The physiologic increase in expression of some type I IFN-inducible genes during pregnancy is not associated with improved disease activity in pregnant patients with rheumatoid arthritis

During pregnancy, most patients with rheumatoid arthritis (RA) experience a spontaneous improvement in their condition. Since type I interferons (IFN) have immunomodulatory properties, we investigated whether type I IFN-inducible genes are upregulated in pregnant patients with RA. Peripheral blood mononuclear cells were evaluated using quantitative real-time polymerase chain reaction for type I IFN-inducible genes (IFI 35, IFI44, IFI44L, IFIT3, OAS1, and Siglec1) in patients with RA and healthy women during and after pregnancy as well as in nonpregnant controls. IFN-alpha and IFN-beta levels in sera of patients and healthy donors were analyzed by enzyme linked immunosorbent assay. It was found that healthy women did not show a change of gene expression levels from the second trimester until postpartum, yet some type I IFN-inducible genes were significantly upregulated in pregnant and postpartum women compared with nonpregnant individuals. In patients with RA, a pronounced upregulation of IFI35 and IFI44 at the second trimester and a peak expression of Siglec1 at the third trimester were observed. Pregnancy levels of IFI35 and IFI44 in patients with RA were higher than those of nonpregnant patients with RA. No significant association of gene expression levels with disease activity was found. In the sera of patients and healthy women, IFN-beta was undetectable and IFN-alpha levels remained stable throughout pregnancy and postpartum. Thus, pregnancy can give rise to an increased expression of type I IFN-inducible genes, reflecting an upregulation of the innate immune system. However, an association of type I IFN-inducible genes with pregnancy induced disease amelioration seems unlikely.

Stimulation-specific contribution of p38 and JNK to IFN-beta gene expression in human macrophages

Induction of interferon-beta (IFN-beta) gene expression is a tightly regulated process, and a plethora of studies identified the signal transduction pathway TANK-binding kinase-1 (TBK-1)/IFN regulatory factor-3 (IRF-3) as essential to the induction of IFN-beta gene expression. Data regarding the role of p38 and JNK are rare, however. We investigated the contribution of these kinases to IFN-beta expression in human macrophages treated with poly(I:C), lipopolysaccharide (LPS), Sendai virus, or vesicular stomatitis virus (VSV). We found that all the stimuli induced IFN-beta mRNA, albeit to a different extent. Whereas LPS and VSV induced the phosphorylation of p38 and JNK, neither poly(I:C) nor Sendai virus led to the detection of phosphospecific signals. When inhibiting p38, a VSV-triggered IFN-beta mRNA response was inhibited, whereas inhibiting JNK suppressed an LPS-triggered response, but only when macrophages were primed with IFN-gamma. Neither poly(I:C)-induced nor Sendai virus-induced IFN-beta mRNA expression was affected when p38 and JNK were inhibited. Collectively, the data show that the contribution of p38 and JNK to the expression of IFN-beta occurs in a stimulation-specific manner in human macrophages.

Dual induction of PKR with E2F-1 and IFN-alpha to enhance gene therapy against hepatocellular carcinoma

Overexpression of the transcription factor E2F-1 induces apoptosis in tumor cells. This apoptotic effect is partly mediated through the induction of the double-stranded RNA-activated protein kinase (PKR). Here, we investigate if agents that upregulate PKR could enhance the apoptotic effect of E2F-1 overexpression in liver tumors. In human hepatocellular carcinoma (HCC) cells (Hep3B, HepG2, Huh7), adenovirus-mediated overexpression of E2F-1 (AdCMV-E2F) transcriptionally increased PKR mRNA. The subsequent increase of total and phosphorylated PKR protein was followed by induction of apoptosis. When AdCMV-E2F was combined with the PKR modifier interferon alpha (IFNalpha), PKR was additionally upregulated and both PKR activation and apoptosis were increased. Subcutaneous xenograft tumors were selectively targeted using an adenoviral vector expressing E2F-1 under the control of the human telomerase reverse transcriptase (hTERT) promoter (AdhTERT-E2F). Weekly systemic administration of AdhTERT-E2F inhibited tumor growth. The tumor suppressive effect of AdhTERT-E2F therapy was further enhanced in combination with IFNalpha.Our results demonstrate that PKR activating agents enhance the anti-tumor effect of E2F-1 overexpression in HCC in-vitro and in-vivo. Hence, modulation of PKR is a potential strategy to increase the efficacy of PKR-dependent anti-tumor therapies.

La estimación de la biodiversidad forestal en el Inventario Forestal Nacional. Aplicación en el IFN-4 en Galicia

Teniendo como punto de partida la Convención de la Diversidad biológica de 1992, desde la segunda mitad del siglo XX las perspectivas en política y gestión medioambiental han dado un giro sin precedentes. Variables intangibles, difíciles de medir, como es la biodiversidad, son cada vez más importantes para la sociedad. En consecuencia, enmarcado en políticas y requerimientos internacionales, desde el Tercer Inventario Forestal Nacional (IFN-3) se está aplicando una nueva metodología en continuo desarrollo que trata de estimar nuevas variables relacionadas con la biodiversidad. El objetivo más importante es poder analizar mediante estos nuevos indicadores la evolución del estado de la biodiversidad forestal de nuestros bosques a lo largo de los sucesivos IFN. Para llevar a cabo este reto, la toma de datos de campo en el inventario se centra principalmente en indicadores relativos a la composición florística y a la estructura de las masas forestales. En este artículo se describe la evolución y el desarrollo de la metodología llevada a cabo hasta el momento para la estimación de la biodiversidad forestal en el IFN. Además, se detallan los resultados derivados de su aplicación en las formaciones forestales más representativas de Galicia. Finalmente, se exponen los nuevos indicadores relacionados con la biodiversidad forestal en los que se está investigando, así como nuevas perspectivas de análisis.

Identificación y caracterización del componente proteico de la especialidad farmacéutica Inmunoferon®

El sistema inmune es el sistema de defensa del organismo involucrado en la protección frente a microorganismos patógenos y neoplasias. Este sistema está formado por una gran variedad de células y moléculas capacitadas para reconocer específicamente estructuras moleculares o antígenos y desarrollar una respuesta inmune que conduce a su eliminación. Sin embargo, en ocasiones esta respuesta puede estar alterada provocando enfermedades derivadas de respuesta insuficiente (inmunodeficiencias, infección, neoplasias), o de respuesta excesiva (alergia, autoinmunidad, rechazo de trasplantes). La estrategia terapéutica utilizada para restaurar el correcto funcionamiento de la respuesta inmune, estimulándola o suprimiéndola, se conoce como inmunomodulación. Para lograr la inmunomodulación se utilizan agentes inmunomoduladores de naturaleza muy variada que incluyen sustancias sintéticas, recombinantes y de origen natural. Dentro de este último grupo cabe destacar a los inmunomoduladores diseñados con el objetivo de estimular mecanismos de inmunidad natural. A este grupo pertenece el fármaco español Inmunoferon®. Se trata de un inmunomodulador oral que ha demostrado capacidad para normalizar la función efectora de las células accesorias y fagocíticas, de las células NK y de los linfocitos T. Simultáneamente, inhibe la producción de TNF-α y modula la producción de otras citoquinas reguladoras (IL-1, IL-2, IL-12, IFN- γ). Se ha empleado en enfermedades diversas, como la hepatitis B crónica, la enfermedad pulmonar obstructiva crónica, la estomatitis aftosa y la inflamación muscular, entre otras. El principio activo de la especialidad Inmunoferon® es una asociación no covalente de polisacárido/proteína absorbida sobre una matriz estabilizante de sulfato y fosfato cálcicos. El polisacárido es un glucomanano de la pared de Candida utilis y el componente proteico procede de semillas no germinadas de ricino (Ricinus communis). Hasta el presente, el desconocimiento total de la naturaleza de este componente proteico ha impedido estudiar y conocer en profundidad la compleja farmacología de este fármaco...

Virus de la hepatitis C: variabiliad en las regiones del CORE y NS5A y su impacto en la infección por el genotipo 1b

La infección por el virus de la hepatitis C (VHC) tiene un gran impacto a nivel mundial, considerándose una de las principales causas de hepatitis crónica, enfermedad hepática terminal y hepatocarcinoma. Hasta la incorporación de los nuevos agentes antivirales, el genotipo 1b era el que tenía tasas más bajas de respuesta viral sostenida (RVS); y a su vez, el de mayor prevalencia en nuestro medio. Lo que ha llevado a la búsqueda de marcadores predictores de respuesta, con el fin de identificar a aquellos pacientes que pudieran beneficiarse del tratamiento. Estudios previos han mostrado que la variabilidad genómica en la región codificante de la proteína C del Core y en la región comprendida entre los aminoácidos 2209 y 2248 (región determinante de la sensibilidad al IFN: ISDR) de la región que codifica la proteína no-­‐estructural 5A (NS5A) están relacionados con la respuesta al tratamiento con biterapia. Es por ello, que el objetivo de nuestro estudio es caracterizar a nivel genómico ambas regiones y analizar su impacto en pacientes con infección crónica por el genotipo 1b. Se analizan, además, otros factores basales implicados en la respuesta: sexo, edad y carga viral basal...

Role of IFN-gamma in immune responses to Candida albicans infections

Candida albicans is the most frequent etiologic agent that causes opportunistic fungal infections called candidiasis, a disease whose systemic manifestation could prove fatal and whose incidence is increasing as a result of an expanding immunocompromised population. Here we review the role of interferon-gamma (IFN-γ) in host protection against invasive candidiasis. This cytokine plays an essential role in both the innate and adaptive arms of the immune response to candidiasis. We focus on recent progress on host-pathogen interactions leading to the production of IFN-γ by host cells. IFN-γ is produced by CD4 Th1, CD8, γδ T, and natural killer (NK) cells, essentially in response to both IL-12 and/or IL-18; more recently, a subset of C. albicans-specific Th17 cells have been described to produce both IL-17 and IFN-γ. IFN-γ plays an important role in the regulation of the immune system as well as in the control of the infectious process, as it is required for optimal activation of phagocytes, collaborates in the generation of protective antibody response, and favors the development of a Th1 protective response.

Diagnóstico de tuberculosis en rumiantes y camélidos: optimización de pruebas de base celular y humoral

La tuberculosis es una zoonosis bacteriana que representa un importante problema sanitario en el hombre y los animales (domésticos y silvestres), siendo en estos Mycobacterium bovis y M. caprae las especies de mayor relevancia. La enfermedad tiene un gran impacto económico ya que produce un descenso de la producción de los animales y restricciones en su movimiento, además de los costes derivados de su erradicación en el ganado bovino. Actualmente, la tuberculosis en otras especies domésticas como la cabra, comienza a tener relevancia en aquellos países con elevado censo de caprino. Por otra parte, los camélidos de Sudamérica (alpacas y llamas principalmente), se emplean cada vez con mayor frecuencia fuera de su hábitat natural como mascotas y para la producción de fibra. Además, son altamente susceptibles a la enfermedad, habiéndose notificado varios brotes de tuberculosis en estos animales en Europa. Los programas de erradicación se basan en una estrategia de “diagnóstico y sacrificio”, de tal modo que los animales positivos a las pruebas diagnósticas se eliminan de la explotación. Las técnicas diagnósticas oficiales en la Unión Europea son la prueba de la intradermotuberculinización (IDTB) y el test de detección de interferón-gamma (IFN-γ), que detectan la respuesta inmune de base celular. Existen, además, pruebas para la detección de anticuerpos, algunas disponibles comercialmente y otras en fase experimental. Con el objetivo de optimizar el diagnóstico de la tuberculosis en rumiantes (ganado bovino y caprino) y en camélidos (llamas y alpacas), se han realizado siete estudios experimentales, distribuidos en cuatro capítulos, diseñados para cumplir los cuatro objetivos de la presente tesis doctoral...

Identificación de subpoblaciones de macrófagos y linfocitos T en pulmones de pacientes fallecidos por influenza A H1N1

La infección por el virus de la influenza es un problema de salud pública por su rápida diseminación y alta morbilidad. Los casos graves de infección por éste virus presentan hipercitocinemia, la cual se ha asociado a una respuesta inmune adquirida desfavorable y un mal pronóstico para el paciente. Se han realizado hasta ahora experimentos en suero de pacientes o en tejido pulmonar en modelos de animales, sin embargo, no se tienen reportes del microambiente en el pulmón de pacientes fallecidos por el virus de la influenza. Objetivo: Determinar las subpoblaciones de macrófagos y linfocitos T y su correlación con el daño tisular en pulmón de pacientes fallecidos por influenza A H1N1 y otras enfermedades respiratorias. Material y Métodos: Se identificó y cuantificó el virus de influenza A H1N1 (pdm)09 e influenza A estacional por qRT-PCR, así como se determinó los niveles de citocinas y marcadores de macrófagos por qRT-PCR (IL-2, IL-4,IL-6, IL- 10, IL-12, IL-17, IL-23, IFN-, TGFβ, TNFα, Arg1, Retnlb e iNOS). Se realizaron tinciones de HyE para la determinación del daño tisular. Por otra parte, se realizaron tinciones de inmunohistoquímica para analizar las poblaciones celulares CD14+, CD206+, CD4+, CD8+, FOXP3+ y citocinas (IL-4, IL-10, IL-17 e IFN-) in situ. Resultados: Se determinó la presencia del virus de la influenza A en 10 muestras, de las cuales 4 fueron H1N1 (pdm)09. Además, se obtuvieron 5 muestras de neumonía por Coccidioides spp., y 6 de neumonías de origen bacteriano. No existe diferencia en el daño causado por el virus de la influenza A H1N1 (pdm)09 e influenza A estacional a nivel histopatológico. Las células CD14+ y CD4+ se encontraron aumentadas para todos los grupos de neumonías sin importar el agente etiológico, excepto CD4 para las neumonías bacterianas. Las células que expresaron Foxp3 solo se encontraron aumentadas en el grupo de coccidioidomicosis y neumonías bacterianas. No se encontró diferencia significativa entre los grupos de estudio para las células CD8+, excepto para las neumonías bacterianas. La expresión génica relativa de IL-6 se encontró aumentada 3000 veces la expresión génica en el grupo de influenza pandémica A H1N1 (pdm)09, mientras en influenza estacional se encontró una disminución de 0.08 veces de su expresión. INOS se encontró con expresión disminuida 0.5 veces para los grupos de influenza pandémica, influenza estacional y coccidioidomicosis. La expresión génica de IL-10 se encontró solamente para el grupo de influenza A H1N1 (pdm)09 (P=0.01). Resistin Like Beta se encontró con expresión disminuida solo para el grupo de influenza A H1N1 (pdm)09. Existe un aumento de células positivas para IL4 en todos los grupos, excepto neumonías bacterianas. No se encontró diferencia significativa de células positivas para IL-10 en los grupos de estudio. Existe un aumento de células positivas para IL-17 en influenza A H1N1p/2009, influenza A y neumonías bacterianas. IFN se encontró aumentado en los grupos de neumonía comparados con el control. Conclusiones: Ambos grupos de neumonías por influenza A se caracterizan por daño tisular y un exacerbado ambiente inflamatorio; solamente CD206 es capaz de diferenciar entre influenza A H1N1(pdm)09 e influenza estacional

The Inadequacy Of The Y-axis Of Growth (sngn) For The Vertical Pattern Assessment In Patients With Sagittal Discrepancies.

The aim of this cephalometric study was to evaluate the influence of the sagittal skeletal pattern on the 'Y-axis of growth' measurement in patients with different malocclusions. Lateral head films from 59 patients (mean age 16y 7m, ranging from 11 to 25 years) were selected after a subjective analysis of 1630 cases. Sample was grouped as follows: Group 1 - class I facial pattern; group 2 - class II facial pattern; and Group 3 - class III facial pattern. Two angular measurements, SNGoGn and SNGn, were taken in order to determine skeletal vertical facial pattern. A logistic regression with errors distributed according to a binomial distribution was used to test the influence of the sagittal relationship (Class I, II, III facial patterns) on vertical diagnostic measurement congruence (SNGoGn and SNGn). RESULTS show that the probability of congruence between the patterns SNGn and SNGoGn was relatively high (70%) for group 1, but for groups II (46%) and III (37%) this congruence was relatively low. The use of SNGn appears to be inappropriate to determine the vertical facial skeletal pattern of patients, due to Gn point shifting throughout sagittal discrepancies. Clinical Significance: Facial pattern determined by SNGn must be considered carefully, especially when severe sagittal discrepancies are present.

Effect Of Roux-en-y Gastric Bypass On Nonalcoholic Fatty Liver Disease Evaluated Through Nafld Fibrosis Score: A Prospective Study.

Nonalcoholic fatty liver disease (NAFLD) is common among subjects who undergo bariatric surgery and its postsurgical improvement has been reported. This study aimed to determine the evolution of liver disease evaluated through NAFLD fibrosis score 12 months after surgery. It is a prospective cohort study which evaluated patients immediately before and 12 months following Roux-en-Y gastric bypass (RYGB). Mean score decreased from 1.142 to 0.066; surgery led to a resolution rate of advanced fibrosis of 55 %. Resolution was statistically associated with female gender, percentage of excess weight loss, postsurgical body mass index, postsurgical platelet count, and diabetes resolution. As previously reported by studies in which postsurgical biopsies were performed, RYGB leads to a great resolution rate of liver fibrosis. Since postsurgical biopsy is not widely available and has a significant risk, calculation of NAFLD fibrosis score is a simple tool to evaluate this evolution through a noninvasive approach.

Predictors For Weight Loss Failure Following Roux-en-y Gastric Bypass.

Weight loss failure is a widely recognized occurrence following Roux-en-Y gastric bypass. This study aims to identify predictors associated with weight loss failure. It is a retrospective cohort which enrolled 187 subjects who underwent RYGB. Comparisons were made between patients' features at baseline and 24 months after surgery. A weight loss failure rate of 11.2% was found. Advanced age and diabetes were statistically associated with failure. The results found were close to previous reports. As weight loss failure represents an important concern, there is the possibility to perform revisional surgeries, which may emphasize the restrictive or malabsorptive characteristics of RYGB, leading to varied results. It is reinforced that weight loss cannot be used as the unique outcome to evaluate the success of surgery.

Oropouche Virus Infection And Pathogenesis Is Restricted By Mavs, Irf-3, Irf-7, And Type I Ifn Signaling Pathways In Non-myeloid Cells.

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), IFN-β, 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. In mice, deletion of IFNAR, MAVS, or IRF-3 and IRF-7 resulted in uncontrolled OROV replication, hypercytokinemia, extensive liver damage, and death whereas wild-type (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 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 virus infection and tissue injury, and suggest that IFN signaling in non-myeloid cells contributes to the host defense against orthobunyaviruses. Oropouche virus (OROV) is an emerging arthropod-transmitted orthobunyavirus that causes episodic outbreaks of a debilitating febrile illness in humans in countries of South and Central America. The continued expansion of the range and number of its arthropod vectors increases the likelihood that OROV will spread into new regions. At present, the pathogenesis of OROV in humans or other vertebrate animals remains poorly understood. To define cellular mechanisms of control of OROV infection, we performed infection studies in a series of primary cells and mice that were deficient in key innate immune genes involved in pathogen recognition and control. Our results establish that a MAVS-dependent type I IFN signaling pathway has a dominant role in restricting OROV infection and pathogenesis in vivo.

Resonancias discursivas y cortesía en prácticas de lecto-escritura

In this paper we present a study of reading comprehension based on a contrastive argumentative-discursive approach. We examine the relationship between linguistic materiality and discursive processes, observing the connection between reading in a foreign language, writing production and textual memories in the mother tongue. In addition to an interest in practical language teaching and learning processes (in this case of Spanish and Portuguese), we investigate the question of politeness and the theoretical relationship between subjectivity, language, and textuality. The latter, being understood as the result of discourse regularities, is unique for each and every production, yet is also conditioned by plural discursive memories resulting from contradictory social relationships in a specific historical context (Foucault, 1986; Pêcheux, 1990). In the experiment presented here, we follow some of the procedures of the methodology applied in the European Galatea Project developed for the study of reading strategies in the inter-comprehension between Romance languages (Dabène, 1996). We use the procedure of simulation and the subjective projection of participants as well as the notion of discursive resonance in the analysis. The results, having to do with directness and indirectness in speech and the question of politeness in two typologically close languages, lead to the conclusion that the concept of politeness goes beyond a pragmatic strategy used to avoid conflicts to be approached as a marker of cultural identity constitution. The relevance of discursive awareness and its theoretical and practical consequences are then emphasized.

The recurrence of leprosy reactional episodes could be associated with oral chronic infections and expression of serum IL-1, TNF-α, IL-6, IFN-γ and IL-10

The aim of this study was to determine whether the presence of leprosy reactional episodes could be associated with chronic oral infection. Thirty-eight leprosy patients were selected and divided into 2 groups: group I - 19 leprosy patients with oral infections, and group II - 19 leprosy patients without oral infections. Ten patients without leprosy, but presenting oral infections, were assigned to the control group. Leprosy patients were classified according to Ridley and Jopling classification and reactional episodes of the erythema nodosum type or reversal reaction were identified by clinical and histopathological features associated with serum IL-1, TNF-α, IL-6, IFN-γ and IL-10 levels. These analyses were performed immediately before and 7 days after the oral infection elimination. Patients from group I presenting oral infections reported clinical improvement of the symptoms of reactional episodes after dental treatment. Serum IL-1, TNF-α, IL-6, IFN-γ and IL-10 levels did not differ significantly before and after dental treatment as determined by the Wilcoxon test (p>0.05). Comparison of the 2 groups showed statistically significant differences in IL-1 and IL-6 at baseline and in IL-1, IL-6 and IL-10 on the occasion of both collections 7 days after therapy. Serum IL-6 and IL-10 levels in group I differed significantly at baseline compared to control (Mann-Whitney test; p<0.05). These results suggest that oral infection could be involved as a maintenance factor in the pathogenesis of leprosy reactional episodes.