Viral Strain-Specific Differential Alterations in Arabidopsis Developmental Patterns.

Turnip mosaic virus (TuMV) infections affect many Arabidopsis developmental traits. This paper analyzes, at different levels, the development-related differential alterations induced by different strains of TuMV, represented by isolates UK 1 and JPN 1. The genomic sequence of JPN 1 TuMV isolate revealed highest divergence in the P1 and P3 viral cistrons, upon comparison with the UK 1 sequence. Infectious viral chimeras covering the whole viral genome uncovered the P3 cistron as a major viral determinant of development alterations, excluding the involvement of the PIPO open reading frame. However, constitutive transgenic expression of P3 in Arabidopsis did not induce developmental alterations nor modulate the strong effects induced by the transgenic RNA silencing suppressor HC-Pro from either strain. This highlights the importance of studying viral determinants within the context of actual viral infections. Transcriptomic and interactomic analyses at different stages of plant development revealed large differences in the number of genes affected by the different infections at medium infection times but no significant differences at very early times. Biological functions affected by UK 1 (the most severe strain) included mainly stress response and transport. Most cellular components affected cell-wall transport or metabolism. Hubs in the interactome were affected upon infection.

Lp(a) Levels in Relatives of Patients with Acute Coronary Syndrome and Elevated Lp(a): HER(a) Study.

Background: Lipoprotein(a) [Lp(a)] is a proatherogenic particle associated with increased cardiovascular risk. It is mainly genetically determined; so, the aim of our study is to evaluate the levels of Lp(a) in the relatives of a prospective cohort of patients who have suffered from an acute coronary syndrome (ACS) with Lp(a) ≥ 50 mg/dL. Methods: We conducted a multicenter prospective study, in which consecutive patients who had suffered from an ACS and presented Lp(a) ≥ 50 mg/dL and their first-degree relatives were included. Results: We included 413 subjects, of which 56.4% were relatives of the patients. Family history of early ischemic heart disease was present in 57.5%, and only 20.6% were receiving statin treatment. The family cohort was younger (37.5 vs. 59.1 years; p < 0.001), and 4% had ischemic heart disease and fewer cardiovascular risk factors. Mean Lp(a) levels were 64.9 mg/dL, 59.4% had levels ≥ 50 mg/dL, and 16.1% had levels ≥ 100 mg/dL. When comparing the patients with respect to their relatives, the mean level of Lp(a) was lower but without significant differences regarding the levels of LDLc, ApoB, and non-HDL. However, relatives with Lp(a) ≥ 50 mg/dL, had values similar to the group of patients with ACS (96.8 vs. 103.8 mg/dL; p = 0.18). No differences were found in Lp(a) levels in relatives based on the other lipid parameters. Conclusions: Overall, 59.4% of the first-degree relatives of patients who suffered from an ACS with Lp(a) ≥ 50 mg/dL also had elevated levels. Relatives with elevated Lp(a) had similar levels as patients.

Salicylic acid determines differential senescence produced by two Turnip mosaic virus strains involving reactive oxygen species and early transcriptomic changes.

Losses produced by virus diseases depend mostly on symptom severity. Turnip mosaic virus (TuMV) is one of the most damaging and widespread potyvirus infecting members of the family Brassicaceae, including Arabidopsis thaliana. We used JPN1 and UK1 TuMV strains to characterize viral infections regarding symptom development, senescence progression, antioxidant response, reactive oxygen species (ROS) accumulation, and transcriptional profiling. Both isolates, despite accumulating similar viral titers, induced different symptomatology and strong differences in oxidative status. Early differences in several senescence-associated genes linked to the ORE1 and ORS1 regulatory networks as well as persistent divergence in key ROS production and scavenging systems of the plant were detected. However, at a later stage, both strains induced nutrient competition, indicating that senescence rates are influenced by different mechanisms upon viral infections. Analyses of ORE1 and ORS1 levels in infected Brassica juncea plants showed a similar pattern, suggesting a conserved differential response to both strains in Brassicaceae spp. Transcriptional analysis of the ORE1 and ORS1 regulons showed similarities between salicylic acid (SA) response and the early induction triggered by UK1, the most severe strain. By means of SA-defective NahG transgenic plants, we found that differential senescence progression and ROS accumulation between strains rely on an intact SA pathway.

Cardiovascular prevention in elderly patients.

In recent decades, life expectancy has been increasing significantly. In this scenario, health interventions are necessary to improve prognosis and quality of life of elderly with cardiovascular risk factors and cardiovascular disease. However, the number of elderly patients included in clinical trials is low, thus current clinical practice guidelines do not include specific recommendations. This document aims to review prevention recommendations focused in patients ≥ 75 years with high or very high cardiovascular risk, regarding objectives, medical treatment options and also including physical exercise and their inclusion in cardiac rehabilitation programs. Also, we will show why geriatric syndromes such as frailty, dependence, cognitive impairment, and nutritional status, as well as comorbidities, ought to be considered in this population regarding their important prognostic impact.

Development and evaluation of robust molecular markers linked to disease resistance in tomato for distinctness, uniformity and stability testing.

Molecular markers linked to phenotypically important traits are of great interest especially when traits are difficult and/or costly to be observed. In tomato where a strong focus on resistance breeding has led to the introgression of several resistance genes, resistance traits have become important characteristics in distinctness, uniformity and stability (DUS) testing for Plant Breeders Rights (PBR) applications. Evaluation of disease traits in biological assays is not always straightforward because assays are often influenced by environmental factors, and difficulties in scoring exist. In this study, we describe the development and/or evaluation of molecular marker assays for the Verticillium genes Ve1 and Ve2, the tomato mosaic virus Tm1 (linked marker), the tomato mosaic virus Tm2 and Tm2 ( 2 ) genes, the Meloidogyne incognita Mi1-2 gene, the Fusarium I (linked marker) and I2 loci, which are obligatory traits in PBR testing. The marker assays were evaluated for their robustness in a ring test and then evaluated in a set of varieties. Although in general, results between biological assays and marker assays gave highly correlated results, marker assays showed an advantage over biological tests in that the results were clearer, i.e., homozygote/heterozygote presence of the resistance gene can be detected and heterogeneity in seed lots can be identified readily. Within the UPOV framework for granting of PBR, the markers have the potential to fulfil the requirements needed for implementation in DUS testing of candidate varieties and could complement or may be an alternative to the pathogenesis tests that are carried out at present.

[Tularemia: a decade in the province of Soria (Spain)]. / Tularemia: una década en la provincia de Soria.

INTRODUCTION: Tularemia is a zoonotic disease that has been regularly reported in Spain since 1997. This study analyzes suspected, probable, and confirmed cases of tularemia in the province of Soria, and compares them with tularemia cases recorded in the autonomous community of Castilla y Léon, which, with the exception of 1 sporadic case, occurred in 2 epidemic outbreaks in 1997/1998 and 2007/2008. METHODS: We studied all patients (53) with signs and symptoms of tularemia in the period of 1997 to 2008. Sixty-three serum samples from these patients were tested by a microagglutination assay for antibodies against Francisella tularensis; additionally 10 blood cultures and 1 culture of abscess exudate from an enlarged lymph node were carried out. RESULTS: Over the last decade, 19 cases of tularemia have been diagnosed in Soria (1 sporadic case in 1996, 5 associated with an outbreak reported in 1997/98 and 13 associated with an outbreak occurring in 2007/08). In 95% of the cases, previous contact with hares was reported. The ulceroglandular type was most frequently (62%) observed. F. tularensis was isolated on blood culture in 2 cases. The remaining patients were diagnosed by serology (4 confirmed cases, 13 probable cases). CONCLUSION: The cases of tularemia documented in Soria showed clinical and epidemiological features (predominant ulceroglandular clinical presentation and previous contact with hares) identical to the 1997/98 tularemia outbreak in Castilla y Léon, but contrasted with the 2007/08 outbreak in Castilla y León where typhoidal clinical forms of the disease and a relationship with an increased rodent population (Mycrotus spp) were predominant.

Microbiological diagnosis of catheter-related infections.

This revision describes in detail the different diagnostic techniques of catheter-related infection, both in terms of catheter removal and preservation. Culture techniques based on catheter withdrawal are classified depending on the detection of extraluminal and/or intraluminal colonization, and new methodologies are described. In general, the most important recommendations are: (a) do not send for culture catheter tips without suspicion of infection, (b) Maki's technique is the standard for detecting extraluminal colonization, (c) take 2 pairs of peripheral blood cultures before starting antibiotic treatment, (d) use skin and connections/connectors cultures for the conservative diagnosis due to their high negative predictive value (Gram and culture), and (e) take differential quantitative blood cultures though all catheter lumens and through a peripheral vein.

[Utility of transesophageal echocardiography in young patients with cryptogenic stroke and low cardiovascular risk]. / Utilidad del ecocardiograma transesofágico en pacientes jóvenes con ictus de origen desconocido y bajo riesgo cardiovascular.

BACKGROUND AND OBJECTIVE: Stroke is a high morbimortality disease. In young patients, as many as 40% of acute strokes have no clearly identifiable cause (cryptogenic stroke) and this group of patients had until now limited therapeutic possibilities. However, transesophagical echocardiography (TEE) is changing patient management. PATIENTS AND METHOD: We studied 100 consecutive patients aged 55 years old or less with cryptogenic stroke. TEE was performed in all of them. RESULTS: TEE was normal in 49 patients while in 51 patients it showed any abnormality: patent foramen ovale (PFO) was found in 29 patients, isolated atrial septal aneurysm (ASA) in 1 patient, PFO and ASA in 12 patients, 5 patients had a cardiac mass and in 4 patients we found severe aortic atherosclerotic plaques. Therefore, TEE showed a cardiac source of stroke in 51% of patients. We changed patient management in 46 patients (90.2% of patients with abnormal TEE), indicating percutaneous treatment in 38 patients, surgery in 3 patients and anticoagulant therapy in 5 patients. CONCLUSIONS: TEE found a cardiac cause of stroke in 51% of young patients with cryptogenic stroke. These findings led to change the therapeutic management in 47% of patients. TEE seems to be a useful technique in young patients with cryptogenic stroke.

A developmentally linked, dramatic, and transient loss of virus from roots of Arabidopsis thaliana plants infected by either of two RNA viruses.

Possible effects of host developmental stage on the amount of virus present in systemically infected plant tissues hitherto have received little attention. In this study, the pattern of virus accumulation over the plant lifespan has been examined in systemically invaded tissues of Arabidopsis thaliana infected by either of two distinct (+)RNA viruses: Turnip mosaic virus, a member of Potyvirus, and Oilseed rape mosaic virus, a member of Tobamovirus. Quantitative analyses of virus coat protein and virus genomic RNA in roots versus aerial plant parts revealed generally sinusoidal temporal patterns of virus accumulation. In noninoculated leaves, a time period was found during which no virus accumulation was detected. This period was coincident with the approximately 7 days of inflorescence bud formation and differentiation. In roots, virion content reached high levels a few days after inoculation, dropping dramatically during the period of bud formation and quickly recovering after it. These results, together with electron microscopy observations, are consistent with loss of virions due to disassembly. Fluorescence observations of green fluorescent protein-tagged virus-infected root tissue also were consistent with a net loss of virus-specified proteins. Inoculations performed after the emergence of the inflorescence and on A. thaliana flowering-time mutants support the temporal link between observed changes in virus content and inflorescence bud formation. Different host-involving biochemical processes can be invoked to provide mechanistic clues, but no one of them alone seems sufficient to explain the complex patterns of tight temporal regulation of virus accumulation observed in these experiments.

Nanonets Derived from Turnip Mosaic Virus as Scaffolds for Increased Enzymatic Activity of Immobilized Candida antarctica Lipase B.

Elongated flexuous plant viral nanoparticles (VNPs) represent an interesting platform for developing different applications in nanobiotechnology. In the case of potyviruses, the virion external surface is made up of helically arrayed domains of the viral structural coat protein (CP), repeated over 2000 times, in which the N- and C-terminal domains of each CP are projected toward the exterior of the external virion surface. These characteristics provide a chemical environment rich in functional groups susceptible to chemical conjugations. We have conjugated Candida antarctica lipase B (CALB) onto amino groups of the external surface of the potyvirus turnip mosaic virus (TuMV) using glutaraldehyde as a conjugating agent. Using this approach, TuMV virions were transformed into scaffolds for CALB nanoimmobilization. Analysis of the resulting structures revealed the formation of TuMV nanonets onto which large CALB aggregates were deposited. The functional enzymatic characterization of the CALB-bearing TuMV nanonets showed that CALB continued to be active in the nanoimmobilized form, even gaining an increased relative specific activity, as compared to the non-immobilized form. These novel virus-based nanostructures may provide a useful new approach to enzyme nanoimmobilization susceptible to be industrially exploited.

Ultra-sensitive detection of two garlic potyviruses using a real-time fluorescent (Taqman) RT-PCR assay.

A method for the detection of Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), the two most prevalent garlic potyviruses, has been developed that combines IC-RT-PCR/RT-PCR with the use of TaqMan probes. Comparisons with ELISA results obtained with identical OYDV and LYSV infected samples showed sensitivity in detecting these viruses increased up to 10(6)-fold. OYDV and LYSV were detected using different fluorochromes in the probe, thus allowing unequivocal diagnosis for each of them. The polyvalence of the designed virus-specific primers and probes was shown through their application to the detection of three isolates from very different geographical areas and from different hosts. A second version of the method avoids the need for an immunocapture step through the performance of a TaqMan RT-PCR assay directly on extracts of garlic cloves. This modification on the proposed basic method allows the analysis of bulb samples in 3-4h but did not give reproducible results with leaves. Both versions of the new diagnostic method bear great potential for their implementation in virus-free certification schemes in garlic, a vegetatively propagated crop for which such a certification is critical for a high-quality product.

Microbiological diagnosis of catheter-related infections / Diagnóstico microbiológico de las infecciones asociadas a catéteres intravasculares

This revision describes in detail the different diagnostic techniques of catheter-related infection, both in terms of catheter removal and preservation. Culture techniques based on catheter withdrawal are classified depending on the detection of extraluminal and/or intraluminal colonization, and new methodologies are described. In general, the most important recommendations are: (a) do not send for culture catheter tips without suspicion of infection, (b) Maki's technique is the standard for detecting extraluminal colonization, (c) take 2 pairs of peripheral blood cultures before starting antibiotic treatment, (d) use skin and connections/connectors cultures for the conservative diagnosis due to their high negative predictive value (Gram and culture), and (e) take differential quantitative blood cultures though all catheter lumens and through a peripheral vein

Esta revisión describe con detalle las diferentes técnicas diagnósticas de infección relacionada con el catéter, tanto con la retirada como con la conservación del mismo. Las técnicas de cultivo basadas en la retirada del catéter se clasifican en base a la detección de colonización extraluminal, intraluminal, o ambas, asimismo, se describen nuevas metodologías. De forma general, las recomendaciones más importantes son: a) no enviar para cultivo puntas de catéter retiradas sin sospecha de infección, b) la técnica de Maki es el estándar válido de detección de colonización extraluminal, c) tomar 2 parejas de hemocultivos de sangre periférica antes de iniciar tratamiento antibiótico, d) utilizar cultivos de piel y conexiones/conectores para el diagnóstico conservador por su alto valor predictivo negativo (Gram y cultivo), y e) extraer hemocultivos cuantitativos diferenciales por todas las luces del catéter y por vena periférica

Diagnóstico microbiológico de la tuberculosis. 20 años en la provincia de Soria. Rendimiento y oportunidades básicas de mejora / (Microbiological diagnosis of tuberculosis. 20 years in the province of Soria. Performance and basic opportunities of improvement

Introducción. El objetivo del estudio fue conocer los datos relativos al diagnóstico microbiológico de la tuberculosis en la provincia de Soria, así como analizar la rentabilidad diagnóstica de las técnicas utilizadas y la utilización del laboratorio de microbiología en lo que concierne a la tuberculosis. Métodos. Se diseñó un estudio observacional, descriptivo y retrospectivo, incluyendo todos los pacientes con tuberculosis de cualquier localización que tuviesen su residencia en la provincia de Soria. El periodo de estudio abarcó los casos diagnosticados entre 1994 y 2013 realizando seguimiento durante 24 meses tras el inicio del tratamiento. Resultados. Se detectaron 337 pacientes durante el periodo estudiado. En más del 3% de los pacientes no se envió ninguna muestra al laboratorio de microbiología, porcentaje que ascendió al 23% en tuberculosis osteoarticulares y 33% en tuberculosis linfáticas. Se obtuvo confirmación microbiológica en el 80% y la baciloscopia fue positiva en el 32%. Las muestras se sembraron en medios sólidos y líquidos; el 10% de las cepas sólo se aislaron en un tipo de medio. El porcentaje de cepas resistentes a isoniazida fue de 2,9%, se detectó una cepa multirresistente (0,3%) y una cepa con resistencia únicamente a rifampicina (0,3%). De todos los pacientes bacilíferos con seguimiento, no se envió ninguna muestra para estudiar la negativización en el 36%. Conclusión. Destaca la necesidad de mantener el cultivo combinado en medios líquidos y sólidos. Es necesario potenciar el uso del laboratorio de microbiología, enviando todas las posibles muestras diagnósticas y realizando controles bacteriológicos de seguimiento para objetivar la curación

Introduction. The aim of the study was to describe the bacteriological diagnosis of the tuberculosis in the province of Soria (Spain), as well as to analyse the techniques diagnostic performance and the use of the microbiology laboratory regarding tuberculosis. Methods. An observational, descriptive and retrospective study was designed, including all patients with tuberculosis of any location that had their residence in the province of Soria. The period of study included patients diagnosed between 1994 and 2013 and a 24 months follow-up after the beginning of treatment was realized. Results. A total of 337 patients were detected during the studied period. No sample was sent to the microbiology laboratory in more than 3% of the patients (23% in skeletal tuberculosis and 33% in lymphatic tuberculosis). Bacteriological confirmation was obtained in 80% and 32% were smear-positive. Specimens were culture on solid and in liquid media; 10% of the strains were only isolated in one type of media. There were 2.9% isoniazid-resistant strains, 0.3% multi-drug resistant strains, and 0.3% rifampicin-resistant strains. A total of 36% of smear-positive pulmonary tuberculosis patients had no specimens sent for a follow-up study. Conclusion. It is essential to combine the use of a liquid and a solid medium. Physicians should be encouraged to submit specimens for mycobacteriological diagnostic and follow-up

Fiabilidad del conjunto mínimo básico de datos como herramienta epidemiológica de la enfermedad tuberculosa / Reliability of the Minimum Basic Data Set as an Epidemiological Tool in Tuberculosis

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[Improvement in functional capacity after levothyroxine treatment in patients with chronic heart failure and subclinical hypothyroidism]. / Mejoría de la capacidad funcional tras tratamiento con levotiroxina en pacientes con insuficiencia cardíaca crónica e hipotiroidismo subclínico.

AIM: To assess whether levothyroxine treatment improves functional capacity in patients with chronic heart failure (New York Heart Association class i-iii) and subclinical hypothyroidism. METHODS: One hundred and sixty-three outpatients with stable chronic heart failure followed up for at least 6 months were enrolled. A physical examination was performed, and laboratory tests including thyroid hormone levels, Doppler echocardiogram, radionuclide ventriculography, and Holter monitoring were requested. Functional capacity was assessed by of the 6-min walk test. Patients with subclinical hypothyroidism were detected and, after undergoing the s6-min walk test, were given replacement therapy. When they reached normal thyrotropin (TSH) levels, the 6-min walk test was performed again. The distance walked in both tests was recorded, and the difference in meters covered by each patient was analyzed. RESULTS: Prevalence of subclinical hypothyroidism in patients with heart failure was 13%. These patients walked 292±63m while they were hypothyroid and 350±76m when TSH levels returned to normal, a difference of 58±11m (P<.011). Patients with normal baseline TSH levels showed no significant difference between the 2 6-min walk tests. CONCLUSIONS: Patients with chronic heart failure and subclinical hypothyroidism significantly improved their physical performance when normal TSH levels were reached.

Evaluation of a fluorogenic real-time reverse transcription-polymerase chain reaction method for the specific detection of all known serotypes of porcine teschoviruses.

Performance of a real-time reverse-transcription polymerase chain reaction method for the rapid, simple and reliable detection of porcine teschovirus (PTV) was assessed. The method was based on the use of a set of oligonucleotides consisting of two specific primers and a fluorogenic TaqMan-MGB probe. Reverse transcription and PCR reactions were performed sequentially in one step. As a result the whole procedure was simple and rapid, taking less than 3h for completion. The method reacted in a dose-dependent manner with prototype strains for the eleven known PTV serotypes (PTV1-11), with higher analytical sensitivity than other gel-based RT-PCR methods described, which were performed in parallel to allow for a comparison. The assay did not cross-react with other related viruses or porcine viruses tested. The diagnostic performance of the method was analyzed using a panel of field samples consisting of pig fecal and pig slurry samples. As a conclusion, this technique is adequate and convenient for porcine teschovirus detection, both for diagnosis as well as in environmental investigations.