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Infections were performed with Streptococcus pneumoniae (5 105 CFU) or Staphylococcus aureus (5 105 CFU) from an equal volume of bacterial suspension in sterile saline, using an Imject ThermoJet (Termo, France). Before injection, the viability of the bacteria was determined by plating serial dilutions of each bacteria on brain-heart infusion agar. 48 h after the bacterial infection, the animals were anesthetised and sacrificed.


Prospectively, the weight range in which each C-reactive protein (CRP) value was tested was set at 0.5 to 50 mg/dL in patients with cardiac disease and from 0 to 15 mg/dL in patients with chronic kidney disease (CKD). Given that patients with diabetes, hypertension and multiple cardiovascular disease (CVD) exhibit a higher CRP level than the general population, the target range was set at 1 to 300 mg/dL in patients with diabetes, 0 to 100 mg/dL in patients with hypertension and CKD, and 1 to 50 mg/dL in patients with CVD or valvular disease. Moreover, the CRP level of asymptomatic healthy individuals who were undergoing routine health screening was tested up to a target value of 10 mg/dL. Of the entire cohort, 40 patients (15%), and 31 patients (14%) were excluded from the study. The inclusion and exclusion criteria were applied as follows:


Splenocytes from the different experimental groups were isolated and cultured in the presence of anti-CD3 (1g) anti-CD28 (2g) antibodies for 24 h. Ficoll-Paque density gradient centrifugation and cell-sorting (MoFlo Astrios, Beckman Coulter, France) was used to isolate CD4+CD44+ and CD8+CD44+ T-cells. 3d9ccd7d82






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