Infection of Nicotiana benthamiana Plants with Potato Virus X (PVX)

12 p.-8 fig.Potato Virus X (PVX) is the type member of Potexvirus genus, a group of plant viruses with a positive-strand RNA genome (~6.4 kb). PVX is able to establish compatible infections in Nicotiana benthamiana, a commonly used host in plant virology, leading to mild symptoms, such as chlorotic mosaic and mottling. PVX has been widely used as a viral vector for more than two decades (Chapman et al., 1992; Baulcombe et al., 1995; Aguilar et al., 2015). It provides a feasible means for the systemic expression in plants of heterologous proteins, such as avirulence factors, proteins with pharmacological properties, etc., (Hammond-Kosack et al., 1995; Gleba et al., 2014), and also as a tool to help decipher the function of genes in plants by virus-induced gene silencing (VIGS) (Lacomme and Chapman, 2008). Two different protocols, i.e., rubbing (A) and agroinfiltration (B), that allow efficient multiplication and propagation of PVX in N. benthamiana are described here in detail. The rubbing method requires previous infected sap, and infection is achieved by inducing mechanical damages to leaf tissue, allowing viral particles to penetrate the plant surface. Agroinfiltration needs previously modified agrobacterium to carry and deliver T-DNA with PVX sequences into the plant cell. Agrobacterium is grown until saturation and infection is established by infiltrating it into plant tissue with a syringe. Any of these two methods can be successfully applied, and the choice should be based mainly on the availability of material and time, but it is recommended to use agroinfiltration when chimeric viruses are being used.This work was supported by the Spanish Ministry of Economy and Competitiveness (Grant Ref. BIO2013-47940-R); The Rural Development Administration (RDA) of the Republic of Korea (Grant Refs. PJ00946102; PJ010246).Peer reviewe

Rethinking Session Variability: Leveraging Session Embeddings for Session Robustness in Speaker Verification

In the field of speaker verification, session or channel variability poses a significant challenge. While many contemporary methods aim to disentangle session information from speaker embeddings, we introduce a novel approach using an additional embedding to represent the session information. This is achieved by training an auxiliary network appended to the speaker embedding extractor which remains fixed in this training process. This results in two similarity scores: one for the speakers information and one for the session information. The latter score acts as a compensator for the former that might be skewed due to session variations. Our extensive experiments demonstrate that session information can be effectively compensated without retraining of the embedding extractor

Striking Deals : Concertation in the Reform of Continental European Welfare States

The reform of the welfare state entails changes in interdependent policy fields stretching from social policies to employment and wage policies. These linked policy fields are often governed by varying sets of corporate actors and involve different decision making procedures. Adaptation in one policy field is often uncoordinated with other policies, and can work at cross-purposes, produce negative externalities, or fail due to missing supporting conditions. The paper has two objectives. It first argues that renewed emergence of tripartite concertation is due to the need to co-ordinate policies across policy fields. Second, it evaluates the institutional factors which have facilitated concertation in some cases, but not in others. Using a similar country design, the paper compares four continental European countries with similar reform pressures but different reform trajectories: France, Germany, Italy, and the Netherlands.Die Reform des Wohlfahrtsstaates erfordert Veränderungen in interdependenten Politikfeldern, von der Sozialpolitik bis hin zur Beschäftigungs- und Lohnpolitik. Diese interdependenten Politikfelder werden von unterschiedlichen Konstellationen korporativer und politischer Akteure kontrolliert und sind unterschiedlichen Verfahren der Entscheidungsfindung unterworfen. Adaptionen in einem Sektor sind häufig nicht mit anderen politischen Entscheidungen koordiniert und können somit negative Auswirkungen haben oder aufgrund der ungünstigen Grundbedingungen fehlschlagen. In dem vorliegenden Discussion Paper wird zunächst argumentiert, daß die Notwendigkeit, politische Entscheidungen über die Grenzen der politischen Sektoren hinaus zu koordinieren, zu einer Renaissance dreiseitiger Konzertierung zwischen Tarifparteien und Regierungen geführt hat. Weiterhin werden die institutionellen Faktoren herausgearbeitet, die eine Konzertierung in einigen Fällen ermöglicht haben, in anderen jedoch nicht. Es werden vier Länder verglichen, die ähnliche Strukturen und Reformzwänge aufweisen, aber unterschiedliche Lösungswege gewählt haben: Frankreich, Deutschland, Italien und die Niederlande

Generation and analysis of large-scale expressed sequence tags (ESTs) from a full-length enriched cDNA library of porcine backfat tissue

BACKGROUND: Genome research in farm animals will expand our basic knowledge of the genetic control of complex traits, and the results will be applied in the livestock industry to improve meat quality and productivity, as well as to reduce the incidence of disease. A combination of quantitative trait locus mapping and microarray analysis is a useful approach to reduce the overall effort needed to identify genes associated with quantitative traits of interest. RESULTS: We constructed a full-length enriched cDNA library from porcine backfat tissue. The estimated average size of the cDNA inserts was 1.7 kb, and the cDNA fullness ratio was 70%. In total, we deposited 16,110 high-quality sequences in the dbEST division of GenBank (accession numbers: DT319652-DT335761). For all the expressed sequence tags (ESTs), approximately 10.9 Mb of porcine sequence were generated with an average length of 674 bp per EST (range: 200–952 bp). Clustering and assembly of these ESTs resulted in a total of 5,008 unique sequences with 1,776 contigs (35.46%) and 3,232 singleton (65.54%) ESTs. From a total of 5,008 unique sequences, 3,154 (62.98%) were similar to other sequences, and 1,854 (37.02%) were identified as having no hit or low identity (<95%) and 60% coverage in The Institute for Genomic Research (TIGR) gene index of Sus scrofa. Gene ontology (GO) annotation of unique sequences showed that approximately 31.7, 32.3, and 30.8% were assigned molecular function, biological process, and cellular component GO terms, respectively. A total of 1,854 putative novel transcripts resulted after comparison and filtering with the TIGR SsGI; these included a large percentage of singletons (80.64%) and a small proportion of contigs (13.36%). CONCLUSION: The sequence data generated in this study will provide valuable information for studying expression profiles using EST-based microarrays and assist in the condensation of current pig TCs into clusters representing longer stretches of cDNA sequences. The isolation of genes expressed in backfat tissue is the first step toward a better understanding of backfat tissue on a genomic basis

A Highly Sensitive Enzyme-Amplified Immunosensor Based on a Nanoporous Niobium Oxide (Nb2O5) Electrode

We report on the development of an enzyme-amplified sandwich-type immunosensor based on a thin gold film sputtered on an anodic nanoporous niobium oxide (Au@Nb2O5) electrode. The electrocatalytic activity of enzymatically amplified electroactive species and a stable electrode consisting of Au@Nb2O5 were used to obtain a powerful signal amplification of the electrochemical immunobiosensor. The method using this electrochemical biosensor based on an Au@Nb2O5 electrode provides a much better performance than those based on conventional bulk gold or niobium oxide electrodes. Our novel approach does not require any time-consuming cleaning steps to yield reproducible electrochemical signals. In addition, the strong adhesion of gold films on the niobium oxide electrodes offers a very stable substrate during electrochemical biosensing. Cyclic voltammetry measurements indicate that non-specific binding of proteins to the modified Au@Nb2O5 surface is sufficiently low to be ignored in the case of our novel system. Finally, we demonstrated the ability of the biosensor based on an Au@Nb2O5 offering the enhanced performance with a high resolution and sensitivity. Therefore, it is expected that the biosensor based on an Au@Nb2O5 has great potential for highly efficient biological devices

Prevalence, Correlates, and Comorbidity of 12-Month Tobacco Dependence among Ever-smokers in South Korea, During 1984-2001

The rate of dependence among ever-users of a drug indicates the risk of developing dependence once an individual has been exposed to the drug. This is the first study to investigate 12-month tobacco dependence (TD) among ever-smokers in a community-based population. Analyses were based on two national studies of representative samples aged 18-64 in 1984 (n=5,025) and in 2001 (n=6,275), conducted with household visits and face-to-face interviews. The rates of 12-month TD among ever-smokers in men showed no significant difference between 51.6% in 1984 and 50.6% in 2001. On the contrary, the rates in women significantly increased from 33.3% in 1984 to 52.8% in 2001. After adjusting for the sociodemographic variables, 'male gender' was significantly associated with 12-month TD among ever-smokers in 1984, but not in 2001. 'Unmarried' was significantly associated in 2001 but not in 1984. 'Alcohol dependence' was the only psychiatric disorder associated with 12-month TD in both study years. In conclusion, 12-month TD was found in about 50% of ever-smokers, and gender differences between the rates of 12-month TD which was observed in 1984 disappeared in 2001. Individuals with 12-month TD showed higher comorbidity with alcohol dependence than ever-smokers without TD

D-PSA-K: A Model for Estimating the Accumulated Potential Damage on Kiwifruit Canes Caused by Bacterial Canker during the Growing and Overwintering Seasons

We developed a model, termed D-PSA-K, to estimate the accumulated potential damage on kiwifruit canes caused by bacterial canker during the growing and overwintering seasons. The model consisted of three parts including estimation of the amount of necrotic lesion in a non-frozen environment, the rate of necrosis increase in a freezing environment during the overwintering season, and the amount of necrotic lesion on kiwifruit canes caused by bacterial canker during the overwintering and growing seasons. We evaluated the model’s accuracy by comparing the observed maximum disease incidence on kiwifruit canes against the damage estimated using weather and disease data collected at Wando during 1994–1997 and at Seogwipo during 2014–2015. For the Hayward cultivar, D-PSA-K estimated the accumulated damage as approximately nine times the observed maximum disease incidence. For the Hort16A cultivar, the accumulated damage estimated by D-PSA-K was high when the observed disease incidence was high. D-PSA-K could assist kiwifruit growers in selecting optimal sites for kiwifruit cultivation and establishing improved production plans by predicting the loss in kiwifruit production due to bacterial canker, using past weather or future climate change data