Investigation of the Abundance of Proteins Secreted by Fasciola hepatica, Which is Exposed to Environmental Change in Experimental Studies, with an Advanced Proteomic Approach

Haçarız, Orçun; Baykal, Ahmet Tarık

doi:10.5152/tpd.2014.3443

Cited by 4 publications

(4 citation statements)

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“…In order to analyse differences in protein expression in the course of the study, three serum samples at each time point for each treatment were obtained and the protein profiles of the serum samples were analysed by LC-MS/MS method [18][19][20]. Albumin depletion from serum samples was done with albumin depletion spin columns (Pierce, Thermo Scientific).…”

Section: Serum Proteomic Analysismentioning

confidence: 99%

“…IgGs from the albumin-depleted serum samples were also depleted with an Albumin/IgG Removal Kit (Pierce, Thermo scientific). The remaining serum proteins (at same concentration for all samples) were extracted and enzymatically digested into peptides, and the resulting peptides were subjected to the nanoUPLC-ESI-qTOF-MS system (nanoultra-performance LC and ESI quadrupole TOF MS) (Waters) with the application of data independent acquisition method also known as the MS E as previously described in detail [20]. Minimum 3 technical replicates for each sample were used for the analysis.…”

Section: Serum Proteomic Analysismentioning

confidence: 99%

“…The proteomic data was searched against a reviewed database [including protein sequences of Bos taurus and an internal standard (enolase, Saccharomyces cerevisiae; #P00924), obtained from UniProt (htpp://www.uniprot.org)] with previously described in silico parameters [19]. Quantitative differences of proteins at different time points were calculated using Progenesis LC-MS software V4.0 (Nonlinear Dynamics) [20].…”

Section: Serum Proteomic Analysismentioning

confidence: 99%

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

et al. 2016

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BackgroundThis 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.MethodsHealthy-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.ResultsAfter 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.ConclusionsLPS 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-016-0837-y) contains supplementary material, which is available to authorized users.

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Section: Serum Proteomic Analysismentioning

confidence: 99%

Section: Serum Proteomic Analysismentioning

confidence: 99%

Section: Serum Proteomic Analysismentioning

confidence: 99%

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

et al. 2016

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“…Adult liver flukes were collected from the bile ducts of naturally infected cattle in an abottoir (Tuzla, İstanbul) and immediately placed in a 50 ml tube containing warm PBS (Biochrom) as previously described [ 74 ]. Intact liver flukes were washed with warm PBS and placed in a flask containing culture medium (DMEM, Invitrogen) and gentamycin (50 μg as a final concentration) (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

Comparative transcriptome profiling approach to glean virulence and immunomodulation-related genes of Fasciola hepatica

et al. 2015

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BackgroundFasciola hepatica causes chronic liver disease, fasciolosis, leading to significant losses in the livestock economy and concerns for human health in many countries. The identification of F. hepatica genes involved in the parasite’s virulence through modulation of host immune system is utmost important to comprehend evasion mechanisms of the parasite and develop more effective strategies against fasciolosis. In this study, to identify the parasite’s putative virulence genes which are associated with host immunomodulation, we explored whole transcriptome of an adult F. hepatica using current transcriptome profiling approaches integrated with detailed in silico analyses. In brief, the comparison of the parasite transcripts with the specialised public databases containing sequence data of non-parasitic organisms (Dugesiidae species and Caenorhabditis elegans) or of numerous pathogens and investigation of the sequences in terms of nucleotide evolution (directional selection) and cytokine signaling relation were conducted.ResultsNGS of the whole transcriptome resulted in 19,534,766 sequence reads, yielding a total of 40,260 transcripts (N50 = 522 bp). A number of the parasite transcripts (n = 1,671) were predicted to be virulence-related on the basis of the exclusive homology with the pathogen-associated data, positive selection or relationship with cytokine signaling. Of these, a group of the virulence-related genes (n = 62), not previously described, were found likely to be associated with immunomodulation based on in silico functional categorisation, showing significant sequence similarities with various immune receptors (i.e. MHC I class, TGF-β receptor, toll/interleukin-1 receptor, T-cell receptor, TNF receptor, and IL-18 receptor accessory protein), cytokines (i.e. TGF-β, interleukin-4/interleukin-13 and TNF-α), cluster of differentiations (e.g. CD48 and CD147) or molecules associated with other immunomodulatory mechanisms (such as regulation of macrophage activation). Some of the genes (n = 5) appeared to be under positive selection (Ka/Ks > 1), imitating proteins associated with cytokine signaling (through sequence homologies with thrombospondin type 1, toll/interleukin-1 receptor, TGF-β receptor and CD147).ConclusionsWith a comparative transcriptome profiling approach, we have identified a number of potential immunomodulator genes of F. hepatica (n = 62), which are firstly described here, could be employed for the development of better strategies (including RNAi) in the battle against both zoonotically and economically important disease, fasciolosis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1539-8) contains supplementary material, which is available to authorized users.

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The omic approach to parasitic trematode research—a review of techniques and developments within the past 5 years

Haçarız

Sayers

2016

Parasitol Res

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The evolution of technologies to explore parasite biology at a detailed level has made significant advances in recent years, particularly with the development of omic-based strategies. Whilst extensive efforts have been made in the past to develop therapeutic and prophylactic control strategies for trematode parasites, only the therapeutic anthelmintic approach can be regarded as usable in clinical practice. Currently, there is no commercialised prophylactic strategy (such as vaccination) for protection of the definitive host against any trematode parasite. Since 2010 in particular, the integration of omic technologies, including liquid chromatography-mass spectrometry (LC-MS) and next-generation sequencing (NGS), has been increasingly reported in trematode-related studies. Both LC-MS and NGS facilitate a better understanding of the biology of trematodes and provide a promising route to identifying clinically important biological characteristics of parasitic trematodes. In this review, we focus on the application, advantages, and disadvantages of omic technologies (LC-MS and NGS) in trematode research within the past 5 years and explore the use and translation of the omic-based research results into practical tools to deal with infection.

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Investigation of the Abundance of Proteins Secreted by Fasciola hepatica, Which is Exposed to Environmental Change in Experimental Studies, with an Advanced Proteomic Approach

Cited by 4 publications

References 14 publications

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

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

Comparative transcriptome profiling approach to glean virulence and immunomodulation-related genes of Fasciola hepatica

The omic approach to parasitic trematode research—a review of techniques and developments within the past 5 years

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