Wound-Healing Markers Revealed by Proximity Extension Assay in Tears of Patients following Glaucoma Surgery

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Investigation of Neutral Losses and the Citrulline Effect for Modified H4 N-Terminal Pentapeptides

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A gluten-enteropathia kialakulását befolyásoló tényezők vizsgálata = Factors influencing the development of coeliac disease

Kutatásunk során új hajlamosító géneket írtunk le, melyet közrejátszhatnak a gluten-enteropathia (coeliakia) kialakulásában, ezek befolyásolják a T lymphocyták érését és kollaborációját a B sejtekkel, a toll-like receptorokat és a gyulladás során termelődő cytokineket. Megállapítottuk, hogy a coeliakia kialakulása és klinikai megjelenése asszociációt mutat a haptoglobin polimorfizmussal is, melyről a mi közlésünket követően független kutatók igazolták, hogy egyik altípusa azonos vékonybél permeabilitásában kulcsszerepet játszó zonulin molekulával. A kutatás során azonosítottuk a coeliakia-specifikus autoantitestek fő kötőhelyét az autoantigén, a 2-es típusú transzglutamináz enzim felszínén. Megállapítottuk, hogy a coeliakia antitestek betegség-specifikusan ugyanazt a komplex, háromdimenziós epitópot ismerik fel. Sejtkultúrában a coeliakia antitestek gátolják az erek képződését és fokozzák azok permeabilitását, elsősorban a RhoA túlzott aktiválásán keresztül. Megszerveztük és elvégeztük az első reprezentatív coeliakia szűrővizsgálatot Magyarországon. Megállapítottuk, hogy a coeliakia hazai előfordulása 1.3% körül van a magyar gyermekek és fiatal felnőttek körében. A szűrővizsgálatokra egyszerű, helyszíni antitest kimutatáson alapuló szűrő eljárást validáltunk. Kimutattuk, hogy a klinikailag fel nem ismert coeliakia kedvezőtlenül befolyásolja a hepatitis védőoltásra adott immunválaszt és az antitestek szerepet játszanak idegrendszeri szövődmény (ataxia) kialakulásában. | This research identified new susceptibility genes for coeliac disease, a common autoimmune enteropathy triggered by gluten peptides ingested with dietary cereals. The novel genes are mostly related to T-lymphocyte development, collaboration of T and B cells, toll-like receptors and cytokines. We also described association with haptoglobin polymorphism, a master regulator of intestinal permeability also known as zonulin. Predisposing alleles of these genes create a more inflammatory environment in the gut and in multiple copies confer additional risk to HLA-DQ alleles. We identified the main binding epitope of coeliac disease anti-transglutaminase 2 autoantibodies. This epitope has a complex, three-dimensional structure and is disease-specific. Patient antibodies targeting this epitope enhance the enzymatic activity of transglutaminase 2 and cause disturbances in the angiogenesis and vascular permeability in cell culture models via the inappropriate activation of RhoA. We performed the first representative population screening for coeliac disease in Hungary and established that the prevalence is around 1.3% in both children and young adults. The methodology of the screening has been advanced by the application of rapid, onsite antibody detection. Unrecognised coeliac disease was found to predispose to a defective immune response to hepatitis B vaccination (reversible after treatment) and we showed the contribution of coeliac disease antibodies in the late neural complications

Analysis of networks of host proteins in the early time points following HIV transduction

Background: Utilization of quantitative proteomics data on the network level is still a challenge in proteomics data analysis. Currently existing models use sophisticated, sometimes hard to implement analysis techniques. Our aim was to generate a relatively simple strategy for quantitative proteomics data analysis in order to utilize as much of the data generated in a proteomics experiment as possible. Results: In this study, we applied label-free proteomics, and generated a network model utilizing both qualitative, and quantitative data, in order to examine the early host response to Human Immunodeficiency Virus type 1 (HIV-1). A weighted network model was generated based on the amount of proteins measured by mass spectrometry, and analysis of weighted networks and functional sub-networks revealed upregulation of proteins involved in translation, transcription, and DNA condensation in the early phase of the viral life-cycle. Conclusion: A relatively simple strategy for network analysis was created and applied to examine the effect of HIV-1 on host cellular proteome. We believe that our model may prove beneficial in creating algorithms, allowing for both quantitative and qualitative studies of proteome change in various biological and pathological processes by quantitative mass spectrometry.Hungarian Scientific Research Fund [NKFI-6, 125238]; Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary [GINOP-2.3.3-15-2016-00020]; Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences; NIEHS [ES06694]; NIH/NCI [CA023074]; BIO5 Institute of the University of Arizona; NIH/NCRR [1S10 RR028868-01]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Wound-Healing Markers Revealed by Proximity Extension Assay in Tears of Patients following Glaucoma Surgery

Tears are a constantly available and highly valuable body fluid collectable by non-invasive techniques. Although it can give information on ocular status and be used for follow-ups, tear analysis is challenging due to the low amount of sample that is available. Proximity extension assay (PEA) allows for a sensitive and scalable analysis of multiple proteins in a single run from a one-mu L sample, so we applied this technique and examined the amount of 184 proteins in tears collected at different time points after trabeculectomy. The success rate of this surgical intervention highly depends on proper wound healing; therefore, information on the process is indispensable. We observed significantly higher levels of IL-6 and MMP1 at the early time points (day one, two, and four) following trabeculectomy, and the protein amounts went back to the level observed before the surgery three months after the intervention. Patients with or without complications were tested, and proteins that have roles in the immune response and wound healing could be observed with altered frequency and amounts in the cases of patients with complications. Our results highlight the importance of inflammation in wound-healing complications, and at the same time, indicate the utility of PEA in tear analysis

Diabetic retinopathy: Proteomic approaches to help the differential diagnosis and to understand the underlying molecular mechanisms

Diabetic retinopathy is the most common diabetic eye disease and a leading cause of blindness among patients with diabetes. The appearance and the severity of the symptoms correlate with the duration of diabetes and poor blood glucose level management. Diabetic retinopathy is also categorized as a chronic low-level inflammatory disease; the high blood glucose level promotes the accumulation of the advanced glycation end products and leads to the stimulation of monocytes and macrophages. Examination of protein level alterations in tears using state-of the art proteomics techniques have identified several proteins as possible biomarkers for the different stages of the diabetic retinopathy. Some of the differentially expressed tear proteins have a role in the barrier function of tears linking the diabetic retinopathy with another eye complication of diabetes, namely the diabetic keratopathy resulting in impaired wound healing. Understanding the molecular events leading to the eye complications caused by hyperglycemia may help the identification of novel biomarkers as well as therapeutic targets in order to improve quality of life of diabetic patients. BIOLOGICAL SIGNIFICANCE: Diabetic retinopathy (DR), the leading cause of blindness among diabetic patients can develop without any serious symptoms therefore the early detection is crucial. Because of the increasing prevalence there is a high need for improved screening methods able to diagnose DR as soon as possible. The non-invasive collection and the relatively high protein concentration make the tear fluid a good source for biomarker discovery helping the early diagnosis. In this work we have reviewed the administration of advanced proteomics techniques used in tear biomarker studies and the identified biomarkers with potential to improve the already existing screening methods for DR detection