Biology and molecular characterization of necroviruses

Three necroviruses, family Tombusviridae, are known to infect Olea europaea L., Olive latent virus 1 (OLV-1), Olive mild mosaic virus (OMMV) and Tobacco necrosis virus D (TNV-D), often asymptomatically. OLV-1 has a wide geographical distribution whereas that of OMMVand TNV-D is not known as their discovery is recent, dating from 2005 and 2009, respectively. OMMV has received special attention because of its origin, likely to have resulted from a recombination event between TNV-D, with which it shares a coat protein (CP) amino acid identity of 86% and OLV-1 sharing an RNA-dependent RNA polymerase amino acid identity of 91%. Due to these properties OMMV and TNV-D are indistinguishable by serology or Reverse Transcription – Polymerase Chain Reaction assays that use primers hybridizing in the CP gene region, rendering ambiguous earlier TNV-D identifications based on such tests. OLV-1 and OMMV are transmitted through the soil to roots of host plants in the absence of any vectors but the transmission rate of OMMV increases to more than double in the presence of Olpidium brassicae zoospores that vector this virus in an in vitro manner. A point mutation leading to an amino acid substitution located in an inner region of the virion is responsible for the loss of biologically assisted transmission, probably by altering the particle conformation rendering the binding site inaccessible to the zoospore receptors. The field occurrences of necrovirus multiple infections are common and increase the chances for recombination events, the epidemiological implications of which are still uncertain. The biological and molecular properties of the necroviruses identified in olive trees are the main focus of this review

Cellular eIF2B subunit localisation: implications for the integrated stress response and its control by small molecule drugs

eIF2 is a G protein critical for translation. It is tightly regulated in the integrated stress response (ISR) via phosphorylation of eIF2α and the subsequent control of eIF2B, a multisubunit guanine nucleotide exchange factor (GEF). Through studying the localisation of eIF2B subunits we identified cytoplasmic eIF2B bodies in mammalian cells. We highlight a relationship between body size and the eIF2B subunits localising to them; larger bodies contain all subunits and smaller bodies contain predominantly catalytic subunits. eIF2 localises to eIF2B bodies and shuttles within these bodies in a manner which correlates with eIF2B activity. Upon stress eIF2α-P localises predominately to larger bodies and results in a decreased shuttling of eIF2. Interestingly drugs which inhibit the ISR can rescue eIF2 shuttling in a manner correlating to levels of eIF2α-P. In contrast, smaller bodies show increased eIF2 shuttling in response to stress, which is accompanied by the localisation of eIF2Bδ to these bodies, suggesting the formation of a novel trimeric complex of eIF2B. This response is mimicked by ISR inhibiting drugs, providing insight into their potential mechanism of action. This study provides evidence that the composition and function of mammalian eIF2B bodies is regulated by the ISR and drugs which control it

D etection of sweet potato virus C, sweet potato virus 2 and sweet potato feathery mottle virus in Portugal

Field sweet potato plants showing virus-like symptoms, as stunting, leaf distortion, mosaic and chlorosis, were collected in southwest Portugal and tested for the presence of four potyviruses, sweet potato virus C (SPVC), sweet potato virus 2 (SPV2), sweet potato feathery mottle virus (SPFMV), sweet potato virus G (SPVG), and the crinivirus sweet potato chlorotic stunt virus (SPCSV). DsRNA fractions were extracted from symptomatic leaves and used as templates in single and multiplex RT-PCR assays using previously described specific primers for each analyzed virus. The amplified reaction products for SPVC, SPV2 and SPFMV were of expected size, and direct sequencing of PCR products revealed that they correspond to the coat protein gene (CP) and showed 98%, 99% and 99% identity, respectively, to those viruses. Comparison of the CP genomic and amino acid sequences of the Portuguese viral isolates recovered here with those of ten other sequences of isolates obtained in different countries retrieved from the GenBank showed very few differences. The application of the RT-PCR assays revealed for the first time the presence of SPVC and SPFMV in the sweet potato crop in Portugal, the absence of SPVG and SPCSV in tested plants, as well as the occurrence of triple virus infections under field conditions

Complete nucleotide sequence of an Olive latent virus 1 isolate from olive trees

Olive latent virus 1 (OLV-1) is a necrovirus belonging to the familyTombusviridae. It is a small icosahedral plant virus, which encapsidates a single stranded positivesense RNA. This virus was first isolated from symptomless olive trees in Italy [7] and afterwards in Jordan and Portugal [10, 4]. OLV-1 was also isolated from symptomatic hosts, such as citrus trees in Turkey [11] and tulips in Japan [9]. Up to now, only one complete genome sequence of an OLV-1 citrus isolate has been determined [8]. This report describes the first full genomic sequence of an OLV-1 isolated from olive trees

Risco de Recidiva a 5 Anos Após Excisão Convencional de um Carcinoma Basocelular

Introduction: Basal cell carcinomas are mostly treated surgically, mostly by surgery with postoperative histopathologic margin evaluation (“conventional surgery”), but large long-term data regarding recurrence by completeness of excisions is limited. Methods: Retrospective cohort study of basal cell carcinomas treated by conventional surgery at different medical specialties in a large tertiary centre, between 2008 and 2014. Survival analysis with a Cox proportional-hazards was performed, stratified by completeness of excision (complete excision/incomplete excision) and adjusted to several potentially confounding covariates. Results: A total of 2876 basal cell carcinomas were identified, of which 2306 (2100 primary, 206 recurrent) were considered eligible for analysis. During the 5-years of follow-up, there were 80 (4%) recurrences among 1980 complete excisions (16/1000 cases-year) and 83 (23.9%) recurrences among 348 incomplete excisions (100/1000 cases-year). Survival analysis was performed with multivariable adjustment. In the final adjusted model, we identified an association between relapse and re-intervention on recurrent tumors [adjusted Hazard Ratio (HR) 2.20 (95% Confidence interval (IC), 1.26-3.84), p=0.006], a wrong preoperative clinical diagnosis/surgery devoid of preoperative biopsy [adjusted HR 2.75 (95% CI, 1.68-4.5), p<0.001], treatment prior to 2012 [adjusted HR 1.47 (95% IC, 1.06-2.05), p<0.021] and surgery on a high-risk location, accordingly to the NCCN stratification [adjusted HR 2.18 (95% CI, 1.08-4.40), p<0.030]. By specific anatomic location, the likelihood of recurrence was especially high in the nose [adjusted HR 3.18 (95% CI 1.71-5.87), p<0.001] and eyelids [adjusted HR 3.08 (95% CI, 1.32-7.17), p=0.009]. There was also a trend towards higher recurrence in aggressive histological subtypes [adjusted HR 1.43 (95% CI 0.99-2.07), p<0.058]. Conclusion: Recurrent basal cell carcinomas, regardless of location, and primary basal cell carcinomas on high-risk locations of the face, especially on the eyelids and nose, should be considered to have a higher and independent likelihood of recurrence, even on “complete excisions” evaluated by histopathology. On the other hand, wait-andsee approaches in incompletely excised BCCs should be considered against a significant 5-year risk of relapse (1 in 10 lesions).Introdução: O tratamento dos carcinomas basocelulares é maioritariamente cirúrgico, sobretudo por cirurgia com avaliação histopatológica pós-operatória da margem (cirurgia convencional), mas os dados a longo-prazo relativos a recidiva de acordo com o resultado histológico da margem (excisão completa versus excisão incompleta, mantida em follow-up) são limitados. Métodos: Estudo coorte retrospetivo dos carcinomas basocelulares tratados por cirurgia convencional e por diferentes especialidades médico-cirúrgicas num centro terciário, entre 2008 e 2014. Realizou-se uma análise multivariada com uma regressão de Cox, estratificada pelo resultado da avaliação histológica da margem (excisão completa/incompleta) e ajustada a várias variáveis recolhidas. Resultados: Um total de 2876 carcinomas basocelulares foram identificados, dos quais 2306 [2100 primários, 206 recidivantes (primeira recidiva)] foram considerados elegíveis para análise. Nos 5 anos de follow-up, verificaram-se 80 (4%) recidivas entre os 1959 tumores completamente excisados (16/1000 casos-ano), contrastando com 83 (23,9%) recidivas em 347 excisões incompletas (100/1000 casos-ano). Foi realizada uma análise de sobrevida ajustada. No modelo final, ajustado, multivariado, foi identificada associação entre recidiva e intervenção cirúrgica a tumores recorrentes [hazard ratio (HR) ajustado 2,20 (Intervalo confiança (IC) 95%, 1,26-3,84), p=0,006], cirurgia com diagnóstico pré-operatório errado/ausência de realização de biópsia prévia [HR ajustado 2,75 (IC 95%, 1,68-4,5), p<0,001], tratamento prévio a 2012 [HR ajustado 1,47 (CI 95%, 1,06- 2,05), p<0,021] e cirurgia em localização de alto-risco, de acordo com a classificação NCCN [HR ajustado 2,18 (IC 95%, 1,08- 4,40), p<0,030]. Por localização anatómica específica, a probabilidade de recidiva a longo-prazo é especialmente elevada se a cirurgia for na pirâmide nasal [HR ajustado 3,18 (IC 95%, 1,71-5,87), p<0,001] ou nas pálpebras [HR ajustado 3,08 (CI 95%, 1,32-7,17), p=0,009]. Verificou-se também uma tendência para maior recidiva nos subtipos histológicos agressivos [HR ajustado 1,43 (IC 95%, 0,99-2,07), p<0,058]. Conclusão: Os carcinomas basocelulares recorrentes, independentemente da localização, e os carcinomas basocelulares primários em localizações de alto-risco da face, particularmente na pirâmide nasal e nas pálpebras, determinam um risco de recidiva superior e independente a longo-prazo, mesmo nas excisões “completas”. Por outro lado, as estratégias wait-and-see nos carcinomas basocelulares incompletamente excisados devem ponderar o risco de recidiva aos 5 anos (1 in 10 lesões)

Dor pós-operatória em mulheres submetidas à cesariana

Objective: To estimate the incidence, intensity and quality for pain post-cesarean section. Methods: Cross-sectional study. A total of 1062 post-cesarean section women were interviewed in immediates pre- and post-operative. Pain intensity and quality were evaluated through the Numerical Rating Scale (0-10) and McGill Pain Questionnaire-SF. The variables were explored by descriptive measures and the incidence of postoperative pain calculated with a 95% confidence interval. Results: The incidence of pain was 92,7% (IC 95%: 90,9 - 94,2). The average level of pain intensity at the time of worst pain was 6,6 (dp=2,2). The descriptors more frequently chosen were “aching” (91.6%) “tender” (70.0%) and “throbbing” (56.1%). Conclusions: High-intensity postoperative pain is a reality for post-cesarean section women, showing the importance of pain assessment for implementation of curative and preventive actions to reduce losses in the recovery of women.Objetivo: Estimar la incidencia, intensidad y calidad del dolor postoperatorio en mujeres sometidas a cesárea. Método: Estudio de corte transversal. Fueron entrevistadas 1062 mujeres sometidas a cesárea en el periodo pre y post cirurgía inmediatos. La intensidad del dolor y la calidad fueron evaluadas por la Escala Numérica del Dolor (0-10) y el Test de Dolor de McGill. Las variables se analizaron utilizando medidas descriptivas y la incidencia de dolor postoperatorio calculado con un Intervalo de Confianza del 95%. Resultados: La incidencia de dolor fue del 92,7% (IC 95%: 90,9 - 94,2) y las puntuaciones de intensidad media en el momento de peor dolor, igual a 6,6 (dp=2,2) . Los descriptores más frecuentes escogidos fueron "dolorida" (91,6%), "dolorida a la palpación" (70,0%) y "palpitante" (56,1%). Conclusiones: El dolor postoperatorio de alta intensidad es una realidad en este grupo, señalando la importancia de la evaluación del dolor para la ejecución de las acciones curativas y preventivas para reducir las pérdidas en la recuperación de las mujeresObjetivo: Estimar a incidência, intensidade e qualidade da dor pós-operatória em mulheres submetidas à cesariana. Método: Estudo tipo corte transversal. Foram entrevistadas 1062 mulheres submetidas à cesariana nos períodos pré e pós-operatório imediatos. A intensidade e qualidade da dor foram avaliadas por meio da Escala Numérica de Dor (0-10) e Questionário de Dor de McGill. As variáveis foram exploradas por meio de medidas descritivas e a incidência de dor pós-operatória calculada com Intervalo de Confiança de 95%. Resultados: A incidência de dor foi de 92,7% (IC 95%: 90,9 - 94,2). A média dos escores de intensidade, no momento de pior dor, igual a 6,6 (dp=2,2). Os descritores escolhidos com maior frequência foram “dolorida” (91,6%), “dolorida à palpação” (70,0%) e “latejante” (56,1%). Conclusões: A dor pós-operatória de elevada intensidade é uma realidade neste grupo, apontando a importância da avaliação da dor para implementação de ações curativas e preventivas que reduzam prejuízos na recuperação das mulheres

Effect of the inherent variation in the mineral concentration of alfafa cultivars on aphid populations.

Plants have inherent variability of mineral content which affects their physiology and consequently the herbivorous insects feeding on them. Besides, insects need considerable amounts of potassium, phosphorus and magnesium in their diets, whereas little calcium, sodium and chloride are required. In this study, the inherent variation on mineral (Ca, S, Mg, N, P, K and also C:N ratio) concentrations and aphid (Acyrthosiphon spp., Therioaphis maculata, Aphis craccivora) populations on three alfalfa (Medicago sativa) cultivars (P3; Crioula, the most widely cultivated in Brazil, and CUF 101, an aphid-resistant) were studied between September/1997 and August/1998. A significant variation on mineral concentrations and aphid populations was observed among different sampling times and cultivars. The correlations between C:N ratio, Mg, N, P and S concentrations and aphid density variation suggest that the mineral status affects aphid population dynamics under field conditions

Multi-year statistical and modeling analysis of submicrometer aerosol number size distributions at a rain forest site in Amazonia

The Amazon Basin is a unique region to study atmospheric aerosols, given their relevance for the regional hydrological cycle and the large uncertainty of their sources. Multi-year datasets are crucial when contrasting periods of natural conditions and periods influenced by anthropogenic emissions. In the wet season, biogenic sources and processes prevail, and the Amazonian atmospheric composition resembles preindustrial conditions. In the dry season, the basin is influenced by widespread biomass burning emissions. This work reports multi-year observations of high time resolution submicrometer (10-600 nm) particle number size distributions at a rain forest site in Amazonia (TT34 tower, 60 km NW from Manaus city), between 2008 and 2010 and 2012 and 2014. The median particle number concentration was 403 cm(-3) in the wet season and 1254 cm(-3) in the dry season. The Aitken mode (similar to 30-100 nm in diameter) was prominent during the wet season, while the accumulation mode (similar to 100-600 nm in diameter) dominated the particle size spectra during the dry season. Cluster analysis identified groups of aerosol number size distributions influenced by convective downdrafts, nucleation events and fresh biomass burning emissions. New particle formation and subsequent growth was rarely observed during the 749 days of observations, similar to previous observations in the Amazon Basin. A stationary 1-D column model (ADCHEM Aerosol Dynamics, gas and particle phase CHEMistry and radiative transfer model) was used to assess the importance of the processes behind the observed diurnal particle size distribution trends. Three major particle source types are required in the model to reproduce the observations: (i) a surface source of particles in the evening, possibly related to primary biological emissions; (ii) entrainment of accumulation mode aerosols in the morning; and (iii) convective downdrafts transporting Aitken mode particles into the boundary layer mostly during the afternoon. The latter process has the largest influence on the modeled particle number size distributions. However, convective downdrafts are often associated with rain and, thus, act as both a source of Aitken mode particles and a sink of accumulation mode particles, causing a net reduction in the median total particle number concentrations in the surface layer. Our study shows that the combination of the three mentioned particle sources is essential to sustain particle number concentrations in Amazonia.Peer reviewe

Manipulating the number of players and targets in team sports : small-sided games during physical education classes

This study aimed to examine the effects of players and targets number variation on technical, tactical and physiological indicators in basketball, handball and indoor football. Nine 8th grade students (age 13.3 ± 0.7 years, stature 166.8 ± 3.8 cm, body mass 55.3 ± 4.2 kg) participated in four small-sided games (SSG) for each sport. Despite the differences between the sports, similar average values for technical and tactical indicators were found in both basketball and indoor football SSG. An increase in the number of players affected the duration of unsuccessful offensive phases, the number of passes per offensive phase, successful passes per offensive phase and decreased ball possession per player. Whilst all SSG in this study imposed high physiological demands, 3-a-side games played with one target elicited the greatest responses. In conclusion, the 3-a-side SSG promoted similar technical-tactical indicators and higher physiological demands than 4-a-side SSG.Este estudio tuvo como objetivo examinar los efectos del número de jugadores y metas en la variación de los indicadores técnicos, tácticos y fisiológicos en baloncesto, balonmano y fútbol sala. Nueve estudiantes del octavo grado (edad 13.3 ± 0.7 años, estatura 166.8 ± 3.8 cm, masa corporal 55.3 ± 4.2 kg) participarón en cuatro juegos reducidos (SSG) para cada deporte. A pesar de las diferencias entre los deportes, se encontrarón valores medios similares en los indicadores técnicos y tácticos tanto en los juegos reducidos del baloncesto como en los de fútbol sala. El aumento de los jugadores ha provocado el aumento de la duración de la fase ofensiva sin éxito, número de pases por fase ofensiva, pases acertados por fase ofensiva y ha provocado una disminución de la posesión del balón por jugador. Mientras la totalidad de juegos reducidos utilizados en este estudio han impuesto un elevada demanda fisiológica, los juegos reducidos de 3x3 jugados con una meta provocaron las mayores respuestas. En conclusión, los juegos reducidos de 3x3 ha inducido valores similares en los indicadores técnico-tácticos, pero demandas fisiológicas más altas que los juegos reducidos de 4x4.Este estudo teve como objectivo analisar os efeitos do número de jogadores e dos objectivos na variação dos indicadores técnicos, tácticos e fisiológicos no basquetebol, andebol e futsal. Nove estudantes do oitavo ano (idade 13.3 ± 0.7 anos, estatura 166.8 ± 3.8 cm, massa corporal 55.3 ± 4.2 kg) participaram em quatro jogos reduzidos (SSG) para cada modalidade. Apesar das diferenças entre os desportos, foram verificados valores médios similares nos indicadores técnicos e tácticos nos jogos reduzidos de basquetebol e futsal. O aumento dos jogadores provocou o aumento da duração da fase ofensiva sem êxito, número de passes por fase ofensiva, passes certos por fase ofensiva e provocou uma diminuição da posse de bola por jogador. Enquanto a totalidade de jogos reduzidos utilizados neste estudo implicou uma elevada exigência fisiológica, os jogos reduzidos de 3X3 jogados com um objectivo provocaram as maiores respostas. Em suma, os jogos reduzidos de 3x3 induziram valores similares nos indicadores técnico-tácticos, mas exigências fisiológicas mais elevadas que os jogos reduzidos de 4X4