Evaluation and Characterization of Post-Stroke Lung Damage in a Murine Model of Cerebral Ischemia

After stroke and other brain injuries, there is a high incidence of respiratory complications such as pneumonia or acute lung injury. The molecular mechanisms that drive the brain-lung interaction post-stroke have not yet been elucidated. We performed transient middle cerebral artery occlusion (MCAO) and sham surgery on C57BL/6J mice and collected bronchoalveolar lavage fluid (BALF), serum, brain, and lung homogenate samples 24 h after surgery. A 92 proteins-panel developed by Olink Proteomics^® was used to analyze the content in BALF and lung homogenates. MCAO animals had higher protein concentration levels in BALF than sham-controls, but these levels did not correlate with the infarct volume. No alteration in alveolar-capillary barrier permeability was observed. A total of 12 and 14 proteins were differentially expressed between the groups (FDR < 0.1) in BALF and lung tissue homogenates, respectively. Of those, HGF, TGF-α, and CCL2 were identified as the most relevant to this study. Their protein expression patterns were verified by ELISA. This study confirmed that post-stroke lung damage was not associated with increased lung permeability or cerebral ischemia severity. Furthermore, the dysregulation of HGF, TGF-α, and CCL2 in BALF and lung tissue after ischemia could play an important role in the molecular mechanisms underlying stroke-induced lung damage.     

Alchemical Design of Pharmacological Chaperones with Higher Affinity for Phenylalanine Hydroxylase

Phenylketonuria (PKU) is a rare metabolic disease caused by variations in a human gene, PAH, encoding phenylalanine hydroxylase (PAH), and the enzyme converting the essential amino acid phenylalanine into tyrosine. Many PKU-causing variations compromise the conformational stability of the encoded enzyme, decreasing or abolishing its catalytic activity, and leading to an elevated concentration of phenylalanine in the blood, which is neurotoxic. Several therapeutic approaches have been developed to treat the more severe manifestations of the disorder, but they are either not entirely effective or difficult to adhere to throughout life. In a search for novel pharmacological chaperones to treat PKU, a lead compound was discovered (compound IV) that exhibited promising in vitro and in vivo chaperoning activity on PAH. The structure of the PAH-IV complex has been reported. Here, using alchemical free energy calculations (AFEC) on the structure of the PAH-IV complex, we design a new generation of compound IV-analogues with a higher affinity for the enzyme. Seventeen novel analogues were synthesized, and thermal shift and isothermal titration calorimetry (ITC) assays were performed to experimentally evaluate their stabilizing effect and their affinity for the enzyme. Most of the new derivatives bind to PAH tighter than lead compound IV and induce a greater thermostabilization of the enzyme upon binding. Importantly, the correspondence between the calculated alchemical binding free energies and the experimentally determined ΔΔG_b values is excellent, which supports the use of AFEC to design pharmacological chaperones to treat PKU using the X-ray structure of their complexes with the target PAH enzyme.     

Are electronic white canes better than traditional canes? A comparative study with blind and blindfolded participants

Visually impaired individuals often rely on assistive technologies such as white canes for independent navigation. Many electronic enhancements to the traditional white cane have been proposed. However, only a few of these proof-of-concept technologies have been tested with authentic users, as most studies rely on blindfolded non-visually impaired participants or no testing with participants at all. Experiments involving blind users are usually not contrasted with the traditional white cane. This study set out to compare an ultrasound-based electronic cane with a traditional white cane. Moreover, we also compared the performance of a group of visually impaired participants (N = 10) with a group of blindfolded participants without visual impairments (N = 31). The results show that walking speed with the electronic cane is significantly slower compared to the traditional white cane. Moreover, the results show that the performance of the participants without visual impairments is significantly slower than for the visually impaired participants. No significant differences in obstacle detection rates were observed across participant groups and device types for obstacles on the ground, while 79% of the hanging obstacles were detected by the electronic cane. The results of this study thus suggest that electronic canes present only one advantage over the traditional cane, namely in its ability to detect hanging obstacles, at least without prolonged practice. Next, blindfolded participants are insufficient substitutes for blind participants who are expert cane users. The implication of this study is that research into digital white cane enhancements should include blind participants. These participants should be followed over time in longitudinal experiments to document if practice will lead to improvements that surpass the performance achieved with traditional canes.

     

Thiophene‐ and silarylene‐containing polyesters. Resonance effect on conductivity after polarization in an external electric field

Polyesters were synthesized by direct polycondensation of thiophene‐2,5‐dicarboxylic acid and five different silarylene‐containing diphenols using a tosyl chloride/pyridine/N,N‐dimethylformamide system as a condensing agent. Polymers were obtained in good yields and were characterized using Fourier transform infrared and NMR (¹H, ¹³C, 135‐DEPT and ²⁹Si) spectroscopy and elemental analysis. All polymers were completely soluble in aprotic organic polar solvents such as dimethylformamide, dimethylsulfoxide and N‐methyl‐2‐pyrrolidone. The range of effective mass of the polymers (m/z) was 1 × 10⁵–2 × 10⁵, determined using electrospray ionization mass spectrometry. Asymmetry and steric hindrance prevented dense packing of the polymeric chains, showing glass transition temperatures between ? 78 and ? 51 °C and loss of thermal stability at 177–199 °C (10% weight loss). Additionally, the melting points of the polyesters were found to be in the range 62–67 °C. Because of this, the samples were semi‐solid at room temperature. The optical band gaps of the polymers were observed between 4.54 and 4.48 eV, corresponding in all cases to insulator behavior. The molecular structure of the samples was studied using X‐ray diffraction, showing a degree of order that was associated with two monoclinic lattices. Additionally, the conductivity was studied using a two‐point method with contacts on top of polymer films. Prior to the electrical measurement, the samples were polarized in an external electric field of 0.8 to 6.4 V cm^?1, and the alignment of the dipoles increased the electrical conductivity. Copyright © 2012 Society of Chemical Industry     

Capillary electrophoretic profiling of tryptic digests of water soluble proteins from Bacillus thuringiensis-transgenic and non-transgenic maize species

Capillary zone electrophoresis (CZE) has been applied to separation and characterisation of enzymatic (tryptic) hydrolysates of water-soluble proteins from Bacillus thuringiensis (Bt)-transgenic (Aristis-Bt) and two native non-transgenic (Aristis and Coventry) maize varieties. Water-soluble proteins were extracted from the flour of these maize species and digested by bovine pancreatic trypsin immobilised on agarose gel in 100 mM ammonium hydrocarbonate buffer, pH 7.9. The yielded tryptic digests of proteins were analysed by CZE in four acidic background electrolytes (BGEs) (100 mM H₃PO₄, 50 mM Tris, pH 2.25; 500 mM acetic acid, pH 2.54; 200 mM formic acid, 200 mM acetic acid, pH 2.05; and 200 mM iminodiacetic acid, pH 2.26) using a lab-made CZE apparatus equipped with bare fused silica capillary and UV-absorption detector operating at 206 nm. Among the tested BGEs, the best resolution of the tryptic peptides of extracted proteins of the above three maize species was obtained in isoelectric BGE, 200 mM iminodiacetic acid, pH 2.26. Selected resolved tryptic peptides of proteins were characterised by effective electrophoretic mobilities and corrected (migration times normalised) peak areas. Some significant relative qualitative and quantitative differences in CZE-UV profiling of tryptic protein digests were found, which can be potentially used to differentiate transgenic Aristis Bt and non-transgenic Aristis varieties or two native non-transgenic varieties, Aristis and Coventry.     

A study on the effect of the asymmetry on real capacitated vehicle routing problems

Matrices with distances between pairs of locations are essential for solving vehicle routing problems like the Capacitated Vehicle Routing Problem (CVRP), Traveling Salesman Problem (TSP) and others. This work deals with the complex reality of transportation networks and asymmetry. Through a series of comprehensive and thorough computational and statistical experiments we study the effect that many factors like asymmetry, geographical location of the depot and clients, demand, territory and maximum vehicle capacity have in the solution of CVRP instances. We examine both classical heuristics as well as current state-of-the-art metaheuristics and show that these methods are seriously affected by the studied factors from a solution time and quality of solutions perspective. We systematically compare the solutions obtained in the symmetric scenario with those obtained in the real asymmetric case at a quantitative as well as a qualitative level, with the objective of carefully measuring and understanding the differences between both cases.     

Comprehensive characterization of the functional activities of pressurized liquid and ultrasound-assisted extracts from Chlorella vulgaris

Chlorella vulgaris has been referred as a potential source of bioactive compounds (carotenoids and fatty acids). In this work, the ability of an environmentally friendly extraction technique such as Pressurized Liquid Extraction (PLE) and a traditional extraction technique such as Ultrasound-Assisted Extraction (UAE) to obtain functional compounds from C. vulgaris, at analytical scale, has been demonstrated. Seeked bioactivities were antioxidant and antimicrobial, for their interest in the food industry. Therefore, a methodology including analytical extraction, in-vitro assays and chemical characterization via HPLC-DAD and GC–MS has been used to determine the interest of Chlorella as a source of functional food ingredients. Results demonstrated that PLE provide higher yields than UAE while similar bioactivities were obtained. Important concentration of carotenoids (lutein, neoxanthin, β-carotene, etc.), chlorophylls, sterols, phytols, and fatty acids (among others) have been found in Chlorella extracts that could be correlated to the observed biological activity.     

Online Scheduling FIFO Policies with Admission and Push-Out

We consider the problem of managing a bounded size First-In-First-Out (FIFO) queue buffer, where each incoming unit-sized packet requires several rounds of processing before it can be transmitted out. Our objective is to maximize the total number of successfully transmitted packets. We consider both push-out (when a policy is permitted to drop already admitted packets) and non-push-out cases. We provide worst-case guarantees for the throughput performance of our algorithms, proving both lower and upper bounds on their competitive ratio against the optimal algorithm, and conduct a comprehensive simulation study that experimentally validates predicted theoretical behavior.