V poslední době se intenzivně diskutuje potřeba změn prostředí v psychiatrii - nejen prostředí, ve kterém pacient přechodně žije (zde je myšlen zejména ubytovací standard v psychiatrických nemocnicích), ale i prostředí, ve kterém terapie probíhá (myšleny nejen parametry prostoru, ale celková atmosféra a podoba vztahů a také postavení člověka v tomto prostředí). V našem článku se budeme věnovat specificky léčbě závislostí, byť základní teze a závěry lze přenést na celou psychiatrii.
In recent years, one of the important topics of neurobiological research is environmental enrichment. A positive and inspiring environment with sensory and motor inspirations and increasing well-being of the organism can significantly repair mental disorders and improve the condition of the brain. In addition to the obviously effective therapeutic moments, our traditional addiction treatment programs have contained "stenizing" elements and elements that are completely opposite to the concept of environmental enrichment. Eliminating these distressing elements and consistently working with environmental enrichment could make our inpatient treatment programs more effective.
Rapid changes of protein phosphorylation play a crucial role in the regulation of many cellular processes. Being post-translationally modified, phosphoproteins are often present in quite low abundance and tend to co-exist with their unphosphorylated isoform within the cell. To make their identification more practicable, the use of enrichment protocols is often required. The enrichment strategies can be performed either at the level of phosphoproteins or at the level of phosphopeptides. Both approaches have their advantages and disadvantages. Most enriching strategies are based on chemical modifications, affinity chromatography to capture peptides and proteins containing negatively charged phosphate groups onto a positively charged matrix, or immunoprecipitation by phospho-specific antibodies.In this article, the most up-to-date enrichment techniques are discussed, taking into account their optimization, and highlighting their advantages and disadvantages. Moreover, these methods are compared to each other, revealing their complementary nature in providing comprehensive coverage of the phosphoproteome.
- MeSH
- Staining and Labeling MeSH
- Chromatography, Affinity MeSH
- Phosphoproteins chemistry isolation & purification metabolism MeSH
- Phosphorylation MeSH
- Mass Spectrometry MeSH
- Immunoprecipitation MeSH
- Peptide Fragments chemistry isolation & purification metabolism MeSH
- Protein Processing, Post-Translational MeSH
- Proteome chemistry isolation & purification metabolism MeSH
- Proteomics MeSH
- Plant Proteins chemistry isolation & purification metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
Článek představuje teoretické pozadí metody instrumentálního obohacování Reuvena Feuersteina a její využití v praxi. Pojednává o životě autora, jeho teorii strukturální kognitivní modifikovatelnosti a zkušenosti zprostředkovaného učení. Dále představuje využití metody ve třech oblastech – školní, sociální a oblasti zdravotnictví. Ve školním prostředí se článek zaměřuje na využití u žáků s ADHD, specifickými poruchami učení, sociálním znevýhodněním a na rozvoj učitelů. V sociální oblasti představuje možnosti metody k intervenci u seniorů a dospělých, dále také u osob ve výkonu trestu odnětí svobody. V oblasti zdravotnictví představuje její aplikaci u osob s traumatickým poškozením mozku a Parkinsonovou nemocí.
This article presents the theoretical background of Reuven Feuerstein ́s Instrumental Enrichment and its application. It discusses Feuerstein ́s life, his theory of structural cognitive modifiability, and mediated learning experience. It shows the application of the program in three areas – education, social area, and healthcare. In the area of education, it discusses the application in students with ADHD, specific learning disabilities, social disadvantage, and a teacher ́s development. In the social area, it presents the application of the program in seniors, adults, and at the prison. In the area of healthcare, application in people with traumatic brain injury and Parkinson ́s disease is presented.
- Keywords
- metoda instrumentálního obohacování, Feursteinova metoda,
- MeSH
- Attention Deficit Disorder with Hyperactivity MeSH
- Cognition MeSH
- Humans MeSH
- Neuronal Plasticity MeSH
- Brain Injuries rehabilitation MeSH
- Education, Special * history methods MeSH
- Learning MeSH
- Education * history methods MeSH
- Child Development MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Cíl studie: Koncentrace jaderných erytroblastů cirkulujících v periferní krvi gravidních žen proúčely neinvazivní prenatální diagnostiky.Typ studie: Pilotní studie.Název a sídlo pracoviště: II. dětská klinika 2. LF UK, Fakultní nemocnice Motol, Praha, Českárepublika.Metodika: Z 13 - 28 ml periferní krve 78 gravidních žen v rozmezí 13. - 37. týdnu gravidity bylyizolovány mononukleární buňky. Mateřské leukocyty byly odstraněny z frakce mononukleárníchbuněk pomocí anti-CD14 a anti-CD45 monoklonálních protilátek značených superparamagnetic-kými částicemi metodou magnetické separace buněk (MACS) na zařízení VarioMACS. Jadernéerytroblasty byly následně koncentrovány z CD14 - /CD45 - frakce pozitivní selekcí pomocí anti-CD71 monoklonální protilátky na zařízení MiniMACS. Jednotlivé frakce byly analyzovány tříba-revnou analýzou na průtokovém cytometru (FACS).Výsledky: U 68 těhotných žen z 78 testovaných (87 %) byly nalezeny po dvojité magnetické separa-ci buněk (doubleMACS) zahrnující depleci leukocytů a pozitivní selekci jaderné erytroblastyv rozmezí 2x10 5 - 1,02 x 10 6 . Faktor celkového zkoncentrování se pohyboval v rozmezí 4,2 - 526,0(Ć 138,4). Z našeho souboru vyšetřených je patrné, že počet CD71 + jaderných erytroblastů koncen-trovaných metodou doubleMACS je značně individuální. Jaderné erytroblasty cirkulující v krev-ním oběhu gravidních žen bylo možno zkoncentrovat již v 13. týdnu gravidity.Závěr: Naším cílem je vypracovat a standardizovat metodu koncentrace jaderných erytroblastůcirkulujících v periferii gravidních žen a ověřit možnost použití tohoto alternativního zdroje proúčely neinvazivní prenatální diagnostiky.
Objective: Enrichment of nucleated red blood cells (NRBCs) from maternal blood for non-invasiveprenatal diagnosis.Design: Pilot study.Setting: 2 nd Clinic of Paediatrics, University Hospital Motol, Prague, Czech Republic.Methods: Mononuclear cells were isolated from 13 - 28 ml of peripheral maternal blood between 13and 37 weeks of gestation. Leukocytes from maternal peripheral blood were depleted from mono-nuclear cells by treatment with anti-CD14 and anti-CD45 microbeads and high-gradient magneticcell separation (MACS) on VarioMACS. NRBCs were sorted from CD14 - /CD45 - fraction by positiveselection using anti-CD71 microbeads on MiniMACS. All sorting steps were analysed by three-co-lour cytometric analysis with FACScan flow cytometer.Results: In 68 out of 78 pregnant woman (87 %) NRBCs were found in range 2x10 5 - 1,02 x 10 6.NRBC were enriched with an average enrichment rate of 138-fold ranging from 4 - 526 fold. In ourcohort of pregnant woman the number of isolated NRBCs was individual. We identified NRBCsfrom the 13 th week of gestation.Conclusion: The aim of the study is to establish and standardise the method of enrichment ofNRBCs from maternal blood samples and verify the applicability of this alternative source fornon-invasive prenatal diagnosis.
- MeSH
- Biological Transport MeSH
- Adult MeSH
- Erythroblasts blood MeSH
- Humans MeSH
- Maternal-Fetal Exchange MeSH
- Antibodies, Monoclonal diagnostic use MeSH
- Prenatal Diagnosis MeSH
- Flow Cytometry methods instrumentation MeSH
- Pregnancy MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Review MeSH
Phosphorylation is among the most important post-translational modifications of proteins and has numerous regulatory functions across all domains of life. However, phosphorylation is often substoichiometric, requiring selective and sensitive methods to enrich phosphorylated peptides from complex cellular digests. Various methods have been devised for this purpose and we have recently described a Fe-IMAC HPLC column chromatography setup which is capable of comprehensive, reproducible, and selective enrichment of phosphopeptides out of complex peptide mixtures. In contrast to other formats such as StageTips or batch incubations using TiO2or Ti-IMAC beads, Fe-IMAC HPLC columns do not suffer from issues regarding incomplete phosphopeptide binding or elution and enrichment efficiency scales linearly with the amount of starting material. Here, we provide a step-by-step protocol for the entire phosphopeptide enrichment procedure including sample preparation (lysis, digestion, desalting), Fe-IMAC column chromatography (column setup, operation, charging), measurement by LC-MS/MS (nHPLC gradient, MS parameters) and data analysis (MaxQuant). To increase throughput, we have optimized several key steps such as the gradient time of the Fe-IMAC separation (15 min per enrichment), the number of consecutive enrichments possible between two chargings (>20) and the column recharging itself (<1 h). We show that the application of this protocol enables the selective (>90 %) identification of more than 10,000 unique phosphopeptides from 1 mg of HeLa digest within 2 h of measurement time (Q Exactive Plus).
- MeSH
- Cell Line MeSH
- Chromatography, Liquid MeSH
- Phosphopeptides MeSH
- Phosphoproteins * MeSH
- Imidazoles chemistry MeSH
- Humans MeSH
- Proteome * MeSH
- Proteomics methods MeSH
- Workflow MeSH
- Software MeSH
- Statistics as Topic MeSH
- Tandem Mass Spectrometry MeSH
- Iron chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Protein phosphorylation was repeatedly shown to be the most dynamic post-translational modification mediated by a huge orchestra of protein kinases and phosphatases. Upon landing on a stigma, pollen grain dehydration and activation are accompanied by changes in protein phosphorylation together with the translation activation of stored mRNAs. To enable studies of the total phosphoproteome, it is usually necessary to apply various enrichment techniques. In this chapter, one of these protocols that worked previously well on tobacco mature pollen is presented in more detail. The method comprises of three basic steps: (1) picking flowers from the flowering tobacco plants (Nicotiana tabacum cv. Samsun), and collection of the shed pollen grains; (2) extraction of total proteins by TCA/acetone; (3) phosphoprotein enrichment by MOAC with aluminum hydroxide matrix. Taken together this protocol describes how to isolate phosphoproteins out of tobacco mature pollen.
- MeSH
- Chromatography, Affinity MeSH
- Phosphoproteins chemistry isolation & purification metabolism MeSH
- Aluminum Hydroxide chemistry MeSH
- Protein Processing, Post-Translational MeSH
- Pollen metabolism MeSH
- Plant Proteins chemistry isolation & purification metabolism MeSH
- Nicotiana metabolism MeSH
- Publication type
- Journal Article MeSH
The potential oil-producing yeast Rhodotorula gracilis was found to produce higher yields of biomass (13.7 g/L) and lipids (20.3%) in a nitrogen-limited and economically cheaper medium (molasses without yeast extract) in a submerged fermentation system. But, when the yeast was grown on four different wheat bran media by solid-state fermentation technique, different media combinations affected the percent increase in biomass, protein, oil production, fatty acid profile and degree of saturation and unsaturation. The initial lipid content in the control medium was 3.5% while in a medium with wheat bran, molasses, and minerals it was 69.8%. The yeast did not produce alpha-amylase, amyloglucosidase and cellulolytic enzymes for the breakdown of wheat bran. The yeast produced red carotenoids, a precursor of vitamin B12 and some oligounsaturated fatty acids in the fermented product.
- MeSH
- Biotechnology MeSH
- Fermentation MeSH
- Food, Fortified MeSH
- Culture Media MeSH
- Lipids biosynthesis MeSH
- Fatty Acids metabolism MeSH
- Carbohydrate Metabolism MeSH
- Mycology methods MeSH
- Oils metabolism MeSH
- Food Technology MeSH
- Dietary Fiber metabolism MeSH
- Rhodotorula growth & development metabolism MeSH
- Publication type
- Journal Article MeSH
