Emerging role of monocytes and their intracellular calcium pattern in spondyloarthritis.

Spondyloarthritis (SpA) comprises multifactorial diseases characterized by a complex interplay between an inherited background and environmental factors that lead to immune response dysregulation and inflammation. Unlike for other rheumatic diseases, no specific biomarkers are available in clinical practice for diagnosing SpA. The aim of the present study was to search new potential biomarkers for SpA diagnosis by focusing on the innate immune response. An evaluation was made of the mRNA expression levels of inflammatory cytokines (TNF-α, IL-1ß, TGF-ß1, S100A8, S100A9) and matrix metalloproteinases (MMP3, MMP8, MMP9) in blood mononuclear cells of SpA patients (n = 64) with respect to controls (n = 100). In parallel, the pattern of intracellular calcium flows of blood monocytes was verified in order to ascertain whether any specific fingerprint characterizes innate immune cells in SpA patients. Inflammatory cytokines and MMPs expression levels were not correlated with SpA, while in this disease a reduced expression of the S100A8 and a decreased frequency of monocytes showing intracellular calcium flows were observed. In conclusion, no specific signs of systemic inflammation are detectable in SpA, but the disease affects the "on-off" mechanisms that regulate the concentration of intracellular calcium and calcium-related proteins. This potentially pave the way for the discovery of new biomarkers.

Relevance of pre-analytical blood management on the emerging cardiovascular protein biomarkers TWEAK and HMGB1 and on miRNA serum and plasma profiling.

BACKGROUND: Disease-independent sources of biomarker variability include pre-analytical, analytical and biological variance. The aim of the present study was to evaluate whether the pre-analytical phase has any impact on the emerging heart disease TWEAK and HMGB1 protein markers and miRNA biomarkers, and whether peptidome profiling allows the identification of pre-analytical quality markers. METHODS: An assessment was made of sample type (serum, EDTA-Plasma, Citrate-Plasma, ACD-plasma, Heparin-plasma), temperature of sample storage (room temperature or refrigerated), time of sample storage (0.5, 3, 6 and 9h) and centrifugation (one or two-step). Aliquots of all processed samples were immediately frozen (-80°C) before analysis. Proteins were assayed by ELISAs, miRNA expression profile by microarray and peptidome profiling by MALDI-TOF/MS. RESULTS: Temperature, time and centrifugation had no impact on TWEAK and HMGB1 results, which were significantly influenced by matrix type, TWEAK levels being significantly higher (F=194.7, p<0.0001), and HMGB1 levels significantly lower (F=36.32, p<0.0001) in serum than in any other plasma type. Unsuitable miRNA results were obtained using Heparin-plasma. Serum miRNA expression profiles depended mainly on temperature, while EDTA-plasma miRNA expression profiles were strongly affected by the centrifugation method used. MALDI-TOF/MS allowed the identification of seven features as indices of pre-analytical serum (m/z at 1206, 1350, 1865 and 2021) or EDTA-plasma (m/z 1897, 2740 and 2917) degradation. CONCLUSIONS: Serum and EDTA-plasma allow the analysis of both proteins and miRNA emerging biomarkers of heart diseases. Refrigerated storage prevents an altered miRNA expression profile also in cases of a prolonged time-interval between blood drawing and processing.

Erythropoietin therapy and acute preoperative normovolaemic haemodilution in infants undergoing craniosynostosis surgery.

BACKGROUND: A retrospective study was performed to evaluate whether pretreatment with erythropoietin and iron combined with acute preoperative normovolaemic haemodilution (APNH) could decrease homologous blood transfusion in craniosynostosis (CS) surgery. A treated group was compared with a historical group of infants who underwent surgery with no pretreatment. METHODS: The charts of 25 healthy infants who underwent CS surgery were reviewed. Nine of them underwent surgery with no treatment beforehand. Sixteen infants were given erythropoietin at a dosage of 300 U.kg -1 two times per week and iron (elemental iron 10 mg.kg-1.day-1) for 3 weeks before surgery. On the day of surgery APNH was performed after induction of general anaesthesia; a precalculated amount of autologous blood was withdrawn and replaced by hydroxyethyl starch 6%. RESULTS: Eleven of the 16 infants of the study group received only autologous blood. Five of 16 received homologous blood transfusion vs seven of nine infants in the control group. CONCLUSIONS: APNH combined with erythropoietin was effective in reducing homologous blood requirements during CS surgery. Further studies are necessary on a larger scale to assess the role of this technique in avoiding homologous blood transfusion and to evaluate how infants can benefit from this combined approach.