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27 results on '"Fernández-Segoviano, Pilar"'

11. Transcriptional profiling and genotyping of degraded nucleic acids from autopsy tissue samples after prolonged formalin fixation times

Author

Ferruelo, Antonio, El-Assar, Mariam, Lorente, José A, Nin, Nicolás, Peñuelas, Oscar, Fernández-Segoviano, Pilar, Gonzalez, Constancio, and Esteban, Andrés

Subjects
Time Factors, Tissue Fixation, Genotype, Tumor Necrosis Factor-alpha, Gene Expression Profiling, RNA Stability, DNA, Peptidyl-Dipeptidase A, Fixatives, Liver, Formaldehyde, Humans, RNA, Original Article, Autopsy, Artifacts, Promoter Regions, Genetic
Abstract

Background: Samples used for genotyping and transcription studies are obtained and conserved in very specific conditions. The possibility to use autopsy tissue samples, which contain nucleic acids of very poor quality, would open new possibilities for genetic studies. Methods: We have used liver tissue samples from autopsy cases to (i) determine its quality; (ii) study gene expression of 13 genes involved in different cell processes, before and after cDNA pre-amplification (quantitative reverse transcriptase polymerase chain reaction); and (iii) analyze the presence of 2 common polymorphisms of relevance for illness (ACE I/D genotype by PCR amplification, and TNF-α promoter gene polymorphism, by DNA sequencing). Results: Samples were grouped according to different buffered formalin fixation times (group 1

Published
2011

13. A Metabolomic Approach to the Pathogenesis of Ventilator-induced Lung Injury

Author

Izquierdo-García, José L., primary, Naz, Shama, additional, Nin, Nicolás, additional, Rojas, Yeny, additional, Erazo, Marcela, additional, Martínez-Caro, Leticia, additional, García, Antonia, additional, de Paula, Marta, additional, Fernández-Segoviano, Pilar, additional, Casals, Cristina, additional, Esteban, Andrés, additional, Ruíz-Cabello, Jesús, additional, Barbas, Coral, additional, and Lorente, José A., additional

Published
2014
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15. Clinical diagnoses and autopsy findings

Author

Tejerina, Eva, primary, Esteban, Andrés, additional, Fernández-Segoviano, Pilar, additional, María Rodríguez-Barbero, José, additional, Gordo, Federico, additional, Frutos-Vivar, Fernando, additional, Aramburu, José, additional, Algaba, Ángela, additional, Gonzalo Salcedo García, Óscar, additional, and Lorente, José A., additional

Published
2012
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26. Transcriptional profiling and genotyping of degraded nucleic acids from autopsy tissue samples after prolonged formalin fixation times.

Author

Ferruelo A, El-Assar M, Lorente JA, Nin N, Peñuelas O, Fernández-Segoviano P, Gonzalez C, and Esteban A

Subjects
Artifacts, Autopsy, Genotype, Humans, Liver chemistry, Peptidyl-Dipeptidase A genetics, Promoter Regions, Genetic, RNA Stability, Time Factors, Tissue Fixation methods, Tumor Necrosis Factor-alpha genetics, DNA chemistry, Fixatives chemistry, Formaldehyde chemistry, Gene Expression Profiling methods, RNA chemistry
Abstract

Background: Samples used for genotyping and transcription studies are obtained and conserved in very specific conditions. The possibility to use autopsy tissue samples, which contain nucleic acids of very poor quality, would open new possibilities for genetic studies., Methods: We have used liver tissue samples from autopsy cases to (i) determine its quality; (ii) study gene expression of 13 genes involved in different cell processes, before and after cDNA pre-amplification (quantitative reverse transcriptase polymerase chain reaction); and (iii) analyze the presence of 2 common polymorphisms of relevance for illness (ACE I/D genotype by PCR amplification, and TNF-α promoter gene polymorphism, by DNA sequencing)., Results: Samples were grouped according to different buffered formalin fixation times (group 1, <15 days; group 2, 60-90 days; group 3, 150-180 days; group 4, 240-270 days). Nucleic acids showed a time-dependent degradation. The expression of 13 genes could be studied in all cases from groups 1 and 2, only 7 from group 3 and none from group 4. cDNA preamplification allowed the study of all genes in all samples. DNA genotyping for ACE and TNF-α promoter region was possible in all cases., Conclusions: We conclude that nucleic acids extracted from autopsy specimens after prolonged periods of time in formalin were of sufficient quality to study gene expression and genotyping using currently available methodology and cDNA pre-amplification.

Published
2011

27. Large tidal volume mechanical ventilation induces vascular dysfunction in rats.

Author

Nin N, Valero JA, Lorente JA, de Paula M, Fernández-Segoviano P, Sánchez-Ferrer A, and Esteban A

Subjects
Analysis of Variance, Animals, Lung pathology, Male, Photomicrography, Random Allocation, Rats, Rats, Sprague-Dawley, Respiration, Artificial methods, Tidal Volume, Vascular Diseases pathology, Aorta, Thoracic pathology, Lung blood supply, Respiration, Artificial adverse effects, Vascular Diseases etiology
Abstract

Background: Experimental studies have shown that mechanical ventilation using high tidal volumes (V(T)) damages the lungs, causing pulmonary edema. We tested the hypothesis that high V(T) ventilation in rats induces major vascular dysfunction., Methods: Healthy Sprague-Dawley rats, weighing (mean +/- SD) 340 +/- 15 g, were ventilated with either V(T) = 9 mL/kg and positive end-expiratory pressure (PEEP) = 8 (n = 8) or V(T) = 35 mL/kg and PEEP = 0 (n = 8). The high V(T) used in the injurious ventilation group is in the V(T) range used in other studies to induce lung damage in a short period of time in rats. Lungs were removed for examination under light microscopy and vascular rings from the thoracic aorta were studied for isometric tension recording., Results: Relaxations to acetylcholine (p < 0.001) and sodium nitroprusside (p < 0.05) and contractions to norepinephrine were markedly decreased (p < 0.001) in the high V(T) group, as compared with the low V(T) group., Conclusion: Injurious mechanical ventilation in normal rats is associated with vascular dysfunction characterized by decreased relaxation to an endothelium-dependent vasodilator and to a nitrous oxide donor and by decreased response to norepinephrine.

Published
2005

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