Identification and validation of housekeeping genes in brains of the desert locust Schistocerca gregaria under different developmental conditions

<p>Abstract</p> <p>Background</p> <p>To obtain reliable quantitative RT-PCR data, normalization relative to stable housekeeping genes is required. However, in practice, expression levels of 'typical' housekeeping genes have been found to vary between tissues and under different experimental conditions. To date, validation studies of reference genes in insects are extremely rare and have never been performed in locusts. In this study, putative housekeeping genes were identified in the desert locust, <it>Schistocerca gregaria </it>and two different software programs (geNorm and Normfinder) were applied to assess the stability of thesegenes.</p> <p>Results</p> <p>We have identified seven orthologs of commonly used housekeeping genes in the desert locust. The selected genes were the orthologs of actin, <it>EF1a, GAPDH, RP49, TubA1, Ubi</it>, and <it>CG13220</it>. By employing real time RT-PCR we have analysed the expression of these housekeeping genes in brain tissue of fifth instar nymphs and adults. In the brain of fifth instar nymphs geNorm indicated <it>Sg-EF1a</it>, <it>Sg-GAPDH </it>and <it>Sg-RP49 </it>as most stable genes, while Normfinder ranked <it>Sg-RP49</it>, <it>Sg-EF1a </it>and <it>Sg-ACT </it>as most suitable candidates for normalization. The best normalization candidates for gene expression studies in the brains of adult locusts were <it>Sg-EF1a, Sg-GAPDH </it>and <it>Sg-Ubi </it>according to geNorm, while Normfinder determined <it>Sg-GAPDH</it>, <it>Sg-Ubi </it>and <it>Sg-ACT </it>as the most stable housekeeping genes.</p> <p>Conclusion</p> <p>To perform transcript profiling studies on brains of the desert locust, the use of <it>Sg-RP49</it>, <it>Sg-EF1a </it>and <it>Sg-ACT </it>as reference genes is proposed for studies of fifth instar nymphs. In experiments with adult brains, however, the most preferred reference genes were <it>Sg-GAPDH</it>, <it>Sg-Ubi </it>and <it>Sg-EF1a</it>. These data will facilitate transcript profiling studies in desert locusts and provide a good starting point for the initial selection of genes for validation studies in other insects.</p

Leukocyte telomere length and diet in the apparently healthy, middle-aged Asklepios population

Telomere length is a prognostic biomarker for aging diseases. As it is unknown whether diet plays a role in these associations, we aimed to assess the impact of diet on telomere length. Moreover, given that telomere length is modulated by oxidative stress and inflammation, an additional goal was to evaluate whether the latter may mediate possible telomere - diet associations. Southern blot measured leukocyte telomere length and food frequency questionnaire data were compared for 2509 apparently healthy men and women (similar to 35 to 55 years) from the Asklepios population. No significant associations were found between telomere length and overall dietary characteristics, such as dietary diversity, quality, equilibrium, and the dietary inflammatory index. Exploratory analysis of individual dietary variables revealed that a higher daily intake of deep fried potato products was associated with shorter telomeres (P = 0.002, 151 bp per 100 g/day), also in both sexes separately. Deep fried potato product consumption was also significantly associated with C-reactive protein (P = 0.032) and uric acid (P = 0.042), but not other inflammation and oxidative stress markers. These results suggest an at most limited association between overall dietary patterns and telomere length in the general population. Nevertheless, the association between telomere length and deep fried potato product intake warrants additional research

Multibehavioural Interventions with a Focus on Specific Energy Balance-Related Behaviours Can Affect Diet Quality in Preschoolers from Six European Countries: The ToyBox-Study

The present study aimed to examine whether a multibehavioural intervention with a focus on specific energy balance-related behaviours can affect total diet quality and its four subcomponents in European preschoolers and to investigate if these intervention effects differed by socioeconomic status (SES). Parents/caregivers of 3.5 to 5.5 year-olds (n = 4968) recruited through kindergartens in six European countries within the ToyBox-study completed questionnaires on socio-demographics and a food frequency questionnaire on their preschoolers' diet. To assess intervention effects and differences by SES, multilevel repeated measures analyses were conducted. In contrast to no significant difference in total diet quality, in both the intervention and control group, the dietary quality and dietary equilibrium increased, with a larger increase in the intervention group (mean difference (quality): + 3.4%;mean difference (equilibrium): +0.9%) compared to the control group ((quality): + 1.5%;(equilibrium): + 0.2%). SES was not a significant moderator for intervention effects on total diet quality, nor for the four subcomponents. This study indicates that multibehavioural interventions with a focus on specific energy balance-related behaviours in preschoolers not only affect those targeted behaviours, but can also have more generalized effects. The ToyBox-intervention effects were similar for both lower and high SES preschoolers

Three years pilot of spinal muscular atrophy newborn screening turned into official program in Southern Belgium.

Three new therapies for spinal muscular atrophy (SMA) have been approved by the United States Food and Drug Administration and the European Medicines Agency since 2016. Although these new therapies improve the quality of life of patients who are symptomatic at first treatment, administration before the onset of symptoms is significantly more effective. As a consequence, newborn screening programs have been initiated in several countries. In 2018, we launched a 3-year pilot program to screen newborns for SMA in the Belgian region of Liège. This program was rapidly expanding to all of Southern Belgium, a region of approximately 55,000 births annually. During the pilot program, 136,339 neonates were tested for deletion of exon 7 of SMN1, the most common cause of SMA. Nine SMA cases with homozygous deletion were identified through this screen. Another patient was identified after presenting with symptoms and was shown to be heterozygous for the SMN1 exon 7 deletion and a point mutation on the opposite allele. These ten patients were treated. The pilot program has now successfully transitioned into the official neonatal screening program in Southern Belgium. The lessons learned during implementation of this pilot program are reported

Perception of inappropriate cardiopulmonary resuscitation by clinicians working in emergency departments and ambulance services : The REAPPROPRIATE international, multi-centre, cross sectional survey

Introduction: Cardiopulmonary resuscitation (CPR) is often started irrespective of comorbidity or cause of arrest. We aimed to determine the prevalence of perception of inappropriate CPR of the last cardiac arrest encountered by clinicians working in emergency departments and out-of-hospital, factors associated with perception, and its relation to patient outcome. Methods: A cross-sectional survey was conducted in 288 centres in 24 countries. Factors associated with perception of CPR and outcome were analyzed by Cochran-Mantel-Haenszel tests and conditional logistic models. Results: Of the 4018 participating clinicians, 3150 (78.4%) perceived their last CPR attempt as appropriate, 548 (13.6%) were uncertain about its appropriateness and 320 (8.0%) perceived inappropriateness; survival to hospital discharge was 370/2412 (15.3%), 8/481 (1.7%) and 8/294 (2.7%) respectively. After adjusting for country, team and clinician's characteristics, the prevalence of perception of inappropriate CPR was higher for a non-shockable initial rhythm (OR 3.76 [2.13-6.64]; P 79 years) and in case of a "poor" first physical impression of the patient (3.45 [2.36-5.05]; P 79 years) and a "poor" first physical impression (0.26 [0.19-0.35]; P <0.0001). Conclusions: The perception of inappropriate CPR increased when objective indicators of poor prognosis were present and was associated with a low survival to hospital discharge. Factoring clinical judgment into the decision to (not) attempt CPR may reduce harm inflicted by excessive resuscitation attempts.Peer reviewe

Repositioning of the global epicentre of non-optimal cholesterol

High blood cholesterol is typically considered a feature of wealthy western countries(1,2). However, dietary and behavioural determinants of blood cholesterol are changing rapidly throughout the world(3) and countries are using lipid-lowering medications at varying rates. These changes can have distinct effects on the levels of high-density lipoprotein (HDL) cholesterol and non-HDL cholesterol, which have different effects on human health(4,5). However, the trends of HDL and non-HDL cholesterol levels over time have not been previously reported in a global analysis. Here we pooled 1,127 population-based studies that measured blood lipids in 102.6 million individuals aged 18 years and older to estimate trends from 1980 to 2018 in mean total, non-HDL and HDL cholesterol levels for 200 countries. Globally, there was little change in total or non-HDL cholesterol from 1980 to 2018. This was a net effect of increases in low- and middle-income countries, especially in east and southeast Asia, and decreases in high-income western countries, especially those in northwestern Europe, and in central and eastern Europe. As a result, countries with the highest level of non-HDL cholesterol-which is a marker of cardiovascular riskchanged from those in western Europe such as Belgium, Finland, Greenland, Iceland, Norway, Sweden, Switzerland and Malta in 1980 to those in Asia and the Pacific, such as Tokelau, Malaysia, The Philippines and Thailand. In 2017, high non-HDL cholesterol was responsible for an estimated 3.9 million (95% credible interval 3.7 million-4.2 million) worldwide deaths, half of which occurred in east, southeast and south Asia. The global repositioning of lipid-related risk, with non-optimal cholesterol shifting from a distinct feature of high-income countries in northwestern Europe, north America and Australasia to one that affects countries in east and southeast Asia and Oceania should motivate the use of population-based policies and personal interventions to improve nutrition and enhance access to treatment throughout the world.Peer reviewe

Rising rural body-mass index is the main driver of the global obesity epidemic in adults

Body-mass index (BMI) has increased steadily in most countries in parallel with a rise in the proportion of the population who live in cities(.)(1,2) This has led to a widely reported view that urbanization is one of the most important drivers of the global rise in obesity(3-6). Here we use 2,009 population-based studies, with measurements of height and weight in more than 112 million adults, to report national, regional and global trends in mean BMI segregated by place of residence (a rural or urban area) from 1985 to 2017. We show that, contrary to the dominant paradigm, more than 55% of the global rise in mean BMI from 1985 to 2017-and more than 80% in some low- and middle-income regions-was due to increases in BMI in rural areas. This large contribution stems from the fact that, with the exception of women in sub-Saharan Africa, BMI is increasing at the same rate or faster in rural areas than in cities in low- and middle-income regions. These trends have in turn resulted in a closing-and in some countries reversal-of the gap in BMI between urban and rural areas in low- and middle-income countries, especially for women. In high-income and industrialized countries, we noted a persistently higher rural BMI, especially for women. There is an urgent need for an integrated approach to rural nutrition that enhances financial and physical access to healthy foods, to avoid replacing the rural undernutrition disadvantage in poor countries with a more general malnutrition disadvantage that entails excessive consumption of low-quality calories.Peer reviewe