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Liquid chromatography high-resolution mass spectrometry fingerprinting together with pattern recognition techniques was used to determine the metabolites involved in the susceptibility of apple cultivars to rosy apple aphid (RAA). Preprocessing of ultra-high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry raw data of resistant and susceptible apple cultivars was carried out with XCMS and CAMERA packages. Univariate statistical tools and multivariate data analysis highlighted significant different profiles of the apple metabolomes according to their tolerance to RAA. Optimized and cross-validated Partial least squares discriminant analysis and orthogonal projections to latent structures discriminant analysis models confirmed

In hormone-responsive breast cancer cells, progesterone (P4) has been shown to act via its nuclear receptor (PR), a ligand-activated transcription factor. A small fraction of PR is palmitoylated and anchored to the cell membrane (mbPR) forming a complex with estrogen receptor alpha (ERα). Upon hormone exposure, either directly or via interaction with ERα mbPR activates the SRC/RAS/ERK kinase pathway leading to phosphorylation of PR by ERK. Kinase activation is essential for P4 gene regulation, as the ERK and MSK1 kinases are recruited by the PR to its genomic binding sites and trigger chromatin remodeling. An interesting open question is whether activation of mbPR can result in gene regulation in the absence of ligand binding to intracellular PR. This matter has been investigated in the past using P4 attached to serum albumin, but the attachment is leaky and albumin can be endocytosed and degraded, liberating P4. Here we propose a more stringent approach to address this issue by ensuring attachment of P4 to the cell membrane via covalent binding to a stable phospholipid. This strategy identifies the actions of P4 independent of hormone binding to intracellular PR. We found that a membrane-attached progestin can activate mbPR, the ERK signalling pathway leading to PR phosphorylation, initial gene regulation and entry into the cell cycle, in the absence of detectable intracellular progestin.

Lipid imaging mass spectrometry (LIMS) has been tested in several pathological contexts, demonstrating its ability to segregate and isolate lipid signatures in complex tissues, thanks to the technique’s spatial resolution. However, it cannot yet compete with the superior identification power of high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS), and therefore, very often, the latter is used to refine the assignment of the species detected by LIMS. Also, it is not clear if the differences in sensitivity and spatial resolution between the two techniques lead to a similar panel of biomarkers for a given disease. Here, we explore the capabilities of LIMS and HPLC-MS to produce a panel of lipid biomarkers to screen nephrectomy samples from 40 clear cell renal cell carcinoma patients. The same set of samples was explored by both techniques, and despite the important differences between them in terms of the number of detected and identified species (148 by LIMS and 344 by HPLC-MS in negative-ion mode) and the presence/absence of image capabilities, similar conclusions were reached: using the lipid fingerprint, it is possible to set up classifiers that correctly identify the samples as either healthy or tumor samples. The spatial resolution of LIMS enables extraction of additional information, such as the existence of necrotic areas or the existence of different tumor cell populations, but such information does not seem determinant for the correct classification of the samples, or it may be somehow compensated by the higher analytical power of HPLC-MS. Similar conclusions were reached with two very different techniques, validating their use for the discovery of lipid biomarkers. The work was funded by the Basque Government (IT971-16, IT1162-19, and ELKARTEK KK2018-00090) and has been developed as a Ph.D. project of LMS, who is the recipient of a Predoctoral Fellowship from the Spanish Government (BES- 2016-078721) . The authors are grateful to SGiker Lipidomic Service (UPV/EHU, MICINN, GV/EG, ESF) for the expert advice and technical and human support in MALDI and HPLC- MS analysis.

Love is a worldwide known phenomenon that affects many aspects of human life, including considering a romantic partner with whom to bond. Thus, developing a reliable and valid measure of love experiences is crucial. One of the most popular tools to test love levels is Sternberg's 45-item Triangular Love Scale (TLS-45), which measures three love components: Intimacy, Passion, and Commitment. However, our literature review reveals that the majority of studies (64%) use a broad variety of shortened versions of the TLS-45. To achieve scientific consensus and improve the reliability, comparability, and generalizability of results across studies, here, we develop a short version of the scale–the TLS-15–which contains 15 items with a 5-point rather than 9-point response scale. In Study 1 (N = 7,332), we re-analyzed secondary data from a large-scale multinational study that validated the original TLS-45 to establish whether the scale could be truncated. In Study 2 (N = 307), we provided evidence for the three-factor structure of the TLS-15 and its reliability. Study 3 (N = 413) confirmed convergent validity and test-retest stability of the TLS-15. Study 4 (N = 60,311) presented a large-scale validation across 37 linguistic versions of the TLS-15 on a cross-cultural sample spanning every continent of the globe. The overall results provided support for the reliability, validity, and cross-cultural invariance of the TLS-15, which could be used as a measure of love components–separately or jointly as a three-factor measure.

Recent technical advances in mass spectrometry, as applied to the analytical chemistry of lipid molecules, enable the simultaneous detection of the multiplicity of lipid complex species present in the human brain. This, in combination with quantitative studies carried out in plasma samples, helps to identify disease biomarkers including for Alzheimer's disease (AD). Mass spectrometry imaging (MSI) is particularly powerful for the anatomical localization of lipids in brain slices, identifying lipid modifications in postmortem frozen samples from AD patients.Human brain tissues are sectioned in a cryostat and then covered with a chemical matrix, such as mercaptobenzothiazole (MBT) or α-cyano-4-hydroxycinnamic acid (CHCA), to ionize the lipid molecules either by sublimation or by spraying. We describe the use of matrix-assisted laser desorption ionization (MALDI) in an LTQ-Orbitrap-XL mass spectrometer to scan brain tissue slices with high spatial resolution, analyzing 50 μm cell layers. The lipid spectra obtained for each pixel are transformed to color-coded intensity maps of hundreds of lipid species included those within a single tissue slice.

Background: Abnormal cholesterol metabolism changes the neuronal membrane and may promote amyloidogenesis. Oxysterols in cerebrospinal fluid (CSF) are related to Alzheimer's disease (AD) biomarkers in mild cognitive impairment and dementia. Cholesterol turnover is important for axonal and white matter (WM) microstructure maintenance. Objective: We aim to demonstrate that the association of oxysterols, AD biomarkers, and WM microstructure occurs early in asymptomatic individuals. Methods: We studied the association of inter-individual variability of CSF 24-hydroxycholesterol (24-OHC), 27-hydroxycholesterol (27-OHC), 7-ketocholesterol (7-KC), 7 beta-hydroxycholesterol (7 beta-OHC), amyloid-beta(42) (A beta(42)), total-tau (t-tau), phosphorylated-tau (p-tau), neurofilament (NfL), and WM microstructure using diffusion tensor imaging, generalized linear models and moderation/mediation analyses in 153 healthy adults. Results: Higher 7-KC levels were related to lower A beta(42), indicative of greater AD pathology (p = 0.041). Higher 7-KC levels were related to lower fractional anisotropy (FA) and higher mean (MD), axial (AxD), and radial (RD) diffusivity. 7-KC modulated the association between AxD and NfL in the corpus callosum splenium (B = 39.39, p = 0.017), genu (B = 68.64, p = 0.000), and fornix (B = 10.97, p = 0.000). Lower A beta(42) levels were associated to lower FA and higher MD, AxD, and RD in the fornix, corpus callosum, inferior longitudinal fasciculus, and hippocampus. The association between AxD and A beta(42) was moderated by 7K-C (p = 0.048). Conclusion: This study adds clinical evidence to support the role of 7K-C on axonal integrity and the involvement of cholesterol metabolism in the A beta(42) generation process.

[EN] The Tear Film Lipid Layer (TFLL) acts primarily as an interface between the aqueous layer and air. Tear film lipid is composed of a thin layer of polar lipids that interact with the secretory layer of the underlying mucosa and a thicker layer of non-polar lipids at the air interface. The tear film has a complex structure and composition that protects the cornea, promotes wound healing, and maintains high-quality vision. Plasma Rich in Growth Factor (PRGF) eye drops emerged as an exciting new treatment for corneal epitheliopathies, including aqueous deficient dry eye. The purpose of this study was to compare the lipidomic profile of eye drops obtained from PRGF with tear lipidome to determine whether PRGF drops could be an adequate complement to tears in patients with impaired TFLL. To address this study, tears and blood was collected and processed from healthy donors to obtain PRGF eye drops. Samples were aliquoted and stored at -80°C until use. The lipid profiles of these samples were analysed by Ultrahigh Performance Liquid Chromatography (UHPLC) using a Vanquish UHPLC system to obtain untargeted lipidome profiles on a Q-Exactive HF-X hybrid quadrupole-Orbitrap mass spectrometer. In PRGF eye drops, 408 lipids were identified in ESI+mode and 183 in ESI- mode, and they were grouped into 15 different lipid classes from four distinct categories. By contrast, 112 lipid species were identified from tear samples in ESI+mode and 36 in ESI- mode, belonging to 12 lipid classes from six different categories. The relative abundance of most lipid species was much greater in the PRGF eye drops than in the tear, although there were some lipids present in tears that were not found in the PRGF, such as wax esters and (O-acyl)-omega-hydroxy fatty acids. In summary, these results suggest that the lipids present in PRGF eye drops could serve as a tear supplement in individuals in whom tear lipid composition is altered, although there are differences in the lipid profile of these two fluids. The authors wish to acknowledge the financial support received to carry out this work from: MINECO-Retos Fondos Fender (RTC-2016-48231), Gobierno Vasco (PUE_2018_1_0004), ELKARTEK (KK-2019/00086), PIBA 2020-1-0026, MINECO-Retos (PID2019-111139RB-I00), ELKARTEK (KK-2021-00023) to EV and FISS-21-RD21/0002/0041 to AA.

This work intends to describe the physical properties of red blood cell (RBC) membranes in obese adults. The hypothesis driving this research is that obesity, in addition to increasing the amount of body fat, will also modify the lipid composition of membranes in cells other than adipocytes. Forty-nine control volunteers (16 male, 33 female, BMI 21.8 ± 5.6 and 21.5 ± 4.2 kg/m2, respectively) and 52 obese subjects (16 male and 36 female, BMI 38.2± 11.0 and 40.7 ± 8.7 kg/m2, respectively) were examined. The two physical techniques applied were atomic force microscopy (AFM) in the force spectroscopy mode, which allows the micromechanical measurement of penetration forces, and fluorescence anisotropy of trimethylammonium diphenylhexatriene (TMA-DPH), which provides information on lipid order at the membrane polar–nonpolar interface. These techniques, in combination with lipidomic studies, revealed a decreased rigidity in the interfacial region of the RBC membranes of obese as compared to control patients, related to parallel changes in lipid composition. Lipidomic data show an increase in the cholesterol/phospholipid mole ratio and a decrease in sphingomyelin contents in obese membranes. ω-3 fatty acids (e.g., docosahexaenoic acid) appear to be less prevalent in obese patient RBCs, and this is the case for both the global fatty acid distribution and for the individual major lipids in the membrane phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS). Moreover, some ω-6 fatty acids (e.g., arachidonic acid) are increased in obese patient RBCs. The switch from ω-3 to ω-6 lipids in obese subjects could be a major factor explaining the higher interfacial fluidity in obese patient RBC membranes., This work was supported in part by the Basque Government Department of Economic Development, grant No. KK-2019/00028 (OBINTER); the Basque Government Department of Education, grants No. IT1264-19, IT1281-19, IT1270-19, and IT1625-22; the Basque Government Department of Health, grants No. 2019-222030, 2020-333023; Fundación Ramón Areces; and by Centre for the Development of Industrial Technology (CDTI) of the Spanish Ministry of Science and Innovation under the grant agreement: TECNOMIFOOD project (CER-20191010) and Basque Government: IT1625-22.

People across the world and throughout history have gone to great lengths to enhance their physical appearance. Evolutionary psychologists and ethologists have largely attempted to explain this phenomenon via mating preferences and strategies. Here, we test one of the most popular evolutionary hypotheses for beauty-enhancing behaviors, drawn from mating market and parasite stress perspectives, in a large cross-cultural sample. We also test hypotheses drawn from other influential and non-mutually exclusive theoretical frameworks, from biosocial role theory to a cultural media perspective. Survey data from 93,158 human participants across 93 countries provide evidence that behaviors such as applying makeup or using other cosmetics, hair grooming, clothing style, caring for body hygiene, and exercising or following a specific diet for the specific purpose of improving ones physical attractiveness, are universal. Indeed, 99% of participants reported spending >10 min a day performing beauty-enhancing behaviors. The results largely support evolutionary hypotheses: more time was spent enhancing beauty by women (almost 4 h a day, on average) than by men (3.6 h a day), by the youngest participants (and contrary to predictions, also the oldest), by those with a relatively more severe history of infectious diseases, and by participants currently dating compared to those in established relationships. The strongest predictor of attractiveness-enhancing behaviors was social media usage. Other predictors, in order of effect size, included adhering to traditional gender roles, residing in countries with less gender equality, considering oneself as highly attractive or, conversely, highly unattractive, TV watching time, higher socioeconomic status, right-wing political beliefs, a lower level of education, and personal individualistic attitudes. This study provides novel insight into universal beauty-enhancing behaviors by unifying evolutionary theory with several other complementary perspectives., National Science Center, Poland [2019/33/N/HS6/00054]; Basic Research Program at the National Research University Higher School of Economics (HSE University); Basic Research Program at HSE University, RF; FCT [UID/PSI/03125/2021, SFRH/BD/126304/2016]; UTAR Research Centre Excellence Award; Universiti Tunku Abdul Rahman [2019 -CAP (6401/0019)], This work is the result of the research project funded by the National Science Center, Poland (2019/33/N/HS6/00054). Dmitry Grigoryev was supported by the Basic Research Program at the National Research University Higher School of Economics (HSE University).; Dmitrii Dubrov was supported by the Basic Research Program at HSE University, RF.; Patricia Arriaga was supported by the FCT through funds from the research center UID/PSI/03125/2021. Anabela C. Santos was supported by the FCT through funds from a PhD grant SFRH/BD/126304/2016. Kavitha Nalla Muthu and Chee-Seng Tan were supported by the UTAR Research Centre Excellence Award 2019 -CAP (6401/0019) from the Universiti Tunku Abdul Rahman.

Background Information is lacking regarding long-term survival and predictive factors for mortality in patients with acute hypoxemic respiratory failure due to coronavirus disease 2019 (COVID-19) and undergoing invasive mechanical ventilation. We aimed to estimate 180-day mortality of patients with COVID-19 requiring invasive ventilation, and to develop a predictive model for long-term mortality. Methods Retrospective, multicentre, national cohort study between March 8 and April 30, 2020 in 16 intensive care units (ICU) in Spain. Participants were consecutive adults who received invasive mechanical ventilation for COVID-19. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection detected in positive testing of a nasopharyngeal sample and confirmed by real time reverse-transcriptase polymerase chain reaction (rt-PCR). The primary outcomes was 180-day survival after hospital admission. Secondary outcomes were length of ICU and hospital stay, and ICU and in-hospital mortality. A predictive model was developed to estimate the probability of 180-day mortality. Results 868 patients were included (median age, 64 years [interquartile range [IQR], 56–71 years]; 72% male). Severity at ICU admission, estimated by SAPS3, was 56 points [IQR 50–63]. Prior to intubation, 26% received some type of noninvasive respiratory support. The unadjusted overall 180-day survival rates was 59% (95% CI 56–62%). The predictive factors measured during ICU stay, and associated with 180-day mortality were: age [Odds Ratio [OR] per 1-year increase 1.051, 95% CI 1.033–1.068)), SAPS3 (OR per 1-point increase 1.027, 95% CI 1.011–1.044), diabetes (OR 1.546, 95% CI 1.085–2.204), neutrophils to lymphocytes ratio (OR per 1-unit increase 1.008, 95% CI 1.001–1.016), failed attempt of noninvasive positive pressure ventilation prior to orotracheal intubation (OR 1.878 (95% CI 1.124–3.140), use of selective digestive decontamination strategy during ICU stay (OR 0.590 (95% CI 0.358–0.972) and administration of low dosage of corticosteroids (methylprednisolone 1 mg/kg) (OR 2.042 (95% CI 1.205–3.460). Conclusion The long-term survival of mechanically ventilated patients with severe COVID-19 reaches more than 50% and may help to provide individualized risk stratification and potential treatments. Trial registration: ClinicalTrials.gov Identifier: NCT04379258. Registered 10 April 2020 (retrospectively registered)

The cerebrospinal fluid (CSF) lipidome is attracting increasing attention due to the importance of lipids in brain molecular signaling and their involvement in several neurological diseases. Different solvent systems have been used for the extraction of multiple lipid classes from CSF but no comparative study of the effectiveness of these protocols has been carried out. To optimize CSF lipid extraction for lipidomic measurements by untargeted ultra-high performance liquid chromatography - mass spectrometry, we evaluate and compare two sample preparation protocols, one involving protein precipitation with isopropanol (IPA) and other consisting of a liquid-liquid extraction with chloroform-methanol. For that purpose, human CSF from neurologically healthy and normolipidemic volunteers was used. The criteria established to compare these two methods were based on four critical aspects of sample preparation: simplicity, lipid coverage, reproducibility and recovery efficiencies. We found that both methods were highly reproducible techniques (>75% of the lipids with coefficient of variation (CV)

Background COVID-19 patients can develop a cytokine release syndrome that eventually leads to acute respiratory distress syndrome (ARDS) requiring invasive mechanical ventilation (IMV). Since interleukin-6 (IL-6) is a relevant cytokine in ARDS, the blockade of its receptor with Tocilizumab (TCZ) could reduce mortality and/or morbidity in severe COVID-19. Objective To determine whether baseline IL-6 serum levels can predict the need for IMV and the response to TCZ. Methods Retrospective observational study performed in hospitalized patients diagnosed of COVID-19. Clinical information and laboratory findings, including IL-6 levels, were collected approximately 3 and 9 days after admission to be matched with pre- and post-administration of TCZ. Multivariable logistic and linear regressions, and survival analysis were performed depending on outcomes: need for IMV, evolution of arterial oxygen tension/fraction of inspired oxygen ratio (PaO2/FiO2) or mortality. Results One hundred and forty-six patients were studied, predominantly male (66%); median age was 63 years. Forty-four patients (30%) required IMV, and 58 patients (40%) received treatment with TCZ. IL-6 levels>30 pg/ml was the best predictor for IMV (OR:7.1; p30 pg/ml predicts IMV requirement in patients with COVID-19 and contributes to establish an adequate indication for TCZ administration., Baseline IL-6 serum levels>30 pg/ml identify severe COVID-19 patients and should be used to guide the intervention with IL-6R inhibitors, aiming to improve their use in an uncertain and evolving therapeutic scenario.

Here we present a simple and cost effective procedure to improve the spatial resolution of the commercial MALDI source of a LTQ Orbitrap. Based in spatial filtering techniques, we demonstrate that, with minimal modifications of the original set up, the system resolution can be pushed forward to10 μm. The improved system performance is demonstrated by means of MALDI imaging of human colon biopsies.

Melanoma is the deadliest form of skin cancer due to its ability to colonize distant sites and initiate metastasis. Although these processes largely depend on the lipid-based cell membrane scaffold, our understanding of the melanoma lipid phenotype lags behind most other aspects of this tumor cell. Here, we examined a panel of normal human epidermal and nevus melanocytes and primary and metastatic melanoma cell lines to determine whether distinctive cell-intrinsic lipidomes can discern non-neoplastic from neoplastic melanocytes and define their metastatic potential. Lipidome profiles were obtained by UHPLC-ESI mass-spectrometry, and differences in the signatures were analyzed by multivariate statistical analyses. Significant and highly specific changes in more than 30 lipid species were annotated in the initiation of melanoma, whereas less numerous changes were associated with melanoma progression and the non-malignant transformation of nevus melanocytes. Notably, the “malignancy lipid signature” features marked drops in pivotal membrane lipids, like sphingomyelins, and aberrant elevation of ether-type lipids and phosphatidylglycerol and phosphatidylinositol variants, suggesting a previously undefined remodeling of sphingolipid and glycerophospholipid metabolism. Besides broadening the molecular definition of this neoplasm, the different lipid profiles identified may help improve the clinical diagnosis/prognosis and facilitate therapeutic interventions for cutaneous melanoma. This research was funded in part by grants from the Ministry of Economy; Industry and Competitiveness (RTC-2015-3693-1); Ministry of Science and Innovation (RTI-2018-095134-B-I00); Basque Government (IT971-16; IT1162-19; KK2016-036; KK2017-041 and KK2018-00090) and UPV/EHU (GIU17/066).

The data contain information related to the research article entitled "Profiling of promoter occupancy by the SND1 transcriptional coactivator identifies downstream glycerolipid metabolic genes involved in TNF alpha response in human hepatoma cells" (DOI: 10.1093/nar/gkv858). In the article alluded to, we reported that tumor necrosis factor alpha (TNF alpha) increases notably the cellular content of the major glycerolipid phosphatidylcholine (PC). Here, accompanying lipidomic data determine the PC structural variants that have been identified in human hepatoma HepG2 cells and those whose relative abundance is modified by TNF alpha. We used ultrahigh performance liquid chromatography (UHPLC) coupled to electrospray ionization (ESI) tandem mass spectrometry (MS/MS)-based lipidomic profiling to analyze lipid extracts of control and TNF alpha-treated HepG2 cells. The identity of PC individual species was elucidated using the values of the retention time and molecular weight in addition to the fragmentation patterns. MS data were then processed and analyzed for the characterization of statistically significant differences in detected structural variants. We have annotated the dataset of PC species that characterize HepG2 cells' phenotype, both under normal and pro-inflammatory conditions. This work was supported by the Basque Government Departments of Education (grant IT971-16) and Economic Promotion (grant KK2018/00090), Basque Country, Spain.

In this paper we describe how incorporating inter-well flow of gas tracers into a numerical simulation model of an oil producing field allowed us to improve the reservoir characterization. Gas tracers evidenced the lateral and vertical reservoir connectivity, identified preferential flow paths and eventually provided an additional tool for the dynamic history match. The field produces light oil with the support of miscible gas injection, from a reservoir composed of two stacked fluvial sandstones units. To improve the reservoir characterization four inter-well gas tracer campaigns involving a total of 13 injector wells have been successfully completed. All these tracer injections have then been modeled at our full field numerical simulation model with the purpose of challenging the reservoir description in it. Input from this exercise have been later-on used during the construction of geomodels, benefiting from our improved reservoir knowledge. The reservoir is composed of two units, Middle (M) and Upper (U), both deposited as laterally amalgamated fluvial channels. Both units appear as vertically separated by an impervious shale interval, only absent in 2 wells out of 30. The shale interval was originally considered as a possible flow barrier by the earliest geological models. However, numerical simulation models were only able to replicate observed tracers arrivals when specific vertical connections existed between both units, indicating the shale interval was no as laterally continuous as formerly suspected. History matching the tracers arrivals in the western field area was also helpful to reveal a fast gas breakthrough between wells which were aligned perpendicularly to the main channel orientation (NW-SE). This finding confirmed that this prevailing channel orientation was not the only responsible for a good reservoir communication, but also the lateral amalgamation (SW-NE) of channels was exerting a significant control in low sinuosity fluvial systems, as well as secondary flow directions in high sinuosity systems. The improved reservoir characterization have been reflected in subsequent reservoir geological models and numerical simulators, avoiding misleading history matching solutions. Also, it is worth to note that this have had a direct impact on the gas injection strategy followed in the field. Quite commonly interwell tracer flow is not fully incorporated into numerical simulators. This is a time consuming process, which, in addition to the conventional model uncertainties, requires sensitivities to the associated tracer parameters. This paper demonstrate through a real case how valuable this additional effort may result, and how it may improve the geological and dynamic understanding of our field.

Lipids are essential components of cells and tissues. They play active and central roles in signaling and many biological functions and therefore their dysregulation is very often the first signal of function alteration. Here we describe the protocol to analyze not only lipid expression in rat sciatic nerve but also the lipid distribution along its different anatomic areas. The protocol combines results from MALDI-IMS and UHPLC-MS/MS to identify and cartography the maximum number of lipid species in the tissue.

In the present work, a comparative study of the detailed phenolic profile of fruit and juice samples from an Asturian cider apple breeding program was carried out. To achieve this, a fast, selective and sensitive UHPLC-DAD strategy was developed and validated. The proposed method allowed determining 49 polyphenols in apple samples and 41 phenolic compounds in apple juice samples in 18 min. Furthermore, six minor flavonoids have been quantified for the first time in apples: phloretin-hexosyl-hexoside, isorhamnetin-3-O-galactoside, isorhamnetin-3-O-glucoside, isorhamnetin-3-O-arabinopyranoside, isorhamnetin-3-O-arabinofuranoside and isorhamnetin-3-O-rhamnoside. The selectivity and reliability of this methodology allowed us to apply it in a successful manner to select the most interesting descendants from a progeny of ‘Meana’ × ‘Florina’ (a cider and a dessert apple, respectively) in terms of their phenolic composition. Descendants X9701-349, 119, 370, 120 and 069 could be proposed for cider production, since they exhibited a higher phenolic content than ‘Meana’ apple juice. Although flavan-3-ol, procyanidin and dihydrochalcone levels were similar to those observed in ‘Meana’ apple, flavonol levels increased slightly. Furthermore, it is worth noting that hydroxycinnamic acid concentration was up to three times higher. This fact could lead to an increase in cider astringency and color, as well as in cider aroma. Thus, this methodology is proposed to be applicable to the determination of the phenolic profile in other fruit and/or plant extracts.

Lipids are essential components of cells and tissues. They play active and central roles in signaling and many biological functions and therefore their dysregulation is very often the first signal of function alteration. Here we describe the protocol to analyze not only lipid expression in rat sciatic nerve but also the lipid distribution along its different anatomic areas. The protocol combines results from MALDI-IMS and UHPLC-MS/MS to identify and cartography the maximum number of lipid species in the tissue.

Human tumor xenografts in immunodeficient mice are a very popular model to study the development of cancer and to test new drug candidates. Among the parameters analyzed are the variations in the lipid composition, as they are good indicators of changes in the cellular metabolism. Here, we present a study on the distribution of lipids in xenografts of NCI-H1975 human lung cancer cells, using MALDI imaging mass spectrometry and UHPLC-ESI-QTOF. The identification of lipids directly from the tissue by MALDI was aided by the comparison with identification using ESI ionization in lipid extracts from the same xenografts. Lipids belonging to PCs, PIs, SMs, DAG, TAG, PS, PA, and PG classes were identified and their distribution over the xenograft was determined. Three areas were identified in the xenograft, corresponding to cells in different metabolic stages and to a layer of adipose tissue that covers the xenograft.

Anthocyanins, responsible for wine colour, are involved in many reactions during wine ageing. Anthocyanin-flavanol associations give rise to derivatives in flavylium form that provide blue hues, but also derivatives that do not directly influence wine colour. These colourless derivatives remain mostly unknown but their roles during wine ageing are important for controlling wine quality. Colourless anthocyanin-flavanol derivatives formed during wine ageing have been studied in three aged red wines from Rioja using a combined method with Column Chromatography (CC) and High Performance Liquid Chromatography with Diode Array and Mass Spectrometric detections (HPLC-DADMS). Twenty-six compounds have been detected: 17 dimers with the anthocyanin in flavene form with possible anthocyanin-flavanol (type 1) and flavanol-anthocyanin (type 2) structures, and 9 with an A-type bicyclic anthocyanin-flavanol structure (type 3). Although some of malvidin derivatives have been previously reported, this is the first time that these derivatives (including different isomers) have also been detected for delphinidin, petunidin and peonidin.

The contents of 49 polyphenols in sweet orange, tangerine, lemon and grapefruit juices from 18 cultivars grown in Spain were determined by reversed-phase high-performance liquid chromatography with photodiode array detection. Citrus polyphenolic profiles consist of 81–97 % of flavanones, 0.3–13.6 % of flavones, 0.1–6.0 % of flavonols, 0.6–9.6 % of hydroxycinnamic acids and 0.2–0.4 % of coumarins (only found in grapefruit juices). Several markers that allow to distinguish with practical certainty grapefruit and lemon juices between them and from the other Citrus species are reported. Each of these markers is a reliable and useful tool to detect juice adulteration. Grapefruit juice markers were naringenin-7-O-neohesperidoside, naringenin-7-O-neohesperidoside-4′-O-glucose, naringenin-O-hexosylhexoside, hesperetin-7-O-neohesperidoside, naringenin-O-rhamnosylmalonylhexoside, isosakuranetin-7-O-neohesperidoside, hesperetin-7-O-rutinoside, apigenin-6-C-hexoside-O-hexoside, apigenin-7-O-neohesperidoside and scopoletin-O-hexoside. Lemon juice markers were eriodictyol-7-O-rutinoside-4′-O-glucoside, eriodictyol-7-O-rutinoside, diosmetin-6,8-di-C-glucoside, diosmetin-8-C-glucoside, luteolin-7-O-rutinoside, diosmetin-6-C-glucoside and diosmetin-6,8-di-C-hexosideacylhexoside. The markers naringenin-O-hexosylhexoside, apigenin-6-C-hexoside-O-hexoside, scopoletin-O-hexoside, diosmetin-8-C-glucoside and diosmetin-6,8-di-C-hexosideacylhexoside were detected, characterized and quantitatively determined in grapefruit and lemon juices for the first time by our research group, as far as the authors know. Classification models provided by LDA and PLS-DA correctly identify all sweet orange and tangerine juices. Moreover, PLS regression model determines the percentage (10–70 %) of tangerine juice used to adulterate sweet orange juice with a suitable confidence interval (RMSEP = 7 %).

A new, rapid, selective and sensitive ultrahigh performance liquid chromatography with diode array detection coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry (UHPLC-DAD–ESI-Q-ToF-MS) strategy using automatic and simultaneous acquisition of exact mass at high and low collision energy, MSE, has been developed to obtain polyphenolic profile of apples, apple pomace and apple juice from Asturian cider apples in a single run injection of 22 min. MSE spectral data acquisition overcomes chromatographic co-elution problems, performing simultaneous collection of precursor ions as well as other ions produced as a result of their fragmentation, which allows resolving complex spectra from mixtures of precursor ions in an unsupervised way and eases their interpretation. Using this technique, 52 phenolic compounds of five different classes were readily characterized in these apple extracts in both positive and negative ionization modes. The spectral data for phenolic compounds obtained using this acquisition mode are comparable to those obtained by conventional LC–MS/MS as exemplified in this work. Among the 52 phenolic compounds identified in this work, 2 dihydrochalcones and 3 flavonols have been tentatively identified for the first time in apple products. Moreover, 2 flavanols, 4 dihydrochalcones, 9 hydroxycinnamic acids and 4 flavonols had not been previously reported in apple by ToF analysis to our knowledge.

A chemometric method for the treatment of ion mobility coupled to mass spectrometry (IMS/MS) data is proposed as a complementary tool for obtaining experimental evidence for the study of MS fragmentations, which can provide a direct and automatable methodology for characterising ionic series and the hierarchy of all product ions of an MS spectrum. Two MS/MS with ion mobility experiments have been designed: in the first, the intrinsic mobility of each ion is estimated, and in the second experiment, distributions of the ionic intensity of product ions fragmented after IMS separations are recorded. These mobilograms are aligned using the coshift algorithm and mathematically fitted using Classical Least Squares (CLS) to determine the mobility contributions from their precursor ions. Despite some limitations when studying low intensity ions and ions with similar ion mobility, CLS fitting improves the usage of IMS coupled with accurate mass spectrometry as a complementary tool in the study of MS fragmentation mechanisms and more notably, it offers an automatable and efficient alternative to MS3 experiments.

A mass spectrometric method using electrospray ionization with triple quadrupole and quadrupole time-of-flight hybrid (Q-Tof) mass spectrometry has been applied to the structural characterization of dihydroflavonols. This family of compounds has been studied by liquid chromatography/tandem mass spectrometry (LC/MS/MS) for the first time in this work. A comprehensive study of the product ion MS spectra of the [M+H](+) ion of a commercially available standard has been performed. The most useful fragmentations in terms of structural identification are those that involve cleavage of the C-ring, resulting in diagnostic ions of dihydroflavonol family: (1,3)A(0) (+), (1,2)B(0) (+), (1,2)B(0) (+)-CO, (0,2)A(0) (+), (0,2)A(0) (+)-H(2)O, (0,2)A(0) (+)-CO, and (0,2)A(0) (+)-H(2)O-CO, that allow the characterization of the substituents in the A- and B-rings. In addition to those ions, other product ions due to losses of H(2)O and CO molecules from the Y(0) (+) ion were observed. Their fragmentation mechanisms and ion structures have been proposed. The established fragmentation patterns have been used to successfully identity three dihydroflavonols found in tangerine juices for the first time.

In the present study, a methodology based on liquid chromatography with diode array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer for the simultaneous identification of phenolic compounds in fruit juices has been developed. 72 available phenolic compound standards from diverse families present in fruits have been studied in order to analyze their fragmentation pattern. As a result, a general strategy for the characterization of unknown phenolic compounds in fruit juices was designed: (i) taking into account its UV-visible spectrum and elution order, assign the unknown polyphenol to a polyphenol class, (ii) identify the quasi-molecular ion using positive and negative MS spectra, being supported by adducts generated with solvent or sodium and molecular complexes, (iii) determinate the pattern of glycosylation in positive mode using ESI(+)-CID MS/MS product ion scan experiments, selecting the quasi-molecular ion as precursor ion, and finally, (iv) study the identity of the aglycone through ESI(+)-CID MS/MS product ion spectra from the protonated aglycone, [Y(0)](+). This strategy was successfully employed for the characterization of known and unknown phenolic compounds in juices from 17 different fruits.

Fifteen flavonoid O-diglycosides with different interglycosidic linkage isomery and glycosylation position have been studied in order to analyze their fragmentation patterns. Initial separation was carried out using high performance liquid chromatography with diode array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer. Some useful differences in their MS spectra have been found and discussed. As it has already been reported, [Y*]+/[Y0]+ ratio for flavanones and [Y1]+/[Y0]+ ratio for other flavonoids is specific for each isomeric interglycosidic linkage. In this work it has also been observed that the abundance of these ions is dependent on the position of glycosylation. On the basis of these differences, systematic guidelines for our experimental conditions have been proposed for the differentiation of not only isomeric interglycosidic linkage but also glycosylation position using collision-induced dissociation MS/MS (CID-MS/MS) spectra in positive mode. These results have been successfully applied for the characterization of three diglycosyl flavonoids found in Citrus fruit juices and these conclusions have also been extrapolated for characterizing two triglycosides in the same fruits.

Six flavone mono-C-glucosides, four standards (beta-D-glucopyranosyl-(1 --> C-6)- and -(1 --> C-8)- apigenin and luteolin) and two others from lemon juice (beta-D-glucopyranosyl-(1 --> C-6)- and -(1 --> C-8)-diosmetin) have been studied in order to analyze their fragmentation patterns. Initial separation was carried out using high-performance liquid chromatography with diode-array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer. Several systematic differences between collision-induced dissociation tandem mass (CID-MS/MS) spectra of C-6- and C-8-isomers have been found and some general guidelines and two new diagnostic product ions have been proposed for the differentiation of C-6- and C-8-flavonoid glycosides. These results have been successfully applied to the characterization of two flavone C-glycosides found in lemon juice, and mass spectra of a flavone di-C-glycoside detected in lemon juice have been studied and interpreted.

OBJETIVOS: Identificar las principales características sociodemográficas, clínicas y conductuales de los pacientes que participaban en los programas de atención al diabético en Medellín, Colombia, y evaluar el cumplimiento de las metas de control metabólico y de los estándares de control en el diabético. MÉTODOS: Se realizó un estudio descriptivo transversal mediante el análisis de las historias clínicas de todos los pacientes que asistían a los programas de atención al diabético desde al menos 6 meses en nueve instituciones de salud entre enero de 2001 y diciembre de 2003. Se tomaron datos sociodemográficos (edad y sexo), clínicos (tiempo de evolución de la enfermedad, tipo y tratamiento de la diabetes y enfermedades concurrentes y su tratamiento), sobre los hábitos de vida (práctica de ejercicios físicos, hábito de fumar y consumo de azúcar y grasas) y sobre las complicaciones crónicas y los resultados de las pruebas de laboratorio del último año. Todos los datos confusos o faltantes se registraron como ausentes. RESULTADOS: De las 3 583 historias clínicas evaluadas, se logró conocer el tipo de diabetes que padecían 3 554 pacientes, de ellos 95,1% correspondieron a pacientes con diabetes tipo 2. En general, 56,9% (intervalo de confianza de 95% [IC95%]: 55,2 a 58,6%) de los pacientes realizaban ejercicios periódicos; 15,1% (IC95%: 13,9 a 16,3%) aún fumaba en el momento del corte, 17,7% (IC95%: 16,4 a 19,0%) había dejado de fumar y 67,2% (IC95%: 65,6 a 68,8%) nunca había fumado. En total, 21,0% (IC95%: 19,6 a 22,5%) de los pacientes consumía azúcar regularmente, mientras 24,8% (IC95%: 23,3 a 26,3%) consumía grasas y solo 19,5% (IC95%: 17,8 a 21,3%) realizaba automonitoreo de la glucemia con una frecuencia semanal o mayor. La prevalencia general de hipertensión arterial fue de 68,8% y de 98,2% de dislipidemia en los diabéticos tipo 2. El promedio general del índice de masa corporal fue de 28,0 kg/m² (IC95%: 27,8 a 28,2). El promedio del número de medicamentos utilizados fue de 1,3 para el tratamiento de la diabetes tipo 2, de 1,9 para la hipertensión arterial y de 0,6 para la dislipidemia. No se encontraron los datos de dislipidemia de colesterol de lipoproteínas de baja densidad (LDL) en 44,8% de las historias clínicas y de colesterol de lipoproteínas de alta densidad (HDL) en 16,4%. Las complicaciones oculares fueron las más frecuentes (31,8%; IC95%: 30,1 a 33,5%), seguidas de las afecciones renales (25,9%; IC95%: 24,4 a 27,5%) y las cardiovasculares (22,5%; IC95%: 21,1 a 23,8%). El cumplimiento de las metas de control del colesterol de LDL, los triglicéridos y el colesterol de HDL fue insuficiente (14,2%, 36,9% y 47,4%, respectivamente). CONCLUSIONES: En la población estudiada, las tres complicaciones crónicas que más influyen en el pronóstico y el costo de salud en los diabéticos fueron la nefropatía, la retinopatía y la dislipidemia. El gran subregistro de datos encontrado en las historias clínicas se puede reducir si se toman medidas para estandarizar las historias clínicas y se realizan evaluaciones periódicas de la calidad de las mismas.OBJECTIVES: To identify the principal sociodemographic, clinical, and behavioral characteristics of patients participating in diabetic care programs in the city of Medellín, Colombia, and to evaluate progress toward the goal of metabolic control and diabetic care standards. METHODS: A cross-sectional, descriptive study was done, analyzing the clinical records of patients who had been participating for at least 6 months in the diabetic care programs of nine health care centers in Medellín. The study period ran from January 2001 to December 2003. The following data were collected: sociodemographic (age and sex), clinical (time since disease onset, diabetes type and treatment, and concurrent illnesses and their treatment), lifestyle habits (exercise routine, smoking, and sugar and fat consumption), chronic complications, and laboratory test results from the prior year. Questionable or missing data were categorized as unavailable. RESULTS: Of the 3 583 clinical histories evaluated, we were able to confirm the form of diabetes for 3 554 patients. Of those 3 554, 95.1% had type 2 diabetes. Overall, 56.9% (95% confidence interval (95% CI): 55.2% to 58.6%) of the patients exercised regularly, 15.1% (95% CI: 13.9% to 16.3%) were current smokers, 17.7% (95% CI: 16.4% to 19.0%) were former smokers, and 67.2% (95% CI: 65.6% to 68.8%) had never smoked. In all, 21.0% (95% CI: 19.6% to 22.5%) of the patients consumed sugar regularly, while 24.8% (95% CI: 23.3% to 26.3%) consumed fats and only 19.5% (95% CI: 17.8% to 21.3%) were self-monitoring their glucose levels at least weekly. Among the type 2 diabetics, 68.8% had high blood pressure, and 98.2% had dyslipidemia. The average body mass index was 28.0 kg/m² (95% CI: 27.8 to 28.2). The average number of medications that the patients were taking to treat type 2 diabetes was 1.3; to treat high blood pressure, 1.9; and to treat dyslipidemia, 0.6. There were no data on low-density lipoprotein (LDL) cholesterol in 44.8% of the clinical records, and no data on high-density lipoprotein (HDL) cholesterol in 16.4% of the records. Among the complications, the most common were ocular ones (31.8%; 95% CI: 30.1% to 33.5%), followed by renal ones (25.9%; 95% CI: 24.4% to 27.5%), and cardiovascular ones (22.5%; 95% CI: 21.1% to 23.8%). Cholesterol control achievement was inadequate for LDL (reached by only 14.2% of the patients), for triglycerides (36.9% of patients), and HDL (47.4% of patients). CONCLUSIONS: Among the study population in Medellín, the three chronic complications with the greatest impact on prognosis and health care costs of the diabetics were nephropathy, retinopathy, and dyslipidemia. The noticeable amount of unavailable data in the clinical records could be greatly reduced by standardizing the clinical record forms and by periodic quality checks of the records themselves.

Sets of hetero- and homoannularly bridged ferrocenyl aminophosphine and diphosphine ligands were investigated in palladium-catalysed allylic alkylation and amination reactions ([1-PPh2-2,1′-(α-R-CH(CH2)2)-ferrocene]: (Rc,Rp)-1: R = N(CH3)2; (Rc,Rp)-4: R = PPh2; (Rc,Rp)-5: R = PCy2; ([1-PPh2-2,3-(α-R-CH(CH2)3)-ferrocene]: (Rc,Rp)-2: R = N(CH3)2; (Sc,Rp)-3: R = N(CH3)2; (Sc,Rp)-6: R = PPh2; (Sc,Rp)-7: R = PCy2). Diphenylprop-2-en-1-yl acetate and pent-3-en-2-yl acetate were used as the substrates and dimethyl malonate or benzylamine as the nucleophiles. Catalytic data were analysed and compared to those of PPFA (8) and Josiphos-type (9, 10) ligands ([1-PPh2-2-(α-R-CHCH3)-ferrocene]: (Sc,Rp)-8: R = N(CH3)2; (Sc,Rp)-9: R = PPh2; (Sc,Rp)-10: R = PCy2). Correlations between changes in enantioselectivity or absolute configuration of product and changes in ligand backbones or functional groups were assessed. Cationic palladium(II) diphenylallyl complexes of ligands 1, 4, 5 and 7 were isolated and their conformational behaviour in solution was analysed both as the isolated complexes and under catalytic conditions. The molecular structure of complex 7C (endo syn/syn form) was determined by X-ray diffraction.

A series of xylodextrins has been produced by enzymatic or hydrothermal degradation of industrial xylans. For further synthetic use, the oligomers were converted into per-O-acetylated xylooligomers which were separated by silica gel chromatography to furnish preparative amounts of xylobiose up to xylopentaose. In a model reaction, selective anomeric deacetylation and treatment with trichloroacetonitrile furnished a xylobiosyl donor, which was converted into the β-methyl glycoside. In addition, methyl β-D-xylopyranoside was transformed into a suitable glycosyl acceptor via tosylation followed by a double displacement reaction at 0-4, allowing for further chain elongation and modification at the reducing xylopyranosyl unit.

The polyphenolic compositions of 31 Basque cider apple cultivars were determined in pulp, peel, and juice by high-performance liquid chromatography with diode array detection analysis of crude extracts and after thiolysis. Total polyphenols are distributed in a wide concentration range depending on the cultivar. Procyanidins are the class of polyphenols that present major concentrations in apple. Their average degrees of polymerization range from 4 to 8 depending on the cultivar. Apple cultivars were technologically classified into bitter and nonbitter categories using different classification systems obtained by applying several pattern recognition techniques, such as principal component analysis, K-nearest neighbors, soft independent modeling of class analogy, partial least-squares, and multilayer feed-forward-artificial neural networks, to apple pulp, peel, or juice data (individual polyphenol concentrations, total procyanidin content, and the average degree of polymerization of procyanidins). Bitter apple cultivars present higher contents of flavan-3-ols and/or dihydrochalcones than nonbitter cultivars. Detailed knowledge of the polyphenolic profile of each apple cultivar affords information about their susceptibility to oxidation, their sensory properties (bitterness, astringency), and their possible influence on the characteristics and quality of the final product (juice, cider) when apples are processed.

The data set composed by phenolic compound profiles of 83 Citrus juices (determined by HPLC-DAD-MS/MS) was evaluated by chemometrics to differentiate them according to Citrus species (sweet orange, tangerine, lemon, and grapefruit). Cluster analysis (CA) and principal component analysis (PCA) showed natural sample grouping among Citrus species and even the Citrus subclass. Most of the information contained in the full data set can be captured if only 15 phenolic compounds (concentration ≥10 mg/L), which can be quantified with fast and accurate methods in real samples, are introduced in the models; a good classification which allows the confirmation of the authenticity of juices is achieved by linear discriminant analysis. Using this reduced data set, fast and routine methods have been developed for predicting the percentage of grapefruit in adulterated sweet orange juices using principal component regression (PCR) and partial least-squares regression (PLS). The PLS model has provided suitable estimation errors.