The choice of alternative 5' splice sites in influenza virus M1 mRNA is regulated by the viral polymerase complex.

The influenza virus M1 mRNA has two alternative 5' splice sites: a distal 5' splice site producing mRNA3 that has the coding potential for 9 amino acids and a proximal 5' splice site producing M2 mRNA encoding the essential M2 ion-channel protein. Only mRNA3 was made in uninfected cells transfected with DNA expressing M1 mRNA. Similarly, using nuclear extracts from uninfected cells, in vitro splicing of M1 mRNA yielded only mRNA3. Only when the mRNA3 5' splice site was inactivated by mutation was M2 mRNA made in uninfected cells and in uninfected cell extracts. In influenza virus-infected cells, M2 mRNA was made, but only after a delay, suggesting that newly synthesized viral gene product(s) were needed to activate the M2 5' splice site. We present strong evidence that these gene products are the complex of the three polymerase proteins, the same complex that functions in the transcription and replication of the viral genome. Gel shift experiments showed that the viral polymerase complex bound to the 5' end of the viral M1 mRNA in a sequence-specific and cap-dependent manner. During in vitro splicing catalyzed by uninfected cell extracts, the binding of the viral polymerase complex blocked the mRNA3 5' splice site, resulting in the switch to the M2 mRNA 5' splice site and the production of M2 mRNA.

Cancer vaccines: the interleukin 2 dosage effect.

Cancer vaccines genetically engineered to produce interleukin 2 have been investigated intensively in a series of animal models and are at the point of entering into clinical trials. In this study we demonstrate a strong correlation between the rate of interleukin 2 production and the protection efficiency of murine S91 melanoma cell (clone M-3) vaccines. Best immunization is achieved with vaccines producing medium interleukin 2 levels of 1000-3000 units per 10(5) cells per day. Reduced interleukin 2 production evokes a corresponding decline in the number of successfully treated animals. Unexpectedly, when interleukin 2 expression is raised to high levels of 5000-7500 units per 10(5) cells per day, protection is completely absent because of impaired generation of tumor-specific cytotoxic T lymphocytes. In comparison, granulocyte-macrophage colony-stimulating factor as immunomodulator induces substantial immunization even at a moderate level of secretion and protects all animals at the maximal obtainable level of secretion. Our findings demonstrate the importance of the interleukin 2 level produced by genetically modified tumor cells and may have substantial impact for the clinical application of cancer vaccines.

Factores asociados a la predisposición a vacunarse contra la gripe pandémica A/H1N1 en población adulta del Departamento de Salud de Elche (España): influencia de las fuentes de información

Fundamento. Evaluar en población general las fuentes de información, actitudes y predisposición hacia la vacunación contra la gripe pandémica A/H1N1 de 2009. Métodos. Estudio descriptivo de carácter transversal realizado entre el 25 de noviembre y 30 de diciembre de 2009 mediante entrevista personal cara a cara a una muestra aleatoria (826) de adultos residentes en el Departamento de Salud de Elche (España). Resultados. Los encuestados manifestaron que la televisión (57%) y el médico de familia (47,9%) eran su fuente principal de información sobre vacunas. El 82,2% tenía una buena opinión sobre las vacunas, un 30,5% percibía la gripe A/H1N1 como más grave que la estacional, siendo esta percepción creciente entre los de mayor edad y con menos estudios. Un 25,4% de encuestados sentía preocupación por padecerla, sobre todo los de menor nivel educativo. Un 42,1% manifiesta su buena predisposición para vacunarse contra la gripe estacional, disminuyendo hasta un 18,4% la intención hacia la gripe A/H1N1. La predisposición hacia la vacunación crece con la edad y en el caso de la gripe A/H1N1 decrece a mayor nivel educativo. El médico de familia es la fuente de información más determinante para inmunizarse frente a gripe estacional (OR 1,43) y gripe A/H1N1 (OR 2,47). Conclusiones. Existe baja aceptabilidad de la vacuna pandémica y baja percepción de gravedad sobre la gripe A/H1N1. La experiencia previa de vacunación ante gripe estacional predispone hacia la inmunización contra gripe A/H1N1. Aunque los medios de comunicación encabezan la fuente de información más usual durante este episodio, la influencia del médico de familia en la decisión de vacunarse resulta significativa.

Acceptability of pandemic A(H1N1) influenza vaccination by Essential Community Workers in 2010 Alicante (Spain), perceived seriousness and sources of information

Objective. Describe acceptability of pandemic A(H1N1) influenza vaccination by Essential Community Workers (ECWs) from Alicante province (Spain) in January 2010. Evaluate the correlation with attitudes, beliefs, professional advice and information broadcasted by media. Method. In this cross-sectional study, face-to-face interviews were conducted with 742 ECWs to assess their attitudes towards vaccination against the pandemic influenza strain. A multivariable regression model was made to adjust the Odds Ratios (ORs). Results. Some ECWs reported having been vaccinated with seasonal vaccine, 21.5% (95%IC 18.6–24.9); only 15.4% (95%IC 12.8–18.4) with the pandemic one. ECWs vaccinated regularly against seasonal flu (OR 5.1; 95%IC 2.9–9.1), those who considered pandemic influenza as a severe or more serious disease than seasonal flu (OR 3.8; 95%IC 2.1–6.7) and those who never had doubts about vaccine safety (OR 3.7; 95%IC2.1–6.7) had a better acceptance of pandemic vaccine. Finally, 78.7% (95%IC 75.1–81.4) had doubts about pandemic vaccine's effectiveness. Conclusion. The vast amount of information provided by the media did not seem to be decisive to prevent doubts or to improve the acceptability of the vaccine in ECWs. Professional advice should be the focus of interest in future influenza vaccination campaigns. These results should be taken into account by health authorities.

Creencias, actitudes e influencia de los medios de comunicación en trabajadores sanitarios españoles durante la gripe pandémica A (H1N1) 2009

Introducción: Considerado como grupo de riesgo específico en la estrategia de inmunización contra la gripe A (H1N1), el colectivo de trabajadores sanitarios ha sido objeto de este estudio desde la perspectiva de sus actitudes y creencias hacia la inmunización, con especial énfasis en la influencia de las fuentes de información para tomar la decisión de vacunarse. Métodos: Estudio observacional de carácter transversal dirigido a trabajadores sanitarios en activo de la provincia de Alicante y realizado mediante cuestionario cara a cara a una muestra aleatoria por cuotas según categoría profesional en trabajadores de hospitales y centros de salud. Resultados: Las fuentes de información difieren entre subgrupos: los médicos utilizaron revistas científicas y/o congresos, las enfermeras la obtuvieron a través de Sanidad y otras enfermeras, el resto de trabajadores optaron por la televisión y/o el médico de familia. De los 3 colectivos estudiados, los médicos son los que menos sensación de gravedad han percibido frente a la gripe A (H1N1) (59,4%), son los que más confían en la vacuna (42,3%), los que más la recomiendan (44,4%), los que mejor han seguido las recomendaciones para evitar el contagio (93%) y los más vacunados (18,3%). El 75,5% de los sanitarios valoró la información recibida como regular, mala o muy mala. La totalidad admitió que se creó alarma social. Discusión: El éxito de futuras campañas de inmunización contra la gripe en personal sanitario podría incrementarse si fueran diseñadas actividades informativas segmentadas y orientadas a cada subgrupo del colectivo, adecuando la estrategia y mejorando la calidad de la información.

Conocimiento y cobertura vacunal autodeclarada en inmigrantes adultos residentes en la provincia de Alicante

Introducción: Existe una estimable probabilidad de cobertura vacunal insuficiente entre la población inmigrante. Los estudios realizados se centran en población infantil. Este trabajo explora la opinión y conocimientos sobre vacunas, así como la cobertura vacunal autodeclarada en población adulta inmigrante en edad laboral. Métodos: Estudio transversal basado en un cuestionario específico dirigido a inmigrantes entre 18-65 años residentes en la provincia de Alicante. Se realizó mediante entrevista personal a una muestra de 692 individuos entre febrero y abril de 2010. Resultados: Del total de encuestados, un 56,6% son mujeres, el 90,8% reside en España desde hace menos de 10 años y un 88,7% dispone de tarjeta sanitaria. Las comunidades rumanas y marroquíes son las que menos confianza muestran hacia las vacunas. Las más conocidas son las del tétanos (65,8%), la gripe (56,8%) y la hepatitis B (56,2%); a su vez, las más administradas son las del tétanos y la hepatitis B (entre los marroquíes), y la antigripal (entre los europeos). El colectivo marroquí es el peor vacunado en su país de origen y el que más vacunas ha recibido en España (1,3 vacunas/persona). Un 46,7% refiere haber sido inmunizado alguna vez en España, aconsejados principalmente en su centro de salud o el lugar de trabajo. Un 13,3% del colectivo rumano y un 4,7% del ecuatoriano declararon haber tenido alguna dificultad para ser vacunados. Conclusiones: Aunque existe una opinión general favorable hacia las vacunas, algunas nacionalidades muestran cierta indiferencia o desapego. El estado vacunal y la predisposición a vacunarse difiere entre nacionalidades. Sería muy conveniente reforzar la tarea realizada por los equipos de atención primaria y aprovechar las ocasiones que representa la visita a un centro sanitario para ampliar la cobertura vacunal del colectivo de inmigrantes adultos.

Effectiveness of influenza vaccination programme in preventing hospital admissions, Valencia, 2014/15 early results

Preliminary results for the 2014/15 season indicate low to null effect of vaccination against influenza A(H3N2)-related disease. As of week 5 2015, there have been 1,136 hospital admissions, 210 were due to influenza and 98% of subtype A strains were H3. Adjusted influenza vaccine effectiveness was 33% (range: 6–53%) overall and 40% (range: 13% to 59%) in those 65 years and older. Vaccination reduced by 44% (28–68%) the probability of admission with influenza.

La «gripe española» según el diario España Médica (1918-1919)

El siglo XX se vio castigado por una pandemia de gripe que azotó el mundo durante 3 oleadas distribuidas entre febrero de 1918 y junio de 1919. La enorme difusión y gravedad de sus casos caracterizaron a la enfermedad, que fue etiquetada con el nombre de «gripe española». El origen de esta denominación está en la ausencia de censura mediática en España, país no contendiente en la Primera Guerra Mundial, lo que propició la libre circulación de noticias sobre la pandemia, que dieron lugar al equívoco. La pandemia puso en evidencia la escasa efectividad de los recursos médicos de la época, pese al apogeo de las nuevas especialidades nacidas en la era bacteriológica. El impacto social y magnitud de la epidemia fueron recogidos, entre otros, por el periódico España Médica. Fundado y dirigido por el pediatra José Ignacio Eleizegui López (1879-1956), el análisis de las noticias de ese periodo aporta una visión sobre las claves del desarrollo de la enfermedad, la gestión administrativa y los recursos terapéuticos y preventivos empleados.

Influenza A virus ribonucleoproteins modulate host recycling by competing with Rab11 effectors

Influenza A virus assembly is an unclear process, whereby individual virion components form an infectious particle. The segmented nature of the influenza A genome imposes a problem to assembly because it requires packaging of eight distinct RNA particles (vRNPs). It also allows genome mixing from distinct parental strains, events associated with influenza pandemic outbreaks. It is important to public health to understand how segmented genomes assemble, a process that is dependent on the transport of components to assembly sites. Previously, it has been shown that vRNPs are carried by recycling endosome vesicles, resulting in a change of Rab11 distribution. Here, we describe that vRNP binding to recycling endosomes impairs recycling endosome function, by competing for Rab11 binding with family-interacting proteins, and that there is a causal relationship between Rab11 ability to recruit family-interacting proteins and Rab11 redistribution. This competition reduces recycling sorting at an unclear step, resulting in clustering of single- and double-membraned vesicles. These morphological changes in Rab11 membranes are indicative of alterations in protein and lipid homeostasis during infection. Vesicular clustering creates hotspots of the vRNPs that need to interact to form an infectious particle.

Hospitalization risk due to respiratory illness associated with genetic variation at IFITM3 in patients with influenza A(H1N1)pdm09 infection: a case-control study

Recent studies suggest an association between the Interferon Inducible Transmembrane 3 (IFITM3) rs12252 variant and the course of influenza infection. However, it is not clear whether the reported association relates to influenza infection severity. The aim of this study was to estimate the hospitalization risk associated with this variant in Influenza Like Illness (ILI) patients during the H1N1 pandemic influenza. A case-control genetic association study was performed, using nasopharyngeal/oropharyngeal swabs collected during the H1N1 pandemic influenza. Laboratory diagnosis of influenza infection was performed by RT-PCR, the IFITM3 rs12252 was genotyped by RFLP and tested for association with hospitalization. Conditional logistic regression was performed to calculate the confounder-adjusted odds ratio of hospitalization associated with IFITM3 rs12252. We selected 312 ILI cases and 624 matched non-hospitalized controls. Within ILI Influenza A(H1N1)pdm09 positive patients, no statistical significant association was found between the variant and the hospitalization risk (Adjusted OR: 0.73 (95%CI: 0.33–1.50)). Regarding ILI Influenza A(H1N1)pdm09 negative patients, CT/CC genotype carriers had a higher risk of being hospitalized than patients with TT genotype (Adjusted OR: 2.54 (95%CI: 1.54–4.19)). The IFITM3 rs12252 variant was associated with respiratory infection hospitalization but not specifically in patients infected with Influenza A(H1N1)pdm09.

Monitoring influenza vaccine effectiveness using the national influenza surveillance system

Background: Flu vaccine composition is reformulated on a yearly basis. As such, the vaccine effectiveness (VE) from previous seasons cannot be considered for subsequent years, and it is necessary to monitor the VE for each season. This study (MonitorEVA- monitoring vaccine effectiveness) intends to evaluate the feasibility of using the national influenza surveillance system (NISS) for monitoring the influenza VE. Material and methods: Data was collected within NISS during 2004 to 2014 seasons. We used a case-control design where laboratory confirmed incident influenza like illness (ILI) patients (cases) were compared to controls (ILI influenza negative). Eligible individuals consisted on all aged individuals that consult a general practitioner or emergency room with ILI symptoms with a swab collected within seven days of symptoms onset. VE was estimated as 1- odds ratio of being vaccinated in cases versus controls adjusted for age and month of onset by logistic regression. Sensitivity analyses were conducted to test possible effect of assumptions on vaccination status, ILI definition and timing of swabs (<3 days after onset). Results: During the 2004-2014 period, a total of 5302 ILI patients were collected but 798 ILI were excluded for not complying with inclusion criteria. After data restriction the sample size in both groups was higher than 148 individuals/ season; minimum sample size needed to detect a VE of at least 50% considering a level of significance of 5% and 80% power. Crude VE point estimates were under 45% in 2004/05, 2005/06, 2011/12 and 2013/14 season; between 50%-70% in 2006/07, 2008/09 and 2010/11 seasons, and above 70% in 2007/08 and 2012/13 season. From season 2006/07 to 2013/14, all crude VE estimates were statistically significant. After adjustment for age group and month of onset, the VE point estimates decreased and only 2008/09, 2012/13 and 2013/14 seasons were significant. Discussion and Conclusions: MonitorEVA was able to provide VE estimates for all seasons, including the pandemic, indicating if the VE was higher than 70% and less than 50%. When comparing with other observational studies, MonitorEVA estimates were comparable but less precise and VE estimates were in accordance with the antigenic match of the circulating virus/ vaccine strains. Given the sensitivity results, we propose a MonitorEVA based on: a) Vaccination status defined independently of number of days between vaccination and symptoms onset; b) use of all ILI data independent of the definition; c) stratification of VE according to time between onset and swab (< 3 and ≥3 days).

Influenza dei fattori di vista nel calcolo delle condizioni di comfort termico in ambienti con impianti radianti

L’obiettivo di questa tesi di Laurea è stato di analizzare gli scambi termici radiativi che si verificano all’interno di una stanza; per eseguire questa analisi si sono calcolati i fattori di vista Fij tra diversi punti (i) definiti tramite una griglia regolare posta all’interno della stanza e le superfici che vi si affacciano (j); tale griglia prevede 25 punti posti a 3 altezze diverse, per un totale di 75 punti. Dalla conoscenza delle temperature superficiali si sono ottenuti i valori di temperatura media radiante nei punti della griglia, e successivamente i valori degli indici di benessere PMV (voto medio previsto) e PPD (percentuale prevista di insoddisfatti), fissando i parametri necessari. Il calcolo dei fattori di vista è stato eseguito tramite il software TRISCO®, dopo averne confrontato i risultati forniti con COMSOL® e dopo aver valutato la correttezza di alcune approssimazioni. Il modello utilizzato nelle simulazioni è costituito da una stanza di 5x5x2,8 m, in cui sono stati fatti variare il numero e la posizione di alcuni elementi caratterizzanti, come finestre e radiatori, oltre a tipologia di elementi scaldanti e posizione e numero di pareti esterne, tramite la variazione delle temperature. In seguito si è analizzata la potenza scambiata per irraggiamento dagli elementi scaldanti, per poter riconoscere gli elementi responsabili dei maggiori contributi; nel caso di pavimento o soffitto radiante si è divisa tale superficie in diverse porzioni per riconoscere le zone della superficie radiante più influenti. I risultati ottenuti mostrano che la presenza di radiatori, finestre o pareti esterne, introducendo elementi di disturbo asimmetrici, forniscono andamenti disuniformi di temperatura e PMV, con l’omogeneità che migliora allontanandosi da tali elementi; impianti a pavimento o soffitto radiante producono risultati decisamente migliori, sia per quanto riguarda i valori assoluti di temperatura e PMV, che nell’omogeneità della relativa distribuzione.

Vaccines against neosporosis: what can we learn from the past studies?

Neospora caninum is an intracellular apicomplexan parasite, which is a leading cause of abortion in cattle; thus neosporosis represents an important veterinary health problem and is of high economic significance. The parasite can infect cattle via trans-placental transmission from an infected cow to its fetus (vertical transmission), or through the oral route via ingestion of food or water contaminated with oocysts that were previously shed with the feces of a canid definitive host (horizontal transmission). Although vaccination was considered a rational strategy to prevent bovine neosporosis, the only commercialized vaccine (Neoguard®) produced ambiguous results with relatively low efficacy, and was recently removed from the market. Therefore, there is a need to develop an efficient vaccine capable of preventing both, the horizontal transmission through infected food or water to a naïve animal as well as the vertical transmission from infected but clinically asymptomatic dams to the fetus. Different vaccine strategies have been investigated, including the use of live attenuated vaccines, killed parasite lysates, total antigens or antigen fractions from killed parasites, and subunit vaccines. The vast majority of experimental studies were performed in mice, and to a certain extent in gerbils, but there is also a large number of investigations that were conducted in cattle and sheep. However, it is difficult to directly compare these studies due to the high variability of the parameters employed. In this review, we will summarize the recent advances made in vaccine development against N. caninum in cattle and in mice and highlight the most important factors, which are likely to influence the degree of protection mediated by vaccination.