Nasal secretory protein changes following intravenous choline administration in calves with experimentally induced endotoxaemia.

Nasal secretory fluid proteomes (NSPs) can provide valuable information about the physiopathology and prognosis of respiratory tract diseases. This study aimed to determine changes in NSP by using proteomics in calves treated with lipopolysaccharide (LPS) or LPS + choline. Healthy calves (n = 10) were treated with LPS (2 µg/kg/iv). Five minutes after LPS injection, the calves received a second iv injection with saline (n = 5, LPS + saline group) or saline containing 1 mg/kg choline (n = 5, LPS + choline group). Nasal secretions were collected before (baseline), at 1 h and 24 h after the treatments and analysed using label-free liquid chromatography-tandem mass spectrometry (LCMS/MS). Differentially expressed proteins (>1.2-fold-change) were identified at the different time points in each group. A total of 52 proteins were up- and 46 were downregulated at 1 h and 24 h in the LPS + saline group. The upregulated proteins that showed the highest changes after LPS administration were small ubiquitin-related modifier-3 (SUMO3) and glutathione peroxidase-1 (GPX1), whereas the most downregulated protein was E3 ubiquitin-protein ligase (TRIM17). Treatment with choline reduced the number of upregulated (32 proteins) and downregulated proteins (33 proteins) in the NSPs induced by LPS. It can be concluded that the proteome composition of nasal fluid in calves changes after LPS, reflecting different pathways, such as the activation of the immunological response, oxidative stress, ubiquitin pathway, and SUMOylation. Choline treatment alters the NSP response to LPS.

Identification of candidate biomarkers in converting and non-converting clinically isolated syndrome by proteomics analysis of cerebrospinal fluid.

Multiple sclerosis (MS) often starts in the form of clinically isolated syndrome (CIS) and only some of the CIS patients progress to relapsing-remitting MS (RRMS). Biomarkers to predict conversion from CIS to MS are thus greatly needed for making correct treatment decisions. To identify a predictive cerebrospinal fluid (CSF) protein, we analyzed the first-attack CSF samples of CIS patients who converted (CIS-MS) (n = 23) and did not convert (CIS-CIS) (n = 19) to RRMS in a follow-up period of 5 years using proteomics analysis by liquid chromatography tandem-mass spectrometry (LC-MS/MS) and verified by ELISA. Label-free differential proteomics analysis of CSF ensured that 637 proteins were identified and 132 of these proteins were found to be statistically significant. Further investigation with the ingenuity pathway analysis (IPA) software led to identification of three pathway networks mostly comprised proteins involved in inflammatory response, cellular growth and tissue proliferation. CSF levels of four of the most differentially expressed proteins belonging to the cellular proliferation network function, chitinase-3-like protein 1 (CHI3L1), tumor necrosis factor receptor superfamily member 21 (TNFRSF21), homeobox protein Hox-B3 (HOXB3) and iduronate 2-sulfatase (IDS), were measured by ELISA. CSF levels of HOXB3 were significantly increased in CIS-MS patients. Our results indicate that cell and tissue proliferation functions are dysregulated in MS as early as the first clinical episode. HOXB3 has emerged as a potential novel biomarker which might be used for prediction of CIS-MS conversion.

Changes in serum proteins after endotoxin administration in healthy and choline-treated calves.

BACKGROUND: This study aimed to investigate the possible serum protein changes after endotoxin administration in healthy and choline-treated calves using proteomics. These results are expected to contribute to the understanding of the pathophysiological mechanisms of endotoxemia and the beneficial effect of choline administration in this clinical situation. METHODS: Healthy-calves (n = 20) were divided into 4 groups: Control, Choline treated (C), Lipopolysaccharide administered (LPS), and LPS + C. Control calves received 0.9 % NaCl injection. Calves in C and LPS + C groups received choline chloride (1 mg/kg/iv). Endotoxin (LPS) was injected (2 µg/kg/iv) to the calves in LPS and LPS + C groups. Serum samples were collected before and after the treatments. Differentially expressed proteins (> 1.5 fold-change relative to controls) were identified by LC-MS/MS. RESULTS: After LPS administration, 14 proteins increased, and 13 proteins decreased within 48 h as compared to controls. In the LPS group, there were significant increases in serum levels of ragulator complex protein (189-fold) and galectin-3-binding protein (10-fold), but transcription factor MafF and corticosteroid binding globulin were down regulated (≥ 5 fold). As compared with the LPS group, in LPS + C group, fibrinogen gamma-B-chain and antithrombin were up-regulated, while hemopexin and histone H4 were down-regulated. Choline treatment attenuated actin alpha cardiac muscle-1 overexpression after LPS. CONCLUSIONS: LPS administration produces changes in serum proteins associated with lipid metabolism, immune and inflammatory response, protein binding/transport, cell adhesion, venous thrombosis, cardiac contractility and blood coagulation. The administration of choline is associated with changes in proteins which can be related with its beneficial effect in this clinical situation.