MUC16 Is Overexpressed in Idiopathic Pulmonary Fibrosis and Induces Fibrotic Responses Mediated by Transforming Growth Factor-β1 Canonical Pathway

Several transmembrane mucins have demonstrated that they contribute intracellularly to induce fibrotic processes. The extracellular domain of MUC16 is considered as a biomarker for disease progression and death in IPF patients. However, there is no evidence regarding the signalling capabilities of MUC16 that contribute to IPF development. Here, we demonstrate that MUC16 was overexpressed in the lung tissue of IPF patients (n = 20) compared with healthy subjects (n = 17) and localised in fibroblasts and hyperplastic alveolar type II cells. Repression of MUC16 expression by siRNA-MUC16 transfection inhibited the TGF-β1-induced fibrotic processes such as mesenchymal/ myofibroblast transformations of alveolar type II A549 cells and lung fibroblasts, as well as fibroblast proliferation. SiRNA-MUC16 transfection also decreased the TGF-β1-induced SMAD3 phosphorylation, thus inhibiting the Smad Binding Element activation. Immunoprecipitation assays and confocal immunofluorescence showed the formation of a protein complex between MUC16/p-SMAD3 in the cell membrane after TGF-β1 stimulation. This study shows that MUC16 is overexpressed in IPF and collaborates with the TGF-β1 canonical pathway to induce fibrotic processes. Therefore, direct or indirect targeting of MUC16 could be a potential drug target for human IPF.     

Metal-Bound Methisazone; Novel Drugs Targeting Prophylaxis and Treatment of SARS-CoV-2, a Molecular Docking Study

SARS-CoV-2 currently lacks effective first-line drug treatment. We present promising data from in silico docking studies of new Methisazone compounds (modified with calcium, Ca; iron, Fe; magnesium, Mg; manganese, Mn; or zinc, Zn) designed to bind more strongly to key proteins involved in replication of SARS-CoV-2. In this in silico molecular docking study, we investigated the inhibiting role of Methisazone and the modified drugs against SARS-CoV-2 proteins: ribonucleic acid (RNA)-dependent RNA polymerase (RdRp), spike protein, papain-like protease (PlPr), and main protease (MPro). We found that the highest binding interactions were found with the spike protein (6VYB), with the highest overall binding being observed with Mn-bound Methisazone at −8.3 kcal/mol, followed by Zn and Ca at −8.0 kcal/mol, and Fe and Mg at −7.9 kcal/mol. We also found that the metal-modified Methisazone had higher affinity for PlPr and MPro. In addition, we identified multiple binding pockets that could be singly or multiply occupied on all proteins tested. The best binding energy was with Mn–Methisazone versus spike protein, and the largest cumulative increases in binding energies were found with PlPr. We suggest that further studies are warranted to identify whether these compounds may be effective for treatment and/or prophylaxis.     

Effect of microbial transglutaminase on the functional properties of megrim (Lepidorhombus boscii) skin gelatin

This paper examines the effect of a microbial transglutaminase (TGase) on the gelling and viscoelastic properties of a gelatin from megrim (Lepidorhombus boscii) skins. Although TGase extended the setting time of fish gelatin, it was found that melting temperature, gel strength and viscosity in solution at 60 °C were considerably increased by the covalent cross‐linking action of the enzyme, as observed by SDS‐PAGE and SEM. Increasing concentrations of TGase increased the elasticity and cohesiveness of gelatin gels but reduced gel strength and hardness. Partial inactivation of the enzyme was achieved thermally without negatively affecting the properties of the gelatin; whether or not gelatin is thermoreversible depends essentially on the degree of enzyme inactivation. © 2001 Society of Chemical Industry     

PEDOT-Carbon Nanotube Counter Electrodes and Bipyridine Cobalt (II/III) Mediators as Universally Compatible Components in Bio-Sensitized Solar Cells Using Photosystem I and Bacteriorhodopsin

In nature, solar energy is captured by different types of light harvesting protein–pigment complexes. Two of these photoactivatable proteins are bacteriorhodopsin (bR), which utilizes a retinal moiety to function as a proton pump, and photosystem I (PSI), which uses a chlorophyll antenna to catalyze unidirectional electron transfer. Both PSI and bR are well characterized biochemically and have been integrated into solar photovoltaic (PV) devices built from sustainable materials. Both PSI and bR are some of the best performing photosensitizers in the bio-sensitized PV field, yet relatively little attention has been devoted to the development of more sustainable, biocompatible alternative counter electrodes and electrolytes for bio-sensitized solar cells. Careful selection of the electrolyte and counter electrode components is critical to designing bio-sensitized solar cells with more sustainable materials and improved device performance. This work explores the use of poly (3,4-ethylenedioxythiophene) (PEDOT) modified with multi-walled carbon nanotubes (PEDOT/CNT) as counter electrodes and aqueous-soluble bipyridine cobalt^II/III complexes as direct redox mediators for both PSI and bR devices. We report a unique counter electrode and redox mediator system that can perform remarkably well for both bio-photosensitizers that have independently evolved over millions of years. The compatibility of disparate proteins with common mediators and counter electrodes may further the improvement of bio-sensitized PV design in a way that is more universally biocompatible for device outputs and longevity.     

N-acetylcysteine Reduces Inflammasome Activation Induced by SARS-CoV-2 Proteins In Vitro

Inflammasome activation is one of the first steps in initiating innate immune responses. In this work, we studied the activation of inflammasomes in the airways of critically ill COVID-19 patients and the effects of N-acetylcysteine (NAC) on inflammasomes. Tracheal biopsies were obtained from critically ill patients without COVID-19 and no respiratory disease (control, n = 32), SARS-CoV-2 B.1 variant (n = 31), and B.1.1.7 VOC alpha variant (n = 20) patients. Gene expression and protein expression were measured by RT-qPCR and immunohistochemistry. Macrophages and bronchial epithelial cells were stimulated with different S, E, M, and N SARS-CoV-2 recombinant proteins in the presence or absence of NAC. NLRP3 inflammasome complex was over-expressed and activated in the COVID-19 B.1.1.7 VOC variant and associated with systemic inflammation and 28-day mortality. TLR2/MyD88 and redox NOX4/Nrf2 ratio were also over-expressed in the COVID-19 B.1.1.7 VOC variant. The combination of S-E-M SARS-CoV-2 recombinant proteins increased cytokine release in macrophages and bronchial epithelial cells through the activation of TLR2. NAC inhibited SARS-CoV-2 mosaic (S-E-M)-induced cytokine release and inflammasome activation. In summary, inflammasome is over-activated in severe COVID-19 and increased in B.1.1.7 VOC variant. In addition, NAC can reduce inflammasome activation induced by SARS-CoV-2 in vitro, which may be of potential translational value in COVID-19 patients.     

Comparative study of perovskites as cathode contact materials between an La0.8Sr0.2FeO3 cathode and a Crofer22APU interconnect in solid oxide fuel cells

Perovskites of different compositions were tested as cathode contact material between an La_0.8Sr_0.2FeO₃ cathode and a Crofer22APU interconnect by resistance measurements at 800 °C. The materials tested were LaNi_0.6Fe_0.4O₃ and La_0.8Sr_0.2FeO₃ which are also used as cathodes; La_0.8Sr_0.2Mn_0.5Co_0.5O₃ and La_0.8Sr_0.2Mn_0.1Co_0.3Fe_0.6O₃, selected for comparing perovskites with different Mn contents; and La_0.8Sr_0.2Co_0.75Fe_0.25O₃ and La_0.8Sr_0.2Co_0.75Cu_0.25O₃ for comparing perovskites with high Co content and two possible partial substitutions of the Co. The initial area-specific contact resistance (ASR) was found to depend on the electrical conductivity of the measured perovskites. Time evolution of the ASR depended on the interactions between the contact material and the interconnect, showing the highest degradation rates for LaNi_0.6Fe_0.4O₃ and La_0.8Sr_0.2FeO₃. Chromium from the interconnect reacted with the Sr-containing perovskites forming SrCrO₄. With the contact material without strontium chromium-containing perovskites were formed. A reduced interfacial reaction was achieved by application of a MnCo_1.9Fe_0.1O₄ spinel protection layer on Crofer22APU in terms resulting in low and stable ASR.     

Structural and functional properties of soy protein isolate and cod gelatin blend films

The structure-function relationship of composite films obtained from soybean-protein isolate (SPI) and cod gelatin was studied. Films with different ratios of SPI:gelatin (0, 25, 50, 75, 100% [w/w]) and plasticized by a mixture of glycerol and sorbitol were prepared by casting. Regardless of the soybean-protein concentration, the thickness and water-vapor permeability of the composite films diminished significantly as compared to pure-gelatin films. The formulation containing 25% SPI: 75% cod-skin gelatin had the maximum force at the breaking point, which was 1.8-fold and 2.8-fold greater than those of 100% gelatin and 100% SPI films, respectively. Moreover, this formulation offered high percent-deformation values lower than those of gelatin but higher than all other films containing SPI-, and the same relatively low water-vapor permeability as the 100% SPI film. While all the films exhibited high water solubility, a slight reduction in film solubility and soluble protein was observed with increasing SPI concentration. Differential-scanning calorimetry analyses revealed that gelatin was completely denatured in all films, while soy proteins largely maintained their native conformation. Analysis by fourier-transform–infrared spectroscopy revealed that the presence of 25% SPI produced gelatin conformational changes, self-aggregation of gelatin chains, and intermolecular associations via CO bonds between gelatin and SPI proteins. All films were translucent in appearance, but the yellowish color increased with increasing proportions of the soybean proteins.     

Influence of salt, smoke, and high pressure on growth of Listeria monocytogenes and spoilage microflora in cold-smoked dolphinfish (Coryphaena hippurus)

The effects of different salting and smoking conditions on the growth of Listeria monocytogenes in cold-smoked dolphinfish (Coryphaena hippurus) fillets were evaluated. High concentrations of phenol (72.47 ppm) and salt (3.25%) in muscle inhibited L. monocytogenes growth in smoked fish stored at 20 degrees C for 4 days. The antibacterial effect of high pressure in cold-smoked dolphinfish during long-term chilled (5 degrees C) storage was evaluated in fillets prepared according to two different sets of salting and smoking conditions. Combining the milder salting and smoking conditions (1.97% salt and 42 ppm phenol) with a high pressure treatment of 300 MPa at 20 degrees C for 15 min sufficed to exert a bacteriostatic effect on the total viable bacteria, total lactic acid bacteria, and L. monocytogenes. However, in fillets prepared using the more severe salting and smoking conditions (2.93% salt and 82 ppm phenol), pressurization kept L. monocytogenes counts under the detection limit throughout 100 days of storage. A similar effect was obtained by dosing the fillets with nisin. No luminescent bacteria, hydrogen sulfide-producing bacteria, or Enterobacteriaceae were found in any of the fillets produced using either of the two sets of processing conditions.     

Partial Characterization of Protease Activity in Squid (Todaropsis eblanae) Mantle: Modification by High-pressure Treatment

ABSTRACT: Optimal conditions for proteolytic activity in both nonpressurized and pressurized (300 MPa, 7°C, 20 min) squid ( Todaropsis eblanae ) muscle occurred at acid pH levels (pH 3) over a broad range of temperatures. Pressure treatment did not modify optimal pH and temperatures but did increase proteolytic activity. The acid cysteine proteases, and to a lesser extent the acid serine proteases, were the enzymes mainly affected by the high-pressure treatment. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was indicative of increased protein hydrolysis by pressurization. Myosin heavy chain in both nonpressurized and pressurized squid was degraded at all the temperatures tested, but actin was susceptible only to proteolysis in the pressure-treated muscle at 7°C and 40°C. This behavior was not observed at 55°C.