Early inflammatory response in polytraumatized patients: Cytokines and heat shock proteins. A pilot study.

INTRODUCTION: In the initial phases after polytrauma there is an hyperinflammatory state that sometimes leads to multiple organ dysfunction syndrome (MODS) and death, and that appears to be responsible for posttraumatic immunosuppression; among the trigger endogenous stimuli, heat shock proteins (HSPs) have been proposed. The objectives of this study were to analyze if some inflammatory mediators can be considered prognostic biomarkers of outcome, and the possible role of HSPA1A in posttraumatic immunosuppression. HYPOTHESIS: Cytokines and HSPs could be early prognostic biomarkers of inflammatory and immune response in polytrauma patients. MATERIALS AND METHODS: A prospective observational descriptive pilot study was carried out, evaluating the early inflammatory and stress response of 18 polytraumatized patients (ISS>16) on hospital admission, at 12hours, 24hours, and 48hours posttrauma. Variable means were compared using non-parametric tests; qualitative and quantitative variables were analyzed using a Spearman's correlation test. RESULTS: Seven patients met criteria for MODS. Statistically significant changes were recorded in leukocyte count, C-reactive-protein (CRP), IL-6, TNF-α, and IL-1ß concentrations. HSPA1A levels were significantly higher immediately after the accident followed by gradual lowering. Anti-Hsp70 antibodies showed a significant reduction in all the studied time-points. MODS did not influence either plasma levels of leukocytes, fibrinogen, RCP or anti-Hsp70, but patients with MODS had higher plasma levels of IL-6 and TNF-α and a slower decrease of HSPA1A concentrations. DISCUSSION: The higher serum concentrations of TNF-α and IL-6 found in patients with MODS, suggests a possible role as potential early predictive markers for systemic inflammatory response and clinical complications. The higher levels of HSPA1A in patients with MODS, allows proposing HSPA1A as a useful prognostic trauma biomarker early after severe injury and to consider a "damage control surgery". The significant reduction in the levels of anti-Hsp70 antibodies could reflect a part of posttraumatic immunosuppression and hydrocortisone treatment might be suggested. LEVEL III: case-control study.

Peripheral leukocyte response to oncological radiotherapy: expression of heat shock proteins.

PURPOSE: To study Heat Shock Proteins (HSP) expression in patients subjected to radiotherapy and their potential use as biomarkers for radiation tolerance. An evaluation is also made of whether irradiated volume is critical to the outcome of normal tissue injury using polymorphonuclear neutrophils as biosensors, and whether HSP antibodies (Ab) may be involved in post-radiotherapy disease. MATERIAL AND METHODS: Twelve patients receiving the same total dose of radiotherapy, but in three different volumes, and four healthy volunteers used as controls were analysed. hsp27 and 70i mRNA were determined by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Southern-blot, HSP by flow cytometry, and HSP-Ab by Enzyme-linked Immnoadsorbent Assay (ELISA). The clinical protocol included radiation related toxicity based on clinical and analytical scales. RESULTS: Radiotherapy caused hsp downregulation, maximum in patients with the largest irradiated volumes, and a decrease in intracellular HSP content. Patients with greatest intraleukocyte HSP levels before treatment suffered more severe radiation morbidity. Patients with endocrine neoplasms presented the highest HSP-Ab titers. CONCLUSIONS: Radiotherapy downregulates hsp27 and 70i, which would enhance radiosensitivity. HSP content prior to treatment is suggested as a prognostic biomarker for radiation tolerance, with circulating leukocytes as biosensors. HSP-Ab may be biomarkers of tumor disease, but do not seem to be involved in the morbidity of acute post-radiotherapy disease, which is closely related to the volumes irradiated.

Fluorescent and compositional changes in crystallin supramolecular structures in pig lens during development.

Water soluble proteins (WSPs) in Sus scrofa lenses from pigs in different developmental stages: (young (GI), young adult (GII), and middle-aged (GIII)) were separated using GF-HPLC, yielding fractions of different molecular weights. Non-tryptophan (345/420 nm) and tryptophan (280/345 nm) fluorescence was measured in these fractions. Relative non-tryptophan fluorescence increased with age at a rate directly correlated to the molecular weight of aggregates forming the different chromatographic fractions, while tryptophan fluorescence tended to decrease. The crystallins constituting each fraction were separated using 2D-electrophoresis and after development with Coomassie blue they were identified using MS-TOF. Also, the protein content of each spot was quantified by subsequent scanning and integration. The proportions of unchanged crystallins characteristically changed with age in chromatographic fractions of different molecular weights. Thus it was possible to relate these changes with those occurring in the fluorescent properties and molecular weight of supramolecular structures.