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Recent studies have suggested that textural characteristics of the intima-media complex (IMC) may be more useful than the intima-media thickness (IMT) in evaluating cardiovascular risk. The primary aim of our study was to investigate the association between texture features of the common carotid IMC and prevalent clinical cardiovascular disease (CVD). The secondary aim was to determine whether IMT and IMC texture features vary between the left and right carotid arteries. The study was performed on 2208 longitudinal-section ultrasound images of the left (L) and right (R) common carotid artery (CCA), acquired from 569 men and 535 women out of which 125 had clinical CVD. L and R sides of the IMC were intensity normalized and despeckled. The IMC was semiautomatically delineated for all images using a semiautomated segmentation system, and 61 different texture features were extracted. The corresponding IMT semiautomated measurements (mean±SD) of the L and R sides were 0.73±0.21 mm/0.69±0.19 mm for the normal population and 0.83±0.17 mm/0.79±0.18 mm for those with CVD. IMC texture features did not differ between the right- and left-hand sides. Several texture features were independent predictors of the presence of CVD. The multivariate logistic regression analysis combining age, IMT, and texture features produced a receiver operating characteristic curve with an area under the curve of 89%. A correct classification rate of 77% for separating the normal subject (NOR) versus CVD subjects was achieved using the support vector machine classifier with a combination of clinical features, IMT, and extracted texture features. Texture features provide additional information on the presence of clinical CVD, which is over and above that provided by conventional risk factors or IMT alone. The value of IMC texture features in the prediction of future cardiovascular events should be tested in prospective studies.

We previously showed that use of portable noninvasive ventilation (pNIV) during recovery periods within intermittent exercise improved breathlessness and exercise tolerance in patients with COPD compared with pursed-lip breathing (PLB). However, in a minority of patients recovery from dynamic hyperinflation (DH) was better with PLB, based on inspiratory capacity. We further explored this using Optoelectronic Plethysmography to assess total and compartmental thoracoabdominal volumes. Fourteen patients with COPD (means ± SD) (FEV1: 55% ± 22% predicted) underwent, in a balanced order sequence, two intermittent exercise protocols on the cycle ergometer consisting of five repeated 2-min exercise bouts at 80% peak capacity, separated by 2-min recovery periods, with application of pNIV or PLB in the 5 min of recovery. Our findings identified seven patients showing recovery in DH with pNIV (DH responders) whereas seven showed similar or better recovery in DH with PLB. When pNIV was applied, DH responders compared with DH nonresponders exhibited greater tidal volume (by 0.8 ± 0.3 L, P = 0.015), inspiratory flow rate (by 0.6 ± 0.5 L/s, P = 0.049), prolonged expiratory time (by 0.6 ± 0.5 s, P = 0.006), and duty cycle (by 0.7 ± 0.6 s, P = 0.007). DH responders showed a reduction in end-expiratory thoracoabdominal DH (by 265 ± 633 mL) predominantly driven by reduction in the abdominal compartment (by 210 ± 494 mL); this effectively offset end-inspiratory rib-cage DH. Compared with DH nonresponders, DH responders had significantly greater body mass index (BMI) by 8.4 ± 3.2 kg/m2, P = 0.022 and tended toward less severe resting hyperinflation by 0.3 ± 0.3 L. Patients with COPD who mitigate end-expiratory rib-cage DH by expiratory abdominal muscle recruitment benefit from pNIV application.\ud \ud NEW & NOTEWORTHY Compared with the pursed-lip breathing technique, acute application of portable noninvasive ventilation during recovery from intermittent exercise improved end-expiratory thoracoabdominal dynamic hyperinflation (DH) in 50% of patients with COPD (DH responders). DH responders, compared with DH nonresponders, exhibited a reduction in end-expiratory thoracoabdominal DH predominantly driven by the abdominal compartment that effectively offset end-expiratory rib cage DH. The essential difference between DH responders and DH nonresponders was, therefore, in the behavior of the abdomen.

Mutations of calreticulin (CALR) are the second most prevalent driver mutations in essential thrombocythemia and primary myelofibrosis. To identify potential targeted therapies for CALR mutated myeloproliferative neoplasms, we searched for small molecules that selectively inhibit the growth of CALR mutated cells using high-throughput drug screening. We investigated 89 172 compounds using isogenic cell lines carrying CALR mutations and identified synthetic lethality with compounds targeting the ATR-CHK1 pathway. The selective inhibitory effect of these compounds was validated in a co-culture assay of CALR mutated and wild-type cells. Of the tested compounds, CHK1 inhibitors potently depleted CALR mutated cells, allowing wild-type cell dominance in the co-culture over time. Neither CALR deficient cells nor JAK2V617F mutated cells showed hypersensitivity to ATR-CHK1 inhibition, thus suggesting specificity for the oncogenic activation by the mutant CALR. CHK1 inhibitors induced replication stress in CALR mutated cells revealed by elevated pan-nuclear staining for γH2AX and hyperphosphorylation of RPA2. This was accompanied by S-phase cell cycle arrest due to incomplete DNA replication. Transcriptomic and phosphoproteomic analyses revealed a replication stress signature caused by oncogenic CALR, suggesting an intrinsic vulnerability to CHK1 perturbation. This study reveals the ATR-CHK1 pathway as a potential therapeutic target in CALR mutated hematopoietic cells.

10 p.-6 fig., Establishing causal links between bacterial metabolites and human intestinal disease is a significant challenge. This study reveals the molecular basis of antibiotic-associated hemorrhagic colitis (AAHC) caused by intestinal resident Klebsiella oxytoca Colitogenic strains produce the nonribosomal peptides tilivalline and tilimycin. Here, we verify that these enterotoxins are present in the human intestine during active colitis and determine their concentrations in a murine disease model. Although both toxins share a pyrrolobenzodiazepine structure, they have distinct molecular targets. Tilimycin acts as a genotoxin. Its interaction with DNA activates damage repair mechanisms in cultured cells and causes DNA strand breakage and an increased lesion burden in cecal enterocytes of colonized mice. In contrast, tilivalline binds tubulin and stabilizes microtubules leading to mitotic arrest. To our knowledge, this activity is unique for microbiota-derived metabolites of the human intestine. The capacity of both toxins to induce apoptosis in intestinal epithelial cells-a hallmark feature of AAHC-by independent modes of action, strengthens our proposal that these metabolites act collectively in the pathogenicity of colitis., We acknowledge networking contributions from European Cooperation in Science and Technology Action CM1407, “Challenging Organic Syntheses Inspired by Nature—From Natural Products Chemistry to Drug Discovery.” Research was funded by the Austrian Science Fund W901 Doktoratskolleg Molecular Enzymology (to E.L.Z., S.S., W.K., and R.B.), BioTechMed-Graz Secretome Flagship (to S.S., E.L.Z., G.G., C.H., and R.B.), and Grant BFU2016-75319-R (Agencia Española de Investigacion/Fondo Europeo de desarrollo regional, Unión Europea) (to J.F.D.) from Ministerio de Economica y Competitividad.

Measurements of ultrasound diaphragmatic motion, amplitude, force, and velocity of contraction may provide important and essential information about diaphragmatic fatigue, weakness, or paralysis. In this paper, we propose and evaluate a semi-automated analysis system for measuring the diaphragmatic motion and estimating the maximum relaxation rate (MRR_SAUS) from ultrasound M-mode images of the diaphragmatic muscle. The system was evaluated on 27 M-mode ultrasound images of the diaphragmatic muscle [20 with no resistance (NRES) and 7 with resistance (RES)]. We computed semi-automated ultrasound MRR measurements on all NRES/RES images, using the proposed system (MRR_SAUS = 3.94 ± 0.91/4.98 ± 1.98 [1/s]), and compared them with the manual measurements made by a clinical expert (MRR_MUS = 2.36 ± 1.19/5.8 ± 2.1 [1/s],) and those made by a reference manual method (MRR_MB = 3.93 ± 0.89/3.73 ± 0.52 [1/sec], performed manually with the Biopac system. MRR_SAUS and MRR_MB measurements were not statistically significantly different for NRES and RES subjects but were significantly different with the MRR-MUS measurements made by the clinical expert. It is anticipated that the proposed system might be used in the future in the clinical practice in the assessment and follow up of patients with diaphragmatic weakness or paralysis. It may thus potentially help to understand post-operative pulmonary dysfunction or weaning failure from mechanical ventilation. Further validation and additional experimentation in a larger sample of images and different patient groups is required for further validating the proposed system., Illustration of the steps followed for the semi-automated measurement of the diaphragmatic MRR_SAUS index. (a) Initial ultrasound M-mode image of the diaphragmatic motion with the selected rectangular ROI and the calibration points placed manually by the user of the system for ten breathing cycles. The ultrasound M-mode image was acquired from a male subject at the age of 45 without the use of resistance during breathing (NRES). (b) Diaphragmatic ultrasound motion curve after image normalization and despeckle filtering. (c) Binary image from (c). (d) Extraction of the upper and lower boundaries of the diaphragmatic motion curve. (e) Final upper and lower boundaries overlaid on the grayscale diaphragmatic ultrasound image. (f) Upper smoothed graph of the diaphragmatic displacement for the first four breaths with absolute maximum [4.59 mm, 12.72 mm] and minimum [3.24 mm, 4.79 mm] values and the steepest slope [23.47 mm/sec], at the coordinates [3.51 sec, 7.92 mm] and [3.54 sec, 8.52 mm]. Absolute maximum and minimum distance (diaphragmatic displacement): 12.72-4.79

In the course of a human brain acquisition, which is acquired by a magnetic resonance imager (MRI), two-dimensional (2D) slices of the brain are captured. These have to be aligned and reconstructed to a three-dimensional (3D) volume, which will better assist the doctor in following up the development of the disease. In this study, a 3D reconstruction integrated system for MRI brain multiple sclerosis (MS) lesion visualization is proposed. Brain MRI images from 5 MS subjects were acquired at four diffident consecutive time points (TP1-TP4) with an interval of 6–12 months. MS lesions were manually segmented by an expert neurologist and semi-automatically by a system and reconstructed in a brain volume. The proposed system assists the doctor in following up the MS disease progression and provides support to better manage the disease. The proposed system includes a 5-stage investigation (pre-processing, lesion segmentation, 3D reconstruction, volume estimation and method evaluation), as well as a module for the quantitative evaluation of the method. Twenty MRI images of the brain were used to evaluate the proposed system. Results show that the 3D reconstruction method proposed in this work, can be used to differentiate brain tissues and recognize MS lesions by providing improved 3D visualization. These preliminary results provide evidence that the proposed system could be applied in the future in clinical practice given that it is further evaluated on more subjects.

Introduction Many patients with epistaxis are seen in the A&E department. A significant number are discharged without need for inpatient admission due to minor bleeds. Many are given limited information on management of further bleeds and even fewer are advised on the prevention of bleeds. Aim To evaluate the advice given to patients with epistaxis who are seen and discharged from A+E, to instigate an improvement in advice and to examine the subsequent effect on re-attendance rates Method Epistaxis patients discharged from A+E were audited over a 6-week period looking at advice given on discharge. A questionnaire was distributed to all A&E and ENT doctors probing current practice. A presentation on the management and prevention of epistaxis and the appropriate verbal/written advice to give was delivered and epistaxis advice sheets were made available. The practice regarding advice and the re-attendance rates were re-audited. Results 53 patients presented to the A&E department with epistaxis. 31 patients presenting with epistaxis were seen and discharged by A&E doctors. The remaining seen by ENT. Only 9 patients received verbal advice and 1 received written advice. There were 13 re-attenders. After re-audit showed a significant increase in the proportion of patients receiving advice by A&E doctors. Verbal and written advice increased from 18% to 61% and 1% to 60% respectively. The number of re-attenders was reduced to 13 to 6.

Breast cancer brain metastasis (BCBM) still remains a major clinical challenge. Current systemic treatments are often inadequate while diagnosis involves time-consuming series of neuro-imaging acquisitions and dangerous invasive biopsies. Automated image analysis systems for the identification, prediction and follow up of BCBM are therefore required. This review discusses the advancements in the automated MRI brain metastasis (BM) image analysis using radiomic features based classification. Seven BM segmentation studies, and three BCBM identification studies were considered eligible. The latter studies were based on either manual or semi-automated segmentation methods. Almost every fully automated BM segmentation method presented in the literature, reported a maximum dice similarity score (DSC) of 84%, but they resulted in a poor BM segmentation for brain areas less than 5 mm (0.06 ml). The multi-class prediction of BCBM approach, which is more representative for clinical applicability, is based on imaging features and resulted in an area under the curve (AUC) of 60%. Therefore, the need still exists for the development of automated image analysis methods for the identification, follow up and prediction of BCBM. The potential clinical usage of above methods entails further multi-center studies with comprehensive clinical data and multi-class modeling with vast and varying primary and metastatic brain tumors.

Worry and rumination, two forms of repetitive negative thinking (RNT), are prevalent in generalized anxiety disorder (GAD) and depression. Cognitive processing biases, especially the tendency to draw negative conclusions from ambiguous information (interpretation bias), may maintain worry and rumination. Yet the relationship between interpretation bias and both forms of RNT has not been explored in clinical versus nonclinical samples. In this cross-sectional study, participants with GAD ( n = 72), depression ( n = 79), or neither disorder ( n = 71) completed two tasks assessing interpretation bias, measures of worry and rumination, and reported negative thought intrusions during a behavioral task. Interpretation bias was associated with higher levels of worry, rumination, and negative thought intrusions. Both clinical groups generated significantly more negative interpretations than healthy comparison participants. These findings link interpretation bias to worry and rumination and establish the need for research investigating the causal role of interpretation bias in maintaining RNT.

The deterioration of sleep quality (SQ) can be a manifestation of mental health issues and lack of Self-Esteem (SE), especially in men, who express their mental discomfort in ways that can be distinct from those of women. The present study sought to examine whether depressive symptoms and trait anxiety mediate the association between SE and SQ in male university students. 142 university students were evaluated using the Rosenberg Self-Esteem scale, the Beck Depression Inventory, the Spielberger Trait Anxiety Inventory, and the Pittsburg Quality Sleep Index. The analysis was based on the Baron and Kenny four steps mediation model, and explored the potential mediator role of depression and trait anxiety through a series of simple and multiple linear regression analyses, followed by a Sobel test to corroborate the model's power. Our results showed that the association between SE and SQ is partially mediated by depression and trait anxiety. Such findings may indicate that both depression and trait anxiety contribute to the underlying mechanism through which SE influences SQ in male university students. Clinical and educational implications are discussed in the light of such results, proposing interventions to ameliorate the male students' emotional life and sleep quality.

Chronic tendinopathy represents a growing burden to healthcare services in an active and ageing global population. The ability to identify, isolate and interrogate, in vitro, key pathogenic and reparative tendon cell populations is essential to developing precision therapies and implantable materials.Human hamstring tendon cells were cultured for 8 days on either tissue culture plastic or aligned electrospun fibres made of polydioxanone (absorbable polymer). Combined single cell surface proteomics and unbiased single cell transcriptomics (CITE-Seq) revealed six discrete cell clusters, four of which shared key gene expression determinants with ex vivo human cell clusters. These were PTX3_PAPPA, POST_SCX, DCN_LUM and ITGA7_NES cell clusters. Surface proteomics found that PTX3_PAPPA cells were CD10+CD26+CD54+. ITGA7_NES cells were CD146+, and POSTN_SCX cells were CD90+CD95+CD10+.Three clusters preferentially survived and proliferated on the aligned electrospun fibres; DCN_LUM, POSTN_SCX, and PTX3_PAPPA. They maintained high expression of tendon matrix associated genes, includingCOL1A1, COL1A2, COL3A1, ELN, FBLN1, and up-regulated genesets enriched for TNF-Ɣsignalling via NFκB, IFN-Ɣ signalling and IL-6/ STAT3 signalling. When cells were pre-selected based on surface protein markers, a similar up-regulation of pro-inflammatory signalling pathways was observed, particularly inPTX3gene expressing CD10+CD26+CD54+ cells, with increased expression of genes associated with TNF-αsignalling and IFN-γ signalling.Discrete human tendon cell sub populations persist in vitro culture and can be recognised by specific gene and surface protein signatures. Aligned PDO fibres promote the survival of three clusters, including pro-inflammatoryPTX3expressing CD10+CD26+CD54+ cells found in chronic tendon disease.

Carcinogenic effects of Aspergillus spp. have been well established and generally attributed to a variety of mycotoxin productions, particularly aflatoxins. It is known that most carcinogenic mycotoxins, with the exception of fumonisins, are genotoxic and mutagenic, causing chromosomal aberrations, micronuclei, DNA single-strand breaks, sister chromatid exchange, unscheduled DNA synthesis etc. Some Aspergillus spp. are infected with mycoviruses which can result in loss of aflatoxin production. The effects of mycovirus containing Aspergillus on human health have not been fully evaluated. Recent studies in patients with acute lymphoblastic leukemia, in full remission, have revealed the existence of antibody to the products of a certain Aspergillus flavus isolate which harbored an unknown mycovirus. Exposure of blood mononuclear cells from these patients, but not controls, to the products of this organism had reproduced cell surface phenotypes and genetic markers, characteristic of acute lymphoblastic leukemia. Carcinogenic effects of Aspergillus spp. may not always be mycotoxin related and this requires further investigation.

Carotid distensibility is an indicator of carotid elasticity, which can be used as a cardiovascular disease risk factor. The objective of this study was to investigate how the degree of stenosis influences the strain characteristics of wall and plaque in the internal carotid artery (ICA). Ultrasound videos of the ICA from 83 subjects with atherosclerotic plaques at the origin of the ICA producing variable degrees of stenosis from 10 % to 85 % were investigated. We estimate for each video a carotid diameter during contraction (CDC), a carotid diameter during distension (CDD) in mm and a % of carotid plaque distension (%CPD). We have also estimated the radial strain at wall and plaque (%RSw, %RSp), the longitudinal strain at wall (%LSw) and the shear strain at wall (%SSw) and plaque (%SSp) respectively. The main findings were the following: 1) The %CPD is linearly related to the percentage of ICA stenosis decreasing with an increase in stenosis. 2) There is a statistical significant difference between the % of ICA plaque distensibility of the AS group vs the SY group. 3) There is a statistical significant difference for the RSw, and LSw and %SSw of ICA wall of the AS group vs the SY group. 4) There is a statistical significant difference for the %SSp and %RSp between the AS group vs the SY group. The degree of the ICA stenosis is associated with strain metrics of the carotid wall and plaque. Future work will investigate the proposed methodology on more subjects.

Background COVID‐19 can present with lymphopenia and extraordinary complex multiorgan pathologies that can trigger long‐term sequela. Aims Given that inflammasome products, like caspase‐1, play a role in the pathophysiology of a number of co‐morbid conditions, we investigated caspases across the spectrum of COVID‐19 disease. Materials & Methods We assessed transcriptional states of multiple caspases and using flow cytometry, the expression of active caspase‐1 in blood cells from COVID‐19 patients in acute and convalescent stages of disease. Non‐COVID‐19 subject presenting with various comorbid conditions served as controls. Results Single‐cell RNA‐seq data of immune cells from COVID‐19 patients showed a distinct caspase expression pattern in T cells, neutrophils, dendritic cells, and eosinophils compared with controls. Caspase‐1 was upregulated in CD4+ T‐cells from hospitalized COVID‐19 patients compared with unexposed controls. Post‐COVID‐19 patients with lingering symptoms (long‐haulers) also showed upregulated caspase‐1activity in CD4+ T‐cells that ex vivo was attenuated with a select pan‐caspase inhibitor. We observed elevated caspase‐3/7levels in red blood cells from COVID‐19 patients compared with controls that was reduced following caspase inhibition. Discussion Our preliminary results suggest an exuberant caspase response in COVID‐19 that may facilitate immune‐related pathological processes leading to severe outcomes. Further clinical correlations of caspase expression in different stages of COVID‐19 will be needed. Conclusion Pan‐caspase inhibition could emerge as a therapeutic strategy to ameliorate or prevent severe COVID‐19., This study assesses transcriptional states of multiple caspases and the expression of active caspase‐1 in blood cells from COVID‐19 patients in acute and convalescent stages of disease. Elevated caspase‐3/7 levels in red blood cells is observed in COVID‐19 patients compared to controls. Post‐COVID‐19 patients with lingering symptoms show up‐regulated caspase‐1 activity in CD4+ T‐cells that is attenuated ex vivo with a select pan‐caspase inhibitor. An exuberant caspase response in COVID‐19 that may facilitate immune‐related pathological processes leading to severe outcomes.

A computational workflow which integrates physiologically based kinetic (PBK) modeling, global sensitivity analysis (GSA), approximate Bayesian computation (ABC), and Markov Chain Monte Carlo (MCMC) simulation was developed to facilitate quantitative in vitro to in vivo extrapolation (QIVIVE). The workflow accounts for parameter and model uncertainty within a computationally efficient framework. The workflow was tested using a human PBK model for perfluorooctanoic acid (PFOA) and high throughput screening (HTS) in vitro concentration–response data, determined in a human liver cell line, from the ToxCast/Tox21 database. In vivo benchmark doses (BMDs) for PFOA intake (ng/kg BW/day) and drinking water exposure concentrations (µg/L) were calculated from the in vivo dose responses and compared to intake values derived by the European Food Safety Authority (EFSA). The intake benchmark dose lower confidence limit (BMDL5) of 0.82 was similar to 0.86 ng/kg BW/day for altered serum cholesterol levels derived by EFSA, whereas the intake BMDL5 of 6.88 was six-fold higher than the value of 1.14 ng/kg BW/day for altered antibody titer also derived by the EFSA. Application of a chemical-specific adjustment factor (CSAF) of 1.4, allowing for inter-individual variability in kinetics, based on biological half-life, gave an intake BMDL5 of 0.59 for serum cholesterol and 4.91 (ng/kg BW/day), for decreased antibody titer, which were 0.69 and 4.31 the EFSA-derived values, respectively. The corresponding BMDL5 for drinking water concentrations, for estrogen receptor binding activation associated with breast cancer, pregnane X receptor binding associated with altered serum cholesterol levels, thyroid hormone receptor α binding leading to thyroid disease, and decreased antibody titer (pro-inflammation from cytokines) were 0.883, 0.139, 0.086, and 0.295 ng/ml, respectively, with application of no uncertainty factors. These concentrations are 5.7-, 36-, 58.5-, and 16.9-fold lower than the median measured drinking water level for the general US population which is approximately, 5 ng/ml.

OBJECTIVE To propose a conceptual model to identify points along the condition course where actions or inaction affect downstream burdens of non–cancerous genitourinary conditions (NCGUC). MATERIALS AND METHODS The National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) convened an interdisciplinary meeting to comprehensively consider the burdens of NCGUCs. Subsequently, the authors met monthly to conceptualize the model. RESULTS Inflection points (IP) describe time points during a condition course that are sensitive to change. Our proposed Inflection Point Model (IPM) helps conceptualize burden/benefit trade-offs in any related decision and provides a platform to identify the downstream aggregate burden of a NCGUC across multiple socio-ecological levels at a single time point, which may be summed across the condition course to measure cumulative burden. Two personae demonstrate the utility of this model to better understand impacts of 2 common NCGUCs. CONCLUSION The IPM may be applied in multiple contexts: narrowly to explore burden of a single NCGUC at a single IP; or more broadly, to address multiple conditions, multiple IPs, or multiple domains/levels of social ecology. Applying the IPM may entail combining population data describing prevalence of NCGUCs, associated behaviors, and resulting outcome patterns that can be combined with suitable mathematical models to quantify aggregate and cumulative burden. The IPM challenges stakeholders to expand from the individual to include broader levels of social ecology. Application of the IPM will undoubtedly identify data gaps and research needs that must be fulfilled to delineate and address the burden of NCGUCs.

Objective To provide a conceptual framework to guide investigations into burdens of noncancerous genitourinary conditions (NCGUCs), which are extensive and poorly understood. Methods The National Institute of Diabetes and Digestive and Kidney Diseases convened a workshop of diverse, interdisciplinary researchers and health professionals to identify known and hidden burdens of NCGUCs that must be measured to estimate the comprehensive burden. Following the meeting, a subgroup of attendees (authors of this article) continued to meet to conceptualize burden. Results The Hidden Burden of Noncancerous Genitourinary Conditions Framework includes impacts across multiple levels of well-being and social ecology, including individual (ie, biologic factors, lived experience, behaviors), interpersonal (eg, romantic partners, family members), organizational/institutional (eg, schools, workplaces), community (eg, public restroom infrastructure), societal (eg, health care and insurance systems, national workforce/economic output), and ecosystem (eg, landfill waste) effects. The framework acknowledges that NCGUCs can be a manifestation of underlying biological dysfunction, while also leading to biological impacts (generation and exacerbation of health conditions, treatment side effects). Conclusion NCGUCs confer a large, poorly understood burden to individuals and society. An evidence-base to describe the comprehensive burden is needed. Measurement of NCGUC burdens should incorporate multiple levels of well-being and social ecology, a life course perspective, and potential interactions between NCGUCs and genetics, sex, race, and gender. This approach would elucidate accumulated impacts and potential health inequities in experienced burdens. Uncovering the hidden burden of NCGUCs may draw attention and resources (eg, new research and improved treatments) to this important domain of health.

Nutritional risk screening (NRS) is not yet established in many clinical settings. This study aimed to evaluate the efficacy of two NRS tools, the Paediatric Yorkhill Malnutrition Score (PYMS) and the Screening Tool for the Assessment of Malnutrition in Paediatrics (STAMP), compared to the global dietitians’ clinical judgment. The goal of this study was also to estimate the prevalence of nutritional risk in Greek paediatric patients. Overall, 1506 children, 1–16 years, from paediatric and surgical wards of two Greek hospitals were included. NRS was performed using PYMS and STAMP based either on World Health Organization (WHOGC) or Hellenic growth charts (HGC). The first 907 children were also referred to dietitians who categorized children in low, medium and high nutritional risk according to their global clinical judgment. PYMS, either based on WHOGC or HGC, showed better agreement with dietitians’ feedback (kPYMS_WHO = 0.47, 95%CI: 0.41–0.52, kPYMS_HGC = 0.48, 95%CI: 0.43–0.53) compared to STAMP (kSTAMP_WHO = 0.28, 95%CI: 0.23–0.33, kSTAMP_HGC = 0.26, 95%CI: 0.21–0.32). PYMS also showed the best diagnostic accuracy compared to STAMP in paediatrics and surgical wards separately. Moreover, the PYMS showed similar sensitivity to the STAMP (WHOGC: 82% vs. 84.4%), but a higher positive predictive value (WHOGC: 58.2 vs. 38.7). Using PYMS, high and medium malnutrition risk was observed at 14.9%, and 13.1% of children, respectively. Almost 28% of hospitalised children were at nutritional risk. Children in hospitals should be screened with effective and feasible NRS tools such as PYMS.

COVID-19 can present with lymphopenia and extraordinary complex multi-organ pathologies that can trigger long-term sequela. Given that inflammasome products, like caspase-1, play a role in the pathophysiology of a number of co-morbid conditions, we investigated caspases across the spectrum of COVID-19 disease. We assessed transcriptional states of multiple caspases and using flow cytometry, the expression of active caspase-1 in blood cells from COVID-19 patients in acute and convalescent stages of disease. Non-COVID-19 subjects presenting with various co-morbid conditions served as controls. Single-cell RNA-seq data of immune cells from COVID-19 patients showed a distinct caspase expression pattern in T cells, neutrophils, dendritic cells and eosinophils compared to controls. Caspase-1 was upregulated in CD4+ T-cells from hospitalized COVID-19 patients compared to unexposed controls. Post-COVID-19 patients with lingering symptoms (long-haulers) also showed up-regulated caspase-1 activity in CD4+ T-cells that ex vivo was attenuated with a select pan-caspase inhibitor. We observed elevated caspase-3/7 levels in red blood cells from COVID-19 patients compared to controls that was reduced following caspase inhibition. Our preliminary results suggest an exuberant caspase response in COVID-19 that may facilitate immune-related pathological processes leading to severe outcomes. Further clinical correlations of caspase expression in different stages of COVID-19 will be needed. Pan-caspase inhibition could emerge as a therapeutic strategy to ameliorate or prevent severe COVID-19.

Metabolism is a fundamental cellular mechanism. Its regulation is crucial to maintain cellular homeostasis in response to fluctuation in energy demands and in the availability of nutrients. However, metabolic reactions can be harmful for cells, for example by leading to an increase in oxidative stress or through the generation of toxic by-products, which can in turn damage DNA. To deal with these insults, cells have evolved sophisticated DNA damage response (DDR) pathways that allow for the maintenance of genome integrity. Recent years have seen remarkable progress in unraveling the diverse mechanisms of the DDR. Through such work, it has also emerged that cellular metabolic regulation not only generates DNA damage but also impacts on DNA repair, yet a systematic analysis aimed at identifying interactions between metabolic alterations and the DDR remains unreported. Here, we have functionally explored such interplay, by taking a global and unbiased genetic approach. By utilizing a pooled CRISPR-Cas9 sgRNA library targeting some 3,000 metabolic genes, we have determined the effects on DNA repair following the induction of DNA double-strand breaks induced by the chemotherapeutic etoposide, an inhibitor of topoisomerase 2. Through a phenotypic FACS-based screen with gH2AX used as a marker of DNA damage, we have identified metabolic genes that are required for DNA repair. Candidate genes have been prioritized and validated via an arrayed CRISPR screen based on high-throughput microscopy measuring multiple parameters, across different cell lines and DNA double-strand break inducing agents. We will present data on how metabolic genes impact on DNA repair. Our research sheds light on how the fundamental cellular process of DNA damage maintenance is affected by alterations in cellular metabolism. Moreover, understanding the dynamic interplay between metabolic factors and DDR is also of crucial importance to better design efficient cancer treatments, since cancers are both genetic and metabolic diseases.

Tourette syndrome (TS) is a disabling neurodevelopmental disorder characterized by multiple, recurrent tics. The pharmacological treatment of TS is currently based on dopaminergic antagonists, however, these drugs are associated with extrapyramidal symptoms and other serious adverse events. Recent evidence suggests that positive allosteric modulators (PAMs) of GABAA receptors containing &alpha, 6 subunits (&alpha, 6 GABAARs) oppose the behavioral effects of dopamine. Building on this evidence, in the present study, we tested the efficacy of DK-I-56-1, a highly selective PAM for &alpha, 6 GABAARs, in mouse models of TS exhibiting tic-related responses. DK-I-56-1 significantly reduced tic-like jerks and prepulse inhibition (PPI) deficits in D1CT-7 transgenic mice, a well-documented mouse model of TS. DK-I-56-1 also prevented the exacerbation of spontaneous eyeblink reflex induced by the potent dopamine D1 receptor agonist SKF 82958, a proxy for tic-like responses. We also showed that both systemic and prefrontal cortical administration of DK-I-56-1 countered the PPI disruption caused by SKF 82958. Although the effects of DK-I-56-1 were akin to those elicited by dopaminergic antagonists, this drug did not elicit extrapyramidal effects, as measured by catalepsy. These results point to &alpha, 6 GABAAR PAMs as promising TS therapies with a better safety profile than dopaminergic antagonists.

Background We previously identified that 8/24 COPD patients did not improve dynamic hyperinflation (DH) (DH non-responders) with the application of portable non-invasive ventilation (pNIV; Inspiratory/Expiratory Positive Airway Pressure: 18/8 cmH2O) compared to the pursed lip breathing (PLB) technique during recovery from intermittent exercise (Chynkiamis et al 2020). In the present study we employed Optoelectronic Plethysmography during acute application of pNIV and PLB in recovery from exercise to examine potential differences in the pattern of thoracoabdominal volume regulation between DH responders and DH non-responders. Methods 14 COPD patients (FEV1: 55±21%predicted) performed 2 intermittent cycling trials (consisting of 5 bouts for 2 minutes at 80% of peak work rate interspersed with 2 minutes of recovery) using PLB or pNIV during recovery on a balanced order sequence. Results Patients exhibited two different patterns of response to exercise-induced DH during pNIV compared to PLB application: those who recruited expiratory abdominal muscles, thereby compensating end-expiratory rib cage hyperinflation (DH responders: n=7) and those who did not recruit expiratory abdominal muscles to compensate rib cage hyperinflation (DH non-responders: n=7). In DH responders, pNIV application compared to PLB in the 1st minute of recovery decreased total end-expiratory thoracoabdominal volume by 364±114 ml (p=0.019), secondary to greater reduction in end-expiratory abdominal volume by 338±171 ml (p=0.047). In contrast, in DH non-responders, pNIV application compared to PLB increased end-expiratory thoracoabdominal volume by 379±76 ml (p=0.004), secondary to increased end-expiratory rib cage volume by 348±44 ml (p=0.001) with no change in end-expiratory abdominal volume (31±81 ml; p=0.720). Lung function measures were not different between responders and non-responders. However, DH responders had greater BMI (32.8±6.8) compared to DH non-responders (23.6±4.9) (p=0.019). Conclusions Reports that the respiratory muscles of patients with high BMI might have a mechanical advantage compared to patients with normal BMI (O’Donnell & Ciavaglia, 2014) may partly explain the difference between responders and non-responders. Moreover, pNIV used in the present study provided high extrinsic positive end-expiratory pressure (PEEPe), matching more effectively the higher intrinsic positive end-expiratory pressure, reported in patients with high BMI (O’Donnell & Ciavaglia, 2014). However, PEEPe was likely excessive for the DH non-responders, thereby worsening DH.

Various artificial intelligence (AI) algorithms have been proposed in the literature, that are used as medical assistants in clinical diagnostic tasks. Explainability methods are lighting the black-box nature of these algorithms. The objective of this study was the extraction of rules for the assessment of brain magnetic resonance imaging (MRI) lesions in Multiple Sclerosis (MS) subjects based on texture features. Rule extraction of lesion features was used to explain and provide information on the disease diagnosis and progression. A dataset of 38 subjects diagnosed with a clinically isolated syndrome (CIS) of MS and MRI detectable brain lesions were scanned twice with an interval of 6–12 months. MS lesions were manually segmented by an experienced neurologist. Features were extracted from the segmented MS lesions and were correlated with the expanded disability status scale (EDSS) ten years after the initial diagnosis in order to quantify future disability progression. The subjects were separated into two different groups, G1: EDSS ≤ 3.5 and G2: EDSS > 3.5. Classification models were implemented on the KNIME analytics platform using decision trees (DT), to estimate the models with high accuracy and extract the best rules. The results of this study show the effectiveness of rule extraction as it can differentiate MS subjects with benign course of the disease (G1: EDSS ≤ 3.5) and subjects with advanced accumulating disability (G2: EDSS > 3.5) using texture features. Further work is currently in progress to incorporate argumentation modeling to enable rule combination as well as better explainability. The proposed methodology should also be evaluated on more subjects.

INTRODUCTION: As the second wave of COVID-19 pandemic is in progress the development of fast and cost-effective approaches for diagnosis is essential. The aim of the present study was to develop and evaluate the performance characteristics of a new Bioelectric Recognition Assay (BERA) regarding Sars-CoV-2 detection in clinical samples and its potential to be used as a point of care test. MATERIALS AND METHODS: All tests were performed using a custom portable hardware device developed by EMBIO DIAGNOSTICS (EMBIO DIAGNOSTICS Ltd, Cyprus). 110 positive and 136 negative samples tested by RT-PCR were used in order to define the lower limit of detection (L.O.D.) of the system, as well as the sensitivity and the specificity of the method. RESULTS: The system was able to detect a viral concentration of 4 genome copies/µL. The method displayed total sensitivity of 92.7 % (95 %CI: 86.2-96.8) and 97.8 % specificity (95 %CI: 93.7-99.5). When samples were grouped according to the recorded Ct values the BERA biosensor displayed 100.00 % sensitivity (95 %CI: 84.6-100.0) for Ct values

Common carotid intima–media thickness (CIMT) is a commonly used marker for atherosclerosis and is often computed in carotid ultrasound images. An analysis of different computerized techniques for CIMT measurement and their clinical impacts on the same patient data set is lacking. Here we compared and assessed five computerized CIMT algorithms against three expert analysts’ manual measurements on a data set of 1088 patients from two centers. Inter- and intra-observer variability was assessed, and the computerized CIMT values were compared with those manually obtained. The CIMT measurements were used to assess the correlation with clinical parameters, cardiovascular event prediction through a generalized linear model and the Kaplan–Meier hazard ratio. CIMT measurements obtained with a skilled analyst's segmentation and the computerized segmentation were comparable in statistical analyses, suggesting they can be used interchangeably for CIMT quantification and clinical outcome investigation. To facilitate future studies, the entire data set used is made publicly available for the community at http://dx.doi.org/10.17632/fpv535fss7.1 .

Background The short stability window of several hours from blood collection to measuring basophil activation has limited the use of flow cytometry-based basophil activation assays in clinical settings. We examine if it is possible to extend this window to 1 day allowing for shipment of samples between laboratories. Several options exist for reporting the results including reporting all the measured values directly, calculating ratios and reporting a single value covering all measured results. Each of these options have different stability and value to the physician. Methods Whole blood samples from peanut allergic patients were stimulated with four different peanut concentrations at Day 0, Day 1, and Day 2. Samples were stored under temperature-controlled conditions. Flow cytometry was used to analyze the samples. The basophil activation and degranulation were measured as percentage of positive CD63 basophils and CD203c MFI fold change. Shipped samples were transported under ambient conditions. Results The results show that CD63 is a stable marker at Day 1. The CD203c ratio decreases significantly at Day 1. Calculating the CD63/IgE ratio proves to be more stable than CD63 alone. The most stable readouts are the semi-quantitative results and the trajectory of the dose response curve. Finally, we confirmed that the stability can be extended to samples shipped overnight to the laboratory. Conclusions It is possible to extend the stability of the basophil activation assay to 1 day for samples stored at 18-25°C as well as samples shipped under ambient conditions as long as the temperature is within the 2-37°C range.

Currently, there is no effective vaccine and only one FDA approved early-stage therapy against SARS-CoV-2 infection as an indication to prevent disease progression. Cellular caspases play a role in the pathophysiology of a number of disorders that the co-morbid conditions seen in severe COVID-19 disease. In this study, we assessed transcriptional states of caspases in blood cells from COVID-19 patients. Gene expression levels of select caspases were increased inin vitroSARS-CoV-2 infection models and single cell RNA-Seq data of blood from COVID-19 patients showed a distinct caspase expression in T cells, neutrophils, and dendritic cells. Flow cytometric evaluation of CD4 T cells showed up-regulation of caspase-1 in hospitalized COVID-19 patients compared to unexposed controls. Convalescent COVID-19 patients with lingering symptoms (“long haulers”) showed persistent up-regulation of caspase-1 in CD4 T cells that was attenuatedex vivofollowing co-culture with a select pan-caspase inhibitor. Further, we observed elevated caspase-3 levels in red blood cells from COVID-19 patients compared to controls that were responsive to caspase inhibition. Our results expose an exuberant caspase response in COVID-19 that may facilitate immune-related pathological processes leading to severe outcomes. Pan-caspase inhibition could emerge as a therapeutic strategy to ameliorate or prevent severe COVID-19 outcomes.

Listeria monocytogenes is an intracellular bacterium that causes serious epidemic and sporadic food-borne illnesses in humans. Rapid and trustworthy methods are necessary for the detection of the pathogen to prevent potential food contamination. The aim of this study was to test a newly developed L. monocytogenes biosensor on actual food samples and validate its ability to detect the presence of pathogens robustly and accurately. The newly developed method uses a cell-based biosensor technology (BERA) and a portable device developed by EMBIO Diagnostics called B.EL.D, and provides results within 3 min. Tests were conducted on ready-to-eat lettuce salads, milk and halloumi cheese and the results indicate that the novel system was able to identify inoculated samples with 98%, 90%, and 91% accuracy, respectively. Furthermore, the limit of detection was determined to be as low as 0.6 log CFU mL−1 or g−1 in all food types. Classification of the samples Above or Below the detection limit was accessed through a newly developed algorithm for each food substrate. Samples were also analyzed with the ISO 11290-1:2017 and 11290-2:2017, in parallel. Thus, it was concluded that the newly developed biosensor can be a useful tool in the food supply chain, decreasing the required time for the detection of pathogens and increasing the number of tested samples before they reach the market.

Human food-borne diseases caused by pathogenic bacteria have been significantly increased in the last few decades causing numerous deaths worldwide. The standard analyses used for their detection have significant limitations regarding cost, special facilities and equipment, highly trained staff, and a long procedural time that can be crucial for foodborne pathogens with high hospitalization and mortality rates, such as Listeria monocytogenes. This study aimed to develop a biosensor that could detect L. monocytogenes rapidly and robustly. For this purpose, a cell-based biosensor technology based on the Bioelectric Recognition Assay (BERA) and a portable device developed by EMBIO Diagnostics, called B.EL.D (Bio Electric Diagnostics), were used. Membrane engineering was performed by electroinsertion of Listeria monocytogenes homologous antibodies into the membrane of African green monkey kidney (Vero) cells. The newly developed biosensor was able to detect the pathogen&rsquo, s presence rapidly (3 min) at concentrations as low as 102 CFU mL&minus, 1, demonstrating a higher sensitivity than most existing biosensor-based methods. In addition, lack of cross-reactivity with other Listeria species, as well as with Escherichia coli, was shown, thus, indicating biosensor&rsquo, s significant specificity against L. monocytogenes.

Objective There is a clinical interest in identifying normal appearing white matter (NAWM) areas in brain T2-weighted (T2W) MRI scans in multiple sclerosis (MS) subjects. These areas are susceptible to disease development and areas need to be studied in order to find potential associations between texture feature changes and disease progression. Methods The subjects investigated had a first demyelinating event (Clinically Isolated Syndrome-CIS) at baseline (Time0), and the NAWM0 (i.e. NAWM at Time0) of the brain tissue was subsequently converted to demyelinating plaques (as evaluated in a follow up MRI at Time6–12). 38 untreated subjects that had developed a CIS, had brain MRI scans within an interval of 6–12 months (Time6–12 at follow-up). An experienced MS neurologist manually delineated the demyelinating lesions at Time0 (L0) and at Time6–12 (L6–12). Areas in the Time6–12 MRI scans, where new lesions had been developed, were mapped back to their corresponding NAWM areas on the Time0 MR scans (ROIS0). In addition, contralateral ROIs of similar size and shape were segmented on the same images at Time0 (ROISC0) to form an intra-subject control group. Following that, texture features were extracted from all prescribed areas and MS lesions. Results Texture features were used as input into Support Vector Machine (SVM) models to differentiate between the following: NAWM0 vs ROISC0, NAWM0 vs NAWM6–12, NAWM0 vs L0, NAWM6–12 vs L6–12, ROIS0 vs L0, ROIS0 vs L6–12 and ROIS0 vs ROISC0, where the corresponding % correct classifications scores were 89%, 95%, 98%, 92%, 85%, 90% and 65% respectively. Conclusions Texture features may provide complementary information for following up the development and progression of MS disease. Future work will investigate the proposed method on more subjects.

Tendinopathy accounts for over 30% of primary care consultations and represents a growing healthcare challenge in an active and increasingly ageing population. Recognising critical cells involved in tendinopathy is essential in developing therapeutics to meet this challenge. Tendon cells are heterogenous and sparsely distributed in a dense collagen matrix; limiting previous methods to investigate cell characteristics ex vivo. We applied next generation CITE-sequencing; combining surface proteomics with in-depth, unbiased gene expression analysis of > 6400 single cells ex vivo from 11 chronically tendinopathic and 8 healthy human tendons. Immunohistochemistry validated the single cell findings. For the first time we show that human tendon harbours at least five distinct COL1A1/2 expressing tenocyte populations in addition to endothelial cells, T-cells, and monocytes. These consist of KRT7/SCX+ cells expressing microfibril associated genes, PTX3+ cells co-expressing high levels of pro-inflammatory markers, APOD+ fibro–adipogenic progenitors, TPPP3/PRG4+ chondrogenic cells, and ITGA7+ smooth muscle-mesenchymal cells. Surface proteomic analysis identified markers by which these sub-classes could be isolated and targeted in future. Chronic tendinopathy was associated with increased expression of pro-inflammatory markers PTX3, CXCL1, CXCL6, CXCL8, and PDPN by microfibril associated tenocytes. Diseased endothelium had increased expression of chemokine and alarmin genes including IL33.

A 71-year-old man with end-stage renal disease on hemodialysis presented with bilateral common iliac artery aneurysms diagnosed during the workup of his chronic kidney disease. On computed tomography angiography, common iliac artery aneurysm diameters measured 6.1 cm on the right side and 3.1 cm on the left side. The infrarenal aorta also had a small 3.2-cm aneurysm, but the length from the lowest left renal to the aortic bifurcation was only 6.7 cm, precluding use of most bifurcated endografts. Following an uneventful staged preoperative internal iliac artery embolization, a two-piece D-shaped Altura endograft for the aorta, with bilateral iliac components, landing at the level of the external iliac arteries was successfully performed. Postoperative course was uneventful with no endoleak or endograft migration on computerized tomographic angiography 45 days later, although billowing mimicking an endoleak was evident and will be closely followed.

Targeted cancer therapies represent a milestone towards personalized treatment as they function via inhibition of cancer-specific alterations. Polymerase θ (POLQ), an error-prone translesion polymerase, also involved in DNA double-strand break (DSB) repair, is often upregulated in cancer. POLQ is synthetic lethal with various DNA repair genes, including known cancer drivers such as BRCA1/2, making it essential in homologous recombination-deficient cancers. Thus, POLQ represents a promising target in cancer therapy and efforts for the development of POLQ inhibitors are actively underway with first clinical trials due to start in 2021. This review summarizes the journey of POLQ from a backup DNA repair enzyme to a promising therapeutic target for cancer treatment.

DNA mismatch repair deficiency (dMMR) leads to increased mutation load, which in turn may impact anti-tumor immune responses and treatment effectiveness. Currently, there are different mutational signatures described in primary cancers that are associated with dMMR. Whether the somatic and epigenetic changes in MMR genes precede one or more dMMR signatures, and if so by which mechanism remains unknown. To investigate the relationship between these changes and dMMR signatures, we performed a de novo extraction of mutational signatures in a large cohort of 787 gastric cancer patients. We detected three dMMR-related signatures, one of which clearly discriminates tumors with MLH1 gene silencing caused by hyper-methylation within its promoter (AUC = 98%). We then demonstrate that samples with the highest exposures to signature share features related to better prognosis, encompassing clinical and molecular aspects, as well as altered immune infiltrate composition, predictive of a better response to immune checkpoint inhibitors. Overall, our analysis explored the impact of modifications in MMR-related genes on shaping specific mutational signatures and we provide evidence that patient classification based on mutational signature exposure can identify a group of patients with a good prognosis and who are potentially good candidates for immunotherapy.