Quantitative detection of _Potato virus Y_ in potato plants and aphids - Discussion of diverse applications in potato research

Every year potato growers worldwide complain about severe yield losses caused by _Potato virus Y_ (PVY). Therefore, PVY along with _Potato leafroll virus_ belongs to the most important potato viruses. There are three main strains of PVY: PVY^O^, PVY^N^ and PVY^C^. However, also recombinant forms exist such as PVY^N^Wilga and PVY^NTN^, both of which increase in importance due to their potential to displace the non-recombinant strains at a high percentage. They appear also in mixed infections. In recent years PCR and qPCR assays were developed to differentiate PVY isolates. In order to identify PVY isolates by PCR often large amplicons have to be generated which requires the input of expensive enzymes. On the other hand, qPCR assays until now do not allow the differentiation between PVY^N^Wilga and PVY^NTN^. 

For the discrimination between PVY^O^/PVY^N^Wilga and PVY^N^/PVY^NTN^ a qPCR assay was developed, which allows the differentiation and highly efficient quantification of both strains and recombinants, respectively. For this purpose dual-labeled hydrolysis probes tagged with different fluorophores were designed. The assay is suitable for many different applications, for example safety research on genetically modified (GM) potato plants. The goal of this research is to determine whether genetic modification causes changes in resistance to viruses. Two different GM cultivars were examined for signs of altered resistance to an infection with PVY in comparison to their near-isogenic lines and three reference cultivars. Reference cultivars are included to determine the baselines for resistance and thus to be able to decide if the changes could represent a biological risk. The plants to be investigated were mechanically inoculated with PVY^N^Wilga or PVY^NTN^ and analyzed by means of the developed assay after two weeks. The results of the experiment indicate that the differences in virus titer between the reference cultivars are higher than between the GM potatoes and their isogenic lines. Therefore, in our experiments the GM potato plants showed no alteration in PVY resistance to neither one of the tested strains.

Since _Myzus persicae_ is one of the most important vectors transmitting PVY, the developed assay will also be applied to the quantification of PVY particles in aphids. The displacement of PVY^O^ and PVY^N^ by PVY^N^Wilga and PVY^NTN^ may be due to a difference in efficiency of transmission by _M. persicae_. Therefore, the objective is to test whether more virus particles of the recombinant forms in comparison to the non-recombinant strains PVY^O^ and PVY^N^ bind in the stylets of _M. persicae_. 

A third possible application of the developed assay may be of interest in potato breeding. The exact quantification of PVY particles in plants allows the classification of resistance in potato plants. It is possible to estimate whether a resistance is extreme or not. Extreme resistance is characterized by the absence or presence of very low amounts of virus particles in plants several days after inoculation. When testing the plants for PVY infection by ELISA, often unspecific reactions occur which makes it difficult to differentiate between plants weakly infected and plants very weakly infected. An exact quantification of the PVY titer gives more certainty for the determination of the resistance type.

In conclusion, the developed assay is an efficient and low-cost method that allows the differentiation and quantification of PVY^O^/PVY^N^Wilga on the one hand and PVY^N^/PVY^NTN^ on the other hand with high throughput. The method can be utilized for a wide range of applications in potato research.
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THE ASSESMENT OF COGNITIVE IMPAIRMENT FOR STROKE PATIENT

Introduction: Stroke is a leading cause of long-term disability. Deficit neurological of stroke have impact in physical, emotional, social, and cognitive aspects. The impact of deficit neurological may affects long-term quality of life. The purpose of this study was to evaluate the effectiveness of cognitive impairment assessment for stroke. Method: Using an electronic database including Pubmed, Sciencedirect, Medline Ebsco Host, Proquest and Springerlink Library only for English language articles. Combining cognitive impairment, screening cognitive, screening MoCA, MoCA tools, screening MMSE, MMSE tools, and stroke as the search keywords. The 603 articles retrieved. Using matching keywords, 40 articles were selected. Finally, 4 articles were selected which are study of test accuracy among consecutive patients. Result: Using MoCA test more effective to evaluate cognitive impairment for stroke patients. Discussion: The MoCA test for patient post stroke can detect cognitive impairment compared using the MMSE. The MMSE subtest domain score can’t distinguished, while the MoCA subtest domain score (visuospatial/executive function, attention and recall) can be distinguished. Assessing executive function can help determine a patient’s capacity to execute health care and discharge planning decisions. Keywords: MoCA, MMSE, strok

Giant spin-orbit effects on H-1 and C-13 NMR shifts for uranium(VI) complexes revisited: role of the exchange-correlation response kernel, bonding analyses, and new predictions

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.Previous relativistic quantum-chemical predictions of unusually large H-1 and C-13 NMR chemical shifts for ligand atoms directly bonded to a diamagnetic uranium(VI) center (P. Hrobarik, V. Hrobarikova, A. H. Greif and M. Kaupp, Angew. Chem., Int. Ed., 2012, 51, 10884) have been revisited by two- and four-component relativistic density functional methods. In particular, the effect of the exchange-correlation response kernel, which had been missing in the previously used two-component version of the Amsterdam Density Functional program, has been examined. Kernel contributions are large for cases with large spin-orbit (SO) contributions to the NMR shifts and may amount to up to similar to 30% of the total shifts, which means more than a 50 ppm difference for the metal-bonded carbon shifts in some extreme cases. Previous calculations with a PBE-40HF functional had provided overall reasonable predictions, due to cancellation of errors between the missing kernel contributions and the enhanced exact-exchange (EXX) admixture of 40%. In the presence of an exchange-correlation kernel, functionals with lower EXX admixtures give already good agreement with experiments, and the PBE0 functional provides reasonable predictive quality. Most importantly, the revised approach still predicts unprecedented giant H-1 NMR shifts between +30 ppm and more than +200 ppm for uranium(VI) hydride species. We also predict uranium-bonded C-13 NMR shifts for some synthetically known organometallic U(VI) complexes, for which no corresponding signals have been detected to date. In several cases, the experimental lack of these signals may be attributed to unexpected spectral regions in which some of the C-13 NMR shifts can appear, sometimes beyond the usual measurement area. An extremely large uranium-bonded C-13 shift above 550 ppm, near the upper end of the diamagnetic C-13 shift range, is predicted for a known pincer carbene complex. Bonding analyses allow in particular the magnitude of the SO shifts, and of their dependence on the functional, on the ligand position in the complex, and on the overall electronic structure to be better appreciated, and improved confidence ranges for predicted shifts have been obtained

Distinct transthyretin oxidation isoform profile in spinal fluid from patients with Alzheimer’s disease and mild cognitive impairment

BACKGROUND: Transthyretin (TTR), an abundant protein in cerebrospinal fluid (CSF), contains a free, oxidation-prone cysteine residue that gives rise to TTR isoforms. These isoforms may reflect conditions in vivo. Since increased oxidative stress has been linked to neurodegenerative disorders such as Alzheimer’s disease (AD) it is of interest to characterize CSF-TTR isoform distribution in AD patients and controls. Here, TTR isoforms are profiled directly from CSF by an optimized immunoaffinity-mass spectrometry method in 76 samples from patients with AD (n = 37), mild cognitive impairment (MCI, n = 17)), and normal pressure hydrocephalus (NPH, n = 15), as well as healthy controls (HC, n = 7). Fractions of three specific oxidative modifications (S-cysteinylation, S-cysteinylglycinylation, and S-glutathionylation) were quantitated relative to the total TTR protein. Results were correlated with diagnostic information and with levels of CSF AD biomarkers tau, phosphorylated tau, and amyloid β(1-42) peptide. RESULTS: Preliminary data highlighted the high risk of artifactual TTR modification due to ex vivo oxidation and thus the samples for this study were all collected using strict and uniform guidelines. The results show that TTR is significantly more modified on Cys(10) in the AD and MCI groups than in controls (NPH and HC) (p ≤ 0.0012). Furthermore, the NPH group, while having normal TTR isoform distribution, had significantly decreased amyloid β peptide but normal tau values. No obvious correlations between levels of routine CSF biomarkers for AD and the degree of TTR modification were found. CONCLUSIONS: AD and MCI patients display a significantly higher fraction of oxidatively modified TTR in CSF than the control groups of NPH patients and HC. Quantitation of CSF-TTR isoforms thus may provide diagnostic information in patients with dementia symptoms but this should be explored in larger studies including prospective studies of MCI patients. The development of methods for simple, robust, and reproducible inhibition of in vitro oxidation during CSF sampling and sample handling is highly warranted. In addition to the diagnostic information the possibility of using TTR as a CSF oxymeter is of potential value in studies monitoring disease activity and developing new drugs for neurodegenerative diseases

When ESDP confronts ENP : security sector reform as a bridge in the EU's foreign policy

The European Neighbourhood Policy's birth has taken place in parallel with the renewed momentum of the European Security and Defence Policy, which has launched 14 operations since 2003. Both policies' instruments have converged in the neighbouring area covered by ENP: Georgia, in the East and the Palestinian Territories in the South. In both cases, the Security Sector Reform strategies have been the main focus for ESDP and an important objective for ENP. In this paper, two objectives are pursued: first, to assess the EU's involvement in both cases in SSR terms; and second, to analyse whether the convergence of ESDP operations with a broader EU neighbourhood policy implies that the former has become an instrument for the a EU external action

Ambient air particulate total lung deposited surface area (LDSA) levels in urban Europe

This study is supported by the RI-URBANS project (Research Infrastructures Services Reinforcing Air Quality Monitoring Capacities in European Urban & amp; Industrial Areas, European Union's Horizon 2020 research and innovation program, Green Deal, European Commission, contract 101036245). This study is also supported by National Natural Science Foundation of China (42101470, 72242106) and in part by the Chunhui Project Foundation of the Education Department of China under Grant HZKY20220053. This study benefited from the Aerosol, Clouds and Trace Gases Research Infrastructure (ACTRIS), especially the so-called ACTRIS-2 H2020 research project (grant no 654109), and the authors would like to thank ACTRIS (The Aerosol, Clouds and Trace Gases Research Infrastructure), especially the ACTRIS in situ EBAS Data Centre (EBAS), for providing datasets to the study. This study is also partly funded by the National Institute for Health Research (NIHR) Health Protection Research Unit in Environmental Exposures and Health, a partnership between UK Health Security Agency (UKHSA) and Imperial College London, and the UK Natural Environment Re-search Council, and the views expressed are those of the author(s) and not necessarily those of the NIHR, UKHSA or the Department of Health and Social Care. The research was also supported by the Hungarian Research, Development and Innovation Office (grant no. K132254). We thank also the support from "Agencia Estatal de Investigacion" from the Spanish Ministry of Science and Innovation, and FEDER funds under the projects CAIAC (PID2019-108990RB-I00); and the Generalitat de Catalunya (AGAUR 2017 SGR41) and the Direccio General de Territori. IMT Nord Europe and LOA acknowledge financial support from the Labex CaPPA project, funded by the French National Research Agency (ANR-11-LABX-0005-01), and the CLIMIBIO and ECRIN projects, both financed by the Regional Council "Hauts-de-France" and the European Regional Development Fund (ERDF).This study aims to picture the phenomenology of urban ambient total lung deposited surface area (LDSA) (including head/throat (HA), tracheobronchial (TB), and alveolar (ALV) regions) based on multiple path particle dosimetry (MPPD) model during 2017-2019 period collected from urban background (UB, n = 15), traffic (TR, n = 6), suburban background (SUB, n = 4), and regional background (RB, n = 1) monitoring sites in Europe (25) and USA (1). Briefly, the spatial-temporal distribution characteristics of the deposition of LDSA, including diel, weekly, and seasonal pat-terns, were analyzed. Then, the relationship between LDSA and other air quality metrics at each monitoring site was investigated. The result showed that the peak concentrations of LDSA at UB and TR sites are commonly observed in the morning (06:00-8:00 UTC) and late evening (19:00-22:00 UTC), coinciding with traffic rush hours, biomass burning, and atmospheric stagnation periods. The only LDSA night-time peaks are observed on weekends. Due to the variability of emission sources and meteorology, the seasonal variability of the LDSA concentration revealed sig-nificant differences (p = 0.01) between the four seasons at all monitoring sites. Meanwhile, the correlations of LDSA with other pollutant metrics suggested that Aitken and accumulation mode particles play a significant role in the total LDSA concentration. The results also indicated that the main proportion of total LDSA is attributed to the ALV fraction (50 %), followed by the TB (34 %) and HA (16 %). Overall, this study provides valuable information of LDSA as a predictor in epidemiological studies and for the first time presenting total LDSA in a variety of European urban environments.RI-URBANS project (Research Infrastructures Services Reinforcing Air Quality Monitoring Capacities in European Urban amp; Industrial Areas, European Union's Horizon 2020 research and innovation program, Green Deal, European Commission, 101036245)National Natural Science Foundation of China (NSFC)Chunhui Project Foundation of the Education Department of ChinaAerosol, Clouds and Trace Gases Research Infrastructure (ACTRIS)National Institute for Health Research (NIHR) Health Protection Research Unit in Environmental Exposures and HealthUK Research & Innovation (UKRI) Natural Environment Research Council (NERC)National Research, Development & Innovation Office (NRDIO) - Hungary"Agencia Estatal de Investigacion" from the Spanish Ministry of Science and InnovationGeneralitat de Catalunya 42101470, 72242106Direccio General de Territori HZKY20220053Agence Nationale de la Recherche (ANR) 654109Regional Council "Hauts-de-France"European Union (EU) K132254, PID2019-108990RB-I00, AGAUR 2017 SGR41, ANR-11-LABX-0005-01ERD

Spatial and temporal variability of urban fluxes of methane, carbon monoxide and carbon dioxide above London, UK

We report on more than 3 years of measurements of fluxes of methane (CH4), carbon monoxide (CO) and carbon dioxide (CO2) taken by eddy-covariance in central London, UK. Mean annual emissions of CO2 in the period 2012–2014 (39.1 ± 2.4 ktons km−2 yr−1) and CO (89 ± 16 tons km−2 yr−1 ) were consistent (within 1 and 5% respectively) with values from the London Atmospheric Emissions Inventory, but measured CH4 emissions (72 ± 3 tons km−2 yr−1) were over two-fold larger than the inventory value. Seasonal variability was large for CO with a winter to summer reduction of 69 %, and monthly fluxes were strongly anti-correlated with mean air temperature. The winter increment in CO emissions was attributed mainly to vehicle cold starts and reduced fuel combustion efficiency. CO2 fluxes were 33 % higher in winter than in summer and anti-correlated with mean air temperature, albeit to a lesser extent than for CO. This was attributed to an increased demand for natural gas for heating during the winter. CH4 fluxes exhibited moderate seasonality (21 % larger in winter), and a spatially variable linear anti-correlation with air temperature. Differences in resident population within the flux footprint explained up to 90 % of the spatial variability of the annual CO2 fluxes and up to 99 % for CH4. Furthermore, we suggest that biogenic sources of CH4, such as wastewater, which is unaccounted for by the atmospheric emissions inventories, make a substantial contribution to the overall bud- get and that commuting dynamics in and out of central business districts could explain some of the spatial and temporal variability of CO2 and CH4 emissions. To our knowledge,this study is unique given the length of the data sets presented, especially for CO and CH4 fluxes. This study offers an independent assessment of “bottom-up” emissions inventories and demonstrates that the urban sources of CO and CO2 are well characterized in London. This is however not the case for CH4 emissions which are heavily underestimated by the inventory approach. Our results and others point to opportunities in the UK and abroad to identify and quantify the “missing” sources of urban methane, revise the methodologies of the emission inventories and devise emission reduction strategies for this potent greenhouse gas