Hydrogen/deuterium exchange (HDX) is a well-established analytical technique that enables monitoring of protein dynamics and interactions by probing the isotope exchange of backbone amides. It has virtually no limitations in terms of protein size, flexibility, or reaction conditions and can thus be performed in solution at different pH values and temperatures under controlled redox conditions. Thanks to its coupling with mass spectrometry (MS), it is also straightforward to perform and has relatively high throughput, making it an excellent complement to the high-resolution methods of structural biology. Given the recent expansion of artificial intelligence-aided protein structure modeling, there is considerable demand for techniques allowing fast and unambiguous validation of in silico predictions; HDX-MS is well-placed to meet this demand. Here we present a protocol for HDX-MS and illustrate its use in characterizing the dynamics and structural changes of a dimeric heme-containing oxygen sensor protein as it responds to changes in its coordination and redox state. This allowed us to propose a mechanism by which the signal (oxygen binding to the heme iron in the sensing domain) is transduced to the protein's functional domain.
The nonradioactive method, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in the presence of Phos-tag (Phos-tag electrophoresis), is used to evaluate a kinase autophosphorylation and/or phosphotransfer reaction from a kinase/ATP to its protein substrate. This method outperforms radioisotope methods using [32P]ATP for detecting trace amounts of phosphorylated protein in fresh protein preparations. Phos-tag electrophoresis has been used to perform detailed analyses of the kinase activity of a heme-based oxygen sensor-specifically, a globin-coupled histidine kinase from the soil bacterium Anaeromyxobacter sp. Fw109-5 (AfGcHK).
The research focuses on the description of humour and its eff ect on drama-therapy intervention in people with addiction. In a short research, we used the so-called “sculpture” as one of the sources for obtaining data, where clients depicted selected thematic areas using their own bodies and subsequently verbally described their meaning. Another method used was a questionnaire survey with open questions, when clients answered thematic areas that arose from sculpture and last but not least it was fi eld observation of dramatherapists. This compares body-based data and subsequently ver-balises the attitudes and ideas of clients in conjunction with dramatherapist observa-tions. Dramatherapy as a discipline uses for its work means such as play, role play, improvisation and more. These means, and especially improvisation, off er a number of situations and stimuli where humour is revealed and applied. Th e topic of humour arises in almost every dramatherapy intervention and works in diff erent dimensions to the process. As it turns out, humour works on multiple levels, for example as an encouraging humour that helps build a therapeutic relationship, but also on opposite levels, serving as an escape from the topic the client is currently working on. It was the exploration of these levels of humour in dramatherapy intervention that was a central theme of the research study.
- MeSH
- Data Analysis MeSH
- Humans MeSH
- Substance-Related Disorders * psychology therapy MeSH
- Attitude MeSH
- Observation MeSH
- Surveys and Questionnaires MeSH
- Psychodrama methods MeSH
- Psychotherapists MeSH
- Laughter Therapy methods MeSH
- Hospitals, Psychiatric MeSH
- Wit and Humor as Topic * psychology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
Autoimunitní polyglandulární syndrom (APS) je heterogenní skupina onemocnění charakterizovaná autoimunitní reakcí proti více než jednomu endokrinnímu orgánu, přičemž mohou být postiženy i neendokrinní orgány. Prezentujeme případ desetiletého chlapce, který v současnosti trpí šesti autoimunitními nemocemi. Autoimunitní syndrom, typ 1, je velmi vzácné onemocnění, kdy je popsáno asi 500 případů na světě. Toto onemocnění se objevuje častěji u irských Židů, na Sardinii a u Finů. Dle našeho vyšetření nyní můžeme tvrdit, že se rovněž vyskytuje v České republice.
Autoimmune polyendocrine syndrome (APS) also called polyglandular autoimmune syndrome, are a heterogeneous group of rare diseases characterized by autoimmune activity against more than one endocrine organ, although non-endocrine organs can be affected. 10 years old boy suffers from 6 autoimmune diseases. Autoimmune polyglandular syndrome, type 1, is thought to be a rare condition, with about 500 cases reported worldwide. This condition occurs more frequently in certain populations, including Iranian Jews, Sardinians and Fins. According our examination we can confirm, that this type of syndrome exists also in Czech Republic.
- Keywords
- AIRE protein,
- MeSH
- Addison Disease diagnosis MeSH
- Polyendocrinopathies, Autoimmune * diagnosis genetics MeSH
- Thyroiditis, Autoimmune diagnosis MeSH
- Cytogenetic Analysis MeSH
- Diabetes Mellitus, Type 1 diagnosis MeSH
- Diagnosis, Differential * MeSH
- Child MeSH
- Humans MeSH
- Mutation MeSH
- Transcription Factors genetics MeSH
- Vitiligo MeSH
- Rare Diseases MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
Edukace
1. vyd. Nestr. : il. ; 21 cm
Povídání o diabetu a o dospívání pro patnáctileté i pro ty o trochu starší; Povídání o diabetu a o dospívání pro patnáctileté i pro ty trochu starší.
- MeSH
- Diabetes Mellitus, Type 1 MeSH
- Diabetes Mellitus MeSH
- Adolescent MeSH
- Psychology, Adolescent MeSH
- Check Tag
- Adolescent MeSH
- Publication type
- Poetry MeSH
- Popular Work MeSH
- Conspectus
- Patologie. Klinická medicína
- NML Fields
- diabetologie
- pediatrie
- NML Publication type
- literatura pro děti a mládež
Tranzitorní refrakční změny jsou dobře rozpoznatelné znaky diabetes mellitus a nejen oftalmologové, ale především pediatři by vždy měli pátrat po diabetu v případě rychlých refrakčních změn.
Transient refractive changes are well recognised features of diabetes and not only ophtalmologists but especially paediatricians should always check for diabetes mellitus in any case of rapidly changing refraction.
V pediatrické praxi se setkáváme s narůstající incidencí diabetes mellitus 1. typu. Značná část takto nově diagnostikovaných případů přichází již ve fázi pozdní diabetické ketoacidózy. Autoimunitní proces, který vyústí v klinicky manifestní diabetes mellitus však probíhá měsíce až roky před manifestací onemocnění. Jaké jsou možnosti naší včasné diagnostiky? Je možné diabetes zachytit v preklinické fázi? Kdy je možno uvažovat o diagnóze MODY? Při pandemii obezity musíme i u dětí stále častěji zvažovat možnost diabetes mellitus 2. typu.
In paediatrics we face growing incidence of diabetes mellitus type 1. Considerable part of such diagnosed cases come too late, in the phase of diabetic ketoacidosis. Self-immunity process, which leads into clinically manifesting diabetes mellitus, proceeds for months even years. What are the chances of our early diagnosis? Is it possible to notice the diagnosis in pre-clinical phase? When should we consider diagnosis called MODY? In connection with obesity we must think about diabetes mellitus type 2 in childhood as well.
- MeSH
- Diabetes Mellitus, Type 1 diagnosis genetics pathology MeSH
- Diabetes Mellitus, Type 2 diagnosis genetics pathology MeSH
- Diagnostic Techniques, Endocrine MeSH
- Genetic Vectors MeSH
- Hyperglycemia diagnosis pathology MeSH
- Humans MeSH
- Adolescent MeSH
- Obesity pathology MeSH
- Check Tag
- Humans MeSH
- Adolescent MeSH
- Male MeSH
- Publication type
- Case Reports MeSH
