The movement proteins (NSm) of distinct tospoviruses peripherally associate with cellular membranes and interact with homologous and heterologous NSm and nucleocapsid proteins.

Tospovirus is the only genus containing virus species which infect plants in the Bunyaviridae family. The aims of this study were to understand the in vivo membrane association of the movement protein (NSm) of the tospovirus species Bean necrotic mosaic virus, Chrysanthemum stem necrosis virus, Tomato chlorotic spot virus and Tomato spotted wilt virus and the homologous and heterologous interactions among NSm and nucleocapsid protein (N). The results obtained by bimolecular fluorescence complementation (BiFC) assay and chemical treatments after membrane fractionation revealed that the four NSm proteins are associated with the biological membranes with the N- and C-termini oriented to the cytoplasm. BiFC analysis for protein-protein interactions showed: i) dimer formation for all NSm and N proteins; ii) interaction between NSm and the cognate N and iii) heterologous interactions between the NSm and N proteins. The implications of these interactions in the life cycle of tospoviruses are discussed.

Ilarviruses of Prunus spp.: a continued concern for fruit trees.

Prunus spp. are affected by a large number of viruses, causing significant economic losses through either direct or indirect damage, which results in reduced yield and fruit quality. Among these viruses, members of the genus Ilarvirus (isometric labile ringspot viruses) occupy a significant position due to their distribution worldwide. Although symptoms caused by these types of viruses were reported early in the last century, their molecular characterization was not achieved until the 1990s, much later than for other agronomically relevant viruses. This was mainly due to the characteristic liability of virus particles in tissue extracts. In addition, ilarviruses, together with Alfalfa mosaic virus, are unique among plant viruses in that they require a few molecules of the coat protein in the inoculum in order to be infectious, a phenomenon known as genome activation. Another factor that has made the study of this group of viruses difficult is that infectious clones have been obtained only for the type member of the genus, Tobacco streak virus. Four ilarviruses, Prunus necrotic ringspot virus, Prune dwarf virus, Apple mosaic virus, and American plum line pattern virus, are pathogens of the main cultivated fruit trees. As stated in the 9th Report of the International Committee on Taxonomy of Viruses, virions of this genus are "unpromising subjects for the raising of good antisera." With the advent of molecular approaches for their detection and characterization, it has been possible to get a more precise view of their prevalence and genome organization. This review updates our knowledge on the incidence, genome organization and expression, genetic diversity, modes of transmission, and diagnosis, as well as control of this peculiar group of viruses affecting fruit trees.

First Report of Apricot latent virus and Plum bark necrosis stem pitting-associated virus in Apricot from Spain.

Representing 2% of world production, 20,000 ha of apricot (Prunus armeniaca L.), are cultivated in Spain, primarily in the southeast. A survey was conducted during the spring of 2008 in orchards in the region of Murcia to assess the incidence of several stone fruit viruses. Leaf and fruit samples from 160 trees from 40 orchards were collected randomly for reverse transcription (RT)-PCR analysis. Total RNA extracted (3) from leaves and fruits was tested by a multiplex one-step RT-PCR protocol with a mix of primers that detect eight distinct viruses (4). Amplicons of 250 bp expected for Plum bark necrosis stem pitting-associated virus (PBNSPaV), corresponding to part of the heat shock 70 protein gene, were obtained from four trees and amplicons of 700 bp expected for Apricot latent virus (ApLV), corresponding to part of the coat protein (CP) gene, were obtained from two trees. In all cases, amplicons were obtained using RNA extracted from leaf and fruit tissues. RT-PCR results were confirmed by uniplex RT-PCR with primers specific for each virus and dot-blot hybridization with virus-specific digoxygenin-labeled RNA probes (2). To further characterize the new viruses, we designed primers to amplify specifically the CP gene of ApLV (5'-CCCGACCATGGCTACAAGC-3' and 5'-TTGCCGTCCCGATTAGGTTG-3') and the minor CP gene of PBNSPaV (5'-GAACAAACTACAGCAGCACC-3' and 5'-CAAGGGTAGGACGGGTAACGC-3'). Amplicons of 1,500 and 950 bp corresponding to the ApLV and PBNSPaV CP genes, respectively, were purified from agarose gels and cloned in the pTZ57R plasmid (Fermentas, Burlington, Ontario, Canada). Blastp analysis of the full-length ApLV CP sequence from one infected tree (GenBank Accession No. GQ919051) revealed 86% amino acid (aa) similarity to the single full-length ApLV CP sequence available (No. AAC16234) and 79 and 66.9% similarity to Peach sooty ringspot virus (No. AAG48314) and Apple stem pitting virus (No. NP604468), respectively. Identity/similarity analysis of the full-length PBNSPaV minor CP genes using the Matrix Global Alignment Tool software, version 2.02 (1), revealed 98.8 to 99.6% aa similarity between the Spanish PBNSPaV isolates (Nos. GQ919047, GQ919048, GQ919049, and GQ919050) and 97.1 to 97.4% with the PBNSPaV isolate from the United States (No. EF546442). None of the six infected trees were associated with any particular field symptoms. Five infected trees were cv. Búlida and one was native cv. Murciana, which was infected with ApLV. All infected trees were located in geographically separated orchards. The incidence of ApLV and PBNSPaV was 1.25 and 2.5%, respectively. The low incidence of both viruses together with the scattered geographic distribution could be due to the recent introduction of virus-contaminated plants, although we cannot exclude that it is a consequence of a low dissemination rate. Even though no symptoms were observed, we cannot discard that the infection could affect fruit production or flowering or even cause a synergistic effect in mixed infection with other stone fruit viruses, a risk especially relevant considering the total area of cultivated apricot. To our knowledge, this is the first report of ApLV and PBNSPaV in Spain. References: (1) J. J. Campanella et al. BMC Bioinformatics 4:29, 2003. (2) M. C. Herranz et al. J. Virol. Methods 124:49, 2005. (3) D. J. Mackenzie et al. Plant Dis. 81:222, 1997. (4) J. A. Sánchez-Navarro et al. Eur. J. Plant Pathol. 111:77, 2005.

Multiplex polymerase chain reaction (PCR) and real-time multiplex PCR for the simultaneous detection of plant viruses.

Multiplex Polymerase Chain Reaction (PCR) can be used for the simultaneous detection of plant viruses. Multiple primer pairs or polyvalent primer pairs can be used to detect and identify several viruses in a single PCR.

First Report of Avocado sunblotch viroid in Avocado from Michoacán, México.

The State of Michoacán, México cultivates approximately 100,000 ha of avocados (Persea americana M.) (4). During a survey from 2006 to 2007 in cv. Hass avocado groves in Tingambato County, in the State of Michoacán, deep yellow spots and streaks, which sometimes became necrotic or reddish, were observed on the skin of fruits and the pulp of the fruit also showed big yellow spots. Some young shoots developed fine, yellow streaks, and leaves of symptomatic trees sometimes showed irregular, white-to-yellow spots. These symptoms were similar to those recorded for Avocado sunblotch viroid (ASBVd) (3). To determine if ABSVd was associated with these symptoms, total RNA extracted (1) from the skin and pulp of symptomatic and asymptomatic fruits and also from leaves and bark of shoots from five trees collected in a commercial plot in Tingambato County was tested by a one-step reverse transcription (RT)-PCR protocol using one primer pair to amplify specifically the complete ASBVd genome sequence (3). All 30 samples of skin and pulp of fruits, leaves, and cortex of shoots from symptomatic trees yielded two PCR fragments with estimated sizes of 250 and 500 base pairs (bp) corresponding to the putative monomeric and dimeric forms of ASBVd, respectively. The 500-bp RT-PCR fragments obtained from the different samples were purified from an agarose gel and cloned. The 249-bp nucleotide sequence of the ASBVd genomic monomer was determined using the clones from the fruit skin from sample Arb No. 3 (GenBank Accession No. EU888588), pulp from sample Arb No. 5 (GenBank Accession No. EU888590), leaves from samples Arb No. 15 (GenBank Accession No. EU888589) and Arb No. 8 (GenBank Accession Nos. EU888591 and EU888592), and cortex of shoots from sample Arb No. 16 (GenBank Accession Nos. EU888593, EU888594, EU888595, EU888596, and EU888597). BLAST analysis of the ASBVd sequences showed a range of 98 to 100% nucleotide identity to ASBVd sequences (GenBank Accession Nos. AF404064, AF404051, or AF229821). A clone of the Michoacán ASBVd (GenBank Accession No. EU888593) was used to synthesize a Dig-High Prime-UTP-T7 (Roche, Mannheim, Germany) fluorescent riboprobe complementary to the ASBVd plus strand to perform a dot-blot analysis as described previously (2). All ASBVd samples positive by RT-PCR gave a strong signal in the dot-blot analysis. This riboprobe will be used to index the ASBVd in other commercial avocado groves in Michoacán. To our knowledge, this is the first report of ASBVd in Michoacán, México. References: (1) D. J. Mackenzie et al. Plant Dis. 81:222, 1997. (2) J. A. Sánchez-Navarro et al. Plant Pathol. 47:780, 1998. (3) R. J. Schnell et al. Plant Dis. 81:1023, 1997. (4) D. Téliz and A. Mora. El aguacate y su Manejo Integrado. Mundiprensa, Mexico City, 2007.

Adenopatía axilar y fiebre / No disponible

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Fractura-luxación de hombro con fragmento intrapulmonar de la cabeza humeral / Fracture-dislocation of the shoulder with intrapulmonary location of the humeral head

Se presenta el caso de un varón de cincuenta años que sufrió una caída accidental de un caballo, recibiendo el impacto del animal sobre su cuerpo en la caída, produciéndose una fractura-luxación de la extremidad proximal del húmero, con localización intrapulmonar de la cabeza humeral. El paciente necesitó la colocación de un tubo de drenaje torácico e ingresó en la Unidad de Cuidados Intensivos (UCI) hasta su estabilización, tras la cual fue intervenido quirúrgicamente mediante una artroplastia parcial de hombro y seguidamente se procedió a la extracción, a través de una toracotomía, del fragmento humeral de localización intraparenquimatosa

We present the case of a 50-year-old male who was injured as a result of a fortuitous fall from a horse. On falling, the patient impacted on the horse aad sustained a fracture of the proximal third of he humerus, with the humeral head ending up in an intrapulmonary location. The patient had to be administered thoracic drainage and was admitted into the ICU until he was stabilized. Subsequently, he was subjected to a partial shoulder arthroplasty and to a thoracotomy in order to extract the humeral fragment from its intraparenchymal location

Low genetic variability in the coat and movement proteins of American plum line pattern virus isolates from different geographic origins.

Until very recently isolates of American plum line pattern virus (APLPV) had not been reported from outside North America. The nucleotide sequences corresponding to the movement (MP) and coat (CP) proteins of eight APLPV isolates from five Mediterranean countries were determined. Sequence analysis showed that both MP and CP genes are highly conserved irrespective of geographic origin. The study of the distribution of synonymous and nonsynonymous changes along both open reading frames revealed that these proteins are under the effect of negative purifying selection. The MP and CP of APLPV possess most of the functional motifs described for other members of the genus Ilarvirus.

First Report of Tomato torrado virus on Weed Hosts in Spain.

Tomato torrado virus (ToTV) is a recently identified Picorna-like virus that causes "torrado disease" in tomatoes (4). Typical symptoms of "torrado disease" seen in tomato crops (Solanum lycopersicum L. formerly Lycopersicon esculentum L.) were initially defined as yellow areas at the base of the leaflet that later developed into necrotic spots that sometimes abscised, leaving holes in the leaflet. Other plants showed extensive necrosis progressing from the base to the tip of the leaflet. Fruits were distorted with necrotic lines on the surface that often cracked. Affected plants had a burnt-like appearance and the production was seriously reduced. These symptoms have been observed in tomato crops in Murcia (Spain) and the Canary Islands (Spain) (1). To identify possible alternative hosts that may serve as virus reservoirs, samples of 72 different common weed species were collected in greenhouses in Murcia and the Canary Islands where "torrado disease" symptoms were observed in tomatoes. Forty-seven showed virus-like symptoms and 25 were asymptomatic. Symptoms included mild mosaic, blistering, vein clearing, interveinal yellowing, yellow spots, necrosis, leaf distortion, and curling. Samples were analyzed by one-step reverse transcription (RT)-PCR using primers specific for ToTV to amplify 580 bp of the polyprotein region of RNA2 (3) and dot-blot hybridization with a digoxygenin-labeled RNA probe complementary to the same portion of the ToTV genome. Twenty-two of the 72 weed samples belonging to Amaranthus sp. (Amaranthaceae); Spergularia sp. (Caryophyllaceae); Atriplex sp., Chenopodium ambrosioides L., Chenopodium sp., and Halogetum sativus (Loef. ex L.) Moq. (Chenopodiaceae); Senebiera didyma Pers. (Cruciferae); Malva sp. (Malvacae); Polygonum sp. (Polygonaceae); and Nicotiana glauca Graham and Solanum nigrum L. (Solanaceae) were positive for ToTV by molecular hybridization (10 samples) and RT-PCR (22 samples, including the samples positive by molecular hybridization). PCR products obtained from Atriplex sp. (Canary Islands) and S. didyma (Murcia) were sequenced (GenBank Accessions EU090252 and EU090253). BLAST analysis showed 99% identity to ToTV RNA2 sequence (GenBank Accession DQ388880). Two tomato plants were positive for ToTV by RT-PCR after mechanical back-inoculation, although no symptoms were observed. This study showed ToTV infects common weeds present in Spanish tomato crops. Recently, Trialeurodes vaporariorum has been reported to transmit ToTV (2), although the efficiency of transmission is unknown. The vector-assisted transmission of ToTV could explain the infection of weeds in affected greenhouses. To our knowledge, this is the first report of natural infection of weeds by ToTV. References: (1) A. Alfaro-Fernández et al. Plant Dis. 91:1060, 2007. (2) H. Pospieszny et al. Plant Dis. 91:1364, 2007. (3) J. Van der Heuvel et al. Plant Virus Designated Tomato Torrado Virus. Online publication. World Intellectual Property Organization WO/2006/085749, 2006. (4) M. Verbeek et al. Arch. Virol. 152:881, 2007.

First Report of Tomato torrado virus in Tomato in the Canary Islands, Spain.

In 2003, greenhouse-grown tomato crops (Lycopersicon esculentum Mill.) in the Canary Islands (Spain) were observed showing an initial yellowing in defined areas at the base of the leaflet that later developed into necrotic spots or an extensive necrotic area progressing from the base to tip. Fruits were also affected, showing necrotic areas and often developing cracking. Generally, the plants that were affected seemed to be burnt, their growth was reduced, and the production level was seriously damaged. Similar symptoms have been observed in Murcia (Spain) since 2001, which have been recently associated with Tomato torrado virus (ToTV) infection (2). Twenty-two tomato samples showing "torrado disease" symptoms were collected from different greenhouses between 2003 and 2006 in Las Palmas (Canary Islands, Spain). To verify the identity of the disease, double-antibody sandwich (DAS)-ELISA was performed on leaf and fruit extracts of symptomatic plants using polyclonal antibodies specific to Potato virus Y (PVY), Tomato mosaic virus (ToMV), Tomato spotted wilt virus (TSWV) (Loewe Biochemica, Sauerlach, Germany), and Pepino mosaic virus (PepMV) (DSMZ, Braunschweig, Germany). Total RNA was extracted from the 22 tomato samples with the RNAwiz Extraction kit (Ambion, Huntingdon, United Kingdom) and tested using one-step reverse-transcription (RT)-PCR with the SuperScript Platinum Taq kit (Invitrogen Life Technologies, Barcelona, Spain) with primers specific to PepMV (1) and ToTV (2). All analyses included healthy tomato plants as negative controls. Five of the twenty-two tomato samples were positive for PepMV and negative for the other viruses tested by serological analysis. However, all 22 samples were positive in RT-PCR performed with the primers specific to ToTV segment RNA2. The RT-PCR assay to detect ToTV produced an amplicon of the expected size (580 bp). No amplification product was observed when healthy plants or a water control were used as a template in the RT-PCR reaction. The ToTV RT-PCR product was purified (High Pure PCR Product Purification kit, Roche Diagnostics, Mannheim, Germany) and sequenced. BLAST analysis of one sequence (GenBank Accession No. EF436286) showed 99% identity to ToTV RNA2 sequence (GenBank Accession No. DQ388880). To our knowledge, this is the first report of ToTV in the Canary Islands. References: (1) I. Pagán et al. Phytopathology 96:274, 2006. (2) M. Verbeek et al. Online Publication. doi:10.1007/s00705-006-0917-6. Arch. Virol., 2007.

[Brain mechanisms involved in decision-making]. / Mecanismos cerebrales de la toma de decisiones.

AIM: To review the studies on brain mechanisms in decision making within the framework of the somatic marker hypothesis, and based on experiments employing the Iowa Gambling Task. DEVELOPMENT: An overview of the somatic marker hypothesis is presented together with the review of the main results obtained from research in brain damaged patients, and normal subjects with functional neuroimaging studies, that have led to the identification of the neural structures involved in decision making in humans. CONCLUSIONS: The main region involved in decision making is the ventromedial prefrontal cortex, that integrates sensory, mnesic and emotional information relevant to the task. Other structures intervening in the various relevant processes in decision making are the amygdala (processing and encoding of the emotional signal and its association with contextual stimuli) and the cingulate cortex (process monitoring and response inhibition, especially in situations of uncertainty). The prefrontal dorsolateral cortex would also be involved through the necessary activation of the working memory in the decision making process, especially in the case of complex tasks.

Mecanismos cerebrales de la toma de decisiones / Brain mechanisms involved in decision-making

Objetivo. Revisar los estudios sobre los mecanismos cerebrales de la toma de decisiones en el marco de la hipótesis del marcador somático y basados experimentalmente en el empleo dela tarea de apuestas de Iowa (Iowa Gambling Task). Desarrollo. Se presenta la teoría del marcador somático y las características de la citada tarea de toma de decisiones y otras relacionadas. A continuación, se revisan los principales estudios llevados a cabo en personas con lesión cerebral y los procedentes de sujetos normales, con el empleo de neuroimagen funcional, que han hecho posible la identificación de las estructuras neurales implicadas en la toma de decisiones en humanos. Conclusiones. La principal región implicada es la corteza prefrontal ventromedial, donde se produce la integración de la información sensorial, amnésica y emocional necesaria para la tarea. Otras estructuras que intervienen en diferentes procesos relevantes para la toma de decisiones serían la amígdala (procesamiento y codificación de la señal emocional y su asociación con estímulos contextuales) y la corteza cingulada (monitorización del proceso e inhibición de respuesta, especialmente en situaciones de incertidumbre). La corteza prefrontal dorso lateral también participaría en este proceso debido a la necesaria activación de la memoria de trabajo en la toma de decisiones, en especial cuando la tarea es compleja (AU)

Aim. To review the studies on brain mechanisms in decision making within the framework of the somatic marker hypothesis, and based on experiments employing the Iowa Gambling Task. Development. An overview of the somatic marker hypothesis is presented together with the review of the main results obtained from research in brain damaged patients, and normal subjects with functional neuroimaging studies, that have led to the identification of the neural structures involved indecision making in humans. Conclusions. The main region involved in decision-making is the ventromedial prefrontal cortex, that integrates sensory, mnesic and emotional information relevant to the task. Other structures intervening in the various relevant processes in decision-making are the amygdala (processing and encoding of the emotional signal and its association with contextual stimuli) and the cingulate cortex (process monitoring and response inhibition, especially in situations of uncertainty). The prefrontal dorsolateral cortex would also be involved through the necessary activation of the working memory in the decision making process, especially in the case of complex tasks (AU)

Emotional response in patients with frontal brain damage: effects of affective valence and information content.

The authors investigated the role of the frontal lobes in the emotional response in 19 patients with brain damage and 23 control subjects. They studied the modulation of the startle blink reflex by affective pictures, and other autonomic responses. Patients showed a dissociation between the startle reflex and the affective valence ratings of the pictures, as a result of a low inhibition of the startle reflex by pleasant pictures. Pictures elicited lower skin conductance responses (SCRs) in patients than in controls, whereas the groups did not differ in the SCRs prompted by less significant acoustic stimuli. The findings point to the frontal lobe as a structure involved in the emotional response and in the physiological emotional arousal related to the complexity of the stimuli.

Cirugía torácica mínimamente invasiva: Presentación / No disponible

No disponible

[Evolution of serological characteristics in 26 patients with tularemia three years after the outbreak]. / Análisis serológico evolutivo de 26 pacientes con tularemia tres años después del brote.

INTRODUCTION: Tularemia was an unknown zoonosis in our region until the first quarter of 1998. The outbreak that took place on those dates has made possible to study some scantly known characteristics of the disease, as are the determinants for the persistence of a high antibody titer. MATERIAL AND METHODS: Between March and May, 2001, a clinical-analytical control was carried out in 26 patients who showed tularemia in the first quarter of 1998. RESULTS: The clinical forms during the acute phase were: ulceroglandular (46.2% of patients), nodal (26.9%), typhoid (15.4%) and pharyngeal (11.5%). Antibiotics most prescribed were streptomycin and doxycycline. Nine patients showed sequelae three years later. The arithmetical mean of antibody titer declined from 1/1011 in 1998 to 1/187 in 2001. 76.92% of the patients maintained an antibody titer equal or higher than 1/160 three years after the acute phase. A significant relation between the initial antibody titer and the antibody titer registered in the late phase was observed. A relationship between final antibody titer and prescribed antibiotic, presence of sequelae, gender, or age was not demonstrated. CONCLUSIONS: The antibody titer in the presence of Francisella tularensis is high in an important percentage of the patients along years. We should reconsider the standard diagnostic criteria to our community, accepting as a certainty diagnosis exclusively the presence of a demonstrative culture or the existence of seroconversion.