IL8 and EDEM3 gene expression ratio indicates peripheral blood mononuclear cell (PBMC) quality.

BACKGROUND: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times. RESULTS: We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.

A new transcript splice variant of the human glucocorticoid receptor: identification and tissue distribution of hGR Delta 313-338, an alternative exon 2 transactivation domain isoform.

All human glucocorticoid receptor (hGR) isoforms are encoded by the NR3C1 gene consisting of seven core exons (exons 2-8) common to all protein isoforms. The gene has two major exon 8-9 splice variants and a 5'-UTR consisting of 11 alternative splice variants. The N-terminal region of the hGR includes a tau 1 transactivation domain that interacts with proteins in the basal transcriptional apparatus, including the TATA box-binding protein. Here, we report the existence and the tissue distribution of a novel splice variant, hGRDelta313-338, with a 26 residue (78 bp) deletion in this N-terminal region encoded by exon 2, between amino acids 313 and 338. The hGRDelta313-338 observed at the mRNA level represents a transcript variant encoding a smaller protein isoform detected by WB with a predicted deletion between the tau 1 domain and the DNA-binding domain (DBD) encoded by exons 3 and 4. Previous studies in transgenic mice showed that the removal of the entire exon 2 covering both the tau 1 transactivation domain and our deleted region produced a functional receptor albeit with an altered glucocorticoid-induced gene transcription pattern. Interestingly, the deleted residues show a number of potential phosphorylation sites including serine 317, known to be phosphorylated. It is thought that phosphorylation plays an important role in transactivation action of hGR. Thus, we hypothesize that hGRDelta313-338 represents a hGR isoform with an altered glucocorticoid-induced transactivation profile.

Thyroid cancer in Luxembourg: a national population-based data report (1983-1999).

BACKGROUND: Twenty years after the nuclear accident in Chernobyl (Eastern Europe), there is still a controversial debate concerning a possible effect of the radioactive iodines, especially I-131, on the increase of thyroid carcinomas (TCs) in Western Europe. Time trends in incidence rates of TC in Luxembourg in comparison with other European countries and its descriptive epidemiology were investigated. METHODS: The population-based data of the national Morphologic Tumour Registry collecting new thyroid cancers diagnosed between 1983 and 1999 at a nation-wide level in the central division of pathology were reviewed and focused on incidence rates of TC. Data from 1990 to 1999 were used to evaluate the distribution by gender, age, histological type, tumour size and the outcome. RESULTS: Out of 310 new thyroid carcinomas diagnosed between 1990 and 1999, 304 differentiated carcinomas (A: 80% papillary; B: 14.5% follicular; C: 3.5% medullary) and 6 anaplastic/undifferentiated TCs (D: 2%) were evaluated. The M/F-ratio was 1:3.2, the mean age 48.3 years (range: 13-92). The overall age-standardized (world population) incidence rates over the two 5-year periods 1990-1994 and 1995-1999 increased from 7.4 per 100,000 to 10.1 per 100,000 in females, from 2.3 per 100,000 to 3.6 per 100,000 in males. Only 3 patients were children or adolescents (1%), the majority of the patients (50%) were between 45 and 69 years of age. The percentage of microcarcinomas (<1 cm) was A: 46.4%, (115/248); B: 13.3%, (6/45); C: 27.3%, (3/11). The unexpected increase of TCs in 1997 was mainly due to the rise in the number of microcarcinomas. The observed 5-year survival rates for both genders were A: 96.0+/-2%; B: 88.9%; C: 90.9%; D: 0%. Prognosis was good in younger patients, worse in males and elderly, and extremely poor for undifferentiated TCs. CONCLUSION: The increasing incidence rates of TC, especially of the papillary type, seem mainly due to a rise in diagnosed microcarcinomas due to some extent to a change in histologic criteria and to more efficient diagnostic tools. This rise appears to be independent of the number of surgical treatments, the immigration rate, and the Chernobyl fallout as the incidence of TC in children remained stable.