Breakthrough SARS-COV-2 infection induces broad anti-viral T cell immunity.

Vaccines have curtailed the devastation wrought by COVID-19. Nevertheless, emerging variants result in a high incidence of breakthrough infections. Here we assess the impact of vaccination and breakthrough infection on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) T cell immunity. We demonstrate that COVID-19 vaccination induces robust spike-specific T cell responses that, within the CD4+ compartment, display comparable IFN-γ responses to SARS-CoV-2 infection without vaccination. Vaccine-induced CD8+ IFN-γ responses however, were significantly greater than those primed by SARS-CoV-2 infection alone. This increased responsiveness is associated with induction of novel HLA-restricted CD8+ T cell epitopes not primed by infection alone (without vaccination). Despite these augmented responses, breakthrough infection still induced de novo T cell responses against additional SARS-CoV-2 CD8+ epitopes that display HLA-associated immunodominance hierarchies consistent with those in unvaccinated COVID-19 convalescent individuals. This study demonstrates the unique modulation of anti-viral T cell responses against multiple viral antigens following consecutive yet distinct priming events in COVID-19 vaccination and breakthrough infection.

Exaggerated myocardial torsion may contribute to dynamic left ventricular outflow tract obstruction in hypertrophic cardiomyopathy.

Aims: Dynamic left ventricular (LV) outflow tract obstruction (LVOTO) is associated with symptoms and increased risk of developing heart failure in hypertrophic cardiomyopathy (HCM). The association of LVOTO and LV twist mechanics has not been well studied in HCM. The aim of the study was to compare the pattern of LV twist in patients with HCM associated with asymmetrical septal hypertrophy with and without LVOTO. Methods and results: Echocardiography (including speckle tracking) was performed in 212 patients with HCM, divided according to the absence (n = 130) or presence (n = 82) of LVOTO (defined as peak pressure gradient ≥30 mmHg either at rest and/or with Valsalva manoeuvre). Patients with LVOTO were older, had smaller LV dimensions, a higher LV ejection fraction (LVEF), a longer anterior mitral valve leaflet length, and a higher early transmitral pulsed wave to septal tissue Doppler velocity ratio (E/E'). A univariate analysis showed that peak twist was significantly higher in patients with LVOTO compared with patients without LVOTO (19.7 ± 7.3 vs. 15.7 ± 6.0, P = 0.00015). Peak twist was similarly enhanced in patients with LVOTO, manifesting only during Valsalva (19.2 ± 5.6, P = 0.007) and patients with resting LVOTO (19.9 ± 8.0, P = 0.00004) compared with patients without LVOTO (15.7 ± 6.0). A stepwise forward logistic regression analysis showed that LVEF, LV end-systolic dimension indexed to body surface area, anterior mitral valve leaflet length, E/E', and peak twist were all independently associated with LVOTO. Conclusion: This study demonstrates that increased peak LV twist is independently associated with LVOTO in patients with HCM. Peak twist was similarly exaggerated in patients with only latent LVOTO, suggesting that it may play a contributory role to LVOTO in HCM.

Critical illness associated new onset atrial fibrillation: subsequent atrial fibrillation diagnoses and other adverse outcomes.

AIMS: Amongst patients with critical illness associated new onset AF (CI-NOAF), the risk of subsequent atrial fibrillation (AF) diagnoses and other adverse outcomes is unknown, and the role for long-term anticoagulation is unclear. This study sought to determine the factors associated with subsequent AF diagnoses and other adverse outcomes in this cohort. METHODS AND RESULTS: Admissions to a tertiary general intensive care unit (ICU) between December 2015 and September 2018 were screened for AF episodes through hourly analysis of continuous ECG monitoring. Patients with a prior history of AF were excluded. AF burden was defined as the percentage of monitored ICU hours in AF. The primary endpoint was subsequent AF diagnoses, as collated from the statewide electronic medical records. Secondary endpoints included mortality, embolic events, MACE and subsequent anticoagulation. RESULTS: Of 7030 admissions with 509 303 h of monitoring data, 309 patients with CI-NOAF were identified, and 235 survived to discharge. Subsequent AF diagnoses were identified in 75 (31.9%) patients after a median of 413 days. Increased AF burden had the strongest independent association with AF recurrence (OR = 15.03, P = 0.002), followed by increased left atrial area (OR = 1.12, P = 0.01). Only 128 (54.5%) patients had their AF diagnosis acknowledged at ICU discharge, and 50 (21.3%) received anticoagulation at hospital discharge. CONCLUSION: CI-NOAF is often under-recognized, and subsequent AF diagnoses are common post-discharge. AF burden during ICU admission has a strong independent association with subsequent AF diagnoses. Left atrial size is also independently associated with subsequent AF.

Towards Label-free detection of viral disease agents through their cell surface proteins: Rapid screening SARS-CoV-2 in biological specimens.

Current methods for the screening of viral infections in clinical settings, such as reverse transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), are expensive, time-consuming, require trained personnel and sophisticated instruments. Therefore, novel sensors that can save time and cost are required specially in remote areas and developing countries that may lack the advanced scientific infrastructure for this task. In this work, we present a sensitive, and highly specific biosensing approach for the detection of harmful viruses that have cysteine residues within the structure of their cell surface proteins. We utilized new method for the rapid screening of SARS-CoV-2 virus in biological fluids through its S1 protein by surface enhanced Raman spectroscopy (SERS). The protein is captured from aqueous solutions and biological specimens using a target-specific extractor substrate. The structure of the purified protein is then modified to convert it into a bio-thiol by breaking the disulfide bonds and freeing up the sulfhydryl (SH) groups of the cysteine residues. The formed biothiol chemisorbs favourably onto a highly sensitive plasmonic sensor and probed by a handheld Raman device in few seconds. The new method was used to screen the S1 protein in aqueous medium, spiked human blood plasma, mucus, and saliva samples down to 150 fg/L. The label-free SERS biosensing method has strong potential for the fingerprint identification many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method can be applied at points of care (POC) in remote areas and developing countries lacking sophisticated scientific infrastructure.

A paper-based optical sensor for the screening of viruses through the cysteine residues of their surface proteins: A proof of concept on the detection of coronavirus infection.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a serious threat to human health. Current methods such as reverse transcription polymerase chain reaction (qRT-PCR) are complex, expensive, and time-consuming. Rapid, and simple screening methods for the detection of SARS-CoV-2 are critically required to fight the current pandemic. In this work we present a proof of concept for, a simple optical sensing method for the screening of SARS-CoV-2 through its spike protein subunit S1. The method utilizes a target-specific extractor chip to bind the protein from the biological specimens. The disulfide bonds of the protein are then reduced into a biothiol with sulfhydryl (SH) groups that react with a blue-colored benzothiazole azo dye-Hg complex (BAN-Hg) and causes the spontaneous change of its blue color to pink which is observable by the naked eye. A linear relationship between the intensity of the pink color and the logarithm of reduced S1 protein concentration was found within the working range 130 ng.mL-1-1.3 pg mL-1. The lowest limit of detection (LOD) of the assay was 130 fg mL-1. A paper based optical sensor was fabricated by loading the BAN-Hg sensor onto filter paper and used to screen the S1 protein in spiked saliva and patients' nasopharyngeal swabs. The results obtained by the paper sensor corroborated with those obtained by qRT-PCR. The new paper-based sensing method can be extended to the screening of many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method has strong potential for screening viruses at pathology labs and in remote areas that lacks advanced scientific infrastructure. Further clinical studies are warranted to validate the new sensing method.

Composite Echocardiographic Score to Predict Long-Term Survival Following Myocardial Infarction.

BACKGROUND: Whilst the left ventricular ejection fraction (LVEF) remains the primary echocardiographic measure widely utilised for risk stratification following myocardial infarction (MI), it has a number of well recognised limitations. The aim of this study was to compare the prognostic utility of a composite echocardiographic score (EchoScore) composed of prognostically validated measures of left-ventricular (LV) size, geometry and function, to the utility of LVEF alone, for predicting survival following MI. METHODS: Retrospective data on 394 consecutive patients with a first-ever MI were included. Comprehensive echocardiography was performed within 24 hours of admission for all patients. EchoScore consisted of LVEF<50%, left atrial volume index>34 mL/m2, average E/e >14, E/A ratio>2, abnormal LV mass index, and abnormal LV end-systolic volume index. A single point was allocated for each measure to derive a score out of 6. The primary outcome measure was all-cause mortality. RESULTS: At a median follow-up of 24 months there were 33 deaths. On Kaplan-Meier analysis, a high EchoScore (>3) displayed significant association with all-cause mortality (log-rank χ2=74.48 p<0.001), and was a better predictor than LVEF<35% (log-rank χ2=17.01 p<0.001). On Cox proportional-hazards multivariate analysis incorporating significant clinical and echocardiographic predictors, a high EchoScore was the strongest independent predictor of all-cause mortality (HR 6.44 95%CI 2.94-14.01 p<0.001), and the addition of EchoScore resulted in greater increment in model power compared to addition of LVEF (model χ2 56.29 vs 44.71 p<0.001, Harrell's C values 0.83 vs 0.79). CONCLUSIONS: A composite echocardiographic score composed of prognostically validated measures of LV size, geometry, and function is superior to LVEF alone for predicting survival following MI.

Accuracy of Cardiac Nurse Acquired and Measured Three-Dimensional Echocardiographic Left Ventricular Ejection Fraction: Comparison to Echosonographer.

BACKGROUND: Three-dimensional echocardiography (3D-Echo) performed by novice health care staff to measure left ventricular ejection fraction (LVEF) could allow cost-effective screening and monitoring for left ventricular systolic dysfunction (LVSD) prior to the development of heart failure. The aim of this study was to determine feasibility and accuracy of cardiac nurses (after completing focussed training) independently acquiring 3D-Echo images, and measuring LVEF using semi-automated software when compared to an echosonographer. METHODS: One echosonographer and three cardiac nurses acquired 3D-Echo images on 73 patients (62 ± 16 years, 62% male) with good image quality, and subsequently measured LVEF using a semi-automated algorithm. RESULTS: Overall feasibility was 89% with the three nurses successfully acquiring 3D-Echo images suitable for LVEF assessment in 65 of the 73 patients. High accuracy (r = 0.82; p < 0.0001) with minimal bias (+0.1, -10.6 to +10.8 limits of agreement; p = 0.91) was observed comparing the nurses to the echosonographer for measuring LVEF. Individual nurses demonstrated high feasibility (86%-92%), accuracy (r = 0.83-0.87; all p < 0.0001) and intra-observer reproducibility (r = 0.96-0.97; all p < 0.0001), with good inter-observer consistency in accuracy compared to the echosonographer (one-way analysis of variance p = 0.559). CONCLUSIONS: We have demonstrated that, following a focussed training protocol, it was feasible for cardiac nurses to acquire 3D-Echo images of sufficient image quality to allow measurement of LVEF using a semi-automated algorithm, with comparable accuracy and intra-observer variability to an expert echosonographer. This could potentially allow the broader application of echocardiography to screen for LVSD in high-risk cohorts.

Hemodynamic Validation of the E/e' Ratio as a Measure of Left Ventricular Filling Pressure in Patients With Non-ST Elevation Myocardial Infarction.

The E/e' ratio has an established role in the assessment of left ventricular filling pressure (LVFP) in stable patients, but its accuracy in acute myocardial ischemia is less well established. The aim of this study was to validate the relation between the E/e' ratio and invasively measured LVFP in patients with non-ST elevation myocardial infarction (NSTEMI). A total of 120 unselected patients with NSTEMI underwent cardiac catheterization with measurement of left ventricular end-diastolic pressure (LVEDP; elevated ≥15 mm Hg) and Doppler echocardiography with either simultaneous (n = 30) or same-day (n = 90) measurement of E/e'. Patients were aged 64.1 ± 11.8 years, 72% were male and mean left ventricular ejection fraction was 48.0 ± 20.9%. Septal, lateral, and average E/e' ratios all showed a significant correlation with LVEDP (Pearson's r: 0.42, 0.43, 0.48, respectively [all p <0.001]). Receiver operating characteristics curves showed an area under the curve of 0.72, 0.72, and 0.75 (all p <0.001) for septal, lateral, and average E/e', respectively. The sensitivity, specificity, positive (PPV), and negative (NPV) predictive values for the guideline-recommended threshold of average E/e' >14 for elevated LVEDP was 27%, 93%, 79%, and 44%, respectively. Utilizing lower E/e' boundaries of 6, 7, and 8 for lateral, average, and medial E/e', respectively, improved the NPV to ≥80% for each parameter. In conclusion, the E/e' ratio is a robust measure of LVFP during acute NSTEMI using upper and lower thresholds to achieve a high PPV and NPV, respectively, with the use of adjunctive guideline-recommend measures required in patients with nonconclusive E/e'.

Salivary Protein Panel to Diagnose Systolic Heart Failure.

Screening for systolic heart failure (SHF) has been problematic. Heart failure management guidelines suggest screening for structural heart disease and SHF prevention strategies should be a top priority. We developed a multi-protein biomarker panel using saliva as a diagnostic medium to discriminate SHF patients and healthy controls. We collected saliva samples from healthy controls (n = 88) and from SHF patients (n = 100). We developed enzyme linked immunosorbent assays to quantify three specific proteins/peptide (Kallikrein-1, Protein S100-A7, and Cathelicidin antimicrobial peptide) in saliva samples. The analytical and clinical performances and predictive value of the proteins were evaluated. The analytical performances of the immunoassays were all within acceptable analytical ranges. The multi-protein panel was able to significantly (p < 0.001) discriminate saliva samples collected from patients with SHF from controls. The multi-protein panel demonstrated good performance with an overall diagnostic accuracy of 81.6% (sensitivity of 79.2% and specificity of 85.7%) when distinguishing SHF patients from healthy individuals. In conclusion, we have developed immunoassays to measure the salivary concentrations of three proteins combined as a panel to accurately distinguish SHF patients from healthy controls. While this requires confirmation in larger cohorts, our findings suggest that this three-protein panel has the potential to be used as a biomarker for early detection of SHF.

Echocardiographic predictors of all-cause mortality in patients with left ventricular ejection fraction >35%: Value of guideline based assessment of diastolic dysfunction.

BACKGROUND: Recent data suggests that the majority of cardiac deaths in patients with heart failure occur in patients with a left ventricular ejection fraction (LVEF) >35%. This study sought to determine the value of guideline based assessment of diastolic dysfunction in predicting all-cause mortality in patients with a first-ever myocardial infarction (MI) with an LVEF >35%. METHODS: A retrospective single centre study involving 383 patients with a first-ever MI (STEMI or NSTEMI) with LVEF >35% was performed. Clinical, angiographic and echocardiographic data were obtained from prospectively maintained institutional databases. Outcomes data were obtained from national death registry. Echocardiography was performed early post-admission for all patients. Significant diastolic dysfunction (DD) was defined was grade 2/3 diastolic dysfunction according to current American Society of Echocardiography/European Association of Cardiovascular Imaging guidelines. RESULTS: At a median follow up of 2 years, there were 32 deaths. On Cox proportional hazards multivariate analysis incorporating significant clinical variables (age, chronic kidney disease and extent of coronary artery disease), significant DD (HR 2.57, 95%CI 1.16-5.68, p = 0.020) and left ventricular end-diastolic volume index (HR 1.03, 1.04-1.07, p = 0.021) were the only independent echocardiographic predictors of all-cause mortality. Intermodel comparisons using model χ2 and Harrel's-C confirmed incremental value of DD. In the subgroup with LVEF 36-55% (n = 176), significant DD was the only independent echocardiographic predictor (HR 3.56, 95%CI 2.46-9.09, p = 0.006). CONCLUSIONS: The presence of significant DD identifies patients with LVEF >35% following MI who are at a higher risk of all-cause mortality, and who may benefit from further risk stratification and treatment.

Relation of Left Atrial Volumes in Patients With Myocardial Infarction to Left Ventricular Filling Pressures and Outcomes.

The inter-relationships between minimal and maximal left atrial volume index (LAVI), left ventricular filling pressures and survival have not been well studied. This study aimed to compare LAVImin with LAVImax with respect to (1) relative prognostic value, and (2) correlation with left ventricular end-diastolic pressures (LVEDP), in patients with myocardial infarction (MI). A retrospective study involving consecutive patients with a first-ever MI (n = 419) was undertaken. LAVIs were determined using Simpson's biplane method from 2D echocardiography performed the day after admission. LAVmin ≥ 18 mls/m2 and LAVImax ≥ 34 mls/m2 were considered enlarged. The primary end point was composite major adverse cardiovascular events (MACE) (death/MI/heart failure). Correlation between LVEDP and LAVI was assessed in 120 patients who underwent echocardiography and cardiac catheterization either simultaneously (n = 30) or same-day (n = 90). At a median follow-up of 24 months, there were 61 MACE events. On Cox proportional hazards multivariate analysis incorporating significant clinical predictors and LVEF, whereas both LAVImin ≥ 18 mls/m2 (hazard ratio 3.15 [95% confidence interval 1.70 to 5.54], p <0.001) and LAVImax ≥ 34 mls/m2 (hazard ratio 1.79 [95% confidence interval 1.02 to 3.14], p = 0.041) were independent predictors of MACE, LAVImin showed a stronger association. Intermodel comparisons of the model chi-square and Harrell's C-statistic confirmed better prognostication with LAVImin. In the invasive cohort, because LAVImin and LAVImax had a similar correlation with LVEDP ≥ 15 mm Hg (r = 0.41 [p <0.001] vs r = 0.42 [p <0.001]), LAVmin ≥ 18 mls/m2 had a greater sensitivity for LVEDP ≥ 15 mm Hg than LAVImax ≥ 34 mls/m2 (sensitivity 59.4% vs 34.4%). In conclusion, utilizing thresholds of ≥18 and ≥34 mls/m2, respectively, LAVImin was a better predictor of survival than LAVImax, the pathophysiologic basis of which relates to a better sensitivity for elevated left ventricular filling pressures with LAVImin at these thresholds. There may be incremental clinical value in measuring LAVImin alongside LAVImax.

Diastolic Dysfunction Assessed Using Contemporary Guidelines and Prognosis Following Myocardial Infarction.

BACKGROUND: Recent American Society of Echocardiography and European Association of Cardiovascular Imaging guidelines for the assessment of diastolic dysfunction (DD) recommend a simplified approach with four key variables incorporated into a novel diagnostic algorithm. The aim of this study was to assess the prognostic value of significant DD assessed using the algorithm recommended in the 2016 American Society of Echocardiography and European Association of Cardiovascular Imaging guidelines (DD2016) in comparison with the prognostic value of significant DD assessed using the 2009 guidelines (DD2009) as well as the individual parameters incorporated in the 2016 algorithm. METHODS: Retrospective data on 419 consecutive patients with first ever myocardial infarction were included. Doppler echocardiography was performed within 24 hours of admission in all patients. Significant DD was defined as grade 2 or 3 DD. The primary outcome measure was composite major adverse cardiovascular events (MACEs), comprising death, myocardial infarction, and heart failure. RESULTS: At a median follow-up of 24 months, there were 61 MACEs. On Kaplan-Meier analysis, DD2016 showed a better association with MACEs than DD2009 (log-rank χ2 = 21.01 [P < .001] vs 13.13 [P = .001]). On Cox proportional-hazards multivariate analysis incorporating significant clinical predictors and left ventricular ejection fraction, DD2016 (hazard ratio, 2.22; 95% CI, 1.25-3.98; P = .007) was the strongest independent predictor of MACEs, whereas DD2009 (hazard ratio, 1.63; 95% CI, 0.95-2.80; P = .074) was not a significant predictor. Of the four key diastolic parameters, only left atrial volume index was independently associated with MACEs (hazard ratio, 1.79; 95% CI, 1.02-3.14; P = .041) when included in a Cox proportional-hazards multivariate model incorporating significant clinical predictors and left ventricular ejection fraction, although the association was weaker than DD2016. Intermodel comparisons with model χ2 and Harrell's C statistic were satisfactory for DD2016. CONCLUSIONS: Significant DD assessed using the 2016 American Society of Echocardiography and European Association of Cardiovascular Imaging guidelines is a robust independent predictor of clinical outcomes following myocardial infarction and compares favorably with DD2009 as well as the individual parameters incorporated in the novel 2016 algorithm.

Evaluation of training nurses to perform semi-automated three-dimensional left ventricular ejection fraction using a customised workstation-based training protocol.

AIMS AND OBJECTIVES: We aimed to determine the feasibility of training cardiac nurses to evaluate left ventricular function utilising a semi-automated, workstation-based protocol on three dimensional echocardiography images. BACKGROUND: Assessment of left ventricular function by nurses is an attractive concept. Recent developments in three dimensional echocardiography coupled with border detection assistance have reduced inter- and intra-observer variability and analysis time. This could allow abbreviated training of nurses to assess cardiac function. DESIGN: A comparative, diagnostic accuracy study evaluating left ventricular ejection fraction assessment utilising a semi-automated, workstation-based protocol performed by echocardiography-naïve nurses on previously acquired three dimensional echocardiography images. METHODS: Nine cardiac nurses underwent two brief lectures about cardiac anatomy, physiology and three dimensional left ventricular ejection fraction assessment, before a hands-on demonstration in 20 cases. We then selected 50 cases from our three dimensional echocardiography library based on optimal image quality with a broad range of left ventricular ejection fractions, which was quantified by two experienced sonographers and the average used as the comparator for the nurses. Nurses independently measured three dimensional left ventricular ejection fraction using the Auto lvq package with semi-automated border detection. RESULTS: The left ventricular ejection fraction range was 25-72% (70% with a left ventricular ejection fraction <55%). All nurses showed excellent agreement with the sonographers. Minimal intra-observer variability was noted on both short-term (same day) and long-term (>2 weeks later) retest. CONCLUSIONS: It is feasible to train nurses to measure left ventricular ejection fraction utilising a semi-automated, workstation-based protocol on previously acquired three dimensional echocardiography images. Further study is needed to determine the feasibility of training nurses to acquire three dimensional echocardiography images on real-world patients to measure left ventricular ejection fraction. RELEVANCE TO CLINICAL PRACTICE: Nurse-performed evaluation of left ventricular function could facilitate the broader application of echocardiography to allow cost-effective screening and monitoring for left ventricular dysfunction in high-risk populations.