Browsing by Author "Escribano, Damian"
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Item Changes in serum proteins in dogs with Ehrlichia canis infection(Elsevier, 2017-10-13) Escribano, Damian; Martinez, Silvia Subiela; Ceron, Jose Joaquin; Tvarijonaviciute, Asta; Cihan, Hüseyin; Levent, Pınar; Kocatürk, Meriç; Aytuğ, Nilüfer; Yılmaz, Zeki; Uludağ Üniversitesi/Veteriner Fakültesi/İç Hastalıkları Anabilim Dalı.; 0000-0001-9836-0749; ABH-3742-2020; V-5578-2017; 6602381681; 56690616700; 36437200800; 6505835923; 35944810500The aim of this study was the identification of proteins differentially represented in the serum proteome of seropositive dogs with (Group 1) and without (Group 2) clinical-pathologic signs consistent with ehrlichiosis compared to healthy control dogs. Serum samples were collected from 20 dogs of various breeds with naturally occurring ehrlichiosis (10 dogs belonged to Group 1 and 10 to Group 2) and 10 healthy dogs. Two-dimensional electrophoresis (2DE) of pooled serum for each of the group of dogs were run in triplicate. 2D image analysis showed 39 spots differently expressed between Group 1 and Group 2 compared with healthy ones. Mass spectrometry analysis allowed identification of 6 proteins: albumin, haptoglobin (Hp), alpha-l-antitrypsin (AAT), Retinol Binding Protein 4 (RBP-4), alpha-l-acid glycoprotein (AGP) and vitamin D-binding protein (VDBP). When a confirmatory study was performed for albumin, Hp, AAT and RBP-4 by using different assays, significant differences (P < 0.05) between diseased and healthy groups were observed. It can be concluded that there are significant changes in the serum proteome of dogs with ehrlichiosis with modifications in proteins related with the acute phase response such as Hp, albumin and AGP, with vitamin A transport such as RBP-4, with inhibitors of serine proteases and anti-inflammatory proteins such as AAT, and vitamin D metabolism and actin scavengers such as VDBP.Item Identification of novel biomarkers for treatment monitoring in canine leishmaniosis by high-resolution quantitative proteomic analysis(Elsevier, 2017-08-08) Martinez, Subiela Silvia; Horvatic, Anita; Escribano, Damian; Pardo, Luis Marin; Mrljak, Vladimir; Burchmore, Richard; Ceron, Jose J.; Kocatürk, Meriç; Yılmaz, Zeki; Uludağ Üniversitesi/Veteriner Fakültesi/İç Hastalıkları Anabilim Dalı.; 0000-0001-9836-0749; V-5578-2017; 36437200800; 35944810500The objective of this study was to use the Tandem Mass Tag (TMT) isobaric label-based proteomic approach, in order to identify new potential biomarkers for the treatment monitoring of canine leishmaniosis that could not be identified by the use of gel-based techniques. For this purpose serum samples were obtained from 5 clinically diseased dogs before and one month after the treatment of canine leishmaniosis. The non-depleted serum samples were subjected to reduction, alkylation and trypsin digestion, and the resulting peptides were labeled using 6-plex TMT reagents. To obtain information about protein identities and relative quantification, liquid chromatography-MS analysis of multiplexed TMT-labeled peptides was employed. This gel-free, label-based quantitative proteomic approach enabled identification of 117 canine proteins. Among these, 23 showed significant difference (p < 0.05) in expression (two downregulated and 21 upregulated ranging from 1.25 to 2.5 fold change). Comparison of gel-free TMT-based quantification and a gel-based approach previously applied to the same samples resulted in the identification of some common markers (Apo-A1, vitamin D binding protein and RBP4). However, 20 additional differentially represented proteins were highlighted by the gel-free approach, 13 of which have not been previously reported in canine leishmaniosis. In conclusion, the TMT-based proteomic approach allowed identification of new serum proteins that significantly change in concentration after canine leishmaniosis treatment. These proteins are involved in various physiopathological processes such as inflammatory, coagulation or defense mechanisms, and could potentially be suitable biomarkers for treatment monitoring of this parasitic disease.Item Serum apolipoprotein-A1 as a possible biomarker for monitoring treatment of canine leishmaniosis(Elsevier, 2016-10-10) Escribano, Damian; Tvarijonaviciute, Asta; Joaquin Ceron, Jose; Pardo-Marin, Luis; Torrecillas, Alejandro; Martinez-Subiela, Silvia; Kocatürk, Meriç; Yılmaz, Zeki; Uludağ Üniversitesi/Veteriner Fakültesi/Dahiliye Anabilim Dalı.; 0000-0001-9836-0749; V-5578-2017; 36437200800; 35944810500The aims of this study were: the identification of proteins differentially represented in the serum proteome of dogs with leishmaniosis after treatment and the verification of one selected protein as a possible biomarker for treatment monitoring. Serum samples from five dogs with leishmaniosis, before and after treatment were pooled into two groups and analysed using 2-dimensional electrophoresis followed by mass spectrometry analysis (MS). The MS analysis allowed the identification of 8 proteins differently expressed. APO-A1 was selected and an immunoturbidimetric assay was validated for its measurement in dogs. Significantly decreased concentrations of APO-A1 in dogs with leishmaniosis and a significant increase after a good response to the treatment were observed, suggesting that APO-A1 could be a potential biomarker of treatment monitoring with the advantages of an automated measurement.