The Emergence of New Catalytic Abilities in an Endoxylanase from Family GH10 by Removing an Intrinsically Disordered Region

Endoxylanases belonging to family 10 of the glycoside hydrolases (GH10) are versatile in the use of different substrates. Thus, an understanding of the molecular mechanisms underlying substrate specificities could be very useful in the engineering of GH10 endoxylanases for biotechnological purposes. Herein, we analyzed XynA, an endoxylanase that contains a (β/α)₈-barrel domain and an intrinsically disordered region (IDR) of 29 amino acids at its amino end. Enzyme activity assays revealed that the elimination of the IDR resulted in a mutant enzyme (XynAΔ29) in which two new activities emerged: the ability to release xylose from xylan, and the ability to hydrolyze p-nitrophenyl-β-d-xylopyranoside (pNPXyl), a substrate that wild-type enzyme cannot hydrolyze. Circular dichroism and tryptophan fluorescence quenching by acrylamide showed changes in secondary structure and increased flexibility of XynAΔ29. Molecular dynamics simulations revealed that the emergence of the pNPXyl-hydrolyzing activity correlated with a dynamic behavior not previously observed in GH10 endoxylanases: a hinge-bending motion of two symmetric regions within the (β/α)₈-barrel domain, whose hinge point is the active cleft. The hinge-bending motion is more intense in XynAΔ29 than in XynA and promotes the formation of a wider active site that allows the accommodation and hydrolysis of pNPXyl. Our results open new avenues for the study of the relationship between IDRs, dynamics and activity of endoxylanases, and other enzymes containing (β/α)₈-barrel domain.     

Maitotoxin, a cationic channel activator

ICAM-1 mediates the recruitment of neutrophils through the endothelium to the site of inflammation by the ICAM-1/Mac-1 and ICAM-1/LFA-1 adhesion pathways. In extrahepatic cholestasis, recruitment of neutrophils is a main feature of the inflammatory infiltrate in areas of parenchymal damage. The aim of the present study was to describe the light and electron microscopical localization of ICAM-1 expression in the liver of cholestatic patients. The peroxidase-antiperoxidase technique was used. Increased ICAM-1 expression was detected on sinusoidal endothelial and Kupffer cells. A de novo ICAM-1 expression was described on some Ito cells and the sinusoidal hepatocyte membrane in areas of parenchymal injury. In the portal areas of livers of cholestatic patients, ICAM-1 was observed on the endothelial surface of portal veins and on hepatic arteries. Occasionally, ICAM-1 was found on the surface of bile duct epithelia. It is suggested that ICAM-1 expression is up-regulated by cytokines like TNF-alpha, IL-1 and interferons released from activated Kupffer cells. The mechanisms of ICAM-1 upregulation and neutrophil recruitment in the liver during extrahepatic cholestasis are discussed.     

Centrifugally spun mats based on biopolyesters/hydroxyapatite and their potential as bone scaffolds

This work studied the potential of centrifugal spinning for the production of fibrous materials based on poly(D,L-lactic acid) (PDLLA) and poly(3-hydroxybutyrate) (PHB) with hydroxyapatite nanoparticles (n-Hap). The influence of n-Hap concentration (5, 10, and 15 wt.%) and spinneret angular speed on the final fiber morphology were analyzed. Further experimental evaluations were implemented to determine the effect of n-Hap on the thermal and mechanical performance. The optimum parameters that show a balance among high yield production of homogeneous fibers with the smallest fiber average diameter were found to be at 5 wt.% of n-Hap processed at 7000 rpm for PDLLA, and 5, 10, and 15 wt.% of n-Hap at 6000 rpm for PHB. The thermal stability, for both systems, was not significantly affected. The mechanical performance of PHB systems was improved with the addition of n-Hap. Osteoblast cell viability tests depicted a favorable cell response on the PDLLA systems.     

Detecting dynamic system regime boundaries with Fisher information: the case of ecosystems

Clean Technologies and Environmental Policy - The direct measurement of the resilience (resistance to disturbances) of an ecosystem’s current regime (or “alternative stable...     

Body Sensations Associated With Emotions in Rarámuri Indians, Rural Javanese, and Three Student Samples.

Cultural variations in the associations of 12 body sensations with 7 emotions were studied in 2 rural samples from northern Mexico (n = 61) and Java, Indonesia (n = 99), with low exposure to Western influences and in 3 university student samples from Belgium (n = 75), Indonesia (n = 85), and Mexico (n = 123). Both parametric and nonparametric analyses suggest that findings from previous studies with only student samples (K. R. Scherer & H. G. Wallbott, 1994) were generalizable to the 2 rural samples. Some notable cultural deviations from common profiles were also identified. Implications of the findings for explanations of body sensations experienced with emotions and the cross-cultural study of emotions are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effect of consanguinity on Argentinean Angus beef DNA traceability

Since the 1990s several authors have envisaged the use of DNA to certify meat origin. Two major parameters must be assessed before a DNA based traceability protocol can be implemented in the food chain: (i) the information content of a DNA marker set in a specific livestock breed or group of breeds; (ii) the minimum number of DNA markers needed to obtain a statistically acceptable match probability. The objective of the present work was to establish the effect of different levels of inbreeding in the matching efficiency, and the minimum number of microsatellite markers needed, in a DNA based meat traceability program, starting from an 11-microsatellite marker panel. Samples were obtained from beef production farms in South America, where animals are typically bred under pasture-based extensive conditions. Three groups of animals with different consanguinity rates were sampled. Exclusion power (Q) was higher than 0.999998 and match probability lower than 3.01E−08, for the whole set of markers within each group. Both values were affected by consanguinity. To reach a two mismatch criteria exclusion power (Q₂) of 99.99, six markers were needed in unrelated animals whereas seven markers were needed in related animals. To reach Q₂ = 99.9999, 8 and 10 microsatellite markers, respectively, were needed. In general, one or two more microsatellite markers were needed to identify consanguineous animals. This study proved the DNA marker set used to be suitable for the identification of the meat from all slaughtered animals in Argentina, per week, month, and year.     

In Silico Drug Repositioning to Target the SARS-CoV-2 Main Protease as Covalent Inhibitors Employing a Combined Structure-Based Virtual Screening Strategy of Pharmacophore Models and Covalent Docking

The epidemic caused by the SARS-CoV-2 coronavirus, which has spread rapidly throughout the world, requires urgent and effective treatments considering that the appearance of viral variants limits the efficacy of vaccines. The main protease of SARS-CoV-2 (M^pro) is a highly conserved cysteine proteinase, fundamental for the replication of the coronavirus and with a specific cleavage mechanism that positions it as an attractive therapeutic target for the proposal of irreversible inhibitors. A structure-based strategy combining 3D pharmacophoric modeling, virtual screening, and covalent docking was employed to identify the interactions required for molecular recognition, as well as the spatial orientation of the electrophilic warhead, of various drugs, to achieve a covalent interaction with Cys145 of M^pro. The virtual screening on the structure-based pharmacophoric map of the SARS-CoV-2 M^pro in complex with an inhibitor N3 (reference compound) provided high efficiency by identifying 53 drugs (FDA and DrugBank databases) with probabilities of covalent binding, including N3 (Michael acceptor) and others with a variety of electrophilic warheads. Adding the energy contributions of affinity for non-covalent and covalent docking, 16 promising drugs were obtained. Our findings suggest that the FDA-approved drugs Vaborbactam, Cimetidine, Ixazomib, Scopolamine, and Bicalutamide, as well as the other investigational peptide-like drugs (DB04234, DB03456, DB07224, DB7252, and CMX-2043) are potential covalent inhibitors of SARS-CoV-2 M^pro.     

Determination of DNA adducts of malonaldehyde in humans: effects of dietary fatty acid composition

The effects of dietary fatty acid composition on the endogenous formation of DNA adducts of malonaldehyde (MA), the major product of lipid peroxidation, were investigated in humans. A group of 59 healthy individuals of both sexes and different ages was initially fed a milk fat-based diet rich in saturated fatty acids for 14 days. Following this initial period, after which the group was considered homogeneous with respect to diet, 30 randomly chosen subjects were given a sunflower oil-based (rich in polyunsaturated fatty acids) (SO) diet and the remaining 29 individuals a low erucic acid rapeseed oil-based (rich in monounsaturated fatty acids) (RO) diet for 25 days. The fatty acid composition of plasma lipid fractions and the level of DNA adducts of MA in total white blood cells were then determined at the end of the SO and RO dietary periods. DNA adduct levels were measured by 32p-postlabelling using reversed-phase HPLC with on-line detection of radioactivity. Higher concentrations of polyunsaturated fatty acids in plasma triglycerides and higher levels of DNA adducts of MA were found in the subjects on the SO diet when compared with those in the RO dietary group. A large inter-individual variation in adduct levels was observed. The average adduct level in the SO diet group was 7.4 +/- 8.7 adducts/10(7) nucleotides (n = 23). This level was 3.6-fold higher than that found in individuals in the RO diet group (P < 0.001). Our results, in conjunction with the mutagenic and carcinogenic properties of MA, thus suggest the interaction of lipid peroxidation products such as MA with DNA as one plausible mechanism explaining the involvement of dietary fat in carcinogenesis.     

Microstructural Development of ZnO Pellets Doped with Different Vanadium Oxides (V2O5 and V2O3)

This work presents the effect of zinc oxide (ZnO) ceramics, doped with different percentages of vanadium trioxide (V₂O₃) and vanadium pentoxide (V₂O₅). Samples were analyzed by X-ray diffraction and scanning electron microscopy. The addition of V₂O₃ or V₂O₅ produced changes in the composition and morphology of the pellets. In both cases, different zinc vanadates were detected as secondary phases: α-Zn₃(VO₄)₂ and Zn₄V₂O₉. Furthermore, when the vanadium concentration was equal to or higher than 3 wt%, the presence of filaments was detected on the surface of the pellets. These filaments were produced due to vanadium segregation. However, this effect was only observed at the surface of the pellets. On the bulk, the filaments were not observed. Instead, vanadium was found at the interfaces between the ZnO grains and at triple points, as it could be expected.