The impact of school exclusion in childhood on health and well-being outcomes in adulthood: Estimating causal effects using inverse probability of treatment weighting.

BACKGROUND: Previous evidence has suggested a strong association between school exclusion and health outcomes. However, as health risks are themselves related to the risk of experiencing a school exclusion, it has been challenging to determine the extent to which school exclusion impacts later health outcomes, as opposed to reflecting a marker for pre-existing risks. AIM: The aim of the current study was to address this challenge in estimating the medium-to-long-term impact of school exclusion of health and well-being outcomes. METHODS: To this end, we used an inverse propensity weighting approach in the Next Steps data set (N = 6534, from wave 1, 2014, to wave 8, 2015). RESULTS: We found that after weighting for propensity of treatment scores estimated based on a wide range of factors, including previous health indicators, there was a significant effect of school exclusion on a wide range of health and well-being outcomes. DISCUSSION: These results provide some of the most robust evidence to date that school exclusion harms long-term health outcomes. CONCLUSION: The findings suggest that policies should aim to reduce exclusion and ensure access to preventative health support for those who experience a school exclusion.

Long-term labour market and economic consequences of school exclusions in England: Evidence from two counterfactual approaches.

BACKGROUND: Previous research suggests that school exclusion during childhood is a precursor to social exclusion in adulthood. Past literature on the consequences of school exclusion is, however, scarce and mainly focused on short-term outcomes such as educational attainment, delinquency, and mental health in early adolescence. Moreover, this evidence is based primarily on descriptive and correlational analysis, whereas robust causal evidence is required to best inform policy. AIMS: We aimed to estimate the mid-to-long-term impact of school exclusion on labour market and economic outcomes. SAMPLE: The sample included 6,632 young people who at the age of 25/26 in the year 2015 participated in the Next Steps survey of whom 86 were expelled from school and 711 were suspended between the ages of 13/14 and 16/17. METHOD: Using high quality existing longitudinal data, we utilized four approaches to evaluate the impact of school exclusion: logistic regression-adjustment models, propensity score matching, school fixed-effects analysis, and inverse propensity weighting. The latter two counterfactual approaches were used to estimate causal effects. RESULTS: We found that school exclusion increased the risk of becoming NEET at the age of 19/20, and then remaining economically inactive at the age of 25/26, as well as experiencing higher unemployment risk and earning lower wages also at the age of 25/26. CONCLUSION: School exclusion has pervasive negative effects into adulthood. Policy interventions should focus on both prevention and mitigating its negative effects. Interventions aimed at re-integrating excluded individuals into education or vocational training could be key in reducing the risk of poor socio-economic outcomes and social exclusion.

Vitrification as an Alternative Approach for Sperm Cryopreservation in Marine Fishes.

The Southern Flounder Paralichthys lethostigma is a high-value species and a promising aquaculture candidate. Because sperm volume can be limited in this species (<500 µL), new sperm cryopreservation methods need to be evaluated. Vitrification is an alternative to conventional slow-rate freezing, whereby small volumes are cryopreserved at high cooling rates (>1,000°C/min). The goal of this work was to develop a standardized approach for vitrification of Southern Flounder sperm. The specific objectives were to (1) evaluate thawing methods and vitrification solutions, (2) evaluate the postthaw membrane integrity of sperm vitrified in different cryoprotectant solutions, (3) examine the relationship between membrane integrity and motility, and (4) evaluate the ability of vitrified sperm to fertilize eggs. From the vitrification solutions tested, the highest postthaw motility (28 ± 9% [mean ± SD]) and membrane integrity (11 ± 4%) was observed for 20% ethylene glycol plus 20% glycerol. There was no significant difference in postthaw motility of sperm thawed at 21°C or at 37°C. Fertilization from vitrified sperm in one trial yielded the same fertilization rate (50 ± 20%) as the fresh sperm control, while the sperm from the other two males yielded 3%. This is the first report of fertilization by vitrified sperm in a marine fish. Vitrification can be simple, fast, inexpensive, performed in the field, and, at least for small fishes, offers an alternative to conventional cryopreservation. Because of the minute volumes needed for ultrarapid cooling, vitrification is not presently suited as a production method for large fishes. Vitrification can be used to reconstitute lines from valuable culture species and biomedical models, conserve mutants for development of novel lines for ornamental aquaculture, and transport frozen sperm from the field to the repository to expand genetic resources.

Coupling aquaculture with forest plantations for food, energy, and water resiliency.

Freshwater aquaculture and forest bioenergy markets are expanding globally in areas concurrently experiencing human population growth, urbanization and water shortages. Coupling these agroecosystems can improve food, energy, and water resiliency by enhancing ecosystem services through fertilization, water-reuse, carbon storage, and bioenergy via biomass production. This study evaluated how a model aquaculture-managed forest plantation could (1) provision fish and woody biomass; (2) regulate carbon, groundwater infiltration, and groundwater quality; and (3) support nutrient cycling over a two-year period. A 0.5-hectare hardwood bioenergy plantation was established with 12 Populus spp. genotypes adjacent to a 0.6-hectare freshwater aquaculture operation (hybrid striped bass, Morone chrysops×M. saxatilis); pond waters were land-applied on the plantation for two years. The aquaculture operation produced ~3.5Mg of fish and trees yielded 5.9Mgha(-1)yr(-1) of oven-dry biomass, sequestered 2.9Mg carbon (C) ha(-1)yr(-1) and stored 0.028Mg nitrogen (N) ha(-1)yr(-1). Biomass productivity, carbon storage, and nitrogen storage differed significantly among the evaluated Populus genotypes. Land application of pond water increased groundwater infiltration by 60% relative to the previous year. The integrated system regulated chlorophyll a, total organic carbon, and nitrogen in groundwater at concentrations below regulatory limits. This study demonstrated that coupled agroecosystems could deliver productive yields of food and bioenergy as well as support water re-use while meeting water quality regulations. More research is needed to evaluated long-term sustainability and economic viability of this coupled system and other land management practices that seek to improve food, energy, and water resiliency.

Mechanisms of Egg Yolk Formation and Implications on Early Life History of White Perch (Morone americana).

The three white perch (Morone americana) vitellogenins (VtgAa, VtgAb, VtgC) were quantified accurately and precisely in the liver, plasma, and ovary during pre-, early-, mid-, and post-vitellogenic oocyte growth using protein cleavage-isotope dilution mass spectrometry (PC-IDMS). Western blotting generally mirrored the PC-IDMS results. By PC-IDMS, VtgC was quantifiable in pre-vitellogenic ovary tissues and VtgAb was quantifiable in pre-vitellogenic liver tissues however, neither protein was detected by western blotting in these respective tissues at this time point. Immunohistochemistry indicated that VtgC was present within pre-vitellogenic oocytes and localized to lipid droplets within vitellogenic oocytes. Affinity purification coupled to tandem mass spectrometry using highly purified VtgC as a bait protein revealed a single specific interacting protein (Y-box binding protein 2a-like [Ybx2a-like]) that eluted with suramin buffer and confirmed that VtgC does not bind the ovary vitellogenin receptors (LR8 and Lrp13). Western blotting for LR8 and Lrp13 showed that both receptors were expressed during vitellogenesis with LR8 and Lrp13 expression highest in early- and mid-vitellogenesis, respectively. The VtgAa within the ovary peaked during post-vitellogenesis, while VtgAb peaked during early-vitellogenesis in both white perch and the closely related striped bass (M. saxatilis). The VtgC was steadily accumulated by oocytes beginning during pre-vitellogenesis and continued until post-vitellogenesis and its composition varies widely between striped bass and white perch. In striped bass, the VtgC accounted for 26% of the vitellogenin-derived egg yolk, however in the white perch it comprised only 4%. Striped bass larvae have an extended developmental window and these larvae have yolk stores that may enable them to survive in the absence of food for twice as long as white perch after hatch. Thus, the VtgC may play an integral role in providing nutrients to late stage fish larvae prior to the onset of exogenous feeding and its composition in the egg yolk may relate to different early life histories among this diverse group of animals.

Machine learning reveals sex-specific 17ß-estradiol-responsive expression patterns in white perch (Morone americana) plasma proteins.

With growing abundance and awareness of endocrine disrupting compounds (EDCs) in the environment, there is a need for accurate and reliable detection of EDC exposure. Our objective in the present study was to observe differences within and between the global plasma proteomes of sexually mature male and female white perch (Morone americana) before (Initial Control, IC) and after 17ß-estradiol (E2 ) induction. Semiquantitative nanoLC-MS/MS data were analyzed by machine learning support vector machines (SVMs) and by two-way ANOVA. By ANOVA, the expression levels of 44, 77, and 57 proteins varied significantly by gender, treatment, and the interaction of gender and treatment, respectively. SVMs perfectly classified male and female perch IC and E2 -induced plasma samples using the protein expression data. E2 -induced male and female perch plasma proteomes contained significantly higher levels of the yolk precursors vitellogenin Aa and Ab (VtgAa, VtgAb), as well as latrophilin and seven transmembrane domain-containing protein 1 (Eltd1) and kininogen 1 (Kng1). This is the first report that Eltd1 and Kng1 may be E2 -responsive proteins in fishes and therefore may be useful indicators of estrogen induction.

The prevention of anxiety in children through school-based interventions: study protocol for a 24-month follow-up of the PACES project.

BACKGROUND: Anxiety in children is common and incapacitating and increases the risk of mental health disorders in adulthood. Although effective interventions are available, few children are identified and referred for specialist treatment. Alternative approaches in which prevention programmes are delivered in school appear promising. However, comparatively little is known about the best intervention leader (health care-led vs. school-led), long-term effects or the primary preventive value of such programmes. METHODS/DESIGN: Preventing Anxiety in Children through Education in Schools, or PACES, is a pragmatic cluster randomised controlled trial evaluating the effectiveness of a cognitive-behavioural therapy prevention programme (FRIENDS) on symptoms of anxiety and low mood in 9- to 10-year-old children. Forty-one schools were randomly assigned to one of three conditions: school-led FRIENDS, health care-led FRIENDS or treatment as usual. Assessments were undertaken at baseline, 6 months and 12 months, with the primary outcome measure being the Revised Child Anxiety and Depression Scale score at 12 months. Secondary outcome measures are changes in self-esteem, worries, bullying and life satisfaction. DISCUSSION: This protocol summarises the procedure for the 24-month follow-up of this cohort. The study will determine the medium-term effectiveness of an anxiety prevention programme delivered in schools. TRIAL REGISTRATION: ISRCTN23563048.

Compartment proteomics analysis of white perch (Morone americana) ovary using support vector machines.

Compartment proteomics enable broad characterization of target tissues. We employed a simple fractionation method and filter-aided sample preparation (FASP) to characterize the cytosolic and membrane fractions of white perch ovary tissues by semiquantitative tandem mass spectrometry using label-free quantitation based on normalized spectral counts. FASP depletes both low-molecular-weight and high-molecular-weight substances that could interfere with protein digestion and subsequent peptide separation and detection. Membrane proteins are notoriously difficult to characterize due to their amphipathic nature and association with lipids. The simple fractionation we employed effectively revealed an abundance of proteins from mitochondria and other membrane-bounded organelles. We further demonstrate that support vector machines (SVMs) offer categorical classification of proteomics data superior to that of parametric statistical methods such as analysis of variance (ANOVA). Specifically, SVMs were able to perfectly (100% correct) classify samples as either membrane or cytosolic fraction during cross-validation based on the expression of 242 proteins with the highest ANOVA p-values (i.e., those that were not significant for enrichment in either fraction). The white perch ovary cytosolic and membrane proteomes and transcriptome presented in this study can support future investigations into oogenesis and early embryogenesis of white perch and other members of the genus Morone.

Classroom-based cognitive behaviour therapy (FRIENDS): a cluster randomised controlled trial to Prevent Anxiety in Children through Education in Schools (PACES).

BACKGROUND: Anxiety in children is common, impairs everyday functioning, and increases the risk of severe mental health disorders in adulthood. We investigated the effect of a classroom-based cognitive behaviour therapy prevention programme (FRIENDS) on anxiety symptoms in children. METHODS: Preventing Anxiety in Children though Education in Schools (PACES) is a three-group parallel cluster randomised controlled trial. Interventions were given between September, 2011, and July, 2012, with schools as the unit of allocation and individual participants as the unit of analysis. We enrolled state-funded junior schools in southwest England. We sent information to all eligible schools (state-funded junior schools in southwest England) inviting them to enrol in the study. School year groups were assigned by computer-generated randomisation (1:1:1) to receive either school-led FRIENDS (led by teacher or school staff member), health-led FRIENDS (led by two trained health facilitators), or usual school provision. Children were not masked to treatment allocation. The allocated programme was given to all students (aged 9-10 years) in the school year (ie, universal delivery) as part of the school curriculum as nine, 60 min weekly sessions. Outcomes were collected by self-completed questionnaire administered by researchers masked to allocation. Primary outcome was symptoms of anxiety and low mood at 12 months assessed by the Revised Child Anxiety and Depression Scale (RCADS 30). Analyses were intention to treat and accounted for the clustered nature of the design. The study is registered, number ISRCTN23563048. FINDINGS: 45 schools were enrolled: 14 (n=497 children) were randomly assigned to school-led FRIENDS, 14 (n=509) to health-led FRIENDS, and 12 (n=442) to usual school provision. 1257 (92%) children completed 12 month assessments (449 in health-led FRIENDS, 436 in school-led FRIENDS, and 372 in usual school provision). We recorded a difference at 12 months in adjusted mean child-reported RCADS scores for health-led versus school-led FRIENDS (19·49 [SD 14·81] vs 22·86 [15·24]; adjusted difference -3·91, 95% CI -6·48 to -1·35; p=0·0004) and health-led FRIENDS versus usual school provision (19·49 [14·81] vs 22·48 [15·74]; -2·66, -5·22 to -0·09; p=0·043). We noted no differences in parent or teacher ratings. Training teachers to deliver mental health programmes was not as effective as delivery by health professionals. INTERPRETATION: Universally delivered anxiety prevention programmes can be effective when used in schools. However, programme effectiveness varies depending on who delivers them. FUNDING: National Institute for Health Research Public Health Research Programme.

Masculinizing effect of background color and cortisol in a flatfish with environmental sex-determination.

Environmental sex-determination (ESD) is the phenomenon by which environmental factors regulate sex-determination, typically occurring during a critical period of early development. Southern flounder (Paralichthys lethostigma) exhibit temperature-dependent sex-determination that appears to be restricted to the presumed XX female genotype with the extremes of temperature, both high and low, skewing sex ratios toward males. In order to evaluate other environmental factors that may influence sex-determination, we investigated the influence of background color and cortisol on sex-determination in southern flounder. Experiments involving three sets of tanks, each painted a different color, were conducted at different temperatures using southern flounder of mixed XX-XY genotype. The studies involved rearing juvenile southern flounder in either black, gray, or blue tanks and sex-determination was assessed by gonadal histology. In both studies, blue tanks showed significant male-biased sex ratios (95 and 75% male) compared with black and gray tanks. The stress corticosteroid cortisol may mediate sex-determining processes associated with environmental variables. Cortisol from the whole body was measured throughout the second experiment and fishes in blue tanks had higher levels of cortisol during the period of sex-determination. These data suggest that background color can be a cue for ESD, with blue acting as a stressor during the period of sex-determination, and ultimately producing male-skewed populations. In a separate study using XX populations of southern flounder, cortisol was applied at 0, 100, or 300 mg/kg of gelatin-coated feed. Fish were fed intermittently prior to, and just through, the period of sex-determination. Levels of gonadal P450 aromatase (cyp19a1) and forkhead transcription factor L2 (FoxL2) messenger RNA (mRNA) were measured by qRT-PCR as markers for differentiation into females. Müllerian-inhibiting substance mRNA was used as a marker of males' gonadal development. Control fish showed female-biased sex ratios approaching 100%, whereas treatment with 100 mg/kg cortisol produced 28.57% females and treatment with 300 mg/kg cortisol produced only 13.33% females. These results suggest that cortisol is a critical mediator of sex-determination in southern flounder by promoting masculinization. This linkage between the endocrine stress axis and conserved sex-determination pathways may provide a mechanism for adaptive modification of sex ratio in a spatially and temporally variable environment.

School-based intervention to reduce anxiety in children: study protocol for a randomized controlled trial (PACES).

BACKGROUND: Emotional problems such as anxiety and low mood in children are common, impair everyday functioning and increase the risk of severe mental health disorders in adulthood. Relatively few children with emotional health problems are identified and referred for treatment indicating the need to investigate preventive approaches. METHODS/DESIGN: The study is designed to be a pragmatic cluster randomized controlled trial evaluating the effectiveness of an efficacious school-based cognitive behavior therapy (CBT) prevention program (FRIENDS) on symptoms of anxiety and low mood in children 9 to 10 years of age. The unit of allocation is schools which are assigned to one of three conditions: school-led FRIENDS, health-led FRIENDS or treatment as usual. Assessments will be undertaken at baseline, 6 months and 12 months. The primary outcome measure is change on the Revised Child Anxiety and Depression Scale. Secondary outcome measures assess changes in self-esteem, worries, bullying and life satisfaction. An economic evaluation will be undertaken. DISCUSSION: As of September 2011, 41 schools have been recruited and randomized. Final 12-month assessments are scheduled to be completed by May 2013. TRIAL REGISTRATION: ISRCTN23563048.

Sex determination in flatfishes: Mechanisms and environmental influences.

Flounder of the genus Paralichthys exhibit a unique mode of sex determination where both low and high temperatures induce male-skewed sex ratios, while intermediate temperatures produce a 1:1 sex ratio. Male differentiation is thus easily induced in genetic females creating a combination of genetic (GSD) and environmental sex determination (ESD). Since male flounder become reproductively fit at substantially smaller body sizes than females, temperature or other environmental variables that elicit lower growth rates may also influence sex differentiation toward male development. This review covers our current knowledge of sex determination and differentiation in flatfishes including possible adaptive significance of ESD and involvement of factors such as aromatase (cyp19).

Temperature affects insulin-like growth factor I and growth of juvenile southern flounder, Paralichthys lethostigma.

Temperature profoundly influences growth of heterothermic vertebrates. However, few studies have investigated the effects of temperature on growth and insulin-like growth factor I (IGF-I) in fishes. The aim of this study was to examine effects of temperature on growth and establish whether IGF-I may mediate growth at different temperatures in southern flounder, Paralichthys lethostigma. In two experiments, juvenile flounder were reared at 23 and 28 degrees C and growth was monitored for either 117 or 197 days. Growth was similar across treatments in both experiments until fish reached approximately 100 mm total length. Body size then diverged with fish at 23 degrees C ultimately growing 65-83% larger than those at 28 degrees C. Muscle IGF-I mRNA, plasma IGF-I, and hepatosomatic index (HSI) were significantly higher in flounder at 23 degrees C, whereas hepatic IGF-I mRNA abundance did not differ with treatment. Muscle IGF-I mRNA was correlated with HSI, while plasma IGF-I was correlated with body size, hepatic IGF-I mRNA, and HSI. These results demonstrate a strong effect of temperature on flounder growth and show that temperature-induced variation in growth is associated with differences in systemic IGF-I and local (i.e., muscle) IGF-I mRNA levels. The results also support the use of plasma IGF-I and HSI as indicators of flounder growth status.

Aromatase cytochrome P450: cloning, intron variation, and ontogeny of gene expression in southern flounder (Paralichthys lethostigma).

Aromatase cytochrome P450 (P450arom) is the enzyme complex responsible for conversion of androgens to estrogens in vertebrates. Consequently, in some fishes its activity appears critical to ovarian differentiation. Southern flounder (Paralichthys lethostigma) is a commercially important flatfish in which females grow larger than males and sex determination is temperature sensitive. Through cloning of the P450arom gene in ovary and quantitative reverse transcription-polymerase chain reaction, we developed a biomarker for early female differentiation in southern flounder. The deduced amino acid sequence for southern flounder P450arom is similar to other teleosts. Comparison of P450arom intron sequences from fish of different populations revealed substantial inter-individual variation. Adult ovary and spleen exhibited high levels of P450arom mRNA, while P450arom mRNA was only weakly detected in testes. Brain, liver, intestine, kidney, gill, muscle, and heart showed little or no P450arom mRNA expression. Gonads of wild and hatchery-produced juvenile flounder of sizes spanning the period of sex differentiation initially exhibited low levels of P450arom mRNA followed by increases in some individuals and bifurcation into two clearly segregated groups (i.e., putative males and females) beginning at approximately 65 mm in total length. Gonadal histology confirmed predictions of sex based on P450arom expression in juvenile flounder, demonstrating that the patterns of P450arom expression observed relate to sex-specific differentiation. This research represents a unique approach to assessing sex differentiation in a natural population, and a powerful technique for better understanding mechanisms of flounder sex determination and rapidly defining conditions for controlling sex for aquaculture.

The influence of school culture on smoking among pupils.

School factors and not solely pupil composition probably cause variation in smoking prevalence amongst schools, but there are no theoretical models to explain why. In this paper we propose a hypothesis to explain schools' influence on pupils' smoking and test this using an existing cross-sectional survey of 23,282 pupils from 166 secondary schools in the West Midlands, UK. We hypothesise that school-level educational achievement scores would not be associated with smoking prevalence, but schools providing value-added education given the social background of pupils (authoritative schools) would provide effective support and control, have a relatively strong influence on pupils' lives and be associated with lower than average smoking prevalence. Schools providing value-denuded education (laissez-faire schools) would have a relatively weak influence on pupils' lives and be associated with higher than average smoking prevalence. The school achievement measures were the proportion of pupils achieving 5A-C General Certificates of Secondary Education (5A-Cs) grades and the proportion of half days lost to truancy. Value-added/denuded terms were created by regressing 5A-Cs and truancy on five markers of the social profile of pupils at the school. Authoritative schools achieved better than expected rates on both measures. Laissez-faire schools achieved worse than expected rates on both measures. All other schools were classed as indeterminate. Multilevel logistic regression was used to relate the risk of regular smoking to school culture in both achievement and authoritative/laissez-faire terms, both with and without adjustment for pupil-level risk factors for smoking. As predicted, schools' achievement measures were unrelated to pupils' smoking. The odds ratios (95% confidence intervals) for smoking in authoritative and laissez-faire schools relative to indeterminate schools were 0.80 (0.70-0.91) and 1.16 (1.07-1.27), respectively. Adjustment for pupil-level smoking risk factors had little effect. School culture is an independent risk factor for adolescent smoking. Schools providing effective support and control might protect pupils from smoking.