Advia 1650 analyser

Biochemical determinations and nucleated cell counting were performed by using an ADVIA^®1650 analyser (SIEMENS Healthcare Diagnostics S.L., Berlin, Germany) and an ADVIA^®2120 haematology counter (SIEMENS Healthcare Diagnostics S.L., Berlin, Germany), repectively. Serum TGF-β1 levels were measured using ELISA plates (Human TGF-beta1 Platinum ELISA; ref. BMS249/4TEN; eBioscience) following commercial guidelines. Optical densities were assayed at 450 nm (Labsystem Multiscan MS), and protein concentration was calculated from standard curves.

All laboratory variables were assessed upon admission to the geriatric rehabilitation unit. Serum leptin levels were assessed using a commercially available ELISA (Human Leptin Quantikine ELISA Kit (R&D Systems, Abingdon, UK)). The serum samples were stored at −20 °C prior to analysis. The serum albumin was measured using a BNII analyser (Siemens, Saint-Denis, France). C-reactive protein (CRP) was measured by immunoturbidimetry using an Advia 1650 analyser (Siemens Healthcare, Saint-Denis, France). An ultrasensitive CRP assay was not available at the time of the study. Immunological variables were assessed using flow cytometry immunophenotyping, as described previously [25 (link)]. Absolute peripheral blood CD4 and CD8 T-cell counts were determined using a Cyto-Stat tetraCHROME device (including labelling with CD45, CD3, CD4, and CD8) and acquisition on an FC500 flow cytometer (both from Beckman Coulter, Villepinte, France). Counts of naïve, memory, and terminal effector T-cells were determined as follows: CD8 and CD4 naïve T-cells were defined as CD45RA+CD62L+, peripheral memory T-cells were defined as CD45RA−CD62L−, and terminal effector CD8 T-cells were defined as CD45RA+CD62L−CD28−. The anti-CD62L antibody was obtained from BD Pharmingen (BD Biosciences, San Jose, CA, USA) and the CD4, CD8, and CD28 antibodies (Caltag) came from Life Technologies (Carlsbad, CA, USA).

Biochemical determinations were carried out by an ADVIA^®1650 analyser (SIEMENS Healthcare Diagnostics S.L., Berlin, Germany) while neuron-specific enolase (NSE) was measured with electrochemiluminescence analyser (MODULAR ANALYTICS Cobas E-601, Roche Diagnostics; Mannheim, Germany). The nucleated cell counting was performed using an ADVIA^®2120 hematology counter (SIEMENS Healthcare Diagnostics S.L., Berlin, Germany).
Serum sCD14 levels were measured by means of an enzyme-linked immunosorbant assay (ELISA) with the Quantikine®Human sCD14 Immunoassay kit (R&D Systems, MN, USA). Optical densities were recorded at 450 nm and protein concentration was calculated from standard curves.