Gathering Perspectives on Extended Family Influence on African American Children\u27s Physical Activity

Background: The family environment is a key determinant of children’s physical activity. The importance of the extended African American family is well established, but there is little research on its influence on school age children’s physical activity. Methods: We recruited eight families in which grandparents and other adult relatives played a central role in child supervision. Semi-structured interviews with parents, other adult relatives, and children revealed various perspectives on the influences of culture and families on children’s weight-related behaviors. Results: Children were between the ages of 6 and 11, and five of the families resided in neighborhoods in which at least 20% of the households reported a total income below the poverty level. Adults described efforts to develop active lifestyles for their families. Children talked about experiencing different types of behavioral influences from parents and other adult relatives. Conclusions: The findings suggest that resource limitations, neighborhood characteristics, and adults’ perceptions of children’s activity needs influence the ways that parents and adult relatives engage children in physical activity

Lipid rafts are disrupted in mildly inflamed intestinal microenvironments without overt disruption of the epithelial barrier

Intestinal epithelial barrier disruption is a feature of inflammatory bowel disease (IBD), but whether barrier disruption precedes or merely accompanies inflammation remains controversial. Tight junction (TJ) adhesion complexes control epithelial barrier integrity. Since some TJ proteins reside in cholesterol-enriched regions of the cell membrane termed lipid rafts, we sought to elucidate the relationship between rafts and intestinal epithelial barrier function. Lipid rafts were isolated from Caco-2 intestinal epithelial cells primed with the proinflammatory cytokine interferon-γ (IFN-γ) or treated with methyl-β-cyclodextrin as a positive control for raft disruption. Rafts were also isolated from the ilea of mice in which colitis had been induced in conjunction with in vivo intestinal permeability measurements, and lastly from intestinal biopsies of ulcerative colitis (UC) patients with predominantly mild or quiescent disease. Raft distribution was analyzed by measuring activity of the raft-associated enzyme alkaline phosphatase and by performing Western blot analysis for flotillin-1. Epithelial barrier integrity was estimated by measuring transepithelial resistance in cytokine-treated cells or in vivo permeability to fluorescent dextran in colitic mice. Raft and nonraft fractions were analyzed by Western blotting for the TJ proteins occludin and zonula occludens-1 (ZO-1). Our results revealed that lipid rafts were disrupted in IFN-γ-treated cells, in the ilea of mice with subclinical colitis, and in UC patients with quiescent inflammation. This was not associated with a clear pattern of occludin or ZO-1 relocalization from raft to nonraft fractions. Significantly, a time-course study in colitic mice revealed that disruption of lipid rafts preceded the onset of increased intestinal permeability. Our data suggest for the first time that lipid raft disruption occurs early in the inflammatory cascade in murine and human colitis and, we speculate, may contribute to subsequent disruption of epithelial barrier function.Journal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

The <i>C</i>. <i>elegans</i> ortholog of Tectonic1 is a transition zone component.

<p>(A) Schematic of the X-box sequence and <i>ok3021</i> allele of <i>C</i>. <i>elegans tctn-1</i>. (B) GFP-tagged TCTN-1 localizes specifically to the transition zones (TZs) in head (amphid) and tail (phasmid) cilia of <i>C</i>. <i>elegans</i>. Basal bodies (BBs) and ciliary axonemes are marked with tdTomato-tagged XBX-1, a component of the ciliary dynein. Schematics illustrate the position of TCTN-1 at the transition zone with respect to the basal body and axoneme. Scale bar, 5 μm.</p

<i>C</i>. <i>elegans tctn-1</i> interacts with NPHP genes, but not MKS genes to affect ciliary structure.

<p>(A) Dye filling of amphid neurons in L4 nematodes in the indicated single transition zone mutants (left column) and <i>tctn-1</i> double mutants (right column). Lateral views, with anterior to the left. Genotypes including an allele affecting a previously recognized MKS complex component are indicated in green. Genotypes including an allele affecting an NPHP complex component are indicated in red. Scale bar, 20 μm. (B and C) Fluorescence intensity of DiI filled amphid neurons in single mutants of the MKS complex or NPHP complex, and double mutants with <i>tctn-1</i> relative to wild type. Error bars represent the standard deviation. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.001; ** <i>p</i><0.001 compared to <i>tctn-1</i>). (D) Low and high magnification TEM cross-sections of the distal segment, middle segment, transition zone (TZ), and distal dendritic periciliary membrane compartment (PCMC) of amphid cilia with schematics below (lateral and transverse views). Green arrowheads indicate intact Y-links in wild type and <i>tctn-1</i> transition zones whereas red arrowheads indicate reduced or missing Y-links, observed in <i>nphp-4</i> and <i>tctn-1; nphp-4</i> transition zones. Yellow arrowheads indicate open B-tubules and purple arrows indicate vesicle accumulation in the PCMCs of <i>nphp-4</i> and <i>tctn-1; nphp-4</i> mutants. <i>tctn-1; nphp-4</i> mutants display several truncated axonemes and axonemes with fewer microtubule doublets (blue arrows) compared to wild type, <i>tctn-1</i> and <i>nphp-4</i> mutants. Boxed numbers indicate distances (μm) from the distal ciliary tips. Scale bars,100 nm.</p

TCTN-1 contributes to <i>C</i>. <i>elegans</i> ciliary gate function.

<p>(A-D) Localization of different transition zone proteins in phasmid cilia of wild type, <i>tctn-1</i>, <i>nphp-4</i>, and <i>tctn-1; nphp-4</i> mutants. Each GFP, CFP, YFP, or tdTomato-tagged transition zone protein is co-localized with a ciliary marker (CHE-13, DYF-11 or XBX-1), as indicated. Abnormal localization of MKS-5 in the <i>nphp-4</i> single and <i>tctn-1; nphp-4</i> double mutants is indicated by asterisks. (E) ARL-13::GFP localizes ectopically to the PCMC in <i>tctn-1</i>, <i>nphp-4</i>, and <i>tctn-1; nphp-4</i> mutants as indicated by asterisks, but localizes normally to the middle segment (MS) of cilia. (F) ODR-10::GFP localizes to the cilia of AWA neurons in wild type, <i>tctn-1</i>, <i>nphp-4</i>, and <i>tctn-1; nphp-4</i> mutants. Anterior is to the right. (G) TRAM-1a::tdTomato localizes only to the PCMC in wild type animals, but enters phasmid cilia in <i>tctn-1</i> mutants, as indicated by asterisks. MKS-2::GFP marks the transition zone. Scale bars, 5 μm.</p

Mouse <i>Tctn1</i> genetically interacts with NPHP complex genes, but not MKS complex genes.

<p>(A) Lateral views of wild type, single or double mutant mouse embryos of indicated genotype at E14.5. Exencephaly is apparent in the <i>Tctn1</i>^-/-<i>Nphp1</i>^-/- double mutant. Corresponding Alcian blue staining of the right forelimb (top) and hindlimb (bottom) are included, with asterisks indicating extra digits. Genes encoding components of the NPHP complex are indicated in red. Genes encoding components of the MKS complex are indicated in green. Scale bars, 1 mm. (B) Number of digits in the forelimbs and (C) hindlimbs of wild type, single or double mutant embryos of indicated genotypes. (D) Incidence of exencephaly in wild type, single or double mutants embryos of indicated genotypes. Numbers of animals analyzed for polydactyly and exencephaly are included in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005627#pgen.1005627.s005" target="_blank">S1</a> and <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005627#pgen.1005627.s006" target="_blank">S2</a> Tables.</p

Mouse Tctn1 and Nphp4 have distinct roles in transition zone composition and overlapping roles in ciliogenesis.

<p>(A) Limb bud sections (left) from E11.5 embryos were stained for acetylated tubulin (Tub^Ac, green) to mark the ciliary axonemes, γ-tubulin (red) to mark the basal bodies and DAPI (blue) to mark nuclei. Scale bar, 5 μm. Transmission electron microscopy (TEM, right) for each genotype. Scale bar, 200 nm. (B) Quantification of limb bud cilia in control, <i>Tctn1</i>^-/-<i>Nphp4</i>^n/+, and <i>Tctn1</i>^-/-<i>Nphp4</i>^n/n mutants from TEM fields of view. Error bars represent the standard error of the mean. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.05; ** <i>p</i><0.001). (C) Fibroblasts derived from E13.5 limb buds of the indicated genotypes stained for Tub^Ac (green), Arl13b (red), γ-tubulin (cyan), and DAPI (blue). White arrowheads indicate cilia in <i>Tctn1</i>^-/-<i>Nphp4</i>^n/+ and <i>Tctn1</i>^-/-<i>Nphp4</i>^n/n mutants. Scale bar, 10 μm. (D) Quantification of proportion of ciliated cultured limb bud fibroblasts. Error bars represent the standard deviation. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.05; ** <i>p</i><0.0001). (E) Immunostaining of limb bud fibroblasts for Nphp1 (red), Tub^Ac (green), and γ-tubulin (cyan). Scale bar, 1 μm.</p

Roles for mouse <i>Tctn1</i> and <i>Nphp4</i> in Hh signaling and ciliary localization of Hh pathway components.

<p>(A and B). mRNA levels of <i>Gli1</i> and <i>Ptch1</i> normalized to <i>β-actin</i> in forelimb bud cells treated with DMSO or SAG. Error bars represent standard deviations. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.05). (C) Limb bud fibroblasts treated with DMSO or SAG, then immunostained for Smo (red), Tub^Ac (blue), and γ-tubulin (green). (D) Quantitation of Smo ciliary intensity in DMSO or SAG treated limb bud fibroblasts. (E) Limb bud fibroblasts treated with DMSO or SAG, then immunostained for Gpr161 (red), Tub^Ac (blue), and γ-tubulin (green). (F) Quantitation of Gpr161 ciliary intensity in DMSO or SAG treated limb bud fibroblasts. Error bars represent standard error of the mean. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.05). Scale bars, 2 μm.</p

<i>C</i>. <i>elegans</i> NPHP genes synthetically interact with <i>bbs-5</i> to affect ciliary structure.

<p>(A) Dye filling of amphid neurons in L4 nematodes in single transition zone mutants (left column) and <i>bbs-5</i> double mutants (right column). Lateral views, with anterior to the left. Scale bar, 20 μm. (B) Fluorescence intensity of DiI filled amphid neurons relative to wild type. Genotypes including an allele affecting an MKS complex component are indicated in green. Genotypes including an allele affecting an NPHP complex component are indicated in red. Error bars represent the standard deviation. Statistical significance according to unpaired Student’s <i>t</i>-tests (* <i>p</i><0.001; ** <i>p</i><0.001 compared to <i>bbs-5</i>). (C) Low and high magnification TEM cross-sections of the distal segment, middle segment, transition zone (TZ), and PCMC of amphid cilia with schematics below (lateral and transverse views). <i>bbs-5</i> mutants display normal ciliary structures, including intact Y-links (green arrowheads). <i>bbs-5; nphp-4</i> mutant cilia display open B-tubules (yellow arrowheads), reduced Y-links (red arrowheads) and vesicle accumulation in the PCMC (purple arrows) similar to <i>nphp-4</i> cilia, but have fewer axonemes that fully extend distally in both pores. Boxed numbers indicate distances (μm) from the distal ciliary tips. Scale bars,100 nm.</p