Quantitative viral load assays have transformed our understanding of viral diseases. They hold similar potential to advance COVID-19 control and prevention, but SARS-CoV-2 viral load tests are not yet widely available. SARS-CoV-2 molecular diagnostic tests, which typically employ real-time RT-PCR, yield semiquantitative results only. Droplet digital RT-PCR (RT-ddPCR) offers an attractive platform for SARS-CoV-2 RNA quantification. Eight primer/probe sets originally developed for real-time RT-PCR-based SARS-CoV-2 diagnostic tests were evaluated for use in RT-ddPCR; three were identified as the most efficient, precise, and sensitive for RT-ddPCR-based SARS-CoV-2 RNA quantification. For example, the analytical efficiency for the E-Sarbeco primer/probe set was approximately 83%, whereas assay precision, measured as the coefficient of variation, was approximately 2% at 1000 input copies/reaction. Lower limits of quantification and detection for this primer/probe set were 18.6 and 4.4 input SARS-CoV-2 RNA copies/reaction, respectively. SARS-CoV-2 RNA viral loads in a convenience panel of 48 COVID-19-positive diagnostic specimens spanned a 6.2log10 range, confirming substantial viral load variation in vivo. RT-ddPCR-derived SARS-CoV-2 E gene copy numbers were further calibrated against cycle threshold values from a commercial real-time RT-PCR diagnostic platform. This log-linear relationship can be used to mathematically derive SARS-CoV-2 RNA copy numbers from cycle threshold values, allowing the wealth of available diagnostic test data to be harnessed to address foundational questions in SARS-CoV-2 biology.This study leveraged a gene-protein assay to assess MYC and PTEN status at prostate cancer biopsy and examined the association with adverse outcomes after surgery. MYC gain and PTEN loss were simultaneously assessed by chromogenic in situ hybridization and immunohistochemistry, respectively, using 277 Grade Group 2 needle biopsies that were followed by prostatectomy. The maximal size of cribriform Gleason pattern 4 carcinoma (CRIB), the presence of intraductal carcinoma (IDC), and percentage of Gleason pattern 4 carcinoma at biopsy were also annotated. MYC gain or PTEN loss was present in 19% and 18% of biopsies, respectively, whereas both alterations were present in 9% of biopsies. Tumors with one or both alterations were significantly more likely to have non-organ-confined disease (NOCD) at radical prostatectomy. In logistic regression models, including clinical stage, tumor volume on biopsy, and presence of CRIB/IDC, cases with MYC gain and PTEN loss remained at higher risk for NOCD (odds ratio, 6.23; 95% CI, 1.74-24.55; P = 0.005). The area under the curve for a baseline model using CAPRA variables (age, prostate-specific antigen, percentage of core involvement, clinical stage) was increased from 0.68 to 0.69 with inclusion of CRIB/IDC status and to 0.75 with MYC/PTEN status. Dual MYC/PTEN status can be assessed in a single slide and is independently associated with increased risk of NOCD for Grade Group 2 biopsies.
  Eccentric (ECC) cycling is viewed as an alternative to concentric (CON) cycling for exercise training in patients with severe COPD as it induces a much lower ventilatory demand for a given mechanical load than CON cycling. However, a more hyperpneic breathing pattern (i.e., higher f
  and lower tidal volume (VT)) during ECC than during CON has been reported in healthy subjects.
 
  Do patients with severe COPD develop a more hyperpneic breathing pattern during ECC than during CON cycling, and is it associated with differences in dynamic hyperinflation, ventilatory efficiency and cardiometabolic adaptation?
 
  Fourteen patients with severe COPD performed incremental CON and ECC cardiopulmonary exercise tests (CPET). Several physiological parameters including VT, f
  , inspiratory capacity (IC) and oxygen consumption (V̇O
  ) were recorded at each workload increment during CPET.
 
  At the highest identical minute ventilation (V̇E) achieved during ECC and CON (28.6 ± 4.6 L.min
  ), VT was lower (1010 ± 218 vs. 1100 ± 233 mL; p = 0.02), f
  was higher (29.0 ± 5.1 vs. 27.0 ± 5.5 min
  ; p = 0.03), IC(% baseline) was lower (84 ± 10 vs. 78 ± 9; p < 0.01) and markers of ventilatory efficiency were poorer during ECC than during CON. Similar results were found at the highest identical V̇O
  achieved during ECC and CON.
 
  The finding of a more hyperpneic ventilatory pattern during ECC cycling together with a lower IC and a poorer ventilatory efficiency suggests that ECC exercise training should be decided with caution in patients with severe COPD.
 The finding of a more hyperpneic ventilatory pattern during ECC cycling together with a lower IC and a poorer ventilatory efficiency suggests that ECC exercise training should be decided with caution in patients with severe COPD.Balb/c mice respiratory mechanics was studied in two intravenous methacholine (MCh) protocols bolus and continuous infusion. The Constant Phase Model (CPM) was used in this study. The harmonic distortion index (kd) was used to assess the respiratory system nonlinearity. The analysis of variance showed difference between groups (OVA vs control) and among doses for both protocols. Bolus protocol posttest there was a difference between OVA and control at 0.3 and 1 mg/kg doses (p less then 0.0001 and p less then 0.001) for Rn. Infusion there was a difference between OVA and control at 192 μg.kg-1.min-1 dose for Rn, G and H, (p less then 0.01; p less then 0.001; p less then 0.001). An increment was found in kd values near to the observed peak values in bolus protocol.  https://www.selleckchem.com/products/ipi-145-ink1197.html  The bolus protocol could better differentiate inflamed and non-inflamed airway resistance, whereas the differences between OVA and control in continuous infusion protocol were associated to airway- and, mainly, parenchyma-related parameters. Moreover, the bolus protocol presented a higher nonlinear degree compared to the infusion protocol.
  How benign liver steatosis progresses to nonalcoholic steatohepatitis (NASH), fibrosis, and hepatocellular carcinoma (HCC) remains elusive. NASH progression entails diverse pathogenic mechanisms and relies on complex cross-talk between multiple tissues such as the gut, adipose tissues, liver, and the brain. Using a hyperphagic mouse fed with a Western diet (WD), we aimed to elucidate the cross-talk and kinetics of hepatic and extrahepatic alterations during NASH-HCC progression, as well as regression.
 
  Hyperphagic mice lacking a functional Alms1 gene (Foz/Foz) and wild-type littermates were fed WD or standard chow for 12 weeks for NASH/fibrosis and for 24 weeks for HCC development. NASH regression was modeled by switching back to normal chow after NASH development.
 
  Foz+WD mice were steatotic within 1 to 2 weeks, developed NASH by 4 weeks, and grade 3 fibrosis by 12 weeks, accompanied by chronic kidney injury. Foz+WD mice that continued on WD progressed to cirrhosis and HCC within 24 weeks and had reduced survival as a result of cardiac dysfunction.