Clustered Cardiometabolic Risk, Cardiorespiratory Fitness and Physical Activity in 10-11 Year-Old Children. The CHANGE! Project Baseline

Objectives: The primary objective of this cross sectional pilot study was to report clustered risk scores combining traditional invasive with non invasive cardiometabolic risk markers in 10-11 year old children participating in the CHANGE! project at baseline. A secondary objective was to determine the relationship between clustered risk score and objectively measured physical activity (PA) and cardiorespiratory fitness (CRF). Methods: Habitual PA was measured using accelerometry and CRF (VO2peak) was assessed using an individually calibrated treadmill based protocol. Twenty-nine participants had valid data for all components of the clustered risk score, calculated using total cholesterol: high density lipoprotein-cholesterol (TC:HDL-C), glucose, systolic blood pressure (BP), LV Mass Index (g/m2.7), and trunk fat mass (g). Participants with a clustered risk score greater than 1SD above the mean, were categorised as ‘higher’ risk (n=6); all others were categorised as ‘normal’ risk. \ud \ud Results: Clustered risk score, controlling for somatic maturity and gender, was negatively correlated with VPA (r= -0.51, p=0.01), MVPA (r= -0.44, p=0.03) and VO2peak (r= -0.57, p<0.01). ANCOVA, with somatic maturity and gender as covariates, revealed that those in the ‘normal’ risk group were more fit than those in the ‘higher’ risk group [f (1,24)=4.518, p=0.044]). There were no statistically significant differences between risk groups and PA however mean data suggest that those in the ‘normal’ risk group accrued 4 minutes more daily VPA than the ‘higher’ risk group which may be clinically important. \ud \ud Conclusions: This provides further evidence of the importance of promoting CRF and VPA in children, to reduce cardiometabolic risk especially for those that are ‘higher’ risk

Cardiorespiratory fitness predicts clustered cardiometabolic risk in 10-11.9 year olds

The aim of this study was to investigate levels of clustered cardiometabolic risk and the odds of being ‘at risk’ according to cardiorespiratory fitness status in children. Data from 88 10–11.9-year-old children (mean age 11.05 ± 0.51 years), who participated in either the REACH Year 6 or the Benefits of Fitness Circuits for Primary School Populations studies were combined. Waist circumference, systolic blood pressure, diastolic blood pressure, glucose, triglycerides, high-density lipoprotein cholesterol, adiponectin and C-reactive protein were assessed and used to estimate clustered cardiometabolic risk. Participants were classified as ‘fit’ or ‘unfit’ using recently published definitions (46.6 and 41.9 mL/kg/min for boys and girls, respectively), and continuous clustered risk scores between fitness groups were assessed. Participants were subsequently assigned to a ‘normal’ or ‘high’ clustered cardiometabolic risk group based on risk scores, and logistic regression analysis assessed the odds of belonging to the increased cardiometabolic risk group according to fitness. The unfit group exhibited significantly higher clustered cardiometabolic risk scores (p < 0.001) than the fit group. A clear association between fitness group and being at increased cardiometabolic risk (B = 2.509, p = 0.001) was also identified, and participants classed as being unfit were found to have odds of being classified as ‘at risk’ of 12.30 (95 % CI = 2.64–57.33).\ud \ud Conclusion Assessing cardiorespiratory fitness is a valid method of identifying children most at risk of cardiometabolic pathologies. The ROC thresholds could be used to identify populations of children most at risk and may therefore be used to effectively target a cardiometabolic risk-reducing public health intervention

Clinical phenotype and outcome of hepatitis E virus - associated neuralgic amyotrophy

Objective: To determine the clinical phenotype and outcome in hepatitis E virus–associated neuralgic amyotrophy (HEV-NA). Methods: Cases of NA were identified in 11 centers from 7 European countries, with retrospective analysis of demographics, clinical/laboratory findings, and treatment and outcome. Cases of HEV-NA were compared with NA cases without evidence of HEV infection. Results: Fifty-seven cases of HEV-NA and 61 NA cases without HEV were studied. Fifty-six of 57 HEV-NA cases were anti-HEV IgM positive; 53/57 were IgG positive. In 38 cases, HEV RNA was recovered from the serum and in 1 from the CSF (all genotype 3). Fifty-one of 57 HEV-NA cases were anicteric; median alanine aminotransferase 259 IU/L (range 12–2,961 IU/L); in 6 cases, liver function tests were normal. HEV-NA cases were more likely to have bilateral involvement (80.0% vs 8.6%, p < 0.001), damage outside the brachial plexus (58.5% vs 10.5%, p < 0.01), including phrenic nerve and lumbosacral plexus injury (25.0% vs 3.5%, p = 0.01, and 26.4% vs 7.0%, p = 0.001), reduced reflexes (p = 0.03), sensory symptoms (p = 0.04) with more extensive damage to the brachial plexus. There was no difference in outcome between the 2 groups at 12 months. Conclusions: Patients with HEV-NA are usually anicteric and have a distinct clinical phenotype, with predominately bilateral asymmetrical involvement of, and more extensive damage to, the brachial plexus. Involvement outside the brachial plexus is more common in HEV-NA. The relationship between HEV and NA is likely to be causal, but is easily overlooked. Patients presenting with NA should be tested for HEV, irrespective of liver function test results. Prospective treatment/outcome studies of HEV-NA are warranted

ADDovenom: Thermostable Protein-Based ADDomer Nanoparticles as New Therapeutics for Snakebite Envenoming

Snakebite envenoming can be a life-threatening medical emergency that requires prompt medical intervention to neutralise the effects of venom toxins. Each year up to 138,000 people die from snakebites and threefold more victims suffer life-altering disabilities. The current treatment of snakebite relies solely on antivenom—polyclonal antibodies isolated from the plasma of hyperimmunised animals—which is associated with numerous deficiencies. The ADDovenom project seeks to deliver a novel snakebite therapy, through the use of an innovative protein-based scaffold as a next-generation antivenom. The ADDomer is a megadalton-sized, thermostable synthetic nanoparticle derived from the adenovirus penton base protein; it has 60 high-avidity binding sites to neutralise venom toxins. Here, we outline our experimental strategies to achieve this goal using state-of-the-art protein engineering, expression technology and mass spectrometry, as well as in vitro and in vivo venom neutralisation assays. We anticipate that the approaches described here will produce antivenom with unparalleled efficacy, safety and affordability

LTR Retrotransposons Contribute to Genomic Gigantism in Plethodontid Salamanders

Among vertebrates, most of the largest genomes are found within the salamanders, a clade of amphibians that includes 613 species. Salamander genome sizes range from ∼14 to ∼120 Gb. Because genome size is correlated with nucleus and cell sizes, as well as other traits, morphological evolution in salamanders has been profoundly affected by genomic gigantism. However, the molecular mechanisms driving genomic expansion in this clade remain largely unknown. Here, we present the first comparative analysis of transposable element (TE) content in salamanders. Using high-throughput sequencing, we generated genomic shotgun data for six species from the Plethodontidae, the largest family of salamanders. We then developed a pipeline to mine TE sequences from shotgun data in taxa with limited genomic resources, such as salamanders. Our summaries of overall TE abundance and diversity for each species demonstrate that TEs make up a substantial portion of salamander genomes, and that all of the major known types of TEs are represented in salamanders. The most abundant TE superfamilies found in the genomes of our six focal species are similar, despite substantial variation in genome size. However, our results demonstrate a major difference between salamanders and other vertebrates: salamander genomes contain much larger amounts of long terminal repeat (LTR) retrotransposons, primarily Ty3/gypsy elements. Thus, the extreme increase in genome size that occurred in salamanders was likely accompanied by a shift in TE landscape. These results suggest that increased proliferation of LTR retrotransposons was a major molecular mechanism contributing to genomic expansion in salamanders

Cardiometabolic risk in 10 to 11 year old children : the impacts of physical activity, cardiorespiratory fitness, body composition and lifestyle education

The aim of this thesis was to investigate the impacts of physical activity (PA), cardiorespiratory fitness (CRF), body composition and lifestyle education on cardiometabolic (CM) risk in 10-11 year old children. This broad aim was approached using three studies. Studies 1 and 2 were cross sectional observational studies, and study 3 was a clustered randomised control trial, with intervention effects assessed at post intervention and again at 8 to 10 weeks after the intervention. Initially, in the first cross sectional study (Chapter 4) the relationships between non- invasive (LV Mass, E/A, E'/A', E/E', trunk fat mass, whole body fat mass) and invasive CM risk markers (CRP, HOMA-IR, adiponectin, TC: HDL-C), and between all risk markers and CRF (V02 peak), time spent sedentary, moderate to vigorous intensity PA (MVPA) and vigorous PA (VPA) were investigated in 10-11 year old children (n=62). The key findings were significant but generally weak relationships present between some of the non- invasive and invasive markers of CM risk and risk markers also had significant correlations with measures of CRF and PA. CRP was significantly positively correlated with whole body fat in boys (rho=0.486, p<0.05) and girls (rho = 0.485 , p<O.Ol) and with trunk fat mass in boys (rho = 0.384, p<0.05) and girls (rho =0.489, p<O.Ol). Adiponectin was negatively correlated with whole body fat (rho = -0.446, p<0.05, and R= -0.697, p<O.Ol) and trunk fat mass (rho = -0.614, p<O.Ol; rho = -0.475, p<O.Ol) in boys and girls respectively, and in girls adiponectin also correlated positively with E'/A' (r=0.356, p<0.05). In boys only, TC:HDL-C was positively correlated with whole body fat (rho =0.407, p<O.Ol) and trunk fat mass (rho =0.391, p<0.05). ; V02Peak was negatively correlated with CRP in boys (Rho = -0.492, p<0.05) and HOMA-IR in girls (Rho =-0.522, p<O.Ol). V02Peak was also negatively correlated with whole body fat (rho =-0.515, p<O.Ol; r=-0.697, p<O.Ol) and trunk fat mass (rho=-0.494, p<0.05; rho =-0.706, p<O.Ol) in boys and girls respectively. Both MVPA and VPA correlated negatively with TC: HDL-C in girls (rho= -0.396, p<0.05; rho =-0.428, p<0.05) and MVPA correlated with whole body fat (rho= -0.602, p<O.Ol) and trunk fat mass (rho=-0.65, p<O.Ol) in boys. VPA also correlated with whole body fat in girls (rho= - 0.544, p<O.Ol) and with trunk fat mass in both boys (rho= -0.428, p<0.05) and girls (rho= - 0.468, p<O.Ol). Time spent sedentary had a positive correlation with whole body fat in boys (rho = 0.429, p<0.05). This study demonstrated that risk factors clustered in individuals and that relationships were present between invasive and non-invasive markers of cardiometabolic risk, and provided preliminary evidence to investigate this phenomenon further. The correlations described in this study suggest a clustered risk score which includes both invasive and non-invasive measures may add value to predicting overall risk. The second cross sectional study (Chapter 5) investigated clustered CM risk, by combining invasive markers with non-invasive 'pre-clinical' markers of CM risk into a clustered risk score, in a different cohort of 10 - 11 year old children. Clustered risk scores were negatively correlated with CRF and PA. V02 peakshowed a moderate negative correlation with CRS A (r=-0.57, p<O.Ol) and CRS B (r= -0.60, p<O.Ol) VPA showed a moderate negative correlation with CRS A (r= -0.51, p= 0.01) and CRS B (r= -0.50, p=O.Ol). MVPA showed a moderate negative correlation with CRS A (r= -0.44, p= 0.03) and CRS B [r= -0.41, p=0.04). Sedentary time showed a moderate positive correlation with CRS A (r= 0.414, p= 0.049). The evidence provided by these two observational studies, Study 1 (Chapter 4) and Study 2 (Chapter 5), along with other literature, as discussed throughout this thesis, gave rationale for an intervention with the aim to reduce negative lifestyle behaviours, of low levels of PA, high levels of sedentary behaviour and poor nutritional balance, increase CRF and maintain a healthy body weight. The final study was a clustered randomised control trial which investigated the immediate and short term (8 to 10 weeks follow up) effects of the Children's Health Activity and Nutrition: Get Educated! (CHANGE!), curriculum based multi-disciplinary PA and nutrition intervention, on CM risk in 10 to 11 year old children. Whilst there were some statistically significant intervention effects on waist circumference (WC) (Adjusted mean (SE) change for Control = +1.4 cm (0.3) [95% Cl 0.7, 2.1]; Intervention = -0.1 cm (0.4) [95% Cl -0.9,0.6], p=0.006) and diastolic blood pressure (dBP) (control group adjusted mean (SE) = +3 (3) [95% Cl -4, 9] mmHg; intervention = -14 (3) 95% Cl [-21, -7] mmHg) at post intervention these were not sustained at 8 to 10 week follow up. There were however improvements demonstrated in CRF in the intervention group at follow up (Adjusted mean change for control = -2.8 ml/kg/min (1.5) [95% Cl -5.9, 0.3]; Intervention= +3.8 ml/kg/min (1.6) [95% Cl 0.5, 7.1], p=0.009). Overall the studies included in this thesis have highlighted a number of relationships between CM risk markers, measures of body composition, CRF and PA. Hence there is still a need to intervene in this population to reduce modifiable risk. The lifestyle education intervention (Study 3) demonstrated some success; however this was limited due to a number of factors, namely the lack of statistical power, intervention fidelity, and the lack of prescriptive PA element. Investment in promoting healthy lifestyles is essential in ensuring the future health of children as they develop into adulthood. There is a clear need for interventions which are sustainable, and lifestyle education embedded into the school curriculum is a logical and feasible option to reach the whole target population. Further research is required to evaluate the optimum sustainable intervention to improve children's CM health in the long term.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Cardiometabolic risk in 10 to 11 year old children : the impacts of physical activity, cardiorespiratory fitness, body composition and lifestyle education

The aim of this thesis was to investigate the impacts of physical activity (PA), cardiorespiratory fitness (CRF), body composition and lifestyle education on cardiometabolic (CM) risk in 10-11 year old children. This broad aim was approached using three studies. Studies 1 and 2 were cross sectional observational studies, and study 3 was a clustered randomised control trial, with intervention effects assessed at post intervention and again at 8 to 10 weeks after the intervention. Initially, in the first cross sectional study (Chapter 4) the relationships between non- invasive (LV Mass, E/A, E'/A', E/E', trunk fat mass, whole body fat mass) and invasive CM risk markers (CRP, HOMA-IR, adiponectin, TC: HDL-C), and between all risk markers and CRF (V02 peak), time spent sedentary, moderate to vigorous intensity PA (MVPA) and vigorous PA (VPA) were investigated in 10-11 year old children (n=62). The key findings were significant but generally weak relationships present between some of the non- invasive and invasive markers of CM risk and risk markers also had significant correlations with measures of CRF and PA. CRP was significantly positively correlated with whole body fat in boys (rho=0.486, p<0.05) and girls (rho = 0.485 , p<O.Ol) and with trunk fat mass in boys (rho = 0.384, p<0.05) and girls (rho =0.489, p<O.Ol). Adiponectin was negatively correlated with whole body fat (rho = -0.446, p<0.05, and R= -0.697, p<O.Ol) and trunk fat mass (rho = -0.614, p<O.Ol; rho = -0.475, p<O.Ol) in boys and girls respectively, and in girls adiponectin also correlated positively with E'/A' (r=0.356, p<0.05). In boys only, TC:HDL-C was positively correlated with whole body fat (rho =0.407, p<O.Ol) and trunk fat mass (rho =0.391, p<0.05). ; V02Peak was negatively correlated with CRP in boys (Rho = -0.492, p<0.05) and HOMA-IR in girls (Rho =-0.522, p<O.Ol). V02Peak was also negatively correlated with whole body fat (rho =-0.515, p<O.Ol; r=-0.697, p<O.Ol) and trunk fat mass (rho=-0.494, p<0.05; rho =-0.706, p<O.Ol) in boys and girls respectively. Both MVPA and VPA correlated negatively with TC: HDL-C in girls (rho= -0.396, p<0.05; rho =-0.428, p<0.05) and MVPA correlated with whole body fat (rho= -0.602, p<O.Ol) and trunk fat mass (rho=-0.65, p<O.Ol) in boys. VPA also correlated with whole body fat in girls (rho= - 0.544, p<O.Ol) and with trunk fat mass in both boys (rho= -0.428, p<0.05) and girls (rho= - 0.468, p<O.Ol). Time spent sedentary had a positive correlation with whole body fat in boys (rho = 0.429, p<0.05). This study demonstrated that risk factors clustered in individuals and that relationships were present between invasive and non-invasive markers of cardiometabolic risk, and provided preliminary evidence to investigate this phenomenon further. The correlations described in this study suggest a clustered risk score which includes both invasive and non-invasive measures may add value to predicting overall risk. The second cross sectional study (Chapter 5) investigated clustered CM risk, by combining invasive markers with non-invasive 'pre-clinical' markers of CM risk into a clustered risk score, in a different cohort of 10 - 11 year old children. Clustered risk scores were negatively correlated with CRF and PA. V02 peakshowed a moderate negative correlation with CRS A (r=-0.57, p<O.Ol) and CRS B (r= -0.60, p<O.Ol) VPA showed a moderate negative correlation with CRS A (r= -0.51, p= 0.01) and CRS B (r= -0.50, p=O.Ol). MVPA showed a moderate negative correlation with CRS A (r= -0.44, p= 0.03) and CRS B [r= -0.41, p=0.04). Sedentary time showed a moderate positive correlation with CRS A (r= 0.414, p= 0.049). The evidence provided by these two observational studies, Study 1 (Chapter 4) and Study 2 (Chapter 5), along with other literature, as discussed throughout this thesis, gave rationale for an intervention with the aim to reduce negative lifestyle behaviours, of low levels of PA, high levels of sedentary behaviour and poor nutritional balance, increase CRF and maintain a healthy body weight. The final study was a clustered randomised control trial which investigated the immediate and short term (8 to 10 weeks follow up) effects of the Children's Health Activity and Nutrition: Get Educated! (CHANGE!), curriculum based multi-disciplinary PA and nutrition intervention, on CM risk in 10 to 11 year old children. Whilst there were some statistically significant intervention effects on waist circumference (WC) (Adjusted mean (SE) change for Control = +1.4 cm (0.3) [95% Cl 0.7, 2.1]; Intervention = -0.1 cm (0.4) [95% Cl -0.9,0.6], p=0.006) and diastolic blood pressure (dBP) (control group adjusted mean (SE) = +3 (3) [95% Cl -4, 9] mmHg; intervention = -14 (3) 95% Cl [-21, -7] mmHg) at post intervention these were not sustained at 8 to 10 week follow up. There were however improvements demonstrated in CRF in the intervention group at follow up (Adjusted mean change for control = -2.8 ml/kg/min (1.5) [95% Cl -5.9, 0.3]; Intervention= +3.8 ml/kg/min (1.6) [95% Cl 0.5, 7.1], p=0.009). Overall the studies included in this thesis have highlighted a number of relationships between CM risk markers, measures of body composition, CRF and PA. Hence there is still a need to intervene in this population to reduce modifiable risk. The lifestyle education intervention (Study 3) demonstrated some success; however this was limited due to a number of factors, namely the lack of statistical power, intervention fidelity, and the lack of prescriptive PA element. Investment in promoting healthy lifestyles is essential in ensuring the future health of children as they develop into adulthood. There is a clear need for interventions which are sustainable, and lifestyle education embedded into the school curriculum is a logical and feasible option to reach the whole target population. Further research is required to evaluate the optimum sustainable intervention to improve children's CM health in the long term.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Scaling of Peak Oxygen Uptake in Children:A Comparison of Three Body Size Index Models

Purpose: We aimed to compare three candidate body size index models for the scaling of aerobic fitness (V˙ O2peak) in children: whole body mass, total lean body mass, and the lean mass of both legs. Methods: V˙ O2peak and total lean mass of the body and both legs (via dual-energy x-ray absorptiometry) were assessed in 126 girls and 87 boys aged 9–11 yr. We applied nonlinear allometric models of the formV˙ O2peak = a�body sizeb, adjusted for biological sex and maturity offset (years from peak height velocity). We assessed goodness of fit using the Akaike information criterion. Results: The Akaike weights (Akaike differences) were as follows: lean mass of both legs = 0.69 (0), total lean body mass = 0.31 (1.6), and whole body mass = G1ej8 (36.6). The size exponent(90% confidence interval) for the lean mass of both legs was 0.55 (0.46–0.64). V˙ O2peak was 17% (13%–21%) lower in girls after controlling for the lean mass of both legs and maturity offset. After controlling for body size and sex, a 1-yr increase in maturity offset (closer to peak height velocity) was associated with a 6% (4%–9%) higher V˙ O2peak. Conclusions: Allometric scaling of V˙ O2peak by the lean mass of both legs provides the best model for quantifying growth-related changes in aerobic fitness in pediatric populations, although this model is only marginally superior to the total lean body mass model. There is no support for the total body mass model. Maturity and sex are also important covariates exerting a size-independent influence on peak aerobic fitness

ADDovenom : Thermostable Protein-Based ADDomer Nanoparticles as New Therapeutics for Snakebite Envenoming

Snakebite envenoming can be a life-threatening medical emergency that requires prompt medical intervention to neutralise the effects of venom toxins. Each year up to 138,000 people die from snakebites and threefold more victims suffer life-altering disabilities. The current treatment of snakebite relies solely on antivenom—polyclonal antibodies isolated from the plasma of hyperimmunised animals—which is associated with numerous deficiencies. The ADDovenom project seeks to deliver a novel snakebite therapy, through the use of an innovative protein-based scaffold as a next-generation antivenom. The ADDomer is a megadalton-sized, thermostable synthetic nanoparticle derived from the adenovirus penton base protein; it has 60 high-avidity binding sites to neutralise venom toxins. Here, we outline our experimental strategies to achieve this goal using state-of-the-art protein engineering, expression technology and mass spectrometry, as well as in vitro and in vivo venom neutralisation assays. We anticipate that the approaches described here will produce antivenom with unparalleled efficacy, safety and affordability