Wine and grape marc spirits metabolomics.

INTRODUCTION: Mass spectrometry (MS)-based and nuclear magnetic resonance (NMR) spectroscopic analyses play a key role in the field of metabolomics due to their important advantages. The use of metabolomics in wine and grape marc spirits allows a more holistic perspective in monitoring and gaining information on the making processes and thus it can assist on the improvement of their quality. OBJECTIVES: This review surveys the latest metabolomics approaches for wine and grape marc spirits with a focus on the description of MS-based and NMR spectroscopic analytical techniques. METHODS: We reviewed the literature to identify metabolomic studies of wine and grape marc spirits that were published until the end of 2017, with the key term combinations of 'metabolomics', 'wine' and 'grape marc spirits'. Through the reference lists from these studies, additional articles were identified. RESULTS: The results of this review showed that the application of different metabolomics approaches has significantly increased the knowledge of wine metabolome and grape marc spirits; however there is not yet a single analytical platform that can completely separate, detect and identify all metabolites in one analysis. CONCLUSIONS: The authentication and quality control of wines and grape marc spirits has to be taken with caution, since the product's chemical composition could be affected by many factors. Despite intrinsic limitations, NMR spectroscopy and MS based strategies remain the key analytical methods in metabolomics studies. Authenticity, traceability and health issues related to their consumption are the major research initiatives in wine and grape marc spirits metabolomics analysis.

Early KPC-Producing Klebsiella pneumoniae Bacteremia among Intensive Care Unit Patients Non-Colonized upon Admission.

Among 140 patients colonized by KPC-producing Klebsiella pneumoniae (KPC-Kp) between fourth and seventh day of Intensive Care Unit stay, 24 developed bacteraemia immediately after colonization. Colistin-resistance of the colonizing isolate was the factor significantly associated with early KPC-Kp bacteraemia (P < 0.001; OR 6.6, 95% CI 2.4-18.4), a worrisome finding since infections by colistin-resistant isolates is associated with increased mortality due to limited remaining therapeutic options.

Myasthenia Gravis and Abdominal Aortic Aneurysm: A Rare Combination.

Abdominal aortic aneurysm in a patient with myasthenia gravis (MG) is extremely rare. We present a 64-year-old male with MG and an asymptomatic abdominal aortic aneurysm treated endovascularly. After extubation, he suffered a cardiac arrest due to an acute myocardial infarction. Cardiopulmonary resuscitation and a primary coronary angioplasty led to a satisfactory outcome. Special care is needed due to higher rates of postoperative complications in these patients.

Study of the naphthalene-2,3-dicarboxaldehyde pre-column derivatization of biogenic mono- and diamines in mixture and fluorescence-HPLC determination.

A new fluorescence-HPLC method was developed for the simultaneous determination of eight biogenic monoamines (histamine, methylamine, tyramine, ethylamine, propylamine, tryptamine, 2-phenylethylamine, isoamylamine) and two biogenic diamines (putrescine, cadaverine) in the presence of heptylamine as the internal standard. The amines were pre-column derivatized with naphthalene-2,3-dicarboxaldehyde in the presence of cyanide ion as the nucleophile. The effect of the derivatization reaction conditions on the reaction yield was investigated. The derivatives were separated on an Inertsil ODS-3 column (250 × 4mm i.d., 5 µm) using gradient elution and detected fluorimetrically at excitation and emission wavelengths of 424 and 494 nm, respectively. Limits of detection between 0.002 and 0.4 ng, injected on-column (10-µL loop), were achieved. The within- and between-day relative standard deviations ranged between 0.2-3.4% and 0.3-4.8%, respectively. The utility of the method in assaying biogenic mono- and diamine mixtures in Greek cheeses is demonstrated. Ultrasound-assisted liquid-liquid extraction was applied prior to derivatization.

Development and validation of a liquid chromatographic method with diode array detection for the determination of anthraquinones, flavonoids and other natural dyes in aged silk.

A HPLC method coupled with diode array detector was developed and validated for the quantitation of alizarin, apigenin, carminic acid, curcumin, ellagic acid, emodin, fisetin, kaempferide, kaempferol, kermesic acid, morin, purpurin, quercetin and sulfuretin which are components of several natural dyes. 1- Hydroxyanthraquinone was selected as internal standard. The compounds were separated under gradient elution on a RP-column (Altima C18, 250 mm x 3.0 mm i.d., 5 µm) with a mobile phase consisting of solvent A: H2O + 0.1% (v/v) trifluoroacetic acid and solvent B: acetonitrile + 0.1% (v/v) trifluoroacetic acid. The method was validated in terms of linearity, limits of detection and quantitation, accuracy, precision, ruggedness and robustness and applied to the analysis of silk dyed with buckthorn (Rhamnus trees), cochineal (Dactylopius coccus Costa), madder (Rubia tinctorum L.), turmeric (Curcuma longa L.) and young fustic (Cotinus coggygria Scop). Furthermore, dyed silk samples were subjected to artificially accelerated ageing conditions induced by UV radiation. The effect of the latter on the quantities of the aforementioned compounds was monitored, except for apigenin, kermesic acid and morin.

Single run analysis of glutathione and its disulfide in food samples by liquid chromatography coupled to on-line post-column derivatization.

Glutathione and its disulfide were determined in a single run using liquid chromatography with on-line post-column derivatization and fluorimetric detection (340 nm/425 nm). The analytes were separated using a reversed-phase column capable of operating at 100% aqueous mobile phase and detected following direct on-line reaction with o-phthalaldehyde (7.5 mmol L-1) in highly basic medium (0.37 mol L-1 NaOH). The instrumental and chemical variables were carefully investigated towards high sensitivity and throughput, while special attention was paid to validating potential matrix effects. Glutathione and its disulfide could be selectively determined with respective LODs of 0.10 and 0.30 µmol L-1 in the absence of matrix effect (<6%). The endogenous content of the analytes was accurately determined in various food samples with recoveries ranging between 80 and 120% in all cases. The proposed method is reliable and promising as a generic analytical tool for the convenient estimation of the redox status of glutathione in various food matrices.

Predictors of mortality of trauma patients admitted to the ICU: a retrospective observational study☆.

BACKGROUND AND OBJECTIVES: Worldwide, trauma is one of the leading causes of morbidity and mortality. The aim of the present study is to identify the predictors of mortality of trauma patients requiring Intensive Care Unit (ICU) admission. METHODS: This retrospective study was conducted in the ICU of our institution in Greece during a six-year period (2010-215). RESULTS: Among 326 patients, trauma was caused by road traffic accidents in .5%, followed by falls (21.1%) and violence (7.4%). Thirty-day mortality was 27.3%. Multivariate analysis showed that higher New Injury Severity Score (NISS), severe head/neck injury, acute kidney injury, septic shock and hemorrhagic shock were significantly associated with mortality while higher Revised Injury Severity Classification, version II (RISC II) and the administration of enteral nutrition were associated with survival. NISS showed the higher accuracy in predicting 30-day mortality followed by RISC II, while scores based only in physiological variables had lower predictive ability. CONCLUSIONS: Increased mortality was strongly associated with the severity of the injury upon admission. Traumatic brain injury, septic shock and acute kidney injury have also been found among the strongest predictors of mortality. NISS can be considered as a statistically superior score in predicting mortality of severely injured patients.

Frequency and Significance of Maneuvers to Dissect the Distal Internal Carotid Artery During Carotid Endarterectomy.

OBJECTIVES: To describe the frequency, factors associated with, and significance of surgical dissection maneuvers of the distal internal carotid artery (ICA) during carotid endarterectomy (CEA). METHODS: In this retrospective analysis of prospectively collected information in patients undergoing CEA, we recorded information on demographics, risk factors and comorbidities, dissection maneuvers of the distal ICA, other operative variables and neurological outcome measures. RESULTS: During the period July 2008 and February 2020 inclusive, 218 consecutive patients (180 males, median age 69.5 years) underwent 240 CEAs. In 117 (48.8%) of them, CEA was performed for a symptomatic stenosis. Dissection maneuvers of the distal ICA were required in 77 cases (32.1%), including division and ligation of the sternocleidomastoid vessels in 66 cases (27.5%), mobilization of the XII cranial nerve in 69 cases (28.7%, with concomitant transection of the superior root of the ansa cervicalis in 11 cases, 4.6%) and division of the posterior belly of the digastric muscle in 8 cases (3.3%). Styloid osteotomy was not required in any case. Smoking was the single predictive factor associated with the use of an adjunctive dissection maneuver (odds ratio 2.23, p = 0.009). The use of a patch was more common in smokers (16% vs 7.1% in non-smokers, odds ratio 2.48, p = 0.05). Perioperative stroke and/or death rate was 0%, not allowing testing for associations with maneuver performance. Two patients (0.8%) developed a transient ischemic attack and 4 patients (1.7%) a cranial nerve injury (CNI), including 2 patients with recurrent laryngeal nerve palsy, diagnosed on routine laryngoscopy during planning of a contralateral CEA. There was no association between CNI and dissection of the distal ICA using an operative adjunct (p = 0.60). CONCLUSIONS: Several surgical maneuvers are often required to accomplish dissection of the distal ICA beyond the point of atherosclerotic disease. When dictated by operative findings, such maneuvers are deemed safe.

LC of sulfonamide residues in poultry muscle and eggs extracts using fluorescence pre-column derivatization and monolithic silica column.

A new, rapid, sensitive and selective HPLC method with fluorescence detection is described for the simultaneous determination of 12 sulfonamides, in the presence of putrescine as internal standard, after pre-column derivatization with fluorescamine. The drugs were separated on a Chromolith Performance RP-18 column (100x4.6 mm), using a gradient elution with a binary mobile phase of methanol/0.05 M acetate buffer (pH 3.4). Linearity of derivatization was obtained for concentrations from 3.0 to 300 microg/L in standard solutions. The whole procedure was evaluated and fully validated, according to the European Union Decision 2002/657/EC, for the determination of sulfonamides in turkey muscle and hen eggs following SPE. The LODs varied from 2 to 17 microg/kg in turkey and 2 to 15 microg/kg in egg samples. The average recoveries ranged between 96.9-108.6% in turkey muscle and 96.0-108.4% in egg samples, respectively.

A targeted approach for studying the effect of sugar bee feeding on the metabolic profile of Royal Jelly.

Royal Jelly (RJ) constitutes one of the most popular beehive products and for this reason the use of inexpensive sweeteners during its production remains an important quality issue. In the present study we report results of metabolic profiling of RJ samples obtained after the application of artificial bee-feeding using different feeding protocols. The hydrophilic content of RJ samples was assessed by applying (HILIC)UPLC-MS/MS. In total 96 crude RJ samples were analysed with the developed method. Multivariate statistical analysis revealed clear differentiation between the RJ samples obtained from control (non-fed) bees and samples obtained after feeding. In total 27 out of 57 detected molecules were statistically found to be significantly altered in the different comparisons. Among them some amino acids (e.g. tryptophan, lysine), amino acid derivatives (pyroglutamic acid), amines (cadaverine, TMAO, etc.), carbohydrates and vitamins were found as potential markers. The results of the study could be further used for the development of an LC-MS based analytical tool for RJ quality control assessment.

Separation and determination of nimesulide related substances for quality control purposes by micellar electrokinetic chromatography.

A micellar electrokinetic chromatography (MEKC) method has been developed and validated for the determination of nimesulide related compounds in pharmaceutical formulations. Electrophoretic separation of six European Pharmacopoeia (EP) impurities (A-F) was performed using a fused silica capillary (L(eff.)=50 cm, L(tot.)=57 cm, 50 microm i.d.) with a background electrolyte (BGE) containing 25 mM borate buffer (pH 9.5), 30 mM sodium dodecyl sulphate and phi=3% (v/v) acetonitrile. The influence of several factors (surfactant and buffer concentration, pH, organic modifier, applied voltage, capillary temperature and injection time) was studied. The method was suitably validated with respect to linearity, limit of detection and quantification, accuracy, precision and selectivity. The calibration curves obtained for the six compounds were linear over the range 5-12 microgml(-1) (0.05-0.12%). The relative standard deviations (s(r)) of intra- and inter-day experiments were less than 5.0%. The detection limits ranged between 0.7 and 1.6 microgml(-1) depending on the impurity. The proposed method was applied successfully to the quantification of nimesulide impurities in its pharmaceutical formulation.

Development and validation of an ultra high performance liquid chromatography-tandem mass spectrometry method for the determination of phthalate esters in Greek grape marc spirits.

An Ultra High Performance Liquid Chromatography - Tandem Mass Spectrometry method has been developed for the analysis of 12 phthalate esters in Greek grape marc spirits. The phthalates were separated on a U-VDSpher PUR 100 C18-E (100 mm x 2.0 mm, 1.8 µm) column by gradient elution. The analytes were ionized by positive electrospray ionization using the multiple reaction monitoring mode. The standard addition method was used for quantification and the Student's t-test was carried out to evaluate the matrix effect. The accuracy of the method was assessed by recovery experiments resulting in values from 81.6 to 109.6%. The detection limits ranged from 0.3 to 33.3 µg L-1.The proposed method was validated and successfully applied to the analysis of 45 samples collected from Greece and Cyprus. All phthalate esters proved to be present at least once in the analysed grape marc spirits samples, except only in cases of diphenyl phthalate and diisodecyl phthalate, while for the regulated phthalates only bis (2-ethylhexyl) phthalate was quantified above the legislative concentration limits.

[Risk factors for mortality after subarachnoid hemorrhage: a retrospective observational study]. / Fatores de risco para mortalidade após hemorragia subaracnoidea: estudo observacional retrospectivo.

BACKGROUND AND OBJECTIVES: Subarachnoid haemorrhage is an important cause of morbidity and mortality. The aim of the study was to determine predictors of mortality among patients with subarachnoid hemorrhage hospitalized in an Intensive Care Unit. METHODS: This is a retrospective study of patients with subarachnoid hemorrhage admitted to the Intensive Care Unit of our institution during a 7 year period (2009-2015). Data were collected from the Intensive Care Unit computerized database and the patients' chart reviews. RESULTS: We included in the study 107 patients with subarachnoid haemorrhage. A ruptured aneurysm was the cause of subarachnoid haemorrhage in 76 (71%) patients. The overall mortality was 40% (43 patients), and was significantly associated with septic shock, midline shift on CT scan, inter-hospital transfer, aspiration pneumonia and hypernatraemia during the first 72 hours of Intensive Care Unit stay. Multivariate analysis of patients with subarachnoid hemorrhage following an aneurysm rupture revealed that mortality was significantly associated with septic shock and hypernatremia during the first 72 hours of Intensive Care Unit stay, while early treatment of aneurysm (clipping or endovascular coiling) within the first 72 hours was identified as a predictor of a good prognosis. CONCLUSIONS: Transferred patients with subarachnoid haemorrhage had lower survival rates. Septic shock and hypernatraemia were important complications among critically ill patients with subarachnoid haemorrhage and were associated increased mortality.

Optimization and validation of a dissolution test for selegiline hydrochloride tablets by a novel rapid HPLC assay using a monolithic stationary phase.

The present study reports the optimization and validation of a dissolution test for selegiline.HCl tablets using a new high-performance liquid chromatographic (HPLC) method. Rapid separation of the analyte from sample matrix was achieved in less than 60s using a Cromolith RP-18e monolithic column using UV detection at 220 nm. Thorough validation of the assay based on pre-defined criteria included linearity, LOD/LOQ, accuracy, precision, selectivity and ruggedness. The dissolution test was optimized in terms of dissolution medium, basket (type I)/paddle (type II) agitation and rotation speed. Its ruggedness was also validated. The presented analytical and dissolution procedures are currently being applied in the quality and stability control of Cosmopril tablets (5mg/tablet selegiline.HCl, Cosmopharm Ltd., Korinthos, Greece).

Targeted profiling of hydrophilic constituents of royal jelly by hydrophilic interaction liquid chromatography-tandem mass spectrometry.

In the present work a Hydrophilic Interaction Liquid Chromatography-tandem Mass Spectrometry (HILIC-MS/MS) method was developed for the efficient separation and quantification of a large number of small polar bioactive molecules in Royal Jelly. The method was validated and provided satisfactory detection sensitivity for 88 components. Quantification was proven to be precise for 64 components exhibiting good linearity, recoveries R% >90% for the majority of analytes and intra- and inter-day precision from 0.14 to 20% RSD. Analysis of 125 fresh royal jelly samples of Greek origin provided useful information on royal jelly's hydrophilic bioactive components revealing lysine, ribose, proline, melezitose and glutamic acid to be in high abundance. In addition the occurrence of 18 hydrophilic nutrients which have not been reported previously as royal jelly constituents is shown.

Risk factors and predictors of carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter baumannii mortality in critically ill bacteraemic patients over a 6-year period (2010-15): antibiotics do matter.

Purpose. Acinetobacter baumannii and Pseudomonas aeruginosa provoke serious infections, especially in intensive care unit (ICU) patients.Methodology. The risk factors and predictors of mortality for P. aeruginosa (n=84; 46 carbapenem-resistant) and A. baumannii (n=129; all carbapenem-resistant) bloodstream infections (BSIs) in an ICU were evaluated. Antibiotic susceptibility testing was performed using the agar disk diffusion method according to EUCAST guidelines. The minimum inhibitory concentration was determined by a gradient method (Etest). Multilocus sequence typing (MLST) was performed for P. aeruginosa during the carbapenem-resistant outbreak in 2014. Epidemiological data were collected from the patients' chart reviews.Results/Key findings. Hospitalization during the summer months, prior KPC-producing Klebsiella pneumoniae (KPC-Kp) BSI, and the administration of tigecycline, aminoglycosides and cortisone were independently associated with P. aeruginosa BSIs. MLST revealed the dissemination of clone ST227, including carbapenem-resistant P. aeruginosa strains. Hospitalization during the summer months, prior KPC-Kp BSI, and the administration of antibiotics, carbapenem and cortisone were independently associated with A. baumannii BSIs. The 30-day mortality rate for P. aeruginosa and A. baumannii BSI was 45.2 and 39.5 %, respectively. Sequential organ failure assessment (SOFA) score at onset, septic shock, age, and prior KPC-Kp BSI were significantly associated with P. aeruginosa BSI mortality. The administration of at least one active antibiotic was identified as a predictor of a good prognosis. Septic shock and simplified acute physiology score (SAPS) II at onset were independently associated with A. baumannii BSI mortality. The administration of at least one active antibiotic and colistin-vancomycin co-administration were identified as predictors of a good prognosis.Conclusion. KPC-Kp infection predisposes ICU patients to BSI by either A. baumannii or P. aeruginosa. The administration of at least one active antibiotic leads to better survival rates.

Determination of Tyrian purple by high performance liquid chromatography with diode array detection.

Indigotin, indirubin, 6-bromoindigotin, 6'-bromoindirubin, 6-bromoindirubin, 6,6'-dibromoindigotin and 6,6'-dibromoindirubin, the colouring components of Tyrian purple, are quantified by an efficient HPLC method coupled to a diode array detector. The compounds were separated using gradient elution, on a RP-column (Alltima C18, 250mm×3.0mm i.d., 5µm), thermostated at 35°C, with a mobile phase consisting of solvents (Α) H2O+0.1% (v/v) trifluoroacetic acid and (Β) acetonitrile+0.1% (v/v) trifluoroacetic acid, at a flow rate of 0.5mLmin(-1). The method was validated in terms of linearity, detection and quantification limits, precision, accuracy, ruggedness and robustness, the latter with respect to small changes in column temperature and in flow-rate, pH- and solvent composition of the mobile phase. Under optimal conditions, the developed analytical scheme offers limits of detection in the range 0.02-0.05µgmL(-1) and satisfactory linearity up to 2.5µgmL(-1) for all analytes. Four samples produced from the hypobranchial glands of Hexaplex (Murex) trunculus molluscs, collected in the coastlines of Tunisia and Croatia, were treated with hot DMSO and analysed by the established HPLC method, using the standards addition approach. To evaluate the matrix effect, a comparison of the slopes of the standards in solvent regression curves with those of the standard addition method's calibration curves, using the Student's t-test was carried out. The accuracy was evaluated by recovery experiments. Amounts of indigotin, indirubin, and their mono- and dibrominated derivatives ranging between 0.01 to 12.2µgmg(-1) were found in the DMSO extracts of the four molluscan samples.

Preoperative High Dose of Methylprednisolone Improves Early Postoperative Pulmonary Function, in Super-Obese Patients Undergoing Open Surgery; a Prospective, Comparative Study.

BACKGROUND: Surgery stimulates an intense systemic inflammatory response which might increase postoperative morbidity. Corticosteroids may reduce this inflammatory reaction. The purpose of this study was to investigate any possible effect on postoperative morbidity and recovery after administrating methylprednisolone in super-obese patients undergoing open surgery. METHODS: Sixty super-obese patients with BMI ≥50 kg/m2 (mean 57.48 ± 7.33), mean age of 39 ± 9 years, who underwent an open bariatric procedure, were enrolled. Thirty patients (group A) were allocated to a preoperative single dose of 30 mg/kg (ideal body weight) methylprednisolone versus placebo (group B, 30 patients). Endpoints included assessment of IL-6 and CRP; evaluation of postoperative pulmonary function, pain management, nausea, and vomiting; and documentation of postoperative complications. RESULTS: Significant improvement in spirometry parameters and arterial blood gas analysis, in the first and third postoperative days, was observed in the methylprednisolone group. IL-6 and CRP levels were significantly lower in that group. Administration of methylprednisolone was associated with less postoperative pain, nausea, and vomiting, with no statistical difference in septic complications. CONCLUSIONS: Preoperative administration of a single high dose of methylprednisolone in super-obese patients undergoing open surgery inhibits the inflammatory signaling cascade, lessens the systemic inflammatory response, and results in fewer pulmonary complications and better patient recovery.

The Role of Obesity in Sepsis Outcome among Critically Ill Patients: A Retrospective Cohort Analysis.

Background. The objective of this study was to assess the correlation between sepsis, obesity, and mortality of patients admitted to an Intensive Care Unit (ICU). Subjects and Methods. Data of all patients admitted to the ICU of a tertiary hospital during a 28-month period were retrospectively analyzed and included in the study. Results. Of 834 patients included, 163 (19.5%) were obese, while 25 (3.0%) were morbidly obese. Number of comorbidities (P < 0.001), bloodstream infection (P 0.033), and carbapenemase-producing Klebsiella pneumoniae colonization during ICU stay (P 0.005) were significantly associated with obesity, while nonobese patients suffered more frequently from spontaneous intracranial hemorrhage (P 0.038). Total ICU mortality was 22.5%. Increased mortality among obese ICU patients was observed. Sepsis was the main condition of admission for which obese patients had statistically lower survival than normal weight subjects (76.3% versus 43.7%; P 0.001). Mortality of septic patients upon admission was independently associated with SOFA score upon ICU admission (P 0.003), obesity (P 0.014), pneumonia (P 0.038), and development of septic shock (P 0.015). Conclusions. Our study revealed that sepsis upon ICU admission is adversely influenced by obesity but further studies are needed in order to assess the role of obesity in sepsis outcome.

Determination of 1,2-dibromoethane, 1,4-dichlorobenzene and naphthalene residues in honey by gas chromatography--mass spectrometry using purge and trap thermal desorption extraction.

A highly sensitive method for the determination of 1,2-dibromoethane, 1,2-dichlorobenzene and naphthalene residues in honey was developed, using gas chromatography-mass spectrometry combined with a purge and trap thermal desorption system as the extraction technique. Optimal conditions for isolation and separation were established and calibration curves were constructed. Linearity was held between 2.4 and 300 microg kg(-1) honey for 1,2-dibromoethane, 0.5 and 300 microg kg(-1) for 1,4-dichlorobenzene and 0.125 and 3000 microg kg(-1) for naphthalene. The detection limits were found to be 0.8, 0.15 and 0.05 microg kg(-1) honey for 1,2-dibromoethane, 1,4-dichlorobenzene and naphthalene, respectively. The method was applied to the analysis of 25 Greek honey samples. 1,2-Dibromoethane was not found in the majority of the samples, while only one sample was found to contain both 1,4-dichlorobenzene and naphthalene residues at concentrations exceeding 10 microg kg(-1).