New approach to immobilization and specific-sequence detection of nucleic acids based on poly(4-hydroxyphenylacetic acid)

Immobilization and hybridization of oligonucleotides or specific-gene PCR product (DENV-1), a conserved genomic sequence of the dengue virus, onto graphite electrode modified with poly(4-hydroxyphenylacetic acid), were carried out with success using both direct electrochemical oxidation of guanine or redox electroactive indicator ethidium bromide.Studies of oligonucleotides hybridization with the complementary target showed a decrease of both guanosine and adenosine current peaks, when compared with the peak previously obtained before the hybridization. Immobilized ssDNA, DENV-1, was hybridized with various concentrations of target DNA. The interaction between DENV-1 hybridized onto the modified graphite electrodes surface and the intercalator, ethidium bromide, was observed by differential pulse voltammetry, monitoring the current change generated to the DNA intercalator accumulated onto the modified electrode after DNA hybridization. For the determination of complementary target, the proposed method exhibited a good dynamic range (12–42 nmol L^? 1) and a low detection limit (7.12 nmol L^? 1).AFM images showed that the oligonucleotides or single-stranded DNA, DENV-1, before hybridization, had roughness values lower than the double stranded obtained after hybridization.The new surface obtained in these work, as well as the possibility of utilization of the same to monitor hybridization events is a promising strategy for the development of DNA electrochemical biosensors.     

Molecular Dynamics of CYFIP2 Protein and Its R87C Variant Related to Early Infantile Epileptic Encephalopathy

The CYFIP2 protein (cytoplasmic FMR1-interacting protein 2) is part of the WAVE regulatory complex (WRC). CYFIP2 was recently correlated to neurological disorders by the association of the R87C variant with early infantile epileptic encephalopathy (EIEE) patients. In this set of syndromes, the epileptic spasms and seizures since early childhood lead to impaired neurological development in children. Inside the WRC, the variant residue is at the CYFIP2 and WAVE1 protein interface. Thus, the hypothesis is that the R87C modification weakens this interaction, allowing the WRC complex’s constant activation. This work aimed to investigate the impacts of the mutation on the structure of the WRC complex through molecular dynamics simulation. For that, we constructed WRC models containing WAVE1-NCKAP1 proteins complexed with WT or R87C CYFIP2. Our simulations showed a flexibilization of the loop comprising residues 80–110 due to the loss of contacts between internal residues in the R87C CYFIP2 as well as the key role of residues R/C87, E624, and E689 in structural modification. These data could explain the mechanism by which the mutation impairs the stability and proper regulation of the WRC.     

Early Postoperative Immunothrombosis of Bioprosthetic Mitral Valve and Left Atrium: A Case Report

A 72-year-old female patient with mixed rheumatic mitral valve disease and persistent atrial fibrillation underwent mitral valve replacement and suffered from a combined thrombosis of the bioprosthetic valve and the left atrium as soon as 2 days post operation. The patient immediately underwent repeated valve replacement and left atrial thrombectomy. Yet, four days later the patient died due to the recurrent prosthetic valve and left atrial thrombosis which both resulted in an extremely low cardiac output. In this patient’s case, the thrombosis was notable for the resistance to anticoagulant therapy as well as for aggressive neutrophil infiltration and release of neutrophil extracellular traps (NETs) within the clot, as demonstrated by immunostaining. The reasons behind these phenomena remained unclear, as no signs of sepsis or contamination of the BHV were documented, although the patient was diagnosed with inherited thrombophilia that could impede the fibrinolysis. The described case highlights the hazard of immunothrombosis upon valve replacement and elucidates its mechanisms in this surgical setting.     

Effects of Low-Intensity and Long-Term Aerobic Exercise on the Psoas Muscle of mdx Mice: An Experimental Model of Duchenne Muscular Dystrophy

Duchenne muscular dystrophy (DMD) is a muscle disease characterized by the absence of the protein dystrophin, which causes a loss of sarcolemma integrity, determining recurrent muscle injuries, decrease in muscle function, and progressive degeneration. Currently, there is a need for therapeutic treatments to improve the quality of life of DMD patients. Here, we investigated the effects of a low-intensity aerobic training (37 sessions) on satellite cells, peroxisome proliferator-activated receptor-gamma coactivator (PGC)-1α protein (PGC-1α), and different types of fibers of the psoas muscle from mdx mice (DMD experimental model). Wildtype and mdx mice were randomly divided into sedentary and trained groups (n = 24). Trained animals were subjected to 37 sessions of low-intensity running on a motorized treadmill. Subsequently, the psoas muscle was excised and analyzed by immunofluorescence for dystrophin, satellite cells, myosin heavy chain (MHC), and PGC-1α content. The minimal Feret’s diameters of the fibers were measured, and light microscopy was applied to observe general morphological features of the muscles. The training (37 sessions) improved morphological features in muscles from mdx mice and caused an increase in the number of quiescent/activated satellite cells. It also increased the content of PGC-1α in the mdx group. We concluded that low-intensity aerobic exercise (37 sessions) was able to reverse deleterious changes determined by DMD.     

Two-dimensional linear model of elastic shell accounting for general anisotropy of material

A two-dimensional linearly elastic model of shells made of an anisotropic material described by 21 elastic moduli is developed. For this aim, the generalized Timoshenko–Reissner hypotheses are used. In contrast to the ordinary shell models, the tangential stress-resultants here depend not only on the tangential strains but also on the transverse shear. An asymptotic analysis of the obtained equations is fulfilled. The typical stress-strain states, namely the membrane state, the edge effect state, and the boundary layer, are constructed. The system of the Donnell type is delivered. As an example of general anisotropy, a composite material consisting of a matrix reinforced by the system of fibers inclined to the mid-surface is studied. Certain simple static problems and free vibration problems are solved for a cylindrical shell made of this material.     

Injection of Microporous Annealing Particle (MAP) Hydrogels in the Stroke Cavity Reduces Gliosis and Inflammation and Promotes NPC Migration to the Lesion

With the number of deaths due to stroke decreasing, more individuals are forced to live with crippling disability resulting from the stroke. To date, no therapeutics exist after the first 4.5 h after the stroke onset, aside from rest and physical therapy. Following stroke, a large influx of astrocytes and microglia releasing proinflammatory cytokines leads to dramatic inflammation and glial scar formation, affecting brain tissue's ability to repair itself. Pathological conditions, such as a stroke, trigger neural progenitor cells (NPCs) proliferation and migration toward the damaged site. However, these progenitors are often found far from the cavity or the peri‐infarct tissue. Poststroke tissue remodeling results in a compartmentalized cavity that can directly accept a therapeutic material injection. Here, this paper shows that the injection of a porous hyaluronic acid hydrogel into the stroke cavity significantly reduces the inflammatory response following stroke while increasing peri‐infarct vascularization compared to nonporous hydrogel controls and stroke only controls. In addition, it is shown that the injection of this material impacts NPCs proliferation and migration at the subventricular zone niche and results, for the first time, in NPC migration into the stroke site.     

Take care of your belongings today – securing accessibility to complex electronic business processes

Most of today’s business processes are based solely on digital data. Input, output, and intermediate results are pure digital objects. Keeping such data accessible and meaningful but even more importantly keeping the business processes functional is challenging. With regards to complex electronic business processes traditional archiving does not provide satisfactory results. Due to the fast technical life-cycle the time gap between archiving and re-enactment or reuse of a process poses risks and increases the uncertainty on achievable results. We propose a novel strategy bridging this specific time gap. For this we present a scalable and cost-effective infrastructure with associated workflows focusing on a process’ execution context. Most importantly the process’ developers are able to assess the preserved results in a timely manner, and thus, reduce the uncertainty on future re-enactment results.     

AICAR Protects Vascular Endothelial Cells from Oxidative Injury Induced by the Long-Term Palmitate Excess

Hyperlipidemia manifested by high blood levels of free fatty acids (FFA) and lipoprotein triglycerides is critical for the progression of type 2 diabetes (T2D) and its cardiovascular complications via vascular endothelial dysfunction. However, attempts to assess high FFA effects in endothelial culture often result in early cell apoptosis that poorly recapitulates a much slower pace of vascular deterioration in vivo and does not provide for the longer-term studies of endothelial lipotoxicity in vitro. Here, we report that palmitate (PA), a typical FFA, does not impair, by itself, endothelial barrier and insulin signaling in human umbilical vein endothelial cells (HUVEC), but increases NO release, reactive oxygen species (ROS) generation, and protein labeling by malondialdehyde (MDA) hallmarking oxidative stress and increased lipid peroxidation. This PA-induced stress eventually resulted in the loss of cell viability coincident with loss of insulin signaling. Supplementation with 5-aminoimidazole-4-carboxamide-riboside (AICAR) increased endothelial AMP-activated protein kinase (AMPK) activity, supported insulin signaling, and prevented the PA-induced increases in NO, ROS, and MDA, thus allowing to maintain HUVEC viability and barrier, and providing the means to study the long-term effects of high FFA levels in endothelial cultures. An upgraded cell-based model reproduces FFA-induced insulin resistance by demonstrating decreased NO production by vascular endothelium.