Institute For Research And Technology Thessaly Ireteth
Institute For Research And Technology Thessaly Ireteth
Melagraki G.,Biomedical science Research Center Alexander Fleming |
Melagraki G.,NovaMechanics Ltd |
Ntougkos E.,Biomedical science Research Center Alexander Fleming |
Rinotas V.,Biomedical science Research Center Alexander Fleming |
And 14 more authors.
PLoS Computational Biology | Year: 2017
We present an in silico drug discovery pipeline developed and applied for the identification and virtual screening of small-molecule Protein-Protein Interaction (PPI) compounds that act as dual inhibitors of TNF and RANKL through the trimerization interface. The cheminformatics part of the pipeline was developed by combining structure–based with ligand–based modeling using the largest available set of known TNF inhibitors in the literature (2481 small molecules). To facilitate virtual screening, the consensus predictive model was made freely available at: http://enalos.insilicotox.com/TNFPubChem/. We thus generated a priority list of nine small molecules as candidates for direct TNF function inhibition. In vitro evaluation of these compounds led to the selection of two small molecules that act as potent direct inhibitors of TNF function, with IC50values comparable to those of a previously-described direct inhibitor (SPD304), but with significantly reduced toxicity. These molecules were also identified as RANKL inhibitors and validated in vitro with respect to this second functionality. Direct binding of the two compounds was confirmed both for TNF and RANKL, as well as their ability to inhibit the biologically-active trimer forms. Molecular dynamics calculations were also carried out for the two small molecules in each protein to offer additional insight into the interactions that govern TNF and RANKL complex formation. To our knowledge, these compounds, namely T8 and T23, constitute the second and third published examples of dual small-molecule direct function inhibitors of TNF and RANKL, and could serve as lead compounds for the development of novel treatments for inflammatory and autoimmune diseases. © 2017 Melagraki et al.
Mavropoulos A.,Institute For Research And Technology Thessaly Ireteth |
Mavropoulos A.,King's College London |
Orfanidou T.,King's College London |
Liaskos C.,King's College London |
And 7 more authors.
Autoimmune Diseases | Year: 2013
p38 mitogen activated protein kinase (p38 MAPK) signaling plays a major role in the modulation of immune-mediated inflammatory responses and therefore has been linked with several autoimmune diseases. The extent of the involvement of p38 MAPK in the pathogenesis of autoimmune blistering diseases has started to emerge, but whether it pays a critical role is a matter of debate. The activity of p38 MAPK has been studied in great detail during the loss of keratinocyte cell-cell adhesions and the development of pemphigus vulgaris (PV) and pemphigus foliaceus (PF). These diseases are characterised by autoantibodies targeting desmogleins (Dsg). Whether autoantibody-antigen interactions can trigger signaling pathways (such as p38 MAPK) that are tightly linked to the secretion of inflammatory mediators which may perpetuate inflammation and tissue damage in pemphigus remains unclear. Yet, the ability of p38 MAPK inhibitors to block activation of the proapoptotic proteinase caspase-3 suggests that the induction of apoptosis may be a consequence of p38 MAPK activation during acantholysis in PV. This review discusses the current evidence for the role of p38 MAPK in the pathogenesis of pemphigus. We will also present data relating to the targeting of these cascades as a means of therapeutic intervention. © 2013 Athanasios Mavropoulos et al.
Liaskos C.,Institute for Research and Technology Thessaly IReTeTh |
Liaskos C.,University of Thessaly |
Spyrou V.,Technological Educational Institute of Larissa |
Athanasiou L.V.,University of Thessaly |
And 9 more authors.
Clinics and Research in Hepatology and Gastroenterology | Year: 2015
Background: Pancreatic autoantibodies (PABs) specifically recognizing GP2 and/or CUZD1 are present in more than 35% of patients with Crohn's disease (CrD). We have recently provided evidence of the presence of GP2-specific PABs in ruminants with paratuberculosis (ptb), a Mycobacterium avium paratuberculosis (MAP)-induced disease resembling CrD. Objective: To assess whether anti-CUZD1 antibodies are also present in ruminants with ptb. Methods: A total of 110 samples (73 cattle/37 sheep) were studied including 40 with ptb (24 cattle/16 sheep; 20 anti-GP2 antibody pos) and 70 without ptb (49 cattle/21 sheep; 10 anti-GP2 antibody pos). The samples were pre-characterized for anti-MAP and anti-GP2 antibodies by ELISA. Evidence of MAP was confirmed by PCR. Anti-CUZD1 antibody testing was performed by indirect immunofluorescence (IIF) based on transfected HEK293 cells expressing CUZD1. Anti-sheep or anti-cattle specific antisera were used as revealing antibodies. Results: None of the ruminant sera had anti-CUZD1 antibodies by IIF testing at dilutions varying from 1/10 to 1/160. Methodological flaws were prevented by a series of tests. Control sera from anti-CUZD1 positive CrD samples have shown anti-CUZD1 antibody reactivity at various concentrations. Antibody reactivity to GP2-expressing HEK293 cells has confirmed the reactivity to GP2 in ruminant sera found positive for anti-GP2 antibodies by ELISA. Conclusion: The present study has found no evidence of anti-CUZD1 PABs in MAP-induced ptb. Our findings indicate that the induction of CUZD1-specific PABs is unrelated to MAP infection and that the mechanisms responsible for the loss of tolerance to GP2 and CUZD1 are probably quite distinct. © 2015 Elsevier Masson SAS.
Manthou E.,Technological Educational Institute of Larissa |
Manthou E.,University of Thessaly |
Manthou E.,Institute For Research And Technology Thessaly Ireteth |
Kanaki M.,Technological Educational Institute of Larissa |
And 10 more authors.
Nutrients | Year: 2014
In this study we examined the glycaemic index (GI) and glycaemic load (GL) of a functional food product, which contains ewe-goat whey protein and carbohydrates in a 1:1 ratio. Nine healthy volunteers, (age, 23.3 ± 3.9 years; body mass index, 24.2 ± 4.1 kg·m2; body fat %, 18.6 ± 10.0) randomly consumed either a reference food or amount of the test food both with equal carbohydrate content in two visits. In each visit, seven blood samples were collected; the first sample after an overnight fast and the remaining six at 15, 30, 45, 60, 90 and 120 min after the beginning of food consumption. Plasma glucose concentration was measured and the GI was determined by calculation of the incremental area under the curve. The GL was calculated using the equation: test food GI/100 g available carbohydrates per test food serving. The GI of the test food was found to be 5.18 ± 3.27, while the GL of one test food serving was 1.09 ± 0.68. These results indicate that the tested product can be classified as a low GI (<55) and low GL (<10) food. Given the health benefits of low glycaemic response foods and whey protein consumption, the tested food could potentially promote health beyond basic nutrition. © 2014 by the authors; licensee MDPI, Basel, Switzerland.
Koureas M.,University of Thessaly |
Tsezou A.,University of Thessaly |
Tsakalof A.,University of Thessaly |
Orfanidou T.,Institute For Research And Technology Thessaly Ireteth |
Hadjichristodoulou C.,University of Thessaly
Science of the Total Environment | Year: 2014
The widespread use of pesticides substances nowadays largely guarantees the protection of crops and people from undesired pests. However, exposure to pesticides was related to a variety of human health effects. The present study was conducted in the region of Thessaly which is characterized by intensive agricultural activities and wide use of pesticides. The study aimed at estimating the oxidative damage to DNA in different subpopulations in Thessaly region (Greece) and investigating its correlation with exposure to pesticides and other potential risk factors. In total, the study involved 80 pesticide sprayers, 85 rural residents and 121 individuals, inhabitants of the city of Larissa. Demographic characteristics, habits, medical history and exposure history of the participants to pesticides were recorded by personal interviews. Blood and urine samples were collected from all participants. For the measurement of exposure to organophosphorus insecticides, dialkylphosphate (DAP) metabolites were quantified in urine, by gas chromatography-mass spectrometry. Genomic DNA was extracted from peripheral blood samples and the oxidation by-product 8-hydroxydeoxyguanosine (8-OHdG) was determined by Enzyme Immuno-Assay. Urinary metabolite concentrations were not associated with 8-OHdG levels but it was found that pesticide sprayers had significantly higher levels of 8-OHdG (p= 0.007) in comparison to the control group. Last season's exposure to insecticides and fungicides, expressed as total area treated multiplied by the number of applications, showed a statistically significant association with the risk of having high 8-OHdG levels [RR: 2.19 (95%CI:1.09-4.38) and RR: 2.32 (95% CI:1.16-4.64) respectively]. Additionally, from the subgroups of pesticides examined, seasonal exposure to neonicotinoid insecticides [RR: 2.22 (95% CI:1.07-4.63)] and glufosinate ammonium [RR: 3.26 (95% CI:1.38-7.69)] was found to have the greater impact on 8-OHdG levels. This study produced findings that support the hypothesis that pesticide exposure is involved in the induction of oxidative damage to DNA and identified chemical groups of pesticides which should be given greater attention in future investigations. © 2014 Elsevier B.V.
Gkretsi V.,Institute For Research And Technology Thessaly Ireteth |
Papanikolaou V.,Institute For Research And Technology Thessaly Ireteth |
Zacharia L.C.,Institute For Research And Technology Thessaly Ireteth |
Athanassiou E.,University of Thessaly |
And 4 more authors.
Anticancer Research | Year: 2013
Background: Cell adhesion proteins that connect each cell to neighboring cells and the extracellular matrix play a fundamental role in metastasis. Mitogen-inducible gene-2 (MIG2), is a cell-matrix adhesion protein, which through migfilin, interacts with filamin-A, being linked to actin cytoskeleton. Aim: Recent studies have implicated both MIG2 and migfilin in cancer, but little is known regarding their expression in breast cancer. In this study, we investigated this topic. Materials and Methods: mRNA and protein expression was examined in 30 breast cancer samples and compared to that of normal adjacent tissue using real time-polymerase chain reaction (PCR) and western blotting. Results: Our results showed that expression of MIG2 and migfilin was significantly reduced in the majority of the breast cancer tissues compared to normal tissues regardless of metastatic status and disease stage. Conclusion: Both MIG2 and migfilin are down-regulated in breast cancer.
Lyberopoulou A.,University of Thessaly |
Lyberopoulou A.,Institute for Research and Technology Thessaly IRETETH |
Chachami G.,University of Thessaly |
Chachami G.,Institute for Research and Technology Thessaly IRETETH |
And 12 more authors.
PLoS ONE | Year: 2015
Hepcidin, a liver hormone, is important for both innate immunity and iron metabolism regulation. As dysfunction of the hepcidin pathway may contribute to liver pathology, we analysed liver hepcidin mRNA and serum hepcidin in patients with chronic liver diseases. Hepcidin mRNA levels were determined in liver biopsies obtained from 126 patients with HCV (n = 21), HBV (n = 23), autoimmune cholestatic disease (primary biliary cirrhosis and primary sclerosing cholangitis; PBC/PSC; n = 34), autoimmune hepatitis (AIH; n = 16) and non-alcoholic fatty liver disease (NAFLD; n = 32). Sera sampled on the biopsy day from the same patients were investigated for serum hepcidin levels. Hepatic hepcidin mRNA levels correlated positively with ferritin and negatively with serum γ-GT levels. However, no correlation was found between serum hepcidin and either ferritin or liver hepcidin mRNA. Both serum hepcidin and the serum hepcidin/ferritin ratio were significantly lower in AIH and PBC/PSC patients' sera compared to HBV, HCV or NAFLD (P< 0.001 for each comparison) and correlated negatively with serum ALP levels. PBC/PSC and AIH patients maintained low serum hepcidin during the course of their two-year long treatment. In summary, parallel determination of liver hepcidin mRNA and serum hepcidin in patients with chronic liver diseases shows that circulating hepcidin and its respective ratio to ferritin are significantly diminished in patients with autoimmune liver diseases. These novel findings, once confirmed by followup studies involving bigger size and better-matched disease subgroups, should be taken into consideration during diagnosis and treatment of autoimmune liver diseases. Copyright: © 2015 Lyberopoulou et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.