Enterohemorrhagic Escherichia coli (EHEC) outbreaks are commonly associated with contaminated food sources. Unlike normal intestinal bacteria, EHEC are lysogens of lambdoid bacteriophages that also carry a gene for Shiga toxin. Oxidative attack by the immune system or other stressors on the bacterial host can activate the lytic pathway of the latent phage genome to produce phage progeny and the release of Shiga toxin into the surrounding tissues. Within the genomes of bacteriophage λ and Shiga toxin-expressing (Stx+) phages such as φ24B and φP27, there is a conserved set of open reading frames that is located between the exo and xis genes that influences the lysogenic-lytic decision. In this report, we have focused on the largest exo-xis region open reading frame termed ea22 that has been shown previously to have prolysogenic properties. Using a variety of biophysical and bioinformatic methods, we demonstrate that λ and φP27 Ea22 proteins are tetrameric in solution and can be considered in terms of an amino-terminal region, a central coiled-coil region, and a carboxy-terminal region. The carboxy-terminal regions of λ and φ24B Ea22, expressed on their own, form dimers with exceptional thermostability. Limited proteolysis of φP27 Ea22 also identified a C-terminal region along the predicted boundaries. While the three Ea22 proteins all appear to have the hallmarks of a domain in their respective C-terminal regions, each sequence is remarkably dissimilar. To reconcile this difference among Ea22 proteins from λ and Stx+ phages alike, we speculate that each Ea22 may achieve the same function by targeting different components of the same regulatory process in the host.

Considering the large consumption of nicotine and its sedative/stimulant effect on different organs of the body, the detection of low concentration of this material and its subsequent effect on live animals plays a significant role. Optical detection techniques such as plasmonics are the pioneers in highly sensitive detection techniques. However, for investigating the nicotine/smoke effect on live cells, not only the interaction between cell nicotine should be optimized but also the plasmonic interface should show a high sensitivity to the reception of nicotine by the cell receptors. In this study, the sensitivity of the plasmonic detection system was greatly increased using the coupling of plasmon and fluorophore. This coupling could enhance the main plasmonic signal several orders of magnitude besides improving Δ and Ψ ellipsometry parameters. Benefiting from the green fluorescence proteins, the phase shift and the amplitude ratio between the reflections under s- and p-polarized light enhance considerably which verifies the coupling of the dipole of the fluorescence emitter and the plasmons of the metal nanostructure. For 1 s increase of the maintenance time, we encountered a considerable increase in the Δ values that were 0.15° for T e = 1 s and 0.24° for T e = 3 s. Benefiting from extracted ellipsometry parameters, this study could open new avenues toward studying the effect of various types of drugs and stimulants on biological samples using a novel plasmophore platform.

Simultaneous improvement in the mechanical properties and lifetime of polymer nanocomposites is crucially significant to further extend the versatility of polymer materials and reduce environmental impact. In this study, we fabricated reinforced polypropylene (PP)-based nanocomposites with improved aging stability by the addition of surface-modified well-ordered silica nanospheres with a silane coupling agent (SCA) containing hindered phenol antioxidant as a filler. Uniform grafting of the SCA on the filler surface contributed to homogeneous dispersion of the filler into the matrix, leading to improved properties (e.g., stiffness and ductility) and uniform distribution of the antioxidant component into the entire nanocomposite by filler dispersion. The grafting of SCA also likely provides an inhibitory effect on antioxidant migration, which leads to loss of polymer stability during the aging process. This novel idea for the material design of PP-based nanocomposites, which simultaneously enhances their mechanical properties and lifetime, is promising for application in the fabrication of various types of polymer nanocomposites.

Oxidative stress occurs when physiological antioxidant systems do not manage to counteract the excessive intracellular production of reactive oxygen species (ROS), which accumulate leading to irreversible oxidation of DNA and other biomacromolecules, and thus to the onset of pathological conditions. Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is a neurodegenerative disease characterized by autosomal recessive mutations in the sacsin gene (SACS). It has been demonstrated that cells of ARSACS patients show bioenergetic and mitochondrial impairment, denoted by reduced respiratory chain activities and ATP synthesis. In order to design a suitable therapy for ARSACS, it is essential to consider that treatments need to cross the blood-brain barrier (BBB), a specialized structure that separates the subtle environment of the brain from blood circulation. Nanostructured lipid carriers (NLCs), constituted by a solid lipid shell and a liquid lipid phase in the core, have been fabricated for loading hydrophobic molecules, improving their bioavailability. Idebenone (IDE), a synthetic analogue of coenzyme Q10, is able to inhibit lipid peroxidation and detoxify several free radicals. However, because of its poor solubility, it requires ad hoc drug-delivery systems for enhancing its pharmacokinetic properties, preventing undesired cytotoxicity. In this work, NLCs loaded with idebenone (IDE-NLCs) have been prepared. The nanovectors have been physicochemically characterized, and their biological activity has been evaluated on different central nervous system cell lines. IDE-NLCs demonstrated to be stable in water and in cell culture media, and showed a sustained drug release profile. Interestingly, preliminary data demonstrated their ability to permeate an in vitro BBB model. Their protective antioxidant activity in human healthy primary skin fibroblasts and their therapeutic efficacy in ARSACS-derived primary skin fibroblasts have been also investigated, showing their potential for future development as therapeutic agents.

A study of the electrical properties of metallic nanowires requires a clear analysis of conductive networks. In this study, we demonstrated that the conducting networks of Ag nanowires (AgNW) could be visually observed by examination of the voltage contrast of the scanning electron microscopy (SEM) images, which was caused by the differences in the degrees of charging of AgNWs. When AgNWs dispersed on a quartz glass were irradiated by primary electrons, the substrate became negatively charged. This induced positive charges on the AgNWs in contact with the electrodes. As a result, AgNW networks connected to electrodes appeared dark in the SEM image, while the isolated AgNWs appeared brighter. By varying the acceleration voltage of the primary electrons, the extent of charging could be controlled, which, in turn, enabled the observation of the voltage contrast of AgNWs. Using the voltage contrast of SEM images, we could visually distinguish the AgNW networks having an electrical connection with the electrode from the ones that were not connected to the electrode.

The well-known ability to selectively drive nanomagnetic materials coated with anticancer drugs into tumor cells suggested the synthesis and the characterization of magnetic nanoparticles (MNPs) functionalized with (R)-9-acetoxystearic acid, the acetic ester of (R)-9-hydroxystearic acid (9-HSA), an antiproliferative agent active against different cancer cells. The acyl chloride of (R)-9-acetoxystearic acid, synthesized in two steps from 9-HSA, was reacted with (3-aminopropyl)triethoxysilane, chosen as a linker between MNPs and the stearyl moiety. In the last step, the novel amide was bound to magnetite NPs by reaction with silyl groups. A detailed structural, chemical, and magnetic characterization of the obtained material proved that it possesses properties in agreement with the requirements for drug delivery, opening the possibility to further insights focused on the 9-HSA biomedical applications.

Sol-gel-derived bioactive glass nanoparticles have attracted special interest due to their potential as novel therapeutic and regenerative agents. Significant challenges are yet to be addressed. The fabrication of sol-gel-derived nanoparticles in binary and ternary systems with an actual composition that meets the nominal has to be achieved. This work addresses this challenge and delivers nanoparticles in a ternary system with tailored composition and particle size. It also studies how specific steps in the fabrication process can affect the incorporation of the metallic ions, nanoparticle size, and mesoporosity. Sol-gel-derived bioactive glass nanoparticles in the 62 SiO2-34.5 CaO-3.2 P2O5 (mol %) system have been fabricated and characterized for their structural, morphological, and elemental characteristics using Fourier transform infrared spectroscopy, X-ray diffraction analysis, scanning electron microscopy associated with elemental analysis, transmission electron microscopy, and solid-state nuclear magnetic resonance. The fabricated nanoparticles were additionally observed to form the apatite phase when immersed in simulated body fluid. This work highlights the effect of the different processing variables, such as the nature of the solvent, the order in which reagents are added, stirring time, and the concentrations in the catalytic solution on the controlled incorporation of specific ions (e.g., P and Ca) in the nanoparticle network and particle size.

Recurrent infections of neurotropic herpes simplex virus-1 (HSV-1) have been implicated in etiology and pathology of Alzheimer's disease (AD). Although protein and peptide aggregation events are at the center of the AD pathophysiology, except a single study where a peptide derived from glycoprotein B of HSV-1 was reported to form β-amyloid-like aggregates, similar investigations with the entire proteome of HSV-1 have not been attempted. In the current study, 70 HSV-1 proteins were screened using bioinformatics tools to identify aggregation-prone candidates. Thereafter, the 20S proteasome cleavage sites within the sequence of the selected proteins were determined using Pcleavage and NetChop algorithms, thereby mimicking a cellular proteasomal activity providing short peptides. Here, we report the biochemical characterization of a 28-residue-long peptide (HSV-1 gK208-235) derived from glycoprotein K of HSV-1. The peptide showed high aggregation propensity and homology to the C-terminus of Aβ1-42 peptide. The aggregates of gK208-235 peptide were characterized by the Congo red and Thioflavin T assays and Fourier transform infrared (FTIR) spectroscopy, and their spheroid oligomeric structure was established by atomic force microscopy (AFM). Furthermore, the aggregates demonstrated dose-dependent cytotoxicity to primary mouse splenocytes. The current findings hypothesize a mechanism by which HSV-1 may contribute to AD, which may be pursued further in the future.

Efficient and selective transport of rhodium(III) across a polymer inclusion membrane (PIM) from a 0.1 mol dm-3 HCl feed solution, also containing iron(III), to a receiving solution containing 0.1 mol dm-3 HCl and 4.9 mol dm-3 NH4Cl was achieved using a phosphonium-type ionic liquid, trioctyl(dodecyl)phosphonium chloride (P88812Cl), as the metal ion carrier. The optimum PIM composition for the Rh(III) transport was 50 wt % poly(vinylidene-co-hexafluoropropylene) (PVDF-HFP), 30 wt % P88812Cl, and 20 wt % plasticizer 2-nitrophenyl octyl ether (2NPOE). The driving force for the Rh(III) transport was suggested to be the concentration difference of the chloride ion between the feed and the receiving solutions. More than 70% rhodium(III) could be recovered from the receiving solution, and no transport of iron(III) was observed; however, the two metal ions cannot be separated by liquid-liquid extraction. This is the first report of selective transport of rhodium(III) across a polymer inclusion membrane.

We previously showed that commercially available rice peptide Oryza Peptide-P60 (OP60) increased the intracellular glutathione levels. This study aimed to evaluate the antioxidant potential of this peptide and assess its mechanism of action. Pretreatment of HepG2 cells with OP60 reduced the cytotoxicity caused by H2O2 or acetaminophen (APAP) (47.7 ± 1.3% or 12.2 ± 1.3% of the cytotoxicity for 5 mg/mL OP60 pretreatment compared to that in H2O2- or APAP-treated groups, respectively; p < 0.01) through the restoration of glutathione homeostasis. Moreover, OP60 elevated the mRNA level of genes encoding heavy and light subunits of γ-glutamylcysteine synthetase (γ-GCS) by 2.9 ± 0.1-fold and 2.7 ± 0.2-fold (p < 0.001), respectively, at 8 h and also increased the level of mRNA encoding other antioxidant enzymes. Besides, OP60 promoted Nrf2 nuclear translocation by 2.2 ± 0.3-fold (p < 0.05) after 8 h. Conversely, knockdown of Nrf2 inhibited the increase of the intracellular glutathione levels and suppressed the induction of antioxidant enzyme expression by OP60. In animal studies, OP60 prevented APAP-induced liver injury by suppressing glutathione depletion (from 0.19 ± 0.02 mmol/mg protein to 0.90 ± 0.02 mmol/mg protein; p < 0.01, by pretreatment with 500 mg/kg OP60) and increasing heavy subunit of γ-GCS and heme oxygenase-1 expression in the liver. Our results indicated that OP60 exhibits a cytoprotective effect via the Nrf2 signaling pathway and is one of the few peptides with excellent antioxidant properties.

Biohybrid microswimmers, which are realized through the integration of motile microscopic organisms with artificial cargo carriers, have a significant potential to revolutionize autonomous targeted cargo delivery applications in medicine. Nonetheless, there are many open challenges, such as motility performance and immunogenicity of the biological segment of the microswimmers, which should be overcome before their successful transition to the clinic. Here, we present the design and characterization of a biohybrid microswimmer, which is composed of a genetically engineered peritrichously flagellated Escherichia coli species integrated with red blood cell-derived nanoliposomes, also known as nanoerythrosomes. Initially, we demonstrated nanoerythrosome fabrication using the cell extrusion technique and characterization of their size and functional cell membrane proteins with dynamic light scattering and flow cytometry analyses, respectively. Then, we showed the construction of biohybrid microswimmers through the conjugation of streptavidin-modified bacteria with biotin-modified nanoerythrosomes by using non-covalent streptavidin interaction. Finally, we investigated the motility performance of the nanoerythrosome-functionalized biohybrid microswimmers and compared it with the free-swimming bacteria. The microswimmer design approach presented here could lead to the fabrication of personalized biohybrid microswimmers from patients' own cells with high fabrication efficiencies and motility performances.

We conducted a longitudinal study in patients with pancreatic and colorectal cancer. We determined the effect of chemotherapy on extracellular vesicle tissue factor (EVTF) activity and the association of plasma EVTF activity with venous thromboembolism (VTE) and survival.

To compare publicly available rehabilitation protocols designated for rotator cuff (RTC) repairs published online by academic residency programs and private practice institutions.

In line with the theory of event coding, many studies on tool use show that perceived visual and haptic information interacts with action execution. In two experiments, we investigated the temporal persistence of after-effects within an event file, and after-effects in temporally overlapping event files with the n-1 replication task. Each trial consisted of two phases: In phase 1, participants moved a cursor with a pen on a covered tablet while a gain varied the relation between hand and cursor amplitude (Experiment 1). In phase 2, participants replicated the hand amplitude of phase 1 of the previous trial without visual feedback. Any systematic over- and undershoot would be indicative for after-effects. When the cursor amplitude varied and the hand amplitude remained constant, we did not find any after-effects but adjustment of the internal model. For varying hand amplitudes, after-effects appeared in terms of a contrast and assimilation effect between temporally overlapping event files and within an event file, respectively. In Experiment 2, we confirmed that the observed pattern of over- and undershoots fully reflect assimilation/contrast due to perception-action interaction. The findings extend the current view on the temporal stability of short-term binding in sensorimotor transformation tasks: In the n-1 replication task, after-effects appeared only in trials with varying hand amplitudes. We replicated the contrast effect and assimilation effect, and the assimilation effect persisted for up to approximately 20 s.

[123I]epidepride is a high-affinity radiotracer used in single-photon emission computed tomography (SPECT) imaging of the D2/3 receptors. It binds with high affinity to striatal and extrastriatal receptors. Nevertheless, its slow kinetics in the striatum impedes quantification in this region. Thus, an approach that would allow a simultaneous quantification of both striatal and extrastriatal D2/3 receptors would be of interest for preclinical and clinical SPECT neuroimaging. We describe a partial saturation protocol that allows us to produce an in vivo Scatchard plot and thus estimate Bavail and appKd separately in both striatal and extrastriatal regions, through a single dynamic SPECT session. To validate this approach, a multi-injection protocol is used for the full kinetic modeling of [123I]epidepride using a two-tissue compartment, 5-parameter model (2T-5k).

Asiaticoside (AS) has been reported to have protective effect on pulmonary fibrosis (PF). In this study, we aimed to explore the potential mechanism of the therapeutic role of AS and its relationship with A2AR in PF. Adenosine 2A receptor gene knockout (A2AR-/- ) mice and wild-type (WT) mice were used to establish bleomycin (BLM)-induced PF models and were then treated with AS (50 mg/kg/d). Pulmonary inflammation and fibrosis were observed in the PF model with much higher severity in A2AR-/- mice than that in WT mice and AS significantly alleviated lung inflammation and fibrosis; however, it was less effective in A2AR-/- mice than in WT mice via histopathological analysis. Using RNA sequencing analysis, we found up-regulated differentially expressed genes (DEGs) in BLM group were enriched in immune and inflammation-associated pathways compared with control group. There were 242 common DEGs between down-regulated in BLM vs control group and up-regulated in BLM + AS vs BLM group, which were enriched in cAMP and Rap1 signalling pathways. Furthermore, the expression of five key factors of these two pathways including adenylate cyclase (ADCY1, ADCY5, ADCY8, cAMP and Rap1) were confirmed up-regulated by AS with the presence of A2AR. Therefore, AS might attenuate BLM-induced PF by activating cAMP and Rap1 signalling pathways which is assisted by A2AR, making it a promising therapeutic optional for PF.

Ever reports showed that PCNP is associated with human cancers including neuroblastoma and lung cancer. However, the role and underlying molecular mechanism of PCNP in ovarian cancer have not been plenty elucidated. Herein, we first investigated the expression of PCNP in ovarian cancer tissues and cells, the effects of PCNP in ovarian cancer proliferation, apoptosis, migration and invasion, and determined the molecular mechanism of PCNP in ovarian cancer progression. The results indicated that PCNP was significantly overexpressed in human ovarian cancer tissues and cells, and related to poor prognosis in ovarian cancer patients. In addition, we also detected that PCNP promoted ovarian cancer cells growth, migration and invasion, as well as inhibited ovarian cancer cells apoptosis. Mechanistically, PCNP binding to β-catenin promoted β-catenin nuclear translocation and further activated Wnt/β-catenin signalling pathway. Moreover, PCNP regulated the expression of genes involved in EMT and further triggered EMT occurrence. Conclusionally, PCNP may promote ovarian cancer progression through activating Wnt/β-catenin signalling pathway and EMT, acting as a novel and promising target for treating ovarian cancer.

This study assessed the safety and efficacy of three different doses of BoNT-A for persistent myofascial pain (MFP). One hundred female subjects were randomly assigned into five groups (n = 20): oral appliance (OA), saline solution (SS) and three BoNT-A groups with different doses. Pain intensity and pressure pain threshold were evaluated up to 24 weeks after treatment. Adverse effects related to muscle contraction, masticatory performance, muscle thickness and mandibular bone volume were also assessed. Changes over time were compared within and between groups. The "nparLD" package and Wilcoxon signed-rank test were used to analyze the data. BoNT-A reduced pain intensity (p < 0.0001) and increased pressure pain threshold (p < 0.0001) for up to 24 weeks compared to the placebo. No differences were found between BoNT-A and OA at the last follow-up. A transient decline in masticatory performance (p < 0.05) and muscle contraction (p < 0.0001), and a decrease in muscle thickness (p < 0.05) and coronoid and condylar process bone volume (p < 0.05) were found as dose-related adverse effects of BoNT-A. Regardless of the dose, BoNT-A was as effective as OA on MFP. Notwithstanding, due to BoNT-A dose-related adverse effects, we suggest the use of low doses of BoNT-A in MFP patients that do not benefit from conservative treatments.

As more cases of COVID-19 are studied and treated worldwide, it had become apparent that the lethal and most severe cases of pneumonia are due to an out-of-control inflammatory response to the SARS-CoV-2 virus. I explored the putative causes of this specific feature through a detailed genomic comparison with the closest SARS-CoV-2 relatives isolated from bats, as well as previous coronavirus strains responsible for the previous epidemics (SARS-CoV and MERS-CoV). The high variability region of the nsp3 protein was confirmed to exhibit the most variations between closest strains. It was then studied in the context of physiological and molecular data available in the literature. A number of convergent findings suggest de-mono-ADP-ribosylation (de-MARylation) of STAT1 by the SARS-CoV-2 nsp3 as a putative cause of the cytokine storm observed in the most severe cases of COVID-19. This may suggest new therapeutic approaches and help in designing assays to predict the virulence of naturally circulating SARS-like animal coronaviruses.

We aimed to evaluate sleep and sleep-related physiological parameters (heart rate variability and glucose dynamics) among evacuees by experimentally recreating the sleep environment of evacuation shelters and cars. Nine healthy young male subjects participated in this study. Two interventions, modeling the sleep environments of evacuation shelters (evacuation shelter trial) and car seats (car trial), were compared with sleep at home (control trial). Physiological data were measured using portable two-channel electroencephalogram and electrooculogram monitoring systems, wearable heart rate sensors, and flash glucose monitors. Wake after sleep onset (WASO) and stage shift were greater in both intervention trials than the control trial, while rapid-eye movement (REM) latency and non-rapid eye movement (NREM) 1 were longer and REM duration was shorter in the evacuation shelter trial than the control trial. Glucose dynamics and power at low frequency (LF.p) of heart rate variability were higher in the car trial than in the control trial. It was confirmed that sleep environment was important to maintain sleep, and affected glucose dynamics and heart rate variability in the experimental situation.