Cardiac arrest in paediatric patients often results in death or survival with severe brain injury. Therapeutic hypothermia, lowering of core body temperature to 32 to 34⁰C may reduce injury to the brain in the period after circulation has been restored. This thesis comprises studies related to the feasibility of performing a randomised controlled trial (RCT) of therapeutic hypothermia for neuroprotection after cardiac arrest in the UK. A systematic Cochrane review of paediatric evidence finds no published RCTs supporting or refuting the use of therapeutic hypothermia after cardiac arrest. Four on-going RCTs are identified which will add to the future evidence base; however, a future UK RCT is recommended. Additional support for a RCT is demonstrated by two UK surveys of paediatric intensive care and emergency care clinicians. Current UK practice is varied and clinical equipoise exists regarding post cardiac arrest temperature management. A national, retrospective study of all admissions to paediatric intensive care after out of hospital (OHCA) and in hospital cardiac arrest (IHCA) shows an overall survival of 76 and 50% respectively. Important differences between IHCA and OHCA populations are identified, recommending separation in a RCT. The incidence rate of cardiac arrest admissions to PICU in the UK is too low to recruit to a UK only RCT, after consideration of sample size requirements. A large, multi-centre, retrospective, observational study of OHCA patients identified multiple factors associated with survival. A survival prediction model, incorporating: pupillary reaction, blood lactate level and duration of cardiac arrest, is described. The model could be used as a tool for stratified randomisation within a RCT. Finally, therapeutic hypothermia is retrospectively compared with standard, normothermic temperature management after OHCA. In a limited population, no difference in survival is found; however, important information on application, logistics and safety of the intervention are evaluated.

Cerebral palsy (CP) affects around every one in 500 children born. It isn’t a particular illness or disease, but an umbrella term used to describe a physical condition that affects movement as a result of injury to the brain. There are several types of CP, the main ones being spastic, athetoid and ataxic. Despite medical advances, there is no cure for CP but there are ranges of treatments from drugs to Botulinum toxin type A injections, massage therapy to surgery. The aim of this study is to look at two of these treatments, namely Botulinum toxin type A injections and physiotherapy to treat spastic CP. Botulinum toxin is widely used to reduce muscle tone in the treatment of spasticity in children with cerebral palsy. The aim of the study is to compare the effects treatment with Botulinum toxin type A and Botulinum toxin type A with additional physical therapy in the management of a group of children with cerebral palsy. Experiments were done at The Prince Sultan Hospital and Al-Hada Armed Forces Hospital in Saudi Arabia. The local Ethics Committee approved the protocol. 47 children were recruited. All had cerebral palsy, diplegia, spasticity of the ankle planter flexors and significant gait abnormalities due to dynamic equinus foot deformity. They were divided into two groups. Both groups had their Gross Motor Function assessed one week before injection and at 4 and 6 weeks after injection. Additional measurements of range of movement and stiffness at the ankle and soleus electromyograms were recorded The soleus EMG was silent during ankle dorsiflexion in 20 children four weeks after injection of Botox. The EMG had returned six weeks after injection in every child. The Gross Motor Function Measurements were not significantly different in the two groups before the injection (p=0.23). The measurements improved significantly over the next six weeks in both groups (p<0.001). The magnitude of the improvement was greater in the group, which received Botulinum toxin type A and physical therapy (means 57.2 + 8.90 before, 64.9 + 9.78 after. Mean + SD) than in the group which received Botulinum toxin type A alone (59.5 + 11.0 before, 62.4 + 11.3 after Mean + SD). Conclusions 1. . The Treatment allocation provided groups, which were comparable pre-treatment in terms of baseline GMFM. 2. . Both treatments showed evidence of improvement in GMFM over the period of the study and particularly at 52 weeks. 3. . Treatment 2 showed a significant average advantage in GMFM over Treatment 1 at all times in the study. 4. . This advantage in average GMFM increased from 4 through to 52 weeks with a clear and significant difference between 4 and 52 weeks. 5. . This average advantage appeared to increase the higher the child’s baseline GMFM.

Since the introduction of glucocorticoids (GCs) in the treatment of rheumatoid arthritis in 1949, GC therapy has been associated with a number of adverse effects. Long-term use of GCs can result in growth retardation during childhood due to their actions on growth plate chondrocytes, although the exact mechanisms involved are unclear. The work of this thesis has investigated the cellular and molecular mechanisms involved in mediating GC effects at the growth plate. Affymetrix microarray has been used to identify and characterise the expression of lipocalin 2, a novel GC-responsive chondrocyte gene which may contribute to GC-induced growth retardation in the growth plate. In vitro and in vivo studies have also been used to examine the role of the cell cycle regulator, p21WAF1/CIP1 in GC-induced growth retardation. Finally, the growth plate sparing effects of a novel GC receptor modulator, AL-438, have also been identified. AL438, has reduced effects on bone growth compared to Dex, but maintains similar anti-inflammatory efficacy. This work has not only determined novel mechanisms of GC-induced growth retardation, but has also advanced the search for novel GC receptor modulators with reduced adverse effects.

Objective: The present study aimed to investigate the factors relating to mood and cognition which influence a person’s ability to participate in rehabilitation after Acquired Brain Injury (ABI). It was hypothesised that impairment in cognition, including specific impairment in executive functioning and depression would be associated with poorer engagement in rehabilitation. Method: Twenty-nine patients undergoing rehabilitation following stroke (89.7%) or TBI (10.3%) participated. Individuals recruited completed the Hospital Anxiety and Depression Scale as a measure of mood and an executive functioning test battery. Data collection occurred over a two week period as concurrent ratings of participation were gathered from physiotherapists and occupational therapists using the Pittsburgh Rehabilitation Participation Scale. Results: In support of the hypotheses, correlation analysis showed a significant positive correlation between participation in rehabilitation with executive functioning (p < .05) and a significant negative correlation between participation in rehabilitation and low mood (p < .05). No association was found between general cognitive ability, functional disability, time since injury, age, gender and participation.

Objectives: To study autism spectrum disorder (ASD) in the Faroe Islands, including prevalence, diagnostic stability and environmental factors that are potentially involved in the aetiology of autism. Method: I. The target group was recruited from the entire population sample of participants with ASD during a two-phase screening and diagnostic process of the entire Faroe Islands population in the relevant school age group born between 1985-1994 (7-16 years, n=7,689) in 2002 and again in 2009 (15-24 years, n= 7,128) using an independent clinical diagnosis and standardised tools. II. The diagnostic stability of ASD from childhood to early adulthood over a period of 7 years compared diagnoses in 2002 and 2009. III. A literature search of vitamin D and ASD covering the period from January 1 1995 to October 31 2011 was carried out. IV. A pilot study involved questioning 20 mothers of young individuals from the target group and 13 mothers of healthy comparisons, regarding mothers’ diet habits, health, life-style and well-being during their pregnancy with an index child. V. 25-hydroxyvitamin D3 (25(OH)D3) levels were examined in a population based cross-sectional study that involved 219 individuals: 40 participants with a diagnosis of ASD from the target group (31 males/9 females), their 62 typically developing siblings (29 brothers/33 sisters), their 77 parents (40 mothers/37 fathers), and 40 healthy comparisons (28 males/12 females). Results: I. The rate of ASD rose significantly from 0.56% (n=43) in 2002 to 0.93% (n=66) in 2009. Although these results were still within the range of typical findings from other studies, of the 24 newly discovered cases in 2009 nearly half were females thus altering the male/female ratio from 6/1 to 2.7/1. II. The stability of clinical ASD diagnosis was perfect for AD, good for “atypical autism”/PDD-NOS, and less than perfect for Asperger syndrome (AS). Stability of the diagnoses made by means of research tools were more variable but still good for AD. Both systems showed excellent stability over the seven-year period for “any ASD” diagnosis, although a number of clear cases (especially in females) had been missed in the original screening in 2002. These results support the notion that a single overarching diagnostic category, ‘autism’ or ASD, would better suit clinical realities as outlined in the new DSM-5. III. The systematic review (in 2010) provided some, albeit very limited, support for the possible role of vitamin D deficiency in the pathogenesis of ASD: there are three main areas of involvement of vitamin D in the human body that could potentially have direct impact on the development of ASD: (1) the brain, (2) gene regulation and (3) the immune system. The prevalence of ASD has been suggested to be raised at higher latitudes. IV. Mothers of individuals with ASD had had during their pregnancy significantly less positive “attitude to sun” (p=0.001), consumed fewer vegetables (p=0.026) and also less fruit (p=0.078). V. The ASD case group had significantly lower 25(OH)D3 levels (24.8 nmol/L) than their typically-developing siblings (42.6 nmol/L, p<0.001) and their parents (44.9 nmol/L, p<0.001), and also significantly lower than healthy age and gender matched comparisons (37.6 nmol/L, p=0.002). There was a trend for males having lower 25(OH)D3 levels than females. There was no association between vitamin D and age, month/season of birth, IQ or subcategories of ASD. Among the ASD group, 60% were severely deficient (<30 nmol/L) and 84.2% of the whole study sample (n=219) had deficient/insufficient levels (<50/<75 nmol/L). Conclusions: I. ASD prevalence in the Faroe Islands increased from 0.56% in 2002 to 0.93% in 2009 mainly due to missed cases in 2002, nearly half of them females. II. There was diagnostic stability for the overall category of ASD over time in the group diagnosed in childhood (7—16) years, but considerable variability with regards to diagnostic sub-groupings. Diagnosing females require novel approach. III. Vitamin D deficiency–either during pregnancy or early childhood–may be an environmental trigger for ASD in individuals genetically predisposed to the broad phenotype of autism. IV. There are some interesting differences in the diet and life-style habits between mothers with a child with ASD and mothers with a healthy child. The ASD-group’s negative “attitude to sun” may indicate some life-style/health differences which may play a role in pathogenesis of ASD, especially in combination with other environmental risk factors. V. The present study, demonstrating an association between low levels of 25(OH)D3 and ASD, is the first to be based in a total population and to use siblings, parents and general population control groups. It adds to similar findings from other regions of the world, indicating vitamin D deficiency in the population and especially in individuals with ASD. As all groups were exposed to low levels of sunlight, the very low 25(OH)D3 in the ASD group suggests that some other underlying pathogenic mechanism may be involved.

In the absence of readily available physiological models of human growth, the effects of oestradiol on the human C28/I2 chondrocyte cell line were studied. The classical oestrogen receptors, ERα and ERβ, were shown to be expressed in both murine and human chondrocyte cell lines. Oestradiol and related chemicals, which alter the function of the oestrogen receptors (ER), were exploited to tease out the different functions of each ER in the growth plate. In the absence of foetal bovine serum, oestradiol had no effect on proliferation, differentiation or apoptosis of chondrocyte cells in monolayer culture or on the growth of the foetal metatarsal culture system. In addition, oestradiol did not convey a protective effect on chondrocytes exposed to the pro-inflammatory cytokines, tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in monolayer culture. However, endogenous oestrogen appears to play an important role in maintaining chondrocyte proliferation in monolayer culture and mineralisation in metatarsal culture as reflected by the inhibitory effects of Faslodex, the non-specific ER antagonist, on chondrocytes and metatarsals in culture. In the presence of methyl-piperidino-pyrazole (MPP), a selective ERα antagonist, and raloxifene, a selective oestrogen receptor modulator with higher ERβ binding affinity, a reduction in chondrocyte proliferation and increase in apoptosis was observed in murine and human chondrocytes. Similarly, a marked reduction in linear growth occurred when foetal murine metatarsals were exposed to MPP and raloxifene in combination. A less marked reduction in growth was observed in MPP-treated metatarsals. These findings suggest that the oestrogen receptors may have opposing actions in the growth plate with ERβ acting like a brake on chondrocyte growth and ERα promoting growth. ERβ may regulate cell proliferation through control of cell cycle modulators affecting G1/S phase transition as MPP and raloxifene in combination reduced cyclin E and p53 levels on Western blot analysis. The aim of the second part of my thesis was to investigate the effect of oral oestrogen on linear growth in girls with primary ovarian insufficiency (POI). A retrospective review of girls with POI treated at a tertiary endocrinology clinic over an 11 year period was performed. As expected the majority of girls with POI had Turner syndrome (TS; 83.7%). Non-TS associated POI was rare and the leading cause was iatrogenic secondary to the effects of total body irradiation for bone marrow transplantation (12.8%). A significant proportion of these girls developed POI after full pubertal development so few cases were available to investigate the effect of oestrogen on growth. The oral oestrogen regime followed in individual patients with TS was highly variable so it was not possible to assess the effects of dose on height velocity or bone maturation in this retrospective audit. However, the second clinical study examined in detail the effect of oestrogen on growth in TS girls who received a standardised course of oral ethinylestradiol for pubertal induction and a standard dose of growth hormone (10 mg/m2/week). These girls participated in a prospective randomised double-blind placebo-controlled multi-centre study of growth promoting treatment in TS. The girls were initially randomised to oxandrolone or placebo at 9 years of age and further randomised to oral ethinylestradiol at 12 or 14 years of age. The results of this study are embargoed until published. The laboratory effects of oestradiol found in this thesis suggest that ERα may stimulate or maintain growth, and ERβ may inhibit growth. The obvious question is how these observations might be involved in the complex relationship between puberty, oestrogen and height velocity in humans. As affinity studies show that the half maximal effective concentration (EC50) of ERα is achieved at slightly lower concentrations of oestradiol than ERβ it is conceivable that the ERα effect could predominate at lower systemic oestradiol concentrations and that ERβ could become more important at higher concentrations for example in later puberty. Alternatively, it is possible that the expression of ERα reduces or ERβ increases in the growth plate after reaching peak height velocity.

Background: Reducing neonatal mortality rates [NMR] (deaths/1,000 live births within 28 days of delivery) is a key global health goal. Using comparable data from Ghana (West Africa) and Scotland, I investigated NMR, specific causes of death and risk factors in the two countries. By identifying the main causes of excess mortality in Ghana and where they occur, it is hoped more effective strategies can be developed. Methods: This thesis used a multilevel mixed methods study design. Data on live births were obtained from three Health and Demographic Surveillance Systems (HDSS) in the north, middle and south of Ghana respectively: Navrongo (2004-12; 17,016 live births, 320 deaths); Kintampo (2005-10; 11,207 live births, 140 deaths); Dodowa (2006-14; 21,647 live births, 135 deaths). Comparable Scottish data were obtained from the Information Services Division (1992 to 2015; 1,278,846 live births, 2,783 deaths). Each dataset was analysed by neonatal death (dead/alive), using univariate and multivariable logistic regression. The multivariable analyses adjusted for maternal demographic and obstetric characteristics. Missing data were analysed using multiple imputation techniques. Data analyses were complemented by a researcher-developed questionnaire survey of 71 maternity care providers in the three regions of Ghana followed by face-to-face in-depth interviews with 48 maternity care providers who had experience of prematurity, birth asphyxia, neonatal infection and neonatal death. Results: The NMRs in the three HDSS were: Navrongo: 18.8; Kintampo: 12.5; and Dodowa 6.2 and in Scotland it was 2.2; the NMR in both countries is reducing. More than 99% of the neonatal deaths in Scotland occurred in the first week compared to 74% in Ghana. The leading causes of neonatal deaths (NMR) in Ghana were infection (4.3), asphyxia (3.7) and prematurity (2.2). In Scotland, they were congenital malformations (0.6), asphyxia (0.4) and prematurity (0.3). Only 88 deaths (0.07) of neonatal deaths in Scotland were due to infection. Ninety-eight percent of babies born in Scotland were born in a health facility compared to 60% of babies born in Ghana (hospital: 38.1%; clinic: 21.1%). In Ghana, babies born in hospitals had a higher risk of neonatal mortality compared to those born at home (NMR-hospital: 15.6; clinic: 7.1; home: 11.8). Most of the neonatal deaths in Ghana occurred at home (54%); there were more deaths among babies who were born in a hospital but died at home (hosp/home) compared to those born at home but died in a hospital (home/hosp). Asphyxia was the leading cause of death among hosp/hosp, and infection was the leading cause of death among hosp/home, home/home and home/hosp. Neonatal mortality in Ghana was largely influenced by where mothers sought maternity service, or the type of personnel who provided maternity care service. Mothers and babies who were cared for in hospitals by doctors and midwives received relatively better care and proper management of birth complications. Those who were cared for in clinics received basic delivery services and management of uncomplicated asphyxia. Mothers and babies who were cared for at home by traditional birth attendants (TBA) received poor care and poor management of neonatal illnesses based on traditional approaches which increased the risk of death. Women’s maternity choices were influenced by wider societal factors including prominent cultural values, family hierarchical structures and the cost of maternity services, and individual/ family factors including place of residence and availability of transport and beliefs about the cause of disease. Conclusion: There is considerable opportunity for reducing NMR in Ghana, especially deaths due to asphyxia and infections. Most uncomplicated deliveries should be performed by midwives in community clinics. The number of community maternity clinics should gradually be increased to enable home deliveries by TBAs to be phased out. Facilities should be improved for delivery and postnatal care in hospitals and the proportion of sick babies managed by health care workers trained in their care should be increased. Regular postnatal checks in the community by trained staff should be standard.

Childhood onset growth hormone deficiency (CO-GHD) may contribute to low bone mass and alterations to body composition. This thesis consists of a series of studies utilising dual-energy X-ray absorptiometry (DXA), peripheral quantitative computerized tomography (pQCT) and biochemical assessment of bone health and body composition of CO-GHD. In addition, metabolic profiles, glucose metabolism as well as quality of life have been studied in these subjects. Furthermore, an interventional study of weight bearing exercise (WBE) was performed to explore its role in influencing the bone health of children and adolescents with CO-GHD. Chapter 1, relevant literature reviews explore: bone structure, growth, development and strength; GH/IGF-1 system and its actions; CO-GHD and its impacts during childhood and transition; and WBE and its mechanism and impacts on bone health. Chapter 2 presents the rationale and specific aims of this thesis. Chapter 3, a retrospective multicentre review of management of young adults with CO-GHD in four paediatric centres in Scotland during transition. Medical records of 130 eligible CO-GHD adolescents (78 males), who attained final height between 2005-2013 were reviewed. Of the 130, 74/130(57%) had GH axis re-evaluation by stimulation tests /IGF-1 measurements. Of those, 61/74(82%) remained GHD with 51/74(69%) restarting adult rhGH. Predictors of persistent GHD included an organic hypothalamic-pituitary disorder and multiple pituitary hormone deficiencies (MPHD). Despite clinical guidelines, there was significant variation in the management of CO-GHD in young adulthood across Scotland. Chapter 4, a cross-sectional control study of bone DXA measurements in (n=21) subjects with CO-GHD treated with rhGH and had attained final height from 2005 to 2013 in a single tertiary paediatric centre compared to (n= 21) heights/age matched healthy controls. By applying different models of DXA adjustment, our analysis revealed lower TB-BMC for bone area in males with CO-GHD and lower LS-BMAD SDS in females with CO-GHD compared to matched controls. In addition, subjects with CO-GHD had lower LM for height and higher FM for height compared to controls, and this was more pronounced in males than females (p=0.04). The time of onset and aetiology of CO-GHD have a larger influence on accrual of bone mass in these patients. These findings indicate that adolescents with CO-GHD have a low bone mass, despite prior long term rhGH replacement therapy. In chapter 5, we investigated bone health of subjects with CO-GHD at time of initial evaluation and retesting at final height. A total of 25 children (first time assessment group) undergoing GH stimulation tests for investigation of short stature (naive GHD-15, normal-10), and 11adolescents with CO-GHD (retesting group) undergoing biochemical re-evaluation at final height after withdrawal of rhGH therapy (persistent GHD-7, GH-sufficient-4) were recruited from Royal Hospital for Children between 2012-2013. By using further bone health assessment methods in addition to DXA (including p.QCT, mechanography, bone profiles and biomarkers), the bone density and body composition did not differ when we compared GHD to matched height but normal GH at initial evaluation and retesting. However, naive GHD had lower muscle force as assessed by mechanography compared to the normal. In addition, bone resorption biomarker CTX was significantly higher in naive GHD vs. normal and that was significantly correlated to PTH levels in both first time assessment and retesting groups. Our results suggest that muscle force and serum PTH may be important determinants of bone health in subjects with CO-GHD. Chapter 6 investigates lipids, adipokines (leptin- adiponectin- resistin) and glucose homeostasis and their relationship with bone and body composition in children and adolescents with CO-GHD at times of initial evaluation and retesting at final height (same population as chapter 5). Lipid profiles, adipokines and glucose homeostasis were not different between those with GHD and those who had normal GH levels across the groups of first time assessment and retesting. In the retesting group, those who were older at the time of diagnosis of CO-GHD with a shorter duration of rhGH therapy were more likely to have higher cholesterol(r=0.9, p&lt;0.001), leptin (r=0.8, p&lt;0.001), and lower osteoclacin (r=-0.7, p=0.01) at final height. Leptin levels correlated positively with osteocalcin at diagnosis (r=0.51, p=0.01) but inversely at retesting (r=-0.91, p&lt;0.01). The conclusion was that the timing and duration of childhood rhGH therapy might influence adiposity parameters and bone metabolism in subjects with CO-GHD. In chapter 7 the study participants of chapter 5 were asked to complete either Short Form-36 (SF-36) or Adult Growth Hormone Deficiency Assessment (AGHDA) quality of life (QoL) questionnaires at the time of assessment of their GH axis. Our analysis showed that the overall QoL was not altered in children with naive GHD with a total score of SF-36 [93 (77, 96) naive GHD vs. 90 (84, 93) normal, P=0.56] (higher scores reflect better QoL). However, naive GHD had less energy and vitality scores compared with normal (75 (65, 100) vs. 95 (65,100) respectively, p=0.04), when the normal scored lower in the subscale of emotional well-being compared to those with naive GHD (78 (55, 84) vs. 90 (68, 96) respectively, p&lt;0.001). In the retesting group, those with persistent GHD scored better in the AGHDA than GH sufficient (6 points (2, 8) vs. 9 points (7, 17) respectively, though not significant (p= 0.10) (higher scores reflect poorer QoL). Unexpectedly, subscale analysis showed that GH-sufficient subjects significantly lacked energy and complained of tiredness compared to those who were confirmed to have persistent GHD (5 points (3, 6) vs. 1 point (0, 1) respectively, p= 0.03). Further studies to validate QoL specific instruments in this population are needed with greater insight to elucidate factors that modify the relationship between GH status and QoL in children and adolescents. Chapter 8 was a prospective intervention, randomised controlled study of 14 subjects among the first time assessment group (GHD-10, normal-4) and five subjects with CO-GHD among retesting group (persistent GHD-4, GH-sufficent-1). Subjects were randomised into either an exercise intervention group (EX) (25 jumps off 25 cm platform step/three days/week for six months) or a control, in addition to rhGH being prescribed. The results of this study were limited by the small sample size and poor compliance. Therefore, there were insufficient data to recommend the use of weight bearing exercise in the absence of rhGH in children and adolescents with CO-GHD. Further studies with adequate sample size that can more rigorously exam the optimal exercise interventions are needed. Chapter 9 discusses the main findings of each chapter in this thesis and outlines potential limitations of the thesis methodology, and some important and interesting areas for future research in children and adolescents with CO-GHD.