The Use of a Stringent Selection System Allows the Identification of DNA Elements that Augment Gene Expression

The use of high stringency selection systems often results in the induction of very few recombinant mammalian cell lines, which limits the ability to isolate a cell line with favorable characteristics. The employment of for instance STAR elements in DNA constructs elevates the induced number of colonies and also the protein expression levels in these colonies. Here, we describe a method to systematically identify genomic DNA elements that are able to induce many stably transfected mammalian cell lines. We isolated genomic DNA fragments upstream from the human Rb1 and p73 gene loci and cloned them around an expression cassette that contains a very stringent selection marker. Due to the stringency of the selection marker, hardly any colony survives without flanking DNA elements. We tested fourteen ~3500 bp DNA stretches from the Rb1 and p73 loci. Only two ~3500 bp long DNA fragments, called Rb1E and Rb1F, induced many colonies in the context of the stringent selection system and these colonies displayed high protein expression levels. Functional analysis showed that the Rb1 DNA fragments contained no enhancer, promoter, or STAR activity. Our data show the potential of a methodology to identify novel gene expression augmenting DNA elements in an unbiased manner

Ingestion of micronutrient fortified breakfast cereal has no influence on immune function in healthy children: A randomized controlled trial

<p>Abstract</p> <p>Background</p> <p>This study investigated the influence of 2-months ingestion of an "immune" nutrient fortified breakfast cereal on immune function and upper respiratory tract infection (URTI) in healthy children during the winter season.</p> <p>Methods</p> <p>Subjects included 73 children (N = 42 males, N = 31 females) ranging in age from 7 to 13 years (mean ± SD age, 9.9 ± 1.7 years), and 65 completed all phases of the study. Subjects were randomized to one of three groups--low, moderate, or high fortification--with breakfast cereals administered in double blinded fashion. The "medium" fortified cereal contained B-complex vitamins, vitamins A and C, iron, zinc, and calcium, with the addition of vitamin E and higher amounts of vitamins A and C, and zinc in the "high" group. Immune measures included delayed-typed hypersensitivity, global IgG antibody response over four weeks to pneumococcal vaccination, salivary IgA concentration, natural killer cell activity, and granulocyte phagocytosis and oxidative burst activity. Subjects under parental supervision filled in a daily log using URTI symptoms codes.</p> <p>Results</p> <p>Subjects ingested 3337 ± 851 g cereal during the 2-month study, which represented 14% of total diet energy intake and 20-85% of selected vitamins and minerals. Despite significant increases in nutrient intake, URTI rates and pre- to- post-study changes in all immune function measures did not differ between groups.</p> <p>Conclusions</p> <p>Data from this study indicate that ingestion of breakfast cereal fortified with a micronutrient blend for two winter months by healthy, growing children does not significantly influence biomarkers for immune function or URTI rates.</p

Species and abundance of ectoparasitic flies (Diptera) in pied flycatcher nests in Fennoscandia

Peer reviewe

The effect of a curriculum-based physical activity intervention on accelerometer-assessed physical activity in schoolchildren: a non-randomised mixed methods controlled before-and-after study

Classroom-based physical activity (PA) interventions offer the opportunity to increase PA without disrupting the curriculum. We aimed to explore the feasibility and potential effectiveness of a classroom-based intervention on moderate to vigorous PA (MVPA) and total PA. The secondary aim was to assess the acceptability and sustainability of the intervention. In a mixed-methods, non-randomised, exploratory controlled before-and-after study, 152 children (10 ± 0.7 years) were recruited from five schools; two intervention (n = 72) and three control (n = 80) schools. School teachers delivered an 8-week classroom-based intervention, comprising of 10 minutes daily MVPA integrated into the curriculum. The control schools maintained their usual school routine. Mean daily MVPA (min), total PA (mean cpm), physical fitness, and health-related quality of life measurements were taken at baseline, end of intervention, and 4-weeks post-intervention (follow-up). Data were analysed using a constrained baseline longitudinal analysis model accounting for the hierarchical data structure. For the primary outcomes (MVPA and total PA) the posterior mean difference and 95% compatibility interval were derived using a semi-Bayesian approach with an explicit prior. The acceptability and sustainability of the intervention was explored via thematic content analysis of focus group discussions with teachers (n = 5) and children (n = 50). The difference in mean daily MVPA (intervention-control) was 2.8 (-12.5 to 18.0) min/day at 8 weeks and 7.0 (-8.8 to 22.8) min/day at follow-up. For total PA, the differences were -2 (-127 to 124) cpm at 8-weeks and 11 (-121 to 143) cpm at follow-up. The interval estimates indicate that meaningful mean effects (both positive and negative) as well as trivial effects are reasonably compatible with the data and design. The intervention was received positively with continuation reported by the teachers and children. Classroom-based PA could hold promise for increasing average daily MVPA, but a large cluster randomised controlled trial is required

Evaluation of a Rapid Immunochromatographic ODK-0901 Test for Detection of Pneumococcal Antigen in Middle Ear Fluids and Nasopharyngeal Secretions

Since the incidence of penicillin-resistant Streptococcus pneumoniae has been increasing at an astonishing rate throughout the world, the need for accurate and rapid identification of pneumococci has become increasingly important to determine the appropriate antimicrobial treatment. We have evaluated an immunochromatographic test (ODK-0901) that detects pneumococcal antigens using 264 middle ear fluids (MEFs) and 268 nasopharyngeal secretions (NPSs). A sample was defined to contain S. pneumoniae when optochin and bile sensitive alpha hemolytic streptococcal colonies were isolated by culture. The sensitivity and specificity of the ODK-0901 test were 81.4% and 80.5%, respectively, for MEFs from patients with acute otitis media (AOM). In addition, the sensitivity and specificity were 75.2% and 88.8%, respectively, for NPSs from patients with acute rhinosinusitis. The ODK-0901 test may provide a rapid and highly sensitive evaluation of the presence of S. pneumoniae and thus may be a promising method of identifying pneumococci in MEFs and NPSs

The combined effect of determinants on coverage of intermittent preventive treatment of malaria during pregnancy in the Kilombero Valley, Tanzania

BACKGROUND\ud \ud Intermittent preventive treatment during pregnancy (IPTp) at routine antenatal care (ANC) clinics is an important and efficacious intervention to reduce adverse health outcomes of malaria infections during pregnancy. However, coverage for the recommended two IPTp doses is still far below the 80% target in Tanzania. This paper investigates the combined impact of pregnant women's timing of ANC attendance, health workers' IPTp delivery and different delivery schedules of national IPTp guidelines on IPTp coverage.\ud \ud METHODS\ud \ud Data on pregnant women's ANC attendance and health workers' IPTp delivery were collected from ANC card records during structured exit interviews with ANC attendees and through semi-structured interviews with health workers in south-eastern Tanzania. Women's timing of ANC visits and health worker's timing of IPTp delivery were analyzed in relation to the different national IPTp schedules and the outcome on IPTp coverage was modelled.\ud \ud RESULTS\ud \ud Among all women eligible for IPTp, 79% received a first dose of IPTp and 27% were given a second dose. Although pregnant women initiated ANC attendance late, their timing was in line with the national guidelines recommending IPTp delivery between 20-24 weeks and 28-32 weeks of gestation. Only 15% of the women delayed to the extent of being too late to be eligible for a first dose of IPTp. Less than 1% of women started ANC attendance after 32 weeks of gestation. During the second IPTp delivery period health workers delivered IPTp to significantly less women than during the first one (55% vs. 73%) contributing to low second dose coverage. Simplified IPTp guidelines for front-line health workers as recommended by WHO could lead to a 20 percentage point increase in IPTp coverage.\ud \ud CONCLUSIONS\ud \ud This study suggests that facility and policy factors are greater barriers to IPTp coverage than women's timing of ANC attendance. To maximize the benefit of the IPTp intervention, revision of existing guidelines is needed. Training on simplified IPTp messages should be consolidated as part of the extended antenatal care training to change health workers' delivery practices and increase IPTp coverage. Pregnant women's knowledge about IPTp and the risks of malaria during pregnancy should be enhanced as well as their ability and power to demand IPTp and other ANC services

A Fragment of the LG3 Peptide of Endorepellin Is Present in the Urine of Physically Active Mining Workers: A Potential Marker of Physical Activity

Biomarker analysis has been implemented in sports research in an attempt to monitor the effects of exertion and fatigue in athletes. This study proposed that while such biomarkers may be useful for monitoring injury risk in workers, proteomic approaches might also be utilised to identify novel exertion or injury markers. We found that urinary urea and cortisol levels were significantly elevated in mining workers following a 12 hour overnight shift. These levels failed to return to baseline over 24 h in the more active maintenance crew compared to truck drivers (operators) suggesting a lack of recovery between shifts. Use of a SELDI-TOF MS approach to detect novel exertion or injury markers revealed a spectral feature which was associated with workers in both work categories who were engaged in higher levels of physical activity. This feature was identified as the LG3 peptide, a C-terminal fragment of the anti-angiogenic/anti-tumourigenic protein endorepellin. This finding suggests that urinary LG3 peptide may be a biomarker of physical activity. It is also possible that the activity mediated release of LG3/endorepellin into the circulation may represent a biological mechanism for the known inverse association between physical activity and cancer risk/survival

Nestedness of Ectoparasite-Vertebrate Host Networks

Determining the structure of ectoparasite-host networks will enable disease ecologists to better understand and predict the spread of vector-borne diseases. If these networks have consistent properties, then studying the structure of well-understood networks could lead to extrapolation of these properties to others, including those that support emerging pathogens. Borrowing a quantitative measure of network structure from studies of mutualistic relationships between plants and their pollinators, we analyzed 29 ectoparasite-vertebrate host networks—including three derived from molecular bloodmeal analysis of mosquito feeding patterns—using measures of nestedness to identify non-random interactions among species. We found significant nestedness in ectoparasite-vertebrate host lists for habitats ranging from tropical rainforests to polar environments. These networks showed non-random patterns of nesting, and did not differ significantly from published estimates of nestedness from mutualistic networks. Mutualistic and antagonistic networks appear to be organized similarly, with generalized ectoparasites interacting with hosts that attract many ectoparasites and more specialized ectoparasites usually interacting with these same “generalized” hosts. This finding has implications for understanding the network dynamics of vector-born pathogens. We suggest that nestedness (rather than random ectoparasite-host associations) can allow rapid transfer of pathogens throughout a network, and expand upon such concepts as the dilution effect, bridge vectors, and host switching in the context of nested ectoparasite-vertebrate host networks