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Masvidal L.,Human Molecular Genetics Group | Alvarez A.,Hospital Vall dHebron | Ruano L.,Hospital Vall dHebron | de Gracia J.,Hospital Vall dHebron | And 2 more authors.
Journal of Cystic Fibrosis | Year: 2012

Background: CFTR expression studies contribute in understanding the relationship between CFTR transcripts and clinical outcomes. Normalization of qPCR data is an essential step to determine target gene expression. Consequently, appropriate reference genes must be selected for each gene/tissue. In this work, we have assessed the suitability of four potential reference genes for CFTR expression analysis in nasal epithelium. Methods: B2M, GUSB, HPRT1 and ATP2B4 expression was evaluated in nasal epithelium samples (CFTR-wt controls, n. =21; CFTR-splicing group, n. =18) by RT-qPCR. Calibration curves were built and different analyses (geNorm, NormFinder, Mann-Whitney) were performed to evaluate gene expression stability between samples as well as between groups. Results and conclusions: We have applied an accurate approach to select reference genes for CFTR expression analysis in nasal epithelium. From the four genes assessed, GUSB and ATP2B4 have been validated as a reliable gene combination for CFTR gene qPCR data normalization. © 2012 European Cystic Fibrosis Society..


Luque A.,Hospital Universitari Mutua Terrassa | Luque A.,Cardiovascular Research Center | Luque A.,Human Molecular Genetics Group | Turu M.M.,Cardiovascular Research Center | And 6 more authors.
Frontiers in Bioscience - Elite | Year: 2012

Chlamydia pneumoniae (Cpn) could play an important role in the development of atherosclerosis. Cpn interferes with HIF-1alpha regulation in infected host cells during intracellular replication in hypoxia. We obtained carotid artery specimens with low (n=38), high (n=25) levels of stenosis and 10 control middle cerebral arteries. Fifty eight percent of the carotids with low levels of stenosis showed evidence of the viable Cpn. Ninety one percent of the positive results were derived from preatheromatous lesions. Only 12 percent of plaques removed at endarterectomy showed the presence of Cpn DNA. All control middle cerebral arteries failed to show evidence of live Cpn. Ninety one percent of sera from 22 endarterectomy patients showws the presence of Cpn antibodies. Immunohistology of carotid arteries with low levels of stenosis was used to confirm the presence of HIF-1alpha in infected specimens and showed a correlation between the over-expression of HIF-1alpha and Cpn in the plaque (p less than 0.05). Cpn might play an important role in activation and development of the initial stages of atherosclerotic lesions.


PubMed | Human Molecular Genetics Group
Type: Journal Article | Journal: Journal of cystic fibrosis : official journal of the European Cystic Fibrosis Society | Year: 2012

CFTR expression studies contribute in understanding the relationship between CFTR transcripts and clinical outcomes. Normalization of qPCR data is an essential step to determine target gene expression. Consequently, appropriate reference genes must be selected for each gene/tissue. In this work, we have assessed the suitability of four potential reference genes for CFTR expression analysis in nasal epithelium.B2M, GUSB, HPRT1 and ATP2B4 expression was evaluated in nasal epithelium samples (CFTR-wt controls, n=21; CFTR-splicing group, n=18) by RT-qPCR. Calibration curves were built and different analyses (geNorm, NormFinder, Mann-Whitney) were performed to evaluate gene expression stability between samples as well as between groups.We have applied an accurate approach to select reference genes for CFTR expression analysis in nasal epithelium. From the four genes assessed, GUSB and ATP2B4 have been validated as a reliable gene combination for CFTR gene qPCR data normalization.

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