Horynová, E*
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Článek představuje hlavní specifika logopedické intervence u pacientů s poruchami polykání v prostředí následné péče. Pacienti hospitalizovaní na lůžkách následné a dlouhodobé péče jsou často polymorbidní, geriatricky křehcí, mnozí s kognitivním deficitem. Těžiště logopedické terapie dysfagie u těchto pacientů tkví především v pasivních rehabilitačních technikách a režimových opatřeních, pro jejichž úspěšnou realizaci je klíčová mezioborová spolupráce s lékaři, fyzioterapeuty a především s ošetřovatelským personálem, mnohdy i s rodinnými příslušníky. Hlavním cílem příspěvku je na základě osobní profesní zkušenosti autorky podrobněji analyzovat již zmiňovaná specifika a představit hlavní osvědčené postupy logopedické intervence u pacientů s dysfagií v následné péči.
The article presents main specifics of speech and language therapy for patients with dysphagia at post-acute care. Patients hospitalized at post-acute and long-term care often suffer from polymorbidity, geriatric frailty, many have cognitive deficit. The core of speech and language therapy of dysphagia for these patients is mainly passive rehabilitation techniques and treatment measures. The interdisciplinary cooperation with physicians, physiotherapists and mainly nurses and attendants and often also family members is crucial for successful implementation of the measures. The main aim of the article is to analyse more in detail, on the grounds of personal professional experience of the author, the above mentioned specifics and introduce main time-tested methods of speech and language therapy for patients with dysphagia at post-acute care.
- MeSH
- křehký senior MeSH
- lidé MeSH
- následná péče MeSH
- poruchy polykání * diagnóza rehabilitace terapie MeSH
- řečová terapie * MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- senioři MeSH
- Publikační typ
- přehledy MeSH
- Klíčová slova
- ergodiagnostika,
- MeSH
- lidé MeSH
- nezávislé lékařské vyšetření MeSH
- posouzení stavu pacienta MeSH
- posuzování pracovní neschopnosti * MeSH
- posuzování zdravotní způsobilosti MeSH
- rehabilitace pracovní * MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
The human UDP-N-acetyl-α-d-galactosamine:polypeptide N-acetylgalactosaminyl-transferase 2 (GalNAc-T2) is one of the key enzymes that initiate synthesis of hinge-region O-linked glycans of human immunoglobulin A1 (IgA1). We designed secreted soluble form of human GalNAc-T2 as a fusion protein containing mouse immunoglobulin light chain kappa secretory signal and expressed it using baculovirus and mammalian expression vectors. The recombinant protein was secreted by insect cells Sf9 and human HEK 293T cells in the culture medium. The protein was purified from the media using affinity Ni-NTA chromatography followed by stabilization of purified protein in 50mM Tris-HCl buffer at pH 7.4. Although the purity of recombinant GalNAc-T2 was comparable in both expression systems, the yield was higher in Sf9 insect expression system (2.5mg of GalNAc-T2 protein per 1L culture medium). The purified soluble recombinant GalNAc-T2 had an estimated molecular mass of 65.8kDa and its amino-acid sequence was confirmed by mass-spectrometric analysis. The enzymatic activity of Sf9-produced recombinant GalNAc-T2 was determined by the quantification of enzyme-mediated attachment of GalNAc to synthetic IgA1 hinge-region peptide as the acceptor and UDP-GalNAc as the donor. In conclusion, murine immunoglobulin kappa secretory signal was used for production of secreted enzymatically active GalNAc-T2 in insect baculovirus expression system.
- MeSH
- aktivace enzymů MeSH
- Baculoviridae genetika metabolismus MeSH
- genetické vektory genetika metabolismus MeSH
- HEK293 buňky MeSH
- hmyz genetika metabolismus MeSH
- imunoglobulin A genetika metabolismus MeSH
- imunoglobuliny - kappa-řetězce chemie genetika MeSH
- klonování DNA MeSH
- kultivační média metabolismus MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši MeSH
- N-acetylgalaktosaminyltransferasy biosyntéza genetika izolace a purifikace MeSH
- plazmidy genetika metabolismus MeSH
- proteiny - lokalizační signály MeSH
- rekombinantní fúzní proteiny biosyntéza genetika izolace a purifikace MeSH
- rozpustnost MeSH
- sekvence aminokyselin MeSH
- stabilita proteinů MeSH
- tandemová hmotnostní spektrometrie MeSH
- transfekce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- srovnávací studie MeSH
Glycosylation abnormalities have been observed in autoimmune diseases and cancer. Here, we compare mechanisms of aberrant O-glycosylation, i.e., formation of Tn and sialyl-Tn structures, on MUC1 in breast cancer, and on IgA1 in an autoimmune disease, IgA nephropathy. The pathways of aberrant O-glycosylation, although different for MUC1 and IgA1, include dysregulation in glycosyltransferase expression, stability, and/or intracellular localization. Moreover, these aberrant glycoproteins are recognized by antibodies, although with different consequences. In breast cancer, elevated levels of antibodies recognizing aberrant MUC1 are associated with better outcome, whereas in IgA nephropathy, the antibodies recognizing aberrant IgA1 are part of the pathogenetic process.
- MeSH
- adenokarcinom imunologie MeSH
- glykosylace MeSH
- IgA nefropatie imunologie MeSH
- imunoglobulin A chemie imunologie MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- mucin 1 chemie imunologie MeSH
- nádory prsu imunologie MeSH
- polysacharidy chemie imunologie MeSH
- protilátky chemie imunologie MeSH
- prsy imunologie MeSH
- sacharidové sekvence MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Research Support, N.I.H., Extramural MeSH
Cysteine dioxygenase (CDO, EC 1.13.11.20) catalyses the oxygenation of cysteine to cysteine sulphinic acid leading to the production of sulphite, sulphate and taurine as the final metabolites of cysteine catabolism. Keratinolytic fungi secrete sulphite and sulphate to reduce disulphide bridges in host tissue keratin proteins as the first step of keratinolysis. In the present study, we describe the identification of cDNA, as well as expression and characterisation of recombinant CDO protein from Trichophyton mentagrophytes. The cDNA was amplified using primers designed on the basis of high conservancy CDO regions identified in other fungi. PCR product was cloned and sequenced. Recombinant CDO was expressed in Escherichia coli, and affinity purified and identified by matrix-assisted laser desorption/ionization - time-of-flight mass spectrometry (MALDI-TOF MS). Enzyme activity was assayed by monitoring the production of cysteine sulphinate using mass spectrometry. The Cdo cDNA encodes for a protein consisting of 219 amino acids. Recombinant CDO protein C-terminally fused with a His tag was purified by affinity chromatography. The CDO purified under native condition was proved to be enzymatically active. Protein identity was confirmed by MALDI-TOF MS. Comparison of cDNA sequence with those identified in other fungi revealed significant homology. Identification of T. mentagrophytes CDO provides indispensable tools for future studies of dermatophyte pathogenicity and development of new approaches for prevention and therapy.
- MeSH
- chromatografie afinitní MeSH
- cystein analogy a deriváty metabolismus MeSH
- cysteindioxygenasa genetika izolace a purifikace MeSH
- DNA fungální chemie genetika MeSH
- Escherichia coli genetika MeSH
- exprese genu MeSH
- klonování DNA MeSH
- komplementární DNA genetika MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- rekombinantní proteiny genetika izolace a purifikace metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie aminokyselin MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Trichophyton enzymologie genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- absorpční fotometrie MeSH
- lidé MeSH
- osteoporóza diagnóza MeSH
- rizikové faktory MeSH
- Check Tag
- lidé MeSH
Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-D-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite, N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other His.
- MeSH
- acetylglukosamin chemie metabolismus MeSH
- chromatografie afinitní MeSH
- endotoxiny biosyntéza izolace a purifikace MeSH
- Escherichia coli metabolismus MeSH
- glutaminfruktosa-6-fosfáttransaminasa (izomerizující) biosyntéza izolace a purifikace MeSH
- kultivační média chemie MeSH
- myši MeSH
- proteiny tepelného šoku HSP70 biosyntéza izolace a purifikace MeSH
- průmyslová mikrobiologie metody MeSH
- rekombinantní proteiny biosyntéza izolace a purifikace MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH