ABSTRACT
The level of expression of housekeeping genes is in general considered stable, and a representative gene such as glyceraldehyde-3-phosphate dehydrogenase is commonly used as an internal control for quantitating mRNA. However, expression of housekeeping genes is not always constant under pathological conditions. To determine which genes would be most suitable as internal controls for quantitative gene expression studies in human liver diseases, we quantified 12 representative housekeeping genes in 27 non-cancerous liver tissues (normal, chronic hepatitis C with and without liver cirrhosis). We identified ß-glucuronidase as the most suitable gene for studies on liver by rigorous statistical analysis of inter- and intra-group comparisons. We conclude that it is important to determine the most appropriate control gene for the particular condition to be analyzed.
Subject(s)
Gene Expression Profiling/methods , Genes, Essential/genetics , Glucuronidase/genetics , Liver Diseases/genetics , RNA, Messenger/analysis , Humans , Real-Time Polymerase Chain ReactionABSTRACT
Indenyl-functionalised BEt(3)-adduct NHCs were prepared by the reaction of imidazolium pro-ligands with LiBEt(3)H. This compound was converted into the indenyl-coordinate molybdenum complex dangling the NHC·BEt(3) moiety as a substituent. The stepwise coordination of NHC afforded a chelate-type mononuclear complex as well as a bimetallic complex.
Subject(s)
Boranes/chemistry , Heterocyclic Compounds/chemistry , Indenes/chemistry , Methane/analogs & derivatives , Molybdenum/chemistry , Organometallic Compounds/chemical synthesis , Boranes/chemical synthesis , Crystallography, X-Ray , Heterocyclic Compounds/chemical synthesis , Imidazoles/chemistry , Ligands , Methane/chemistry , Models, Molecular , Molecular Structure , Organometallic Compounds/chemistry , StereoisomerismABSTRACT
We studied the incidence of t(14;18)(q32;q21) in 54 patients with follicular lymphoma (FL) by dual-color fluorescence in situ hybridization on paraffin-embedded tissue sections (tissue-FISH) using probes for BCL2 and immunoglobulin heavy chain (IGH) genes. The t(14;18) was detected in 28 (56%) of 50 patients, who were successfully analyzed. On the other hand, BCL2 protein expression was detected in 45 (83%) of 54 patients evaluated by immunohistochemical staining. There was a discrepancy between t(14;18) and BCL2 expression. The absence of t(14;18) was associated with disease-free survival (P=0.02). There was no statistical difference, however, in overall survival and failure-free survival between the t(14;18)-positive and -negative groups. Tissue-FISH should be of great value to detect t(14;18) in FL because this method enabled us to demonstrate specifically t(14;18)-positive individual cells in neoplastic follicles on paraffin-embedded tissue sections. Furthermore, tissue-FISH can be applied to small specimens obtained from endoscopic biopsy or specimens obtained more than 10 years ago. We validated the usefulness of tissue-FISH as a diagnostic procedure and retrospective meta-analysis for malignant lymphoma.
