HPV-18 transformed cells fail to arrest in G1 in response to quercetin treatment

Previous work with primary human keratinocytes demonstrated that quercetin, a potent mutagen found in high levels in bracken fern (Pteridium aquilinum), arrested cells in G1 with concomitant elevation of the cyclin-dependent kinase inhibitor (cdki) p27Kip1. Expression of the human papillomavirus type 16 (HPV-16) E6 and E7 oncoproteins, under transcriptional control of a heterologous promoter, in transformed keratinocytes failed to abrogate this arrest [Beniston, R., Campo, M.S., 2003. Quercetin elevates p27(Kip1) and arrests both primary and HPV-16 E6/E7 transformed human keratinocytes in G1. Oncogene 22, 5504–5514]. Given the link between papillomavirus infection, bracken fern in the diet and cancer of the oesophagus in humans, we wished to investigate further whether cells transformed by the whole genome of HPV-16 or HPV-18, with E6 and E7 under the transcriptional control of their respective homologous promoters, would be similarly arrested in G1 by quercetin. In agreement with earlier work, quercetin arrested HPV-16 transformed cells in G1 with an increase in the cyclin-dependent kinase inhibitor p27Kip1. However, HPV-18 transformed cells did not arrest after quercetin treatment. The failure of HPV-18 transformed cells to arrest in G1 was linked to the up-regulation of the HPV-18 long control region (LCR) by quercetin, maintaining high expression of the viral transforming proteins. Transcriptional up-regulation of the HPV-18 LCR was mediated by a “quercetin responsive element” homologous to the one identified previously in the bovine papillomavirus type 4 (BPV-4) LCR

Quercetin elevates p27Kip1 and arrests both primary and HPV16 E6/E7 transformed human keratinocytes in G1

Our previous work with primary bovine fibroblasts demonstrated that quercetin, a potent mutagen found in high levels in bracken fern (Pteridium aquilinum), arrested cells in G1 and G2/M, in correlation with p53 activation. The expression of bovine papillomavirus type 4 (BPV-4) E7 overcame this arrest and lead to the development of tumorigenic cells lines (Beniston et al., 2001). Given the possible link between papillomavirus infection, bracken fern in the diet and cancer of the upper gastrointestinal (GI) tract in humans, we investigated whether a similar situation would occur in human cells transformed by human papillomavirus type 16 (HPV-16) oncoproteins. Quercetin arrested primary human foreskin keratinocytes in G1. Arrest was linked to an elevation of the cyclin-dependent kinase inhibitor (cdki) p27Kip1. Expression of the HPV16 E6 and E7 oncoproteins in transformed cells failed to abrogate cell cycle arrest. G1 arrest in the transformed cells was also linked to an increase of p27Kip1 with a concomitant reduction of cyclin E-associated kinase activity. This elevation of p27Kip1 was due not only to increased protein half-life, but also to increased mRNA transcription

Optical ‘dampening’ of the refractive error to axial length ratio:implications for outcome measures in myopia control studies

Purpose: To gauge the extent to which differences in the refractive error axial length relationship predicted by geometrical optics are observed in actual refractive/biometric data.Methods: This study is a retrospective analysis of existing data. Right eye refractive error [RX] and axial length [AXL] data were collected on 343 6-to-7-year-old children [mean 7.18 years (SD 0.35)], 294 12-to-13-year-old children [mean 13.12 years (SD 0.32)] and 123 young adults aged 18-to-25-years [mean 20.56 years (SD 1.91)]. Distance RX was measured with the Shin-Nippon NVision-K 5001 infrared open-field autorefractor. Child participants were cyclopleged prior to data collection (1% Cyclopentolate Hydrochloride). Myopia was defined as a mean spherical equivalent [MSE] ≤-0.50D. Axial length was measured using the Zeiss IOLMaster 500. Optical modelling was based on ray tracing and manipulation of parameters of a Gullstrand reduced model eye.Results: There was a myopic shift in mean MSE with age (6-7 years +0.87 D, 12-13 years -0.06 D and 18-25 years -1.41 D), associated with an increase in mean AXL (6-7 years 22.70 mm, 12-13 years 23.49 mm and 18-25 years 23.98 mm). There was a significant negative correlation between MSE and AXL for all age groups (all p <0.005). RX: AXL ratios for participant data were compared with the ratio generated from Gullstrand model eyes. Both modelled and actual data showed non-linearity and non-constancy, and that as axial length is increased, the relationship between myopia and axial length differs, such that it becomes more negative.Conclusions: Optical theory predicts that there will be a reduction in the RX: AXL ratio with longer eyes. The participant data although adhering to this theory show a reduced effect, with eyes with longer axial lengths having a lower refractive error to axial length ratio than predicted by model eye calculations. We propose that in myopia control intervention studies when comparing efficacy, consideration should be given to the dampening effect seen with a longer eye

Six Year Refractive Change among White Children and Young Adults: Evidence for Significant Increase in Myopia among White UK Children

OBJECTIVE:To determine six-year spherical refractive error change among white children and young adults in the UK and evaluate differences in refractive profiles between contemporary Australian children and historical UK data. DESIGN:Population-based prospective study. PARTICIPANTS:The Northern Ireland Childhood Errors of Refraction (NICER) study Phase 1 examined 1068 children in two cohorts aged 6-7 years and 12-13 years. Prospective data for six-year follow-up (Phase 3) are available for 212 12-13 year olds and 226 18-20 year olds in each cohort respectively. METHODS:Cycloplegic refractive error was determined using binocular open-field autorefraction (Shin-Nippon NVision-K 5001, cyclopentolate 1%). Participants were defined by spherical equivalent refraction (SER) as myopic SER ≤-0.50D, emmetropic -0.50D<SER<+2.00 or hyperopic SER≥+2.00D. MAIN OUTCOME MEASURES:Proportion and incidence of myopia. RESULTS:The proportion of myopes significantly increased between 6-7 years (1.9%) and 12-13 years (14.6%) (p<0.001) but not between 12-13 and 18-20 years (16.4% to 18.6%, p = 0.51). The estimated annual incidence of myopia was 2.2% and 0.7% for the younger and older cohorts respectively. There were significantly more myopic children in the UK at age 12-13 years in the NICER study (16.4%) than reported in Australia (4.4%) (p<0.001). However by 17 years the proportion of myopia neared equivalence in the two populations (NICER 18.6%, Australia 17.7%, p = 0.75). The proportion of myopic children aged 12-13 years in the present study (2006-2008) was 16.4%, significantly greater than that reported for children aged 10-16 years in the 1960's (7.2%, p = 0.01). The proportion of hyperopes in the younger NICER cohort decreased significantly over the six year period (from 21.7% to 14.2%, p = 0.04). Hyperopes with SER ≥+3.50D in both NICER age cohorts demonstrated persistent hyperopia. CONCLUSIONS:The incidence and proportion of myopia are relatively low in this contemporary white UK population in comparison to other worldwide studies. The proportion of myopes in the UK has more than doubled over the last 50 years in children aged between 10-16 years and children are becoming myopic at a younger age. Differences between the proportion of myopes in the UK and in Australia apparent at 12-13 years were eliminated by 17 years of age

Evaluation of a commercial web-based weight loss and weight loss maintenance program in overweight and obese adults: a randomized controlled trial

<p>Abstract</p> <p>Background</p> <p>Obesity rates in adults continue to rise and effective treatment programs with a broad reach are urgently required. This paper describes the study protocol for a web-based randomized controlled trial (RCT) of a commercially available program for overweight and obese adult males and females. The aim of this RCT was to determine and compare the efficacy of two web-based interventions for weight loss and maintenance of lost weight.</p> <p>Methods/Design</p> <p>Overweight and obese adult males and females were stratified by gender and BMI and randomly assigned to one of three groups for 12-weeks: waitlist control, or basic or enhanced online weight-loss. Control participants were re-randomized to the two weight loss groups at the end of the 12-week period. The basic and enhanced group participants had an option to continue or repeat the 12-week program. If the weight loss goal was achieved at the end of 12, otherwise on completion of 24 weeks of weight loss, participants were re-randomized to one of two online maintenance programs (maintenance basic or maintenance enhanced), until 18 months from commencing the weight loss program. Assessments took place at baseline, three, six, and 18 months after commencing the initial weight loss intervention with control participants repeating the initial assessment after three month of waiting. The primary outcome is body mass index (BMI). Other outcomes include weight, waist circumference, blood pressure, plasma markers of cardiovascular disease risk, dietary intake, eating behaviours, physical activity and quality of life.</p> <p>Both the weight loss and maintenance of lost weight programs were based on social cognitive theory with participants advised to set goals, self-monitor weight, dietary intake and physical activity levels. The enhanced weight loss and maintenance programs provided additional personalized, system-generated feedback on progress and use of the program. Details of the methodological aspects of recruitment, inclusion criteria, randomization, intervention programs, assessments and statistical analyses are described.</p> <p>Discussion</p> <p>Importantly, this paper describes how an RCT of a currently available commercial online program in Australia addresses some of the short falls in the current literature pertaining to the efficacy of web-based weight loss programs.</p> <p>Australian New Zealand Clinical Trials Registry (ANZCTR) number: ACTRN12610000197033</p

Factors associated with early childhood education and care service implementation of healthy eating and physical activity policies and practices in Australia: a cross-sectional study

Many early childhood education and care (ECEC) services fail to implement recommended policies and practices supportive of healthy eating and physical activity. The purpose of this study was to assess whether certain theoretically-based factors are associated with implementation of healthy eating and physical activity policies and practices in a sample of ECEC services. A cross-sectional survey was conducted with Service Managers of ECEC services. The survey assessed the operational characteristics, policy, and practice implementation, and 13 factors were suggested by Damschroder's Consolidated Framework for Implementation Research to impede or promote implementation. Logistic regression analyses found a significant association between implementation factor score and full implementation (OR 1.38; 95% CI 1.18-1.61; <i>p</i> = <0.01), indicating that for every one point increase in implementation score, ECEC services were 38% more likely to be fully implementing the policies and practices. The findings highlight the opportunities for improving implementation of obesity prevention interventions in this setting by developing interventions that address such factors

Validity of a measure to assess healthy eating and physical activity policies and practices in Australian childcare services

Childcare services represent a valuable obesity prevention opportunity, providing access to a large portion of children at a vital point in their development. Few rigorously validated measures exist to measure healthy eating and physical activity policies and practices in this setting, and no such measures exist that are specific to the childcare setting in Australia. Methods. This was a cross sectional study, comparing two measures (pen and paper survey and observation) of healthy eating and physical activity policies and practices in childcare services. Research assistants attended consenting childcare services (n = 42) across the Hunter region of New South Wales, Australia and observed practices for one day. Nominated Supervisors and Room Leaders of the service also completed a pen and paper survey during the day of observation. Kappa statistics and proportion agreement were calculated for a total of 43 items relating to healthy eating and physical activity policies and practices. Results: Agreement ranged from 38%-100%. Fifty one percent of items showed agreement of greater than or equal to 80%. Items assessing the frequency with which staff joined in active play with children reported the lowest percent agreement, while items assessing availability of beverages such as juice, milk and cordial, as well as the provision of foods such as popcorn, pretzels and sweet biscuits, reported the highest percent agreement. Kappa scores ranged from -0.06 (poor agreement) to 1 (perfect agreement). Of the 43 items assessed, 27 were found to have moderate or greater agreement. Conclusions: The study found that Nominated Supervisors and Room Leaders were able to accurately report on a number of healthy eating and physical activity policies and practices. Items assessing healthy eating practices tended to have higher kappa scores than those assessing physical activity related policies or practices. The tool represents a useful instrument for public health researchers and policy makers working in this setting

A randomised controlled trial of an intervention to facilitate the implementation of healthy eating and physical activity policies and practices in childcare services.

Childhood overweight and obesity tracks into adulthood, increasing the risk of developing future chronic disease. Implementing initiatives promoting healthy eating and physical activity in childcare settings has been identified as a priority to prevent excessive child weight gain. Despite this, few trials have been conducted to assess the effectiveness of interventions to support population-wide implementation of such initiatives. The aim of this study is to assess the effectiveness of a multicomponent intervention in increasing the implementation of healthy eating and physical activity policies and practices by centre-based childcare services