In-Depth Proteomic Characterization of Classical and Non-Classical Monocyte Subsets.

Monocytes are bone marrow-derived leukocytes that are part of the innate immune system. Monocytes are divided into three subsets: classical, intermediate and non-classical, which can be differentiated by their expression of some surface antigens, mainly CD14 and CD16. These cells are key players in the inflammation process underlying the mechanism of many diseases. Thus, the molecular characterization of these cells may provide very useful information for understanding their biology in health and disease. We performed a multicentric proteomic study with pure classical and non-classical populations derived from 12 healthy donors. The robust workflow used provided reproducible results among the five participating laboratories. Over 5000 proteins were identified, and about half of them were quantified using a spectral counting approach. The results represent the protein abundance catalogue of pure classical and enriched non-classical blood peripheral monocytes, and could serve as a reference dataset of the healthy population. The functional analysis of the differences between cell subsets supports the consensus roles assigned to human monocytes.

Detection of Missing Proteins Using the PRIDE Database as a Source of Mass Spectrometry Evidence.

The current catalogue of the human proteome is not yet complete, as experimental proteomics evidence is still elusive for a group of proteins known as the missing proteins. The Human Proteome Project (HPP) has been successfully using technology and bioinformatic resources to improve the characterization of such challenging proteins. In this manuscript, we propose a pipeline starting with the mining of the PRIDE database to select a group of data sets potentially enriched in missing proteins that are subsequently analyzed for protein identification with a method based on the statistical analysis of proteotypic peptides. Spermatozoa and the HEK293 cell line were found to be a promising source of missing proteins and clearly merit further attention in future studies. After the analysis of the selected samples, we found 342 PSMs, suggesting the presence of 97 missing proteins in human spermatozoa or the HEK293 cell line, while only 36 missing proteins were potentially detected in the retina, frontal cortex, aorta thoracica, or placenta. The functional analysis of the missing proteins detected confirmed their tissue specificity, and the validation of a selected set of peptides using targeted proteomics (SRM/MRM assays) further supports the utility of the proposed pipeline. As illustrative examples, DNAH3 and TEPP in spermatozoa, and UNCX and ATAD3C in HEK293 cells were some of the more robust and remarkable identifications in this study. We provide evidence indicating the relevance to carefully analyze the ever-increasing MS/MS data available from PRIDE and other repositories as sources for missing proteins detection in specific biological matrices as revealed for HEK293 cells.

Cardiovascular fitness in youth: association with obesity and metabolic abnormalities.

UNLABELLED: Therapies currently implemented for obesity are focused on nutritional aspects and on physical activity. In order to make physical activity a positive therapy instead of triggering disabilities it is relevant to accurately assess cardiovascular fitness. OBJECTIVE: To assess the cardiovascular fitness by measuring the peak oxygen consumption and to asses their relationship with classical cardiometabolic parameters. METHODS: A modified Balke protocol was applied to one hundred and twenty-six Caucasians (60% males), ranging between 9 and 16 years old, who underwent an assessment of obesity. The non-obese group consisted of healthy age and sex matched subjects who were invited to participate from the general population. RESULTS: Significant differences in consumption of oxygen peak between non-obese and obese individuals were observed. In contrast, no significant differences existed between the categories of obesity. Furthermore in obese subjects consumption of oxygen peak was inversely correlated with parameters of cardiometabolic risk, particularly insulin and HOMA index. In addition, two predictive equations of consumption of oxygen peak, with an R2 of 0.74 and 0.84, respectively, have been developed. CONCLUSION: The consumption of oxygen peak is a relevant clinical parameter that should be included in the routine clinical assessment of obese subjects. Therefore, it is crucial to make exercise tests more affordable which can be achieved by employing predictive equations.

Las terapias que se implantan actualmente para la obesidad se centran en los aspectos nutricionales y sobre la actividad física. Con el fin de hacer que la actividad física sea una terapia positiva en vez de un desencadenador de discapacidades, es relevante evaluar de forma precisa el entrenamiento cardiovascular. Objetivo: evaluar el entrenamiento cardiovascular midiendo el consumo máximo de oxígeno y evaluar su relación con los parámetros cardiometabólicos clásicos. Métodos: se aplicó el protocolo modificado de Balke a 126 individuos caucásicos (60 % de varones), con edades entre 9 y 16 años, que se sometieron a una evaluación de obesidad. El grupo de no obesos consistía de individuos sanos, de la población general, emparejados por edad y sexo y a los que se les invitó a participar. Resultados: se observaron diferencias significativas en el consumo máximo de oxígeno entre los indiviudos obesos y no obesos. Por contra, no existían diferencias significativas entre las categorías de obesidad. Además, en los sujetos obesos, el consumo máximo de oxígeno se correlacionó de forma inversa con los parámetros de riesgo cardiometabólico, particularmente con la insulina y el índice HOMA. Además, se han desarrollado dos ecuaciones predictivas del consumo máximo de oxígeno con una R2 de 0,74 y de 0,84, respectivamente. Conclusión: el consumo máximo de oxígeno es un parámetro clínico relevante que debería incluirse en la evaluación clínica rutinaria de los sujetos obesos. Por lo tanto, es crucial hacer que las pruebas de esfuerzo sean más asequibles, que puedan alcanzarse empleando las ecuaciones predictivas.

Cardiovascular fitness in youth; association with obesity and metabolic abnormalities / Entrenamiento cardiovascular en la juventud; asociación con obesidad y anomalías metabólicas

Therapies currently implemented for obesity are focused on nutritional aspects and on physical activity. In order to make physical activity a positive therapy instead of triggering disabilities it is relevant to accurately assess cardiovascular fitness. Objective: To assess the cardiovascular fitness by measuring the peak oxygen consumption and to asses their relationship with classical cardiometabolic parameters. Methods: A modified Balke protocol was applied to one hundred and twenty-six Caucasians (60% males), ranging between 9 and 16 years old, who underwent an assessment of obesity. The non-obese group consisted of healthy age and sex matched subjects who were invited to participate from the general population. Results: Significant differences in consumption of oxygen peak between non-obese and obese individuals were observed. In contrast, no significant differences existed between the categories of obesity. Furthermore in obese subjects consumption of oxygen peak was inversely correlated with parameters of cardiometabolic risk, particularly insulin and HOMA index. In addition, two predictive equations of consumption of oxygen peak, with an R2 of 0.74 and 0.84, respectively, have been developed. Conclusion: The consumption of oxygen peak is a relevant clinical parameter that should be included in the routine clinical assessment of obese subjects. Therefore, it is crucial to make exercise tests more affordable which can be achieved by employing predictive equations (AU)

Las terapias que se implantan actualmente para la obesidad se centran en los aspectos nutricionales y sobre la actividad física. Con el fin de hacer que la actividad física sea una terapia positiva en vez de un desencadenador de discapacidades, es relevante evaluar de forma precisa el entrenamiento cardiovascular. Objetivo: evaluar el entrenamiento cardiovascular midiendo el consumo máximo de oxígeno y evaluar su relación con los parámetros cardiometabólicos clásicos. Métodos: se aplicó el protocolo modificado de Balke a 126 individuos caucásicos (60 % de varones), con edades entre 9 y 16 años, que se sometieron a una evaluación de obesidad. El grupo de no obesos consistía de individuos sanos, de la población general, emparejados por edad y sexo y a los que se les invitó a participar. Resultados: se observaron diferencias significativas en el consumo máximo de oxígeno entre los individuos obesos y no obesos. Por contra, no existían diferencias significativas entre las categorías de obesidad. Además, en los sujetos obesos, el consumo máximo de oxígeno se correlacionó de forma inversa con los parámetros de riesgo cardiometabólico, particularmente con la insulina y el índice HOMA. Además, se han desarrollado dos ecuaciones predictivas del consumo máximo de oxígeno con una R2 de 0,74 y de 0,84, respectivamente. Conclusión: el consumo máximo de oxígeno es un parámetro clínico relevante que debería incluirse en la evaluación clínica rutinaria de los sujetos obesos. Por lo tanto, es crucial hacer que las pruebas de esfuerzo sean más asequibles, que puedan alcanzarse empleando las ecuaciones predictivas (AU)

Vincristine, doxorubicin, cyclophosfamide, actinomycin D, ifosfamide, and etoposide in adult and pediatric patients with nonmetastatic Ewing sarcoma. Final results of a monoinstitutional study.

AIMS AND BACKGROUND: To investigate a six-drug combination in patients with nonmetastatic Ewing sarcoma, focusing on chemotherapy-induced necrosis and chemotherapy toxicity in adult and pediatric patients. METHODS AND STUDY DESIGN: Alternating cycles of vincristine (1.5 mg/m2), doxorubicin (80 mg/m2) and cyclophosfamide (1200 mg/m2) (weeks 0, 6, 13, 22 and 31), ifosfamide (9 g/m2), vincristine (1.5 mg/m2), and actinomycin D (1.5 mg/m2) (weeks 3, 16, 25 and 34), and ifosfamide (9 g/m2) and etoposide (450 mg/m2) (weeks 9, 19, 28 and 37) were administered. Primary chemotherapy-induced necrosis was graded: G3 (complete necrosis), G2 (microfoci of tumor cells) and G1 (macrofoci of tumor cells). RESULTS: From 1996 to 1999, 50 patients with Ewing sarcoma were enrolled. The median age was 23.5 years (range, 4-56). Chemotherapy-induced necrosis (in 28 patients) was G3 in 36%, G2 in 21% and G1 in 43%. At a median follow-up of 110 months (range, 36-129), 5-year overall survival and event-free survival were 72% and 66%, respectively. According to histologic response, 5-year event-free survival was 90% in G3, 83% in G2, and 42% in G1 (P = 0.02). In adult and pediatric (<18 years) patients, the incidence of G4 leukopenia was 62% and 74%, respectively, with febrile neutropenia in 13% and 21%, respectively. G4 thrombocytopenia occurred in 3% of cycles in adults and in 7% in pediatric patients. Platelet and red blood cell transfusions were required respectively in 1% and 11% of cycles in adults and in 6% and 24% of cycles in pediatric patients. CONCLUSIONS: The six-drug combination can be administered safely in adult and pediatric populations. About 40% of patients have a poor chemotherapy-induced tumor necrosis, leading to poor probability of survival. New strategies are recommended to improve survival of poor responders to the six-drug combination.

Proteomic analysis of quorum sensing in Rhizobium leguminosarum biovar viciae UPM791.

Production of quorum-sensing signal molecules of the acyl-homoserine lactone (AHL) type by Rhizobium leguminosarum bv. viciae UPM791 is dependent on its plasmid content. Curing of two of its four native plasmids, pUPM791d and pSym, resulted in loss of production of the largest (C(14)) and the three smaller (C(6)-C(8)) AHLs, respectively. Introduction of a lactonase-containing plasmid resulted in AHL signal degradation and quorum quenching. The quorum-dependent proteome was studied in these strains by DIGE. Quorum quenching affected a small (1.7%) fraction of the detected spots in the wild-type and a smaller (0.6%) fraction in the pSym-cured strain. Unexpectedly, quorum quenching affected up to 3.3% of the detected spots in the pUPM791d-cured strain, suggesting that C(14)-AHL normally interferes with the quorum response mediated by other AHLs. This, together with the observation that ca. 50% of the quorum-regulated proteins in strain UPM791 showed AHL-mediated repression, suggests that an important part of their functionality can be exerted through repression, although AHLs are usually considered as gene expression inducers. The three main quorum-induced polypeptides were identified by MALDI-MS as charge isoforms of the rhizospheric RhiA protein. Another major quorum-induced polypeptide was only present in the pUPM791d-cured strain and could not be identified.