Controlled assembly of SNAP-PNA-fluorophore systems on DNA templates to produce fluorescence resonance energy transfer

The SNAP protein is a widely used self-labeling tag that can be used for tracking protein localization and trafficking in living systems. A model system providing controlled alignment of SNAP-tag units can provide a new way to study clustering of fusion proteins. In this work, fluorescent SNAP-PNA conjugates were controllably assembled on DNA frameworks forming dimers, trimers, and tetramers. Modification of peptide nucleic acid (PNA) with the O6-benzyl guanine (BG) group allowed the generation of site-selective covalent links between PNA and the SNAP protein. The modified BG-PNAs were labeled with fluorescent Atto dyes and subsequently chemo-selectively conjugated to SNAP protein. Efficient assembly into dimer and oligomer forms was verified via size exclusion chromatography (SEC), electrophoresis (SDS-PAGE), and fluorescence spectroscopy. DNA directed assembly of homo- and hetero-dimers of SNAP-PNA constructs induced homo- and hetero-FRET, respectively. Longer DNA scaffolds controllably aligned similar fluorescent SNAP-PNA constructs into higher oligomers exhibiting homo-FRET. The combined SEC and homo-FRET studies indicated the 1:1 and saturated assemblies of SNAP-PNA-fluorophore:DNA formed preferentially in this system. This suggested a kinetic/stoichiometric model of assembly rather than binomially distributed products. These BG-PNA-fluorophore building blocks allow facile introduction of fluorophores and/or assembly directing moieties onto any protein containing SNAP. Template directed assembly of PNA modified SNAP proteins may be used to investigate clustering behavior both with and without fluorescent labels which may find use in the study of assembly processes in cells

Ventricular arrhythmias after atrial fibrillation electrical cardioversion: A multicenter study

Background: Ventricular arrhythmias (VAs) after atrial fibrillation (AF) electrical cardioversion (ECV) have been reported. Objective: We sought to assess incidence, timing, and clinical characteristics of patients with post-AF ECV-related VAs. Methods: Multicenter observational retrospective study including 13 centers, incorporating patients with VAs or sudden cardiac death within 10 days of ECV. The total number of ECVs performed during the collecting period was provided. Patients with pre-ECV VAs were excluded. Results: Twenty-three patients with VAs were identified out of 11,897 AF ECVs performed in 13 centers during a median 2-year period, suggesting post-ECV VA incidence of 0.2%. The patients’ mean age was 71 ± 11 years, and 13 (56.5%) were female. AF duration prior to ECV was 71 ± 54 days. Congestive heart failure and hypertension were both found in 17 (74%) patients. QT-prolonging drugs were used by 17 (74%). Index VA occurred 28.5 (interquartile range 5.5–72) hours post-ECV, including torsades de pointes, nonsustained polymorphic ventricular tachycardia, and sudden cardiac death in 17 (74%), 5 (22%), and 1 (4%) patient, respectively. Post-ECV heart rate was slower and QT duration longer compared with pre-ECV (57 ± 11 beats/min vs 113 ± 270 beats/min; P < .001; QT duration 482 ± 61 ms vs 390 ± 60 ms; P < .001). VAs reoccurred in 9 (39%) patients, 11 (interquartile range 3–13.5) hours post–index VA. Two patients had an arrhythmic death within 72 hours post-ECV. Conclusion: VAs post-AF ECV are rare, occur within 3 to 72 hours post-ECV, and are potentially fatal. Our study gives a signal of caution favoring prolonged monitoring in small subset of patients as congestive heart failure patients treated with class III antiarrhythmic drugs, with post-ECV bradycardia, especially (but not exclusively) when QT prolongation noted

The association between statin pre-treatment and LDL Levels on the rate of STEMI presentation: data from the ACS Israeli Survey (ACSIS) 2008–2018

Abstract Background ST-elevation myocardial infarction (STEMI) mostly occurs as a result of vulnerable coronary artery plaque rupture. Constituting hypolipidemic and pleiotropic effects, statins stabilize coronary artery plaque and may prevent STEMI events. Purpose To determine the association between statin pre-treatment and its intensity, low-density-lipoprotein cholesterol (LDL-C) levels and presentation of patients with an ACS (STEMI vs. NSTEMI/unstable angina [UAP]). Methods Data was drawn from the Acute Coronary Syndrome Israeli Survey (ACSIS), a biennial prospective survey of ACS patients hospitalized in all CCU/Cardiology departments during 2008–2018. The rate of STEMI vs NSTEMI/UAP at the time of presentation was calculated by LDL-C levels on admission, stratified to 5 subgroups (&amp;lt;50, 50–69, 70–100, 101–130 and &amp;gt;130 mg/dl) and the use of statins, including statin intensity prior to the index ACS event. Results Among 5,103 patients, 875 (17%) were pre-treated with high intensity statin (HIST), 1,389 (27%) with low intensity statin (LIST), and 2,839 (56%) were statin naive patients. Statin-pre-treated patients were older, more likely to suffer from co-morbidities and cardiovascular diseases and were more often pre-treated with anti-platelets. The proportion of patients presenting with STEMI vs. NSTEMI/UAP was significantly lower among HIST vs. LIST vs. statin naive patients (31.0%, 37.8%, 54.0%, respectively, p for trend &amp;lt;0.001). At each LDL-C level, the proportion of STEMI was significantly lower among HIST vs. LIST vs. statin naive patients. Multivariate analysis adjusting for pertinent variables including propensity score for statin use, revealed that HIST was independently associated with lower STEMI presentation, but LDL-C&amp;lt;70 and LIST were not. Conclusions Among patients admitted with ACS, pre-treatment with high intensity statin but not LDL-C level, was independently associated with a lower probability of presenting with STEMI. Funding Acknowledgement Type of funding source: None </jats:sec

Effect Of Combined Germination, Dehulling And Boiling On Mineral, Sucrose, Stachyose, Fibrulose, And Phytic Acid Content Of Different Chickpea Cultivars

Chickpea is a good source of high quality protein, carbohydrates, vitamins (thiamine and niacin), and minerals. However, its use in industry has been limited by variation in composition with cultivar and also the presence of oligosaccharides, trypsin inhibitors, phytic acids, tannin, and haemagglutinin. Different technologies have been studied to eliminate or minimise the undesirable factors in chickpeas. None of the studied traditional technologies has been found to effectively eliminate or minimise all the undesirable factors in chickpeas. It is not clear whether a combination of these traditional technologies, more especially cooking of germinated and dehulled chickpeas, will significantly reduce all the antinutritional factors. The physical characteristics, stachyose, sucrose, phytic acid, fibrulose, and mineral content of different chickpeas cultivar were determined and compared with reference to infant and child nutrition. The selected cultivars were (1) dehulled and boiled before drying; (2) dehulled followed by soaking and boiling before drying; (3) boiled without dehulling before drying; and germinated, boiled followed by drying and dehulling . The effects of the processing on mineral, sugar, dietary fibre content were evaluated. Desiwere found to have lower seed weight, hydration capacity and swelling capacity compared to kabuli. Seed density, hydration index and swelling index did not vary with cultivar. The mineral density, stachyose, fibrulose, and hull content increased significantly (p<0.05) with the decrease of seed weight whereas phytic acid content did not vary. All processes resulted in an increase in calcium, phosphorous, zinc, and phytic acid and a decrease in potassium, iron, magnesium, sucrose, stachyose and fibrulose content regardless of cultivar type. Germination for 72 hrs followed by boiling, drying and dehulling resulted in highest reduction in antinutritional factors with minimal nutrient loss.It is feasible to use chickpeas as an excellent source of infant follow-on formula/weaning food with minimal mineral fortification and use of low phytic acid cultivars

Effect Of Combined Germination, Dehulling And Boiling On Mineral, Sucrose, Stachyose, Fibrulose, And Phytic Acid Content Of Different Chickpea Cultivars

Chickpea is a good source of high quality protein, carbohydrates, vitamins (thiamine and niacin), and minerals. However, its use in industry has been limited by variation in composition with cultivar and also the presence of oligosaccharides, trypsin inhibitors, phytic acids, tannin, and haemagglutinin. Different technologies have been studied to eliminate or minimise the undesirable factors in chickpeas. None of the studied traditional technologies has been found to effectively eliminate or minimise all the undesirable factors in chickpeas. It is not clear whether a combination of these traditional technologies, more especially cooking of germinated and dehulled chickpeas, will significantly reduce all the antinutritional factors. The physical characteristics, stachyose, sucrose, phytic acid, fibrulose, and mineral content of different chickpeas cultivar were determined and compared with reference to infant and child nutrition. The selected cultivars were (1) dehulled and boiled before drying; (2) dehulled followed by soaking and boiling before drying; (3) boiled without dehulling before drying; and germinated, boiled followed by drying and dehulling . The effects of the processing on mineral, sugar, dietary fibre content were evaluated. Desiwere found to have lower seed weight, hydration capacity and swelling capacity compared to kabuli. Seed density, hydration index and swelling index did not vary with cultivar. The mineral density, stachyose, fibrulose, and hull content increased significantly (p<0.05) with the decrease of seed weight whereas phytic acid content did not vary. All processes resulted in an increase in calcium, phosphorous, zinc, and phytic acid and a decrease in potassium, iron, magnesium, sucrose, stachyose and fibrulose content regardless of cultivar type. Germination for 72 hrs followed by boiling, drying and dehulling resulted in highest reduction in antinutritional factors with minimal nutrient loss.It is feasible to use chickpeas as an excellent source of infant follow-on formula/weaning food with minimal mineral fortification and use of low phytic acid cultivars

Effect of combined germination, dehulling and boiling on mineral, sucrose, stachyose, fibrulose, and phytic acid content of different chickpea cultivars.

Chickpea is a good source of high quality protein, carbohydrates, vitamins (thiamine and niacin), and minerals. However, its use in industry has been limited by variation in composition with cultivar and also the presence of oligosaccharides, trypsin inhibitors, phytic acids, tannin, and haemagglutinin. Different technologies have been studied to eliminate or minimise the undesirable factors in chickpeas. None of the studied traditional technologies has been found to effectively eliminate or minimise all the undesirable factors in chickpeas. It is not clear whether a combination of these traditional technologies, more especially cooking of germinated and dehulled chickpeas, will significantly reduce all the antinutritional factors . The physical characteristics, stachyose, sucrose, phytic acid, fibrulose, and mineral content of different chickpeas cultivar were determined and compared with reference to infant and child nutrition. The selected cultivars were (1) dehulled and boiled before drying; (2) dehulled followed by soaking and boiling before drying; (3) boiled without dehulling before drying; and germinated, boiled followed by drying and dehulling . The effects of the processing on mineral, sugar, dietary fibre content were evaluated. Desi were found to have lower seed weight, hydration capacity and swelling capacity compared to kabuli . Seed density, hydration index and swelling index did not vary with cultivar. The mineral density, stachyose, fibrulose, and hull content increased significantly (p&lt;0.05) with the decrease of seed weight whereas phytic acid content did not vary. All processes resulted in an increase in calcium, phosphorous, zinc, and phytic acid and a decrease in potassium, iron, magnesium, sucrose, stachyose and fibrulose content regardless of cultivar type. Germination for 72 hrs followed by boiling, drying and dehulling resulted in highest reduction in antinutritional factors with minimal nutrient loss. It is feasible to use chickpeas as an excellent source of infant follow -on formula/weaning food with minimal mineral fortification and use of low phytic acid cultivars.</jats:p

D-dimer as a prognostic factor in patients admitted to a tertiary care intensive coronary care unit

Abstract Introduction D-dimer is a small protein fragment and is a product of fibrinolysis. A high levels of D-dimer have been suggested as a prognostic factor in cancerous and other critically ill patients. We aimed to evaluate D-dimer levels and outcomes of critically ill patients admitted to a tertiary care intensive coronary care unit (ICCU). Material and method All patients admitted to the ICCU at our Medical Center between January 1, 2020 and December 31, 2020 were included in the study. Patients were divided into 2 groups according to their D-dimer level on admission. Low D-dimer level &amp;lt;500 ng/ml, and high D-dimer level ≥500 ng/ml. Survival, in-hospital interventions and complications were compared. Results and discussion Overall 1,082 consecutive patients were included, mean age was 67 (±16), 70% were males. Of them 296 (27.4%) had low D-dimer level and 663 (61.3%) had high D-dimer level. Patients with high D-dimer level were older as compared to patients with low D-dimer level (mean age 70.4±15 and 59±13 years respectively, p=0.004), had significantly higher rate of female gender (35.9% vs 15.9% respectively, p&amp;lt;0.0001) and significantly higher rate of any prior cardiac interventions prior to their admission (26.7% vs 4.4% respectively, p&amp;lt;0.0001). Interestingly, patients with high D-dimer level had significantly lower rate of any acute coronary syndrome (ACS) as compared with the low D-dimer group (25.7 vs 66.4% respectively, p&amp;lt;0.0001) and lower rate of smokers (22.5 vs 45.6% respectively, p&amp;lt;0.0001). All 11 post-COVID-19 patients had high D-dimer level on admission. A multivariate Cox proportional hazards analysis for mortality, adjusted for age, gender, risk factors for cardiovascular disease, ejection fraction&amp;lt;40 found that high D-dimer level was independently associated with higher mortality rates (HR=5.8; 95% CI; 1.7–19.1; p=0.004) as shown in Figure 1. Conclusion Elevated D-dimer levels on admission in ICCU patients is a poor prognostic factor of in-hospital morbidity and mortality in the first year following hospitalization. Funding Acknowledgement Type of funding sources: None. Cumulative survival according to d-Dimer </jats:sec

Association between cardiac pathology and outcomes of patients with COVID-19 using a hand-held ultrasound

Abstract Background The association between COVID-19 infection and the cardiovascular system has been well described. Strict precautions limit the use of formal echocardiography in this setting. Information on the importance of the utilization of a hand-held point-of-care cardiac ultrasound (POCCUS) for cardiac evaluation in these patients is scarce. Objective To investigate the utilization of hand-held echocardiography in COVID-19 hospitalized patients and the association between cardiac pathologies and outcomes. Methods Consecutive patients diagnosed with COVID-19 underwent POCCUS evaluation using a hand-held ultrasound within 24 hours of admission at our institute, throughout March-May 2020. According to the POCCUS results, the patients were divided into two groups: 'Normal' and 'Abnormal' (including left or right ventricular dysfunction or enlargement, or moderate/severe valvular regurgitation/stenosis). Results Among 102 patients, 26 (25.5%) had an abnormal POCCUS study. They were older, with more co-morbidities, cardiovascular disease history, chronic medical therapy, and more severe presenting symptoms, as compared to the group with a normal echocardiography exam. Individual and composite endpoints (advanced ventilatory support, acute decompensated heart failure, shock, or death) are presented in Table 1. Multivariate logistic regression analysis adjusting for pertinent variables revealed that abnormal echocardiography at presentation was independently associated with the composite endpoint OR=4.63 (95% CI 1.51–14.15, p=0.007). Conclusions Abnormal echocardiography results in COVID-19 infection settings are associated with a higher burden of medical comorbidities and independently predict major adverse endpoints. Hand-held POCCUS at presentation can be utilized as an important tool for risk stratification for hospitalized COVID-19 patients. Funding Acknowledgement Type of funding sources: Private hospital(s). Main funding source(s): SZMC Scientific </jats:sec