The occurrence of immune-evasive SARS-CoV-2 strains emphasizes the importance to search for broad-acting antiviral compounds. Our previous
study showed that
root extract EPs
7630 has combined ...antiviral and immunomodulatory properties in SARS-CoV-2-infected human lung cells. Here we assessed
effects of EPs
7630 in SARS-CoV-2-infected hamsters, and investigated properties of EPs
7630 and its functionally relevant constituents in context of phenotypically distinct SARS-CoV-2 variants. We show that EPs
7630 reduced viral load early in the course of infection and displayed significant immunomodulatory properties positively modulating disease progression in hamsters. In addition, we find that EPs
7630 differentially inhibits SARS-CoV-2 variants in nasal and bronchial human airway epithelial cells. Antiviral effects were more pronounced against Omicron BA.2 compared to B.1 and Delta, the latter two preferring TMPRSS2-mediated fusion with the plasma membrane for cell entry instead of receptor-mediated low pH-dependent endocytosis. By using SARS-CoV-2 Spike VSV-based pseudo particles (VSVpp), we confirm higher EPs
7630 activity against Omicron Spike-VSVpp, which seems independent of the serine protease TMPRSS2, suggesting that EPs
7630 targets endosomal entry. We identify at least two molecular constituents of EPs
7630, i.e., (-)-epigallocatechin and taxifolin with antiviral effects on SARS-CoV-2 replication and cell entry. In summary, our study shows that EPs
7630 ameliorates disease outcome in SARS-CoV-2-infected hamsters and has enhanced activity against Omicron, apparently by limiting late endosomal SARS-CoV-2 entry.
Summary
Based on case series, potency rates after radical prostatectomy (RPE) differ substantially and – furthermore – it remains unclear whether they have improved in more recent surgical series. ...The purpose of this study was to investigate whether potency rates after RPE have improved over the years. A systematic analysis of the control arms of all randomized controlled trials (RCT; n = 11) on penile rehabilitation after RPE was carried out. In total, 2009 patients were included in these RCTs, 685 thereof in the respective control arms, who were either observed or received placebo. Assessment of erectile function in these studies was carried out by the Sexual Encounter Profile (SEP) or the International Index of Erectile Function (IIEF). Eight trials used SEP3 as study endpoint. The rate of positive response to SEP3 (=erectile function sufficient for successful intercourse) in the control arms was 20% in 1997 (year of publication), 10% in 2003, 19% in 2004, 25% in 2008, 21% in 2010, 67% in 2011, 10% in 2013, and 22% in 2014. Eight RCTs assessed the IIEF‐EF, yet results were not reported uniformly. In the control arms the IIEF‐EF was 9.2 (year of publication 2003), 13.3 (2004), 8.8 (2008), 25% ≥22.0 (2008), 17.4 (2010), 58% ≥26.0 (2011), 9.3 (2013), and 11.6 (2014). Limitations of this analysis are a positive selection bias regarding patient recruitment, surgical approach, and the non‐uniform inclusion and outcome criteria. This systematic analysis of the control arms of all RCTs on penile rehabilitation after nerve‐sparing RPE shows (i) that the rate of undisturbed erectile function is in the range 20–25% in most studies and (ii) that these rates have not substantially improved or changed over the past 17 years.
BACKGROUND: Healthy cows consume large amounts of potassium and a sudden loss in appetite can lead to hypokalemia. The routine method to evaluate potassium homeostasis is the measurement of the ...extracellular potassium in plasma or serum, but this does not provide information about the intracellular potassium pool. HYPOTHESIS/OBJECTIVES: To evaluate potassium homeostasis by comparing the extracellular and intracellular potassium concentration in cows with reduced feed intake and gastrointestinal ileus. ANIMALS: Twenty cows 1–3 days postpartum (group 1) and 20 cows with gastrointestinal ileus (group 2). METHODS: Observational cross‐sectional study. Plasma potassium was measured by using an ion‐sensitive electrode. Intracellular potassium was measured in erythrocytes and muscle tissue (muscle biopsy) by using inductively coupled plasma optical emission spectroscopy. RESULTS: Cows of group 1 did not have hypokalemia. Overall cows with gastrointestinal ileus were hypokalemic (mean ± SD, 2.9 mmol/L ± 0.78), but potassium concentration in erythrocytes and muscle tissue was not lower than in postpartum cows. Intracellular potassium in erythrocytes varied very widely; group 1: 3497–10735 mg/kg (5559 ± 2002 mg/kg), group 2: 4139–21678 mg/kg (7473 ± 4034 mg/kg). Potassium in muscle tissue did not differ between group 1 (3356 ± 735 mg/kg wet weight) and group 2 (3407 ± 1069 mg/kg wet weight). No association between extracellular and intracellular potassium concentrations was detected. CONCLUSIONS AND CLINICAL IMPORTANCE: That measurement of plasma potassium concentration is not sufficient to evaluate potassium metabolism of cows.
Sirtuins (SIRT) are ubiquitous histone and protein deacetylases and a member of this family, SIRT1, is the best-studied one. Its functions in endothelial cells encompass branching angiogenesis, ...activation of endothelial nitric oxide synthase, regulation of proapoptotic and proinflammatory pathways, among others. Defective SIRT1 activity has been described in various cardiovascular, renal diseases and in aging-associated conditions. Therefore, understanding of SIRT1-deficient, endothelial dysfunctional phenotype has much to offer clinically. Here, we summarize recent studies by several investigative teams of the characteristics of models of global endothelial SIRT1 deficiency, the causes of facilitative development of fibrosis in these conditions, dissect the protein composition of the aberrant secretome of SIRT1-deficient endothelial cells and present several components of this aberrant secretome that are involved in fibrogenesis via activation of fibroblasts to myofibroblasts. These include ligands of Wnt and Notch pathways, as well as proteolytic fragments of glycocalyx core protein, syndecan-4. The latter finding is crucial for understanding the degradation of glycocalyx that accompanies SIRT1 deficiency. This spectrum of abnormalities associated with SIRT1 deficiency in endothelial cells is essential for understanding the origins and features of endothelial dysfunction in a host of cardiovascular and renal diseases.
Divertor target is one of the most critical in-vessel components in a fusion power plant being in charge of particle and power exhaust. The targets are exposed to severe thermal loads produced by ...steady bombardment of impinging plasma flux. Since 2014, integrated R&D efforts have been continued aiming at developing a design concept and high-heat-flux (HHF) technologies for divertor targets of the European DEMO reactor. Recently, the second round (2017–2019) of the R&D program has been concluded. As in the first R&D round, five water-cooled target design concepts were further developed and evaluated. Fabrication technologies were improved reaching a consolidated production quality. Extensive HHF tests were conducted using small-scale mock-ups for extended loading regimes (heat flux: 20–32MW/m²). Comparative studies were performed to investigate effects of copper interlayer thickness (0.1–1 mm) and different tungsten armor materials. In the present paper, the final results of the second round HHF testing campaign are reported. The HHF performance of each design variant is discussed based on in-situ diagnostic data (infrared thermography), ultrasonic inspection images and postmortem metallographic micrographs. All monoblock-type design concepts passed the specified qualification criterion (≥500 pulses at 20 MW/m², coolant: 130 °C) without any failure or armor cracking. Moreover, two of them (ITER-like and composite pipe) remained fully intact even under 25 MW/m² (100 pulses) and 32 MW/m² (5 pulses).
Rheumatoid arthritis (RA) is a common autoimmune disease typically affecting joints symmetrically. A small number of patients develop unilateral and severely destructive wrist arthritis (DWA). The ...objective of our study was to characterise patients with this type of affection.
This was a retrospective cohort study of RA patients with positive RF/anti-CCP antibodies. Clinical characteristics, including, age, gender, disease duration, dexterity, occupational history, smoking status, and the number of prescribed DMARDs were recorded. Conventional radiographs were evaluated using the modified Sharp/van der Heijde scoring (mSS) method.
We analysed our laboratory database of 1247 patients and identified 559 patients with a clinical diagnosis of RA. For 395 of the patients, radiographs of the hands were available for evaluation. 25 patients had extensive unilateral DWA, corresponding to a prevalence of 6.3% (25 of 395 patients). 11 patients were excluded due to incomplete data. Of the remaining 14 patients, 13 were female with a median age of 61 (33-83) years, and median disease duration of 18 (1-33) years. 8 of 11 (72.7%) patients were smokers; in three, smoking status was not known. 80% with known dexterity developed unilateral DWA in the dominant hand. Total mSS was significantly higher on the affected side (39, interquartile range 35.25-46.25) versus non-affected (13, IQR 3-23). MSS were not different if the carpal bones were excluded from scoring. Side of involvement (left vs. right), or dominant versus non-dominant hand, did not result in a different mSS.
Unilateral DWA is a rare variant of RA which predominantly affects women who smoke.
Amyloid fibrils are filamentous protein aggregates implicated in several common diseases such as Alzheimer’s disease and type II diabetes. Similar structures are also the molecular principle of the ...infectious spongiform encephalopathies such as Creutzfeldt–Jakob disease in humans, scrapie in sheep, and of the so-called yeast prions, inherited non-chromosomal elements found in yeast and fungi. Scanning transmission electron microscopy (STEM) is often used to delineate the assembly mechanism and structural properties of amyloid aggregates. In this review we consider specifically contributions and limitations of STEM for the investigation of amyloid assembly pathways, fibril polymorphisms and structural models of amyloid fibrils. This type of microscopy provides the only method to directly measure the mass-per-length (MPL) of individual filaments. Made on both
in vitro assembled and
ex vivo samples, STEM mass measurements have illuminated the hierarchical relationships between amyloid fibrils and revealed that polymorphic fibrils and various globular oligomers can assemble simultaneously from a single polypeptide. The MPLs also impose strong constraints on possible packing schemes, assisting in molecular model building when combined with high-resolution methods like solid-state nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR).
The enzyme pyruvate kinase M2 (PKM2) plays a major role in the switch of tumor cells from oxidative phosphorylation to aerobic glycolysis, one of the hallmarks of cancer. Different allosteric ...inhibitors or activators and several posttranslational modifications regulate its activity. Head and neck squamous cell carcinoma (HNSCC) is a common disease with a high rate of recurrence. To find out more about PKM2 and its modulation in HNSCC, we examined a panel of HNSCC cells using real-time cell metabolic analysis and Western blotting with an emphasis on phosphorylation variant Tyr105 and two reagents known to impair PKM2 activity. Our results show that in HNSCC, PKM2 is commonly phosphorylated at Tyrosine 105. Its levels depended on tyrosine kinase activity, emphasizing the importance of growth factors such as EGF (epidermal growth factor) on HNSCC metabolism. Furthermore, its correlation with the expression of CD44 indicates a role in cancer stemness. Cells generally reacted with higher glycolysis to PKM2 activator DASA-58 and lower glycolysis to PKM2 inhibitor Compound 3k, but some were more susceptible to activation and others to inhibition. Our findings emphasize the need to further investigate the role of PKM2 in HNSCC, as it could aid understanding and treatment of the disease.
Photoabsorption by and fluorescence of the Kα transitions in highly charged iron ions are essential mechanisms for x-ray radiation transfer in astrophysical environments. We study photoabsorption due ...to the main Kα transitions in highly charged iron ions from heliumlike to fluorinelike (Fe24+ to Fe17+) using monochromatic x rays around 6.6 keV at the PETRA III synchrotron photon source. Natural linewidths were determined with hitherto unattained accuracy. The observed transitions are of particular interest for the understanding of photoexcited plasmas found in x-ray binary stars and active galactic nuclei.
Summary
By quantitative immunoblot analyses and scanning transmission electron microscopy (STEM), we determined that the needle of the Yersinia enterocolitica E40 injectisome consists of 139 ± 19 ...YscF subunits and that the tip complex is formed by three to five LcrV monomers. A pentamer represented the best fit for an atomic model of this complex. The N‐terminal globular domain of LcrV forms the base of the tip complex, while the central globular domain forms the head. Hybrids between LcrV and its orthologues PcrV (Pseudomonas aeruginosa) or AcrV (Aeromonas salmonicida) were engineered and recombinant Y. enterocolitica expressing the different hybrids were tested for their capacity to form the translocation pore by a haemolysis assay. There was a good correlation between haemolysis, insertion of YopB into erythrocyte membranes and interaction between YopB and the N‐terminal globular domain of the tip complex subunit. Hence, the base of the tip complex appears to be critical for the functional insertion of YopB into the host cell membrane.
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