Shyness is a temperamental trait characterized by an anxious preoccupation with the self and heightened reactivity in social situations. This trait is moderately stable across the childhood to adulthood [1]. Shy children often express social withdrawal and internalizing problems (i.e., loneliness, anxiety and depressive symptoms), if they have experienced negative peer experiences, such as peer rejection and victimization [2-6]. In contrast, mutual and high-quality friendships [6, 7] as well as parenting styles characterized by high warmth and autonomy [4, 8-10] buffer against social withdrawal and internalizing problems among shy youth. While the link between shyness and later socioemotional problems is well-documented, little is known about physical health outcomes of shy children even though epidemiologic studies suggest that childhood social withdrawal/isolation, an outcome of childhood shyness, is an independent risk factor for increased cardiovascular disease (CVD) risk in adulthood [11-13]. Similarly, studies using animal models show that rodents and dogs with stable neophobic/inhibited traits have shorter lifespan, dampened immune response, and elevated glucocorticoid production [14-17]. Yet, no studies in humans have identified whether early temperamental shyness plays a role in increased CVD risk in adulthood and how temperamental shyness, peer and parental relationships shape the development of physiological systems linked to CVD risk across the lifespan to confer later health risks. This research proposal aims to address these knowledge gaps by considering a broad range of negative and positive peer-experiences in relation to CV risk factors measured from childhood to adulthood, and the potential role of child mental health within this relationship. Using the ASLPAC cohort, we will address the following questions:

References
1. Pérez-Edgar, K. and N.A. Fox, Temperament and anxiety disorders. Child and Adolescent Psychiatric Clinics, 2005. 14(4): p. 681-706.
2. Boivin, M., S. Hymel, and W.M. Bukowski, The roles of social withdrawal, peer rejection, and victimization by peers in predicting loneliness and depressed mood in childhood. Development and Psychopathology, 1995. 7(4): p. 765-785.
3. Gazelle, H. and G.W. Ladd, Anxious solitude and peer exclusion: A diathesis–stress model of internalizing trajectories in childhood. Child development, 2003. 74(1): p. 257-278.
4. Booth-LaForce, C. and M.L. Oxford, Trajectories of social withdrawal from grades 1 to 6: Prediction from early parenting, attachment, and temperament. Developmental psychology, 2008. 44(5): p. 1298.
5. Tang, A., et al., Shyness trajectories across the first four decades predict mental health outcomes. Journal of abnormal child psychology, 2017. 45(8): p. 1621-1633.
6. Oh, W., et al., Trajectories of social withdrawal from middle childhood to early adolescence. Journal of Abnormal Child Psychology, 2008. 36(4): p. 553-566.
7. Fordham, K. and J. Stevenson-Hinde, Shyness, friendship quality, and adjustment during middle childhood. The Journal of Child Psychology and Psychiatry and Allied Disciplines, 1999. 40(5): p. 757-768.
8. Booth-LaForce, C., et al., Parent and peer links to trajectories of anxious withdrawal from grades 5 to 8. Journal of Clinical Child & Adolescent Psychology, 2012. 41(2): p. 138-149.
9. Kiel, E.J. and K.A. Buss, Prospective relations among fearful temperament, protective parenting, and social withdrawal: The role of maternal accuracy in a moderated mediation framework. Journal of abnormal child psychology, 2011. 39(7): p. 953-966.
10. Lewis‐Morrarty, E., et al., Infant attachment security and early childhood behavioral inhibition interact to predict adolescent social anxiety symptoms. Child development, 2015. 86(2): p. 598-613.
11. Caspi, A., et al., Socially isolated children 20 years later: risk of cardiovascular disease. Archives of Pediatrics & Adolescent Medicine, 2006. 160(8): p. 805-811.
12. Danese, A., et al., Adverse childhood experiences and adult risk factors for age-related disease: depression, inflammation, and clustering of metabolic risk markers. Archives of pediatrics & adolescent medicine, 2009. 163(12): p. 1135-1143.
13. Lacey, R.E., M. Kumari, and M. Bartley, Social isolation in childhood and adult inflammation: evidence from the National Child Development Study. Psychoneuroendocrinology, 2014. 50: p. 85-94.
14. Cavigelli, S.A., Behavioral Inhibition in Rodents: A Model to Study Causes and Health Consequences of Temperament, in Behavioral Inhibition. 2018, Springer. p. 35-58.
15. Cavigelli, S.A. and M. McClintock, Fear of novelty in infant rats predicts adult corticosterone dynamics and an early death. Proceedings of the National Academy of Sciences, 2003. 100(26): p. 16131-16136.
16. Corsetti, S., et al., Bold personality makes domestic dogs entering a shelter less vulnerable to diseases. PloS one, 2018. 13(3): p. e0193794.
17. Cavigelli, S.A., J.R. Yee, and M.K. McClintock, Infant temperament predicts life span in female rats that develop spontaneous tumors. Hormones and Behavior, 2006. 50(3): p. 454-462.

Pre-eclampsia is a common pregnancy complication and is one of the leading causes of death and injury to mothers and their babies worldwide. Pre-eclampsia occurs in the second half of pregnancy and is characterised by high blood pressure and protein in the urine. We do not fully understand why some women get this condition, but the function and leakiness of small blood vessels (capillaries) is important.

The glycocalyx, is a very thin gel-like layer which is found on the wall of all blood vessels. It seems to act as a barrier, controlling how much water remains inside the blood vessel. Researchers have recently started looking at the glycocalyx and have found in other conditions which cause leaky blood vessels (e.g. diabetes and kidney disease), the glycocalyx appears damaged.

Recently we have completed work to show that glycocalyx is not only present in blood vessels, but also the human placenta. We have done this by staining placenta with different proteins called lectins. These bind to the sugary glycocalyx, and when labelled with a fluorescent dye glow green under the microscope. We compare this green stain to a red membrane stain and can measure the depth of the glycocalyx.

We suspect that the glycocalyx is damaged in pre-eclampsia, like in other diseases.

ALSPAC offers an excellent resource of placental tissue, however, many of these specimens have been stored for a number of years and we know the glycocalyx can be a fragile structure. We propose a feasibility experiment, whereby the research team could look a small selection of ALSPAC placentas to examine if the glycocalyx has been preserved and can still be measured.

Poor quality diet is associated with increased risk of heart disease, stroke, type 2 diabetes, many forms of cancer and mental illness. Poor quality diet is the top contributor to mortality globally, and is estimated to cost the NHS £6billion per year.
The period of adolescence to early adulthood is the time when prevalence of overweight and obesity develops most rapidly, making this an important time to understand the contribution of diet to these developing risk factors. Adolescence and early adulthood is a time of rapid personal development and changing lifestyles. It also is the time when adult behaviours, including adult dietary patterns, are developed and established. Understanding the factors that influence development of adult diet is an important first step in developing strategies to change behaviour.
However, there has been little assessment of diet in young adults. Online tools (INTAKE24) are now available that make reporting on nutritional intake easier, allowing online completion of recalls of daily intake, and automated data processing. These validated tools make the collection of diet data in large samples more feasible, and are now being implemented in large-scale dietary surveys across many countries by researchers at the MRC Epidemiology Unit.
This application focusses on assessing the diet of young adults (age 28-30) in the ALSPAC cohort. This data will then be used together with other data collected within ALSPAC, or from linked datasets, to understand (1) the changes in environment and lifestyle which help to explain why individuals have adopted their current diets, and (2) the relationships between diet and measures of heart and blood health. The data will also be available for further research studying associations between diet at age 30 years and longer term measures of health and disease.

Mercury (Hg) is an environmental pollutant that can persist and bio-accumulates as methylmercury (MeHg) through the food chain. Foetuses are especially vulnerable to prenatal exposure since mercury can cross the placental barrier and the blood brain barrier is not fully developed until several months after birth. Prenatal exposure to Hg has been associated with impaired foetus development, such as reduced placental functioning and foetal growth. Prenatal exposure to Hg has also been associated with effects on child neuropsychological development. The specific mechanisms of toxicity related to these associations remain unclear, although some research has suggested that dramatic DNA methylation changes and epigenetic remodelling during early embryogenesis could be involved. Thus, cells and tissues acquire new methylation patterns that may persist in foetal development and childhood. To date, only three studies have been conducted relating Hg and epigenome-wide DNA methylation in cord blood with sample sizes between 138 and 321. These studies have identified altered expression in unique genomic regions as well as methylation changes in specific CpG sites. This study proposes to to investigate the association between prenatal Hg exposure and epigenome-wide methylation.

Maltreatment (physical, sexual and emotional abuse and neglect) in childhood is common and has both immediate and long-lasting negative effects. People who suffer maltreatment have a higher risk of many health conditions, including obesity, heart attack and stroke. However, it is not well understood why this is the case and at what age ill health starts to manifest. Unhealthy eating, smoking, physical inactivity and inadequate sleep may play a role in this relationship, but mental health issues, such as anxiety and depression, inflammation and other biological factors can also be involved. These factors might have separate effects but also act together, and these mechanisms might differ between men and women. This research aims to understand when the cardiovascular consequences of maltreatment appear, the pathways that link childhood maltreatment to heart disease in later life and whether these mechanisms differ by sex. This will help to identify potential targets for interventions to prevent heart disease in those who suffered maltreatment. We will use data from several British studies (ALSPAC, UK Biobank, the 1958 British Birth Cohort, Millennium Cohort and Growing up in Scotland), that have assessed maltreatment in early life and have measures of cardiovascular health indicators at different ages to understand when, why and how people who suffered maltreatment in childhood have a higher risk of heart disease in later life.

This proposal will look to obtain DNA samples from ALSPAC participant (G1) family members, including fathers and siblings.

A consensus of research has demonstrated that stress and adversity can become embodied and transmitted across generations, creating pathways by which social and economic inequality can affect human biology and health for decades. Most research has identified fetal development as a sensitive period for this transmission of stress from mother to child, with considerably less research on the postnatal period. Maternal postnatal stress has been found to shape infant stress response development, potentially creating a pathway by which maternal stress can become embodied in the next generation and influence how the next generation responds to and handles stressors. However, it is not clear how these effects on the stress response become embodied and whether these changes persist through childhood. This study proposes to test whether maternal postnatal stress in her child’s infancy and toddlerhood is related to the child’s methylation of stress-response related genes at age 7.

Similarly, previous research has demonstrated a relationship between psychosocial stress and childhood growth in weight and height. However, this relationship has been inconsistently demonstrated and the pathways by which stress affects growth are not clear. While the hypothalamic-pituitary-adrenal (HPA) axis has been implicated in this relationship, results have been inconsistent. This research project proposes to test the relationship between growth velocity throughout childhood and methylation of HPA-axis related genes at age 7, in order to determine whether alterations to stress response physiology are a mechanism by which stress can affect growth.

Although there are many common risk factors between non-alcoholic fatty liver disease (NAFLD) and cardiovascular disease, e.g. obesity, NAFLD may be associated with cardiovascular disease, independently from other established cardiovascular risk factors.