Infection of human amniotic and endothelial cells by Japanese encephalitis virus: Increased expression of HLA-F

Productive infection of human amniotic and endothelial cell lines with Japanese encephalitis virus (JEV) was established leading to the induction of NF kappa B and HLA-F, a non-classical MHC molecule. Induction of the HLA-F gene and protein in JEV-infected cells was shown to be NF kappa B dependent since it was blocked by inhibitors of NF kappa B activation. ShRNA targeting lentivirus-mediated stable knockdown of the p65 subunit of NF kappa B inhibited JEV-mediated induction of HLA-F both in the amniotic cell line, AV-3 as well as the human brain microendothelial cell line, HBMEC. The induction of HLA-F by treatment of AV-3 with TNF-alpha was also inhibited by ShRNA mediated knockdown of NF kappa B. TNF-alpha treatment of HEK293T cells that were transfected with reporter plasmids under the control of HLA-F enhancer A elements resulted in significant transactivation of the luciferase reporter gene. NF kappa B-mediated induction of HLA-F following JEV infection and TNF-alpha exposure is being suggested for the first time. (C) 2014 Elsevier Inc. All rights reserved.

Long-Term Sustained Release of Salicylic Acid from Cross-Linked Biodegradable Polyester Induces a Reduced Foreign Body Response in Mice

There has been a continuous surge toward developing new biopolymers that exhibit better in vivo biocompatibility properties in terms of demonstrating a reduced foreign body response (FBR). One approach to mitigate the undesired FBR is to develop an implant capable of releasing anti-inflammatory molecules in a sustained manner over a long time period. Implants causing inflammation are also more susceptible to infection. In this article, the in vivo biocompatibility of a novel, biodegradable salicylic acid releasing polyester (SAP) has been investigated by subcutaneous implantation in a mouse model. The tissue response to SAP was compared with that of a widely used biodegradable polymer, poly(lactic acid-co-glycolic acid) (PLGA), as a control over three time points: 2, 4, and 16 weeks postimplantation. A long-term in vitro study illustrates a continuous, linear (zero order) release of salicylic acid with a cumulative mass percent release rate of 7.34 x 10(-4) h(-1) over similar to 1.5-17 months. On the basis of physicochemical analysis, surface erosion for SAP and bulk erosion for PLGA have been confirmed as their dominant degradation modes in vivo. On the basis of the histomorphometrical analysis of inflammatory cell densities and collagen distribution as well as quantification of proinflammatory cytokine levels (TNF-alpha and IL-1 beta), a reduced foreign body response toward SAP with respect to that generated by PLGA has been unambiguously established. The favorable in vivo tissue response to SAP, as manifest from the uniform and well-vascularized encapsulation around the implant, is consistent with the decrease in inflammatory cell density and increase in angiogenesis with time. The above observations, together with the demonstration of long-term and sustained release of salicylic acid, establish the potential use of SAP for applications in improved matrices for tissue engineering and chronic wound healing.

Absence of systemic toxicity in mouse model towards BaTiO3 nanoparticulate based eluate treatment

One of the existing issues in implant failure of orthopedic biomaterials is the toxicity induced by the fine particles released during long term use in vivo, leading to acute inflammatory response. In developing a new class of piezobiocomposite to mimic the integrated electrical and mechanical properties of bone, bone-mimicking physical properties as well as in vitro cytocompatibility properties have been achieved with spark plasma sintered hydroxyapatite (HA)-barium titanate (BaTiO3) composites. However, the presence of BaTiO3 remains a concern towards the potential toxicity effect. To address this issue, present work reports the first result to conclusively confirm the non-toxic effect of HA-BaTiO3 piezobiocomposite nanoparticulates, in vivo. Twenty BALB/c mice were intraarticularly injected at their right knee joints with different concentrations of HA-BaTiO3 composite of up to 25 mg/ml. The histopathological examination confirmed the absence of any trace of injected particles or any sign of inflammatory reaction in the vital organs, such as heart, spleen, kidney and liver at 7 days post-exposure period. Rather, the injected nanoparticulates were found to be agglomerated in the vicinity of the knee joint, surrounded by macrophages. Importantly, the absence of any systemic toxicity response in any of the vital organs in the treated mouse model, other than a mild local response at the site of delivery, was recorded. The serum biochemical analyses using proinflammatory cytokines (TNF-alpha and IL-1 beta) also complimented to the non-immunogenic response to injected particulates. Altogether, the absence of any inflammatory/ adverse reaction will open up myriad of opportunities for BaTiO3 based piezoelectric implantable devices in biomedical applications.

Sonic hedgehog-Dependent Induction of MicroRNA 31 and MicroRNA 150 Regulates Mycobacterium bovis BCG-Driven Toll-Like Receptor 2 Signaling

Hedgehog (HH) signaling is a significant regulator of cell fate decisions during embryogenesis, development, and perpetuation of various disease conditions. Testing whether pathogen-specific HH signaling promotes unique innate recognition of intracellular bacteria, we demonstrate that among diverse Gram-positive or Gram-negative microbes, Mycobacterium bovis BCG, a vaccine strain, elicits a robust activation of Sonic HH (SHH) signaling in macrophages. Interestingly, sustained tumor necrosis factor alpha (TNF-alpha) secretion by macrophages was essential for robust SHH activation, as TNF-alpha(-/-) macrophages exhibited compromised ability to activate SHH signaling. Neutralization of TNF-alpha or blockade of TNF-alpha receptor signaling significantly reduced the infection-induced SHH signaling activation both in vitro and in vivo. Intriguingly, activated SHH signaling downregulated M. bovis BCG-mediated Toll-like receptor 2 (TLR2) signaling events to regulate a battery of genes associated with divergent functions of M1/M2 macrophages. Genome-wide expression profiling as well as conventional gain-of-function or loss-of-function analysis showed that SHH signaling-responsive microRNA 31 (miR-31) and miR-150 target MyD88, an adaptor protein of TLR2 signaling, thus leading to suppression of TLR2 responses. SHH signaling signatures could be detected in vivo in tuberculosis patients and M. bovis BCG-challenged mice. Collectively, these investigations identify SHH signaling to be what we believe is one of the significant regulators of host-pathogen interactions.

Ac2PIM-responsive miR-150 and miR-143 Target Receptor-interacting Protein Kinase 2 and Transforming Growth Factor Beta-activated Kinase 1 to Suppress NOD2-induced Immunomodulators

Specific and coordinated regulation of innate immune receptor-driven signaling networks often determines the net outcome of the immune responses. Here, we investigated the cross-regulation of toll-like receptor (TLR)2 and nucleotide-binding oligomerization domain (NOD)2 pathways mediated by Ac2PIM, a tetra-acylated form of mycobacterial cell wall component and muramyl dipeptide (MDP), a peptidoglycan derivative respectively. While Ac2PIM treatment of macrophages compromised their ability to induce NOD2-dependent immunomodulators like cyclooxygenase (COX)-2, suppressor of cytokine signaling (SOCS)-3, and matrix metalloproteinase (MMP)-9, no change in the NOD2-responsive NO, TNF-alpha, VEGF-A, and IL-12 levels was observed. Further, genome-wide microRNA expression profiling identified Ac2PIM-responsive miR-150 and miR-143 to target NOD2 signaling adaptors, RIP2 and TAK1, respectively. Interestingly, Ac2PIM was found to activate the SRC-FAK-PYK2-CREB cascade via TLR2 to recruit CBP/P300 at the promoters of miR-150 and miR-143 and epigenetically induce their expression. Loss-of-function studies utilizing specific miRNA inhibitors establish that Ac2PIM, via the miRNAs, abrogate NOD2-induced PI3K-PKC delta-MAPK pathway to suppress beta-catenin-mediated expression of COX-2, SOCS-3, and MMP-9. Our investigation has thus underscored the negative regulatory role of Ac2PIM-TLR2 signaling on NOD2 pathway which could broaden our understanding on vaccine potential or adjuvant utilities of Ac2PIM and/or MDP.

Neonatal exposure to 17 beta-estradiol down-regulates the expression of synaptogenesis related genes in selected brain regions of adult female rats

Aims: Administration of estradiol or compounds with estrogenic activity to newborn female rats results in irreversible masculinization as well as defeminization in the brain and the animals exhibit altered reproductive behavior as adults. The cellular and molecular mechanism involved in inducing the irreversible changes is largely unknown. In the present study, we have monitored the changes in the expression of selected synaptogenesis related genes in the sexually dimorphic brain regions such as POA, hypothalamus and pituitary following 17 beta-estradiol administration to neonatal female rats. Main methods: Female Wistar rats which were administered 17 beta-estradiol on day 2 and 3 after birth were sacrificed 120 days later and the expression levels of genes implicated in synaptogenesis were monitored by semi-quantitative reverse transcription PCR. Since estradiol induced up-regulation of COX-2 in POA is a marker for estradiol induced masculinization as well as defeminization, in the present study only animals in which the increase in expression of COX-2 gene was observed in POA were included in the study. Key findings: Down-regulation of genes such as NMDA-2B, NETRIN-1, BDNF, MT-5 MMP and TNF-alpha was observed in the pre-optic area of neonatally E2 treated female rat brain but not in hypothalamus and pituitary compared to the vehicle- treated controls as assessed by RT-PCR and Western blot analysis. Significance: Our results suggest a possibility that down-regulation of genes associated with synaptogenesis in POA, may be resulting in disruption of the cyclical regulation of hormone secretion by pituitary the consequence of which could be infertility and altered reproductive behavior. (C) 2015 Elsevier Inc. All rights reserved.

Molecular profiling of sepsis in mice using Fourier Transform Infrared Microspectroscopy

Sepsis is a life threatening condition resulting from a high burden of infection. It is a major health care problem and associated with inflammation, organ dysfunction and significant mortality. However, proper understanding and delineating the changes that occur during this complex condition remains a challenge. A comparative study involving intra-peritoneal injection of BALB/c mice with Salmonella Typhimurium (infection), lipopolysaccharide (endotoxic shock) or thioglycollate (sterile peritonitis) was performed. The changes in organs and sera were profiled using immunological assays and Fourier Transform Infrared (FTIR) micro-spectroscopy. There is a rapid rise in inflammatory cytokines accompanied with lowering of temperature, respiratory rate and glucose amounts in mice injected with S. Typhimurium or lipopolysaccharide. FTIR identifies distinct changes in liver and sera: decrease in glycogen and protein/lipid ratio and increase in DNA and cholesteryl esters. These changes were distinct from the pattern observed in mice treated with thioglycollate and the differences in the data obtained between the three models are discussed. The combination of FTIR spectroscopy and other biomarkers will be valuable in monitoring molecular changes during sepsis. GRAPHICS] Intra-peritoneal infection with high dose of Salmonella Typhimurium leads to rapid increase in inflammatory cytokines, e.g. Tnf alpha (A). FTIR analysis of liver (B) and sera (C) identifies several metabolic changes: glycogen, protein/lipid, cholesteryl esters and DNA.

The Secreted Protein Rv1860 of Mycobacterium tuberculosis Stimulates Human Polyfunctional CD8(+) T Cells

We previously reported that Rv1860 protein from Mycobacterium tuberculosis stimulated CD4(+) and CD8(+) T cells secreting gamma interferon (IFN-gamma) in healthy purified protein derivative (PPD)-positive individuals and protected guinea pigs immunized with a DNA vaccine and a recombinant poxvirus expressing Rv1860 from a challenge with virulent M. tuberculosis. We now show Rv1860-specific polyfunctional T (PFT) cell responses in the blood of healthy latently M. tuberculosis-infected individuals dominated by CD8(+) T cells, using a panel of 32 overlapping peptides spanning the length of Rv1860. Multiple subsets of CD8(+) PFT cells were significantly more numerous in healthy latently infected volunteers (HV) than in tuberculosis (TB) patients (PAT). The responses of peripheral blood mononuclear cells (PBMC) from PAT to the peptides of Rv1860 were dominated by tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) secretions, the former coming predominantly from non-T cell sources. Notably, the pattern of the T cell response to Rv1860 was distinctly different from those of the widely studied M. tuberculosis antigens ESAT-6, CFP-10, Ag85A, and Ag85B, which elicited CD4(+) T cell-dominated responses as previously reported in other cohorts. We further identified a peptide spanning amino acids 21 to 39 of the Rv1860 protein with the potential to distinguish latent TB infection from disease due to its ability to stimulate differential cytokine signatures in HV and PAT. We suggest that a TB vaccine carrying these and other CD8(+) T-cell-stimulating antigens has the potential to prevent progression of latent M. tuberculosis infection to TB disease.

Inclusión de desperdicios de cocina y galletas en la alimentación de cerdos de engorde

El presente estudio se realizó en la Granja Porcina de la Universidad Nacional Agraria (UNA), ubicada en el kilómetro 13, Carretera Norte, desvió a Sabana Grande, Managua, Nicaragua La realización del ensayó duró 12 semanas comprendidas entre los meses de septiembre-y diciembre de 1998 El objetivo del mismo fue evaluar el comportamiento productivo de cerdos en crecimiento, desarrollo y engorde al utilizar diferentes raciones: concentrado (T1) desperdicios de cocina (T2) y desperdicios de cocina más desperdicios de galleta (T3) Se utilizaron 24 cerdos- híbridos formándose grupos de 8 cerdos por tratamiento, (4 hembras y 4 machos) Los cerdos se pesaron al inicio y luego cada 28 días hasta el final del experimentó, Con las ganancias de peso y conversión alimenticia de los diferentes lotes de cerdos, se efectuó el análisis estadístico correspondiente al diseño experimental completamente al azar (DCA). Los cerdos que presentaron una ganancia de peso más alta, fueron los que se alimentaron con la ración formulada con fuente comercial (tratamiento l) dando una ganancia de peso promedio de 0.693 kg; Los cerdos alimentados con l00% de desperdicios de cocina alcanzaron una ganancia de peso promedio de 0.540 kg, mientras que los del tratamiento 3 alimentados con 50.% con desperdicios de cocina más 50% de desperdicios de galleta alcanzaron una ganancia media de peso promedio de 0,629 kg. El análisis de la ganancia media diaria mostró que hubo diferencia significativa con un alfa de 0.05% entre el tratamiento l y tratamiento 2 y entre los tratamientos 2 y 3, no mostrando diferencia entre los tratamientos 1 y 3, obteniendo el tratamiento 1 la rmejor ganancia media diaria. La conversión de los alimentados bajo el tratamiento de concentrado comercial, fue la más eficiente con un valor de 3.804 Los cerdos bajo el tratamiento 2 presentaron una conversión de 9.691 y el lote de cerdos en el tratamiento 3 presentó una conversión de 7.417. El análisis estadístico mostró que las diferencias en conversión alimenticia son altamente significativas entre el tratamiento l y tratamiento 2 y el tratamiento 1 y el tratamiento 3 y no significativas entre el tratamiento 2 y 3, obteniendo mejor resultado el tratamiento I (concentrado) a un alfa de 0.05%. En la estimación de los costos de los alimentos, el tratamiento 2 produjo los mayores beneficios económicos; debido al bajo costo del alimento con un valor de C$ 9, 856.

Comportamiento productivo de cerdos alimentados con desperdicios de cocina y residuos de galleta en la dieta, bajo distintos porcentajes de inclusión

El presente estudio se realizó en la granja porcina de la Universidad Nacional Agraria (U.N.A), ubicada en el Km. 13 a los 86°. 09' 36" longitud oeste y los 12° 08'15" latitud norte de la comunidad de Sabana Grande, Municipio de Managua con un elevación de 56 m sobre el nivel del mar (INETER, 1987). La fase de campo fue realizada de Julio a Octubre de 1999, con el objetivo de evaluar el comportamiento productivo de cerdos en crecimiento, desarrollo y engorde alimentados con tres raciones diferentes: (T1) Desperdicio de Galleta 100%, (T2) Desperdicio de Galleta 75% + Desperdicio de cocina 25%, (T3) Desperdicio de cocina 100%. Se utilizaron 18 cerdos comerciales de ambos sexos, con peso promedio de inicio de 54.86 kg, que fueron distribuidos en tres tratamientos con seis repeticiones con los datos levantados de pesaje de los diferentes tratamientos, se efectúo el análisis estadístico correspondiente al diseño experimental completamente al Azar (DCA) y su correspondiente análisis para las variables en estudio, G.M D. y C.A. Los cerdos que presentaron una ganancia de peso más alta, fueron los que se alimentaron con la ración l00 % de desperdicio de cocina teniendo una ganancia media diaria de 0.758 kg. los cerdos alimentados 75 % DDG, + 25 % DDC, alcanzaron una ganancia de peso promedio de 0.578 kg. mientras que el tratamiento 100% DDG alcanzo una ganancia media de peso promedio de 0.17 kg. El análisis de la ganancia media diaria se demostró con una alfa de 5 %. El análisis estadístico mostró que las diferencias en conversión alimenticia fueron significativas entre tratamientos con un Alfa al 5 %. Al comparar las utilidades generadas entre el T t y T2, estos resultaron financieramente aceptables, de forma similar se comportaron el T1 y T3., no obstante, desde el punto de vista nutricional, estos no cumplieron los requerimientos de la categoría, por lo que el T2, se presentó finalmente como el más aceptable nutricionalmente.

A Salmonella Typhimurium-Typhi genomic chimera: a model to study Vi polysaccharide capsule function in vivo.

The Vi capsular polysaccharide is a virulence-associated factor expressed by Salmonella enterica serotype Typhi but absent from virtually all other Salmonella serotypes. In order to study this determinant in vivo, we characterised a Vi-positive S. Typhimurium (C5.507 Vi(+)), harbouring the Salmonella pathogenicity island (SPI)-7, which encodes the Vi locus. S. Typhimurium C5.507 Vi(+) colonised and persisted in mice at similar levels compared to the parent strain, S. Typhimurium C5. However, the innate immune response to infection with C5.507 Vi(+) and SGB1, an isogenic derivative not expressing Vi, differed markedly. Infection with C5.507 Vi(+) resulted in a significant reduction in cellular trafficking of innate immune cells, including PMN and NK cells, compared to SGB1 Vi(-) infected animals. C5.507 Vi(+) infection stimulated reduced numbers of TNF-α, MIP-2 and perforin producing cells compared to SGB1 Vi(-). The modulating effect associated with Vi was not observed in MyD88(-/-) and was reduced in TLR4(-/-) mice. The presence of the Vi capsule also correlated with induction of the anti-inflammatory cytokine IL-10 in vivo, a factor that impacted on chemotaxis and the activation of immune cells in vitro.

Modelización de una prueba de afecto hacia la matemática con la teoría de respuesta al ítem

Resumen: Se aplicó el Modelo de Crédito Parcial (MCP) de la Teoría de Respuesta al Ítem (TRI) al análisis de ítems de una escala que mide Afecto hacia la Matemática. Esta variable describe el interés de los estudiantes de Psicología por involucrarse en actividades vinculadas a la matemática y los sentimientos asociados al uso de sus conceptos. La prueba consta de 8 ítems con formato de respuesta Likert de 6 opciones. Participaron 1875 estudiantes de Psicología de la Universidad de Buenos Aires (Argentina) de los cuales un 82% fueron mujeres. El análisis de la consistencia interna brindó un índice altamente satisfactorio (Alfa = .91). Se verificó la condición de unidimensionalidad requerida por el modelo mediante un análisis factorial exploratorio. Todos los análisis basados sobre la TRI se realizaron con el programa Winsteps. La estimación de los parámetros del modelo se efectuó por Máxima Verosimilitud Conjunta. El ajuste del MCP fue satisfactorio para todos los ítems. La Función de Información del Test fue elevada en un rango amplio de niveles del rasgo latente. Un ítem presentó una inversión en dos parámetros de umbral. Como consecuencia, 1 de las 6 categorías del ítem no fue máximamente probable en ningún intervalo de la escala del rasgo latente. Se analizan las implicancias de este hallazgo en la evaluación de la calidad psicométrica del ítem. Los resultados de este estudio permitieron profundizar el análisis del constructo y aportaron evidencias de validez basadas en las estructura interna de la escala

Influencia del tamaño-forma de la parcela experimental y el número de repeticiones sobre la precisión de los datos experimentales, en el cultivo del maíz (Zea mays L.)

Con el objetivo de estudiar la influencia del tamaño y forma de la parcela experimental y el número de repeticiones sobre la precisión de los datos experimentales en el cultivo del maíz (zea mays L) se estableció un Ensayo de Uniformidad en la finca “El plantel “ con la variedad NB-6. El tamaño de la U.B. Los datos del ensayo de uniformidad se analizaron basados en la Ley de varianza de Smith. Siguiendo el procedimiento de Koch y Rignev (1951), para determinar las varianzas correspondientes: El Método de Hatheway (1961), se utilizó para determinar la relación objeto de estudio. Se determinó que: En suelos de heterogeneidad media (b=0.55). Asumiendo un alfa de 5% se pueden emplear combinaciones de tamaños de parcelas con número de repeticiones de: 27.98m2; 8.04m2, 3.87m2 y 2.31m2, con 2, 4, 6 y 8 repeticiones respectivamente: la relación tamaño-forma de la parcela experimental sobre la precisión de los datos obtenidos, no ejerce una influencia relevante o considerable sobre la precisión alcanzada.

Influencia del tamaño-forma de la parcela experimental y el número de repeticiones sobre la precisión de los datos experimentales, en el cultivo de la soya (Glycine max L.)

Con el objetivo de estudiar la influencia del tamaño y forma de la parcela experimental y el número de repeticiones sobre la precisión de los datos experimentales en el cultivo de la soya (Glycine max l) se estableció un Ensayo de Uniformidad en el Centro Experimental del Algodón (C.E.A.) con la variedad Cristalina: El tamaño de la U. B fue de 1.00m2, teniéndose un total de 576 U. B. Los datos del ensayo de uniformidad se analizaron basados en la Ley de varianza de Smith, siguiendo el procedimiento de Koch y Rigney (1951), para determinar las varianza correspondiente; El método de Hatheway y Willams (1958), se utilizo para determinar el coeficiente de heterogeneidad del suelo y el método Harheway (1961), se utilizó para determinar la relación de objeto de estudio. Se determinó que en suelos de heterogeneidad media (b00.41), asumiendo un alfa de 5%, P= 0.8 Gle= 15 y un grado de precisión del 25% se puede emplear tamaños de parcelas comprendidas del 25% se pueden emplear tamaños de parcelas corres prendidas en el rango de 80m2 con 4 repeticiones y se requiere parcelas menores de 10m2 con 6 y 8 repeticiones: También quedó establecido que la relación tamaño- forma de la parcela experimental no ejerce influencia relevante en la precisión de los datos obtenidos.

Role of Monoubiquitylation on the Control of IκBα Degradation and NF-κB Activity

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