Role of Tyrosine Kinase Syk in Thrombus Stabilisation at High Shear

Understanding the pathways involved in the formation and stability of the core and shell regions of a platelet-rich arterial thrombus may result in new ways to treat arterial thrombosis. The distinguishing feature between these two regions is the absence of fibrin in the shell which indicates that in vitro flow-based assays over thrombogenic surfaces, in the absence of coagulation, can be used to resemble this region. In this study, we have investigated the contribution of Syk tyrosine kinase in the stability of platelet aggregates (or thrombi) formed on collagen or atherosclerotic plaque homogenate at arterial shear (1000 s⁻¹). We show that post-perfusion of the Syk inhibitor PRT-060318 over preformed thrombi on both surfaces enhances thrombus breakdown and platelet detachment. The resulting loss of thrombus stability led to a reduction in thrombus contractile score which could be detected as early as 3 min after perfusion of the Syk inhibitor. A similar loss of thrombus stability was observed with ticagrelor and indomethacin, inhibitors of platelet adenosine diphosphate (ADP) receptor and thromboxane A₂ (TxA₂), respectively, and in the presence of the Src inhibitor, dasatinib. In contrast, the Btk inhibitor, ibrutinib, causes only a minor decrease in thrombus contractile score. Weak thrombus breakdown is also seen with the blocking GPVI nanobody, Nb21, which indicates, at best, a minor contribution of collagen to the stability of the platelet aggregate. These results show that Syk regulates thrombus stability in the absence of fibrin in human platelets under flow and provide evidence that this involves pathways additional to activation of GPVI by collagen.     

Label-Free Study of the Global Cell Behavior during Exposure to Environmental Radiofrequency Fields in the Presence or Absence of Pro-Apoptotic or Pro-Autophagic Treatments

It remains controversial whether exposure to environmental radiofrequency signals (RF) impacts cell status or response to cellular stress such as apoptosis or autophagy. We used two label-free techniques, cellular impedancemetry and Digital Holographic Microscopy (DHM), to assess the overall cellular response during RF exposure alone, or during co-exposure to RF and chemical treatments known to induce either apoptosis or autophagy. Two human cell lines (SH-SY5Y and HCT116) and two cultures of primary rat cortex cells (astrocytes and co-culture of neurons and glial cells) were exposed to RF using an 1800 MHz carrier wave modulated with various environmental signals (GSM: Global System for Mobile Communications, 2G signal), UMTS (Universal Mobile Telecommunications System, 3G signal), LTE (Long-Term Evolution, 4G signal, and Wi-Fi) or unmodulated RF (continuous wave, CW). The specific absorption rates (S.A.R.) used were 1.5 and 6 W/kg during DHM experiments and ranged from 5 to 24 W/kg during the recording of cellular impedance. Cells were continuously exposed for three to five consecutive days while the temporal phenotypic signature of cells behavior was recorded at constant temperature. Statistical analysis of the results does not indicate that RF-EMF exposure impacted the global behavior of healthy, apoptotic, or autophagic cells, even at S.A.R. levels higher than the guidelines, provided that the temperature was kept constant.     

Analysis of Integrin αIIb Subunit Dynamics Reveals Long-Range Effects of Missense Mutations on Calf Domains

Integrin α_IIbβ₃, a glycoprotein complex expressed at the platelet surface, is involved in platelet aggregation and contributes to primary haemostasis. Several integrin α_IIbβ₃ polymorphisms prevent the aggregation that causes haemorrhagic syndromes, such as Glanzmann thrombasthenia (GT). Access to 3D structure allows understanding the structural effects of polymorphisms related to GT. In a previous analysis using Molecular Dynamics (MD) simulations of α_IIb Calf-1 domain structure, it was observed that GT associated with single amino acid variation affects distant loops, but not the mutated position. In this study, experiments are extended to Calf-1, Thigh, and Calf-2 domains. Two loops in Calf-2 are unstructured and therefore are modelled expertly using biophysical restraints. Surprisingly, MD revealed the presence of rigid zones in these loops. Detailed analysis with structural alphabet, the Proteins Blocks (PBs), allowed observing local changes in highly flexible regions. The variant P741R located at C-terminal of Calf-1 revealed that the Calf-2 presence did not affect the results obtained with isolated Calf-1 domain. Simulations for Calf-1 + Calf-2, and Thigh + Calf-1 variant systems are designed to comprehend the impact of five single amino acid variations in these domains. Distant conformational changes are observed, thus highlighting the potential role of allostery in the structural basis of GT.     

Recovery of vinyl chloride from by-streams of polyvinyl chloride production by TPSA in a multitubular adsorber

This work aims at developing an efficient and feasible adsorption-based separation process for the separation of vinyl chloride and nitrogen, on activated carbon, by employing a multitubular packed bed geometry, with adsorbent material inside the tubes. Using this geometry, a 2-dimensional mathematical model of a temperature pressure swing adsorption process was used to developed a 6-step three multitubular adsorbers system capable of separating and purifying an industrial scale gas stream of a 40:60% (v/v) vinyl chloride/nitrogen mixture into a 95% (v/v) vinyl chloride stream and a nitrogen stream with a vinyl chloride limit concentration of 8 ppm (w/w). The process reported energy consumption of 4.88 × 10⁶ J/kg_VCM and recovery capacity of 24.35 kg_VCM/(m³_unit h). The multitubular geometry enabled the use of lower adsorbent loads, shorter cycle times, and lower regeneration temperatures. An equivalent 1-dimensional model has also shown to satisfactorily estimate the performance of the current equipment.     

Synthesis and Characterization of Ti-Nb2O5 Catalysts for Discoloration Reaction of Bromophenol Blue and Indigo Carmine Dyes

Topics in Catalysis - This work investigated the photodegradation of bromophenol blue (BPB) and indigo carmine (IC) dye under UV–Visible light using pure oxides of titanium and niobium, as...     

Active application of primer acid on acid‐treated enamel: Influence on the bond effectiveness of self‐etch adhesives systems

Aim: Evaluate the composite‐to‐enamel bond after passive or active application of self‐etching primer systems on polished or pre‐etched enamel with phosphoric acid. Materials and Methods: Two self‐etch adhesives systems (SEAS) were used: Clearfil SE Bond and Easy Bond. Third human molars were divided into 8 groups (N = 10). The crown of each tooth was sectioned into halves and the mesial/distal surfaces were used. The adhesives were actively or passively applied on enamel with or without prior phosphoric‐acid etching. Resin composite cylinders were built after adhesive application. After stored in relative humidity for 24 hr/37°C the specimens were subjected to microshear test in universal testing a machine at a crosshead speed of 0.5 mm/minute. The results were analyzed with three‐way ANOVA and the Tukey test. The enamel‐etching pattern was evaluated under SEM. Results: The 2‐step SEAS system presented significantly higher adhesive bond strength means (47.37 MPa) than the 1‐step (36.87 MPa). A poor enamel‐ etching pattern was observed in active mode showing irregular and short resin tags, however there was not compromised the bond strength. Conclusion: Active or passive application produced similar values of bond strength to enamel regardless of enamel pretreatment and type of SEAS.     

Influence of chemical structure of hardener on mechanical and adhesive properties of epoxy polymers

The mechanical and adhesive properties of epoxy formulations based on diglycidyl ether of bisphenol A cured with various aliphatic amines were evaluated in the glass state. Impact and uniaxial compression tests were used to determine the impact energy, elastic modulus and yield stress, respectively. The adhesion tests were carried out in steel–steel joints using single‐lap shear, T‐peel, and impact adhesive joints geometry. The better mechanical and adhesive behavior of the networks is obtained when exists high flexibility of chain between crosslink and/or high elastic modulus. The 1‐(2‐aminoethyl)piperazine epoxy network presents the best adhesive properties, high flexibility, and the largest impact energy. However, it possesses low elastic modulus and yield stress. Also, exhibits increases in peel strength and impact energy while reductions in lap shear strength. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Thermovinification of grapes from the Cabernet Sauvignon and Pinot Noir varieties using immobilized yeasts

This study examined the thermovinification technique using yeasts immobilized on calcium alginate for microvinification of the Cabernet Sauvignon and Pinot Noir grape varieties grown in Diamantina, Minas Gerais, Brazil. The wines obtained by thermovinification were characterized by their chemical and sensory aspects. The varietal wines from Cabernet Sauvignon and Pinot Noir had an alcohol content of 9.3 and 10.9 % (v/v), respectively. The total acidity, volatile acidity and the alcohol/reduced dry extract ratio were consistent with the Brazilian standards established for identity and quality of wines. There was a significant reduction in the levels of anthocyanins, flavonoids and total phenolic compounds after aging in the bottle. The sensory evaluation presented an acceptability exceeding 60 % for both wines.     

Microbiological characterization of randomly selected Portuguese raw milk cheeses with reference to food safety

Seventy raw milk cheeses made in different regions of Portugal, both hard and soft varieties, made with cow's, ewe's, or goat's milk or combinations of these, were sampled within their quoted shelf lives for microbiological safety. On the basis of the presence or numbers of Escherichia coli, E. coli O157, Staphylococcus aureus, Salmonella, and Listeria monocytogenes, cheeses were categorized as satisfactory, acceptable, unsatisfactory, or unacceptable and potentially hazardous. Twenty-two of the 70 cheeses were classified as satisfactory or acceptable. Thirty-seven of the cheeses were considered unsatisfactory because of the presence of E. coli, S. aureus, or both, while 11 of the cheeses were graded as unacceptable and potentially hazardous because of the presence of excessive numbers of S. aureus, E. coli, or L. monocytogenes and the presence of Salmonella in three of these. All cheeses graded as unacceptable and potentially hazardous were soft or semisoft cheeses made with ewe's and goat's milk, with the exception of two hard cheeses made with cow's milk. E. coli O157 was not detected in any of the cheeses. According to the present results, it seems that the presence or counts of pathogenic or indicator organisms in raw milk cheeses cannot be related to the processing conditions, milk type, or region of production.