Characterisation of Extracellular Matrix Cross-Linking in Systemic Sclerosis

Systemic sclerosis (SSc) is a complex connective tissue disease associated with a high mortality. As development of skin fibrosis in SSc patients leads to major organ-based complications, finding a treatment that resolves this condition can help prevent disease progression. A major obstacle in the discovery of new effective anti-fibrotic therapies is that the exact mechanisms that lead to fibrotic progression and irreversibility have not been fully elucidated. The fibrotic phenotype is associated with increased accumulation of extracellular matrix (ECM) in an organ that severely impedes its function. Excessive cross-linking of the ECM is a major factor causing its resistance to remodelling by rendering sites of proteolysis less accessible to matrix metalloproteinases. The discovery of new cross-linking sites during disease progression may therefore improve our molecular understanding of the resistance of the ECM to proteolytic breakdown, act as a clinical marker for resolvable vs non-resolvable fibrosis, and contribute to the development of new therapeutic approaches. Transglutaminases are a group of cross-linking enzymes that play a pivotal role in fibrotic progression. A mass spectrometry-based method for the identification of transglutaminase cross-linking sites from complex biological samples has been developed in this project. A model transglutaminase cross-linked peptide was synthesized and its fragmentation behaviour was analysed. After having found the optimal fragmentation conditions the analysis of cross-linking sites was automated using MassMatrix software. The resulting score values were utilised to identify true-positive matches. This technique was successfully applied to an analysis of the transglutaminase cross-linking site of purified fibrinogen dimers. In the final stages of the method development comparative studies of the ECM cross-linking pattern between control and pro-fibrotic dermal fibroblast cell culture and mouse models were performed. The limited number of new cross-linking sites found in the fibrotic models suggest that more significant changes occur at later fibrotic stages

INVESTIGATION ON SOME MORPHOLOGICAL AND BIOLOGICAL CHARACTERISTICS OF EINKORN WHEAT (T. MONOCOCCUM L.) DEPENDING ON NITROGEN FERTILIZATION

The aim of this study is to investigate some quantitative and qualitative indicators of einkorn wheat (T. monococcum L.). A three-year field experiment has been carried out at the experimental field of Department of Crop Science in Agricultural University - Plovdiv. To compare the performance, Sadovo1 common wheat (T. aestivum L.) is used as a standard. Both wheat species have been grown on two nitrogen fertilization levels – 80 and 160 kg.ha-1 nitrogen. Phenological development of the plants was recorded at the onset of the main phenophase. Inter-phase period has been calculated (number of days). Grain yield (t ha-1) is accounted indirectly by ¼ m2 plot. The main structural elements of plants have been established. It has been found that phenological development stage of tillering occurs at the same time for both wheat species. Following the start of spring vegetation, common wheat enters a phase earlier than the einkorn. Common wheat is a high-yielding einkorn, that puts both proven wheat varieties in different groups. Einkorn has high tiller appearance but it has a low productive tillering than common wheat. Einkorn form lower grain in the spike and lighter grain per spike. Nitrogen fertilization significantly increased harvested grain in common wheat. In einkorn it has no significant impact on yield

Міжнародна економічна діяльність країни в умовах глобального фінансового конкурентного розвитку

У статті проведено комплексне наукове обґрунтування теоретичних і практичних засад становлення міжнародної економічної діяльності країни в умовах глобального фінансового конкурентного розвитку. (In the article it is conducted complex scientific ground of theoretical and practical principles of becoming of international economic activity of country in the conditions of global financial competition development.

Mass Spectrometric Identification of a Novel Factor XIIIa Cross-Linking Site in Fibrinogen

Transglutaminases are a class of enzymes that catalyze the formation of a protein:protein cross-link between a lysine and a glutamine residue. These cross-links play important roles in diverse biological processes. Analysis of cross-linking sites in target proteins is required to elucidate their molecular action on target protein function and the molecular specificity of different transglutaminase isozymes. Mass-spectrometry using settings designed for linear peptide analysis and software designed for the analysis of disulfide bridges and chemical cross-links have previously been employed to identify transglutaminase cross-linking sites in proteins. As no control peptide with which to assess and improve the mass spectrometric analysis of TG cross-linked proteins was available, we developed a method for the enzymatic synthesis of a well-defined transglutaminase cross-linked peptide pair that mimics a predicted tryptic digestion product of collagen I. We then used this model peptide to determine optimal score thresholds for correct peptide identification from y- and b-ion series of fragments produced by collision-induced dissociation. We employed these settings in an analysis of fibrinogen cross-linked by the transglutaminase Factor XIIIa. This approach resulted in identification of a novel cross-linked peptide in the gamma subunit. We discuss the difference in behavior of ions derived from different cross-linked peptide sequences and the consequent demand for a more tailored mass spectrometry approach for cross-linked peptide identification compared to that routinely used for linear peptide analysis

TGFβ-1 Induced Cross-Linking of the Extracellular Matrix of Primary Human Dermal Fibroblasts

Excessive cross-linking is a major factor in the resistance to the remodelling of the extracellular matrix (ECM) during fibrotic progression. The role of TGFβ signalling in impairing ECM remodelling has been demonstrated in various fibrotic models. We hypothesised that increased ECM cross-linking by TGFβ contributes to skin fibrosis in Systemic Sclerosis (SSc). Proteomics was used to identify cross-linking enzymes in the ECM of primary human dermal fibroblasts, and to compare their levels following treatment with TGFβ-1. A significant upregulation and enrichment of lysyl-oxidase-like 1, 2 and 4 and transglutaminase 2 were found. Western blotting confirmed the upregulation of lysyl hydroxylase 2 in the ECM. Increased transglutaminase activity in TGFβ-1 treated ECM was revealed from a cell-based assay. We employed a mass spectrometry-based method to identify alterations in the ECM cross-linking pattern caused by TGFβ-1. Cross-linking sites were identified in collagens I and V, fibrinogen and fibronectin. One cross-linking site in fibrinogen alpha was found only in TGFβ-treated samples. In conclusion, we have mapped novel cross-links between ECM proteins and demonstrated that activation of TGFβ signalling in cultured dermal fibroblasts upregulates multiple cross-linking enzymes in the ECM

Higher education: problems and opportunities

В статье обосновано, что современная молодежь стремится к получению высшего образования, так как профессиональная карьера невозможна без качественного высшего образования, которое может быть получено, как в российских, так и зарубежных университетах.In the article it is justified that the contemporary youth is striving for higher education, because it is impossible to make a professional career without a high-quality education that can be obtained both at Russian and foreign universities

Pohyby migrantov z metropoly východu v globalizujúcom sa svete

This paper is focused on current trends in migration processes. The content is based on results of qualitative research, using the grounded theory method. This approach followed the movements of young migrants from Eastern Slovak city Košice, in different phases of development. Findings of the research were tied with relevant theories describing current processes, which are the results of intensive changes in society and create challenging environment for migration. Movements of migrants are directly connected to life perspectives and strategies in creating an optimal life conditions. The goal of the paper is to reveal behavioral similarities of migrant, in individualized and diversificated society, and to bring more comprehensive view of migration issue.Táto diplomová práca je zameraná na problematiku súčastných trendnov v migrančých procesoch. Jej obsah sa odvíja od výsledkov kvalitatívneho výskumu, prevedeného prostredníctvom metody grounded theory. Tímto prístupom boli sledované pohyby mladých migrantov z východnej metropoly Slovenska, Košíc, v rôznych štádiách posunu. Prežité udalosti týchto aktérov sú previazané z relevantnými teóriami pokúšajúce sa popisovať súčasné procesy, ktoré ako výsledok intentízvnejších premien prenikajúcich spoločnosťou a vytvárajú podnetné prostredie pre pohyb obyvateľstva za hranice štátov. Smer migrácie skúmaných aktérov ma priamu náväznosť na stavbu životnej perspektívny a vyvíjaniu stratégií v hladaní optimálnych podmienok. Cieľom práce je nachádzať podobnosti správania sa jedincov inak individualizovanej či diverzifikovanej spoločnosti a priniesť celistvejší náhľad na problematiku migrácieDepartment of Historical SociologyPracoviště Historické sociologieFaculty of HumanitiesFakulta humanitních studi

Uncertainties of calculated coincidence-summing correction factors in gamma-ray spectrometry

Uncertainty propagation to the $\gamma$-$\gamma$ coincidence-summing correction factor from the covariances of the nuclear data and detection efficiencies have been formulated. The method was applied in the uncertainty analysis of the coincidence-summing correction factors in the $\gamma$-ray spectrometry of the $^{134}$Cs point source using a p-type coaxial HPGe detector.Comment: 4 pages, 2 figures, to be published in the proceedings of the "2019 International Conference on Nuclear Data for Science and Technology" (ND2019

UV-triggered syndrome of Gougerot-Carteaud - mechanical damage or immunological reaction

Papillomatosis papulosa confluens reticulata et pigmentata is a rare dermatosis that presents clinically with pigmented, papillomatous, hyperkeratotic plaques with central confluence and reticulated peripheral pattern. The etiology remains unclear but current data points to a keratinization defect as the main pathological mechanism. There is one case report in the literature of a UV-induced Papillomatosis papulosa confluens reticulata et pigmentata. We present a 16-year old patient with clinical and histological diagnosis of confluent and reticulated papillomatosis. The characteristic lesions involved sun-exposed areas and developed after intense sun exposure. As an etiologic factor, ultraviolet light may lead to mechanical epidermal damage and further keratinization modification. On the other hand, in the setting of genetically predetermined resistance to UV-immunosuppression mechanisms, the newly formed epidermal antigens could potentially trigger an immune reaction resulting in skin eruption.Scripta Scientifica Medica 2013; 45(4): 88-91

Acute generalized exanthematous pustulosis (AGEP): a literature review

Acute generalized exanthematous pustulosis (AGEP) is a rare drug-induced pustular cutaneous reaction. The clinical course is characteristic with a sudden onset of multiple sterile pustules on an erythematous base with fever and neutrophilia, followed by a spontaneous resolution within two weeks. Drug-specific T cells play the main role in the pathogenesis, but the exact cytokine cascade and genetic background are yet to be elucidated. Timely and exact recognition is important in order to prevent confusion with infections and psoriasis and hence institution of unnecessary and wrong treatments. The diagnosis may be confirmed by typical history, identification of a culprit drug, histopathology and patch testing. The purpose of this review is to present the current knowledge on AGEP and its association with various drugs in the context of a drug allergic reaction.Scripta Scientifica Medica 2013; 45(4): 7-12