An investigation of herpes simplex virus promoter activity compatible with latency establishment reveals VP16-independent activation of immediate-early promoters in sensory neurones

Herpes simplex virus (HSV) type-1 establishes lifelong latency in sensory neurones and it is widely assumed that latency is the consequence of a failure to initiate virus immediate-early (IE) gene expression. However, using a Ore reporter mouse system in conjunction with Ore-expressing HSV-1 recombinants we have previously shown that activation of the IE ICPO promoter can precede latency establishment in at least 30 % of latently infected cells. During productive infection of non-neuronal cells, IE promoter activation is largely dependent on the transactivator VP16 a late structural component of the virion. Of significance, VP16 has recently been shown to exhibit altered regulation in neurones; where its de novo synthesis is necessary for IE gene expression during both lytic infection and reactivation from latency. In the current study, we utilized the Ore reporter mouse model system to characterize the full extent of viral promoter activity compatible with cell survival and latency establishment. In contrast to the high frequency activation of representative IE promoters prior to latency establishment, cell marking using a virus recombinant expressing Ore under VP16 promoter control was very inefficient. Furthermore, infection of neuronal cultures with VP16 mutants reveals a strong VP16 requirement for IE promoter activity in non-neuronal cells, but not sensory neurones. We conclude that only IE promoter activation can efficiently precede latency establishment and that this activation is likely to occur through a VP16-independent mechanism

Oropouche Virus Infection And Pathogenesis Are Restricted By Mavs, Irf-3, Irf-7, And Type I Interferon Signaling Pathways In Nonmyeloid Cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Oropouche virus (OROV) is a member of the Orthobunyavirus genus in the Bunyaviridae family and a prominent cause of insect-transmitted viral disease in Central and South America. Despite its clinical relevance, little is known about OROV pathogenesis. To define the host defense pathways that control OROV infection and disease, we evaluated OROV pathogenesis and immune responses in primary cells and mice that were deficient in the RIG-I-like receptor signaling pathway (MDA5, RIG-I, or MAVS), downstream regulatory transcription factors (IRF-3 or IRF-7), beta interferon (IFN-beta), or the receptor for type I IFN signaling (IFNAR). OROV replicated to higher levels in primary fibroblasts and dendritic cells lacking MAVS signaling, the transcription factors IRF-3 and IRF-7, or IFNAR than in wild-type (WT) cells. In mice, deletion of IFNAR, MAVS, or IRF-3 and IRF-7 resulted in uncontrolled OROV replication, hypercytokinemia, extensive liver damage, and death, whereas WT congenic animals failed to develop disease. Unexpectedly, mice with a selective deletion of IFNAR on myeloid cells (CD11c Cre(+) Ifnar(f/f) or LysM Cre(+) Ifnar(f/f)) did not sustain enhanced disease with OROV or a selective (flox/flox) deletion La Crosse virus, a closely related encephalitic orthobunyavirus. In bone marrow chimera studies, recipient irradiated Ifnar(-/-) mice reconstituted with WT hematopoietic cells sustained high levels of OROV replication and liver damage, whereas WT mice reconstituted with Ifnar(-/-) bone marrow were resistant to disease. Collectively, these results establish a dominant protective role for MAVS, IRF-3 and IRF-7, and IFNAR in restricting OROV infection and tissue injury and suggest that IFN signaling in nonmyeloid cells contributes to the host defense against orthobunyaviruses.89947204737National Institutes of Health [R01 AI104972, P30 DK52574]Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)University Research Committee grantConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CNPq [246513/2012-8

Fat gain with physical detraining is correlated with increased glucose transport and oxidation in periepididymal white adipose tissue in rats

As it is a common observation that obesity tends to occur after discontinuation of exercise, we investigated how white adipocytes isolated from the periepididymal fat of animals with interrupted physical training transport and oxidize glucose, and whether these adaptations support the weight regain seen after 4 weeks of physical detraining. Male Wistar rats (45 days old, weighing 200 g) were divided into two groups (n=10): group D (detrained), trained for 8 weeks and detrained for 4 weeks; and group S (sedentary). The physical exercise was carried out on a treadmill for 60 min/day, 5 days/week for 8 weeks, at 50-60% of the maximum running capacity. After the training protocol, adipocytes isolated from the periepididymal adipose tissue were submitted to glucose uptake and oxidation tests. Adipocytes from detrained animals increased their glucose uptake capacity by 18.5% compared with those from sedentary animals (P<0.05). The same cells also showed a greater glucose oxidation capacity in response to insulin stimulation (34.55%) compared with those from the S group (P<0.05). We hypothesize that, owing to the more intense glucose entrance into adipose cells from detrained rats, more substrate became available for triacylglycerol synthesis. Furthermore, this increased glucose oxidation rate allowed an increase in energy supply for triacylglycerol synthesis. Thus, physical detraining might play a role as a possible obesogenic factor for increasing glucose uptake and oxidation by adipocytes487650653CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação2006/60403-

Multiple voice disorders in the same individual: Investigating handcrafted features, multi-label classification algorithms, and base-learners

Non-invasive acoustic analyses of voice disorders have been at the forefront of current biomedical research. Usual strategies, essentially based on machine learning (ML) algorithms, commonly classify a subject as being either healthy or pathologically-affected. Nevertheless, the latter state is not always a result of a sole laryngeal issue, i.e., multiple disorders might exist, demanding multi-label classification procedures for effective diagnoses. Consequently, the objective of this paper is to investigate the application of five multi-label classification methods based on problem transformation to play the role of base-learners, i.e., Label Powerset, Binary Relevance, Nested Stacking, Classifier Chains, and Dependent Binary Relevance with Random Forest (RF) and Support Vector Machine (SVM), in addition to a Deep Neural Network (DNN) from an algorithm adaptation method, to detect multiple voice disorders, i.e., Dysphonia, Laryngitis, Reinke's Edema, Vox Senilis, and Central Laryngeal Motion Disorder. Receiving as input three handcrafted features, i.e., signal energy (SE), zero-crossing rates (ZCRs), and signal entropy (SH), which allow for interpretable descriptors in terms of speech analysis, production, and perception, we observed that the DNN-based approach powered with SE-based feature vectors presented the best values of F1-score among the tested methods, i.e., 0.943, as the averaged value from all the balancing scenarios, under Saarbrücken Voice Database (SVD) and considering 20% of balancing rate with Synthetic Minority Over-sampling Technique (SMOTE). Finally, our findings of most false negatives for laryngitis may explain the reason why its detection is a serious issue in speech technology. The results we report provide an original contribution, allowing for the consistent detection of multiple speech pathologies and advancing the state-of-the-art in the field of handcrafted acoustic-based non-invasive diagnosis of voice disorders

Interferon-regulatory Factor 5-dependent Signaling Restricts Orthobunyavirus Dissemination To The Central Nervous System

Interferon (IFN)-regulatory factor 5 (IRF-5) is a transcription factor that induces inflammatory responses after engagement and signaling by pattern recognition receptors. To define the role of IRF-5 during bunyavirus infection, we evaluated Oropouche virus (OROV) and La Crosse virus (LACV) pathogenesis and immune responses in primary cells and in mice with gene deletions in Irf3, Irf5, and Irf7 or in Irf5 alone. Deletion of Irf3, Irf5, and Irf7 together resulted in uncontrolled viral replication in the liver and spleen, hypercytokinemia, extensive liver injury, and an early-death phenotype. Remarkably, deletion of Irf5 alone resulted in meningoencephalitis and death on a more protracted timeline, 1 to 2 weeks after initial OROV or LACV infection. The clinical signs in OROV-infected Irf5(-/-) mice were associated with abundant viral antigen and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells in several regions of the brain. Circulating dendritic cell (DC) subsets in Irf5(-/-) mice had higher levels of OROV RNA in vivo yet produced lower levels of type I IFN than wild-type (WT) cells. This result was supported by data obtained in vitro, since a deficiency of IRF-5 resulted in enhanced OROV infection and diminished type I IFN production in bone marrow-derived DCs. Collectively, these results indicate a key role for IRF-5 in modulating the host antiviral response in peripheral organs that controls bunyavirus neuroinvasion in mice.90118920

Predictive biometrics: A review and analysis of predicting personal characteristics from biometric data

Interest in the exploitation of soft biometrics information has continued to develop over the last decade or so. In comparison with traditional biometrics, which focuses principally on person identification, the idea of soft biometrics processing is to study the utilisation of more general information regarding a system user, which is not necessarily unique. There are increasing indications that this type of data will have great value in providing complementary information for user authentication. However, the authors have also seen a growing interest in broadening the predictive capabilities of biometric data, encompassing both easily definable characteristics such as subject age and, most recently, `higher level' characteristics such as emotional or mental states. This study will present a selective review of the predictive capabilities, in the widest sense, of biometric data processing, providing an analysis of the key issues still adequately to be addressed if this concept of predictive biometrics is to be fully exploited in the future

Observation of a topologically protected state in a magnetic domain wall stabilized by a ferromagnetic chemical barrier

The precise control and stabilization of magnetic domain walls is key for the development of the next generation magnetic nano-devices. Among the multitude of magnetic configurations of a magnetic domain wall, topologically protected states are of particular interest due to their intrinsic stability. In this work, using XMCD-PEEM, we have observed a topologically protected magnetic domain wall in a ferromagnetic cylindrical nanowire. Its structure is stabilized by periodic sharp alterations of the chemical composition in the nanowire. The large stability of this topologically protected domain wall contrasts with the mobility of other non-protected and non-chiral states also present in the same nanowire. The micromagnetic simulations show the structure and the conditions required to find the topologically protected state. These results are relevant for the design of future spintronic devices such as domain wall based RF oscillators or magnetic memories

Increasing Acid Concentration, Time and Using a Two-Part Silane Potentiates Bond Strength of Lithium Disilicate-Reinforced Glass Ceramic to Resin Composite:An Exploratory Laboratory Study

There is still a lack of consensus concerning the recommended etching concentration, application time and type of silane when bonding lithium disilicate-reinforced glass ceramics manufactured by CAD/CAM. The purpose of this study was thus to conduct an in vitro study which investigates the influence of hydrofluoric acid (HF) concentration, etching time and silane type on the microtensile bond strength (μTBS) of lithium disilicate to resin composites. Thirty-nine IPS e.max CAD blocks were randomly divided between thirteen groups (n = 3). The variables were HF concentration (9.5 or 4.9%), etching time (20 or 60 s) and silane type (Bis-Silane, Monobond Plus and ESPE Sil Silane). The blocks were cut into beams, aged for 10,000 cycles in a thermocycler and submitted to tensile stress to determine μTBS. A control group featuring the Monobond Etch & Prime (MEP) agent that combines etching/silanisation into a simultaneous process was also added. This group was discarded from the analysis due to only having pre-test failures. The data were analysed using a three-way ANOVA (α = 0.05). The HF concentration, etching time and silane type significantly influenced μTBS (p < 0.001). Significant interactions between time and silane type (p = 0.004), HF concentration and silane type (p < 0.001) and among the three factors (p < 0.001) were noted. Etching lithium disilicate with 9.5% HF (60 s), followed by the application of Bis-Silane, resulted in the highest μTBS (16.6 ± 9.0 MPa). The highest concentration and etching time under study, combined with a two-part silane, resulted in the highest bond strength, while the application of MEP showed a complete pre-test failure

IRF-5-dependent signaling restricts Orthobunyavirus dissemination to the central nervous system

ABSTRACT Interferon (IFN)-regulatory factor 5 (IRF-5) is a transcription factor that induces inflammatory responses after engagement and signaling by pattern recognition receptors. To define the role of IRF-5 during bunyavirus infection, we evaluated Oropouche virus (OROV) and La Crosse virus (LACV) pathogenesis and immune responses in primary cells and in mice with gene deletions in Irf3 , Irf5 , and Irf7 or in Irf5 alone. Deletion of Irf3 , Irf5 , and Irf7 together resulted in uncontrolled viral replication in the liver and spleen, hypercytokinemia, extensive liver injury, and an early-death phenotype. Remarkably, deletion of Irf5 alone resulted in meningoencephalitis and death on a more protracted timeline, 1 to 2 weeks after initial OROV or LACV infection. The clinical signs in OROV-infected Irf5 −/− mice were associated with abundant viral antigen and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells in several regions of the brain. Circulating dendritic cell (DC) subsets in Irf5 −/− mice had higher levels of OROV RNA in vivo yet produced lower levels of type I IFN than wild-type (WT) cells. This result was supported by data obtained in vitro , since a deficiency of IRF-5 resulted in enhanced OROV infection and diminished type I IFN production in bone marrow-derived DCs. Collectively, these results indicate a key role for IRF-5 in modulating the host antiviral response in peripheral organs that controls bunyavirus neuroinvasion in mice. IMPORTANCE Oropouche virus (OROV) and La Crosse virus (LACV) are orthobunyaviruses that are transmitted by insects and cause meningitis and encephalitis in subsets of individuals in the Americas. Recently, we demonstrated that components of the type I interferon (IFN) induction pathway, particularly the regulatory transcription factors IRF-3 and IRF-7, have key protective roles during OROV infection. However, the lethality in Irf3 −/− Irf7 −/− (DKO) mice infected with OROV was not as rapid or complete as observed in Ifnar −/− mice, indicating that other transcriptional factors associated with an IFN response contribute to antiviral immunity against OROV. Here, we evaluated bunyavirus replication, tissue tropism, and cytokine production in primary cells and mice lacking IRF-5. We demonstrate an important role for IRF-5 in preventing neuroinvasion and the ensuing encephalitis caused by OROV and LACV