Infectious pancreatic necrosis virus (IPNV) recombinant viral protein 1 (VP1) and VP2-Flagellin fusion protein elicit distinct expression profiles of cytokines involved in type 1, type 2, and regulatory T cell response in rainbow trout (Oncorhynchus mykiss).

In this study, we examined the cytokine immune response against two proteins of infectious pancreatic necrosis virus (IPNV) in rainbow trout (Oncorhynchus mykiss), the virion-associated RNA polymerase VP1 and VP2-Flagellin (VP2-Flg) fusion protein. Since VP1 is not a structural protein, we hypothesize it can induce cellular immunity, an essential mechanism of the antiviral response. At the same time, the fusion construction VP2-Flg could be highly immunogenic due to the presence of the flagellin used as an adjuvant. Fish were immunized with the corresponding antigen in Montanide™, and the gene expression of a set of marker genes of Th1, Th2, and the immune regulatory response was quantified in the head kidney of immunized and control fish. Results indicate that VP1 induced upregulation of ifn-γ, il-12p40c, il-4/13a, il-4/13b2, il-10a, and tgf-ß1 in immunized fish. Expression of il-2a did not change in treated fish at the times tested. The antigen-dependent response was analysed by in vitro restimulation of head kidney leukocytes. In this assay, the group of cytokines upregulated after VP1-restimulation was consistent with those upregulated in the head kidney in vivo. Interestingly, VP1 induced il-2a expression after in vitro restimulation. The analysis of sorted lymphocytes showed that the increase of cytokines occurred in CD4-1+ T cells suggesting that Th differentiation happens in response to VP1. This is also consistent with the expression of t-bet and gata3, the master regulators for Th1/Th2 differentiation in the kidneys of immunized animals. A different cytokine expression profile was found after VP2-Flg administration, i.e., upregulation occurs for ifn-γ, il-4/13a, il-10a, and tgf-ß1, while down-regulation was observed in il-4/13b2 and il-2a. The cytokine response was due to flagellin; only the il-2a effect was dependent upon VP2 in the fusion protein. To the best of our knowledge this study reports for the first-time characteristics of the adaptive immune response induced in response to IPNV VP1 and the fusion protein VP2-Flg in fish. VP1 induces cytokines able to trigger the humoral and cell-mediated immune response in rainbow trout. The analysis of the fish response against VP2-Flg revealed the immunogenic properties of Aeromonas salmonicida flagellin, which can be further tested for adjuvanticity. The novel immunogenic effects of VP1 in rainbow trout open new opportunities for further IPNV vaccine development using this viral protein.

Functional Evidence of the Involvement of the Dynein Light Chain DYNLRB2 in Murine Leukemia Virus Infection.

How murine leukemia virus (MLV) travels from the cell membrane to the nucleus and the mechanism for nuclear entry of MLV DNA in dividing cells still remain unclear. It seems likely that the MLV preintegration complex (PIC) interacts with cellular proteins to perform these tasks. We recently published that the microtubule motor cytoplasmic dynein complex and its regulator proteins interact with the MLV PIC at early times of infection, suggesting a functional interaction between the incoming viral particles, the dynein complex, and dynein regulators. To better understand the role of the dynein complex in MLV infection, we performed short hairpin RNA (shRNA) screening of the dynein light chains on MLV infection. We found that silencing of a specific light chain of the cytoplasmic dynein complex, DYNLRB2, reduced the efficiency of infection by MLV reporter viruses without affecting HIV-1 infection. Furthermore, the overexpression of DYNLRB2 increased infection by MLV. We conclude that the DYNLRB2 light chain of the cytoplasmic dynein complex is an important and specific piece of the host machinery needed for MLV infection.IMPORTANCE Retroviruses must reach the chromatin of their host to integrate their viral DNA, but first they must get into the nucleus. The cytoplasm is a crowded environment in which simple diffusion is slow, and thus viruses utilize retrograde transport along the microtubule network, mediated by the dynein complex. Different viruses use different components of this multisubunit complex. We have found that murine leukemia virus (MLV) associates functionally and specifically with the dynein light chain DYNLRB2, which is required for infection. Our study provides more insight into the molecular requirements for retrograde transport of the MLV preintegration complex and demonstrates, for the first time, a role for DYNLRB2 in viral infection.

Electrochemical Deposition of Polypyrrole in the Presence of Silanes as Adhesion Promoters.

Polypyrrole adhesion to indium-tin oxide electrodes was improved by adding pre-hydrolyzed alkoxysilanes to the electrodeposition media. The pyrrole oxidation and film growth rates were studied by potentiostatic polymerization in acidic media. The morphology and thickness of the films were studied by contact profilometry and surface-scanning electron microscopy. The bulk and surface semiquantitative chemical composition was studied by Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy. Finally, the adhesion was studied by scotch-tape adhesion test, where both alkoxysilanes showed a significant improvement in adhesion. We proposed a hypothesis for the improvement in adhesion, that involves the formation of siloxane material as well as in situ surface modification of the transparent metal oxide electrode.

CXCL9-11 chemokines and CXCR3 receptor in teleost fish species.

The coordinated migration of immune cells from lymphoid organs to in or out of the bloodstream, and towards the site of infection or tissue damage is fundamental for an efficient innate and adaptive immune response. Interestingly, an essential part of this movement is mediated by chemoattractant cytokines called chemokines. Although the nature and function of chemokines and their receptors are well documented in mammals, much research is needed to accomplish a similar level of understanding of the role of chemokines in fish immunity. The first chemokine gene identified in teleosts (rainbow trout, Oncorhynchus mykiss) was CK1 in 1998. Since then, the identification of fish chemokine orthologue genes and characterization of their role has been more complex than expected, primarily because of the whole genome duplication processes occurring in fish, and because chemokines evolve faster than other immune genes. Some of the most studied chemokines are CXCL9, CXCL10, CXCL11, and the CXCR3 receptor, all involved in T cell migration and in the induction of the T helper 1 (Th1) immune response. Data from the zebrafish and rainbow trout CXCL9-11/CXCR3 axis suggest that these chemokines and the receptor arose early in evolution and must be present in most teleost fish. However, the pieces of knowledge also indicate that different numbers of gene copies can be present in different species, with distinct regulatory expression mechanisms and probably, also with different roles, as the differential expression in fish tissues suggest. Here, we revised the current knowledge of the CXCL9-11/CXCR3 axis in teleost fishes, identifying the gaps in knowledge, and raising some hypotheses for the role of CXCL9, CXCL10 CXCL11, and CXCR3 receptor axis in fish, which can encourage further studies in the field.

Mordenite-Supported Ag+-Cu2+-Zn2+ Trimetallic System: A Variety of Nanospecies Obtained via Thermal Reduction in Hydrogen Followed by Cooling in an Air or Hydrogen Atmosphere.

Multimetallic systems, instead of monometallic systems, have been used to develop materials with diverse supported species to improve their catalytic, antimicrobial activity, etc., properties. The changes in the types of nanospecies obtained through the thermal reduction of ternary Ag+-Cu2+-Zn2+/mordenite systems in hydrogen, followed by their cooling in an air or hydrogen atmosphere, were studied. Such combinations of trimetallic systems with different metal content, variable ratios (between them), and alternating atmosphere types (during the cooling after reducing the samples in hydrogen at 350 °C) lead to diversity in the obtained copper and silver nanospecies. No reduction of Zn2+ was evidenced. A low silver content leads to the formation of reduced silver clusters, while the formation of nanoparticles of a bigger size takes place in the trimetallic samples with high silver content. The cooling of the reduced trimetallic samples in the air causes the oxidation of the obtained metallic clusters and silver and copper nanoparticles. In the case of copper, such conditions lead to the formation of mainly copper (II) oxide, while the silver nanospecies are converted mainly into clusters and nanoparticles. The zinc cations provoked changes in the mordenite matrix, which was associated with the formation of point oxygen defects in the mordenite structure and the formation of surface zinc oxide sub-nanoparticles in the samples cooled in the air.

Copper-silver bimetallic system on natural clinoptilolite: thermal reduction of Cu2+ and Ag+ exchanged.

Copper-silver bimetallic system supported on natural clinoptilolite from Tasajeras deposit (Cuba) was studied. Bimetallic samples were prepared by simultaneous ion exchange, and reduced in a wide temperature range in a hydrogen flow. The main goal of the work was analysis of the mutual influence of both metals on their reduction process and the properties of the resultant particles. Analysis was done by combined use of XRD and UV-Vis spectroscopy. The reduction of Cu2+ and Ag+ cations shows existence of notable inter-influence between both cations during this process. The Cu2+ reduction is favored by the presence of Ag+, which should be related with the synergetic influence of silver cations and/or clusters formed on the first stages of reduction on Cu(2+)-framework interaction, facilitating the Cu2+ reduction even at low temperature (25 and 50 degrees C). The aggregation of the reduced highly dispersed species both for copper and silver is limited in this bimetallic system. The introduction of Ag+ as the second cation in the copper-exchanged zeolites favors the copper reduction at lower temperatures (25 and 50 degrees C), and appears to be the efficient tool for the control of the size of the resultant reduced nanoparticles (it means their dispersion).

Revisiting the Teleost Thymus: Current Knowledge and Future Perspectives.

The thymus in vertebrates plays a critical role in producing functionally competent T-lymphocytes. Phylogenetically, the thymus emerges early during evolution in jawed cartilaginous fish, and it is usually a bilateral organ placed subcutaneously at the dorsal commissure of the operculum. In this review, we summarize the current understanding of the thymus localization, histology studies, cell composition, and function in teleost fishes. Furthermore, we consider environmental factors that affect thymus development, such as seasonal changes, photoperiod, water temperature fluctuations and hormones. Further analysis of the thymus cell distribution and function will help us understand how key stages for developing functional T cells occur in fish, and how thymus dynamics can be modulated by external factors like photoperiod. Overall, the information presented here helps identify the knowledge gaps and future steps needed for a better understanding of the immunobiology of fish thymus.

General concepts in biostatistics and clinical epidemiology: Random error and systematic error. / Conceptos generales en bioestadística y epidemiología clínica: error aleatorio y error sistemático.

Biomedical research, particularly when it involves human beings, is always subjected to sources of error that must be recognized. Systematic error or bias is associated with problems in the methodological design or during the execu-tion phase of a research project. It affects its validity and is qualitatively ap-praised. On the other hand, random error is related to variations due to chance. It may be quantitatively expressed, but never removed. This review is the first of a methodological series on general concepts in biostatistics and clin-ical epidemiology developed by the Chair of Scientific Research Methodology at the School of Medicine, University of Valparaíso, Chile. In this article, we address the theoretical concepts of error, its evaluation, and control. Finally, we discuss some current controversies in its conceptualization that are relevant to undergraduate and graduate students of health sciences.

La investigación biomédica, particularmente la que involucra a seres humanos, está siempre sometida a fuentes de error que deben ser reconocidas. El error sistemático o sesgo, se asocia con debilidades en el diseño metodológico o de la fase de ejecución del estudio. Éste afecta su validez y se valora cualitativamente. Por su parte, el error aleatorio se relaciona con las variaciones producidas por el azar, el cual puede expresarse cuantitativamente, pero nunca eliminarse. Esta revisión es la primera entrega de una serie metodológica sobre conceptos generales en bioestadística y epidemiología clínica desarrollada por la Cátedra de Metodología de la Investigación Científica de la Universidad de Valparaíso, Chile. En este artículo se abordan los conceptos teóricos asociados al error, su evaluación y control. Finalmente, se discuten algunas controversias actuales en cuanto a su conceptualización, de relevancia para estudiantes de pre y posgrado de ciencias de la salud.

Enhanced photocatalytic degradation of sulfamethoxazole by deposition of Au, Ag and Cu metallic nanoparticles on TiO2.

Mono- (Au, Ag and Cu) and bi-metallic (Au-Ag and Au-Cu) nanoparticles were deposited on TiO2 and tested for the photocatalytic degradation of sulfamethoxazole using either UV-C or simulated sunlight. The optimal loading of metallic nanoparticles was determined as 1.5 wt% for Au and Ag, and 1.0 wt% for Cu. In the case of bi-metallic nanoparticles, only the ratio 1:0.5 wt% for both Au-Ag and Au-Cu was tested. In experiments using UV-C light, the highest degradation performance was found for Ag/TiO2, while bi-metallic nanoparticles supported on TiO2 also showed increased photocatalytic activity compared with unmodified TiO2. In simulated sunlight irradiation tests, Au/TiO2 showed to be the most efficient material. Complete mineralization of sulfamethoxazole was achieved when surface-modified materials were tested in both UV-C and simulated sunlight experiments. Photolysis was efficient to fully degrade sulfamethoxazole, although mineralization was lower than 10% for both luminic sources. The main by-products of sulfamethoxazole were determined in photolysis and photocatalysis tests using UV-C light, and degradation paths were proposed. By-products showed non-toxicity and low antibiotic activity. Reuse of the catalysts upon three reaction cycles did not result in the loss of activity.

Desensibilización y reprocesamiento por movimientos oculares (EMDR) para trastornos ansiosos y depresivos en niños y adolescentes: revisión de la evidencia disponible / Eye movement desensitization and reprocessing (EMDR) for anxiety and depressive disorders in children and adolescents: a review of the available literature

Resumen Introducción: la desensibilización y reprocesamiento por movimientos oculares (EMDR) tiene abundante evidencia de eficacia en desórdenes del espectro traumático. Su eficacia en trastornos ansiosos (TA) y depresivos (TD) en niños, niñas y adolescentes ha sido escasamente estudiada. Método: se realizó una revisión narrativa para describir la evidencia disponible sobre eficacia de EMDR en TA y TD en población infantojuvenil. Se buscaron artículos disponibles en PubMed/Medline, SciELO, PsycINFO y Cochrane Library. Se incluyeron todos los artículos primarios y secundarios que evaluaron el efecto de EMDR en TA y TD en población infantojuvenil. Se revisaron sus referencias como segundo método de inclusión. Resultados: se identificaron nueve estudios (cinco en TA y cuatro en TD); tres fueron observacionales y seis experimentales. Todos tuvieron tamaños muestrales reducidos. En TA, los estudios corroboraron la eficacia de EMDR sobre el temor fóbico en fobia a las arañas, pero no sobre la conducta evitativa, donde sería superior la exposición in vivo. Dos series de casos expusieron la utilidad de EMDR en fagogobia y en TA asociados a epilepsia. En TD, EMDR fue eficaz en la reducción de sintomatología depresiva en el contexto del trastorno depresivo mayor, trastorno de estrés agudo y trastornos conductuales. EMDR fue comparable a terapia cognitivo-conductual. Conclusiones: la evidencia corrobora la eficacia de EMDR en TA y TD en niños, niñas y adolescentes. Sin embargo, es muy escasa y cuenta con limitaciones metodológicas. Es necesario realizar estudios experimentales con protocolos estandarizados y especializados de EMDR para TA y TD en población infantojuvenil.

Introduction: Eye movement desensitization and reprocessing (EMDR) has abundant evidence of efficacy in traumatic spectrum disorders. Its efficacy in anxiety disorders (AD) and depressive disorders (DD) in children and adolescents has been scarcely studied. Methods: We conducted a narrative review to describe the available evidence on the efficacy of EMDR in AD and DD in children and adolescents. We searched for articles available in PubMed/Medline, SciELO, PsycInfo and the Cochrane Library. All primary and secondary studies evaluating the effect of EMDR on AD and DD in children and adolescents were included. Their references were reviewed as a second method of inclusion. Results: nine studies were identified (five in AD and four in DD); three were observational and six experimental. All had small sample sizes. In AD, studies corroborated the efficacy of EMDR on phobic fear in spider phobia, but not on avoidance behavior, where in vivo exposure would be superior. Two case series reported the efficacy of EMDR in choking phobia and AD associated with epilepsy. In DD, EMDR was effective in reducing depressive symptomatology in the context of major depressive disorder, acute stress disorder, and conduct disorders. EMDR was comparable to cognitive behavioral therapy. Conclusions: The evidence corroborates the efficacy of EMDR in AD and DD in children and adolescents. However, it is very scarce and has methodological limitations. It is necessary to carry out experimental studies with standardized and specialized EMDR protocols for AD and DD in the child and adolescent population.

Low Concentration Fe-Doped Alumina Catalysts Using Sol-Gel and Impregnation Methods: The Synthesis, Characterization and Catalytic Performance during the Combustion of Trichloroethylene.

The role of iron in two modes of integration into alumina catalysts was studied at 0.39 wt% Fe and tested in trichloroethylene combustion. One modified alumina was synthesized using the sol-gel method with Fe added in situ during hydrolysis; another modification was performed using calcined alumina, prepared using the sol-gel method and impregnated with Fe. Several characterization techniques were used to study the level of Fe modification in the γ-Al2O3 phase formed and to correlate the catalytic properties during trichloroethylene (TCE) combustion. The introduction of Fe in situ during the sol-gel process influenced the crystallite size, and three iron species were generated, namely, magnetite, maghemite and hematite. The impregnated Fe-alumina formed hematite and maghemite, which were highly dispersed on the γ-Al2O3 surface. The X-ray photoelectron spectra (XPS), FT-IR and Mössbauer spectroscopy analyses revealed how Fe interacted with the γ-Al2O3 lattice in both catalysts. The impregnated Fe-catalyst showed the best catalytic performance compared to the catalyst that was Fe-doped in situ by the sol-gel method; both had better catalytic activity than pure alumina. This difference in activity was correlated with the accessibility of the reactants to the hematite iron species on the surface. The chlorine poisoning for all three catalysts was less than 1.8%.