83 research outputs found
Fabric phase sorptive extraction followed by ultra-performance liquid chromatography-tandem mass spectrometry for the determination of fungicides and insecticides in wine
This is the Author’s Accepted Manuscript of the following article: Pérez-Mayán, L., Rodríguez, I., Ramil, M., Kabir, A., Furton, K., & Cela, R. (2019). Fabric phase sorptive extraction followed by ultra-performance liquid chromatography-tandem mass spectrometry for the determination of fungicides and insecticides in wine. Journal Of Chromatography A, 1584, 13-23. doi: 10.1016/j.chroma.2018.11.025In this work, fabric phase sorptive extraction (FPSE) is investigated for the extraction and preconcentration of ultra-trace level residues of fungicides (19 compounds) and insecticides (3 species) in wine samples. Subsequently, the preconcentrated analytes are selectively determined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Parameters affecting the efficiency and repeatability of the extraction are evaluated in depth; moreover, the proposed method is characterized in terms of linear response range, trueness, precision and limits of quantification (LOQs). The set-up of the extraction process and the type of coating were the variables exerting the most prominent effects in the repeatability and the yield of the extraction, respectively. Under optimized conditions, samples (10 mL of wine diluted with the same volume of ultrapure water) were extracted with a small amount of cellulose fabric (3 discs with 4 mm of diameter: total surface 0.38 cm2) coated with a sol-gel polyethylene glycol sorbent (sorbent amount 3.3 mg), immersed in the diluted sample, without being in direct contact with the PTFE covered magnetic stir bar. Following the overnight extraction step, analytes were quantitatively recovered using only 0.3 mL of an ACN-MeOH (80:20) mixture. Under equilibrium sampling conditions, the linear response range of the method varied from 0.2 to 200 ng mL−1, with limits of quantification (LOQs) between 0.03 and 0.3 ng mL−1. Relative recoveries ranged from 77 ± 6% to 118 ± 4%, and from 87 ± 4% to 121 ± 6% for red and white wines, respectively. Application of the optimized method to commercial wines demonstrated the existence of up to 9 out of 22 investigated compounds in the same wine sample. The compound identified at the highest concentration was iprovalicarb (IPR), with a value of 130 ± 9 ng mL−1 in a commercial white wineThis study has been supported by the Spanish Government, Xunta de Galicia and E.U. FEDER funds (projects CTQ2015-68660-P and GRC-ED431C). L.P-M acknowledges a FPU contract to the Spanish Ministry of EducationS
Molecular and cytogenetic characterization of expanded B-cell clones from multiclonal versus monoclonal B-cell chronic lymphoproliferative disorders
This is an open-access paper.-- et al.Chronic antigen-stimulation has been recurrently involved in the earlier stages of monoclonal B-cell lymphocytosis, chronic lymphocytic leukemia and other B-cell chronic lymphoproliferative disorders. The expansion of two or more B-cell clones has frequently been reported in individuals with these conditions; potentially, such coexisting clones have a greater probability of interaction with common immunological determinants. Here, we analyzed the B-cell receptor repertoire and molecular profile, as well as the phenotypic, cytogenetic and hematologic features, of 228 chronic lymphocytic leukemia-like and non-chronic lymphocytic leukemia-like clones comparing multiclonal (n=85 clones from 41 cases) versus monoclonal (n=143 clones) monoclonal B-cell lymphocytosis, chronic lymphocytic leukemia and other B-cell chronic lymphoproliferative disorders. The B-cell receptor of B-cell clones from multiclonal cases showed a slightly higher degree of HCDR3 homology than B-cell clones from mono clonal cases, in association with unique hematologic (e.g. lower B-lymphocyte counts) and cytogenetic (e.g. lower frequency of cytogenetically altered clones) features usually related to earlier stages of the disease. Moreover, a subgroup of coexisting B-cell clones from individual multiclonal cases which were found to be phylogenetically related showed unique molecular and cytogenetic features: they more frequently shared IGHV3 gene usage, shorter HCDR3 sequences with a greater proportion of IGHV mutations and del(13q14.3), than other unrelated B-cell clones. These results would support the antigen-driven nature of such multiclonal B-cell expansions, with potential involvement of multiple antigens/epitopes.AH was supported by a grant from the Fundação Para a Ciência e Tecnologia of Portugal (SFRH/BD/31609/2006); ARC was partly supported by a grant from Fundación Científica de la Asociación Española contra el Cáncer (AECC-2008) and by a grant from Red Temática de Investigación Cooperativa en Cáncer del Instituto de Salud Carlos III - FEDER (RD12/0036/0048). The research was supported by the following grants: Red Temática de Investigación Cooperativa en Cáncer (RTICC) del Instituto de Salud Carlos III - FONDOS FEDER (RD06/0020/0035 and RD12/0036/0048); FIS PI06/0824-FEDER, PS09/02430-FEDER and FIS PI12/00905-FEDER, from the Fondo de Investigación
Sanitaria, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain; GRS206/A/08 from the Gerencia Regional de Salud de Castilla y León and Ayuda al Grupo GR37 de Excelencia de Castilla y León, Consejería de Educación; SAN/1778/2009, Consejería de Sanidad, Junta de Castilla y León, Valladolid, Spain and FS/1-2010 Fundación Memoria D. Samuel Solórzano, Universidad de Salamanca, Salamanca, Spain.Peer Reviewe
A systematic approach for peptide characterization of B-cell receptor in chronic lymphocytic leukemia cells
A wide variety of immunoglobulins (Ig) is produced by the immune system thanks to different mechanisms (V(D)J recombination, somatic hypermutation, and antigen selection). The profiling of Ig sequences (at both DNA and peptide levels) are of great relevance to developing targeted vaccines or treatments for specific diseases or infections. Thus, genomics and proteomics techniques (such as Next- Generation Sequencing (NGS) and mass spectrometry (MS)) have notably increased the knowledge in Ig sequencing and serum Ig peptide profiling in a high-throughput manner. However, the peptide characterization of membrane-bound Ig (e.g., B-cell receptors, BCR) is still a challenge mainly due to the poor recovery of mentioned Ig. Herein, we have evaluated three different sample processing methods for peptide sequencing of BCR belonging to chronic lymphocytic leukemia (CLL) B cells identifying up to 426 different peptide sequences (MS/MS data are available via ProteomeXchange with identifier PXD004466). Moreover, as a consequence of the results here obtained, recommended guidelines have been described for BCR-sequencing of B-CLL samples by MS approaches. For this purpose, an in-house algorithm has been designed and developed to compare the MS/MS results with those obtained by molecular biology in order to integrate both proteomics and genomics results and establish the steps to follow when sequencing membrane-bound Ig by MS/MS.We gratefully acknowledge financial support from the Spanish Health Institute Carlos III (ISCIII) for the grants: FIS PI11/02114 and FIS PI114/01538. We also acknowledge Fondos FEDER (EU) and Junta Castilla León (grant BIO/SA07/15). This work has been also sponsored by Fundación Solórzano (FS/23-2015). The Proteomics Unit belongs to ProteoRed, PRB2-ISCIII, supported by grant PT13/0001, of the PE I+D+I 2013-2016, funded by ISCIII and FEDER. The authors would like to thank all the clinicians and technicians in the Cytometry and Cell Purification Services of the University of Salamanca, the Spanish National DNA Bank (Banco Nacional de DNA Carlos III, University of Salamanca) and the Genomic Unit of Cancer Research Centre (IBMCC,
USAL-CSIC) for their support in the data collection for the preparation of this manuscript. P.D. is supported by a JCYL-EDU/346/2013 Ph.D. scholarship.Peer Reviewe
Subjects with chronic lymphocytic leukaemia-like B-cell clones with stereotyped B-cell receptors frequently show MDS-associated phenotypes on myeloid cells
An increasing body of evidence suggests the potential occurrence of antigen encounter by the cell of origin in chronic lymphocytic leukaemia (CLL) and CLL-like monoclonal B-cell lymphocytosis (MBL). However, the scenario in which this event might occur remains unknown. In order to gain insight into this scenario we investigated the molecular, cytogenetic and haematological features of 223 CLL-like (n = 84) and CLL (n = 139) clones with stereotyped (n = 32) versus non-stereotyped (n = 191) immunoglobulin heavy chain variable region (IGHV) amino acid sequences. Overall, stereotyped CLL-like MBL and CLL clones showed a unique IGHV profile, associated with higher IGHV1 and lower IGHV3 gene family usage (P = 0·03), longer IGHV complementary determining region 3 (HCDR3) sequences (P = 0·007) and unmutated IGHV (P 0·05), it did show a significant association with the presence of myelodysplastic syndrome (MDS)-associated immunophenotypes on peripheral blood neutrophils and/or monocytes (P = 0·01). Altogether our results point to the potential involvement of different selection forces in the expansion of stereotyped vs. non-stereotyped CLL and CLL-like MBL clones, the former being potentially favoured by an underlying altered haematopoiesis.Funded by: Red Temática de Investigación Cooperativa en Cáncer (RTICC) of Instituto de Salud Carlos III – FONDOS FEDER. Grant Numbers: RD06/0020/0035, RD12/0036/0048, FIS PI06/0824-FEDER, PS09/02430-FEDER; FIS PI12/00905-FEDER
Fondo de Investigación Sanitaria of Instituto de Salud Carlos III. Grant Numbers: GRS206/A/08, SAN/1778/2009; Gerencia Regional de Salud, Consejería de Educación and Consejería de Sanidad of Castilla y León. Grant Numbers: FS/1-2010, FS/19-2013;
Fundación Memoria D. Samuel Solórzano, Universidad de Salamanca; Fundação para a Ciência e Tecnologia of Portugal. Grant Number: SFRH/BD/31609/2006; Fundación Científica de la Asociación Española contra el Cáncer. Grant Number: AECC-2008.Peer Reviewe
Low-count monoclonal B-cell lymphocytosis persists after seven years of follow up and is associated with a poorer outcome
Low-count monoclonal B-cell lymphocytosis is defined by the presence of very low numbers of circulating clonal B cells, usually phenotypically similar to chronic lymphocytic leukemia cells, whose biological and clinical significance remains elusive. Herein, we re-evaluated 65/91 low-count monoclonal B-cell lymphocytosis cases (54 chronic lymphocytic leukemia-like and 11 non-chronic lymphocytic leukemia-like) followed-up for a median of seven years, using high-sensitivity flow cytometry and interphase fluorescence in situ hybridization. Overall, the clone size significantly increased in 69% of low-count monoclonal B-cell lymphocytosis cases, but only one subject progressed to high-count monoclonal B-cell lymphocytosis. In parallel, the frequency of cytogenetic alterations increased over time (32% vs. 61% of cases, respectively). The absolute number of the major T-cell and natural killer cell populations also increased, but only among chronic lymphocytic leukemia-like cases with increased clone size vs. age- and sex-matched controls. Although progression to chronic lymphocytic leukemia was not observed, the overall survival of low-count monoclonal B-cell lymphocytosis individuals was significantly reduced vs. non-monoclonal B-cell lymphocytosis controls (P=0.03) plus the general population from the same region (P≤0.001), particularly among females (P=0.01); infection and cancer were the main causes of death in low-count monoclonal B-cell lymphocytosis. In summary, despite the fact that mid-term progression from low-count monoclonal B-cell lymphocytosis to high-count monoclonal B-cell lymphocytosis and chronic lymphocytic leukemia appears to be unlikely, these clones persist at increased numbers, usually carrying more genetic alterations, and might thus be a marker of an impaired immune system indirectly associated with a poorer outcome, particularly among females.This work was supported by the RD06/0020/0035 and RD12/0036/0048 grants from Red Temática de Investigación Cooperativa en Cáncer (RTICC), Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, (Madrid, Spain and
FONDOS FEDER); CB16/12/00400 grant (CIBERONC, Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, (Madrid, Spain and FONDOS FEDER); the FIS PI06/0824-FEDER, PS09/02430-FEDER, PI12/00905- FEDER, DTS15/00119-FEDER, PI16/00787-FEDER and PI17/00399-FEDER grants, from the Fondo de Investigación Sanitaria of Instituto de Salud Carlos III; the GRS206/A/08 grant, (Ayuda al Grupo GR37 de Excelencia, SAN/1778/2009) from the Gerencia Regional de Salud (Consejería de Educación and Consejería de Sanidad of Castilla y León, Valladolid, Spain) and the SA079U14 grant (Consejería de Educación and Consejería de Sanidad of Castilla y León, Valladolid, Spain). ML Gutiérrez is supported by grant PTA2014-09963-I from the Instituto de Salud Carlos III
Combined Patterns of IGHV Repertoire and Cytogenetic/Molecular Alterations in Monoclonal B Lymphocytosis versus Chronic Lymphocytic Leukemia
Background:Chronic lymphocytic leukemia (CLL)-like monoclonal B lymphocytosis (MBL) with (MBLhi) or without (MBLlo) absolute B-lymphocytosis precedes most CLL cases,the specific determinants for malignant progression remaining unknown.Methodology/Principal Findings:For this purpose, simultaneous iFISH and molecular analysis of well-established cytogenetic alterations of chromosomes 11, 12, 13, 14 and 17 together with the pattern of rearrangement of the IGHV genes were performed in CLL-like cells from MBL and CLL cases. Our results based on 78 CLL-like MBL and 117 CLL clones from 166 subjects living in the same geographical area, show the existence of three major groups of clones with distinct but partially overlapping patterns of IGHV gene usage, IGHV mutational status and cytogenetic alterations. These included a group enriched in MBLloclones expressing specific IGHV subgroups (e.g. VH3-23) with no or isolated good-prognosis cytogenetic alterations, a second group which mainly consisted of clinical MBLhiand advanced stage CLL with a skewed but different CLL-associated IGHV gene repertoire (e.g. VH1-69), frequently associated with complex karyotypes and poor-prognosis cytogenetic alterations, and a third group of clones with intermediate features, with prevalence of mutated IGHV genes, and higher numbers of del(13q)+clonal B-cells.Conclusions/Significance:These findings suggest that the specific IGHV repertoire and IGHV mutational status of CLL-like B-cell clones may modulate the type of cytogenetic alterations acquired, their rate of acquisition and/or potentially also their clinical consequences. Further long-term follow-up studies investigating the IGHV gene repertoire of MBLloclones in distinct geographic areas and microenvironments are required to confirm our findings and shed light on the potential role of some antigen-binding BCR specificities contributing to clonal evolution
El grupu neandertal de la Cueva d'El Sidrón (Borines, Piloña).
Na monografía clásica de Puig y Larraz (1896: 250-252) amiéntense delles cavidaes del Conceyu de Piloña2 , pero non la Cueva d’El Sidrón (Fig. 1). Esta conocíase, ensin dulda, dende la Guerra Civil y el maquis al servir d’abellugu a persiguíos políticos, y guarda una alcordanza imborrable nuna de les sos múltiples entraes, yá qu’ellí ta enterrada Olvido Otero González (1908-1938). Per El Sidrón pasaron munches persones a lo llargo de los años, pero en 1994 prodúxose’l descubrimientu per parte d’unos espeleólogos xixoneses d’unos güesos humanos que dieron un importante xiru a la conocencia de los nuesos antepasaos neandertale
El grupu neandertal de la Cueva d' El Sidrón (Borines, Piloña)
El artículo está en la lengua asturianaPeer reviewe
Unexplored olive cultivars from the Valencian Community (Spain): some chemical characteristics as a valorization strategy
[EN] The olive processing industry has till date been dominated by a small group of cultivars, leading to the possibility of some olive cultivars becoming extinct in the near future. In this study, we determined the composition of some chemical components in the olive oils from 31 minor olive cultivars of the Valencian Community. Our main aim was to identify suitable cultivars, which could produce differentiated olive oils, thus aiming towards their valorization. The average oil content of minor olive cultivars was found to be good, with some of them reporting approximately 60% (dry basis). On average, the total phenolic content was 229mg kg(-1), with cv. Mas Blanc reporting the highest content (570mg kg(-1)). Among the various tocopherols found in olives, -tocopherol was the main constituent, with a maximum concentration of 290.6mg kg(-1). Linoleic acid was the main polyunsaturated fatty acid and varied between 3.4% (cv. Del Pomet) and 16.9% (cv. Blanqueta Enguera). Special attention needs to be paid to the composition of sterols, since some olive oils exceeded the limits established for some sterols by the current European legislation. Some of the cultivars studied were highly productive, and originated differentiated olive oils with a rich composition of antioxidants and essential fatty acids. In some cases, these beneficial compounds were higher than those of commercial oils obtained from the most common cultivars worldwide. These results could contribute to the commercial exploitation of some of the studied cultivars.Salazar-García, DC.; Malheiro, R.; Pereira, JA.; López- Cortés, I. (2019). Unexplored olive cultivars from the Valencian Community (Spain): some chemical characteristics as a valorization strategy. 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