The gene HIF1�� is composed of 15 exons, resulting in a principal transcript (HIF1��WT) [3,15] and seven alternative splice variants which have been reported in human cell lines [16-20]. Amplification of HIF1��WT showed that it was expressed by circulating blood cells, as well as the splicing variants HIF1��TAG and HIF1��736 (Figure (Figure1).1). HIF1��516 and selleck chemicals llc HIF1��557 splice variants tested two isoforms coding for negative dominants. These two isoforms were not or were poorly expressed by circulating blood cells. Relative expression of different isoforms was similar between patients and volunteers (Figure (Figure11).Figure 1Expression of different HIF1�� variants in shock patients (black bars) and controls (grey bars).In a subgroup of six patients, at H0, blood was collected from both arterial and venous lines.

With regard to the expression of HIF1a, no difference was found between the venous and arterial blood samples (data not shown). Then, a group of 11 healthy volunteers, non-smokers, was evaluated for HIF1�� expression and used as controls.Statistical analysisFrom previous studies [14,21], 44 patients with shock were required to achieve a predictive value of 90% with a bias <5% and a 5% risk ��. Data were analyzed using the software SPSS and R. Quantitative variables are expressed as median and interquartile range. Qualitative variables are expressed as absolute counts and percentages. Differences between groups were tested using non-parametric tests (Mann-Whitney and Kruskall-Wallis tests). A P level of 0.05 or less was considered significant.

ResultsPatient characteristicsFifty patients with shock (average age 57 (range: 18 to 80 years) and 11 healthy volunteers (average age 50 (range: 29 to 70 years) were prospectively included. Women represented 25% of the cohort of patients and 27% of the cohort of healthy volunteers. The HIF1�� expression was unaffected by sex (P = 0.7) or age (P = 0.8). The causes of shock were sepsis, bleeding, and cardiac dysfunction in 39 (78%), 9 (18%), and 2 (4%) cases, respectively (Table (Table1).1). Plasma bilirubin concentration and SOFA score differed significantly in survivors and non-survivors (Table (Table11).Table 1Characteristics of the patients according to their survival (day 28).HIF1�� expressionAt any time points of the study period, the expression of HIF1�� was significantly increased in the patients with shock (Figure (Figure2).

2). At H0, 121 (range: 72 to 168) normalized copies were found in patients with shock, as compared with 46 (range: 38 to 54) normalized copies in healthy volunteers (P <0.01). The detailed values for each time point are presented in Batimastat Table Table2.2. Of note, the expression of HIF1�� did not differ according to the type of shock (data not shown). We did not find a relation between the expression of HIF1�� and the absolute number of white blood cells (data not shown).