Determination of fungicide pyrimethanil in grapes, must, fermenting must and wine.

BACKGROUND: This study determined the evolution of pyrimethanil, a fungicide commonly used to control Botrytis cinerea, throughout the winemaking process in grapes, must, fermenting must and wine. Tempranillo grapevines were treated with pyrimethanil according to both good Agricultural Practices (GAP) and Critical Agricultural Practices (CAP). Fermentation was carried out in an experimental winery. Grape analysis was based on an ethyl acetate extraction method. Samples from fermentation were analysed by solid phase extraction. The determination was carried out by gas chromatography with nitrogen-phosphorus detection (GC-NPD) and additionally confirmed by gas chromatography/mass spectrometry (GC/MS). RESULTS: Pyrimethanil residues were at least ten times greater in grapes treated 7 days before harvest than in those treated respecting the safety period (21 days). The amount of pyrimethanil in grapes treated under GAP was below the maximum residue limit (5 mg kg(-1)). The level of pyrimethanil decreased during fermentation in both treatments. In the fermentation of grapes treated according to CAP, the pyrimethanil concentration was reduced by over 50% on the first day and then remained constant until the end of the fermentation process. For grapes treated in compliance with GAP, the amount of pyrimethanil decreased to a level below the limit of detection in the bottled wine. CONCLUSION: The described methods for grapes, must, fermenting must and wine gave good recoveries, linearity, precision and accuracy. They were also highly sensitive in avoiding matrix effects. Pyrimethanil residues found in treated grapes were higher in skin than in pulp. The amount of pyrimethanil decreased during fermentation by degradation and/or adsorption. For grapes treated according to GAP, residues disappeared in the final bottled wine. The decrease observed in the final bottled wine may be caused by diverse oenological practices and technologies such us malolactic fermentation, racking and settling.

Optimization and validation of a simple and fast method for the determination of fungicides in must and wine samples by SPE and GC/MS.

A rapid, simple, and low-cost method based on SPE was optimized and validated for simultaneous determination of eight fungicides belonging to different chemical classes in must and wine. The method involves extraction of 10 mL of must or wine samples with a C18 cartridge using 5 mL of dichloromethane as the elution solvent. Separation and final determination of the fungicides (vinclozolin, dichlofluanid, penconazol, captan, quinoxyfen, fluquinconazol, boscalid, and pyraclostrobin) was performed by GC coupled to single quadrupole MS. Recoveries at 10, 50, and 100 microg/L were between 71 and 106% in both matrixes for the fungicides evaluated. The calculated LOQ ranged from 1.5 to 3.4 microg/L in must and 1.1 to 3.8 microg/L in wine. Matrix effects observed for wine and must samples were overcome by using matrix-matched calibration. The developed method was linear at concentrations within the tested interval, with coefficients of determination higher than 0.999. The expanded uncertainties at 10 microg/L were <20% for all analytes. Intralaboratory precision in terms of the Horwitz ratio of the fungicides evaluated was below 0.5, suggesting the ruggedness of the method. The proposed method was applied to determine fungicide residues in must samples obtained from red grapes treated with two new commercial formulations, as well as in their corresponding final wines.

Anemia and Outcome in Outpatients With Peripheral Artery Disease.

The influence of anemia on outcome in stable outpatients with peripheral artery disease (PAD) has not been consistently investigated. We used data from the Factores de Riesgo y ENfermedad Arterial (FRENA) Registry to compare ischemic events and mortality rates in stable outpatients with symptomatic PAD and anemia. Of 1663 patients with PAD, 208 (12.5%) had anemia. Over 18 months, patients with anemia had a higher rate of myocardial infarction (MI; rate ratio [RR]: 2.10; 95% confidence interval [CI]: 1.04-3.99), limb amputation (RR: 2.98; 95%CI: 1.70-5.05), and higher mortality (RR: 3.58; 95%CI: 2.39-5.28) than those without anemia. The rates of ischemic stroke (RR: 0.75; 95%CI: 0.23-1.93) and major bleeding (RR: 0.93; 95%CI: 0.15-3.51) were similar. On multivariable analysis, anemia was associated with an increased risk to die (hazard ratio [HR]: 2.32; 95%CI: 1.53-3.50) but not to develop MI (HR: 1.49; 95%CI: 0.73-3.05) or to have limb amputation (HR: 1.49; 95%CI: 0.86-2.59). In stable outpatients with PAD, anemia was associated with increased mortality but not with an increased rate of subsequent ischemic events or major bleeding.

Dissipation of Fungicide Residues during Winemaking and Their Effects on Fermentation and the Volatile Composition of Wines.

The effects of four fungicides commonly used for the control of fungal diseases in vines and grapes in the course of winemaking were tested. The concentration of fungicide residues was monitored throughout the process to establish their kinetics of dissipation. In all cases the percentages of dissipation were >68%, which shows the detoxificant effect of the winemaking process. On the other hand, the effect of the fungicide residues on the aroma composition of Tempranillo red wines was tested. To evaluate possible modifications on the aroma profile of wines, seven odorant series (ripe fruits, fresh fruits, lactic, floral, spicy, vinous, and herbaceous) were built from the odor activity values (OAVs) obtained for each volatile compound. Ripe fruits and fresh fruits were the major aromatic attributes in all Tempranillo red wines. These two odorant series registered the highest variations in their total OAVs with respect to the control wine, especially with the application of boscalid + kresoxim-methyl into vines, leading to a decrease in the ripe fruit and fresh fruit nuances of the resulting wines. Moreover, when the effect of these fungicides on the aroma of Tempranillo red wines was compared throughout two years (2012 and 2013), wines elaborated from grapes treated in the field with boscalid + kresoxim-methyl in 2013 displayed the highest variation in aroma profile with respect to control wine.

Effect of cluster thinning and prohexadione calcium applications on phenolic composition and sensory properties of red wines.

The overall objective of this study was to investigate the effect of manual cluster thinning (CT) and the application of the growth regulator Prohexadione calcium (ProCa) on the phenolic composition and the sensory profile of Tempranillo and Grenache wines produced from treated vines in La Rioja (Spain). ProCa was applied at preblooming and CT was carried out at veraison in two consecutive years. Different physicochemical parameters and analyses of phenolic compounds were carried out in control, CT and ProCa grapes and wines and wine sensory was performed. Thinning treatments decreased crop yield, besides ProCa application reduced berry size, and berry weight. Color and phenolic composition of Grenache and Tempranillo wines in general were affected by thinning treatments, with an increase in anthocyanin, flavanol and flavonol concentrations. In sensory analysis, wines obtained from thinned vines presented higher values for several aromatic (e.g., white and yellow fruits, fresh flowers) and taste attributes (i.e., astringency, bitternes, persistence). CT and ProCa treatments resulted in an improvement in wine quality. In general, similar results in phenolic composition, sensory properties and quality of wines were obtained by manual and chemical cluster thinning. ProCa as a growth regulator may be an option for a quality vitiviniculture.

Endoscopic ultrasonography in patients with large gastric folds at endoscopy and biopsies negative for malignancy: predictors of malignant disease and clinical impact.

OBJECTIVES: The aims of the study were to assess in patients with large gastric folds at endoscopy and endoscopic biopsies that tested negative for malignancy: (i) the predictive variables of malignancy in endoscopic ultrasonography (EUS), (ii) the impact of EUS. METHODS: Patients with this condition evaluated with radial EUS (following the standard study protocol of our Unit in these patients) during a 5-yr period were included. Nine variables were evaluated as possible predictors of malignancy (chi2, t-test, and logistic regression procedure) using pathology results, or a minimum of 2 yr of follow-up as gold standard. Likelihood ratios (LR), pre-EUS, and post-EUS probabilities for the diagnosis of malignancy were used to assess the clinical impact of the technique. RESULTS: Sixty-one patients were included (40 benign and 21 malignant). Predictive factors of malignancy were as follows: thickened gastric wall, thickened deep layers, impaired gastric distension, loss of the wall structure, and presence of ascites or lymph nodes, whereas the enlargement of superficial layers was a predictor of a benign condition. The enlargement of deep layers, as assessed by EUS was the only independent predictive factor for malignancy. When using this parameter, the pre-EUS probability of malignancy in our series (34%) increased up to 95% when EUS suggested malignancy (positive LR = 45), whereas it decreased to 4.7% when EUS precluded this diagnosis (negative LR = 0.102). CONCLUSIONS: (i) The enlargement of deep layers is the only independent predictive factor for malignancy in patients with large gastric folds at endoscopy and biopsies testing negative for malignancy and (ii) EUS has a high clinical impact in these patients.

Usefulness of EUS-guided fine needle aspiration (EUS-FNA) in the diagnosis of functioning neuroendocrine tumors.

BACKGROUND: Localization of neuroendocrine tumors may be challenging. The role of EUS-FNA in this setting is unknown. METHODS: Ten patients with clinically suspected functioning neuroendocrine tumors (hormonal disturbances) underwent EUS-FNA to determine the location and to confirm the diagnosis cytologically. OBSERVATIONS: EUS identified 14 tumors in these 10 patients. In all but one patient CT did not demonstrate the tumor or missed at least one of multiple lesions. Mean tumor size was 12 mm (range 4-25 mm). Tumor locations were pancreas (n = 13) and duodenal wall (n = 1). Eleven of the 14 detected lesions were aspirated under EUS with accurate diagnosis in all cases. Surgical confirmation of EUS-FNA findings was available in 7 patients. There was no complication of EUS-FNA. CONCLUSIONS: EUS is a highly accurate technique for visualization of small functioning neuroendocrine tumors not evident on CT and for identification of patients with multiple lesions. EUS-FNA safely provides cytologic confirmation with high accuracy in these patients.

Ultrasonografía endoscópica: aplicabilidad diagnóstica y terapéutica / Clinical use and indications of endoscopic ultrasonography

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