Single-molecule techniques have been developed for commercial DNA sequencing1,2. One emerging strategy uses a nanopore to analyze DNA molecules as they are driven electrophoretically in single file order past a sensor3-5. However, uncontrolled DNA strand electrophoresis through nanopores is too fast for accurate base reads6. A proposed solution would employ processive enzymes to deliver DNA through the pore at a slower average rate7. Here, we describe forward and reverse ratcheting of DNA templates through the α–hemolysin (α-HL) nanopore controlled by wild-type phi29 DNA polymerase (phi29 DNAP). DNA strands were examined in single file order at one nucleotide spatial precision in real time. The registry error probability (either an insertion or deletion during one pass along a template strand) ranged from 10% to 24.5% absent optimization. This general strategy facilitates multiple reads of individual template strands and is transferrable to other nanopore devices for implementation of DNA sequence analysis.
epicardial adipose tissue (eAt) is associated with cardiovascular risk. the longitudinal change in eAt volume (eAtv) and density (eAtd), and potential modulators of these parameters, has not been described. We prospectively recruited 90 patients with non-obstructive coronary atherosclerosis on baseline computed tomography coronary angiography (ctcA) performed for suspected coronary artery disease to undergo a repeat research CTCA. EATv in millilitres (mL) and EATd in Hounsfield units (HU) were analysed and multivariable regression analysis controlling for traditional cardiovascular risk factors (cVRf) performed to assess for any predictors of change. Secondary analysis was performed based on statin therapy. The median duration between CTCA was 4.3years. Mean EATv increased at follow-up (72 ± 33 mL to 89 ± 43 mL, p < 0.001) and mean EATd decreased (baseline −76 ± 6 HU vs. −86 ± 5 HU, p < 0.001). There were no associations between baseline variables of body mass index, age, sex, hypertension, hyperlipidaemia, diabetes or smoking on change in EATv or EATd. No difference in baseline, follow-up or delta EATv or EATd was seen in patients with (60%) or without baseline statin therapy. in this select group of patients, eAtv consistently increased and eAtd consistently decreased at long-term follow-up and these changes were independent of cVRf, age and statin use. together with the knowledge of strong associations between EAT and cardiac disease, these findings may suggest that eAt is an independent parameter rather than a surrogate for cardiovascular risk. open Scientific RepoRtS | (2020) 10:7109 | https://doi.org/10.1038/s41598-020-63135-z www.nature.com/scientificreports www.nature.com/scientificreports/ Scientific RepoRtS | (2020) 10:7109 | https://doi.org/10.1038/s41598-020-63135-zwww.nature.com/scientificreports www.nature.com/scientificreports/ however, this is reflective of the current literature in examining relevant associations of EAT. Finally, there is potential for error in using delta EAT values with potential overlap from test-retest variability. Our previous work has demonstrated limits of agreement up to 10 mL higher or lower between observers with a mean bias however of only 1 mL, however our inter-observer correlation was excellent at 0.98 with assessors blinded to scan timing and patient details. conclusion Epicardial adipose tissue volume and density demonstrate significant longitudinal changes in patients with non-obstructive coronary artery disease with a consistent increase in EAT volume and consistent decrease in EAT density. There are no clinical risk factors that appear to associate with the change in EAT parameters and this effect is also independent of statin therapy. This finding may suggest that EAT is an independent marker, rather than surrogate of cardiovascular risk.
Sepsis is a substantial healthcare burden, and its management continues to be a major challenge. Prior studies demonstrate conflicting evidence regarding the utility of vitamin C in sepsis. This systematic review and meta-analysis aim to collect data among critically ill patients (sepsis/septic shock), comparing the efficacy of parenteral vitamin C with standard care.A literature review was conducted using databases including PubMed, Web of Science, Google Scholar, and the Cochrane Library to identify randomized controlled trials (RCTs) and observational studies comparing intravenous vitamin C alone or in combination with thiamine or glucocorticoids to the standard of care. We identified 11 RCTs and seven retrospective cohort studies. The primary outcome was 28-day mortality. Secondary outcomes included intensive care unit (ICU) length of stay, change in Sequential Organ Failure Assessment (SOFA) score, duration of vasopressor use, and duration of mechanical ventilation.A total of 18 studies with 4078 patients were included in our final analysis. Overall, we found no mortality reduction in patients treated with vitamin C compared to standard of care (odds ratio (OR) 0.92; 95% confidence interval (CI) 0.78 to 1.09; p = 0.34). Studies that reported a change in SOFA scores, ICU length of stay, duration of mechanical ventilation, or duration of vasopressor use did not show any significant difference between groups. Subgroup analysis with RCT versus observational studies and vitamin C dosage regimens did not show any difference.Among patients with sepsis or septic shock, treatment with vitamin C was not associated with a reduction in mortality, ICU length of stay, change in SOFA score, duration of vasopressor use, or duration of mechanical ventilation. Further studies are needed to demonstrate the potential role of vitamin C in the management of sepsis.
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