Repurposing of neprilysin inhibitor 'sacubitrilat' as an anti-cancer drug by modulating epigenetic and apoptotic regulators.

Modifications in the epigenetic landscape have been considered a hallmark of cancer. Histone deacetylation is one of the crucial epigenetic modulations associated with the aggressive progression of various cancer subtypes. Herein, we have repurposed the neprilysin inhibitor sacubitrilat as a potent anticancer agent using in-silico protein-ligand interaction profiler (PLIP) analysis, molecular docking, and in vitro studies. The screening of PLIP profiles between vorinostat/panobinostat and HDACs/LTA4H followed by molecular docking resulted in five (Sacubitrilat, B65, BDS, BIR, and NPV) FDA-approved, experimental and investigational drugs. Sacubitrilat has demonstrated promising anticancer activity against colorectal cancer (SW-480) and triple-negative breast cancer (MDA-MB-231) cells, with IC50 values of 14.07 µg/mL and 23.02 µg/mL, respectively. FACS analysis revealed that sacubitrilat arrests the cell cycle at the G0/G1 phase and induces apoptotic-mediated cell death in SW-480 cells. In addition, sacubitrilat inhibited HDAC isoforms at the transcriptomic level by 0.7-0.9 fold and at the proteomic level by 0.5-0.6 fold as compared to the control. Sacubitrilat increased the protein expression of tumor-suppressor (p53) and pro-apoptotic makers (Bax and Bid) by 0.2-2.5 fold while decreasing the expression of anti-apoptotic Bcl2 and Nrf2 proteins by 0.2-0.5 fold with respect to control. The observed cleaved PARP product indicates that sacubitrilat induces apoptotic-mediated cell death. This study may pave the way to identify the anticancer potential of sacubitrilat and can be explored in human clinical trials.

Electrostatic interactions between ions near Thomas-Fermi substrates and the surface energy of ionic crystals at imperfect metals.

The electrostatic interaction between two charged particles is strongly modified in the vicinity of a metal. This situation is usually accounted for by the celebrated image charges approach, which was further extended to account for the electronic screening properties of the metal at the level of the Thomas-Fermi description. In this paper we build upon a previous approach [M. A. Vorotyntsev and A. A. Kornyshev, Zh. Eksp. Teor. Fiz., 1980, 78(3), 1008-1019] and successive works to calculate the 1-body and 2-body electrostatic energy of ions near a metal in terms of the Thomas-Fermi screening length. We propose workable approximations suitable for molecular simulations of ionic systems close to metallic walls. Furthermore, we use this framework to calculate analytically the electrostatic contribution to the surface energy of a one dimensional crystal at a metallic wall and its dependence on the Thomas-Fermi screening length. These calculations provide a simple interpretation for the surface energy in terms of image charges, which allows for an estimation of the interfacial properties in more complex situations of a disordered ionic liquid close to a metal surface. The counter-intuitive outcome is that electronic screening, as characterized by a molecular Thomas-Fermi length lTF, profoundly affects the wetting of ionic systems close to a metal, in line with the recent experimental observation of capillary freezing of ionic liquids in metallic confinement.

Influence of border disease virus (BDV) on serological surveillance within the bovine virus diarrhea (BVD) eradication program in Switzerland.

BACKGROUND: In 2008, a program to eradicate bovine virus diarrhea (BVD) in cattle in Switzerland was initiated. After targeted elimination of persistently infected animals that represent the main virus reservoir, the absence of BVD is surveilled serologically since 2012. In view of steadily decreasing pestivirus seroprevalence in the cattle population, the susceptibility for (re-) infection by border disease (BD) virus mainly from small ruminants increases. Due to serological cross-reactivity of pestiviruses, serological surveillance of BVD by ELISA does not distinguish between BVD and BD virus as source of infection. RESULTS: In this work the cross-serum neutralisation test (SNT) procedure was adapted to the epidemiological situation in Switzerland by the use of three pestiviruses, i.e., strains representing the subgenotype BVDV-1a, BVDV-1h and BDSwiss-a, for adequate differentiation between BVDV and BDV. Thereby the BDV-seroprevalence in seropositive cattle in Switzerland was determined for the first time. Out of 1,555 seropositive blood samples taken from cattle in the frame of the surveillance program, a total of 104 samples (6.7%) reacted with significantly higher titers against BDV than BVDV. These samples originated from 65 farms and encompassed 15 different cantons with the highest BDV-seroprevalence found in Central Switzerland. On the base of epidemiological information collected by questionnaire in case- and control farms, common housing of cattle and sheep was identified as the most significant risk factor for BDV infection in cattle by logistic regression. CONCLUSION: This indicates that pestiviruses from sheep should be considered as a source of infection of domestic cattle and might well impede serological BVD surveillance.

ac Wien Effect in Spin Ice, Manifest in Nonlinear, Nonequilibrium Susceptibility.

The Wien effect is a model process for field-induced charge creation. Here it is derived for a nonelectrical system: the spin ice "magnetolyte"-a unique system showing perfect charge symmetry. An entropic reaction field, analogous to the Jaccard field in ice, opposes direct current, but a frequency window exists in which the Wien effect for magnetolyte and electrolyte are indistinguishable. The universal enhancement of monopole density speeds up the magnetization dynamics, which manifests in the nonlinear, nonequilibrium ac susceptibility. This is a rare instance where such effects may be calculated, providing new insights for electrolytes. Experimental predictions are made for Dy2Ti2O7 spin ice.

Motor imagery-induced EEG patterns in individuals with spinal cord injury and their impact on brain-computer interface accuracy.

OBJECTIVE: Assimilating the diagnosis complete spinal cord injury (SCI) takes time and is not easy, as patients know that there is no 'cure' at the present time. Brain-computer interfaces (BCIs) can facilitate daily living. However, inter-subject variability demands measurements with potential user groups and an understanding of how they differ to healthy users BCIs are more commonly tested with. Thus, a three-class motor imagery (MI) screening (left hand, right hand, feet) was performed with a group of 10 able-bodied and 16 complete spinal-cord-injured people (paraplegics, tetraplegics) with the objective of determining what differences were present between the user groups and how they would impact upon the ability of these user groups to interact with a BCI. APPROACH: Electrophysiological differences between patient groups and healthy users are measured in terms of sensorimotor rhythm deflections from baseline during MI, electroencephalogram microstate scalp maps and strengths of inter-channel phase synchronization. Additionally, using a common spatial pattern algorithm and a linear discriminant analysis classifier, the classification accuracy was calculated and compared between groups. MAIN RESULTS: It is seen that both patient groups (tetraplegic and paraplegic) have some significant differences in event-related desynchronization strengths, exhibit significant increases in synchronization and reach significantly lower accuracies (mean (M) = 66.1%) than the group of healthy subjects (M = 85.1%). SIGNIFICANCE: The results demonstrate significant differences in electrophysiological correlates of motor control between healthy individuals and those individuals who stand to benefit most from BCI technology (individuals with SCI). They highlight the difficulty in directly translating results from healthy subjects to participants with SCI and the challenges that, therefore, arise in providing BCIs to such individuals.

Onsager's Wien effect on a lattice.

The second Wien effect describes the nonlinear, non-equilibrium response of a weak electrolyte in moderate to high electric fields. Onsager's 1934 electrodiffusion theory, along with various extensions, has been invoked for systems and phenomena as diverse as solar cells, surfactant solutions, water splitting reactions, dielectric liquids, electrohydrodynamic flow, water and ice physics, electrical double layers, non-ohmic conduction in semiconductors and oxide glasses, biochemical nerve response and magnetic monopoles in spin ice. In view of this technological importance and the experimental ubiquity of such phenomena, it is surprising that Onsager's Wien effect has never been studied by numerical simulation. Here we present simulations of a lattice Coulomb gas, treating the widely applicable case of a double equilibrium for free charge generation. We obtain detailed characterization of the Wien effect and confirm the accuracy of the analytical theories as regards the field evolution of the free charge density and correlations. We also demonstrate that simulations can uncover further corrections, such as how the field-dependent conductivity may be influenced by details of microscopic dynamics. We conclude that lattice simulation offers a powerful means by which to model and investigate system-specific corrections to the Onsager theory, and thus constitutes a valuable tool for detailed theoretical studies of the numerous practical applications of the second Wien effect.

Watching object related movements modulates mirror-like activity in parietal brain regions.

OBJECTIVE: We studied the activation of cortical motor and parietal areas during the observation of object related grasping movements. By manipulating the type of an object (realistic versus abstract) and the type of grasping (correct versus incorrect), we addressed the question how observing such object related movements influences cortical rhythmicity, especially the mu-rhythm, in the context of an "extended" human mirror neuron system (MNS). METHODS: Multichannel electroencephalogram (EEG) was recorded during the observation of different object-related grasping actions in twenty healthy subjects. Different movies were presented, showing sequences of correct or incorrect hand grasping actions related to an abstract or realistic (daily life) object. RESULTS: Event-related de/synchronization (ERD/ERS) analyses revealed a larger ERD in the upper alpha (10-12 Hz), beta (16-20 Hz) and gamma (36-40 Hz) frequency bands over parietal brain regions depending on the type of grasping. The type of object only influenced ERD patterns in the gamma band range (36-40 Hz) at parietal sites suggesting a strong relation of gamma band activity and cortical object representation. Abstract and realistic objects produced lower beta band synchronization at central sites only, whereas depending on the type of grasping an ERS in the upper alpha band (10-12 Hz) was observed. CONCLUSION: Depending on the type of the grasped object and the type of grasping stronger parietal cortical activation occurred during movement observation. SIGNIFICANCE: Discussing the results in terms of an "extended" human mirror neuron system (MNS), it could be concluded that beside sensorimotor areas a stronger involvement of parietal brain regions was found depending on the type of object and grasping movement observed.

A new plant sex-linked gene with high sequence diversity and possible introgression of the X copy.

We describe patterns of DNA sequence diversity in a newly identified sex-linked gene, SlX9/SlY9, in Silene latifolia (Caryophyllaceae). The copies on both sex chromosomes seem to be functional, and each maps close to the respective X- and Y-linked copy of another sex-linked gene pair, SlCypX/SlCypY. The Y-linked copy has low diversity, similar to what has been found for several other Y-linked genes in S. latifolia, and consistent with the theoretical expectations of hitch-hiking processes occurring on a non-recombining chromosome. However, SlX9 has higher diversity than other genes on the S. latifolia X chromosome. We evaluate the hypothesis of introgression from the closely related species S. dioica as an explanation for the high sequence diversity observed.

Toward a hybrid brain-computer interface based on imagined movement and visual attention.

Brain-computer interface (BCI) systems do not work for all users. This article introduces a novel combination of tasks that could inspire BCI systems that are more accurate than conventional BCIs, especially for users who cannot attain accuracy adequate for effective communication. Subjects performed tasks typically used in two BCI approaches, namely event-related desynchronization (ERD) and steady state visual evoked potential (SSVEP), both individually and in a 'hybrid' condition that combines both tasks. Electroencephalographic (EEG) data were recorded across three conditions. Subjects imagined moving the left or right hand (ERD), focused on one of the two oscillating visual stimuli (SSVEP), and then simultaneously performed both tasks. Accuracy and subjective measures were assessed. Offline analyses suggested that half of the subjects did not produce brain patterns that could be accurately discriminated in response to at least one of the two tasks. If these subjects produced comparable EEG patterns when trying to use a BCI, these subjects would not be able to communicate effectively because the BCI would make too many errors. Results also showed that switching to a different task used in BCIs could improve accuracy in some of these users. Switching to a hybrid approach eliminated this problem completely, and subjects generally did not consider the hybrid condition more difficult. Results validate this hybrid approach and suggest that subjects who cannot use a BCI should consider switching to a different BCI approach, especially a hybrid BCI. Subjects proficient with both approaches might combine them to increase information throughput by improving accuracy, reducing selection time, and/or increasing the number of possible commands.

Genetic recombination and molecular evolution.

Reduced rates of genetic recombination are often associated with reduced genetic variability and levels of adaptation. Several different evolutionary processes, collectively known as Hill-Robertson (HR) effects, have been proposed as causes of these correlates of recombination. Here, we use DNA sequence polymorphism and divergence data from the noncrossing over dot chromosome of Drosophila to discriminate between two of the major forms of HR effects: selective sweeps and background selection. This chromosome shows reduced levels of silent variability and reduced effectiveness of selection. We show that neither model fits the data on variability. We propose that, in large genomic regions with restricted recombination, HR effects among nonsynonymous mutations undermine the effective strength of selection, so that their background selection effects are weakened. This modified model fits the data on variability and also explains why variability in very large nonrecombining genomes is not completely wiped out. We also show that HR effects of this type can produce an individual selection advantage to recombination, as well as greatly reduce the mean fitness of nonrecombining genomes and genomic regions.

Out of the Andes: patterns of diversification in clearwing butterflies.

Global biodiversity peaks in the tropical forests of the Andes, a striking geological feature that has likely been instrumental in generating biodiversity by providing opportunities for both vicariant and ecological speciation. However, the role of these mountains in the diversification of insects, which dominate biodiversity, has been poorly explored using phylogenetic methods. Here we study the role of the Andes in the evolution of a diverse Neotropical insect group, the clearwing butterflies. We used dated species-level phylogenies to investigate the time course of speciation and to infer ancestral elevation ranges for two diverse genera. We show that both genera likely originated at middle elevations in the Andes in the Middle Miocene, contrasting with most published results in vertebrates that point to a lowland origin. Although we detected a signature of vicariance caused by the uplift of the Andes at the Miocene-Pliocene boundary, most sister species were parapatric without any obvious vicariant barrier. Combined with an overall decelerating speciation rate, these results suggest an important role for ecological speciation and adaptive radiation, rather than simple vicariance.

Expression of mammalian defensin genes.

Antimicrobial peptides are a prevalent mechanism of host defense found throughout nature. In mammals, defensins are among the most abundant of these broad-spectrum antibiotics, and are expressed in epithelial and hematopoietic cells. The defensin peptides are especially abundant in neutrophils; however, gene expression is limited to the promyelocyte stage. In epithelial cells, defensin genes are found as both constitutively expressed and inducible. Induction has been observed in vitro by stimulation with bacterial lipopolysaccharide as well as inflammatory mediators. In vivo, up-regulation of several defensin genes occurs in both infectious and inflammatory states. Gene regulation occurs via signal transduction pathways common to other innate immune responses, utilizing transcription factors such as nuclear factor (NF)-kappaB and NF interleukin-6. Together, the data suggest a broad-based innate host defense whereby potent antimicrobial peptides are present to prevent initial colonization by pathogenic microorganisms. In addition, the recognition of bacteria coupled with a nascent inflammatory response can bolster this defense by a coordinated up-regulation of the peptides.

Transcriptional regulation of beta-defensin gene expression in tracheal epithelial cells.

Innate immunity provides an ever-present or rapidly inducible initial defense against microbial infection. Among the effector molecules of this defense in many species are broad-spectrum antimicrobial peptides. Tracheal antimicrobial peptide (TAP) was the first discovered member of the beta-defensin family of mammalian antimicrobial peptides. TAP is expressed in the ciliated epithelium of the bovine trachea, and its mRNA levels are dramatically increased upon stimulation with bacteria or bacterial lipopolysaccharide (LPS). We report here that this induction by LPS is regulated at the level of transcription. Furthermore, the transfection of reporter gene constructs into tracheal epithelial cells indicates that DNA sequences in the 5' flanking region of the TAP gene, within 324 nucleotides of the transcription start site, are responsible in part for mediating gene induction. This region includes consensus binding sites for NF-kappaB and nuclear factor interleukin-6 (NF IL-6) transcription factors. Gel mobility shift assays indicate that LPS induces NF-kappaB binding activity in the nuclei of these cells, while NF IL-6 binding activity is constitutively present. The gene encoding human beta-defensin 2, a human homologue of TAP with similar inducible expression patterns in the airway, was cloned and found to have conserved NF-kappaB and NF IL-6 consensus binding sites in its 5' flanking region. Previous studies of antimicrobial peptides from insects indicated that their induction by infectious microbes and microbial products also occurs via activation of NF-kappaB-like and NF IL-6-like transcription factors. Together, these observations indicate that a strategy for the induction of peptide-based antimicrobial innate immunity is conserved among evolutionarily diverse organisms.

The influence of experience on the reproducibility of the ankle-brachial systolic pressure ratio in peripheral arterial occlusive disease.

OBJECTIVES: to estimate the intra-observer variability of the measurement of the ankle-brachial systolic pressure index (ABPI) and to compare the reproducibility of the measurements by experienced vascular laboratory assistants and by less-experienced general practice personnel. DESIGN: repeated measurement of ABPI by general practitioners (GPs), GP-assistants and vascular laboratory assistants using a pocket Doppler device and a random-zero sphygmomanometer. METHODS AND MATERIALS: ABPI was measured in six patients with various degrees of PAOD by two experienced observers (vascular laboratory assistants) and by 24 less-experienced observers (18 practice assistants, six GPs). RESULTS: the total number of measurements was 354. The overall intra-observer variability estimate was 11.8% ABPI. The intra-observer variability was 7.3% in the experienced observers and 12.0% in the less-experienced observers. The difference of variability between experienced and less-experienced observers was significant. CONCLUSIONS: the ABPI is suitable in follow-up studies where repeated measurements are needed. Differences between measurements can be minimised by performing repeated measurements or by using more experienced observers.

Risk factors and cardiovascular diseases associated with asymptomatic peripheral arterial occlusive disease. The Limburg PAOD Study. Peripheral Arterial Occlusive Disease.

OBJECTIVES: To describe the risk-factor profile and cardiovascular comorbidity of asymptomatic peripheral arterial occlusive disease (PAOD). DESIGN: A cross-sectional survey. Asymptomatic PAOD was defined as an ankle-brachial pressure index < 0.95, measured on two consecutive occasions, without intermittent claudication. Logistic regression analyses were performed to investigate independent associations between age, gender, smoking status, hypertension, obesity, diabetes, hypercholesterolaemia, physical activity, a family history of cardiovascular disease, the occurrence of ischaemic heart disease and cerebrovascular disease (CeVD) and asymptomatic PAOD. SETTING: 18 general practices in the province of Limburg, the Netherlands. SUBJECTS: A total of 3650 subjects, aged 40-78 years. MAIN RESULTS: Asymptomatic PAOD was present in 8.6% (n = 314) and symptomatic disease in 3.8% (n = 138) of the participants. Age, smoking status, hypertension, and diabetes were significantly associated with asymptomatic PAOD. The ratio of asymptomatic to symptomatic PAOD was higher among the younger age groups. Male gender, hypertension and smoking status were stronger associated with symptomatic PAOD compared with asymptomatic PAOD. Asymptomatic subjects had more IHD and CeVD comorbidity compared with the healthy population. CONCLUSION: Our findings suggest that the risk-factor profile and cardiovascular comorbidity of asymptomatic subjects is comparable to claudicants. Preventive efforts could be made to diminish the influence of especially smoking, diabetes and hypertension in asymptomatic subjects.