Using of NTC method to the development of partial functions of preschool children

V diplomové práci se věnuji rozvoji dovedností dětí předškolního věku v předškolním zařízení. Práce je rozdělena na teoretickou a praktickou část. Kapitoly teoretické části se zabývají charakteristikou vývoje dítěte v předškolním věku, rozvojem dílčích funkcí a osobnostním vývojem a výchovou dítěte předškolního věku. Zabývá se vzděláváním dětí v mateřských školách a otázkou školní zralosti. Praktická část diplomové práce je zaměřena na zavedení stimulačního programu NTC do výchovně vzdělávací činnosti soukromého vzdělávacího centra Salomon Preschool pro předškolní děti. V této části je zhodnocena metoda NTC na základě komparace výzkumné a kontrolní skupiny tvořené dětmi ze státní mateřské školy a dále jsou zde navrženy vhodné aktivity pro předškolní děti. Navržené aktivity jsou zaměřené na podporu konkrétních klíčových kompetencí a dovedností. KLÍČOVÁ SLOVA Dílčí funkce, předškolní věk, vývoj dítěte v předškolním věku, možnosti rozvoje dílčích funkcí, školní zralost, stimulace, vzdělávání dětí.This dissertation is focused on the development of skills of children in preschool age. The work is split into the theoretical and practical parts. The chapters of the theoretical part deal with the characteristics of child's progress in preschool age as well as with partial function development, personal growth and upbringing in the preschool age. The work is focused on the education of children in preschools and on the monitoring of school maturity. The practical part of the dissertation concentrates on the implementation of the NTC Stimulation Programme into the pedagogical and educational activities of the Salomon Preschool, a private education center for preschool children. In the practical part, the NTC method is evaluated based on the comparison of the research group with a control group consisting of children attending state preschool. Furthermore appropriate activities for preschool children are recommended. The recommended activities are focused on the support of specific key competencies and skills. KEY WORDS Partial functions, preschool age, child's development in preschool age, options of development of partial functions, school maturity, stimulation, education of children.Katedra speciální pedagogikyPedagogická fakultaFaculty of Educatio

Conductive path applied to clothing materials

katedra: KOD; přílohy: 1CD; rozsah: 67This paper describes the development of intelligent textiles, namely applying the conductive tracks on the clothing material. Its aim is to find variations in the application of conductive pathways with subsequent termination, which is applied with metal press studs. In the experimental part is a description of mechanized machinery that makes it possible to apply the fasteners in conjunction with the conductive paths of the clothing material. What methods are feasible and what is not. The state examines the requirements for the products above and no doubt deals with contemporary facilities conductive paths.Tato práce se zabývá vývojem inteligentních textilií, konkrétně aplikováním vodivých drah na oděvní materiál. Jejím cílem je snaha nalézt různé variace při aplikování vodivých drah s následným ukončením, které je aplikované pomocí kovových druků. V experimentální části je popis mechanizovaného strojního zařízení, díky kterému lze druky aplikovat ve spojení s vodivými drahami na oděvní materiál. Jaké způsoby jsou realizovatelné a jaké nikoliv. Rozebírá současný stav požadavků na výrobky výše uvedené a bezpochyby se zabývá soudobými možnostmi vodivých drah

Single-cell-based system to monitor carrier driven cellular auxin homeostasis

Background: Abundance and distribution of the plant hormone auxin play important roles in plant development. Besides other metabolic processes, various auxin carriers control the cellular level of active auxin and, hence, are major regulators of cellular auxin homeostasis. Despite the developmental importance of auxin transporters, a simple medium-to-high throughput approach to assess carrier activities is still missing. Here we show that carrier driven depletion of cellular auxin correlates with reduced nuclear auxin signaling in tobacco Bright Yellow-2 (BY-2) cell cultures. Results: We developed an easy to use transient single-cell-based system to detect carrier activity. We use the relative changes in signaling output of the auxin responsive promoter element DR5 to indirectly visualize auxin carrier activity. The feasibility of the transient approach was demonstrated by pharmacological and genetic interference with auxin signaling and transport. As a proof of concept, we provide visual evidence that the prominent auxin transport proteins PIN-FORMED (PIN) 2 and PIN5 regulate cellular auxin homeostasis at the plasma membrane and endoplasmic reticulum (ER), respectively. Our data suggest that PIN2 and PIN5 have different sensitivities to the auxin transport inhibitor 1-naphthylphthalamic acid (NPA). Also the putative PIN-LIKES (PILS) auxin carrier activity at the ER is insensitive to NPA in our system, indicating that NPA blocks intercellular, but not intracellular auxin transport. Conclusions: This single-cell-based system is a useful tool by which the activity of putative auxin carriers, such as PINs, PILS and WALLS ARE THIN1 (WAT1), can be indirectly visualized in a medium-to-high throughput manner. Moreover, our single cell system might be useful to investigate also other hormonal signaling pathways, such as cytokinin

Molekulární mechanismus transportu rostlinného hormonu auxinu do buněk

Nerovnoměrná distribuce regulátoru růstu rostlin (fytohormonu) auxinu zprostředkovává mnoho základních procesů ve vývoji rostlin. Auxin je transportován v rostlinném těle buď vaskulárním systémem nebo z buňky do buňky prostřednictvím tzv. polárního transportum auxinu (PAT). PAT zahrnuje pasivní transport auxinu do buněk difuzí kombinovaný s aktivním přenosem auxinových molekul prostřednictvím specializovaných přenašečů auxinu do buňky a z buňky. Tato práce je zaměřena na studium procesů podílejících se na přenosu auxinu do rostlinných buněk. Na základě molekulárně-biologické a biochemické charakterizace byla potvrzena funkce proteinu PaLAX1 z plané třešně Prunus avium jako přenašeče auxinu do buněk. Izolovaná cDNA genu PaLAX1 a jeho proteinové produkty jsou velmi podobné sekvencím cDNA a odpovídajících proteinových produktů genů rodiny AUX1/LAX z modelové rostliny A. thaliana, které kódují přenašeče auxinu do buněk. Na úrovni orgánů a jednotlivých buněk jsme prokázali, že nadprodukce proteinu PaLAX1 v transgenních liniích způsobuje zvýšení obsahu nativního auxinu kyseliny indol-3-octové i zvýšený příjem syntetického auxinu, kyseliny 2,4-dichlorfenoxyoctové. Dále byl zkoumán mechanismus účinku inhibitorů přenašeče auxinu do buňky kyseliny 1- naftoxyoctové (1-NOA), kyseliny 2-naftoxyoctové (2-NOA), a...The non-uniform distribution of the plant growth regulator (phytohormone) auxin is known to mediate many fundamental processes in plant development. Auxin is transported through the plant body either via vascular pathways or from cell to cell by specialized polar auxin transport machinery. This machinery consists of a balanced system of passive diffusion combined with the activities of auxin influx and efflux carriers. This work is focused on the processes that are involved in the uptake of auxin into plant cells. On the basis of molecular-biological and biochemical characterization, the function as an auxin influx carrier was confirmed for PaLAX1 protein from wild cherry (Prunus avium). The sequences of isolated cDNA of the PaLAX1 gene and of its protein product are highly similar to both the cDNAs and the corresponding protein products of the AUX1/LAX-type genes, coding for putative auxin influx carriers in model plant A. thaliana. On the level of organs and single cells, we have shown that the overproduction of PaLAX1 in transgenic lines resulted in an increase of the content of native auxin indole-3-acetic acid as well as of the uptake of synthetic auxin, 2,4-dichlorophenoxyacetic acid. Further, the mechanism of action of putative auxin influx inhibitors 1-naphthoxyacetic acid (1-NOA),...Department of BiochemistryKatedra biochemieFaculty of SciencePřírodovědecká fakult

Silver ions increase plasma membrane permeability through modulation of intracellular calcium levels in tobacco BY-2 cells

ions increase plasma membrane permeability for water and small organic compounds through their stimulatory effect on plasma membrane calcium channels, with subsequent modulation of intracellular calcium levels and ion homeostasis. The action of silver ions at the plant plasma membrane is largely connected with the inhibition of ethylene signalling thanks to the ability of silver ion to replace the copper cofactor in the ethylene receptor. A link coupling the action of silver ions and cellular auxin efflux has been suggested earlier by their possible direct interaction with auxin efflux carriers or by influencing plasma membrane permeability. Using tobacco BY-2 cells, we demonstrate here that besides a dramatic increase of efflux of synthetic auxins 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-naphthalene acetic acid (NAA), treatment with AgNO3 resulted in enhanced efflux of the cytokinin trans-zeatin (tZ) as well as the auxin structural analogues tryptophan (Trp) and benzoic acid (BA). The application of AgNO3 was accompanied by gradual water loss and plasmolysis. The observed effects were dependent on the availability of extracellular calcium ions (Ca2+) as shown by comparison of transport assays in Ca2+-rich and Ca2+-free buffers and upon treatment with inhibitors of plasma membrane Ca2+-permeable channels Al3+ and ruthenium red, both abolishing the effect of AgNO3. Confocal microscopy of Ca2+-sensitive fluorescence indicator Fluo-4FF, acetoxymethyl (AM) ester suggested that the extracellular Ca2+ availability is necessary to trigger the response to silver ions and that the intracellular Ca2+ pool alone is not sufficient for this effect. Altogether, our data suggest that in plant cells the effects of silver ions originate from the primal modification of the internal calcium levels, possibly by their interaction with Ca2+-permeable channels at the plasma membrane

Auxin influx inhibitors 1-NOA, 2-NOA, and CHPAA interfere with membrane dynamics in tobacco cells

The phytohormone auxin is transported through the plant body either via vascular pathways or from cell to cell by specialized polar transport machinery. This machinery consists of a balanced system of passive diffusion combined with the activities of auxin influx and efflux carriers. Synthetic auxins that differ in the mechanisms of their transport across the plasma membrane together with polar auxin transport inhibitors have been used in many studies on particular auxin carriers and their role in plant development. However, the exact mechanism of action of auxin efflux and influx inhibitors has not been fully elucidated. In this report, the mechanism of action of the auxin influx inhibitors (1-naphthoxyacetic acid (1-NOA), 2-naphthoxyacetic acid (2-NOA), and 3-chloro-4-hydroxyphenylacetic acid (CHPAA)) is examined by direct measurements of auxin accumulation, cellular phenotypic analysis, as well as by localization studies of Arabidopsis thaliana L. auxin carriers heterologously expressed in Nicotiana tabacum L., cv. Bright Yellow cell suspensions. The mode of action of 1-NOA, 2-NOA, and CHPAA has been shown to be linked with the dynamics of the plasma membrane. The most potent inhibitor, 1-NOA, blocked the activities of both auxin influx and efflux carriers, whereas 2-NOA and CHPAA at the same concentration preferentially inhibited auxin influx. The results suggest that these, previously unknown, activities of putative auxin influx inhibitors regulate overall auxin transport across the plasma membrane depending on the dynamics of particular membrane vesicle

Auxin transport at cellular level: new insights supported by mathematical modelling

The molecular basis of cellular auxin transport is still not fully understood. Although a number of carriers have been identified and proved to be involved in auxin transport, their regulation and possible activity of as yet unknown transporters remain unclear. Nevertheless, using single-cell-based systems it is possible to track the course of auxin accumulation inside cells and to specify and quantify some auxin transport parameters. The synthetic auxins 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthalene-1-acetic acid (NAA) are generally considered to be suitable tools for auxin transport studies because they are transported specifically via either auxin influx or efflux carriers, respectively. Our results indicate that NAA can be metabolized rapidly in tobacco BY-2 cells. The predominant metabolite has been identified as NAA glucosyl ester and it is shown that all NAA metabolites were retained inside the cells. This implies that the transport efficiency of auxin efflux transporters is higher than previously assumed. By contrast, the metabolism of 2,4-D remained fairly weak. Moreover, using data on the accumulation of 2,4-D measured in the presence of auxin transport inhibitors, it is shown that 2,4-D is also transported by efflux carriers. These results suggest that 2,4-D is a promising tool for determining both auxin influx and efflux activities. Based on the accumulation data, a mathematical model of 2,4-D transport at a single-cell level is proposed. Optimization of the model provides estimates of crucial transport parameters and, together with its validation by successfully predicting the course of 2,4-D accumulation, it confirms the consistency of the present concept of cellular auxin transport

Evolutionary conserved cysteines function as cis-acting regulators of Arabidopsis PIN-FORMED 2 distribution

Coordination of plant development requires modulation of growth responses that are under control of the phytohormone auxin. PIN-FORMED plasma membrane proteins, involved in intercellular transport of the growth regulator, are key to the transmission of such auxin signals and subject to multilevel surveillance mechanisms, including reversible post-translational modifications. Apart from well-studied PIN protein modifications, namely phosphorylation and ubiquitylation, no further post-translational modifications have been described so far. Here, we focused on root-specific Arabidopsis PIN2 and explored functional implications of two evolutionary conserved cysteines, by a combination of in silico and molecular approaches. PIN2 sequence alignments and modeling predictions indicated that both cysteines are facing the cytoplasm and therefore would be accessible to redox status-controlled modifications. Notably, mutant pin2C−A alleles retained functionality, demonstrated by their ability to almost completely rescue defects of a pin2 null allele, whereas high resolution analysis of pin2C−A localization revealed increased intracellular accumulation, and altered protein distribution within plasma membrane micro-domains. The observed effects of cysteine replacements on root growth and PIN2 localization are consistent with a model in which redox status-dependent cysteine modifications participate in the regulation of PIN2 mobility, thereby fine-tuning polar auxin transport

The IDEAL (Insulin therapy DE-intensificAtion with iglarLixi) randomised controlled trial—study design and protocol

Introduction Multiple daily injection insulin regimen (MDI) represents the most intensive insulin regimen used in the management of people with type 2 diabetes (PwT2D). Its efficacy regarding glycaemic control is counterbalanced by the increased risk of hypoglycaemia, frequently observed tendency to weight gain and necessity for frequent glucose monitoring. Recent introduction of novel antidiabetic medications with pleiotropic effects reaching far beyond the reduction of glycaemia (HbA1c), such as the glucagon-like peptide 1 receptor agonist (GLP-1 RA), has significantly widened the therapeutic options available for management of T2D. Consequently, there is currently a substantial number of PwT2D for whom the MDI regimen was initiated at a time when no other options were available. Yet, in present times, these individuals could benefit from simplified insulin regimens ideally taking advantage of the beneficial effects of the novel classes of antidiabetic medications. iGlarLixi (Suliqua®) is a once-daily fixed-ratio combination of basal insulin analogue glargine 100 U/ml and a GLP-1 RA lixisenatide. Methods Insulin therapy DE-intensificAtion with iglarLixi (IDEAL) is a six-centre, open-label, parallel-group, active comparator, phase IV randomised controlled trial with a 24-week active treatment period examining the efficacy and safety of MDI regimen de-intensification with once-daily administration of iGlarLixi versus MDI regimen continuation in PwT2D on a backgroud therapy with metformin ± sodium-glucose cotransporter 2 inhibitor. Planned Outcomes The primary objective is to compare the effects of MDI therapy de-intensification with iGlarLixi versus MDI regimen continuation regarding glycaemic control (HbA1c). Secondary objectives include detailed evaluation of the effects of MDI regimen de-intensification with iGlarLixi on glycaemic control using standardised continuous glucose monitoring (CGM) metrics and self-monitoring of plasma glucose. Furthermore, body weight and body composition analysis, quality of life and safety profile are evaluated. Trial Registration ClinicalTrials.gov, identifier NCT04945070