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

Effectiveness of a web- and mobile phone-based intervention to promote physical activity and healthy eating in middle-Aged males: Randomized controlled trial of the manup study

Background: The high number of adult males engaging in low levels of physical activity and poor dietary practices, and the health risks posed by these behaviours, necessitate broad-reaching intervention strategies. IT-based (web and mobile phone) interventions can be accessed by large numbers of people, yet there are few reported IT-based interventions targeting males’ physical activity and dietary practices. Objective: This study examines the effectiveness of a 9-month IT-based intervention to improve the physical activity, dietary behaviours and health literacy in middle-aged males compared to a print-based intervention. Methods: Participants, recruited offline (e.g. newspaper ads), were randomized into either an IT-based or print-based intervention arm on a 2:1 basis in favour of the fully automated IT-based arm. Participants were adult males aged 35-54 years living in two regional cities in Queensland Australia who could access the internet, owned a mobile phone and were able to increase their activity level. The intervention, ManUp, was informed by social cognitive and self regulation theories and was specifically designed to target males. Educational materials were provided and self-monitoring of physical activity and nutrition behaviours was promoted. Intervention content was the same in both intervention arms, only the delivery mode differed, and content could be accessed throughout the 9-month study period. Participants’ physical activity, dietary behaviours, and health literacy were measured using online surveys at baseline, 3 months and 9 months. Results: A total of 301 participants completed baseline assessments, 205 in the ITbased arm and 96 in the print-based arm. A total of 124 participants completed all three assessments. There were no significant between group differences in physical 5 activity and dietary behaviours (p ≥0.05). Participants reported an increased number of minutes and sessions of physical activity at 3 months (b(exp)=1.45, 95% CI=1.09-1.95; b(exp)=1.61, 95% CI=1.17-2.22) and 9 months (b(exp)=1.55, 95% CI=1.14-2.10; b(exp)=1.51, 95% CI=1.15-2.00). Overall dietary behaviours improved at 3 months (b(exp)=1.07, 95% CI=1.03-1.11) and 9 months (b(exp)=1.10, 95% CI=1.05-1.13). The proportion of participants in both groups eating higher-fibre bread and low-fat milk increased at 3 months (b(exp) = 2.25, 95% CI = 1.29-3.92; b(exp)=1.65, 95% CI = 1.07-2.55). Participants in the IT-based arm were less likely to report that 30 minutes of physical activity per day improves health (b(exp)=0.48, 95% CI=0.26-0.90) and more likely to report that vigorous intensity physical activity 3 times per week is essential (b(exp)=1.70, 95% CI=1.02-2.82). The average number of logins to the IT-platform at 3 and 9 months was 6.99 (SE=0.86) and 9.22 (SE=1.47), respectively. The average number of self-monitoring entries at 3 and 9 months was 16.69 (SE=2.38) and 22.51 (SE=3.79), respectively. Conclusions: The ManUp intervention was effective in improving physical activity and dietary behaviours in middle aged males with no significant differences between IT- and print-based delivery modes

Oligonucleotide Microarray Analysis of Age-Related Gene Expression Profiles in Miniature Pigs

Miniature pigs are useful model animals for humans because they have similar anatomy and digestive physiology to humans and are easy to breed and handle. In this study, whole blood microarray analyses were conducted to evaluate variations of correlation among individuals and ages using specific pathogen-free (SPF) Clawn miniature pigs. Whole blood RNA is easy to handle compared to isolated white blood cell RNA and can be used for health and disease monitoring and animal control. In addition, whole blood is a heterogeneous mixture of subpopulation cells. Once a great change occurs in composition and expressing condition of subpopulations, their associated change will be reflected on whole blood RNA. From 12 to 30 weeks of age, fractions of lymphocytes, monocytes, neutrophils, eosinophils, and basophils in white blood cells showed insignificant differences with age as a result of ANOVA analysis. This study attempted to identify characteristics of age-related gene expression by taking into account the change in the number of expressed genes by age and similarities of gene expression intensity between individuals. As a result, the number of expressed genes was less in fetal stage and infancy period but increased with age, reaching a steady state of gene expression after 20 weeks of age. Variation in gene expression intensity within the same age was great in fetal stage and infancy period, but converged with age. The variation between 20 and 30 weeks of age was comparable to that among 30 weeks individuals. These results indicate that uniformity of laboratory animals is expected for miniature pigs after 20 weeks of age. Furthermore, a possibility was shown that whole blood RNA analysis is applicable to evaluation of physiological state

Inter-laboratory reproducibility of fast gas chromatography–electron impact–time of flight mass spectrometry (GC–EI–TOF/MS) based plant metabolomics

The application of gas chromatography–mass spectrometry (GC–MS) to the ‘global’ analysis of metabolites in complex samples (i.e. metabolomics) has now become routine. The generation of these data-rich profiles demands new strategies in data mining and standardisation of experimental and reporting aspects across laboratories. As part of the META-PHOR project’s (METAbolomics for Plants Health and OutReach: http://www.meta-phor.eu/) priorities towards robust technology development, a GC–MS ring experiment based upon three complex matrices (melon, broccoli and rice) was launched. All sample preparation, data processing, multivariate analyses and comparisons of major metabolite features followed standardised protocols, identical models of GC (Agilent 6890N) and TOF/MS (Leco Pegasus III) were also employed. In addition comprehensive GC×GC–TOF/MS was compared with 1 dimensional GC–TOF/MS. Comparisons of the paired data from the various laboratories were made with a single data processing and analysis method providing an unbiased assessment of analytical method variants and inter-laboratory reproducibility. A range of processing and statistical methods were also assessed with a single exemplary dataset revealing near equal performance between them. Further investigations of long-term reproducibility are required, though the future generation of global and valid metabolomics databases offers much promise

Inflammation and breast cancer. Metalloproteinases as common effectors of inflammation and extracellular matrix breakdown in breast cancer

Two rapidly evolving fields are converging to impact breast cancer: one has identified novel substrates of metalloproteinases that alter immune cell function, and the other has revealed a role for inflammation in human cancers. Evidence now shows that the mechanisms underlying these two fields interact in the context of breast cancer, providing new opportunities to understand this disease and uncover novel therapeutic strategies. The metalloproteinase class of enzymes is well studied in mammary gland development and physiology, but mostly in the context of extracellular matrix modification. Aberrant metalloproteinase expression has also been implicated in breast cancer progression, where these genes act as tumor modifiers. Here, we review how the metalloproteinase axis impacts mammary physiology and tumorigenesis and is associated with inflammatory cell influx in human breast cancer, and evaluate its potential as a regulator of inflammation in the mammary gland