Cross-contamination of eukaryotic cell lines used in biomedical research represents a highly relevant problem. Analysis of repetitive DNA sequences, such as Short Tandem Repeats (STR), or Simple Sequence Repeats (SSR), is a widely accepted, simple, and commercially available technique to authenticate cell lines. However, it provides only qualitative information that depends on the extent of reference databases for interpretation. In this work, we developed and validated a rapid and routinely applicable method for evaluation of cell culture cross-contamination levels based on mass spectrometric fingerprints of intact mammalian cells coupled with artificial neural networks (ANNs). We used human embryonic stem cells (hESCs) contaminated by either mouse embryonic stem cells (mESCs) or mouse embryonic fibroblasts (MEFs) as a model. We determined the contamination level using a mass spectra database of known calibration mixtures that served as training input for an ANN. The ANN was then capable of correct quantification of the level of contamination of hESCs by mESCs or MEFs. We demonstrate that MS analysis, when linked to proper mathematical instruments, is a tangible tool for unraveling and quantifying heterogeneity in cell cultures. The analysis is applicable in routine scenarios for cell authentication and/or cell phenotyping in general.

• MeSH
• Principal Component Analysis MeSH
• Cell Line MeSH
• Mass Spectrometry methods   MeSH
• Calibration MeSH
• Coculture Techniques MeSH
• Humans MeSH
• Human Embryonic Stem Cells physiology   MeSH
• Multivariate Analysis MeSH
• Mice MeSH
• Neural Networks, Computer * MeSH
• Specimen Handling MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Kinetocardiography MeSH
• Coronary Disease diagnosis   MeSH
• Mathematics MeSH

Visible near-infrared (Vis-NIR) reflection spectroscopy and mid-infrared (mid-IR) reflection spectroscopy are cost- and time-effective and environmentally friendly techniques that could be alternatives to conventional soil analysis methods. Successful determination of spectrally active soil components, including soil organic matter (SOM), depends on the selection of suitable pretreatment and multivariate calibration techniques. The objective of the present review is to critically examine the suitability of Vis-NIR (350-2500 nm) and mid-IR (4000-400 cm(-1)) spectroscopy as a tool for SOM quantity and quality determination. Particular attention is paid to different pretreatment and calibration procedures and methods, and their ability to predict SOM content from Vis-NIR and mid-IR data is discussed. We then review the most recent research using spectroscopy in different calibration scales (local, regional, or global). Finally, accuracy and robustness, as well as uncertainty in Vis-NIR and mid-IR spectroscopy, are considered. We conclude that spectroscopy, especially the mid-IR technique in association with Savitzky-Golay smoothing and derivatization and the least squares support vector machine (LS-SVM) algorithm, can be useful in determining SOM quantity and quality. Future research conducted for the standardization of protocols and soil conditions will allow more accurate and reliable results on a global and international scale.

• MeSH
• Spectroscopy, Near-Infrared methods   standards   MeSH
• Calibration MeSH
• Least-Squares Analysis MeSH
• Soil chemistry   MeSH
• Spectrophotometry, Infrared methods   standards   MeSH
• Support Vector Machine MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

BACKGROUND AND PURPOSE: ASTRAL (Acute Stroke Registry and Analysis of Lausanne) and DRAGON (includes dense middle cerebral artery sign, prestroke modified Rankin Scale score, age, glucose, onset to treatment, National Institutes of Health Stroke Scale score) are 2 recently developed scores for predicting functional outcome after acute stroke in unselected acute ischemic stroke patients and in patients treated with intravenous thrombolysis, respectively. We aimed to perform external validation of these scores to assess their predictive performance in the large multicentre Safe Implementation of Thrombolysis in Stroke-International Stroke Thrombolysis Register. METHODS: We calculated the ASTRAL and DRAGON scores in 36 131 and 33 716 patients, respectively, registered in Safe Implementation of Thrombolysis in Stroke-International Stroke Thrombolysis Register between 2003 and 2013. The proportion of patients with 3-month modified Rankin Scale scores of 3 to 6 was observed for each score point and compared with the predicted proportion according to the risk scores. Calibration was assessed using calibration plots, and predictive performance was assessed using area under the curve of the receiver operating characteristic. Multivariate logistic regression coefficients for the variables in the 2 scores were compared with the original derivation cohorts. RESULTS: The ASTRAL showed an area under the curve of 0.790 (95% confidence interval, 0.786-0.795) and the DRAGON an area under the curve of 0.774 (95% confidence interval, 0.769-0.779). All ASTRAL parameters except range of visual fields and all DRAGON parameters were significantly associated with functional outcome in multivariate analysis. CONCLUSIONS: The ASTRAL and DRAGON scores show an acceptable predictive performance. ASTRAL does not require imaging-data and therefore may have an advantage for the use in prehospital patient assessment. Prospective studies of both scores evaluating the impact of their use on patient outcomes after intravenous thrombolysis and endovascular therapy are needed.

• MeSH
• Algorithms * MeSH
• Stroke physiopathology   therapy   MeSH
• Brain Ischemia physiopathology   therapy   MeSH
• Calibration MeSH
• Middle Aged MeSH
• Humans MeSH
• Recovery of Function * MeSH
• Area Under Curve MeSH
• Predictive Value of Tests MeSH
• Prospective Studies MeSH
• Reference Values MeSH
• Registries MeSH
• Reproducibility of Results MeSH
• ROC Curve MeSH
• Aged MeSH
• Thrombolytic Therapy MeSH
• Treatment Outcome MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Multicenter Study MeSH
• Validation Study MeSH

A capillary electrophoresis method for the simultaneous determination of the enantiomeric purity and of impurities of the chiral drug ambrisentan has been developed following the Quality by Design principles. The selected separation system consisted of a micellar pseudostationary phase made by sodium dodecyl sulphate with the addition of γ-cyclodextrin. The effects of critical process parameters (capillary length, temperature, voltage, borate concentration, pH, sodium dodecyl sulphate concentration, γ-cyclodextrin concentration) on enantioresolution of ambrisentan and analysis time were extensively investigated by multivariate strategies involving a screening phase and Response Surface Methodology. The Design Space was defined with a desired probability level π≥90%, and the working conditions, with the limits of the Design Space, corresponded to the following: capillary length, 64.5cm; temperature, 22°C; voltage, 30kV (26-30kV); background electrolyte, 100mM borate buffer pH 9.20 (8.80-9.60), 100mM sodium dodecyl sulphate, 50mM (43-50mM) γ-cyclodextrin. A Plackett-Burman design was applied for robustness testing, and a method control strategy was established. The method was fully validated according to the International Conference on Harmonisation guidelines and was applied to ambrisentan coated tablets.

• MeSH
• Borates chemistry   MeSH
• Chromatography, Micellar Electrokinetic Capillary * methods   MeSH
• Cyclodextrins MeSH
• Sodium Dodecyl Sulfate MeSH
• Phenylpropionates analysis   chemistry   MeSH
• Calibration MeSH
• Hydrogen-Ion Concentration MeSH
• Drug Contamination MeSH
• Buffers MeSH
• Pyridazines analysis   chemistry   MeSH
• Reproducibility of Results MeSH
• Quality Control MeSH
• Stereoisomerism MeSH
• Tablets analysis   MeSH
• Publication type
• Journal Article MeSH

To compare the predictivity of the neuroactive steroids in the cerebrospinal fluid and peripheral blood for the diagnostics of CNS disturbances, eighteen unconjugated steroids were quantified in the cerebrospinal fluid (CSF) from the 3rd ventricle and 18 unconjugated steroids and 7 steroid polar conjugates were measured in the serum using GC-MS and RIA. Eight postmenopausal women (56-78 years of age) and 7 men (22-88 years of age) with hydrocephalus were enrolled in the study. The sensitivity of the method ranged from low femtogram to low picogram levels depending on the steroid fragmentation pattern. Using multivariate regression, a model for simultaneous prediction of the CSF steroids from the serum steroids was completed. Then, the penetrability of the individual steroids across the blood-brain-barrier was evaluated and the sources of various brain steroids were estimated. Our data show that a part of the steroids may be synthesized de novo in the CNS. However, substantial part of the steroid metabolites may be synthesized in the CNS from the steroid precursors or directly transported from the periphery. The CNS in situ synthesis and transport from periphery might be complementary in some cases, i.e. brain synthesis might provide minimum level of steroids, which are indispensable for the CNS functions. Copyright 2010 Elsevier Ltd. All rights reserved.

• MeSH
• Diagnostic Techniques, Neurological standards   MeSH
• Adult MeSH
• Hydrocephalus blood   MeSH
• Calibration MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Central Nervous System Diseases diagnosis   blood   MeSH
• Gas Chromatography-Mass Spectrometry methods   standards   MeSH
• Postmenopause blood   MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Sensitivity and Specificity MeSH
• Steroids blood   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

The ultrahigh-performance liquid chromatography-mass spectrometry (UHPLC/MS) method was optimized and validated for the determination of oxylipins in human plasma using the targeted approach with selected reaction monitoring (SRM) in the negative-ion electrospray ionization (ESI) mode. Reversed phase UHPLC separation on an octadecylsilica column enabled the analysis of 63 oxylipins including numerous isomeric species within 12-min run time. The method was validated (calibration curve, linearity, limit of detection, limit of quantification, carry-over, precision, accuracy, recovery rate, and matrix effect) and applied to 40 human female plasma samples from breast cancer patients and age-matched healthy volunteers (control). Thirty-six oxylipins were detected in human plasma with concentrations above the limit of detection, and 21 of them were quantified with concentrations above the limit of quantitation. The concentrations determined in healthy controls are in a good agreement with previously reported data on human plasma. Quantitative data were statistically evaluated by multivariate data analysis (MDA) methods including principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA). S-plot and box plots showed that 13-HODE, 9-HODE, 13-HOTrE, 9-HOTrE, and 12-HHTrE were the most upregulated oxylipin species in plasma of breast cancer patients.

• MeSH
• Principal Component Analysis MeSH
• Chromatography, Reverse-Phase methods   MeSH
• Spectrometry, Mass, Electrospray Ionization methods   MeSH
• Humans MeSH
• Limit of Detection MeSH
• Multivariate Analysis MeSH
• Breast Neoplasms blood   MeSH
• Oxylipins blood   MeSH
• Reproducibility of Results MeSH
• Chromatography, High Pressure Liquid methods   MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Validation Study MeSH

• MeSH
• Computer Systems MeSH
• Statistics as Topic MeSH

• Conspectus
• Statistika
• NML Fields
• statistika, zdravotnická statistika
• lékařská informatika

• MeSH
• Clinical Medicine MeSH
• Statistics as Topic MeSH
• Publication type
• Abstracts MeSH
• Congress MeSH
• Collected Work MeSH
• News MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• statistika, zdravotnická statistika
• lékařství

BACKGROUND: Prediction of side-specific extraprostatic extension (EPE) is crucial in selecting patients for nerve-sparing radical prostatectomy (RP). Multiple nomograms, which include magnetic resonance imaging (MRI) information, are available predict side-specific EPE. It is crucial that the accuracy of these nomograms is assessed with external validation to ensure they can be used in clinical practice to support medical decision-making. METHODS: Data of prostate cancer (PCa) patients that underwent robot-assisted RP (RARP) from 2017 to 2021 at four European tertiary referral centers were collected retrospectively. Four previously developed nomograms for the prediction of side-specific EPE were identified and externally validated. Discrimination (area under the curve [AUC]), calibration and net benefit of four nomograms were assessed. To assess the strongest predictor among the MRI features included in all nomograms, we evaluated their association with side-specific EPE using multivariate regression analysis and Akaike Information Criterion (AIC). RESULTS: This study involved 773 patients with a total of 1546 prostate lobes. EPE was found in 338 (22%) lobes. The AUCs of the models predicting EPE ranged from 72.2% (95% CI 69.1-72.3%) (Wibmer) to 75.5% (95% CI 72.5-78.5%) (Nyarangi-Dix). The nomogram with the highest AUC varied across the cohorts. The Soeterik, Nyarangi-Dix, and Martini nomograms demonstrated fair to good calibration for clinically most relevant thresholds between 5 and 30%. In contrast, the Wibmer nomogram showed substantial overestimation of EPE risk for thresholds above 25%. The Nyarangi-Dix nomogram demonstrated a higher net benefit for risk thresholds between 20 and 30% when compared to the other three nomograms. Of all MRI features, the European Society of Urogenital Radiology score and tumor capsule contact length showed the highest AUCs and lowest AIC. CONCLUSION: The Nyarangi-Dix, Martini and Soeterik nomograms resulted in accurate EPE prediction and are therefore suitable to support medical decision-making.

• MeSH
• Middle Aged MeSH
• Humans MeSH
• Magnetic Resonance Imaging * methods   MeSH
• Prostatic Neoplasms * diagnostic imaging   pathology   surgery   MeSH
• Nomograms * MeSH
• Prognosis MeSH
• Prostate diagnostic imaging   pathology   surgery   MeSH
• Prostatectomy * methods   MeSH
• Retrospective Studies MeSH
• Robotic Surgical Procedures methods   MeSH
• Aged MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Publication type
• Journal Article MeSH
• Validation Study MeSH