arabidopsis thaliana, arc6 (accumulation and replication of chloroplasts), has been isolated from a transfer DNA-mutagenized population of Arabidopsis seedlings. arc6 has the most extreme arc mutant phenotype we have yet described, with only one to three chloroplasts per leaf mesophyll cell compared to a mean of 83 in cells of the wild-type var Wassilewskija. The chloroplasts of arc6 are 20-fold larger than wild-type chloroplasts.Chloroplast division is almost certainly precluded in arc6 mesophyll cells, since chloroplast number per cell does not increase during mesophyll cell expansion. arc6 chloroplasts are long and thin in cross-section and only one-half the width of wild-type chloroplasts and the arrangement of thylakoid membranes is largely unaltered. arc6 segregates as a monogenic recessive nuclear mutation in a normal Mendelian manner and the arc6 phenotype is stably inherited for at least four generations. arc6 plants grow normally and are fertile, although the rosette leaves appear curled and twisted. arc6 plants accumulate 70 to 75% of the biomass of wild type. The phenotype of this novel mutant is discussed in relation to the nature of the control of chloroplast division in leaf cells.

Clustering of children's obesity-related behaviours: associations with sociodemographic indicators.

BACKGROUND/OBJECTIVES: Research suggests obesity-related behaviours cluster together in children and adolescents, but how these cluster patterns differ by sociodemographic indicators remains unclear. Furthermore, few studies examining clustering of behaviours have included younger children or an objective measure of physical activity (PA) and sedentary behaviour. Therefore, the aim of this study was to examine clustering patterns of diet, PA and sedentary behaviour in 5- to 6- and 10- to 12-year-old children, and their cross-sectional associations with sociodemographic indicators. SUBJECTS/METHODS: In this cross-sectional study, data from the baseline wave (2002/2003) of the Health Eating and Play study (HEAPS) were used. Questionnaires were completed by parents of Australian children aged 5-6 (n=362) and 10-12 years (n=610). Children wore accelerometers for up to 7 days. K-medians cluster analysis identified groups of children with similar diet, PA and sedentary behaviours. Chi-square tests assessed cluster differences by gender, maternal education and marital status. RESULTS: For each age group, three reliable and meaningful clusters were identified and labelled 'most healthy', 'energy-dense (ED) consumers who watch TV' and 'high sedentary behaviour/low moderate-to-vigorous PA (MVPA)'. Clusters varied by sociodemographic indicators. For example, a higher proportion of older girls comprised the 'high sedentary behaviour/low MVPA' cluster (χ(2)=22.4, P<0.001). Among both age groups, the 'ED consumers who watch TV' cluster comprised more children with lower educated mothers (younger children: χ(2)=34.9, P<0.001; older children: χ(2)=27.3, P<0.001). CONCLUSIONS: Identification of cluster patterns of obesity-related risk factors in children, and across sociodemographic groups may assist the targeting of public health initiatives, to those most in need.

Stability and plasticity during chloroplast development.

Chloroplast development occurs during cellular development. In non-limiting conditions chloroplast development is a highly conserved process, it is also complex and involves the continuous interaction of both chloroplast and nuclear genomes. In the first part of the paper the sequential and structural changes characteristic of chloroplast division and development in angiosperms are described. The synthesis of the major chloroplast components including chlorophylls a and b, lipids, nucleic acids and the major soluble and membrane proteins are then described. Chloroplast development in biochemical terms is a quantitative accretion of additional functional units. In development from proplastid to fully mature chloroplast the molecular changes are almost exclusively quantitative and the youngest plastids that can be analysed are already photochemically fully competent. In the second part of the paper the dominant role of the nuclear genome in chloroplast development is discussed. Recent work in the author's laboratory on the synthesis and accumulation of ribulose bisphosphate carboxylase-oxygenase in the developing chloroplasts of young wheat leaves is cited to illustrate the stable genomic and genotypic differences that can be recognized. In comparisons of wheat species of differing ploidy, in hexaploid cultivars and in artificially processed genetic lines, several genomic and genotypic effects have been detected. The possibilities for future investigation are discussed.

Rapid Image Analysis Screening Procedure for Identifying Chloroplast Number Mutants in Mesophyll Cells of Arabidopsis thaliana (L.) Heynh.

To analyze the genetic control of the process of chloroplast division, a direct image analysis screening procedure has been developed in which mutants of Arabidopsis thaliana (L.) Heynh. var Landsberg erecta are selected on the basis of abnormal chloroplast number. The selection procedure is based on image analysis thresholding after iodine staining, which facilitates the automatic counting of chloroplasts in isolated mesophyll cells. M2 seedlings are screened for significant deviation from the wild type relationship between mesophyll cell size and chloroplast number. Mutants with both abnormally high and abnormally low chloroplast numbers were identified. Of 3500 individual M2 seedlings screened, 18 mutant lines have been isolated and shown to be stably inherited in three subsequent generations. The most extreme phenotypes show an 80% reduction or a 50% increase in chloroplast number per mesophyll cell.

The effect of adenosine 5'-triphosphate on the shibata shift and on associated structural changes in the conformation of the prolamellar body in isolated maize etioplasts.

Isolated maize (Zea mays var. kelvedon glory) etioplasts have been used to investigate the relationships between the spectral shifts and ultrastructural changes which occur during light-induced chloroplast development. After primary photoconversion, the Shibata shift was observed as a change from 680 to 670 nm in the chlorophyllide absorption maximum. When 1.5 nm ATP was added to the incubation medium the maximum was 675 nm even after 3.5 hours of illumination. Difference spectra for this effect indicate ATP inhibition of the Shibata shift. Two bands with maxima at 682 and 669 nm can be used to fit spectra of both ATP-treated and control etioplasts, the estimated proportions of chlorophyllide 682 being 36% and 6%, respectively. Quantitative analysis of electron micrographs of the etioplasts showed that the frequency of untransformed prolamellar bodies was also higher in the presence of ATP (73% untransformed compared to 22% in the absence of ATP). A similar correlation was observed when transformation was measured for two etioplast fractions which show the shift to different extents. These results imply that the Shibata shift and prolamellar body transformation are related events, both being inhibited by the presence of ATP. ATP may therefore have an important role in regulating the early stages of plastid development.

The Effect of ATP on the Photoconversion of Protochlorophyllide in Isolated Etioplasts of Zea mays.

The transformation of protochlorophyllide (PChle) into chlorophyllide (Chle) has been studied in isolated etioplasts from Zea mays. ATP (1.5mm) prevented the transformation of photoconvertible PChle 650 to PChle 630 in aged etioplasts. Curve analysis indicated that the ATP effect on photoconvertibility could be entirely accounted for by changes in the proportions of PChle 630 and PChle 650 and examination of the isolated pigments revealed that only unphytylated PChle could be activated for photoconversion by ATP. In etioplasts aged for 5 hours, ATP also stimulated photoconversion of PChle 630 into Chle 670. The process was temperature-sensitive and involved PChle 650 and Chle 680 as intermediates. AMP alone had no effect, but inhibited ATP retardation of PChle loss. ADP had a similar but lesser effect than ATP. The ADP response, but not the ATP response, was considerably enhanced in the presence of an ATP-generating system (phosphoenolpyruvate/pyruvate kinase). UTP, GTP, and CTP gave 40 to 50% of the ATP response with intact etioplasts. In envelope-free etioplasts, ATP gave the greatest response but the other nucleotides were now 80% as effective as ATP. After primary photoconversion, ATP stimulated resynthesis of PChle 650. It is proposed that ATP both gives the holochrome the ability to bind to the PChle molecule and enables additional association of the pigment-protein complex to form PChle 650.

Sequential changes in the lipids of developing proplastids isolated from green maize leaves.

Changes in lipid composition were followed as a proplastid develops into a chloroplast. Methods were devised for the isolation of developing proplastids from sections of five different ages from the same 7-day-old maize (Zea mays var. Kelvedon Glory) leaf. Electron micrographs illustrate the homogeneity of the five types of plastid suspension, minimal contamination with other cytoplasmic membranes, and the presence of morphologically intact plastids in the proportions 85% (youngest), 85%, 80%, 70% and 60% (oldest), respectively. Both bundle sheath and mesophyll plastids are well preserved in isolation. Plastid numbers were determined from calibration curves of the chlorophyll content of each type of suspension, and lipid values then expressed as nmoles/10(6) plastids. Monogalactosyl diglyceride (MGDG), digalactosyl diglyceride (DGDG), sulfoquinovosyl diglyceride, and phosphatidyl glycerol (PG) all increase during plastid development but the rate of increase is different for each lipid. The largest changes are in MGDG (6-fold) and DGDG (4-fold). Phosphatidyl choline shows a continuous decline during plastid development. Phosphatidyl inositol and phosphatidyl ethanolamine were found in all the suspensions in low concentrations (0.4-4.0% of total lipid): calculations showed their presence could not be accounted for by bacterial or mitochondrial contamination. The increase in PG parallels the chlorophyll changes during development and at maturity 1 molecule of PG is present per 3 molecules of chlorophyll. The results are discussed in the context of the molecular structure of the photosynthetic thylakoid membranes.

Localization of DNA in Mature and Young Wheat Chloroplasts Using the Fluorescent Probe 4'-6-Diamidino-2-phenylindole.

The spatial organization of chloroplast DNA in developing and dividing wheat chloroplasts was studied in the light microscope using the fluorescent probe 4'-6-diamidino-2-phenylindole, which binds specifically to DNA.The DNA of wheat chloroplasts was localized at the periphery of the plastid, frequently in a discrete band. No relocalization of the DNA was observed during plastid replication. This peripheral location of the DNA was shown to differ from the central random location of DNA in tobacco and spinach chloroplasts.

Photosynthesis of Lipids from CO(2) in Spinacia oleracea.

Young expanding spinach leaves exposed to (14)CO(2) under physiological conditions for up to 20 minutes assimilated CO(2) into lipids at a mean rate of 7.6 micromoles per milligram chlorophyll per hour following a lag period of 5 minutes. Label entered into all parts of the lipid molecule and only 28% of the (14)C fixed into lipids was found in the fatty acid moieties, i.e. fatty acids were synthesized from CO(2)in vivo at a mean rate of 2.1 micromoles per milligram chlorophyll per hour. Intact spinach chloroplasts isolated from these leaves incorporated H(14)CO(3) into fatty acids at a maximal rate of 0.6 micromole per milligram chlorophyll per hour, but were unable to synthesize either the polar moieties of their lipids or polyunsaturated fatty acids. Since isolated chloroplasts will only synthesize fatty acids at rates similar to the one obtained with intact leaves in vivo if acetate is used as a precursor, it is suggested that acetate derived from leaf mitochondria is the physiological fatty acid precursor.