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Per- and polyfluoroalkyl substances' (PFAS) extreme persistence has been linked to many adverse effects on human health including increased risk of certain cancers. This study presents the development and validation of a new, highly sensitive method for the quantification of 32 PFAS in human serum using online solid-phase extraction (SPE) coupled with ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Legacy and emerging PFAS were targeted. Main steps of sample pretreatment include protein precipitation (PP), pellet rinsing, centrifugation, preconcentration through solvent evaporation, and online SPE using a weak anion-exchange polymeric sorbent. The PP and pellet-rinsing procedures were optimized through a comprehensive exploration of solvent combinations. Following this, a pretreatment that offers the best compromise for the targeted PFAS was identified using principal component analysis. The method demonstrated excellent linearity (R² = 0.977-0.997) with limits of quantification ranging from 8.9 to 27 ng/L, 5 to 15 times lower than previous methods. Precision (intraday 2.6-14.0 % and interday 1.3-11.0 % relative standard deviation) and accuracy (recoveries 72.7-106 %) were robust. The method was validated in accordance with ISO/IEC 17025 and successfully applied to five human serum samples, confirming its suitability for high-throughput profiling of PFAS in biomonitoring studies. This method is the first to use online SPE for the simultaneous determination of a broad range of PFAS, including ether congeners such as perfluoro(2-ethoxyethane) sulfonic acid and Nafion byproduct 2. Furthermore, control charts were employed to assess instrument performance during routine analysis and implement necessary actions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The intestinal microbiota is composed of a large number of different bacteria that produce a variety of metabolites. Colorectal cancer, which typically develops from adenomatous polyps, is highly influenced by microbiota. Since a variety of molecular changes may occur as these polyps transform from benign tumor to malignant carcinoma, the ability to study the microbiota-produced metabolites can lead to new discoveries about the development and progression of this cancer. However, to address the complexity of the microbiota-produced molecules, novel methods are needed. To this aim, in the present work, we developed a high-throughput metabolomics method to capture the metabolic complexity of the microbiota metabolome adherent to adenomatous polyps and adenocarcinoma. For the first time, the method enables the simultaneous quantification of almost 300 metabolites, while preserving the integrity of the original sample. The metabolomics approach was analytically validated and had excellent performances in terms of recovery, linearity, specificity, intra- and inter-day precision, limits of detection, and quantification. Furthermore, the clinical potential of the method was demonstrated in adenoma collected for a colorectal adenoma study., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The diagnosis and management of Inflammatory bowel disease (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), is still challenging. There is no definitive gold standard diagnostic test, which is made on patient history and with endoscopic and histological findings. This study analyzed serum proteins and fatty acids using mass spectrometry-based techniques. Quantitation of serum proteins was performed by depleting 14 high-abundance proteins, followed by tryptic digestion and LC-MS analysis, while fatty acids were analyzed using GC-MS. Bioinformatic tools were used to identify several new potential biomarkers for an early and non-invasive diagnosis of IBD, and to differentiate CD from UC. Moreover, the diagnostic power of the MS-identified biomarkers was also corroborated by Western Blot and ELISA assays. Hence, we identified the biological functions and pathways involved in the various subsets of IBD. Coagulation, fibrinolysis and acute phase response processes were found to be strongly involved in the condition. The involvement of several fatty acids, such as anti-inflammatory mediators, was also identified. Finally, proteomic and lipidomic data were integrated by using combinatorial and multivariate analyses to discover new combined biomarkers and to study the molecular pathways involved in IBD., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The paper deals with the development of an online UHPLC-MS/MS method for the identification and determination of 25 Endocrine Disrupting Chemicals (EDCs) belonging to different chemical classes (perfluorochemicals, bisphenols, alkylphenols, phthalates, and parabens). The study regards the possible EDC migration from different medical devices of diverse materials used in the routine hospitals for blood sampling or for the parenteral nutrition therapies. In order to evaluate the release of EDCs, the equipment used for blood sampling (syringes, butterfly, cannula-needle, microcapillary tube) was put in contact with a physiological solution at a prefixed time. As regards the migration tests carried out on the medical devices used in the parenteral nutrition therapy (infusion tubes, venous catheters), the catheters were undergone to internal contact with parenteral solution and external contact with physiological one, whereas the infusion tubes were subjected only to internal migration test with parenteral solution. The results demonstrated the released of some ECDs (such as parabens at the concentration of few pg mL -1 , diethylphthalate and dibutylphthlate at ng mL -1, level), when using physiological solution as the leaching agent. In addition, the presence of a PFOS structural isomer was found at 29.0 ng mL -1 : this compound was released in part from the infusion tubes and mostly from the venous catheters, particularly from silicone ones. Using a more lipophilic solution, such as parenteral preparations, a greater amount of EDCs was released, among which also the diethylhexylphthalate that was quantified in all the investigated devices (without highlighting particular trends or migration from PVC devices), but its concentration remained always lower than imposed maximum threshold limit on medical device of 0.1% w/w. The releases from the infusion tubes of the same materials (polyethylene) from different manufacturers are very comparable, whereas the catheter of silicone releases greater amount of EDCs than that of polyurethane., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The authentication and traceability of hazelnuts is very important for both the consumer and the food industry, to safeguard the protected varieties and the food quality. This study investigates the use of a portable FTIR spectrometer coupled to multivariate statistical analysis for the classification of raw hazelnuts. The method discriminates hazelnuts from different origins/cultivars based on differences of the signal intensities of their IR spectra. The multivariate classification methods, namely principal component analysis (PCA) followed by linear discriminant analysis (LDA) and partial least square discriminant analysis (PLS-DA), with or without variable selection, allowed a very good discrimination among the groups, with PLS-DA coupled to variable selection providing the best results. Due to the fast analysis, high sensitivity, simplicity and no sample preparation, the proposed analytical methodology could be successfully used to verify the cultivar of hazelnuts, and the analysis can be performed quickly and directly on site., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Physical activity improves overall health and counteracts metabolic pathologies. Adipose tissue and bone are important key targets of exercise; the prevalence of diseases associated with suboptimal physical activity levels has increased in recent times as a result of lifestyle changes. Mesenchymal stem cells (MSCs) differentiation in either osteogenic or adipogenic lineage is regulated by many factors. Particularly, the expression of master genes such as RUNX2 and PPARγ2 is essential for MSC commitment to osteogenic or adipogenic differentiation, respectively. Besides various positive effects on health, some authors have reported stressful outcomes as a consequence of endurance in physical activity. We looked for further clues about MSCs differentiation and serum proteins modulation studying the effects of half marathon in runners by means of gene expression analyses and a proteomic approach. Our results demonstrated an increase in osteogenic commitment and a reduction in adipogenic commitment of MSCs. In addition, for the first time we have analyzed the proteomic profile changes in runners after half-marathon activity in order to survey the related systemic adjustments. The shotgun proteomic approach, performed through the immuno-depletion of the 14 most abundant serum proteins, allowed the identification of 23 modulated proteins after the half marathon. Interestingly, proteomic data showed the activation of both inflammatory response and detoxification process. Moreover, the involvement of pathways associated to immune response, lipid transport and coagulation, was elicited. Notably, positive and negative effects may be strictly linked. Data are available via ProteomeXchange with identifier PXD006704., Significance: We describe gene expression and proteomic studies aiming to an in-depth understanding of half-marathon effects on bone and adipogenic differentiation as well as biological phenomena involved in sport activity. We believe that this novel approach suggests the physical effects on overall health and show the different pathways involved during half marathon. Contents of the paper have not been published or submitted for publication elsewhere. The authors declare no conflict of interest., (Copyright © 2017 Elsevier B.V. All rights reserved.)

We conducted a proteomics study in order to detect the proteomic method which provides the most complete characterization of the proteins of rice milk. In particular, we compared the results obtained from LC-MS/MS after protein precipitation with acetone or TCA, as well as the results obtained from LC-MS/MS after protein prefractionation based on SDS-PAGE (GeLC-MS/MS) or ProteoMiner™ technology (ProteoMiner-LC-MS/MS), and after peptide prefractionation based on IEF (pIEF-LC-MS/MS). A total of 158 protein species have been detect in rice milk. The physical-chemical analysis and classification of the identified proteins were also reported. In particular, we showed that pIEF-LC-MS/MS method led to a significant increase in the proteome coverage, allowing the identification of a total of 96 proteins of milk rice. This study demonstrates the utility of a prefractionation step based on pIEF before the shotgun proteomic analysis and offers an in-depth insight into the rice milk proteome., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Emerging research has demonstrated that pancreatic ductal adenocarcinoma (PDAC) contains a sub-population of cancer stem cells (CSCs) characterized by self-renewal, anchorage-independent-growth, long-term proliferation and chemoresistance. The secretome analysis of pancreatic CSCs has not yet been performed, although it may provide insight into tumour/microenvironment interactions and intracellular processes, as well as to identify potential biomarkers. To characterize the secreted proteins of pancreatic CSCs, we performed an iTRAQ-based proteomic analysis to compare the secretomes of Panc1 cancer stem-like cells (Panc1 CSCs) and parental cell line. A total of 72 proteins were found up-/down-regulated in the conditioned medium of Panc1 CSCs. The pathway analysis revealed modulation of vital physiological pathways including glycolysis, gluconeogenesis and pentose phosphate. Through ELISA immunoassays we analysed the presence of the three proteins most highly secreted by Panc1 CSCs (ceruloplasmin, galectin-3, and MARCKS) in sera of PDAC patient. ROC curve analysis suggests ceruloplasmin as promising marker for patients negative for CA19-9. Overall, our study provides a systemic secretome analysis of pancreatic CSCs revealing a number of secreted proteins which participate in pathological conditions including cancer differentiation, invasion and metastasis. They may serve as a valuable pool of proteins from which biomarkers and therapeutic targets can be identified., Biological Significance: The secretome of CSCs is a rich reservoir of biomarkers of cancer progression and molecular therapeutic targets, and thus is a topic of great interest for cancer research. The secretome analysis of pancreatic CSCs has not yet been performed. Recently, our group has demonstrated that Panc-1 CSCs isolated from parental cell line by using the CSC selective medium, represent a model of great importance to deepen the understanding of the biology of pancreatic adenocarcinoma. To our knowledge, this is the first proteomic study of pancreatic CSC secretome. We performed an iTRAQ-based analysis to compare the secretomes of Panc1 CSCs and Panc1 parental cell line and identified a total of 43 proteins secreted at higher level by pancreatic cancer stem cells. We found modulation of different vital physiological pathways (such as glycolysis and gluconeogenesis, pentose phosphate pathway) and the involvement of CSC secreted proteins (for example 72kDa type IV collagenase, galectin-3, alpha-actinin-4, and MARCKS) in pathological conditions including cancer differentiation, invasion and metastasis. By ELISA verification we found that MARCKS and ceruloplasmin discriminate between controls and PDAC patients; in addition ROC curve analyses indicate that MARCKS does not have diagnostic accuracy, while ceruloplasmin could be a promising marker only for patients negative for CA19-9. We think that the findings reported in our manuscript advance the understanding of the pathways implicated in tumourigenesis, metastasis and chemoresistance of pancreatic cancer, and also identify a pool of proteins from which novel candidate diagnostic and therapeutic biomarkers could be discovered., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A new approach for monitoring the state of conservation of cultural heritage objects surfaces is being developed. The technique utilizes multi-spectral imaging, multivariate analysis and statistical process control theory for the automatic detection of a possible deterioration process, its localization and identification, and the wavelengths most sensitive to detecting this before the human eye can detect the damage or potential degradation changes occur. A series of virtual degradation analyses were performed on images of parchment in order to test the proposed algorithm in controlled conditions. The spectral image of a Dead Sea Scroll (DSS) parchment, IAA (Israel Antiquities Authority) inventory plate # 279, 4Q501 Apocryphal Lamentations B, taken during the 2008 Pilot of the DSS Digitization Project, was chosen for the simulation., (Copyright © 2011 Elsevier B.V. All rights reserved.)

An on-line solid phase extraction (SPE) ultra high performance liquid chromatography tandem mass spectrometry method has been developed for the simultaneous identification and determination of thirteen polycyclic aromatic hydrocarbons (PAHs) and twelve aldehydes (derivatized with 2,4-dinitrophenylhydrazine). The chromatographic conditions have been optimized to obtain the maximum of sensitivity and resolution taking into account the different retention interactions and the different ionization conditions of PAHs and derivatized aldehydes. LOD values ranging from 0.028 to 0.768 μg L(-1) for PAHs and from 0.002 to 0.125 μg L(-1) for aldehydes were obtained. The resolution permitted the separation of four couples of PAH isomers. Sample pre-treatment and SPE were optimized in order to apply the whole methodology to the analysis of different food matrices as salmon, frankfurter, steak, and pork chop, subjected to different cooking modes (smoked, grilled, cooked in oil or in butter). Particular attention was devoted to the evaluation of matrix effect that was significantly reduced through the on-line SPE treatment. For each food matrix the method detection limits, the method quantitation limits, and the recovery R were evaluated. R was shown not to depend on analyte concentration in the explored concentration range (LOQ - 50.000 μg L(-1)): the average R percent ranges from 70.6% to 120.0%., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A rapid on-line solid phase extraction ultra high performance liquid chromatography tandem mass spectrometry method was developed for the identification and quantitation of nine perfluorinated compounds in matrices of environmental, biological and food interest. Pre-treatment, solid phase extraction, chromatographic and mass detection conditions were optimised, in order to apply the whole methodology to the analysis of different matrices. Particular attention was devoted to the evaluation of matrix effect and the correlated phenomena of ion enhancement or suppression in mass spectrometry detection. LOD and LOQ range from 3 to 15ngL(-1) and from 10 to 50ngL(-1), respectively. Method detection limits (MDLs) were also calculated for each kind of matrix. The recovery, evaluated for each analyte, does not depend on analyte concentration in the explored concentration range: average R¯% values are always greater than 82.9%. In the whole, the results obtained for samples of river waters, blood serum, blood plasma, and fish confirm the ubiquitous presence of perfluorinated compounds, as recently denounced by many sources., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Surgical reconstruction of the oral commissure aims to restore both symmetry of the lips at rest and, more importantly, full oral competence. Moulding the lip commissure with functional and cosmetic fidelity remains till today a difficult task. A possible surgical solution, the 'elastic flap' principle described by Goldstein, may be found in the wide full-thickness mobilization of the upper and lower vermilion as two composite myocutaneous flaps - tissue sandwiches consisting of labial skin, orbicularis oris muscle and oral mucosa - on the axial pattern of the superior and inferior labial arteries. Based on the contralateral commissure, both flaps are easily 'stretched', accordion-like, to reach the predetermined point of the new commissure, using to full advantage the inherent elastic potential of both vermilions. The fibres of the orbicularis oris muscle at each end of both flaps are embricated to reconstitute a neo-modiolus, which is anchored to the residual buccinator muscle in primary reconstructions, or to the available peri-oral fibrous tissue in secondary procedures. The authors present a select group of 22 patients, who, between 1993 and 2008, underwent this reconstruction procedure for primary or secondary defects involving the oral commissure. The results were generally satisfactory, both functionally and cosmetically. The advantages of this procedure include full restoration of the dynamic function of the orbicularis ring in a single-stage operation and avoidance of either lipswitching procedures or of mobilization of mucosa and cheek skin. The final scars remain well camouflaged within the oral mucosa and the mucocutaneous junction of each lip., ((c) 2008 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

This paper reports the development of calibration models for quality control in the production of ethylene/propylene/1-butene terpolymers by the use of multivariate tools and FT-IR spectroscopy. 1-Butene concentration prediction is achieved in terpolymers by coupling FT-IR spectroscopy to multivariate regression tools. A dataset of 26 terpolymers (14 coming from a constrained experimental design for mixtures, plus 12 terpolymers used for external validation) was analysed by FT-IR spectroscopy. An internal method of "Polimeri Europa" plant, based on (13)C NMR spectroscopy is used to determine the percentage of 1-butene in the samples. Then, different multivariate tools are used for 1-butene concentration prediction based on the FT-IR spectra recorded. Different multivariate calibration methods were explored: principal component regression (PCR), partial least squares (PLS), stepwise OLS regression (SWR) and artificial neural networks (ANNs). The model obtained by back-propagation neural networks turned out to be the best one. The performances of the BP-ANN model were further improved by variable selection procedures based on the calculation of the first derivative of the network. The proposed approach allows the monitoring in real time of the polymer synthesis and the estimation of the characteristics of the product attainable from the concentration of 1-butene.

The optimisation of the sensitivity in the ICP-MS determination of 83 isotopes, as a function of 21 operative parameters was performed by generating an initial experimental design that was used to define, by principal component analysis, the multi-criteria target function. The first PC, which contained an overall evaluation of the signal intensity of all isotopes, was used to rank the experiments. The modified simplex optimisation technique was then applied on the ranked experiments. The increase in signal intensity was, on the average, 3.9 times for the isotopes considered for the simplex procedure. When finally convergence was achieved, a PLS regression model calculated on the available experiments allowed to investigate the effect played by each factor on the experimental response. Simplex and PCA proved to be extremely effective to obtain the optimisation and to generate the multi-criteria target function: they can be suggested as an automatic method to perform the optimisation of the instrumental operative conditions.

A combined microvascular flap composed of serratus anterior myo-osseous and a latissimus dorsi myocutaneous flap has been performed for resurfacing massive scalp and skull defects, accompanied by chronic infection and heavy radiation damage. The authors present a case report where the combined procedure allowed a single-stage reconstruction of this complex defect.

In this paper, Legendre moments are calculated to extract the global information from a set of two-dimensional polyacrylamide gel electrophoresis map images. The dataset contains 18 samples belonging to two different cell lines (PACA44 and T3M4) of control (untreated) and drug-treated pancreatic ductal carcinoma cells. The aim of this work was to obtain the correct classification of the 18 samples, using the Legendre moments as discriminant variables. For each image the Legendre moments up to a maximum order of 100 were computed. The stepwise linear discriminant analysis (LDA) was performed in order to select the moments with the highest discriminating power. The results demonstrate that the Legendre moments can be successfully applied for fast classification purposes and similarity analysis.

This work is an extension of a method for monitoring the conservation state of pigmented surfaces presented in a previous paper. A cotton canvas painted with an organic pigment (Alizarin) was exposed to UV light in order to evaluate the effects of the applied treatment on the surface of the sample. The conservation state of the pigmented surface was monitored with ATR-FT-IR spectroscopy and multivariate control charts. The IR spectra were analysed by principal component analysis (PCA) and the relevant principal components (PCs) were used for constructing multivariate Shewhart, cumulative sums (CUSUM) and simultaneous scores monitoring and residuals tracking (SMART) control charts. These tools were successfully applied for the identification of the presence of relevant modifications occurring on the surface of the sample. Finally, with the aim to more deeply investigate what happened to the sample surface during the UV exposure, a PCA of the residuals matrix of degradation analyses only, not present in the previous paper, was performed. This analysis produced interesting results concerning the identification of the processes taking place on the irradiated surface.

Two of the most suitable analytical techniques used in the field of cultural heritage are NIR (near-infrared) and Raman spectroscopy. FT-Raman spectroscopy coupled to multivariate control charts is applied here for the development of a new method for monitoring the conservation state of pigmented and wooden surfaces. These materials were exposed to different accelerated ageing processes in order to evaluate the effect of the applied treatments on the goods surfaces. In this work, a new approach based on the principles of statistical process control (SPC) to the monitoring of cultural heritage, has been developed: the conservation state of samples simulating works-of-art has been treated like an industrial process, monitored with multivariate control charts, owing to the complexity of the spectroscopic data collected. The Raman spectra were analysed by principal component analysis (PCA) and the relevant principal components (PCs) were used for constructing multivariate Shewhart and cumulative sum (CUSUM) control charts. These tools were successfully applied for the identification of the presence of relevant modifications occurring on the surfaces. CUSUM charts however proved to be more effective in the identification of the exact beginning of the applied treatment. In the case of wooden boards, where a sufficient number of PCs were available, simultaneous scores monitoring and residuals tracking (SMART) charts were also investigated. The exposure to a basic attack and to high temperatures produced deep changes on the wooden samples, clearly identified by the multivariate Shewhart, CUSUM and SMART charts. A change on the pigment surface was detected after exposure to an acidic solution and to the UV light, while no effect was identified on the painted surface after the exposure to natural atmospheric events.

Background: The adrenal gland is the election organ forming primary neuroblastoma (NB) tumours, the most common extracranial solid tumours of infancy and childhood., Methods: Samples of adrenal gland belonging to healthy and diseased nude mouse were analysed by 2D gel-electrophoresis. The resulting 2D-PAGE maps were digitized by PDQuest and investigated by principal component analysis (PCA)., Results: The analysis of the loadings of the first principal component (PC) permitted the evaluation of the spots characterising each class of samples. Moreover, the soft-independent model of class analogy (SIMCA) method confirmed the separation of the samples in the two classes and allowed the identification of the modelling and discriminating spots. Very good correlation was found between the data obtained by analysis of 2D maps via the commercial software PDQuest and the present PCA analysis. In both cases, the comparison between such maps showed up- and down-regulation of 84 polypeptide chains, out of a total of 700 spots detected by a fluorescent stain, Sypro Ruby. Spots that were differentially expressed between the two groups were analysed by matrix-assisted laser desorption time-of-flight (MALDI-TOF) mass spectrometry and 14 of these spots were identified so far., (Copyright 2004 Elsevier B.V.)

Two-dimensional (2D) electrophoresis is the most wide spread technique for the separation of proteins in biological systems. This technique produces 2D maps of high complexity, which creates difficulties in the comparison of different samples. The method proposed in this paper for the comparison of different 2D maps can be summarised in four steps: (a) digitalisation of the image; (b) fuzzyfication of the digitalised map in order to consider the variability of the two-dimensional electrophoretic separation; (c) decoding by principal component analysis of the previously obtained fuzzy maps, in order to reduce the system dimensionality; (d) classification analysis (linear discriminant analysis), in order to separate the samples contained in the dataset according to the classes present in said dataset. This method was applied to a dataset constituted by eight samples: four belonging to healthy human lymph-nodes and four deriving from non-Hodgkin lymphomas. The amount of fuzzyfication of the original map is governed by the sigma parameter. The larger the value, the more fuzzy theresulting transformed map. The effect of the fuzzyfication parameter was investigated, the optimal results being obtained for sigma = 1.75 and 2.25. Principal component analysis and linear discriminant analysis allowed the separation of the two classes of samples without any misclassification.

The instep of the foot is an excellent donor site for split-thickness skin grafts to cover soft tissue defects of the palm and digits. The technique is described and three illustrative clinical cases are presented. The unique, specialized characteristics of plantar skin are described, emphasizing similarities to palmar skin. The indications for this technique are hyperpigmentation, hair growth, ulceration, hyperkeratosis, marginal scarring, or recurrent breakdown of a preexisting graft, but we have also employed it for primary reconstructions in selected circumstances. The instep graft provides ideal color and texture match and long durability for a palmar graft, with the added advantage of an inconspicuous donor site.