Characterization of cellular stress systems using biological mass spectrometry

In recent years mass spectrometry has become an invaluable tool to address an array of biological questions. The versatility of this experimental approach does not only allow assignment of protein identity and identification of sequence specific modifications, but with the help of particular derivatization techniques facilitates the determination of peptide quantity. Each of these approaches were applied to the following biological projects: The 21 kDa heat stable protein purified from the encysted embryo of Artemia franciscana was characterized by time-of-flight mass spectrometry. De novo sequencing of peptides identified this protein as a group 1 Late embryogenesis abundant (LEA) protein. The amino acid sequence assignment to these peptides allowed amplification of the entire gene sequence from an embryonic cDNA library. This was deposited into the NCBI database (EF656614). The expression of group 1 LEA protein is consistent with and supports a role in desiccation tolerance. In addition, this is a first report describing identification of a group 1 LEA protein in an animal species. A MS-based quantitative analysis was performed in order to analyze relative changes in the dynamic thiol and disulfide states of the redox sensitive protein disulfide isomerase, PDI. PDI cysteine residues were derivatized with an isotope-coded affinity tag (ICAT), thus allowing a direct comparison between the reduced and auto-oxidized forms. Quantitation was based on relative ratios between light and heavy isotopically labeled cysteine containing peptides. The application of the ICAT-labeling approach to PDI related studies, allowed direct assignment of individual cysteine residues and their oxidation status, compared to the previously employed techniques, that only provided information regarding the average number of modified cysteine residues within PDI, not their exact identity. A combination of a phosphopeptide enrichment step and a MS-based approach was utilized to identify three phosphorylation sites on hYVH1, an atypical dual specificity phosphatase that functions as a cell survival factor. With the help of phosphomimetic and non-phosphorylable mutants, we were able to decipher their effect on localization and progression through the cell cycle. Collectively, these studies manifest the power of MS-generated data to influence and guide many different fields ranging from molecular embryology to biochemistry

Level of auditory analysis, synthesis and active vocabulary and their intergender context : [Uroven sluchovej analyzy, syntezy a aktivna slovna zasoba a ich mezipohlavny kontext

The subject of our research were partial cognitive functions and phonematic awareness of preschool students. Following the findings, we focused on the identification of selected determinants of auditory analysis and synthesis, namely gender differences in the level of auditory analysis and active vocabulary of six-year-old children. Our aim was also to find out whether there is a difference in the level of auditory analysis and auditory synthesis in six-year-old

Evaluation of a Bacillus stearothermophilus tube test as a screening tool for anticoccidial residues in poultry

A Bacillus stearothermophilus var. calidolactis C953 tube test was evaluated for its ability in detecting the residue of selected anticoccidial drugs in poultry, specically sulfamethazine, furazolidone, and amprolium. Various concentrations of each drug were injected into chicken liver and kidney tissues and these tissues were tested to determine the drug detection limits for each drug. The detection limit was defined as the drug concentration at which 95% of the test results were interpreted as positive. The limits of detection in liver tissue were 0.35 µg/ml for furazolidone, 0.70 µg/ml for sulfamethazine and 7.80 µg/ml for amprolium. In kidney tissues, they were 0.30 µg/ml for furazolidone, 0.54 µg/ml for sulfamethazine, and 7.6 µg/ml for amprolium. It was concluded that this tube test could be used to screen for the residue of these three drugs in poultry

Efficacy of Chondroprotective Food Supplements Based on Collagen Hydrolysate and Compounds Isolated from Marine Organisms †

Osteoarthritis belongs to the most common joint diseases in humans and animals and shows increased incidence in older patients. The bioactivities of collagen hydrolysates, sulfated glucosamine and a special fatty acid enriched dog-food were tested in a dog patient study of 52 dogs as potential therapeutic treatment options in early osteoarthritis. Biophysical, biochemical, cell biological and molecular modeling methods support that these well-defined substances may act as effective nutraceuticals. Importantly, the applied collagen hydrolysates as well as sulfated glucosamine residues from marine organisms were strongly supported by both an animal model and molecular modeling of intermolecular interactions. Molecular modeling of predicted interaction dynamics was evaluated for the receptor proteins MMP-3 and ADAMTS-5. These proteins play a prominent role in the maintenance of cartilage health as well as innate and adapted immunity. Nutraceutical data were generated in a veterinary clinical study focusing on mobility and agility. Specifically, key clinical parameter (MMP-3 and TIMP-1) were obtained from blood probes of German shepherd dogs with early osteoarthritis symptoms fed with collagen hydrolysates. Collagen hydrolysate, a chondroprotective food supplement was examined by high resolution NMR experiments. Molecular modeling simulations were used to further characterize the interaction potency of collagen fragments and glucosamines with protein receptor structures. Potential beneficial effects of collagen hydrolysates, sulfated glycans (i.e., sulfated glucosamine from crabs and mussels) and lipids, especially, eicosapentaenoic acid (extracted from fish oil) on biochemical and physiological processes are discussed here in the context of human and veterinary medicine

Regulation of neovascularization by S-glutathionylation via the Wnt5a/sFlt-1 pathway

S-glutathionylation occurs when reactive oxygen or nitrogen species react with protein-cysteine thiols. Glutaredoxin-1 (Glrx) is a cytosolic enzyme which enzymatically catalyses the reduction in S-glutathionylation, conferring reversible signalling function to proteins with redox-sensitive thiols. Glrx can regulate vascular hypertrophy and inflammation by regulating the activity of nuclear factor κB (NF-κB) and actin polymerization. Vascular endothelial growth factor (VEGF)-induced endothelial cell (EC) migration is inhibited by Glrx overexpression. In mice overexpressing Glrx, blood flow recovery, exercise function and capillary density were significantly attenuated after hindlimb ischaemia (HLI). Wnt5a and soluble Fms-like tyrosine kinase-1 (sFlt-1) were enhanced in the ischaemic-limb muscle and plasma respectively from Glrx transgenic (TG) mice. A Wnt5a/sFlt-1 pathway had been described in myeloid cells controlling retinal blood vessel development. Interestingly, a Wnt5a/sFlt-1 pathway was found also to play a role in EC to inhibit network formation. S-glutathionylation of NF-κB components inhibits its activation. Up-regulated Glrx stimulated the Wnt5a/sFlt-1 pathway through enhancing NF-κB signalling. These studies show a novel role for Glrx in post-ischaemic neovascularization, which could define a potential target for therapy of impaired angiogenesis in pathological conditions including diabetes

A Drosophila-centric view of protein tyrosine phosphatases

AbstractMost of our knowledge on protein tyrosine phosphatases (PTPs) is derived from human pathologies and mouse knockout models. These models largely correlate well with human disease phenotypes, but can be ambiguous due to compensatory mechanisms introduced by paralogous genes. Here we present the analysis of the PTP complement of the fruit fly and the complementary view that PTP studies in Drosophila will accelerate our understanding of PTPs in physiological and pathological conditions. With only 44 PTP genes, Drosophila represents a streamlined version of the human complement. Our integrated analysis places the Drosophila PTPs into evolutionary and functional contexts, thereby providing a platform for the exploitation of the fly for PTP research and the transfer of knowledge onto other model systems

Innowacyjne i informacyjne perspektywy zarządzania przedsiębiorstwem

The purpose of this study is to point out the extent to which enterprises want to exploit the possibilities of information technologies (IT), system integration of information systems (IS) and business process automation with the aim to increase the intelligent work with information and improve business management support in enterprises. The paper evaluates the development and analyzes the views of 189 managers of Slovak SMEs and 26 managers of IT companies on the causes that prevent wider use of innovation opportunities and IS potential in enterprises in the process of increasing the competitiveness of their enterprises in the market. In this case we focused on the analysis and identification of several advantages and disadvantages related to the implementation of innovation and process automation and their impact on business management. We compare these results with the results of similar researches in the EU. Based on the best practice results, we suggest practices and recommendations that should eliminate the barriers to successful implementation of innovation potential in businesses and create conditions to support managerial decision making processes to be more effective.Celem niniejszego artykułu jest wskazanie, w jakim stopniu przedsiębiorstwa chcą wykorzystać możliwości technologii informacyjnych (IT), systemowej integracji systemów informatycznych (IS) i automatyzacji procesów biznesowych w celu zwiększenia inteligentnej pracy z informacjami oraz poprawy wsparcia zarządzania w przedsiębiorstwach. Artykuł ocenia rozwój i analizuje opinie 189 menedżerów słowackich MŚP i 26 menedżerów firm IT na temat przyczyn, które uniemożliwiają szersze wykorzystanie możliwości innowacyjnych i potencjału systemów informatycznych w przedsiębiorstwach w procesie zwiększania konkurencyjności na rynku. W tym przypadku skupiono się na analizie i identyfikacji szeregu zalet i wad związanych z wdrażaniem innowacji i automatyzacji procesów oraz ich wpływem na zarządzanie przedsiębiorstwem. Porównano osiągnięte wyniki z rezultatami podobnych badań w UE. W oparciu o wyniki najlepszych praktyk zaproponowano rozwiązania i zalecenia, które powinny wyeliminować bariery utrudniające skuteczne wdrażanie potencjału innowacyjnego w przedsiębiorstwach i stworzyć warunki do wspierania procesów decyzyjnych menedżerów, tak aby były bardziej skuteczne

A rare case of hard palate radicular cyst

Inflammatory odontogenic cysts comprise the entity of radicular cysts which may affect bone jaws to a different extend. Most of these lesions encompass the highest structure – the dental apex and are well defined and delineated from adjacent benign structures. Concerning different clinical characteristics of the remaining odontogenic cysts differential diagnosis includes: residual cyst, follicular cyst, keratocyst, non-odontogenic fissural cysts as well as some malignant diseases including ameloblastoma, cementoma, Pindborg tumor, etc. This article presents a sole case of a massive odontogenic radicular cyst, affecting the right half of the hard palate at the area of lacking molars, crossing midline and containing dental apex. The case was diagnosed as infected radicular cyst on the basis of clinical, histopathological and radiological findings. The clinical characteristics of this cyst represent an interesting and extraordinary finding due to the presence of a foreign body in it, situated far from the upper left side teeth, extracted several years before its appearance. The lesion was surgically enucleated including the apex when all adjacent teeth and vital oral structures were kept intact. No postoperative complications were endured by the patient and normal healing process wasachieved