The complete genome sequence of a divergent grapevine virus I isolate naturally infecting grapevine in Greece.

A significant number of new members of the genus Vitivirus have been identified recently, mainly due to the advent of high-throughput sequencing (HTS). Grapevine virus I (GVI), which was identified in New Zealand in 2018, is one of these viruses. RNAseq HTS analysis of a Greek grapevine (cv. Daphnia), revealed the presence of a GVI-like isolate (D2-1/19). Sequence analysis confirmed the classification of D2-1/19 as GVI. However, both sequence and phylogenetic data exhibited high levels of variability between D2-1/19 and the previously characterized GVI isolates. This study provides the full-length sequence of a divergent GVI isolate, adding knowledge to the limited information available about this recently identified virus.

Flexible Passive near Field Communication Tag for Multigas Sensing.

In this work we present a full-passive flexible multigas sensing tag for the determination of oxygen, carbon dioxide, ammonia, and relative humidity readable by a smartphone. This tag is based on near field communication (NFC) technology for energy harvesting and data transmission to a smartphone. The gas sensors show an optic response that is read through high-resolution digital color detectors. A white LED is used as the common optical excitation source for all the sensors. Only a reduced electronics with very low power consumption is required for the reading of the optical responses and data transmission to a remote user. An application for the Android operating system has been developed for the power supplying and data reception from the tag. The responses of the sensors have been calibrated and fitted to simple functions, allowing a fast prediction of the gases concentration. Cross-sensitivity has also been evaluated, finding that in most of the cases it is negligible or easily correctable using the rest of the readings. The election of the target gases has been due to their importance in the monitoring of modified atmosphere packaging. The resolutions and limits of detection measured are suitable for such kinds of applications.

MALDI-TOF mass spectrometry as a tool for the discrimination of high-risk Escherichia coli clones from phylogenetic groups B2 (ST131) and D (ST69, ST405, ST393).

Reliable, quick and low-cost methods are needed for the early detection of multidrug-resistant and highly virulent high-risk B2 and D Escherichia coli clones or clonal complexes (HiRCC). Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) seems to have a good discriminatory potential at different subspecies levels, but it was never evaluated for the discrimination of E. coli clones. We assessed the potential of MALDI-TOF MS coupled to multivariate data analysis to discriminate representative E. coli B2 and D HiRCC. Seventy-three E. coli isolates from B2 (including ST131 and B2 non-ST131 clones) and D (ST69, ST393, ST405) with variable pulsed-field gel electrophoresis (PFGE) patterns, origins and dates (1980-2010) were tested. MS spectra were acquired from independent extracts obtained from different plate cultures in two different Microflex LT MALDI-TOF devices (Bruker) after a standard extraction procedure. MALDI-TOF MS fingerprinting analysis revealed a good discriminatory ability between the four HiRCC analysed (ST131, ST69, ST405, ST393) and between B2 ST131 and other B2 non-ST131 isolates. Clusters defined by MALDI-TOF MS were consistent with the clonal complexes assigned by multilocus sequence typing (MLST), although differences were detected regarding the composition of clusters obtained by the comparison of PFGE profiles. We demonstrate, for the first time, that characteristic mass fingerprints of different E. coli HiRCC are sufficiently discriminatory and robust to enable their differentiation by MALDI-TOF MS, which might represent a promising tool for the optimisation of infection control, individual patient management and large-scale epidemiological studies of public health relevance. The good correlation between phenotypic and genotypic features further corroborates phylogenetic relationships delineated by MLST.

Modelling of salt uptake for salt content standardization in dry-cured ham.

Salt content standardization in dry-cured ham production is complex as there are many factors that influence salt uptake. The aim of this work was to study and model salt uptake as a function of salting time and the characteristics of green ham in two different ham types using a large amount of data obtained from an inline non-invasive characterization technology. The usefulness of these models to standardize the salt content of the industrial production was also evaluated. Magnetic induction technology was used to characterize lean and fatty hams in order to further model the salt uptake using different statistical methods. Salt content increased with the salting time showing an asymptotic behaviour which was lower in fatty hams. The best models were those that considered the salting time, weight, and fat content of hams as model independent variables. These models are helpful for salt content standardization within the industry and can be used to simulate process modifications and avoid the time-consuming and costly trial and error tests.

Liver transplantation outcome in patients with angiographically proven coronary artery disease: a multi-institutional study.

Over the last decade the age of liver transplant (LT) recipients and the likelihood of coronary artery disease (CAD) in this population have increased. There are no multicenter studies that have examined the impact of CAD on LT outcomes. In this historical cohort study, we identified adult LT recipients who underwent angiography prior to transplantation at seven institutions over a 12-year period. For each patient we recorded demographic data, recipient and donor risk factors, duration of follow-up, the presence of angiographically proven obstructive CAD (≥50% stenosis) and post-LT survival. Obstructive CAD was present in 151 of 630 patients, the CAD(+) group. Nonobstructive CAD was found in 479 patients, the CAD(-) group. Patient survival was similar for the CAD(+) group (adjusted HR 1.13, CI = [0.79, 1.62], p = 0.493) compared to the CAD(-) group. The CAD(+) patients were further stratified into severe (CADsev, >70% stenosis, n = 96), and moderate CAD (CADmod, 50-70% stenosis, n = 55) groups. Survival for the CADsev (adjusted HR = 1.26, CI = [0.83, 1.91], p = 0.277) and CADmod (adjusted HR = 0.93, CI = [0.52, 1.66], p = 0.797) groups were similar to the CAD(-) group. We conclude that when current CAD treatment strategies are employed prior to transplant, post-LT survival is not significantly different between patients with and without obstructive CAD.

Screen printed flexible radiofrequency identification tag for oxygen monitoring.

In this work, a radiofrequency identification (RFID) tag with an optical indicator for the measurement of gaseous oxygen is described. It consists of an O2 sensing membrane of PtOEP together with a full electronic system for RFID communication, all printed on a flexible substrate. The membrane is excited by an LED at 385 nm wavelength and the intensity of the luminescence generated is registered by means of a digital color detector. The output data corresponding to the red coordinate of the RGB color space is directly related to the concentration of O2, and it is sent to a microcontroller. The RFID tag is designed and implemented by screen printing on a flexible substrate for the wireless transmission of the measurement to a remote reader. It can operate in both active and passive mode, obtaining the power supply from the electromagnetic waves of the RFID reader or from a small battery, respectively. This system has been fully characterized and calibrated including temperature drifts, showing a high-resolution performance that allows measurement of very low values of oxygen content. Therefore this system is perfectly suitable for its use in modified atmosphere packaging where the oxygen concentration is reduced below 2%. As the reading of the O2 concentration inside the envelope is carried out with an external RFID reader using wireless communication, there is no need for perforations for probes or wires, so the packaging remains completely closed. With the presented device, a limit of detection of 40 ppm and a resolution as low as 0.1 ppm of O2 can be reached with a low power consumption of 3.55 mA.

Non-diphtheriae Corynebacterium species: an emerging respiratory pathogen.

The purpose of the study was to describe the microbiological and clinical features of ten cases of lower respiratory tract infection due to Corynebacterium striatum, Corynebacterium propinquum and Corynebacterium pseudodiphtheriticum. Respiratory samples were recovered from hospitalised patients who were diagnosed of pneumonia and exacerbations of chronic respiratory infections. The samples were Gram-stained and seeded on conventional bacterial growing media. Bacteria were identified by matrix-assisted linear desorption/ionisation-time-of-flight mass spectrometry (MALDI-TOF MS). Antibiotic susceptibility was tested by the disk diffusion method. All patients presented an acute respiratory onset, most of them in the context of an underlying disease and/or immunosuppression. In all patients, the microscopical examination of Gram-stained respiratory samples showed numerous polymorphonuclear cells and Gram-positive bacilli, suggestive of the Corynebacterium morphotype. A pure culture growth of Corynebacterium was obtained in the majority (72 %) of samples. The conclusions are that non-diphtheriae Corynebacterium species are an emerging cause of respiratory infection among patients with chronic respiratory disease and/or immunosuppression, and cannot always be considered as mere colonisers. The microorganism's predominance in Gram-stained purulent respiratory samples together with abundant growth in the culture is the key for the microbiological diagnosis.

Study of the first step of hydrogen release in ammonia borane using high-resolution Raman spectroscopy and different heating ramps.

Ammonia borane, as a source of hydrogen, has attracted much attention due to its high hydrogen content, low molecular weight, and high stability in solution. However, the process and enhancement of hydrogen release must be done practically under ambient conditions. For this work, Raman spectroscopy, principal component analysis (PCA), and molecular simulation were applied to study the hydrogen release process of ammonia borane. Three stages of release were observed from room temperature to 1300 °C. The shift, the appearance, and the disappearance of the Raman bands were evident in the whole process. In-situ monitoring of Raman and PCA, with four different heating rates between 70 and 130 °C, was done; ammonia borane showed visible variations in its first release step, in which a fast rate helped reduce distortion in the release process. Finally, molecular simulation of ammonia borane using the Density Functional Theory (DFT) in calculations showed that dihedral rotation and stretching of the hydrogen bonds can occur before the first release step.

Clinical and microbiological features of a cystic fibrosis patient chronically colonized with Pandoraea sputorum identified by combining 16S rRNA sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry.

Clonal isolates identified as various nonfermentative Gram-negative bacilli over a 5-year period from sputum cultures of a 30-year-old cystic fibrosis patient were successfully reidentified as Pandoraea sputorum by combining 16S rRNA sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Decreased lung function improved after 1 year of azithromycin and inhaled 7%-hypertonic saline treatment.

Smart facemask for wireless CO2 monitoring.

The use of facemasks by the general population is recommended worldwide to prevent the spread of SARS-CoV-2. Despite the evidence in favour of facemasks to reduce community transmission, there is also agreement on the potential adverse effects of their prolonged usage, mainly caused by CO2 rebreathing. Herein we report the development of a sensing platform for gaseous CO2 real-time determination inside FFP2 facemasks. The system consists of an opto-chemical sensor combined with a flexible, battery-less, near-field-enabled tag with resolution and limit of detection of 103 and 140 ppm respectively, and sensor lifetime of 8 h, which is comparable with recommended FFP2 facemask usage times. We include a custom smartphone application for wireless powering, data processing, alert management, results displaying and sharing. Through performance tests during daily activity and exercise monitoring, we demonstrate its utility for non-invasive, wearable health assessment and its potential applicability for preclinical research and diagnostics.

Processing parameters involved in the development of texture and tyrosine precipitates in dry-cured ham: Modelisation of texture development.

The aim of this study was to quantify the effects of different processing parameters on texture development and the incidence of white film and tyrosine crystals in dry-cured ham. Hams were dry-salted for 0.65, 0.8 or 1.0 days/kg. After drying for 45 days at 5 °C, they were dried at 10, 15 or 20 °C until reaching 33% weight loss and, thereafter, dried at 25 °C until reaching 36 or 40% weight loss. The salting time, drying temperature and target weight loss significantly affected the texture and incidence of white film and tyrosine crystals. A beneficial effect of drying at 20 °C on texture was found, which was especially important for low target weight loss (33%). Besides, hams dried at 20 °C and those with 40% weight loss showed higher incidence of tyrosine crystals. Contour plots and predictive models for texture can be used to define optimal processing parameters.

Instrumental texture analysis on the surface of dry-cured ham to define the end of the process.

The end of the elaboration process of dry-cured ham is currently decided by product weight loss and/or by an expert who carries out an evaluation of the tactile texture on the surface. The objective of this study was to define the optimal measurement conditions of an instrumental texture analysis on the surface of the dry-cured ham (ITAS), to define the end of process. 120 dry-cured hams were classified by experts into Hard (appropriate) or Soft (non-appropriate) texture groups and used to perform compression tests using different probes on three anatomical positions. Results showed that the small probe in position 2 gave the most discriminant conditions, providing representative information of the internal texture. Although classification using only weight loss was possible with an accuracy rate of 80.4% or 66.7% depending on the weight loss, the maximum classification accuracy was obtained when using ITAS in combination with weight loss. Further studies at industrial level are needed.

First Report of Arabis mosaic virus on Grapevine in Spain.

Arabis mosaic virus (ArMV; genus Nepovirus, family Comoviridae) is one of several nepoviruses responsible for infectious degeneration disease of grapevines in Europe (3). The first occurrence in Spain, in the summer of 2007, was found in Val de Salnés, Rias Baixas appellation, Galice on 25-year-old vines of the Albariño variety grafted onto an unidentified rootstock and showing leaf yellowing. The second finding was in the spring of 2008 in Barriobusto, Rioja appellation, Basque Country on 30-year-old vines of Tempranillo variety grafted onto 41B rootstock. In this case, no obvious foliar symptoms were observed but fruit set was very poor. Positive ELISA results were obtained at two different laboratories using antibodies to ArMV obtained from two companies (BIOREBA, Reinach, Switzerland and Sediag, Longvic, France). At a third lab, the presence of ArMV was further confirmed by reverse transcription (RT)-nested PCR using the primers described by Bertolini et al. (1). External primers ArMV 1 and ArMV 2 amplified a fragment of 340 bp from the coat protein region of the virus and internal primers ArMV i1 and ArMV i2 amplified a fragment of 203 bp. The specificity of the amplicons was subsequently confirmed by sequencing and comparison with other ArMV isolates available in the GenBank, EMBL, and DDBJ databases. Alignment performed using Blastn showed 85% nucleotide sequence identity with ArMV isolate NW (Accession No. AY017339). ELISA revealed co-infection with GLRaV-1 in Galice, GLRaV-3 in Rioja, and GFkV at both sites; these other viruses being common in their respective appellations. ArMV could be mechanically transmitted from rooted cuttings onto Chenopodium amaranticolor with an average of a 46% success rate (1:10 tissue/buffer ratio; [2]), but the range was very wide (0 to 100%) and dependent on the individual source vine. No statistical differences were found between nicotine or phosphate buffer for extraction or when using shoot tips or root tips as a source of virus (Fisher's exact test). Infection in C. amaranticolor was symptomless, but detectable by ELISA, and systemic. The Galician grapevine was an isolated plant, replanted on the spot of a dead one. Xiphinema diversicaudatum, the nematode vector of ArMV, was found in the vineyard soil. Only two ArMV-positive vines were found among 1,993 plants analyzed in Galice from 2005 to 2007 (no field data available for the second finding). In Rioja, one positive vine was found in a random sample of 74 vines from two different vineyards. Further testing of the neighboring vines indicated that one of the adjacent plants was also infected. This minimal spread since the vineyard was planted is suggestive of a lack of vectored transmission. In Spain as a whole, the virus seems to be rare and associated with the Atlantic biogeographic region. Both vineyards were planted before certified material became widely available. Currently, statutory testing of grapevine propagation material should prevent further spread. References: (1) E. Bertolini et al. Phytopathology 93:286, 2003. (2) G. P. Martelli, ed. Graft-Transmissible Diseases of Grapevines. Handbook for Detection and Diagnosis, FAO, Rome, 1993. (3) G. P. Martelli and E. Boudon-Padieu. Directory of Infectious Diseases of Grapevines and Viruses and Virus-like Diseases of the Grapevine. Bibliographic Report 1998-2004, CIHEAM, Paris, 2006.

Optical portable instrument for the determination of CO2 in indoor environments.

A portable device based on a colorimetric sensor to determine the atmospheric level of CO2 gas is presented in this work. The system is based on a low-cost, low-power System on a Chip (SoC) microcontroller with integrated Wi-Fi. A user-friendly application was developed to monitor and log the CO2 measurements when the system is connected to a Wi-Fi network. The sensing membrane is directly deposited on the surface of the colour detector, thus reducing the complexity of the system. This sensing membrane is formed by a pH indicator α-naphtholphthalein, tetramethylammonium hydroxide pentahydrate, 1-ethyl-3-methylimidazolium tetrafluoroborate, Tween 20 and hydroxypropyl methylcellulose as the hydrophilic polymer. The system has been fully characterized, obtaining response and recovery times of 1.3 and 2.5 s, respectively, a limit of detection of 51 ppm, and an average resolution of 6.3 ppm. This portable device was applied for the in-situ determination of CO2 gas in the atmosphere inside classrooms in several secondary schools. The measurements were taken during complete workdays and the results were statistically compared with the same measurements taken using a commercially available non-dispersive infra-red (NDIR) device. No significant statistical differences were found between the results obtained using both devices. A complete statistical treatment of the measurements made with the proposed portable device was carried out. The obtained results show that the concentration of CO2 gas in some schools was higher than the desired concentration, with regard to influencing the student's health, safety, productivity and comfort. This demonstrates the need to control this parameter to ensure appropriate indoor environmental quality (IEQ).

A novel hybridization approach for detection of citrus viroids.

Citrus plants are natural hosts of several viroid species all belonging to the family Pospiviroidae. Previous attempts to detect viroids from field-grown species and cultivars yielded erratic results unless analyses were performed using Etrog citron a secondary bio-amplification host. To overcome the use of Etrog citron a number of RT-PCR approaches have been proposed with different degrees of success. Here we report the suitability of an easy to handle northern hybridization protocol for viroid detection of samples collected from field-grown citrus species and cultivars. The protocol involves: (i) Nucleic acid preparations from bark tissue samples collected from field-grown trees regardless of the growing season and storage conditions; (ii) Separation in 5% PAGE or 1% agarose, blotting to membrane and fixing; (iii) Hybridization with viroid-specific DIG-labelled probes and detection with anti-DIG-alkaline phosphatase conjugate and autoradiography with the CSPD substrate. The method has been tested with viroid-infected trees of sweet orange, lemon, mandarin, grapefruit, sour orange, Swingle citrumello, Tahiti lime and Mexican lime. This novel hybridization approach is extremely sensitive, easy to handle and shortens the time needed for reliable viroid indexing tests. The suitability of PCR generated DIG-labelled probes and the sensitivity achieved when the samples are separated and blotted from non-denaturing gels are discussed.

Direct interaction of colloidal nanostructured ZnO and SnO2 with NO and SO2.

A novel and easy synthesis pathway of small SnO2 nanoparticles is reported. The method consists of the spontaneous hydrolysis of SnCl4 x 5H2O in dimethyl sulfoxide (DMSO), containing 3% water, at room temperature. The structure of the SnO2 nanocrystals corresponds to that of the cassiterite phase, as shown by powder X-ray diffraction and HR-TEM. The UV-visible electronic absorption and emission spectra of the SnO2 colloids are discussed. The reactions of NO(g) and SO2(g) with ZnO (wurtzite phase) and SnO2 nanocolloids are studied. The interaction of NO with ZnO nanoparticles generates the dissolution of the particles and it is quite probable that NO3(-1), NOs(-2), N2O and N2 are formed; while its contact with SO2 probably yields SO4(-2), SO3(-2) and also the dissolution of the particles is observed. When these gases are reacted with SnO2, then NO3(-1), NO2(-1), SO3(-2) and SO4(-2), were respectively obtained.

Effects of plant alkylphenols on cytokine production, tyrosine nitration and inflammatory damage in the efferent phase of contact hypersensitivity.

BACKGROUND AND PURPOSE: The phenolic compounds isoprenylhydroquinone glucoside (IHG), 3,5-dicaffeoylquinic acid (DCA), and its methyl ester (DCE) have previously been shown to inhibit both contact hypersensitivity (CHS) and peroxynitrite reactivity. The present work seeks to establish a relationship between the anti-inflammatory effect and the release of cytokines and tyrosine nitration in skin. EXPERIMENTAL APPROACH: Murine CHS was developed by means of sensitization and challenge with dinitrofluorobenzene (DNFB) or oxazolone. Ear swelling was measured 24 and 96 h after challenge. Interleukin (IL)-1beta, IL-4, and tumour necrosis factor (TNF)-alpha were measured by ELISA; and the expression of inducible nitric oxide synthase (iNOS) was detected by Western blotting. Histological samples were analysed for 3-nitrotyrosine. KEY RESULTS: In the oxazolone model, DCE reduced the 24 h swelling by 54% whereas the effect of DCA was lower (40% inhibition). All the test compounds reduced IL-1beta values 24 h after challenge with DNFB or oxazolone, DCE particularly inhibited IL-4 production (74% and 78%, respectively; P<0.01). Tyrosine nitration was also markedly reduced by DCE. In general, the test compounds limited the presence of polymorphonuclear (PMN) leukocytes in the skin. CONCLUSIONS AND IMPLICATIONS: These results suggest that the effect of 3,5-dicaffeoylquinic esters on CHS is associated with a decrease in the production of interleukins, but not with the inhibition of iNOS expression. Moreover, esterification of the carboxyl group at C-1 enhanced protection against tyrosine nitration in the skin.

[Painful ophthalmoplegia (pseudotumor of the orbit and Tolosa-Hunt syndrome)]. / Oftalmoplejía dolorosa (pseudotumor y síndrome de Tolosa-Hunt).

INTRODUCTION: The Tolosa-Hunt Syndrome (THS) and the Pseudotumor of the Orbit (PTO) each have three common clinical hallmarks: unilateral periorbital pain, cranial nerve palsies and a fast response to corticosteroid therapy. CASE REPORT: 48-year-old female with a right VI nerve paralysis, later develops a right III nerve paralysis. It is important to be able to differentiate between THS and PTO, and this is done on the basis of the complementary tests. DISCUSSION: Apart from neuroradiologic findings, the clinical presentation and histopathology of these two conditions are very similar, making it difficult to distinguish between them. Aspects common to both pathological processes are discussed.

Soymilk fermentation by Enterococcus faecalis VB43 leads to reduction in the immunoreactivity of allergenic proteins ß-conglycinin (7S) and glycinin (11S).

Food allergies represent a serious problem affecting human health and soy proteins rank among the most allergenic proteins from food origin. The proteolytic enzymes produced by lactic acid bacteria (LAB) can hydrolyse the major allergens present in soybean, reducing their immunoreactivity. Many studies have reported the ability of LAB to ferment soy-based products; while the majority of them focus on the improvement of the sensory characteristics and functionality of soy proteins, a lack of information about the role of lactic fermentation in the reduction of immunoreactivity of these proteins exists. The aim of the present study was to evaluate the capability of the proteolytic strain Enterococcus faecalis VB43 to hydrolyse the main allergenic proteins present in soymilk and to determine the immunoreactivity of the obtained hydrolysates. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) results of fermented soymilk demonstrated complete hydrolysis of the ß-subunit from ß-conglycinin and the acidic polypeptide from glycinin. Reversed phase high performance liquid chromatography (RP-HPLC) analysis of the peptides released after hydrolysis revealed the appearance of new peptides and the disappearance of non-hydrolysed proteins, indicating extensive hydrolysis of the substrate. Results from competitive enzyme-linked immunosorbent assay (ELISA) tests clearly indicated a reduction in the immunoreactivity (more than one logarithmic unit) in the fermented sample as compared to the non-fermented control. Our results suggest that the soymilk fermented by E. faecalis VB43 may induce lower allergic responses in sensitive individuals. The strain E. faecalis VB43 may be considered as an excellent candidate to efficiently reduce the immunoreactivity of soymilk proteins.

Moles quantification in liquid samples by Raman spectroscopy.

The mole is a unit of measurement that expresses amounts of a chemical substance. Its importance lies in that the mass and the number of molecules of a substance can be determined with this value. In this work, we suggest a mathematical expression that relates the number of moles of the sample studied with the Raman signal and the experimental parameters used. In other words, with this mathematical expression it is possible to obtain quantitative information in a simple manner from Raman spectra. We have applied this method to different samples and we have observed an excellent correlation between the experimental and expected data.