Subclinical Disease Research Articles

Incorporating a polygenic risk score into cardiovascular disease examinations to identify subclinical coronary artery disease: rationale and design of the ESCALATE trial

Abstract Background Identifying and targeting established modifiable risk factors such as hyperlipidaemia, blood pressure, smoking, and diabetes have led to a successful reduction in poor outcomes from coronary artery disease (CAD) at the population-level. However, up to one-in-four patients who present with ST elevation myocardial infarction do so in the absence of such risk factors. There is a significant unmet need for new diagnostic strategies independent of these factors to detect CAD in its subclinical phase. Polygenic risk scores (PRS) have demonstrated an ability in cohort studies to improve risk prediction models independent of traditional risk factors and self-reported family history. However, there is a lack of data from implementation and interventional studies demonstrating efficacy, participant and provider experience, and cost effectiveness of utilising PRS. Aim The aim of this study is to examine the incorporation of a CAD PRS during primary care-based cardiovascular disease (CVD) risk assessments to identify low or intermediate risk individuals with newly diagnosed CAD and subsequent impact on shared treatment decisions, participant experience, and provider experience. Our hypothesis is that incorporating a CAD PRS-based decision tool for a coronary calcium scan into CVD risk assessments will identify new, potentially missed individuals with subclinical CAD. Methods The ESCALATE study is a 12-month, prospective, multi-centre implementation study incorporating CAD PRS into otherwise standard primary care CVD risk assessments, identifying patients at increased lifetime CAD risk, for non-invasive CT coronary imaging. 1,000 eligible participants aged 45-65 years old will enter the PPP-CAD Diagnostic Pathway, which applies CAD PRS to those considered low or moderate 5-year absolute CVD risk based on traditional risk factors and triages those with CAD PRS ≥80% for a coronary calcium scan (Figure 1). The primary outcome will be the identification of subclinical CAD, defined as a coronary artery calcium score (CACS) >0 Agatston units (AU). Multiple secondary outcomes will be assessed, including baseline CACS ≥100 AU or ≥75th age-/sex-matched percentile, medical management strategies, cholesterol and blood pressure levels, health-related quality of life (HRQOL), and patient experience. Results Study recruitment was initiated recruitment in August 2022 and is actively recruiting across Australia. 298 participants have been enrolled as of 27th February 2022. Conclusion The ESCALATE study is the first prospectively registered clinical trial testing the ability of a CAD PRS to: a) identify individuals with subclinical CAD by non-invasive coronary imaging; and b) measure the impact of this diagnostic strategy on personalised diagnosis and shared decision-making. The top-line study results are expected in mid-2025.

Plasma levels of amyloid beta 1-40 are associated with ultrasonographic morphological characteristics related with carotid plaque vulnerability in individuals without cardiovascular disease

Abstract Background Amyloid-beta (1-40) (Αb40), a proinflammatory and pro-atherosclerotic peptide, is associated with concurrent subclinical atherosclerotic cardiovascular disease (ASCVD) and with major adverse cardiac events in both primary and secondary prevention settings conferring additive and reclassification value over traditional risk factors and risk scores. This evidence supports the hypothesis that Ab40 could be a biomarker of ASCVD. However, mechanistic data on the association of Αb40 with in vivo morphological characteristics of atherosclerosis related with plaque vulnerability are not available. Purpose To examine the association of Ab40 levels with characteristics of the carotid artery wall in the general population. Methods In the settings of the Athens Vascular registry, subjects without clinically overt ASCVD were consecutively recruited (n=301). Subclinical carotid atherosclerosis was assessed by ultrasonography and the images were subsequently analyzed using a dedicated software (AMS v3.03). The sum and maximal area of atherosclerotic plaques (PLQ) and the minimum echogenicity using grey scale median (GSM) of intima-media thickness (IMT, IMT_GSM) and atherosclerotic plaques (PLQ_GSM) were used as the main endpoints of the analysis. Αb40 was measured in plasma samples. Results Subjects with Ab40 levels in the highest tertile had higher prevalence of male sex, presence of carotid plaque atherosclerosis, higher sum and maximal PLQ are and lower GFR HDL-C levels and lower PLQ_GSM (p<0.05 for all). Ab40 in the highest vs. lower tertile was associated with higher sum and maximal plaque area (b-coefficient= 22.9 95% CI: 0.72, 45.1, p=0.43 and b-coefficient= 10.5 95% CI: 1.77, 19.3, p=0.019, for sum and maximal plaque area, respectively) after adjustment for traditional cardiovascular risk factors and GFR (core model). Moreover, Ab40 in the highest tertile was associated with decreased GSM of both IMT and PLQ in the carotid arteries (b-coefficient= -6.0 95% CI: -9.6, -2.36, p=0.001, for IMT and b-coefficient= -8.66 95% CI: -16.5, -0.79, p=0.31, for PLQ) after adjustment for the core model. Conclusion In individuals without clinically overt ASCVD, higher levels of Ab40 are associated with the larger atherosclerotic burden and with ultrasonographic characteristics related to higher plaque vulnerability. These findings suggest a mechanistic background for the established association of Ab40 with major adverse cardiovascular events.

Menopausal vasomotor symptoms and subclinical atherosclerotic cardiovascular disease

Abstract Background Menopausal vasomotor symptoms (VMS) have been associated with subclinical and manifest atherosclerotic cardiovascular disease (ASCVD) but the scientific basis is limited and VMS have not been studied in relation to image-detected coronary atherosclerosis. PURPOSE To assess, in peri- and postmenopausal women, associations between VMS and subclinical ASCVD detected through computed tomography angiography (CCTA), coronary artery calcium score (CACS) or carotid ultrasound, and if potentially determined associations are modified by menopausal hormone therapy (MHT) or regular physical activity (PA). Methods This cross-sectional population-based study was conducted on a subset of participants from the Swedish CArdioPulmonary BioImage Study (SCAPIS), including women 50-65 years of age. The women underwent comprehensive cardiovascular assessments and completed an extensive questionnaire, which included questions about current and previous menopause-related symptoms. VMS was assessed on a 4-point scale and analyzed in relation to subclinical ASCVD, detected via CCTA, CACS and carotid ultrasound using multivariate logistic regression analyses. Results Of 2995 women included, 14.2% reported severe VMS (n= 425), 18.1% moderate VMS (n=543), and 67.7% no or mild VMS (n=2027). Women with severe, but not those with moderate, VMS had higher prevalence of CCTA detected atherosclerosis (34.1% vs 27.8%, p<0.05), but not CACS >100 (5.8% vs 6.6% p >0.05) or any carotid plaque (47.6% vs 46.6%, p>0.05) than women with no or mild VMS. Using the same reference category, severe but not moderate VMS was significantly associated with CCTA-detected atherosclerosis, odds ratio (OR) before and after multivariable adjustment 1.36, 95% confidence interval (CI) 1.08 – 1.72 and 1.33, 95% CI 1.02 – 1.72, respectively. This association became stronger when requiring an over 5-year duration of severe VMS, multivariable adjusted OR 1.53, 95% CI 1.09 – 2.14. Additional adjustment for MHT strengthened the association (OR 1.64, 95% CI 1.16 – 2.32). No significant association with CACS >100 or with the presence of any carotid plaque was observed. Additional adjustment for PA did not change any associations. Conclusions Severe but not moderate VMS were significantly associated with CCTA detected atherosclerosis, independent of a broad range of cardiovascular risk factors. Further research is required to determine the eventual causality of this relationship.Figure 1.Forest plots

Association of proteomic signatures and immunophenotyping profiles with subclinical cardiovascular disease characterised by multimodal imaging in South African systemic lupus erythematosus patients

Abstract Background Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease associated with autoantibodies (aAb). Cardiovascular disease (CVD) is a leading cause of mortality later in the disease course. Protein signatures may have utility in predicting SLE patients at risk of CVD. Purpose To determine the association of aAb protein signatures and plasma markers of immune activation with subclinical CVD in South African SLE patients characterised by multimodal imaging. Methods SLE patients, without known CVD, were recruited from rheumatology clinics. Early cardiovascular (CV) involvement was characterised using carotid ultrasound, echocardiography and cardiovascular magnetic resonance (CMR). Commercially available ELISA and Luminex multibead platforms profiled plasma immunophenotyping and CV biomarkers including IL1B, IL6, TNFa, IFNg, MMP9, TIMP1, ICAM1, VCAM1, adiponectin, galectin3, ST2, NTproBNP, hsTropT and hsCRP. We investigated IgG and IgA aAb responses on 2 protein microarrays, first analysing pooled samples on the i-Ome array of over 1600 proteins and second, individual patient samples were analysed on an autoimmune disease array of 50 autoantigens. Raw data were imported and analysed using R. First, loess normalisation of proteomic data was conducted to remove any systematic experimental variation, followed by exploratory analyses on normalised data, then permutational multivariate analysis of variance (PERMANOVA) assessed which imaging variables were most influential on aAb profiles and these results were then further analysed using linear modelling. Results 18 SLE patients and 6 controls were included in this exploratory analysis. Mean age was 34±9 years, 89% were women and the most frequent CV risk factors were hypertension, smoking, and dyslipidaemia in 28%, 28% and 17%, respectively. On CMR, 72% of SLE patients were found to have subclinical CVD, predominantly myocardial (72%) and pericardial (22%) involvement. Compared to controls, SLE patients had significantly increased TNFa (p=0.046), VCAM1 (p=0.006), adiponectin (p=0.046) and NTproBNP (p=0.04). On IgG assay proteomic analyses, PERMANOVA results showed that impaired peak longitudinal strain (p=0.03) and the presence of any regional myocardial oedema (T2 ratio >1,9) (p=0.025) were the most influential variables related to peptide aAb profiles, and late gadolinium enhancement (LGE) was the most influential variable related to protein aAb profiles (p=0.075). Linear modelling considering peak longitudinal strain, T2 ratio (Figure 1) and LGE identified a list of 5, 9 and 13 candidate biomarkers respectively for these CMR parameters. Conclusions In South African SLE patients with a high burden of subclinical CVD associated with elevated immunophenotyping markers, we identified imaging parameters that were influential on protein signatures. Linear modelling identified a list of candidate biomarkers associated with CMR indicators of myocardial strain, oedema and fibrosis.Candidate biomarkers

Discovering pericoronary adipose tissue differences between normal and atherosclerotic coronary segments with radiomics analysis

Abstract Introduction Pericoronary adipose tissue (PCAT) extracted from coronary CT angiography (CCTA) is a novel marker for coronary inflammation. Establishing correlation between PCAT changes and the vascular lesion itself is essential for using PCAT to monitor clinical and subclinical disease status. Past studies focused on patient-level and vessel-level PCAT differences. However, lesion-level PCAT differences beyond mean attenuation values are not well understood. Purpose To investigate the differences between PCAT adjacent to atherosclerotic plaques and PCAT adjacent to normal coronary segments using radiomics analysis. Material and Methods This study enrolled 100 consecutive patients who underwent CCTA. A total of 446 plaques were successfully annotated in 89 patients. The PCAT of these 446 plaques and also their normal segment counterparts were extracted. Following prior literature, adipose tissue was defined as voxels with attenuation ranging -190 to -30 HU. PCAT was extracted within 3 mm distance from the vessel wall, along the middle 10 mm of each plaque. The PCAT of normal coronary artery segments were extracted both proximal and distal with distance D mm to the plaque with 10 mm length. We used 3 different distances to define the normal segments (D={10, 20, 30}). PyRadiomics was used to generate radiomics features and the random forest classifier from Scikit-Learn was used to distinguish PCAT extracted adjacent to atherosclerotic plaques and PCAT adjacent to normal coronary segments. To minimize bias due to vessel diameter or shape and focus on the texture of PCAT, we included only the First Order, GLCM, GLDM, GLRLM, GLSZM and NGTDM features (total 93 features) from PyRadiomics. We used 5-fold cross validation splitted at patient-level to develop the classifier. Result The radiomics model achieved acceptable discrimination for D=30 mm (mean AUROC = 0.70), borderline discrimination for D=20 mm (mean AUROC=0.68) and D=10 mm (mean AUROC = 0.60). These results suggest a trend toward greater differences between PCAT further away from the lesion compared with PCAT adjacent to the lesion. The feature "Gray Level Size Zone Matrix (GLSZM): Low Gray Level Zone Emphasis" had the highest importance in 4 out of 5 folds in our D = 30 mm experiment. The mean value of this feature is significantly different for the PCAT adjacent to the lesions compared to PCAT extracted from the normal segments (0.152 vs 0.166, p=0.001). Conclusion We investigated the difference between PCAT adjacent to atherosclerotic plaques and PCAT adjacent to normal coronary segments using radiomics analysis. We found greater discrimination when the normal segments were extracted further away from the lesion, suggesting PCAT changes due to atherosclerosis may exist beyond the length of the plaque. Moreover, "Gray Level Size Zone Matrix (GLSZM): Low Gray Level Zone Emphasis" could serve as a novel radiomic biomarker to correlate PCAT changes to atherosclerosis.

Abstract 13302: Associations of Allostatic Load With Brain Macro-Morphology, Subclinical Cerebrovascular Disease, and Microstructural Integrity of White Matter Pathways: The Boston Puerto Rican Health Study

Introduction: Allostatic load (AL), a measure of overall body wear and tear, has been associated with structural and functional alterations of the brain, but the evidence remains scarce. We aimed to evaluate the prospective associations of AL with brain MRI measures in the Boston Puerto Rican Health Study (BPRHS). Research Questions: Whether AL is associated with 1) brain macro-morphology: brain age and volumes, 2) subclinical cerebrovascular disease: white matter hyperintensity (WMH) and brain infarcts, and 3) microstructural integrity of white matter (WM) pathways in Puerto Rican adults. Methods: Adults from the BPRHS cohort who provided blood and urine samples at baseline, 2-y, and 6-y visits and underwent MRI scans at 13-y visit were included in analyses (n=226). AL was evaluated as a composite score (0-11) of 11 physiological parameters in five body systems. An overall AL score was calculated by averaging the scores of the baseline, 2-y, and 6-y visits, with higher scores indicating greater dysregulation. We evaluated brain age and volumes using T1-weighted MRI, WMH and brain infarcts using T2-weighted MRI, and microstructural integrity of WM pathways using diffusion tensor imaging. Multivariable logistic regression models were used to examine the associations between AL and brain infarcts, and linear regression models were used for other MRI outcomes. Results: Mean age at baseline was 54.6 ± 6.6 y, with 79.2% female. Higher AL was associated with higher brain age z scores (β = 0.129, P = 0.031) and lower volumes of total WM, cerebral WM (especially parietal and temporal lobes), cerebellum, and brain stem. Higher AL was also associated with higher grades of WMH (β = 0.207, P = 0.024) and higher odds of small vessel infarcts [OR (95% CI) = 3.50 (1.56-7.87), P = 0.002], but not large vessel infarcts [OR (95% CI) = 1.33 (0.73-2.40), P = 0.348]. Diffusion tensor imaging parameters showed that participants with higher AL had more microstructural deterioration patterns in major WM pathways. Conclusions: Higher AL score was associated with brain macro- and microstructural disruptions and a higher risk of subclinical cerebrovascular disease, suggesting that AL may be a potential indicator of the need for early intervention in preventing neuronal damage in older adults.

Abstract 15088: Presence and Severity of Subclinical Atherosclerotic Disease in the Primary Prevention of the Oncologic Population

Introduction: There is scarce evidence about endothelial dysfunction and damage in patients with a history of cancer in cardiovascular primary prevention and the impact in the development of Subclinical Atherosclerotic Disease (SCAD). Methods: A cross-sectional study was carried out on a registry of patients enrolled in primary prevention who had a Doppler ultrasound (DUS) screening for SCAD in the carotid and ileofemoral territory from 09/2020 to 04/2023. Inclusion criteria: 18 years old or older. Exclusion criteria: Previous cardiovascular and/or cerebrovascular event. History of cancer was defined as active or remitted cancer, excluding basal cell and squamous cell carcinoma. Population was divided in two, G1: non-oncologic group; G2 oncologic group. SCAD was defined as the presence of more than 1 atherosclerotic plaque in the carotid and/or ileofemoral territory. Univariate and bivariate analysis were done to establish population characteristics. Results: A total of 7920 patients were included: 7694 in G1 and 226 in G2 (97.15% vs 2.85%). G2 patients were older, mostly women and had higher prevalence of hypertension and dyslipidemia. Clinical characteristics and findings of the DUS are shown in Table1. A linear regression was done to assess the total plaque area compared to oncologic history, adjusted for age, sex, smoking history, dyslipidemia, obesity, sedentarism, diabetes, and hypertension. Results show that history of cancer is a predictor for SCAD (OR=1.43 [IC 95%: 1.1-1.9], p=0.021). Cancer also seems to be associated with a higher number of territories affected by SCAD (OR=1.63 [IC 95%: 1.2-2.1], 0.001) adjusted for age, sex, tobacco use, obesity, dyslipidemia, diabetes, sedentarism, and hypertension. Conclusions: Patients with history of cancer had more SCAD. Cancer behaved as a predisposing factor for increased plaque presence and area, adjusted for classic risk factors. This raises the need for more intensive control of SCAD in cancer patients.

Abstract 15643: Social Determinants of Health, Epigenetic Aging, and Subclinical Cardiovascular Disease: The Coronary Artery Risk Development in Young Adults (CARDIA) Study

Introduction: Associations of social determinants of health (SDOH) with subclinical cardiovascular disease (CVD), such as left ventricular mass index (LVMI) and coronary artery calcification (CAC), are poorly described. Mechanisms of such associations may involve epigenetic changes like DNA methylation. We sought to examine associations between selected SDOH measures and subclinical CVD, and potential mediation by GrimAge acceleration (GrimAA), a measure of epigenetic aging. Methods: We included 2932 Black and White adults from the CARDIA year 20 (Y20) exam (mean age 45y, 48% Black, 57% female). Associations between SDOH measures at Y20 (or cumulative up to Y20) and Y25 ln(CAC score +1) or LVMI (in g/m 2.7 ) were examined using linear regression. Statistical mediation analysis was used to estimate the proportion of associations attributable to GrimAA among 880 participants in whom it was measured. Results: All selected SDOH measures at Y20 - residential racial segregation (G* Score); census tract poverty; food insecurity; difficulty paying for basics, medications, or healthcare; and maximum years of participant or parental education - were associated with LVMI at Y25 (Table). For example, each 1% higher census tract poverty was associated with an average 0.95 g/m 2.7 higher LVMI. Several SDOH measures were also associated with CAC score at Y25. Similar associations were observed between cumulative SDOH exposures from Y0-Y20 and Y25 subclinical CVD. GrimAA mediated >30% of the statistical association between participant education and LVMI or CAC (Table), and >20% of the associations of residential segregation and census tract poverty with LVMI. Conclusions: SDOH metrics were significantly and longitudinally associated with subclinical CVD in midlife. DNA methylation, as represented by GrimAge acceleration, appears to statistically mediate these associations, suggesting possible molecular mechanisms by which SDOH may accelerate CVD risk.

Abstract 12041: Coronary Artery Calcium and All-Cause Mortality in the Multicenter Aids Cohort Study: MACS

Introduction: People living with HIV (PLWH) have more subclinical cardiovascular disease than people without HIV (PWOH), but few studies have evaluated risk for mortality based on coronary artery calcium (CAC) among this population. Purpose: This study aimed to determine the association between CAC and all-cause mortality among male PLWH and PWOH and to identify a potential interaction with HIV serostatus. Methods: The study population was derived from the MACS, a US prospective observational cohort study including male PLWH and PWOH. Participants underwent cardiac non-contrast computed tomography from July 2004 to November 2013. Men with atrial fibrillation, coronary heart disease, or coronary revascularization were excluded. Cox proportional hazards model was used to calculate adjusted hazard ratios [aHR] for all-cause mortality among men with vs. without baseline CAC (Agatston score>0) and per standard deviation (SD) increment in continuous Agatston score [log (CAC+1)], controlling for demographic and cardiac risk factors. CAC differences by HIV serostatus were evaluated using multiplicative CACхHIV interaction terms. Results: Among 1344 males (mean age 50 years, CAC prevalence 46%, 823 (61%) PLWH), we observed 110 deaths (13%) among PLWH and 41 deaths (8%) among PWOH during the follow-up period (median: 13.4 years). Among PLWH, 62% had an undetectable plasma HIV viral load, the median CD4 cell nadir was 284 cells/μL, and 17% had a history of clinical AIDS. The age-adjusted mortality rate was 13.5 (95% CI: 11.0-16.5) among PLWH and 7.8 (5.4-10.9) among PWOH per 1000 person-years. CAC presence was associated with all-cause mortality among all participants (aHR=1.47, 95% CI: 1.02-2.11, p=0.04), and among PLWH (aHR=1.61, 1.04-2.47, p=0.03). In PWOH, we found no significant association (aHR=1.27, 0.63-2.59, p=0.50), although the interaction was not significant (p= 0.49). Higher log (CAC+1) was also associated with all-cause mortality among all participants (aHR=1.37 per SD, 1.15-1.63, p<0.001) and among strata (PLWH, aHR=1.40, 1.13-1.73, p=0.002; and PWOH aHR=1.38, 1.00-1.91, p=0.049). Conclusion: CAC predicts all-cause mortality in a large cohort of PLWH and PWOH, with no significant difference in this association by HIV serostatus.

Abstract 12439: Abdominal Visceral Fat Area and Density Are Associated With Coronary Artery Calcification: A Cross-Sectional Analysis of the Multi-Ethnic Study of Atherosclerosis

Introduction: The associations of body composition components, including muscle and adipose tissue, and markers of subclinical coronary artery disease are unclear. We examined the relationship between abdominal CT-derived measures of the area and density of fat and muscle with coronary artery calcification (CAC), using data from the Multi-Ethnic Study of Atherosclerosis (MESA). Methods: A randomly selected total of 1,974 MESA participants free of coronary heart disease underwent abdominal CT scans at exams 2 or 3, with the resulting images interrogated for abdominal body composition. Using six cross-sectional slices spanning L2-L5, Medical Imaging Processing Analysis and Visualization (MIPAV) software was used to determine abdominal muscle and fat composition by utilizing appropriate Hounsfield units (HU) ranges. CT chest scans obtained during the exam 1 were used to obtain CAC scores, calculated using the Agatston method and spatially weighted calcium score (SWCS). Multivariable linear regression analysis was performed to assess the relationship between both abdominal visceral fat and muscle area and density to prevalent CAC. Results: A total of 1,089 participants had a CAC>0, with an average CAC score of 310. In the fully adjusted model, for every 10 cm 2 increase in visceral fat area, the likelihood of having a CAC >0 increased by 0.60% (p<0.001). In the minimally adjusted model, abdominal muscle area was significantly associated with CAC>0, which became non-significant in the fully adjusted model. For density of visceral fat , every 1 HU increase (less lipid dense fat tissue), the likelihood of having a CAC score>0 decreased by 0.29% (p<0.05). No significant relationship was observed between density of abdominal muscle and CAC>0. Conclusion: Greater area and higher lipid density of abdominal visceral fat was associated with an increased likelihood of having CAC, while there was no significant relationship between abdominal muscle area or density and CAC. Both the quantity and the quality of fat have associations with an important marker of subclinical atherosclerosis, CAC, and their significance with respect to cardiovascular outcomes requires further evaluation.

Abstract 15400: Association of Cardiovascular Health With Early Carotid Artery Changes in Young Adults From Disadvantaged Social Backgrounds: The Future of Families and Child Wellbeing Study (FFCWS)

Introduction: Associations of cardiovascular health (CVH), as measured by the AHA’s Life’s Essential 8 (LE8) score, with early markers of arterial injury have not been described in young adults. Methods: The FFCWS enrolled 4898 mother-baby dyads at delivery in 20 US cities in 1998-2000, oversampling mothers who were unmarried, including a disproportionate number of minority and low-income families. In 2021-23, offspring were invited to attend an in-person exam to measure CVH and perform carotid artery ultrasound. Individual CVH metric and overall LE8 scores (except sleep) were calculated for each participant (ppt). Carotid imaging was performed by 2 trained sonographers and images were analyzed by a core lab. We examined cross-sectional associations of LE8 scores with measures of carotid intima-media thickness (IMT), gray scale (a measure of early arterial injury), and distensibility, using linear regression. Results: Among 1211 offspring ppts, mean age was 22y, >50% self-identified as female, and >75% as Black or Hispanic. Black and Hispanic ppts had lower, and ppts from Other racial/ethnic groups had higher, overall LE8 scores compared with White ppts (all P≤0.01; Table 1A). In unadjusted analyses, Black ppts had thicker carotid IMT and lower Young’s elastic modulus measures. Prevalence of diagnostic plaque was low. In analyses adjusted for age, sex, and race/ethnicity (Table 1B), LE8 scores were significantly associated with lower carotid IMT, higher gray scale median, and lower Young’s elastic modulus. For example, for each 10 point higher (better) LE8 score, mean-maximal carotid IMT was lower by 0.008 mm. Conclusions: Among young adults largely from disadvantaged social backgrounds, the LE8 score was associated with early measures of carotid arterial injury. LE8 may be an important surrogate measure for monitoring long-term CVH and may help identify young adults with emerging subclinical disease and higher long-term cardiovascular risk.

Restless legs syndrome, periodic limb movements of sleep, and subclinical cardiovascular disease.

The online version contains supplementary material available at 10.1007/s41105-023-00497-7.

The problem of timely diagnosis of primary hyperaldosteronism. Clinical case

A clinical case is presented of delayed diagnosis of primary hyperaldosteronism (PHA) due to aldosterone-producing adenoma. Suspicion of PHA arises in case of persistent combination of hypertension with hypokalemia of various severity, a specific neuromuscular syndrome, as well as a high risk of cardiomyopathy, heart failure, and deterioration of kidney function, which are not typical for banal hypertension. Unsatisfactory blood pressure control with a standard combination of pharmacotherapy, an unexpected positive effect of mineralocorticoid receptor antagonists, high blood pressure in young people, as well as a family history of early high blood pressure or stroke at a young age, paroxysmal course of hypertension itself increases the risk of PHA. For a long time, a disease in our patient was considered a banal hypertension, although the diagnostic hypothesis of PHA should appear immediately after severe neuromuscular syndrome, rapid progression of cardiomyopathy and clinically significant heart failure began to dominate in the clinical picture. A 44-year-old female patient came to the attention of rheumatologists with suspicion of inflammatory myopathy due to frequent episodes of muscle weakness whose origin remained unclear. Key complaints of pronounced paroxysmal generalized muscle weakness, especially in the limbs, numbness and paresthesias in the extremities, calf muscle cramps against the background of hypertension were regarded as a neuromuscular syndrome specific to PHA. The PHA hypothesis was supported by the presence of cardiomyopathy, heart failure, clinically significant hypokalemia, and elevated aldosterone level. Computed tomography of the retroperitoneal space confirmed the presence of adrenal adenoma. Unfortunately, with a delay, after 13 years of hypertension, the diagnosis of PHA against the background of aldosterone-producing adenoma was confirmed. Consultation with a surgeon endocrinologist was suggested, as well as administration of eplerenone 50 mg daily in combination with lercanidipine 20 mg daily. Already after 6 weeks of pharmacotherapy, a positive therapeutic effect was obtained regarding the control of hypertension, heart failure, and the severity of neuromuscular syndrome. The differential diagnosis of secondary endocrine hypertension can be successful only with the possession of skills for early clinical detection of endocrine pathology, even in subclinical disease presentation. The diagnosis of PHA makes it possible to offer a patient radical surgical treatment, as well as to choose optimal approaches to pharmacotherapy.

Frequency of subclinical interstitial lung disease in COVID-19 autopsy cases: potential risk factors of severe pneumonia

Risk factors of severe coronavirus disease 2019 (COVID-19) have been previously reported; however, histological risk factors have not been defined thus far. The aim of this study was to clarify subclinical hidden interstitial lung disease (ILD) as a risk factor of severe pneumonia associated with COVID-19. We carefully examined autopsied lungs and chest computed tomography scanning (CT) images from patients with COVID-19 for interstitial lesions and then analyzed their relationship with disease severity. Among the autopsy series, subclinical ILD was found in 13/27 cases (48%) in the COVID-19 group, and in contrast, 8/65 (12%) in the control autopsy group (p = 0.0006; Fisher’s exact test). We reviewed CT images from the COVID-19 autopsy cases and verified that subclinical ILD was histologically detectable in the CT images. Then, we retrospectively examined CT images from another series of COVID-19 cases in the Yokohama, Japan area between February–August 2020 for interstitial lesions and analyzed the relationship to the severity of COVID-19 pneumonia. Interstitial lesion was more frequently found in the group with the moderate II/severe disease than in the moderate I/mild disease (severity was evaluated according to the COVID-19 severity classification system of the Ministry of Health, Labor, and Welfare [Japan]) (moderate II/severe, 11/15, 73.3% versus moderate I/mild, 108/245, 44.1%; Fisher exact test, p = 0.0333). In conclusion, it was suggested that subclinical ILD could be an important risk factor for severe COVID-19 pneumonia. A benefit of these findings could be the development of a risk assessment system using high resolution CT images for fatal COVID-19 pneumonia.

Modelling the overlap and divergence of autistic and schizotypal traits on hippocampal subfield volumes and regional cerebral blood flow

Psychiatric disorders show high co-morbidity, including co-morbid expressions of subclinical psychopathology across multiple disease spectra. Given the limitations of classical case-control designs in elucidating this overlap, new approaches are needed to identify biological underpinnings of spectra and their interaction. We assessed autistic-like traits (using the Autism Quotient, AQ) and schizotypy - as models of subclinical expressions of disease phenotypes and examined their association with volumes and regional cerebral blood flow (rCBF) of anterior, mid- and posterior hippocampus segments from structural MRI scans in 318 and arterial spin labelling (ASL) in 346 nonclinical subjects, which overlapped with the structural imaging sample (N = 298). We demonstrate significant interactive effects of positive schizotypy and AQ social skills as well as of positive schizotypy and AQ imagination on hippocampal subfield volume variation. Moreover, we show that AQ attention switching modulated hippocampal head rCBF, while positive schizotypy by AQ attention to detail interactions modulated hippocampal tail rCBF. In addition, we show significant correlation of hippocampal volume and rCBF in both region-of-interest and voxel-wise analyses, which were robust after removal of variance related to schizotypy and autistic traits. These findings provide empirical evidence for both the modulation of hippocampal subfield structure and function through subclinical traits, and in particular how only the interaction of phenotype facets leads to significant reductions or variations in these parameters. This makes a case for considering the synergistic impact of different (subclinical) disease spectra on transdiagnostic biological parameters in psychiatry.

Atherosclerosis of the iliac arteries for the prediction of erectile dysfunction and epistaxis in men undergoing abdominal CT scan

BackgroundTo investigate the association between erectile dysfunction (ED) as well as epistaxis (ES) in relation to the extent of iliac atherosclerosis.MethodsIn this retrospective cross-sectional study, all consecutive male patients treated at our institution from 01/2016 to 12/2020 undergoing abdominal CT scan were evaluated. Patients (n = 1272) were invited by mail to participate in the study in returning two questionnaires for the evaluation of ED (IIEF-5) and ES. Patients who returned filled-in questionnaires within a 3-month deadline were included in the study. The extent of atherosclerosis in the common iliac artery (CIA) and the internal iliac artery (IIA) was assessed by calcium scoring on unenhanced CT. Stratification of results was performed according to reported IIEF-5 scores and consequential ED groups.ResultsIn total, 437 patients (34.4% of contacted) met the inclusion criteria. Forty-two patients did not fulfill predefined age requirements (< 75 years) and 120 patients had to be excluded as calcium scoring on nonenhanced CT was not feasible. Finally, 275 patients were included in the analysis and stratified into groups of “no-mild” (n = 146) and “moderate-severe” (n = 129) ED. The calcium score (r=-0.28, p < 0.001) and the number of atherosclerotic lesions (r=-0.32, p < 0.001) in the CIA + IIA showed a significant negative correlation to the IIEF-5 score, respectively. Patients differed significantly in CIA + IIA calcium score (difference: 167.4, p < 0.001) and number of atherosclerotic lesions (difference: 5.00, p < 0.001) when belonging to the “no-mild” vs. “moderate-severe” ED group, respectively. A multivariable regression model, after adjusting for relevant baseline characteristics, showed that the number of atherosclerotic CIA + IIA lesions was an independent predictor of ED (OR = 1.05, p = 0.036), whereas CIA + IIA calcium score was not (OR = 1.00031, p = 0.20). No relevant correlation was found between ES episodes and IIEF-5 scores (r=-0.069, p = 0.25), CIA + IIA calcium score (r=-0.10, p = 0.87) or number of atherosclerotic CIA + IIA lesions (r=-0.032, p = 0.60), respectively.ConclusionsThe number of atherosclerotic lesions in the iliac arteries on nonenhanced abdominal CT scans is associated with the severity of ED. This may be used to identify subclinical cardiovascular disease and to quantify the risk for cardiovascular hazards in the future.Trial registrationBASEC-Nr. 2020 − 01637.

Evaluation of a method to measure the ratio of pelvic limb to thoracic limb girth in dogs

Objective To determine if a landmark-incorporated limb girth measurement in sound dogs, utilising the greater trochanter and the acromion process, would be both reliable and have a consistent ratio between pelvic limb (PL) and thoracic limb (TL) muscle measurements. Background To establish a reliable and reproducible reference range for sound dogs. This reference range may help clinicians further evaluate dogs during lameness and musculoskeletal examinations. Evidentiary value Prospective study of 115 sound dogs measured by a single observer. Methods Examinations were performed by one observer, using a Gullick II tape measure for PL to TL ratio (PL:TL) measurements for 115 dogs. Bodyweight, breed, body condition score (BCS), and sex were recorded. Each limb was measured three times. The average PL:TL per dog was calculated. Further statistical analysis was used to calculate intra-observer variance and the correlation of limb girth to body weight, BCS, and sex, with significance set at P &lt; 0.05. Results The average PL:TL of the sample was 1.515 ± 0.049. Fifty-two dogs of the 115 cases (45%) had a PL:TL ratio of 1.500. PL:TL measurements were not related to dog weight, BCS, or sex. The intra-class correlation was reported to be 0.99. Conclusion Our study suggests that a landmark-incorporated measurement in a weight-bearing position can be reproducible. Further investigation is required to determine if this measurement can be reproducible between multiple observers. Application A landmark-incorporated limb girth measurement may guide clinicians in case progression and help pinpoint subclinical musculoskeletal disease in dogs.

Genomic diversity and natural recombination of equid gammaherpesvirus 5 isolates

BackgroundEquid gammaherpesvirus 5 (EHV5) is closely related to equid gammaherpesvirus 2 (EHV2). Detection of EHV5 is frequent in horse populations worldwide, but it is often without a clear and significant clinical impact. Infection in horses can often present as subclinical disease; however, it has been associated with respiratory disease, including equine multinodular pulmonary fibrosis (EMPF). Genetic heterogeneity within small regions of the EHV5 glycoprotein B (gB) sequences have been reported and multiple genotypes of this virus have been identified within individual horses, but full genome sequence data for these viruses is limited. The primary focus of this study was to assess the genomic diversity and natural recombination among EHV5 isolates. ResultsThe genome size of EHV5 prototype strain and the five EHV5 isolates cultured for this study, including four isolates from the same horse, ranged from 181,929 to 183,428 base pairs (bp), with the sizes of terminal repeat regions varying from 0 to 10 bp. The nucleotide sequence identity between the six EHV5 genomes ranged from 95.5 to 99.1%, and the estimated average nucleotide diversity between isolates was 1%. Individual genes displayed varying levels of nucleotide diversity that ranged from 0 to 19%. The analysis of nonsynonymous substitution (Ka > 0.025) revealed high diversity in eight genes. Genome analysis using RDP4 and SplitsTree programs detected evidence of past recombination events between EHV5 isolates. ConclusionGenomic diversity and recombination hotspots were identified among EHV5 strains. Recombination can drive genetic diversity, particularly in viruses that have a low rate of nucleotide substitutions. Therefore, the results from this study suggest that recombination is an important contributing factor to EHV5 genomic diversity. The findings from this study provide additional insights into the genetic heterogeneity of the EHV5 genome.

Worms and welfare: Behavioural and physiological changes associated with gastrointestinal nematode parasitism in lambs

Parasitism with gastrointestinal nematodes (GIN) is a worldwide issue impacting negatively on animal production, health, and welfare. Therefore, early diagnostic signs of parasitism are required to allow for timely interventions. The objective of this study was to evaluate the behavioural and physiological changes in lambs associated with GIN infection. We used 30, 8-month-old Romney-cross wethers, that were administered anthelmintics until faecal egg counts (FEC) were zero and housed in an indoor facility. The study lasted 9 weeks, which comprised a 3-week pre-treatment, and a 6-week treatment phase. Lambs were randomly assigned to one of two treatments (n = 15/treatment) trickle-dosed with: 1) 1500 infective third stage larvae (L3) three days/week for 6 weeks (27,000 total L3; challenged), or 2) water 3 days/week for 6 weeks (control). Within each pen there were 5 pairs of lambs (balanced for liveweight), with each pair comprising a challenged and control lamb. Blood, faecal, and saliva samples were collected 1 week pre-treatment and weekly for 6 weeks of treatment. Behaviour was observed (e.g., feeding, lying, standing) from video-camera recordings using scan sampling every 5 min for 8 h, 1 day pre-treatment and on the day immediately prior to physiological sampling across the 6-week treatment phase (7 days in total). Accelerometers were attached to each lamb to continuously monitor behaviour from 3 weeks pre-treatment and for the remainder of the study. Liveweight, body condition, faecal soiling and faecal consistency scoring were performed weekly as was lipidomic analysis of plasma samples. From week 2 of treatment, challenged lambs spent less time feeding and more time lying than control lambs until week 5 of treatment (P ≤ 0.01). At week 3 of treatment, elevated lipids (mainly triglycerides and phospholipids), loose faeces and faecal soiling around the anus were observed in challenged lambs compared with controls (P ≤ 0.05). From week 4 of treatment, FEC were elevated in the challenged compared to control lambs (P ≤ 0.05). There was also lower liveweight gain at 4 and 5 weeks of treatment in the challenged lambs compared with control lambs (P ≤ 0.05). These results show a clear timeline of changes in behaviour (e.g., feeding and lying), lipids such as triglycerides, and digestive function (e.g., faecal soiling) suggestive of GIN subclinical disease, which show promise for use in future studies on early identification of subclinical GIN parasitism in lambs.

Clinical features and long-term survival in idiopathic pulmonary arterial hypertension with thyroid dysfunction: insights from a national multicentre prospective study.

Our objective was to clarify the characteristics and long-term survival of idiopathic pulmonary arterial hypertension (IPAH) patients with thyroid dysfunction and compare them with IPAH without thyroid dysfunction. A retrospective analysis was conducted using prospectively collected data. IPAH patients with thyroid dysfunction at baseline were included. Patients with other subgroups of PAH and Group 2-5 pulmonary hypertension were excluded. IPAH patients with euthyroid function were matched 1:1 to IPAH patients with thyroid dysfunction by age and sex. In total, 148 IPAH patients with thyroid dysfunction were included. Patients with hyperthyroidism, hypothyroidism, subclinical hyperthyroidism and subclinical hypothyroidism accounted for 16.2%, 18.9%, 8.1% and 56.8%, respectively. IPAH patients with hyperthyroidism showed the highest mixed venous oxygen saturation (SvO2) and the lowest pulmonary vascular resistance (PVR) at baseline among subgroups, while patients with subclinical hypothyroidism had the lowest SvO2 and highest PVR (p<0.05). Compared with IPAH without thyroid dysfunction, patients with hyperthyroidism (9.14 versus 13.86 WU; p<0.05) and hypothyroidism (10.70 versus 13.86 WU; p<0.05) showed significantly lower PVR. The haemodynamic profiles of patients with subclinical hypothyroidism were similar to IPAH with euthyroid function except for lower right atrial pressure (6 versus 8 mmHg; p=0.009). The long-term survival of patients with clinical thyroid dysfunction was better than IPAH without thyroid dysfunction, while that of those with subclinical diseases was comparable to the latter, even after adjusting for baseline haemodynamics and treatment. IPAH patients with clinical hyper- and hypothyroidism had better haemodynamics and survival than those without thyroid dysfunction, while patients with subclinical hypothyroidism had similar haemodynamics and survival profiles to the latter.