Human cerebrovascular contractile receptors are upregulated via a B-Raf/MEK/ERK-sensitive signaling pathway

Background  

Cerebral ischemia results in a rapid increase in contractile cerebrovascular receptors, such as the 5-hydroxytryptamine type 1B (5-HT_1B), angiotensin II type 1 (AT₁), and endothelin type B (ET_B) receptors, in the vessel walls within the ischemic region, which further impairs local blood flow and aggravates tissue damage. This receptor upregulation occurs via activation of the mitogen-activated protein kinase pathway. We therefore hypothesized an important role for B-Raf, the first signaling molecule in the pathway. To test our hypothesis, human cerebral arteries were incubated at 37°C for 48 h in the absence or presence of a B-Raf inhibitor: SB-386023 or SB-590885. Contractile properties were evaluated in a myograph and protein expression of the individual receptors and activated phosphorylated B-Raf (p-B-Raf) was evaluated immunohistochemically.     

Site-Specific Hyperphosphorylation Inhibits,Rather than Promotes,Tau Fibrillization,Seeding Capacity,and Its Microtubule Binding

The consistent observation of phosphorylated tau in the pathology of Alzheimer's disease has contributed to the emergence of a model where hyperphosphorylation triggers both tau disassociation from microtubules and its subsequent aggregation. Herein, we applied a total chemical synthetic approach to site-specifically phosphorylate the microtubule binding repeat domain of tau (K18) at single (pS356) or multiple (pS356/pS262 and pS356/pS262/pS258) residues. We show that hyperphosphorylation of K18 inhibits 1) its aggregation in vitro, 2) its seeding activity in cells, 3) its binding to microtubules, and 4) its ability to promote microtubule polymerization. The inhibition increased with increasing the number of phosphorylated sites, with phosphorylation at S262 having the strongest effect. Our results argue against the hyperphosphorylation hypothesis and underscore the importance of revisiting the role of site-specific hyperphosphorylation in regulating tau functions in health and disease.     

Insight into serum protein interactions with functionalized magnetic nanoparticles in biological media

Surface modification with linear polymethacrylic acid (20 kDa), linear and branched polyethylenimine (25 kDa), and branched oligoethylenimine (800 Da) is commonly used to improve the function of magnetite nanoparticles (MNPs) in many biomedical applications. These polymers were shown herein to have different adsorption capacity and anticipated conformations on the surface of MNPs due to differences in their functional groups, architectures, and molecular weight. This in turn affects the interaction of MNPs surfaces with biological serum proteins (fetal bovine serum). MNPs coated with 25 kDa branched polyethylenimine were found to attract the highest amount of serum protein while MNPs coated with 20 kDa linear polymethacrylic acid adsorbed the least. The type and amount of protein adsorbed, and the surface conformation of the polymer was shown to affect the size stability of the MNPs in a model biological media (RPMI-1640). A moderate reduction in r(2) relaxivity was also observed for MNPs suspended in RPMI-1640 containing serum protein compared to the same particles suspended in water. However, the relaxivities following protein adsorption are still relatively high making the use of these polymer-coated MNPs as Magnetic Resonance Imaging (MRI) contrast agents feasible. This work shows that through judicious selection of functionalization polymers and elucidation of the factors governing the stabilization mechanism, the design of nanoparticles for applications in biologically relevant conditions can be improved.     

Application of factorial designs and Doehlert matrix in optimization of experimental variables associated with the preconcentration and determination of vanadium and copper in seawater by inductively coupled plasma optical emission spectrometry

In the present paper a procedure for preconcentration and determination of vanadium and copper in seawater using inductively coupled plasma optical emission spectrometry (ICP OES) is proposed, which is based on solid-phase extraction of vanadium (IV), vanadium (V) and copper (II) ions as 1-(2-pyridylazo)-2-naphthol (PAN) complexes by active carbon. The optimization process was carried out using two-level full factorials and Doehlert matrix designs. Four variables (PAN mass, pH, active carbon mass and shaking time) were regarded as factors in the optimization. Results of the two-level full factorial design 2⁴ with 16 runs for vanadium extraction, based on the variance analysis (ANOVA), demonstrated that the factors pH and active carbon mass, besides the interaction (pH×active carbon mass), are statistically significant. For copper, the ANOVA revealed that the factors PAN mass, pH and active carbon mass and the interactions (PAN mass×pH) and (pH×active carbon mass) are statistically significant. Doehlert designs were applied in order to determine the optimum conditions for extraction. The procedure proposed allowed the determination of vanadium and copper with detection limits (3σ/S) of 73 and 94 ng l⁻¹, respectively. The precision, calculated as relative standard deviation (R.S.D.), was 1.22 and 1.37% for 12.50 μg l⁻¹ of vanadium and copper, respectively. The preconcentration factor was 80. The recovery achieved for determination of vanadium and copper in the presence of several cations demonstrated that this procedure improved the selectivity required for seawater analysis. The procedure was applied to the determination of vanadium and copper in seawater samples collected in Salvador City, Brazil. Results showed good agreement with other data reported in the literature.     

Synchronization structure of evolving epileptic networks using cross-entropy

In this paper we present connectivity patterns of evolving large scale epileptic networks. We employed a cross-entropy measure in the frequency domain on EEG signals to infer the networks, before and during episodes of epileptic seizures. This measure allowed us to make a richer portrait about the node interactions on the graph and to identify emergent structures associated with the synchronization of brain activity. Our results points to a more complex scenario of network organization than the synchronized/unsynchronized dichotomy, with two main results: first, showing regions with unsynchronized (or independent) behavior, even during absence seizures, contradicting the concept of hypersynchrony. Furthermore, we explore the cross-entropy fluctuations along the seizure: a group of nodes became more similar over time while another group became more different, showing a complementary behaviour and different local brain activities. These results bring new questions about the spreading and the sustenance of the epileptic seizures and others synchronization phenomena in living systems.     

Molybdenum determination in iron matrices by ICP-AES after separation and preconcentration using polyurethane foam

A procedure is proposed for the separation and determination of molybdenum in iron matrices by a batch process. It is based on the solid-phase extraction of the molybdenum(V) ion as thiocyanate complex on polyurethane (PU) foam. The extraction parameters were optimized. Using 0.20 mol L^–1 hydrochloric acid, a thiocyanate concentration of 0.10 mol L^–1, 100 mg of polyurethane foam and shaking time of 10 min, molybdenum (5–400 μg) can be separated and preconcentrated from large amounts of iron (10 mg). Desorption was carried out instantaneously by conc. nitric acid or acetone. Distribution coefficients, sorption capacity of the PU foam and coefficients of variation were also evaluated. The effect of some ions on the separation procedure was assessed. Iron(III) should be reduced to iron(II). The proposed procedure was used to determine molybdenum in standard iron matrices such as steel and pure iron. The achieved results did not show significant differences with certified values.     

Kinetics of simultaneous decrystallization and crystallization induced in high-modulus rayon by bromine water

Studies of the effect of aqueous Br₂ solution on the fine structure of cellulose have shown that the accessibility of the cellulose is a complex function of the duration of treatment. Following immersion of rayon-cellulose in the Br₂ solution, its accessibility is observed to increase until a peak is reached, after which the accessibility decreases with time. For higher temperatures and greater concentrations of Br₂, a sharper peak and faster rates of increase and decrease of accessibility are obtained. In order to explain this behavior, a mathematical model of the kinetics is postulated involving two opposing first-order processes occurring simultaneously. One process describes the decrystallization of crystalline regions within the cellulose by opening intermediate linkages due to Van-der-Waals forces involved. The second process relates to the crystallization of the less-ordered regions. The rate constants for these processes are determined by fitting experimental curves to the postulated analytical expression with the aid of a digital computer program. Experiments carried out on high-modulus rayon showed that for this type of cellulose, the effect described above was very pronounced. From the calculated rate constants, the activation energies of the crystallization and decrystallization reactions were determined. The plots of the rate constants versus 1/T showed that a break occurs at 25°C, corresponding to a second-order transition point of cellulose. The energies of activation of the crystallization process are in the range of 7–9 and 10–13 kcal/mol in the temperature ranges 10–25 and 25–40°C, respectively. The activation energies of the decrystallization process are 13 and 18–24 kcal/mol in these temperature ranges.     

Electroconductive Green Metal-polyaniline Nanocomposites: Synthesis and Application in Sensors

Polyaniline (PANI) is one of the most extensively used conducting polymer due to its fascinating properties including conducting, thermal, optical, magnetic and electrochemical properties, simple synthesis procedure and low cost of monomer. It has attracted major attention in a variety of applications including electrochemical sensors, catalysts, supercapacitors and biosensors. However, its limitations such as insolubility in common solvents, low process-ability and poor mechanical properties have led to the development of new approaches to improve it properties. Metal nanoparticles (MNPs) such as silver, gold, copper and palladium have been combined with PANI to improve on its properties which has led to a new class of materials known as metal/PANI nanocomposites. These hybrid nanocomposites incorporate advantages of both MNPs and polymers which effectively improves the properties of the individual materials. Various synthesis techniques including in situ polymerization, ɤ-radiolysis, electrodeposition, complexation, vacuum deposition and interfacial polymerization have been used in the formation of metal/PANI nanocomposites. These nanocomposites have been used in various sensor and biosensor applications due to their excellent conductivity, ease of synthesis, excellent redox potentials, chemical and thermal stability. This review highlights the various metal/PANI nanocomposites, their various synthesis techniques and their application in sensors and biosensors. The importance of these nanocomposites in sensing and signaling various toxic heavy metals such as mercury, lead and silver and toxic gases such as hydrogen sulphide, ammonia and chloroform has been discussed. In addition the review covers the applications of metal/PANI nanocomposites in biosensor systems for the detection of glucose, DNA, protein, cholesterol, drugs and hydrogen peroxide.