ΣΥΓΚΡΙΣΗ ΚΑΙ ΕΠΙΚΥΡΩΣΗ ΠΟΛΥΫΠΟΛΕΙΜΜΑΤΙΚΩΝ ΜΕΘΟΔΩΝ ΠΡΟΣΔΙΟΡΙΣΜΟΥ ΚΑΤΑΛΟΙΠΩΝ ΓΕΩΡΓΙΚΩΝ ΦΑΡΜΑΚΩΝ ΣΕ ΧΥΜΟΥΣ ΜΕ ΤΕΧΝΙΚΕΣ ΥΓΡΟΧΡΩΜΑΤΟΓΡΑΦΙΑΣ-ΦΑΣΜΑΤΟΜΕΤΡΙΑΣ ΜΑΖΩΝ ΣΕ ΣΕΙΡΑ (LC-MS/MS) ΚΑΙ ΑΕΡΙΟΧΡΩΜΑΤΟΓΡΑΦΙΑΣ-ΦΑΣΜΑΤΟΜΕΤΡΙΑΣ ΜΑΖΩΝ ΠΑΓΙΔΑΣ ΙΟΝΤΩΝ (GC-MSn)

Στην παρούσα εργασία συγκρίθηκαν και επικυρώθηκαν τρεις πολυ¬ϋπο¬λειμ¬ματικές μέθοδοι για τον προσδιορισμό καταλοίπων φυτο¬φα¬ρμάκων σε δεί¬γματα χυμών. Συγκεκριμένα, έγινε μελέτη μεθόδων α) με άμεση έγχυση του δείγματος (δηλαδή χωρίς να προηγείται προκατεργασία του δείγματος χυμού, παρά μόνο αραίωση και διήθηση) β) με προκατεργασία χρη¬σιμοποιώντας εκχύλιση στερεής φάσης (SPE) με υλικό Oasis HLB και, γ) με προκατεργασία χρη¬σιμοποιώντας τη μέθοδο QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe). Τα εκχυλίσματα που προκύπτουν από την προκατεργασία εγχύθηκαν σε συστήματα υγροχρωματογραφίας-φασματομετρίας μαζών σε σειρά (LC-MS/MS) και αεριοχρωματογραφίας-φασματομετρίας μαζών παγίδας ιόντων (GC-MSn) για τον προσδιορισμό 203 και 85 ενώσεων, αντίστοιχα. Στη συνέχεια, οι τρεις μέθοδοι επικυρώθηκαν και αξιολογήθηκαν, σύμφωνα με την οδηγία της Ευρωπαϊκής Επιτροπής SANTE/11813/2017, ως προς τα χαρακτηριστικά της γραμ¬μικότητας, ορθότητας και πιστότητας (τόσο υπό συνθή¬κες επα¬να¬λη¬ψιμότητας όσο και ενδοεργαστηριακής αναπαρα¬γωγι¬μότητας), προσδιο¬ρίσθηκαν τα όρια ανίχνευσης και ποσοτικοποίησης και εκτι¬μήθηκε το φαι¬νόμε¬νο μήτρας. Η ταυτοποίηση και η ποσοτικοποίηση των αναλυτών-στόχων πραγματοποιήθηκε με LC-ESI-MS/MS με ιοντισμό με ηλεκτροδιάχυση (LC-ESI-MS/MS) και με αεριοχρωματογραφία-φασματομετρία μαζών με ιονισμό με πρόσκρουση ηλεκτρονίων θετικής και αρνητικής πολικότητας (GC-EI-MSn) με λειτουργία σάρωσης δύο επιλεγμένων αντίδρασεων (ΜRM) ανά ουσία. Εξαγόμενο συμπέρασμα της παραπάνω μελέτης αποτελεί η εκτίμηση ότι η μέθοδος άμεσης έγχυσης ενώ θα μπορούσε να είναι μια χρήσιμη εναλλακτική προσέγγιση για την πολυϋπολειμματική ανάλυση χυμών δεν παρουσιάζει την απαιτούμενη απόδοση σε μεγάλο εύρος ουσιών. Η καταλληλότερη μέθοδος από αυτές που εξετάστηκαν είναι η μέθοδος QuEChERS,σημείωσε την καλύτερη απόδοση κατά την επικύρωση, είναι απλή στην εφαρμογή της και δεν απαιτεί μεγάλη δαπάνη χρόνου και κόστους από ένα εργαστήριο. Τέλος η μέθοδος SPE αποτελεί μία καλή επιλογή στην περίπτωση που απαιτείται προσυγκέντρωση.In this work three multi-residue methods for the determination of pesticide residues in juice samples were developed, validated and compared. The sample preparation was performed using three different approaches; i) the Quick, Easy, Cheap, Effective, Rugged, Safe (QuEChERS) method with citrate, ii) solid phase extraction (SPE) using OASIS HLB cartridges, and, iii) the dilute-and-shoot (direct injection) method. Extracts obtained using the different extraction techniques were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) and gas chromatography-ion trap mass spectrometry (GC-MSn) for the detection and quantification 203 compounds and 85 compounds, respectively. The three methods were validated according to the European Union guidelines for pesticide residues SANTE/11813/2017 for linearity, accuracy and precision (repeatability and reproducibility) while the limit of detection and quantification were evaluated along with the matrix effect. The detection and the quantification of the analytes was performed with LC-MS/MS with electro-spray ionisation (LC-ESI-MS/MS) and GC-MSn with electron ionisation (GC-EI-MSn) in the multiple reaction monitoring (MRM) mode. The outcome of the aforementioned study is that the direct injection method would have a potential as a useful alternative approach, but it is not performing well for a variety of substances. The most suitable method for the determination of juices is QuEChERS as it was performed best during validation; it is a simple, cost effective and time efficient implementation. Finally, the SPE method can be an option in the case that preconcentration is essential

Heat and Cold-Stressed Individuals of <i>Pistacia lentiscus</i> (Mastic Tree) Do Modify Their Secreting Profile

Seedlings from the germinated seeds of Pistacia lentiscus were cultured in plant growth chambers for three months. Then, the plants were separated into three groups. Each group was cultured under different conditions. The first group was left to grow under normal Mediterranean conditions, as those recorded in spring. The other group was subjected to a ten-day heat stress while the last one also suffered a cold stress for ten days. The anatomical features of the leaves (leaf thickness, epidermal cell thickness, number of palisade layers, and development) between these three groups differed. The stressed plants accumulated large amounts of phenolics within their mesophyll cells. The biomass of the cold-stressed plants was minor, while it was high for the control plants. The oxidative stress was hardly detectable in the leaves of the control plants, while their heat-stressed counterparts suffered the highest concentration of reactive oxygen species. Differences concerning the absorption spectra of the three groups of leaves were not significant. An interesting incompatibility between the three groups concerned the expression of L-Dopa Decarboxylase, which climbed significantly in the heat-stressed plants. Finally, an interesting variation was observed concerning the concentrations of some biogenic amines/amino acids. This variation can be correlated to the other stress-induced reactions of the plants and, in some cases, was impressive. In conclusion, environmental stress can shift Pistacia lentiscus’ metabolism to synthesize different biogenic products, which can be considered as exploitable for the pharmaceutical or food industry

Postprandial Glucose and Gastrointestinal Hormone Responses of Healthy Subjects to Wheat Biscuits Enriched with L-Arginine or Branched-Chain Amino Acids of Plant Origin

The study investigates the effects of wheat biscuits supplemented with plant flours originating from legumes/seeds enriched either in L-arginine (L-arg) or branched-chain amino acids (BCAAs) on postprandial glucose response of healthy subjects. Gastrointestinal hormone and amino acid responses as well as subjective appetite sensations are also evaluated. Subjects consumed wheat-based biscuits, enriched either in L-arg (ArgB) or BCAAs (BCAAsB) or a conventional wheat biscuit (CB) or a glucose solution (GS) in an acute randomized crossover design. Responses of glucose, insulin, ghrelin, glucagon-like peptide-1 (GLP-1), peptide YY (PYY) and glicentin, as well as those of L-arginine, L-leucine, L-isoleucine and L-valine, were evaluated over 180 min. Consumption of ArgB and BCAAsB elicited lower glucose iAUC compared to GS (p p p < 0.05 compared to CB). BCAAsB and ArgB increased postprandial amino acid concentrations and caused stronger satiety effects compared to CB. Increasing protein content of wheat biscuits with supplementation of plant flours originating from legumes/seeds decreases postprandial glycemia and provides with healthier snack alternatives which can easily be incorporated into diet

Prospective observational cohort study on grading the severity of postoperative complications in global surgery research

Background The Clavien–Dindo classification is perhaps the most widely used approach for reporting postoperative complications in clinical trials. This system classifies complication severity by the treatment provided. However, it is unclear whether the Clavien–Dindo system can be used internationally in studies across differing healthcare systems in high- (HICs) and low- and middle-income countries (LMICs). Methods This was a secondary analysis of the International Surgical Outcomes Study (ISOS), a prospective observational cohort study of elective surgery in adults. Data collection occurred over a 7-day period. Severity of complications was graded using Clavien–Dindo and the simpler ISOS grading (mild, moderate or severe, based on guided investigator judgement). Severity grading was compared using the intraclass correlation coefficient (ICC). Data are presented as frequencies and ICC values (with 95 per cent c.i.). The analysis was stratified by income status of the country, comparing HICs with LMICs. Results A total of 44 814 patients were recruited from 474 hospitals in 27 countries (19 HICs and 8 LMICs). Some 7508 patients (16·8 per cent) experienced at least one postoperative complication, equivalent to 11 664 complications in total. Using the ISOS classification, 5504 of 11 664 complications (47·2 per cent) were graded as mild, 4244 (36·4 per cent) as moderate and 1916 (16·4 per cent) as severe. Using Clavien–Dindo, 6781 of 11 664 complications (58·1 per cent) were graded as I or II, 1740 (14·9 per cent) as III, 2408 (20·6 per cent) as IV and 735 (6·3 per cent) as V. Agreement between classification systems was poor overall (ICC 0·41, 95 per cent c.i. 0·20 to 0·55), and in LMICs (ICC 0·23, 0·05 to 0·38) and HICs (ICC 0·46, 0·25 to 0·59). Conclusion Caution is recommended when using a treatment approach to grade complications in global surgery studies, as this may introduce bias unintentionally

Critical care admission following elective surgery was not associated with survival benefit: prospective analysis of data from 27 countries

This was an investigator initiated study funded by Nestle Health Sciences through an unrestricted research grant, and by a National Institute for Health Research (UK) Professorship held by RP. The study was sponsored by Queen Mary University of London

The surgical safety checklist and patient outcomes after surgery: a prospective observational cohort study, systematic review and meta-analysis

© 2017 British Journal of Anaesthesia Background: The surgical safety checklist is widely used to improve the quality of perioperative care. However, clinicians continue to debate the clinical effectiveness of this tool. Methods: Prospective analysis of data from the International Surgical Outcomes Study (ISOS), an international observational study of elective in-patient surgery, accompanied by a systematic review and meta-analysis of published literature. The exposure was surgical safety checklist use. The primary outcome was in-hospital mortality and the secondary outcome was postoperative complications. In the ISOS cohort, a multivariable multi-level generalized linear model was used to test associations. To further contextualise these findings, we included the results from the ISOS cohort in a meta-analysis. Results are reported as odds ratios (OR) with 95% confidence intervals. Results: We included 44 814 patients from 497 hospitals in 27 countries in the ISOS analysis. There were 40 245 (89.8%) patients exposed to the checklist, whilst 7508 (16.8%) sustained ≥1 postoperative complications and 207 (0.5%) died before hospital discharge. Checklist exposure was associated with reduced mortality [odds ratio (OR) 0.49 (0.32–0.77); P\u3c0.01], but no difference in complication rates [OR 1.02 (0.88–1.19); P=0.75]. In a systematic review, we screened 3732 records and identified 11 eligible studies of 453 292 patients including the ISOS cohort. Checklist exposure was associated with both reduced postoperative mortality [OR 0.75 (0.62–0.92); P\u3c0.01; I2=87%] and reduced complication rates [OR 0.73 (0.61–0.88); P\u3c0.01; I2=89%). Conclusions: Patients exposed to a surgical safety checklist experience better postoperative outcomes, but this could simply reflect wider quality of care in hospitals where checklist use is routine