Study protocol of a cluster randomised controlled trial investigating the effectiveness of a tailored energy balance programme for recent retirees

BACKGROUND: People in transitional life stages, such as occupational retirement, are likely to gain weight and accumulate abdominal fat mass caused by changes in physical activity and diet. Hence, retirees are an important target group for weight gain prevention programmes, as described in the present paper. METHODS/DESIGN: A systematic and stepwise approach (Intervention Mapping) is used to develop a low-intensity energy balance intervention programme for recent retirees. This one-year, low-intensity multifaceted programme aims to prevent accumulation of abdominal fat mass and general weight gain by increasing awareness of energy balance and influencing related behaviours of participants' preference. These behaviours are physical activity, fibre intake, portion size and fat consumption. The effectiveness of the intervention programme is tested in a cluster randomised controlled trial. Measurements of anthropometry, physical activity, energy intake, and related psychosocial determinants are performed at baseline and repeated at 6 months for intermediate effect, at 12 months to evaluate short-term intervention effects and at 24 months to test the sustainability of the effects. DISCUSSION: This intervention programme is unique in its focus on retirees and energy balance. It aims at increasing awareness and takes into account personal preferences of the users by offering several options for behaviour change. Moreover, the intervention programme is evaluated at short-term and long-term and includes consecutive outcome measures (determinants, behaviour and body composition)

Endo-MitoEGFP mice: a novel transgenic mouse with fluorescently marked mitochondria in microvascular endothelial cells.

Blood vessel-specific fluorescent transgenic mice are excellent tools to study the development of the vasculature and angiogenic processes. There is growing interest in the biological processes relevant to endothelial cells but limited tools exist to selectively evaluate subcellular functions of this cell type in vivo. Here, we report a novel transgenic animal model that expresses mitochondrially targeted enhanced green fluorescent protein (EGFP) via the Hb9 promoter, a homeobox transcription factor with limited known involvement in the vasculature. Random integration of the transgene, containing the entire mouse Hb9 promoter, was found to be expressed in a variety of vascularised tissues. Further inspection revealed that Mito-EGFP localizes to the endothelial cells (ECs) of a subset of microvascular blood vessels, especially in the central nervous system (CNS), heart, spleen, thymus, lymph nodes and skin. We demonstrate the utility of this novel transgenic mouse, named Endo-MitoEGFP, in the detection, imaging, and isolation of microvascular ECs and evaluation of EC mitochondrial function isolated from adult animals. These transgenic mice will be useful to studies of ECs in development, physiology, and pathology

High glycemic index and glycemic load diets as risk factors for insomnia: analyses from the Women’s Health Initiative

BackgroundPrevious studies have shown mixed results on the association between carbohydrate intake and insomnia. However, any influence that refined carbohydrates have on risk of insomnia is likely commensurate with their relative contribution to the overall diet, so studies are needed that measure overall dietary glycemic index (GI), glycemic load, and intakes of specific types of carbohydrates.ObjectiveWe hypothesized that higher GI and glycemic load would be associated with greater odds of insomnia prevalence and incidence.MethodsThis was a prospective cohort study with postmenopausal women who participated in the Women's Health Initiative Observational Study, investigating the relations of GI, glycemic load, other carbohydrate measures (added sugars, starch, total carbohydrate), dietary fiber, and specific carbohydrate-containing foods (whole grains, nonwhole/refined grains, nonjuice fruits, vegetables, dairy products) with odds of insomnia at baseline (between 1994 and 1998; n = 77,860) and after 3 y of follow-up (between 1997 and 2001; n = 53,069).ResultsIn cross-sectional and longitudinal analyses, higher dietary GI was associated with increasing odds of prevalent (fifth compared with first quintile OR: 1.11; CI: 1.05, 1.16; P-trend = 0.0014) and incident (fifth compared with first quintile OR: 1.16; CI: 1.08, 1.25; P-trend < 0.0001) insomnia in fully adjusted models. Higher intakes of dietary added sugars, starch, and nonwhole/refined grains were each associated with higher odds of incident insomnia. By contrast, higher nonjuice fruit and vegetable intakes were significantly associated with lower odds of incident insomnia. Also, higher intakes of dietary fiber, whole grains, nonjuice fruit, and vegetables were significantly associated with lower odds of prevalent insomnia.ConclusionsThe results suggest that high-GI diets could be a risk factor for insomnia in postmenopausal women. Substitution of high-GI foods with minimally processed, whole, fiber-rich carbohydrates should be evaluated as potential treatments of, and primary preventive measures for, insomnia in postmenopausal women

EGFP expression is not restricted to central nervous system, but is also expressed in vascularized tissues.

<p>(A) Schematic of p<i>Hb9</i>-MitoEGFP transgenic construct, with mitochondrial targeting sequence of Cytochrome <i>c</i> Oxidase subunit VIII. (B) RT-PCR of EGFP mRNA from a panel of tissues of transgenic (+) and non-transgenic littermates (-). Actin serves as a loading control. (C) EGFP protein levels detected via immunoblotting in a panel of tissues isolated from transgenic (+) and non-transgenic littermates (-). SOD1 serves as a loading control. <i>n</i>=3-4 animals.</p

EGFP expression is detected in ECs via flow cytometry.

<p>(A) Representative dot plot of ECs isolated from spleens of transgenic mice. The EC population was selected based on size and labelling with PECAM-1. (B) The majority of splenic ECs express EGFP and PECAM-1. (C) Western blot of brains homogenates (H), as well as soluble (S1) and insoluble (S2) fractions from lysates of ECs isolated from CNS vessels of transgenic animals and immunoblotted for EGFP and p120. <i>n</i>=3 animals.</p

EGFP is expressed within mitochondria.

<p>(A) Homogenates (H), cytosolic protein (C) and purified mitochondria (M) were collected from the brain, spinal cord and spleen of transgenic mice and probed for EGFP via immunoblot. SOD1 and VDAC serve as controls for cytosol and mitochondria, respectively. (B) Mitochondria were isolated from the spleens of transgenic and non-transgenic mice. <i>Left</i>: Mitochondria were gated according to their light scattering properties (forward scatter, FSC; side scatter, SSC). <i>Middle</i>: Gated mitochondria were stained with MitoTracker Red (MTR, black, dashed) and compared to unstained (grey, shaded) mitochondria. <i>Right</i>: MTR⁺ mitochondria from transgenic (black, unshaded) and non-transgenic (grey, shaded) mice were analyzed for EGFP expression. Data presented is representative of three independent experiments. (C) Mitochondria from transgenic (black, unshaded), and non-transgenic (grey, shaded) mice were analyzed for EGFP expression. (D) The transmembrane potential of EGFP⁺ mitochondria was assayed using TMRM. Mitochondria were left, unstained (grey, filled), treated with TMRM, basal conditions (blue), or treated with the protonophore CCCP (red). (E) Mitochondrial superoxide production of EGFP⁺ mitochondria, was assayed using MitoSOX Red. Mitochondria were left, unstained (grey, filled), treated with MitoSOX Red under basal conditions (green), or treated with the complex III inhibitor Antimycin A (orange). C, D, E are from the same sample and are representative of three independent experiments.</p