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ObjectivesTo analyze the stability of red blood cells, platelets, and reticulocytes of the research parameters, in combination with the respective conventional parameters, for each analyte; and to quantify the morphological changes in these analytes, to propose a correction factor for each.MethodsEthylenediaminetetraacetic acid (EDTA) blood specimens from patients were reanalyzed in 2-hour intervals and then, the mean percentage (X¯t%) changes were calculated. To evaluate the stability of the analyzed material, we used different criteria according to within-run and between-batch analytical variation, as well as intraindividual biological variation. Next, the mean deviation percentage of the parameters that undergo time-dependent significant changes was calculated, to obtain a correction factor.ResultsSeveral conventional and research parameters showed significant alterations in the stability at an early time after arrival at the laboratory.ConclusionCell variations over time can be quantified and corrected by applying a multiplying factor to the signal obtained in the analyzer.

Not available.

Changes have been made to the AnaConDa device (Sedana Medical, Stockholm, Sweden), decreasing its size to reduce dead space and carbon dioxide (CO2) retention. However, this also involves a decrease in the surface area of the activated carbon filter. The CO2 elimination and sevoflurane (SEV) reflection of the old device (ACD-100) were thus compared with the new version (ACD-50) in patients sedated after coronary artery bypass graft surgery. After ERC approval and written informed consent, 23 patients were sedated with SEV, using first the ACD-100 and then the ACD-50 for 60 min each. With each device, patients were ventilated with tidal volumes (TV) of 5 ml/kg of ideal body weight for the first 30 min, and with 7 ml/kg for the next 30 min. Ventilation parameters, arterial blood gases, Bispectral-Index™ (BIS, Aspect Medical Systems Inc., Newton, MA, USA), SEV concentrations exhaled by the patient (SEV-exhaled) and from the expiratory hose (SEV-lost) were recorded every 30 min. A SEV reflection index was calculated: SRI [%] = 100 × (1 − (SEV-lost/SEV-exhaled)). Data were compared using ANOVA with repeated measurements and Student’s T-tests for pairs. Respiratory rates, tidal and minute volumes were not significantly different between the two devices. End tidal and arterial CO2 partial pressures were significantly higher with the ACD-100 as compared with the ACD-50. SEV infusion rate remained constant. SEV reflection was higher (SRI: ACD-100 vs. ACD-50, TV 5 ml/kg: 95.29 ± 6.45 vs. 85.54 ± 11.15, p = 0.001; 7 ml/kg: 93.42 ± 6.55 vs. 88.77 ± 12.26, p = 0.003). BIS was significantly lower when using the higher TV (60.91 ± 9.99 vs. 66.57 ± 8.22, p = 0.012), although this difference was not clinically relevant. During postoperative sedation, the use of ACD-50 significantly reduced CO2 retention. SEV reflection was slightly reduced. However, patients remained sufficiently sedated without increasing SEV infusion.

Background Medical texts such as radiology reports or electronic health records are a powerful source of data for researchers. Anonymization methods must be developed to de-identify documents containing personal information from both patients and medical staff. Although currently there are several anonymization strategies for the English language, they are also language-dependent. Here, we introduce a named entity recognition strategy for Spanish medical texts, translatable to other languages. Results We tested 4 neural networks on our radiology reports dataset, achieving a recall of 97.18% of the identifying entities. Alongside, we developed a randomization algorithm to substitute the detected entities with new ones from the same category, making it virtually impossible to differentiate real data from synthetic data. The three best architectures were tested with the MEDDOCAN challenge dataset of electronic health records as an external test, achieving a recall of 69.18%. Conclusions The strategy proposed, combining named entity recognition tasks with randomization of entities, is suitable for Spanish radiology reports. It does not require a big training corpus, thus it could be easily extended to other languages and medical texts, such as electronic health records.

Background Human tuberculosis (TB) is caused by a plethora of Mycobacterium tuberculosis complex (MTBC) strains belonging to seven phylogenetic branches. Lineages 2, 3 and 4 are considered “modern” branches of the MTBC responsible for the majority of worldwide TB. Since the current BCG vaccine confers variable protection against pulmonary TB, new candidates are investigated. MTBVAC is the unique live attenuated vaccine based on M. tuberculosis in human clinical trials. Methods MTBVAC was originally constructed by unmarked phoP and fadD26 deletions in a clinical isolate belonging to L4. Here we construct new vaccines based on isogenic gene deletions in clinical isolates of the L2 and L3 modern lineages. These three vaccine candidates were characterized at molecular level and also in animal experiments of protection and safety. Findings Safety studies in immunocompromised mice showed that MTBVAC-L2 was less attenuated than BCG Pasteur, while the original MTBVAC was found even more attenuated than BCG and MTBVAC-L3 showed an intermediate phenotype. The three MTBVAC candidates showed similar or superior protection compared to BCG in immunocompetent mice vaccinated with each MTBVAC candidate and challenged with three representative strains of the modern lineages. Interpretation MTBVAC vaccines, based on double phoP and fadD26 deletions, protect against TB independently of the phylogenetic linage used as template strain for their construction. Nevertheless, lineage L4 confers the best safety profile. Funding European Commission (TBVAC2020, H2020-PHC-643381), Spanish Ministry of Science (RTI2018-097625-B-I00), Instituto de Salud Carlos III (PI18/0336), Gobierno de Aragón/Fondo Social Europeo and the French National Research Council (ANR-10-LABX-62-IBEID, ANR-16-CE35-0009, ANR-16-CE15-0003)., Graphical abstract Image, graphical abstract

Civil Guards are deployed throughout Spain and susceptible to being the first responders in out-of-hospital cardiorespiratory arrest. However, their level of training to perform Basic Life Support (BLS) is unknown. The aim of this work was to evaluate the level of knowledge on BLS of Civil Guards, and to explore the association between periodicity of training and knowledge.Cross-sectional study of 839 Civil Guards in Asturias (Spain). A questionnaire with 14 multiple-choice questions assessed knowledge on cardiopulmonary resuscitation (CPR) and use of automatic defibrillator, which were transferred to 0-10 points scales (higher score indicted higher knowledge). Multiple linear regressions were used to estimate mean knowledge scores according to training on BLS (never,2 years ago and ≤2 years ago), adjusted by sociodemographic and occupational variables.Around 1 out of 10 Civil Guards performed some real CPR (11.2%). Regarding training, 42.5% had never participated in courses and 33.4% were trained2 years ago. There was a discordance between willingness to perform BLS (65.6%) and self-perception of preparation (21.8%). A dose-response association was detected between training periodicity and knowledge: 4.26 points (95% CI: 4.07-4.45) of those never formed, 5.93 (95% CI: 5.71-6, 15) of those trained2 years ago, 7.18 (95% CI: 6.92-7.44) of those trained ≤2 years ago (p-trend0.001).The level of training and knowledge on BLS of the Civil Guards is low. Receiving training every two years is significantly associated with greater knowledge.La Guardia Civil es un cuerpo desplegado por toda España y susceptible de ser el primer interviniente en paradas cardiorrespiratorias extrahospitalarias. Sin embargo, se desconoce su nivel de capacitación para realizar Soporte Vital Básico (SVB). El objetivo de este estudio fue estimar el nivel de conocimientos de los guardias civiles sobre SVB y determinar si la periodicidad de la formación se asociaba con los conocimientos.Se realizó un estudio transversal con una muestra de 839 guardias civiles de Asturias. Un cuestionario de 14 preguntas de opción múltiple evaluó los conocimientos sobre resucitación cardiopulmonar (RCP) y uso del desfibrilador automático, que se trasladaron a escalas de 0-10 puntos (más puntos indicaron más conocimientos). Se utilizaron regresiones lineales múltiples para estimar las puntuaciones medias de conocimientos según la formación en SVB (nunca, hace más de 2 años y menor o igual a 2 años), ajustadas por variables sociodemográficas y profesionales.El 11,2% de los guardias civiles había realizado alguna RCP real. El 42,5% nunca había recibido formación y el 33,4% hace más de 2 años. Encontramos discordancia entre la disposición para realizar SVB (65,6%) y la sensación de estar adecuadamente preparado (21,8%). Se detectó asociación dosis-respuesta entre la periodicidad de la formación y los conocimientos: 4,26 puntos (IC95%: 4,07-4,45) de los nunca formados; 5,93 (IC95%: 5,71-6,15) de los formados hace más de 2 años; 7,18 (IC95%: 6,92-7,44) de los formados hace menos o igual a 2 años (p-tendencia0,001).El nivel de formación y conocimientos sobre SVB de los guardias civiles es bajo. Recibir formación cada dos años se asocia significativamente con mayores conocimientos.

Unidad de excelencia María de Maeztu MdM-2015-0552 Altres ajuts: FBBVA research grant (BIOCON_06_106-07) It has been suggested that traditional ecological knowledge (TEK) may play a key role in forest conservation. However, empirical studies assessing to what extent TEK is associated with forest conservation compared with other variables are rare. Furthermore, to our knowledge, the spatial overlap of TEK and forest conservation has not been evaluated at fine scales. In this paper, we address both issues through a case study with Tsimane' Amerindians in the Bolivian Amazon. We sampled 624 households across 59 villages to estimate TEK and used remote sensing data to assess forest conservation. We ran statistical and spatial analyses to evaluate whether TEK was associated and spatially overlapped with forest conservation at the village level. We find that Tsimane' TEK is significantly and positively associated with forest conservation although acculturation variables bear stronger and negative associations with forest conservation. We also find a very significant spatial overlap between levels of Tsimane' TEK and forest conservation. We discuss the potential reasons underpinning our results, which provide insights that may be useful for informing policies in the realms of development, conservation, and climate. We posit that the protection of indigenous cultural systems is vital and urgent to create more effective policies in such realms.

BACKGROUND Hazards of traffic-related air pollution (TRAP) on the developing immune system are poorly understood. We sought to investigate the effects of prenatal exposure to TRAP on cord blood immune cell distributions; and to identify gestational windows of susceptibility. METHODS In-depth immunophenotyping of cord blood leukocyte and lymphocyte subsets was performed by flow cytometry in 190 newborns embedded in the Nutrition in Early Life and Asthma (NELA) birth cohort (2015-2018). Long-term (whole pregnancy and trimesters) and short-term (15-days before delivery) residential exposures to traffic-related nitrogen dioxide (NO2), particulate matter (PM2.5 and PM10), and ozone (O3) were estimated using dispersion/chemical transport modelling. Associations between TRAP concentrations and cord blood immune cell counts were assessed using multivariate Poisson regression models. RESULTS Mean number of natural killer (NK) cells decreased 15% in relation to higher NO2 concentrations (≥36.4 μg/m3) during whole pregnancy (incidence relative risk (IRR), 0.85; 95% CI, 0.72, 0.99), with stronger associations in the first trimester. Higher PM2.5 concentrations (≥13.3 μg/m3) during whole pregnancy associated with a reduced mean number of cytotoxic T cells (IRR, 0.88; 95% CI, 0.78, 0.99). Newborns exposed to higher PM10 (≥23.6 μg/m3) and PM2.5 concentrations during the first and third trimester showed greater mean number of helper T type 1 (Th1) cells (P

The brain uses mainly glucose as fuel with an index of glucose to oxygen utilization close to 6, the maximal index if all glucose was completely oxidized. However, this high oxidative index, contrasts with the metabolic traits of the major cell types in the brain studied in culture, neurons and astrocytes, including the selective use of the malate-aspartate shuttle (MAS) in neurons and the glycerol-phosphate shuttle in astrocytes. Metabolic interactions among these cell types may partly explain the high oxidative index of the brain. In vivo, neuronal activation results in a decrease in the oxygen glucose index, which has been attributed to a stimulation of glycolysis and lactate production in astrocytes in response to glutamate uptake (astrocyte–neuron lactate shuttle, ANLS). Recent findings indicate that this is accompanied with a stimulation of pyruvate formation and astrocyte respiration, indicating that lactate formation is not the only astrocytic response to neuronal activation. ANLS proposes that neurons utilize lactate produced by neighboring astrocytes. Indeed, neurons can use lactate to support an increase in respiration with different workloads, and this depends on the Ca2+ activation of MAS. However, whether this activation operates in the brain, particularly at high stimulation conditions, remains to be established.

Background The ubiquitin-proteasome system (UPS) controls the stability, localization and/or activity of the proteome. However, the identification and characterization of complex individual ubiquitination cascades and their modulators remains a challenge. Here, we report a broadly applicable, multiplexed, miniaturized on-bead technique for real-time monitoring of various ubiquitination-related enzymatic activities. The assay, termed UPS-confocal fluorescence nanoscanning (UPS-CONA), employs a substrate of interest immobilized on a micro-bead and a fluorescently labeled ubiquitin which, upon enzymatic conjugation to the substrate, is quantitatively detected on the bead periphery by confocal imaging. Results UPS-CONA is suitable for studying individual enzymatic activities, including various E1, E2, and HECT-type E3 enzymes, and for monitoring multi-step reactions within ubiquitination cascades in a single experimental compartment. We demonstrate the power of the UPS-CONA technique by simultaneously following ubiquitin transfer from Ube1 through Ube2L3 to E6AP. We applied this multi-step setup to investigate the selectivity of five ubiquitination inhibitors reportedly targeting different classes of ubiquitination enzymes. Using UPS-CONA, we have identified a new activity of a small molecule E2 inhibitor, BAY 11-7082, and of a HECT E3 inhibitor, heclin, towards the Ube1 enzyme. Conclusions As a sensitive, quantitative, flexible, and reagent-efficient method with a straightforward protocol, UPS-CONA constitutes a powerful tool for interrogation of ubiquitination-related enzymatic pathways and their chemical modulators, and is readily scalable for large experiments. Electronic supplementary material The online version of this article (10.1186/s12915-018-0554-z) contains supplementary material, which is available to authorized users.

The insertion Sequence IS6110, only present in the pathogens of the Mycobacterium tuberculosis Complex (MTBC), has been the gold-standard epidemiological marker for TB for more than 25 years, but biological implications of IS6110 transposition during MTBC adaptation to humans remain elusive. By studying 2,236 clinical isolates typed by IS6110-RFLP and covering the MTBC, we remarked a lineage-specific content of IS6110 being higher in modern globally distributed strains. Once observed the IS6110 distribution in the MTBC, we selected representative isolates and found a correlation between the normalized expression of IS6110 and its abundance in MTBC chromosomes. We also studied the molecular regulation of IS6110 transposition and we found a synergistic action of two post-transcriptional mechanisms: a -1 ribosomal frameshift and a RNA pseudoknot which interferes translation. The construction of a transcriptionally active transposase resulted in 20-fold increase of the transposition frequency. Finally, we examined transposition in M. bovis and M. tuberculosis during laboratory starvation and in a mouse infection model of TB. Our results shown a higher transposition in M. tuberculosis, that preferably happens during TB infection in mice and after one year of laboratory culture, suggesting that IS6110 transposition is dynamically adapted to the host and to adverse growth conditions., Author summary Since the pioneering discovery of transposition by Barbara McClintock in eukaryotes and later in prokaryotes by Robert W. Hedges and Alan E. Jacob, it has become clear the key role of mobile genetics elements in chromosome remodelling, microbial evolution and host adaptation. The insertion sequence IS6110 is widely recognized for its utility in TB diagnosis and epidemiology because it is only present in the M. tuberculosis Complex (MTBC) and its transposition provides an excellent chromosomal polymorphic variability allowing the study of recent TB transmission. This inherent feature of IS6110 leads us to hypothesize that IS6110 plays a crucial role during the TB infectious cycle. However, the biological significance of IS6110 has been hindered by its almost exclusive use as an epidemiological marker. Here, we study the regulatory mechanisms and the distribution of IS6110 in the different MTBC lineages. We discuss the potential biological implications of IS6110, that is much more than an excellent TB epidemiological tool. Since IS6110 could play an important role in the adaptation of MTBC to the host, this study opens new avenues to decipher the biological roles of IS6110 in TB pathogenesis.

The aim of the present study was to evaluate the efficacy and safety of non-supervised home-based exercise versus individualized and supervised programs delivered in clinic-based settings for the functional recovery immediately after discharge from a primary TKA. Medline, Embase, Cochrane, and PEDro databases were screened, from inception to April 2015, in search for randomized clinical trials (RCT) of home-based exercise interventions versus individualized and supervised outpatient physical therapy after primary TKA. Target outcomes were: knee range of motion (ROM), patient-reported pain and function, functional performance, and safety. Risk of bias was assessed with the PEDro scale. After assessing homogeneity, data were combined using random effects meta-analysis and reported as standardized mean differences or mean differences. We set a non-inferiority margin of four points in mean differences. The search and selection process identified 11 RCT of moderate quality and small sample sizes. ROM active extension data suitable for meta-analysis was available from seven studies with 707 patients, and ROM active flexion from nine studies with 983 patients. Most studies showed no difference between groups. Pooled differences were within the non-inferiority margin. Most meta-analyses showed significant statistical heterogeneity. Short-term improvements in physical function and knee ROM do not clearly differ between outpatient physiotherapy and home-based exercise regimes in patients after primary TKA; however, this conclusion is based on a meta-analysis with high heterogeneity. I.

DOI: 10.1016/j.ctcp.2016.08.005URL: http://www.sciencedirect.com/science/article/pii/S1744388116300706Filiació URV: SIMemòria Recent studies on reflexology describe the appearance of different application-associated effects, attributed to a self-regulatory mechanism related to treatment efficacy. On the other hand, sleep is a physiological process of vital importance for health. Its main value lies in restoring the natural balance between neuronal centers. Among its associated behavioral characteristics are spontaneous movements and eye movements. The aim of this study is to investigate the effects that occur during application of reflexology and that are not described in the literature. This is a descriptive observational study with a quantitative methodology. Abivariate anlysis has been conductec through chi-square test or Anova as apropiate. A total of 111 clients of a therapy center in Tarragona have participated in the study. They were assigned into four groups (musculoskeletal, stress, anxiety, mantenance). Reflexology was administered and observered the manifestations that occured during the session. The findings have identified four categories of effects, of which there was no previous reference. These effects can be related to any of the stages of sleep. This study shows that reflexology promotes its application for different effects, such as eye movements and spontaneous movements. These data reveal the need to investigate these effects and their impact on health as well as their possible relationship with sleep.

espanolAntecedentes: los pacientes criticos suelen desarrollar un estado de estres catabolico que se traduce en una respuesta infl amatoria sistemica (RIS) que altera los biomarcadores clinico-nutricionales, aumentando las necesidades de nutrientes y de energia. Objetivo: evaluar el estatus de albumina, prealbumina y transferrina en pacientes criticamente enfermos y la asociacion entre estos parametros clinico-nutricionales con la severidad del paciente durante siete dias de estancia en la unidad de cuidados intensivos (UCI). Metodo: estudio multicentrico, de seguimiento, prospectivo, observacional y analitico. Un total de 115 sujetos en estado critico fueron incluidos en el estudio. Los parametros clinico-nutricionales y la gravedad clinica fueron controlados al ingreso y al septimo dia de estancia en UCI. Resultados: una disminucion signifi cativa en la gravedad del paciente (p Conclusion: los resultados del presente estudio apoyan la idea de incluir la prealbumina como predictor de la gravedad dentro de la escala APACHE II debido a la asociacion encontrada entre la gravedad y un estatus pobre de prealbumina. EnglishIntroduction: Critically ill patients typically develop a catabolic stress state as a result of a systemic inflammatory response (SIRS) that alters clinical-nutritional biomarkers, increasing energy demands and nutritional requirements. Objective: To evaluate the status of albumin, prealbumin and transferrin in critically ill patients and the association between these clinical-nutritional parameters with the severity during a seven day stay in intensive care unit (ICU). Method: Multicenter, prospective, observational and analytical follow-up study. A total of 115 subjects in critical condition were included in this study. Clinical and nutritional parameters and severity were monitored at admission and at the seventh day of the ICU stay. Results: A significant decrease in APACHE II and SOFA (p Conclusion: The results of the present study support the idea of including low prealbumin status as a severity predictor in APACHE II scale, due to the association found between severity and poor status of prealbumin.

Anthracofibrosis is a bronchial stenosis due to local mucosal fibrosis that also presents anthracotic pigment in the mucosa. The cause has not been well clarified, although there is a frequent association with tuberculosis and the exposure to smoke from biofuel or biomass combustion. It is an entity that has not been reported in Spain, although the influx of people from rural areas of developing countries or rural areas of our own country should make us contemplate this entity in the differential diagnosis of our patients. We present 3 cases detected in Spain (2 of them natives) diagnosed by bronchoscopy and bronchial biopsy, which are techniques necessary to confirm the diagnosis. There is no specific treatment, except for tuberculostatic treatment in cases with coexisting tuberculosis.

trauma and severe infections cause remarkable metabolic changes in patient with SIRS from an adaptive response aimed to control the underlying disease, repairing damaged tissue, and to synthesize substrates. If the attack is intense and sustained and the patient has a compromised nutritional status, can evolve into multiple organ failure and death.assessment of nutritional proteic status and the involvement of proteins and inflammatory factors in critically ill patients.multicenter observational analytical study in critical ill patients at the admission in ICU.patients showed disturbances in clinical nutritional parameters which confirm their hypercatabolic situation, showing malnutrition state at admission, where 42.9% had plasma levels below the reference prealbumin. Amino acid profile was situated below the reference values and 99% of patients had low plasma transferrin. Significant differences were observed in total protein, ferritin and transferrin parameters adjusted by CRP levels, being higher when patients presented high inflammation in the case of ferritin and the opposite for the rest of parameters. Adjusting APACHE and SOFA scores according to low, medium and high severity, results showed significant differences in creatinine, urea, and transferrin, being lower at high severity grade for the last one.critical illness is characterized by a high degree of stress and accelerated degradation of proteins that cause malnutrition, systemic inflammation and organ dysfunction, with a significant association between albumin, ferritin and transferrin.Antecedentes: el trauma y las infecciones severas causan cambios metabólicos notables en los pacientes con SRIS como una respuesta adaptativa dirigida a controlar la enfermedad subyacente, la reparación del tejido dañado y para sintetizar sustratos. Si el ataque es intenso y sostenido y el paciente tiene un estado nutricional comprometido puede evolucionar a insuficiencia orgánica múltiple y muerte. Objetivo: evaluación del estado nutricional proteico y la participación de las proteínas y los factores inflamatorios en pacientes críticamente enfermos. Método: estudio analítico observacional multicéntrico en pacientes enfermos críticos en la admisión en la UCI. Resultados y discusión: los pacientes mostraron alteraciones en los parámetros nutricionales clínicos que confirman su situación hipercatabólica, mostrando malnutrición a la admisión en UCI, donde el 42,9% tenían niveles plasmáticos de prealbúmina por debajo de la referencia. Los aminoácidos se encuentran por debajo de los valores de referencia y el 99% de los pacientes presentaron bajos niveles plasmáticos de transferrina. Se observaron diferencias significativas en los niveles de proteína total, ferritina y transferrina ajustados por los niveles de PCR, siendo mayor cuando los pacientes presentaron altos valores de inflamación, en el caso de la ferritina, y lo opuesto para el resto de parámetros. Al estratificar por las puntuaciones APACHE y SOFA de acuerdo a la gravedad baja, media y alta, los resultados mostraron diferencias significativas en creatinina, urea y transferrina, siendo menor cuanto mayor era el grado de severidad para la transferrina. Conclusión: la enfermedad crítica se caracteriza por un alto grado de estrés y la degradación acelerada de proteínas que causan malnutrición, inflamación sistémica y la disfunción de órganos, con una asociación significativa entre albúmina, ferritina y transferrina.

Rotavirus (RV) epidemiology presents differences between developing and developed countries among which are seasonality age at first infection variability of strain in circulation and severity of disease. Since in Venezuela we have distinct seasonal patterns of RV occurrence we examined the epidemiological profile of RV disease associated to these differences by analyzing data from previous studies conducted in Venezuela. Data were collected from children

The immune response elicited by the rotavirus nonstructural protein NSP4 and its potential role in protection against rotavirus disease are not well understood. We investigated the serological response to NSP4 and its correlation with disease protection in sera from 110 children suffering acute diarrhea, associated or not with rotavirus, and from 26 children who were recipients of the rhesus rotavirus tetravalent (RRV-TV) vaccine. We used, as antigens in an enzyme-linked immunosorbent assay (ELISA), affinity-purified recombinant NSP4 (residues 85 to 175) from strains SA11, Wa, and RRV (genotypes A, B, and C, respectively) fused to glutathione S -transferase. Seroconversion to NSP4 was observed in 54% (42/78) of the children who suffered from natural rotavirus infection and in 8% (2/26) of the RRV-TV vaccine recipients. Our findings indicate that NSP4 evokes significantly ( P < 0.05) higher seroconversion rates after natural infection than after RRV-TV vaccination. The serum antibody levels to NSP4 were modest (titers of ≤200) in most of the infected and vaccinated children. A heterotypic NSP4 response was detected in 48% of the naturally rotavirus-infected children with a detectable response to NSP4. Following natural infection or RRV-TV vaccination, NSP4 was significantly less immunogenic than the VP6 protein when these responses were independently measured by ELISA. A significant ( P < 0.05) proportion of children who did not develop diarrhea associated with rotavirus had antibodies to NSP4 in acute-phase serum, suggesting that serum antibodies against NSP4 might correlate with protection from rotavirus diarrhea. In addition, previous exposures to rotavirus did not affect the NSP4 seroconversion rate.

Background: A live attenuated monovalent rotavirus vaccine RIX4414 was developed with a human strain of G1P1A P[8] specificity to reduce the rotavirus burden in children. Methods: A double blind, randomized, placebo-controlled study evaluated the efficacy, immunogenicity, safety and reactogenicity of 2 oral doses of RIX4414 (10 4.7 , 10 5.2 or 10 5.8 focus-forming units) at 2 and 4 months coadministered with routine vaccinations and oral poliovirus vaccine given for study purposes at least 14 days apart. The 2155 infants (1618 vaccine/537 placebo) enrolled in Brazil, Mexico and Venezuela were followed until 1 year of age. Results: Antirotavirus IgA seroconversion rates 2 months after dose 2 ranged between 61% (10 4.7 ffu group) and 65% (10 5.8 ffu group), and most of the infants had seroprotective levels of antibodies to coadministered routine vaccinations. The reactogenicity profile of RIX4414 was similar to that of the placebo, and no vaccination-related serious adverse events were reported. Protective efficacy against severe and any rotavirus gastroenteritis from 15 days post-dose 2 was highest in the 10 5.8 ffu group [86%; 95% confidence interval (95% CI), 63-96% and 70% (95% CI 46-84%), P < 0.001, 2-sided Fisher's exact test]. The efficacy against hospitalization was 79% (95% CI 48-92%) for pooled vaccine groups. Multiple rotavirus serotypes [Gl (50%), G9 (40%), G2, G3 and G4] were identified from gastroenteritis stools (enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction) during the study period. For severe gastroenteritis caused by G9 serotypes, the protection reached 77% (95% CI 18-96%) in the 10 5.8 ffu group, providing proof of concept that the monovalent G1P1A P[8] human rotavirus vaccine elicits cross-protection against the G9 strain. A reduction in any and severe rotavirus gastroenteritis was already observed at post-dose 1 (period: day of dose 1 to 14 days post-dose 2) in vaccinees compared with placebo recipients. Conclusions: Two doses of RIX4414 are highly efficacious, providing cross-protection (Gl and G9 strains, prevalent during this study) and early protection against any and severe rotavirus gastroenteritis and hospitalization to infants in Latin America.

To effectively introduce a rotavirus vaccine in developing countries it is necessary to estimate the burden of rotavirus disease. Therefore we examined the epidemiologic and clinical features of rotavirus diarrhea in children younger than 5 years of age in Carabobo State and extrapolated these results to the rest of Venezuela. From January 1 1998 to December 31 2002 we conducted surveillance for rotavirus diarrhea in all children younger than 5 years of age at the Ciudad Hospitalaria “Dr Enrique Tejera” in Valencia (Carabobo). Rotavirus is the major cause of diarrhea in children younger than 5 years of age in Venezuela accounting for one-fourth (23%) of all episodes of diarrhea requiring medical treatment and one-third (33%) of those requiring hospitalization. Rotavirus diarrhea was responsible for 3% of all hospitalizations and 2% of all medical visits. In Valencia rotavirus had a marked seasonal peak during the dry and cold months of the year. Rotavirus was most frequent in children 3–23 months of age and 61% of the cases occurred by the age of 1 year. In addition rotavirus diarrhea was more severe in younger children. Overall by the age of 5 years 1 child in 72 will be hospitalized and 1 in 24 will visit the clinic for rotavirus disease. In Venezuela we estimated that each year 118000 children experience rotavirus illness requiring medical care and 39000 children require hospitalization. This study confirms the significant impact of rotavirus disease in Venezuela. Rotavirus vaccines currently in development could diminish the morbidity associated with this common cause of childhood diarrheal disease. (authors)

Background. The first licensed rotavirus vaccine was withdrawn from use in the United States because of a low risk of intussusception. Consequently tests of new rotavirus vaccines will require some baseline knowledge of the rates and treatment of intussusception in countries where these vaccines will be tested. Therefore the objective of this study was to assess hospitalization rates and describe the epidemiologic and clinical characteristics of intussusception in Carabobo, Venezuela. Methods. This study reviewed hospital data and clinical records of pediatric patients with intussusception admitted to eight hospitals in Carabobo between January 1, 1998 and December 31, 2001. Results. For the 4-year period the average annual hospitalization rate for intussusception among infants (

During a phase 3 clinical trial of rhesus monkey rotavirus-based quadrivalent vaccine in Venezuela, 2207 infants received 3 oral doses of vaccine (4 x 105 plaque-forming units/dose) or placebo at ages approximately 2, 3, and 4 months; 219 (14%) of 1537 stools obtained during 1550 diarrheal episodes in postvaccination surveillance were rotavirus-positive by enzyme-linked immunosorbent assay. With the use of various VP7 and VP4 primers for genotyping purposes, 213 of 219 rotavirus-positive stools were analyzed by reverse-transcription polymerase chain reaction. Twenty-nine (14%) of 213 rotavirus-positive stools contained at least 2 distinct rotavirus strains: a low-titered vaccine strain(s) and a second strain that, when possible, was studied further and found to be a wild-type rotavirus strain. The titer of vaccine viruses in 19 stools that plaqued directly in cell cultures ranged from 10(1) to 10(3) plaque-forming units/0.5 mL of a 10% stool suspension. Reassortants of vaccine virus and wild-type human rotavirus were not detected.

BACKGROUND Knowledge of the impact of rotavirus-associated disease on the health care systems of South America can aid in defining strategies for diagnosis, management and prevention. Up to date information on the impact of rotavirus disease in South America is scarce. AIM To determine prospectively the impact of rotavirus disease as a cause of medical visits and hospitalizations at three large sentinel pediatric hospitals in Argentina, Chile and Venezuela. METHODS A 2-year prospective surveillance for rotavirus-associated medical visits and hospitalizations was conducted during 1997 through 1998 at three large sentinel public hospitals, one each in Argentina, Chile and Venezuela. A common surveillance protocol was implemented at the three sites, and a representative number of nonbloody diarrhea stool samples from children

Rotaviruses are the principal known etiologic agents of severe diarrhea among infants and young children worldwide. Although a rhesus rotavirus-based quadrivalent vaccine is highly effective in preventing severe diarrhea in developed countries, in developing countries its efficacy has been less impressive. We thus conducted a catchment study in Venezuela to assess the efficacy of the vaccine against dehydrating diarrhea.In this randomized, double-blind, placebo-controlled trial, 2207 infants received three oral doses of the quadrivalent rotavirus vaccine (4x10(5) plaque-forming units per dose) or placebo at about two, three, and four months of age. During approximately 19 to 20 months of passive surveillance, episodes of gastroenteritis were evaluated at the hospital.The vaccine was safe, although 15 percent of the vaccinated infants had febrile episodes (rectal temperature,or =38.1 degrees C) during the six days after the first dose, as compared with 7 percent of the controls (P0.001). However, the vaccine gave 88 percent protection against severe diarrhea caused by rotavirus and 75 percent protection against dehydration, and produced a 70 percent reduction in hospital admissions. Overall, the efficacy of the vaccine against a first episode of rotavirus diarrhea was 48 percent. Horizontal transmission of vaccine virus was demonstrated in 15 percent of the vaccine recipients and 13 percent of the placebo recipients with rotavirus-positive diarrhea.In this study in a developing country, the quadrivalent rhesus rotavirus-based vaccine induced a high level of protection against severe diarrheal illness caused by rotavirus.

In this study, age at menopause was examined in relation to demographic and life style factors among Latin-American immigrants to Madrid and their Spanish counterparts.Respondents were drawn from the Decisions at Menopause Study (2002-2003) and from a recent sample of Latin-American immigrants to Madrid (2010-2011). The final sample included 484 women after excluding women with induced menopause and use of HT. Probit analyses and Cox proportional hazard models were used to estimate median age at menopause and to identify factors associated with an early age at menopause.Median estimated age at natural menopause was 52.0 years (51.2-53.0) for Spanish women and 50.5 years (49.9-51.2) for Latin-American women. Immigrant women were more likely to reach menopause at an earlier age after controlling for confounding factors. Nulliparity and lower levels of education were associated with an earlier age at menopause. A higher body mass index was associated with a later age at menopause in the Spanish model. Among the Latin-American sample, women from the Dominican Republic and women who underwent menopause before migrating were more likely to reach menopause at an earlier age.The results reported here demonstrate that early life events, including place of birth, and later life events, such as timing of migration, were associated with age at menopause. This study highlights the importance of taking into account differences in the age of onset of menopause in the multicultural population of Madrid when considering the health of women at midlife and beyond.

The most extensively explored strategy for rotavirus vaccination has been the Jennerian approach, which uses an antigenically related rotavirus strain from an animal host as the immunogen to induce protection against the 4 epidemiologically important group A rotavirus VP7 serotypes. Because this approach has shown limited efficacy, a modified Jennerian approach was developed with the goal of achieving broader antigenic coverage. Four VP7 serotypes were incorporated into a quadrivalent vaccine comprised of three rhesus-human rotavirus reassortants, each with 10 rhesus rotavirus genes and 1 human rotavirus gene that encodes VP7 serotype 1, 2, or 4 specificity ; the rhesus rotavirus itself provides coverage for VP7 serotype 3. This approach appears quite promising for preventing severe rotavirus diarrhea, including those episodes that lead to dehydration. Additional strategies under development stress the role not only of human rotavirus VP7 but also of human rotavirus VP4, the other outer capsid protein that also induces neutralizing antibodies.

In response to stress or injury the heart undergoes a pathological remodeling process, associated with hypertrophy, cardiomyocyte death and fibrosis, that ultimately causes cardiac dysfunction and heart failure. It has become increasingly clear that signaling events associated with these pathological cardiac remodeling events are regulated by scaffolding and anchoring proteins, which allow coordination of pathological signals in space and time. A-kinase anchoring proteins (AKAPs) constitute a family of functionally related proteins that organize multiprotein signaling complexes that tether the cAMP-dependent protein kinase (PKA) as well as other signaling enzymes to ensure integration and processing of multiple signaling pathways. This review will discuss the role of AKAPs in the cardiac response to stress. Particular emphasis will be given to the adaptative process associated with cardiac hypoxia as well as the remodeling events linked to cardiac hypertrophy and heart failure. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Cardiac Pathways of Differentiation, Metabolism and Contraction.

Olfactory ensheathing cells are glial cells located in the olfactory bulb and nerve. Microscopically, both olfactory ensheathing cells and Schwann cells have similar morphological and immunohistochemical features. However, olfactory ensheathing cells are negative for Leu-7(CD-57), whereas Schwann cells are positive. We present the case of a 49 year-old male with a history of visual impairment and hyposmia. Radiological CT and MRI studies showed a subfrontal cystic extra-axial mass, which eroded the right cribriform plate, with heterogeneous contrast enhancement. Total excision of the tumour was performed by bifrontal craniotomy. Histological examination initially suggested a schwannoma, with immunohistochemical staining being positive for S-100 protein and negative for epithelial membrane antigen (EMA). However, the tumour was negative for Leu-7. Accordingly, the final diagnosis was olfactory ensheathing cell tumour. Herein, we describe the sixth case of intracranial olfactory ensheathing cell tumour and stress the important role of immunohistochemical techniques in obtaining a definitive diagnosis.

We evaluated the reactogenicity and antigenicity of a quadrivalent rotavirus vaccine composed of serotype 3 rhesus rotavirus (RRV) and three single-gene-substitution reassortants of RRV and human strain D (D x RRV, serotype 1), DS1 (DS1 x RRV, serotype 2), or ST3 (ST3 x RRV, serotype 4) in a double-masked study with 302 infants in Caracas, Venezuela. Three doses of the quadrivalent vaccine composed of either 10(5) PFU (low titer) or 10(6) PFU (high titer) of each component were administered to 99 and 101 infants, respectively, at 4-week intervals starting at the second month of age; 102 infants received a placebo. Postvaccination reactions were monitored by home visits every other day during the week postvaccination. The vaccine was associated with the occurrence of mild, short-lived febrile episodes in 26 and 23% of the recipients after the first doses of high- or low-titer vaccine, respectively, in comparison with 13% of the infants receiving the placebo. Febrile reactions occurred less frequently in vaccinees after the second or third dose than after the initial dose. The vaccine was not significantly associated with diarrhea or any additional symptom or sign. Serum specimens obtained shortly before the first, 4 weeks after the first, and 4 weeks after the third dose of vaccine or placebo were tested by an immunoglobulin A enzyme-linked immunosorbent assay and by neutralization assays. Seroresponses occurred significantly more often after 3 doses than after a single dose of either vaccine. Immunoglobulin A responses were observed in 80 and 79% of the infants after 3 doses of high- or low-titer vaccine, respectively. Most of the infants tested developed a neutralization response to RRV after 3 doses of the high- (90%) or low-(88%) titer vaccine. Neutralization response rates to human rotavirus serotypes 1 to 4 after 3 doses were similar in both vaccine and 87 of 90 receiving the high-titer vaccine developed seroresponses, as detected by any of the assays employed. The study indicates that 3 doses of quadrivalent vaccine at a titer of 10(6) PFU of each component offered no advantage over the lower-titer preparation for use in efficacy trials.

To have a better knowledge of the epidemiological and clinical aspects of rotavirus (RV) infection treated at out-patient clinics, we carried out a study in five centers (one hospital type I, and four out-patient clinics) of Miranda state, between february 2006 and january 2007. We evaluated 194 children5 years old with acute diarrhea and analyzed the following characteristics: gender, age, socioeconomic condition, breastfeeding, nutritional status, clinical characteristics, rotavirus diagnosis and typing by Enzyme Linked immuno Sorbent Asssay (EISA) and G typing by RT-PCR, respectively. The population presented the following characteristics: 53% males; mean age, 17.6 +/- 14.05 months; 68% belonged to worker and marginal class (Graffar 4 and 5); 59% were breast-fed during their first year of life; 13% were malnourished; 40% were dehydrated and 19% were infected with RV. RV infection was significantly and more frequently associated with vomiting (94%, P0.0001) and malnourishment (30%, P=0.0010) than the episodes negative for rotavirus. In dehydrated cases, significantly greater malnourishment (21%, P=0.0232) and RV (28%, P=0.0407). We found significant differences between the hospital and out-patient clinics, in terms of Graffar 4 and 5 (90% vs .53%, P0.0001), malnourished (23% vs. 6%, P=0.006), dehydration (63% vs 24%, P0.0001) and RV (27% vs. 13%, P=0.027). Fourteen samples RV positive were genotyped and the G types were 29% G1, 21% G3 and 14% G4 types. This study suggests that RV cause severe diarrhea associated to malnutrition and poverty and significantly affect the population older than one year of age in the region studied.

Phase I studies of an oral quadrivalent rotavirus vaccine were conducted in 130 Venezuelan infants 10 to 20 weeks of age. The vaccine consists of a mixture of equal amounts of rhesus rotavirus (RRV) vaccine (serotype 3 [VP7]) and each of three human rotavirus-RRV reassortant strains: D x RRV (serotype 1 [VP7]), DS1 x RRV (serotype 2 [VP7]), and ST3 x RRV (serotype 4 [VP7]). Three different doses of the quadrivalent vaccine (0.25 x 10(4), 0.5 x 10(4), and 10(4) PFU of each component) were evaluated sequentially for safety and antigenicity in placebo-controlled, double-blind trials. Starting the day after vaccination, the infants were monitored by daily home visits for 7 days. Only minor reactions were observed during this period; these were limited to mild transient febrile episodes which began day 2 or 3 after vaccination and lasted 1 to 2 days in 15 to 30% of the infants. Serological studies demonstrated that 68 to 96% of the infants developed a rotavirus serum immunoglobulin A response following vaccination. However, when tested by plaque reduction neutralization assay against individual human rotavirus serotype 1, 2, 3, or 4, the response rates ranged from 4 to 23% with the low dose, 21 to 33% with the medium dose, and 32 to 58% with the high dose. Most (73 to 79%) infants developed neutralizing antibodies to RRV following administration of each dose schedule. Vaccine virus shedding was analyzed by utilizing tissue culture isolation of virus from stool. All of the infants who received the lower of medium dose and 89% of those fed the high dose shed one or more components of the vaccine. Analyses of rotavirus serotypes isolated from the stool of infants who received the 0.25 x 10(4) -PFU dose revealed that DS1 x RRV was the most commonly shed vaccine component, followed by RRV, D x RRV, and ST3 x RRV in that order.

The efficacy of a rhesus rotavirus vaccine (MMU 18006, serotype 3) against infantile diarrhea was evaluated by active home surveillance of a group of 320 children 1-10 months of age in Caracas, Venezuela. During a 1 year period following oral administration of vaccine or placebo under a double-masked code, over 600 diarrheal episodes were detected. Etiologic studies revealed that heat-stable toxin (ST) producing enterotoxigenic E. coli (ETEC) was the most common diarrheal agent detected (34%) followed by enteropathogenic E. coli (EPEC, 10.9%), heat-labile toxin (LT) producing ETEC (7.6%), rotavirus (6.9%), Cryptosporidium (4.8%) and Campylobacter (1.3%). ST-producing ETEC were also recovered from over 20% of control stool specimens obtained during diarrhea-free periods, whereas EPEC, rotavirus, Cryptosporidium, and Campylobacter were rarely detected in such control specimens. Rotavirus was responsible for about one-half of the more severe cases of diarrhea. Twenty-two of 151 infants who received placebo (14.6%) and eight of 151 receiving a 10(4) PFU dose of vaccine (5.3%) had rotavirus diarrhea during the follow-up period for an efficacy level of 64% against any rotavirus diarrhea. However, vaccine efficacy reached 90% against the more severe cases of rotavirus diarrhea and was noticeably high in the 1-4 month age group. Serotypic analysis of the rotaviruses detected suggests that the resistance induced by the vaccine was type specific since significant protection was only evident against serotype 3 rotaviruses. A 10(3) PFU dose tested initially in 18 children did not appear to protect against rotavirus diarrhea.

The feces of 397 patients with acute diarrheal disease (ADD) and of other 121 patients without diarrea (control group) were studied in the state of Mérida, Venezuela, from June 1993 to December 1994. The genus Aeromonas was identified in patients with ADD in 11.83% and in 5.78% of the patients from the control group. On studying the virulence factors described for Aeromonas (enterotoxin, cytotoxin, hemaglutinins, cellular hydrofibrosity, and hemolytic activity) in the isolated strains, it was detected that all presented at least one of the factors investigated associated with enteropathogenicity. Of the isolated species, Aeromonas caviae was the most frequently identified. All these results suggest that the Aeromonas species are potential enteric pathogens in this population.

Hospital-based studies to determine the etiology of deaths from diarrhea are scarce. In this study we specifically analyzed deaths due to rotavirus to assess the rotavirus impact on diarrhea mortality. To determine the rotavirus proportion contributing to mortality due to diarrhea we analyzed data obtained from a hospital-based mortality surveillance conducted over 7 years in the Ciudad Hospitalaria Dr. Enrique Tejera Valencia Venezuela. Rotavirus was identified in stool samples collected from children who died of diarrhea by a confirmatory ELISA and/or reverse transcription polymerase chain reaction. Our results show that rotavirus (21%; 21/100) is the leading cause of death due to diarrhea among children < 5 years of age; rotavirus also has an important impact (2%; 21/1336) on deaths from all causes in this age group. Shigella spp. (19%; 13/69) was the second most important cause of death followed by calicivirus (6%; 3/53). Furthermore this study documents a seasonal pattern in the deaths due to rotavirus (odds ratio 3.28; 95% confidence interval 1.13-9.76). For Venezuela it is estimated that approximately 300 children < 5 years of age die of rotavirus each year which means that 1 in 1800 children die by the age of 5. Rotavirus was found to be the main cause of death due to diarrhea which supports previous estimations. This is the first study to present data of cause-specific mortality due to diarrhea based on hospital surveillance of diarrhea etiologies. (authors)

To assess the cost of medical care for rotavirus gastroenteritis and the cost-effectiveness of the antiretroviral vaccine in Venezuelan children under five.We used an economic model that comprises epidemiologic information, vaccine efficacy, and the cost of medical care in connection with rotavirus gastroenteritis, viewed from a social perspective. In order to determine the effectiveness of the vaccine, we estimated the number of hospitalized cases, of medical visits, and of deaths averted after vaccination. The cost-effectiveness of the vaccine was determined on the basis of the number of disability-adjusted life years (DALYs) and cases averted.In Venezuela, health services spend approximately 4.2 million US$ yearly on covering the costs of medical care for rotavirus-related disease. In a vaccinated cohort, an antiretroviral vaccination program would prevent around 52% (186) of the deaths, 54% (7,232) of the hospitalizations, and 50% (55,168) of the ambulatory visits that take place during the first five years of life. For an estimated cost of approximately 24 US$ per individual vaccination schedule, the cost-effectiveness ratio obtained is 1,352 US$ per DALY.The results of this study suggest that antiretroviral vaccination is a cost-effective strategy for preventing rotavirus gastroenteritis in Venezuela, since it can prevent deaths and DALYs in the population under five years of age.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas, Bel?m, PA, Brasil. Instituto Nacional de La Nutricion. Mexico City, Mexico. Instituto Nacional de La Nutricion. Mexico City, Mexico. Instituto de Biomedicina, Caracas, Venezuela. Instituto de Biomedicina. Caracas, Venezuela. GlaxoSmithKline Biologicals Latin America & Caribbean. Belgium. GlaxoSmithKline Biologicals Latin America & Caribbean. Belgium. GlaxoSmithKline Biologicals Latin America & Caribbean. Belgium. GlaxoSmithKline Biologicals Latin America & Caribbean. Belgium. GlaxoSmithKline Biologicals Latin America & Caribbean. Belgium. GlaxoSmithKline Biologicals. Belgium GlaxoSmithKline Biologicals. Belgium. An oral, human-derived monovalent (G1P1A) rotavirus vaccine, strain RIX4414, has been developed by GlaxoSmithKline, Rixensart, Belgium. The safety, immunogenicity and efficacy of this vaccine were evaluated in a randomized, double-blind, placebo-controlled, phase IIb trial conducted in Brazil, Mexico and Venezuela. Healthy infants were given two doses of vaccine (104.7, 105.2 or 105.8 ffu) or placebo at age 2 and 4 months, with routine DTPw-HBV and Hib vaccines. OPV was given separately, at least 2 weeks before or after administration of the study vaccine. A total of 2155 infants were enrolled, of whom 1618 received one of the three vaccine viral concentrations and 537 were given placebo. Analysis of efficacy included diarrheal episodes occurring from 2 weeks after second dose until one year of age. Efficacy rates against any rotavirus gastroenteritis, severe rotavirus gastroenteritis and hospitalizations for rotavirus disease were as high as 70% (46?84%; 95%CI), 86% (63?96%; 95%CI), and 93% (54?100%; 95%CI), respectively. For non-G1 (mainly G9) serotypes, RIX4414 vaccine conferred protection as high as 83% (40?97%; 95%CI) against severe gastroenteritis. A decrease was noted in the incidence of severe rotavirus-related gastroenteritis after first dose. It is demonstrated that two doses of RIX4414 are highly efficacious against severe rotavirus gastroenteritis and hospitalization, including disease caused by non-G1 strains, namely G9 serotypes.

To directly compare serum rotavirus specific IgA as a marker of protection in children vaccinated with the RRV-TV (Rotashield) vaccine and in naturally infected children, we studied pre-existing rotavirus IgA antibodies by ELISA assays in these groups of children within the first 5 days after the onset of a diarrhea episode, due or not to rotavirus. In immunized children, rotavirus IgA titers were similar between infected and non-RV infected children. In non-immunized children, the proportion with rotavirus IgA titers was significantly greater in non-RV infected children (58%) than in infected children (31%). Additionally, a titer >/=1:800 was associated with 68% protection. Thus, in this study serum rotavirus IgA showed a good correlation with protection in children pre-exposed to natural infection but not in those immunized with the RRV-TV vaccine.

The need for safe and effective vaccines to reduce morbidity and mortality caused by rotavirus gastroenteritis in children is well-known. A live attenuated monovalent rotavirus vaccine (Rotarix) containing human rotavirus strain RIX4414 of G1P1A P[8] specificity is being developed to meet the global need. An overview of RIX4414 trials in developed and developing settings is presented for 3 selected trials conducted in Finland (pilot study), Latin America (Brazil, Mexico and Venezuela) and Singapore involving 5024 infants. The vaccine was well-tolerated, with no increase in any solicited symptoms as compared with the placebo. After 2 doses, 61-91% of vaccinated infants developed rotavirus-specific IgA antibodies. There was no interference with immunogenicity of coadministered routine pediatric vaccines. Rotarix significantly reduced rotavirus gastroenteritis episodes and rotavirus-related hospitalizations in vaccinated infants compared with placebo recipients (P < 0.05). Vaccine efficacy was observed against severe rotavirus gastroenteritis caused by G1 and non-G1 types including the emerging G9 type (P < 0.05) in Latin America. These results show prospects for widespread use of Rotarix to reduce rotavirus disease burden and warrant continued worldwide evaluation.

This report describes the results of additional analyses of the trial carried out with the rhesus rotavirus-based quadrivalent vaccine in Venezuela. In the present study, we re-examined the data from this previous rotavirus vaccine trial to assess the statistical interaction between vaccine efficacy and (i) the duration of efficacy into the second year of life, (ii) socioeconomic conditions, and (iii) rotavirus seasonality. We found that among Venezuelan children, the rotavirus vaccine confers protection against severe diarrhea during the first 2 years of life independently of socioeconomic conditions and seasonality.

The epidemiological features of rotavirus infection may be quite relevant for evaluation of the performance of a rotavirus vaccine in different settings, as well as for monitoring its impact during vaccination under routine conditions. This article describes some important issues regarding rotavirus epidemiology in Venezuela, where major field trials of rotavirus vaccine have been carried out. Rotaviruses was significantly more frequently observed in inpatient (43%) than in outpatient (21%) consultations for diarrhea in infants and young children. There was a high prevalence of rotavirus illness, regardless of socioeconomic conditions, but the risk of dehydration was greater among the lower socioeconomic groups. Rotavirus disease occurs year-round, with a slight seasonal pattern. Eighty-five percent of rotavirus-positive diarrheal episodes, as well as 86% of cases of dehydration due to rotavirus, occurred during the first year of life. However, rotavirus illnesses occur less commonly during the first months of life (0-2 months), which may be a result of protection by transplacental antibodies. The pattern of acquisition of rotavirus antibody was consistent with this age distribution of disease and with optimal age for vaccination. Thus, regional epidemiological characteristics of rotavirus infection may affect optimal performance of rotavirus vaccine.

To evaluate the epidemiological significance of HEp-2 cell-adherent Escherichia coli isolates in diarrheal disease, we performed a study with 513 Venezuelan infants with diarrhea and 241 age-matched controls to determine the prevalence of enteropathogenic E. coli (enteroadherent E. coli, enterotoxigenic E. coli, enteroinvasive E. coli, and enterohemorrhagic E. coli) and their correlation with O:H serotypes. E. coli isolates exhibiting localized and aggregative adherence in the HEp-2 cell assay were significantly more frequently isolated from the patients (8.5 and 26.9%, respectively) than from the controls (1.7 and 15%, respectively). This difference was significant for the group 0 to 2 months of age but for older infants. Regardless of age, E. coli isolates with diffuse adherence were found at similar frequencies in both the patients and the controls. A striking correlation between classic O serogroups and localized adherence was also observed. These findings confirm the pathogenic role of E. coli with localized and aggregative adherence in diarrheal disease, as well as the epidemiological importance of O:H serotyping for characterizing localized-adhering E. coli.

Information concerning the disease burden of rotavirus, particularly in developing countries, has important implications for the use and for monitoring the impact of rotavirus vaccines. Although rotavirus has been recognized as the most frequent cause of hospitalization in the world, national estimates and specific information about the incidence of hospitalization for rotavirus gastroenteritis are very limited. Consequently, estimates of the incidence of hospitalization among children during the first 2 years of life in Venezuela were determined by extrapolation of data from a community-based study carried out in Caracas.

Some aspects of rotavirus humoral immunity were assessed on the basis of distinguishing serotype-specific specificities (VP4/VP7) by using rotavirus reassortants, human and animal strains in neutralization assays in serum samples obtained during the acute phase, and 1, 6 and 12 months after primary natural infection. In this study, all the infecting virus strains were characterized as G type and some also as P type. Primary natural infection induces a significantly greater homotypic neutralization response than heterotypic response. In addition, there was no significant difference in the number of homotypic or heterotypic responses following reinfection. Transplacentally acquired homotypic antibodies were associated with protection against dehydration during rotavirus gastroenteritis.

The temporal distribution and clinical severity of rotavirus VP7 serotypes 1, 2, 3, and 4 recovered from 427 Venezuelan children with acute gastroenteritis over a period of 11 years were studied. Rotavirus VP7 serotype was established by ELISA serotyping in 298 (69.78%) of the specimens while the serotype of the remaining 129 (30.21%) samples could not be determined. Of the specimens typed, 85 (19.90% of the total) were serotype 1, 43 (10.07%) were serotype 2, 105 (24.59%) were serotype 3, and 65 (15.22%) were serotype 4. Yearly changes in the frequency of individual serotypes were observed. The predominance of a single serotype with minor contribution from others was noted every year. In this study, serotype 1 appears to induce a less severe illness in comparison with serotypes 2, 3, and 4. No apparent association between the proportion of each serotype and the children's age were found.

90 Venezuelan infants aged 10-20 weeks were randomly allocated to four groups which received one of the following: the M37 vaccine (1 x 10 4 pfu [plaque-forming units]); quadrivalent rotavirus vaccine (1 × 10 4 pfu each of serotype 3 rhesus rotavirus [RRV] and human rotavirus-RRV reassortants of serotypes 1, 2, and 4); balanced quadrivalent vaccine consisting of 1 × 10 4 pfu of serotype 1 and 3 components but 5 × 10 4 pfu of serotype 2 and 4 components; or placebo. The frequencies of transient febrile responses in these four groups were 20%, 27%, 30%, and 9%. 50% of 22 infants tested who received M37 vaccine showed a serum rotavirus IgA antibody response, compared with 74% of the 23 quadrivalent and 86% of the 22 balanced-quadrivalent recipients. 64% of the M37 recipients showed a neutralising antibody response to M37; 27% showed such responses to human serotype 1 Wa strain and 27% to serotype 4 neonatal strain ST3. 17-39% of the quadrivalent recipients and 27-41% of the balanced-quadrivalent recipients showed neutralising antibody responses to serotypes 1-4. 70-73% of the quadrivalent and balanced quadrivalent groups also showed neutralising antibody responses to RRV.

The genetic relatedness of 81 clinical rotavirus isolates to the human rotavirus prototype strains Wa (subgroup 2, serotype 1) and DS-1 (subgroup 1, serotype 2) was examined by RNA hybridization techniques. Labeled single-stranded (+) transcripts of Wa or DS-1 virus were incubated with denatured genomic rotaviral RNAs, and the resulting hybrids were subjected to gel electrophoresis and autoradiography. Nineteen of the specimens contained subgroup 1 rotavirus with a "short" RNA migration pattern. These viruses were found to be closely related to the DS-1 strain and were associated with illness of short duration. The remaining 62 isolates belonged to subgroup 2 and exhibited a "long" RNA migration pattern. Fifty-four of these isolates exhibited significant hybridization with the Wa strain probe. Four isolates yielded multiple hybrid bands with the Wa probe but also possessed at least one gene segment homologous to the DS-1 strain. The remaining four subgroup 2 rotaviruses did not exhibit significant homology in the form of labeled hybrid bands when tested with either the Wa or DS-1 probe. These findings suggest that most clinical rotavirus isolates belong to one of two human rotavirus "families" defined as Wa-like or DS-1-like. Our observations also suggest that reassortment occurs in vivo between rotaviruses belonging to the two human rotavirus "families" and that there are one or more additional families of human rotavirus.

We have developed a hybridization assay that permits distinction of rotavirus serotypes 1, 2, 3, and 4. The serotype of rotaviruses from stool samples or tissue culture was recognized by hybridization of specific probes to (i) blots of viral double-stranded RNAs electrophoresed in agarose gels (Northern blots) or (ii) heat-denatured double-stranded RNAs directly dotted on nylon membranes. The probes consisted of 32P-labeled cDNA synthesized by reverse transcription of in vitro derived rotavirus mRNA from rotavirus serotypes 1 to 4. To prepare these probes, mRNAs were primed with a 17-mer nucleotide common to all four serotypes whose sequence is complementary to bases 375 to 391 of the rotavirus gene encoding the VP7 glycoprotein (gene 8 or 9 depending on the rotavirus strain). The resulting downstream transcripts encompassed areas of major sequence divergence among the four serotypes. Hybridization at high stringency (50 degrees C, 50% formamide, 4 x SSC [1 x SSC is 0.15 M NaCl plus 0.015 M sodium citrate]) was performed for 16 to 48 h. Autoradiograms of the washed membranes allowed recognition of the rotavirus serotype present in the blotted or dotted specimens since each of them hybridized preferentially to one of the four probes. Twenty-four laboratory specimens and 103 clinical specimens from Washington, D.C., Venezuela, and Chile were "serotyped" with this assay. The results were similar to those obtained with a monoclonal antibody serotyping assay.

The reactogenicity and antigenicity of the rhesus rotavirus vaccine, strain MMU18006, developed at the Laboratory of Infectious Diseases (National Institute of Allergy and Infectious Diseases, National Institutes of Health) were examined in a double blind, placebo-controlled study of 40 newborn infants in Caracas, Venezuela. The children were observed for the first few days after birth in the hospital nursery and by home visits for 10 days after vaccination to detect any adverse reactions. No reactions could be attributed to the vaccine. Serologic responses to the vaccine were evaluated in paired sera obtained at birth (cord blood) and 4 weeks after vaccination. Serologic responses to the vaccine were not observed by complement fixation, neutralization or a rhesus rotavirus VP7 epitope-specific competition assay. However, such responses were found in 9 of 14 tested infants by an immunoglobulin A-specific enzyme-linked immunosorbent assay. Seventeen of the 20 vaccinees also shed rhesus rotavirus vaccine in stool during the postvaccination period.

Using an intracellular cytokine assay, we recently showed that the frequencies of rotavirus (RV)-specific CD4(+) and CD8(+) T cells secreting INFgamma, circulating in RV infected and healthy adults, are very low compared to the frequencies of circulating cytomegalovirus (CMV) reactive T cells in comparable individuals. In children with acute RV infection, these T cells were barely or not detectable. In the present study, an ELISPOT assay enabled detection of circulating RV-specific INFgamma-secreting cells in children with RV diarrhea but not in children with non-RV diarrhea without evidence of a previous RV infection. Using microbead-enriched CD4(+) and CD8(+) T cell subsets, IFNgamma-secreting RV-specific CD8(+) but not CD4(+) T cells were detected in recently infected children. Using the same approach, both CD4(+) and CD8(+) RV-specific T cells were detected in healthy adults. Furthermore, stimulation of purified subsets of PBMC that express lymphocyte homing receptors demonstrated that RV-specific INFgamma-secreting CD4(+) T cells from adult volunteers preferentially express the intestinal homing receptor alpha4beta7, but not the peripheral lymph node homing receptor L-selectin. In contrast, CMV-specific INFgamma-secreting CD4(+) T cells preferentially express L-selectin but not alpha4beta7. These results suggest that the expression of homing receptors on virus-specific T cells depends on the organ where these cells were originally stimulated and that their capacity to secrete INFgamma is independent of the expression of these homing receptors.