Parameter space exploration within dynamic simulations of signaling networks

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Residue analysis of cortisonics in bovine milk by LC tandem MS

A new method of analysis, based on liquid chromatography-tandem mass spectrometry, was developed for the simultaneous determination of twelve steroidal anti-inflammatory drugs in cow milk. This analytical method was validated according to the European Commission Decision 2002/657/EC. Analytes were separated by ion-suppression reversed-phase liquid chromatography, due to weakly acidic nature of the selected cortisonics. Dexamethasone tetradeuterated was chosen as internal standard. Detection was in positive ion mode, using a high flow electrospray interface (TurboIonSpray). Two Multi Reaction Monitoring (MRM) transitions each analyte were selected. Analytes’ recovery in milk was made up of two steps: 1) sample deproteinization by trifluoroacetic acid; 2) solid phase extraction (SPE) by means of C18 as adsorbent and methanol as eluent. Recoveries were ≥ 70% with relative standard deviations minor than 10%. Matrix effect has become unimportant owing to the efficacy of the extraction procedure in interfering substances removal and to a suitable internal standard. In order to evaluate matrix effect, calibration curves in solvent were compared with calibration curves obtained by blank extracts spiked with analytes. Method limits were evaluated on the quantifier transition (less intense), according to the European Commission Decision 2002/657/EC for decision limit (CCalfa) and detection capability (CCbeta) assessment. Monitoring results on different fresh whole milk brands, have shown in some cases a suspected positivity. Hydrocortisone and Cortisone Acetate were found in these samples at levels between CCalfa and CCbeta. This LC-MS/MS study allowed to observe Triamcinolone transformation during the evaporation step, that could explain low recovery data found in literature

Residue analysis of glucocorticoids in bovine milk by liquid chromatography-tandem mass spectrometry

A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of 13 steroidal anti-inflammatory drugs in bovine milk is presented. Due to their weakly acid nature, analytes were separated by ion suppression reversed phase chromatography and detected in positive-ion mode by a high flow electrospray source. Dexamethasone-d4 was used as internal standard. The sample preparation was simple and reliable; it included acidic deproteinization of milk followed by sample enrichment and clean-up, utilizing a C18 solid phase extraction cartridge. Recoveries exceeded 70% with an intra-day precision not larger than 12%. The efficiency of the sample clean-up and internal standardization rendered negligible the matrix effect, estimated by comparing standard and matrix-matched calibration curves. A small-scale reconnaissance was carried out on several raw and whole fresh milk samples. A large number of analyzed samples showed a chromatographic peak, in the retention time window of cortisol, at levels included between its decision limit (CC alpha) and detection capability (CC beta). As a result of a heat-induced transformation, an isomeric product of triamcinolone was observed during the extract evaporation. Since this rearrangement might occur during the milk pasteurization process, LC-MS/MS and (1)H-NMR investigations were performed out to conclusively differentiate the two isomers. One- and two-dimensional proton NMR spectra were able to identify the transformation product as 9a-fluoro-11b,16a-trihydroxy-17b-hydroxymethyl-D-homoandrosta-1,4-diene-3,17a-dione

Assessment of Disease Activity in Small Bowel Crohn’s Disease: Comparison between Endoscopy and Magnetic Resonance Enterography Using MRIA and Modified MRIA Score

Objectives. To retrospectively compare the results of the MRIA (magnetic resonance index of activity) with a modified MRIA (mMRIA), which was calculated excluding from MRIA formula the data of relative contrast enhancement (RCE). Materials and Methods. MR-E and corresponding endoscopic records of 100 patients were reviewed. MRIA, mMRIA, and SES endoscopic index were calculated for all the patients. Namely, MRIA was calculated as follows: (1.5 × wall thickening + 0.02 × RCE + 5 × intramural edema + 10 × ulcers), while mMRIA was calculated with the modified formula (1.5 × wall thickening + 5 × intramural edema + 10 × ulcers). Results. Mean MRIA and mMRIA values were 19.3 and 17.68, respectively p < 0.0001. A significant correlation p < 0.0001 was observed between MRIA and mMRIA scores and between both MR indexes and SES p < 0.0001. Conclusions. mMRIA was comparable to MRIA in the evaluation of disease activity in Crohn's disease

The Italian version of the Reflective Functioning Questionnaire: Validity data for adults and its association with severity of borderline personality disorder.

INTRODUCTION:Impairments in the ability to understand others and the self in terms of internal mental states (reflective functioning [RF] or mentalizing) are thought to play a key role in the development of borderline personality disorder (BPD). The first aim of this study was to validate the Italian version of the Reflective Functioning Questionnaire (RFQ), a brief self-report measure of RF, by examining its factor structure with Principal Component Analyses (PCA), and correlations with constructs that should be theoretically related to RF. In addition, we investigated whether the RFQ could empirically distinguish between healthy controls and carefully diagnosed BPD patients using Research Operating Curve methods, and was related to severity of borderline pathology as measured with the Shedler-Westen Assessment Procedure (SWAP), an observer-rated measure of BPD pathology. METHODS:An Italian translation of the RFQ was administered to a sample of 154 healthy controls and a clinical sample of 59 BPD patients diagnosed with the Structured Clinical Interview for DSM-IV Axis II disorders. Clinical severity of BPD was assessed with the SWAP. Normal controls completed self-report inventories of constructs related to RF (mindfulness, empathy, Theory of Mind, alexithymia, and autistic traits). RESULTS:PCA confirmed the a priori factor structure in the Italian translation of the RFQ, showing two subscales that measure certainty and uncertainty about mental states, with satisfactory reliability and construct validity. These dimensions also distinguished BPD patients from healthy controls (p < 0.05). ROC analyses showed that the uncertainty subscale discriminated BPD patients from healthy individuals (area under the curve = 78%, cut of 4.5 points, sensitivity = 73%, specificity = 68%). Within the patient group, regression analyses showed uncertainty about mental states to have a significant unique contribution in predicting BPD severity (p < 0.05), explaining 12% of the variance. CONCLUSIONS:Results largely supported the reliability and validity of the Italian version of the RFQ. These findings also provide further evidence for the role of impairments in mentalizing and reinforce the rationale for offering mentalization-based interventions to individuals with this disorder