9 research outputs found
Hledání potenciálního vazebného partnera glutamátkarboxypeptidasy II pomocí hmotnostní spektrometrie
Český abstrakt Systémová biologie bude vyžadovat použití nových metod s vysokou propustností dat pro studium protein-proteinových interakcí. Proteomika založená na hmotnostní spektrometrii prošla v posledních letech rychlým vývojem a stala se nepostradatelným nástrojem studia makromolekul v základním i klinickém výzkumu. Glutamátkarboxypeptidasa II (GCPII; EC 3.5.17.21) je transmembránový protein se dvěma známými enzymovými aktivitami. Tento enzym je exprimován ve větším množství v buňkách některých pevných nádorů a také obecně v neovaskulatuře pevných nádorů, nehraje zde však fyziologickou roli. Některé skutečnosti mohou poukazovat na potenciální receptorovou funkci GCPII, přirozený ligand tohoto proteinu však nebyl doposud identifikován. Pro hledání možných vazebných partnerů a/nebo ligandů GCPII byly použity metody afinitní chromatografie, imunoprecipitace a hmotnostní spektrometrie. Byla optimalizována metodika přípravy vzorků pro hmotnostní spektrometrii a analýza získaných dat s cílem nalézt možné vazebné partnery. Jeden z identifikovaných proteinů- β-podjednotka F1 ATP-syntasy, byl vybrán pro důkladnější analýzu možné interakce s GCPII.English Abstract The incoming paradigm of the network (or systems) biology calls for a new high throughput tool for a wide scale study of protein-protein interactions. Mass spectrometry-based proteomics have experienced a great progress in recent years and have become an indispensable technology of elementary as well as clinical research. Glutamate carboxypeptidase II (GCPII; EC 3.5.17.21) is a transmembrane protein with two known enzymatic activities. Its expression is highly upregulated in some solid tumors and also in tumor-associated neovasculature in general. Nevertheless, none of the two enzymatic activities were shown to be physiologically relevant to these cells. Some facts point at a possible receptor function of GCPII, however, no specific binding partner has been found yet. In the search for potential binding partners and/or ligands of GCPII, a series of methods have been employed, including pull-down experiment, immunoprecipitation and mass spectrometry. Sample preparation and mass spectrometry data processing methodology was specifically developed in order to identify potential binding partners. As one of the outcome of that methodology, the interaction of β-subunit of F1 ATP synthase was selected for further detailed analysis as a putative ligand of GCPII.Department of BiochemistryKatedra biochemieFaculty of SciencePřírodovědecká fakult
Příprava a charakterisace rekombinantního dermcidinu jako potenciálního proteinového partnera glutamátkarboxypeptidasy II
A process of forming new blood vessels is necessary for tumour viability and expansion. Without vasculature, tumour stops growing at a size of millimeters. Some tumours, however, undergo an angiogenic switch and start to build up their own vascular architecture. The rate of apoptosis then decreases and the tumour becomes invasive. There are many factors that control the process of physiological angiogenesis. These might or might not relate to tumour tissue as well. Glutamate carboxypeptidase II (GCPII; EC 3.4.17.21) is a type II transmembrane glycoprotein with two known enzymatic activities. GCPII expression is upregulated in prostate cancer and also highly expressed in tumour-associated neovasculature even though none of these enzymatic functions was observed on the endothelium. Although numerous researches suggested that GCPII might serve as a receptor, no natural ligand has been identified yet. Preliminary experiments performed in our laboratory indicated some proteins to be possible natural ligands of GCPII. Therefore, we chose one of them- dermcidin, cloned and expressed this protein in mammalian cells. We investigated its possible interaction with GCPII introducing new detection system utilizing FLAG-tag however, we were not able to approve neither disapprove its interaction in vitro
Reconstruction of roads in the southern part of the settlement Višňovka in Pardubice
Rekonstrukce komunikací a veřejných ploch v dané oblasti. Komunikace jsou navrženy jako zóna tempo 30. Součástí návrhu je i navržení cyklistických pruhů, chodníků a bezbariérových prvků. Práce obsahuje směrové a výškové řešení, návrh parkovacích stání a napojení na přilehlé nemovitosti.Reconstruction ot roads and public areas of the region. Roads are designed as zone 30 tempo. Part of the propsal is the design of bicycle lanes, sidewalks and barrier free features. The work includes horizontál and vertical alignment, design of parking and connections to adjacent propertyKatedra dopravního stavitelstvíUchazeč prezentoval komisi hlavní výsledky a závěry své diplomové práce. Po prezentaci diplomové práce zodpověděl dotazy vedoucího a oponenta diplomové práce. V diskusi o diplomové práci podrobně zodpověděl všechny dotazy členů komise
Mass Spectrometry-Based Identification of a Potential Binding Partner of Glutamate Carboxypetidase II
English Abstract The incoming paradigm of the network (or systems) biology calls for a new high throughput tool for a wide scale study of protein-protein interactions. Mass spectrometry-based proteomics have experienced a great progress in recent years and have become an indispensable technology of elementary as well as clinical research. Glutamate carboxypeptidase II (GCPII; EC 3.5.17.21) is a transmembrane protein with two known enzymatic activities. Its expression is highly upregulated in some solid tumors and also in tumor-associated neovasculature in general. Nevertheless, none of the two enzymatic activities were shown to be physiologically relevant to these cells. Some facts point at a possible receptor function of GCPII, however, no specific binding partner has been found yet. In the search for potential binding partners and/or ligands of GCPII, a series of methods have been employed, including pull-down experiment, immunoprecipitation and mass spectrometry. Sample preparation and mass spectrometry data processing methodology was specifically developed in order to identify potential binding partners. As one of the outcome of that methodology, the interaction of β-subunit of F1 ATP synthase was selected for further detailed analysis as a putative ligand of GCPII
Design of the Stěžery by-pass road
Jedná se o návrh obchvatu obce Stěžery. Bylo navrženo směrové a výškové vedení komunikace, včetně konstrukce vozovky a odvodnění. Obchvat je navržen jako kategorie S9,5/80.The work is the proposal shifting of the road II/324 in Stěžery. It was designed directional and height lead of communication include pavement construction and drainage. Bypass is designed like category S9,5/80.Katedra dopravního stavitelstvíUchazeč prezentoval komisi hlavní výsledky a závěry své bakalářské práce. Po prezentaci bakalářské práce zodpověděl dotazy vedoucího a oponenta bakalářské práce. V diskusi o bakalářské práci podrobně zodpověděl všechny dotazy členů komise.Dokončená práce s úspěšnou obhajobo
Příprava a charakterisace rekombinantního dermcidinu jako potenciálního proteinového partnera glutamátkarboxypeptidasy II
A process of forming new blood vessels is necessary for tumour viability and expansion. Without vasculature, tumour stops growing at a size of millimeters. Some tumours, however, undergo an angiogenic switch and start to build up their own vascular architecture. The rate of apoptosis then decreases and the tumour becomes invasive. There are many factors that control the process of physiological angiogenesis. These might or might not relate to tumour tissue as well. Glutamate carboxypeptidase II (GCPII; EC 3.4.17.21) is a type II transmembrane glycoprotein with two known enzymatic activities. GCPII expression is upregulated in prostate cancer and also highly expressed in tumour-associated neovasculature even though none of these enzymatic functions was observed on the endothelium. Although numerous researches suggested that GCPII might serve as a receptor, no natural ligand has been identified yet. Preliminary experiments performed in our laboratory indicated some proteins to be possible natural ligands of GCPII. Therefore, we chose one of them- dermcidin, cloned and expressed this protein in mammalian cells. We investigated its possible interaction with GCPII introducing new detection system utilizing FLAG-tag however, we were not able to approve neither disapprove its interaction in vitro
Mass Spectrometry-Based Identification of a Potential Binding Partner of Glutamate Carboxypetidase II
English Abstract The incoming paradigm of the network (or systems) biology calls for a new high throughput tool for a wide scale study of protein-protein interactions. Mass spectrometry-based proteomics have experienced a great progress in recent years and have become an indispensable technology of elementary as well as clinical research. Glutamate carboxypeptidase II (GCPII; EC 3.5.17.21) is a transmembrane protein with two known enzymatic activities. Its expression is highly upregulated in some solid tumors and also in tumor-associated neovasculature in general. Nevertheless, none of the two enzymatic activities were shown to be physiologically relevant to these cells. Some facts point at a possible receptor function of GCPII, however, no specific binding partner has been found yet. In the search for potential binding partners and/or ligands of GCPII, a series of methods have been employed, including pull-down experiment, immunoprecipitation and mass spectrometry. Sample preparation and mass spectrometry data processing methodology was specifically developed in order to identify potential binding partners. As one of the outcome of that methodology, the interaction of β-subunit of F1 ATP synthase was selected for further detailed analysis as a putative ligand of GCPII
Relationship between Patient Preferences, Attitudes to Treatment, Adherence, and Quality of Life in New Users of Teriflunomide
Background: A poor patient adherence often limits the real-world effectiveness of an oral disease-modifying therapy (DMT) for multiple sclerosis (MS). In the present study, we aimed to map patient preferences, attitudes toward treatment, and quality of life to identify the predictors of non-adherence to teriflunomide. Methods: This was a single-arm, non-interventional, multicenter study (Czech Act 378/2007 Coll.) consisting of three visits: the first at treatment initiation (teriflunomide 14 mg), and then after 3 and 9 months of therapy. We enrolled both DMT-naïve and patients who had undergone a DMT diagnosed with a clinically isolated syndrome (CIS) or relapsing-remitting multiple sclerosis (RRMS). The functional status and MS activity were estimated using the Expanded Disability Status Scale (EDSS) and annualized relapse rate (ARR); the quality of life via the Multiple Sclerosis Impact Scale (MSIS-29); the medication adherence with the Morisky Medication Adherence Scale (MMAS-8); the confidence in the ability to take medications by the Self-Efficacy for Appropriate Medication Score (SEAMS); and the attitude to the therapy via the Beliefs about Medicines Questionnaire (BMQ). After nine months of therapy, we predicted the adherence to teriflunomide (MMAS-8) by fitting a multivariate ordinal logistic model with EDSS changes, gender, previous DMT, MSIS-29, BMQ, and SEAMS as the explanatory variables. Results: Between 2018 and 2019, 114 patients were enrolled at 10 sites in the Czech Republic. The mean age was 41.2 years, 64.8% were diagnosed with a CIS, 52.4% were DMT-naïve, and 98.1% of patients preferred an oral administration at the baseline. The mean EDSS baseline was 1.97 and remained constant during the 9 months of therapy. The ARR baseline was 0.72 and dropped to 0.19 and 0.15 after 3 and 9 months, respectively. Despite a more than 4-fold higher ARR baseline, the treatment-naïve patients achieved an ARR at 9 months comparable with those previously treated. There were ten non-serious adverse reactions. After nine months of teriflunomide therapy, 63.3%, 21.2%, and 15.4% of patients had a high, medium, and low adherence, respectively, as per the MMAS-8; 100% of patients preferred an oral administration. The SEAMS score (odds ratio (OR) = 0.91; p = 0.013) and previous DMT (OR = 4.28; p = 0.005) were the only significant predictors of non-adherence. The disability, the quality of life, and beliefs about medicines had no measurable effect on adherence. Conclusion: After nine months of teriflunomide therapy, both the disability and quality of life remained stable; the relapse rate significantly decreased, 63.3% of patients had a high adherence, and 100% of patients preferred an oral administration. A low adherence was associated with previous DMT experiences and a low self-efficacy for the appropriate medication (i.e., the confidence in one’s ability to take medication correctly)
Místo vstupu neovlivňuje riziko cévní mozkové příhody po diagnostické koronární agniografii nebo intervenci: výsledky z velkého prospektivního registru
Introduction: Periprocedural stroke represents a rare but serious complication of cardiac catheterization. Pooled data from randomized trials evaluating the risk of stroke following cardiac catheterization via transradial versus transfemoral access showed no difference. On the other hand, a significant difference in stroke rates favoring transradial access was found in a recent meta-analysis of observational studies. Our aim was to determine if there is a difference in stroke risk after transradial versus transfemoral catheterization within a contemporary real-world registry. Methods: Data from 14,139 patients included in a single-center prospective registry between 2009 and 2016 were used to determine the odds of periprocedural transient ischemic attack (TIA) and stroke for radial versus femoral catheterization via multivariate logistic regression with Firth's correction. Results: A total of 10,931 patients underwent transradial and 3,208 underwent transfemoral catheterization. Periprocedural TIA/stroke occurred in 41 (0.29%) patients. Age was the only significant predictor of TIA/stroke in multivariate analysis, with each additional year representing an odds ratio (OR) = 1.09 (CI 1.05-1.13, p < 0.000). The choice of accession site had no impact on the risk of periprocedural TIA/stroke (OR = 0.81; CI 0.38-1.72, p = 0.577). Conclusion: Observational data from a large prospective registry indicate that accession site has no influence on the risk of periprocedural TIA/stroke after cardiac catheterization. Keywords: Cardiac catheterization; Femoral access; Radial access; Stroke; Transient ischemic attack.Úvod: Periprocedurální cévní mozková příhoda představuje vzácnou, ale závažnou komplikaci srdeční katetrizace. Shromážděné údaje z randomizovaných studií hodnotících riziko cévní mozkové příhody po srdeční katetrizaci transradiálním versus transfemorálním přístupem neukázaly žádný rozdíl. Na druhé straně byl v nedávné metaanalýze observačních studií zjištěn významný rozdíl v četnosti cévních mozkových příhod ve prospěch transradiálního přístupu. Naším cílem bylo zjistit, zda existuje rozdíl v riziku cévní mozkové příhody po transradiální a transfemorální katetrizaci v rámci současného registru v reálném světě. Metodika: Data od 14 139 pacientů zařazených do jednocentrového prospektivního registru v letech 2009 až 2016 byla použita ke stanovení pravděpodobnosti periprocedurální tranzitorní ischemické ataky (TIA) a cévní mozkové příhody pro radiální versus femorální katetrizaci prostřednictvím multivariační logistické regrese s Firthovou korekcí. Výsledky: Celkem 10 931 pacientů podstoupilo transradiální a 3 208 transfemorální katetrizaci. Periprocedurální TIA/cévní mozková příhoda se vyskytla u 41 (0,29 %) pacientů. Věk byl jediným významným prediktorem TIA/mrtvice v multivariační analýze, přičemž každý další rok představoval poměr šancí (OR) = 1,09 (CI 1,05-1,13, p < 0,000). Volba místa nástupu neměla žádný vliv na riziko periprocedurální TIA/mrtvice (OR = 0,81; CI 0,38-1,72, p = 0,577). Závěr: Observační údaje z velkého prospektivního registru naznačují, že místo přistoupení nemá žádný vliv na riziko periprocedurální TIA/mrtvice po srdeční katetrizaci