Assessing the immune phenotype of cardiac syndrome x: a prospective study of biomarkers

Cardiac Syndrome X (CSX), the presence of angina pectoris with objective signs of myocardial ischaemia despite angiographically normal epicardial coronary arteries, appears to be due to coronary microvascular dysfunction and is known to be associated with an elevation of several inflammatory biomarkers, suggesting a possible role for inflammation in its pathogenesis. We aimed to further characterise this relationship by prospectively analysing a wide variety of molecular biomarkers in a cohort of CSX patients thereby charting the changes in biomarkers throughout the natural history of CSX from its initial diagnosis to eventual disease quiescence. We found that CSX patients, when compared to healthy controls, have a persistent low-grade systemic inflammatory response characterised by an elevation of Tumour Necrosis Factor and Interferon-gamma, regardless of the presence of contemporaneous signs or symptoms of disease activity. Interleukin-6 and C-reactive Protein (CRP) are only elevated when patients have clinical evidence of disease activity and may be state markers in CSX. Moreover, CRP levels appear to correlate with signals of disease severity such as the time taken to develop symptoms during exercise stress testing. We have also demonstrated that the enzyme Indoleamine-2,3- dioxygenase is upregulated in active disease thus providing a possible explanation for the increased burden of psychological disease encountered in CSX. Analysis of the microRNA transcriptome showed that miR-143 is significantly under-expressed in CSX patients. This could allow phenotype switching in vascular smooth muscle cells with the resultant vascular remodelling causing reduced vessel responsiveness to local rheological stimuli and reduced luminal diameter with consequent increased microvascular resistance during times of increased myocardial oxygen demand, thereby limiting maximal hyperaemia during exercise. Our findings corroborate many previous hypotheses regarding the role of inflammation in CSX, generate new insights into possible pathogenic mechanisms and offer new therapeutic targets for the future management of this important cardiological condition.

The gut microbiota as a contributing factor to antipsychotic-induced weight gain and metabolic dysfunction

Schizophrenia represents one of the world’s most devastating illnesses due to its often lifelong course and debilitating nature. The treatment of schizophrenia has vastly improved over recent decades with the discovery of several antipsychotic compounds; however these drugs are not without adverse effects that must be addressed to maximize their therapeutic value. Newer, atypical, antipsychotics are associated with a compilation of serious metabolic side effects including weight gain, insulin resistance, fat deposition, glucose dysregulation and ensuing co-morbidities such as type II diabetes mellitus. The mechanisms underlying these side effects remain to be fully elucidated and adequate interventions are lacking. Further understanding of the factors that contribute these side effects is therefore required in order to develop effective adjunctive therapies and to potentially design antipsychotic drugs in the future with reduced impact on the metabolic health of patients. We investigated if the gut microbiota represented a novel mechanism contributing to the metabolic dysfunction associated with atypical antipsychotics. The gut microbiota comprises the bacteria that exist symbiotically within the gastrointestinal tract, and has been shown in recent years to be involved in several aspects of energy balance and metabolism. We have demonstrated that administration of certain antipsychotics in the rat results in an altered microbiota profile and, moreover, that the microbiota is required for the full scale of metabolic dysfunction to occur. We have further shown that specific antibiotics can attenuate certain aspects of olanzapine and risperidone–induced metabolic dysfunction, in particular fat deposition and adipose tissue inflammation. Mechanisms underlying this novel link appear to involve energy utilization via expression of lipogenic genes as well as reduced inflammatory tone. Taken together, these data indicate that the gut microbiota is an important factor involved in the myriad of metabolic complications associated with antipsychotic therapy. Furthermore, these data support the future investigation of microbial-based therapeutics for not only antipsychotic-induced weight gain but also for tackling the global obesity epidemic.

Preclinical atherosclerosis in rheumatoid arthritis

Introduction: There is accumulating evidence of an increased risk of cardiovascular morbidity and mortality in rheumatoid arthritis patients. A combination of both traditional cardiovascular risks and rheumatoid specific factors appear to be responsible for driving this phenomenon. Rheumatoid arthritis has been an orphan of cardiologists in the past and rheumatologists themselves are not good at CVD screening. Identifying the extent of preclinical atherosclerosis in RA patients will help us to appreciate the magnitude of this serious problem in an Irish population. Methods: We undertook a cross-sectional study of 63 RA patients and 48 OA controls and compared the 2 groups with respect to 1) traditional CV risks factors, 2) serum biomarkers of inflammation, including CRP, TNFα, IL6 and PAI-1, 3) carotid intima-media thickness (cIMT), carotid plaque and ankle-brachial index (ABI) as markers of pre-clinical atherosclerosis, 4) biochemical and ultrasonic measures of endothelial dysfunction and 5) serum and echocardiographic measures of diastolic dysfunction. Within the RA group, we also investigated for associations between markers of inflammation, subclinical atherosclerosis and diastolic dysfunction. Results: Prevalence of traditional CV risks was similar in the RA and OA groups. A number of biomarkers of inflammation were significantly higher in the RA group: CRP, fibrinogen, IL- 2, -4, -6, TNFα. PAI-1, a marker of thrombosis, correlated with disease activity and subclinical atherosclerosis in RA patients. With regard to subclinical atherosclerosis measures, RA patients had a significantly lower ABI than OA patients. Carotid plaque and cIMT readings were similar in RA and OA patients. Assessment of endothelial function revealed that RA patients had significantly higher concentrations of adhesion molecules, in particular sero-positive RA patients and RA smokers. Adhesion molecule concentrations were associated with markers of diastolic dysfunction in RA. Urine PCR, another marker of endothelial dysfunction also correlated with diastolic dysfunction in RA. Assessment of endothelial function with flow mediated dilatation (FMD) found no difference between the RA and OA groups. Disease activity scores in RA patients were associated with endothelial dysfunction, as assessed by FMD. Conclusions: We did not find significant differences in measures of subclinical atherosclerosis, flow mediated dilatation or diastolic function between RA and OA patients. This is most likely in part due to the fact that there is increasing evidence that OA has an inflammatory component to its pathogenesis and is associated with metabolic syndrome and increased CV risk. We reported a significant association between urinary PCR and measures of diastolic dysfunction. Urinary PCR may be a useful screening tool for diastolic dysfunction in RA. The association between RA disease activity and measures of vascular function supports the theory that the excess cardiovascular burden in RA is linked to uncontrolled inflammation.

Exploring the relationship between patterns of physical activity, sedentary behaviour and cardiometabolic health in middle-aged Irish adults

Background and Study Rationale Being physically active is a major contributor to both physical and mental health. More specifically, being physically active lowers risk of coronary heart disease, high blood pressure, stroke, metabolic syndrome (MetS), diabetes, certain cancers and depression, and increases cognitive function and wellbeing. The physiological mechanisms that occur in response to physical activity and the impact of total physical activity and sedentary behaviour on cardiometabolic health have been extensively studied. In contrast, limited data evaluating the specific effects of daily and weekly patterns of physical behaviour on cardiometabolic health exist. Additionally, no other study has examined interrelated patterns and minute-by-minute accumulation of physical behaviour throughout the day across week days in middle-aged adults. Study Aims The overarching aims of this thesis are firstly to describe patterns of behaviour throughout the day and week, and secondly to explore associations between these patterns and cardiometabolic health in a middle-aged population. The specific objectives are to: 1 Compare agreement between the International Physical Activity Questionnaire-Short Form (IPAQ-SF) and GENEActiv accelerometer-derived moderate-to-vigorous (MVPA) activity and secondly to compare their associations with a range of cardiometabolic and inflammatory markers in middle-aged adults. 2 Determine a suitable monitoring frame needed to reliably capture weekly, accelerometer-measured, activity in our population. 3 Identify groups of participants who have similar weekly patterns of physical behaviour, and determine if underlying patterns of cardiometabolic profiles exist among these groups. 4 Explore the variation of physical behaviour throughout the day to identify whether daily patterns of physical behaviour vary by cardiometabolic health. Methods All results in this thesis are based on data from a subsample of the Mitchelstown Cohort; 475 (46.1% males; mean aged 59.7±5.5 years) middle-aged Irish adults. Subjective physical activity levels were assessed using the IPAQ-SF. Participants wore the wrist GENEActiv accelerometer for 7 consecutive days. Data was collected at 100Hz and summarised into a signal magnitude vector using 60s epochs. Each time interval was categorised based on validated cut-offs. Data on cardiometabolic and inflammatory markers was collected according to standard protocol. Cardiometabolic outcomes (obesity, diabetes, hypertension and MetS) were defined according to internationally recognised definitions by World Health Organisation (WHO) and Irish Diabetes Federation (IDF). Results The results of the first chapter suggest that the IPAQ-SF lacks the sensitivity to assess patterning of activity and guideline adherence and assessing the relationship with cardiometabolic and inflammatory markers. Furthermore, GENEActiv accelerometer-derived MVPA appears to be better at detecting relationships with cardiometabolic and inflammatory markers. The second chapter examined variations in day-to-day physical behaviour levels between- and within-subjects. The main findings were that Sunday differed from all other days in the week for sedentary behaviour and light activity and that a large within-subject variation across days of the week for vigorous activity exists. Our data indicate that six days of monitoring, four weekdays plus Saturday and Sunday, are required to reliably estimate weekly habitual activity in all activity intensities. In the next chapter, latent profile analysis of weekly, interrelated patterns of physical behaviour identified four distinct physical behaviour patterns; Sedentary Group (15.9%), Sedentary; Lower Activity Group (28%), Sedentary; Higher Activity Group (44.2%) and a Physically Active Group (11.9%). Overall the Sedentary Group had poorer outcomes, characterised by unfavourable cardiometabolic and inflammatory profiles. The remaining classes were characterised by healthier cardiometabolic profiles with lower sedentary behaviour levels. The final chapter, which aimed to compare daily cumulative patterns of minute-by-minute physical behaviour intensities across those with and without MetS, revealed significant differences in weekday and weekend day MVPA. In particular, those with MetS start accumulating MVPA later in the day and for a shorted day period. Conclusion In conclusion, the results of this thesis add to the evidence base regards an optimal monitoring period for physical behaviour measurement to accurately capture weekly physical behaviour patterns. In addition, the results highlight whether weekly and daily distribution of activity is associated with cardiometabolic health and inflammatory profiles. The key findings of this thesis demonstrate the importance of daily and weekly physical behaviour patterning of activity intensity in the context of cardiometabolic health risk. In addition, these findings highlight the importance of using physical behaviour patterns of free-living adults observed in a population-based study to inform and aid health promotion activity programmes and primary care prevention and treatment strategies and development of future tailored physical activity based interventions.

Unconjugated bile acids influence expression of circadian genes: a potential mechanism for microbe-host crosstalk

Disruptions to circadian rhythm in mice and humans have been associated with an increased risk of obesity and metabolic syndrome. The gut microbiota is known to be essential for the maintenance of circadian rhythm in the host suggesting a role for microbe-host interactions in the regulation of the peripheral circadian clock. Previous work suggested a role for gut bacterial bile salt hydrolase (BSH) activity in the regulation of host circadian gene expression. Here we demonstrate that unconjugated bile acids, known to be generated through the BSH activity of the gut microbiota, are potentially chronobiological regulators of host circadian gene expression. We utilised a synchronised Caco-2 epithelial colorectal cell model and demonstrated that unconjugated bile acids, but not the equivalent tauro-conjugated bile salts, enhance the expression levels of genes involved in circadian rhythm. In addition oral administration of mice with unconjugated bile acids significantly altered expression levels of circadian clock genes in the ileum and colon as well as the liver with significant changes to expression of hepatic regulators of circadian rhythm (including Dbp) and associated genes (Per2, Per3 and Cry2). The data demonstrate a potential mechanism for microbe-host crosstalk that significantly impacts upon host circadian gene expression. Disruptions to circadian rhythm in mice and humans have been associated with an increased risk of obesity and metabolic syndrome. The gut microbiota is known to be essential for the maintenance of circadian rhythm in the host suggesting a role for microbe-host interactions in the regulation of the peripheral circadian clock. Previous work suggested a role for gut bacterial bile salt hydrolase (BSH) activity in the regulation of host circadian gene expression. Here we demonstrate that unconjugated bile acids, known to be generated through the BSH activity of the gut microbiota, are potentially chronobiological regulators of host circadian gene expression. We utilised a synchronised Caco-2 epithelial colorectal cell model and demonstrated that unconjugated bile acids, but not the equivalent tauro-conjugated bile salts, enhance the expression levels of genes involved in circadian rhythm. In addition oral administration of mice with unconjugated bile acids significantly altered expression levels of circadian clock genes in the ileum and colon as well as the liver with significant changes to expression of hepatic regulators of circadian rhythm (including Dbp) and associated genes (Per2, Per3 and Cry2). The data demonstrate a potential mechanism for microbe-host crosstalk that significantly impacts upon host circadian gene expression.