Constructive Empathizing – Educational Competence in the Light of Child’s Play

Pre-school play is an important moment in a child’s development.  Caretakers play an important role during play. This article presents a structure of educational competence called “constructive empathizing” or “invitingaccompanying competence”. A caretaker’s special task during child’s play is conversion between the third-person knowledge and a subjective, autotelic interaction. The caretaker’s constructive empathizing during a child’s play determines the child’s social and moral development, it helps the child to turn play into work, as a sign of higher spontaneous understanding of cultural norms standing behind cultural correctness actions. Finally, it is evidence of dynamic interaction between abstract-concrete thinking, with the child learning from the caretaker

Many facets of CPEB proteins in neurons and beyond: expression, mRNA recognition and phosphorylation

CPEBs are a family of evolutionary conserved auxiliary translational factors. They bind to CPEs in the 3’UTR of target mRNA and, by interacting with other members of the translational machinery, promote or decrease gene expression. Since the first description of CPEB protein as a key orchestrator of oocyte maturation in Xenopus Laevis, key functions of CPEBs in embryonic development, cellular senescence and synaptic plasticity have been described. Local, postsynaptic modulation of synaptic efficacy is the most studied function of CPEBs in the CNS. However, the expression of CPEBs is not restricted to neurons and even 7% of total brain mRNAs possesses CPEs. Moreover, specific mRNAs are enriched in functionally distinct subcellular regions, like dendritic spines of neurons or complex cellular processes of astrocytes, where they might be translationally regulated. Herein, I embarked on further elucidating the role of CPEB translational regulators family in selected cell populations in mouse brain. In the first part of the work, the target mRNA specificity of different CPEBs was tested. I argued with the hypothesis that CPEBs -3 and -4 require mRNA secondary structure (a stem-loop) for target recognition. I showed that despite differences in the RBD primary structure, CPEB-1 shows a considerable sequence specificity overlap with CPEB-3 (and possibly, due to sequence similarity, with CPEBs -2 and -4 as well). This holds true for neuronal (CaMKIIα) and astrocytic(β-catenin) CPEB targets. Additionally, RNAco-immunoprecipitation revealed two novel CPEB target mRNAs, encoding (1) an astrocytic gap junction protein (Connexin43) and (2) CPEB-3 protein itself. In the first part of the work, the target mRNA specificity of different CPEBs was tested. I argued with the hypothesis that CPEBs -3 and -4 require mRNA secondary structure (a stem-loop) for target recognition. I showed that despite differences in the RBD primary structure, CPEB-1 shows a considerable sequence specificity overlap with CPEB-3 (and possibly, due to sequence similarity, with CPEBs -2 and -4 as well). This holds true for neuronal (CaMKIIα) and astrocytic(β-catenin) CPEB targets. Additionally, RNAco-immunoprecipitation revealed two novel CPEB target mRNAs, encoding (1) an astrocytic gap junction protein (Connexin43) and (2) CPEB-3 protein itself. In the second part of the thesis, CPEBs 1-4 gene expression in microglia, NG2 cells and astrocytes was assessed at the mRNA and protein level, confirming the ubiquitous nature of the CPEB expression. A more detailed analysis of CPEB-3 mRNA revealed expression of only certain isoforms of the protein in ESdM cells. Each CPEB family member occurs in several splice variants. Hitherto, only a handful of reports are dealing with the significance of alternative splicing of CPEBs. Interestingly, regulation of expression of CPEBs in neurons may happen in an isoform-specific fashion. Seeing this as a highly intriguing phenomenon, I focused on CPEB-3, containing a putative kinase recognition site on the alternatively spliced exon 5. In a cell-free systems and in cultured cells, I showed that CPEB-3 is a target of PKA and CaMKII – both critical for LTP/LTD, and that exon 5 harbors a consensus sequence required for kinase recognition. In vivo, in a transgenic mouse overexpressing CPEB3 protein in principal neurons of the hippocampus, I founda significant decrease in GluR2 AMPA-R subunit protein levels. In view of the above, the stimulation-induced phosphorylation of CPEB-3 may very well be a novel mechanism of translational modulation of synaptic plasticity. In the final part of the thesis, I aimed at establishing a quality control method for Cre-mediated gene deletions. Using Cx30-/-;Cx43fl/fl mice with hGFAP-Cre recombinase driver, I developed a reliable immunoblot-based tool for post-hoc control of recombination variability in hGFAP-Cre transgenic mice

Distinct translatome changes in specific neural populations precede electroencephalographic changes in prion-infected mice

Selective vulnerability is an enigmatic feature of neurodegenerative diseases (NDs), whereby a widely expressed protein causes lesions in specific cell types and brain regions. Using the RiboTag method in mice, translational responses of five neural subtypes to acquired prion disease (PrD) were measured. Pre-onset and disease onset timepoints were chosen based on longitudinal electroencephalography (EEG) that revealed a gradual increase in theta power between 10- and 18-weeks after prion injection, resembling a clinical feature of human PrD. At disease onset, marked by significantly increased theta power and histopathological lesions, mice had pronounced translatome changes in all five cell types despite appearing normal. Remarkably, at a pre-onset stage, prior to EEG and neuropathological changes, we found that 1) translatomes of astrocytes indicated reduced synthesis of ribosomal and mitochondrial components, 2) glutamatergic neurons showed increased expression of cytoskeletal genes, and 3) GABAergic neurons revealed reduced expression of circadian rhythm genes. These data demonstrate that early translatome responses to neurodegeneration emerge prior to conventional markers of disease and are cell type-specific. Therapeutic strategies may need to target multiple pathways in specific populations of cells, early in disease

Instrumental Value of Knowledge and Knowledge-creation

John Dewey’s pragmatic theory of knowledge supports the assertion that both the instrumental value of knowledge and knowledge-creation have common philosophical underpinnings. An analysis of this issue can help bridge the gap between the typical economic approach to knowledge and those approaches that are represented by contemporary epistemology and philosophy of science. The universal concept of the value of knowledge that is proposed here allows one to carry out further research on using contemporary epistemology in knowledge management practice.3249643Studia Metodologiczn

Evaluation of effectiveness of waterjet propulsor for a small underwater vehicle

The goal of the project described is to replace the existing propulsion system of a small underwater vehicle with a solution less prone to mechanical damage and ensuring a lower risk of the entanglement of fibrous objects suspended in the body of water. Four typical marine screws are utilised in the current design of the vehicle. One possible solution of the problem is the application of waterjet propulsors located inside the body of the vehicle instead. The general conditio of the application of the new solution was to secure at least the same motion control capabilities of the vehicle while the basic capability is its propulsion effectiveness at the required speed. Specific features of the considered waterjet propulsor, when compared with their application in surface vessel propulsion, are the lack of the head losses and the low significance of cavitation issues. One of the difficulties in the considered case is the small diameter of the propulsor in comparison to commercially available waterjet units, which have diameters between 0.1 [m] and 1.0 [m]. There is very little data regarding the design and performance of devices in the 0.02 to 0.05 [m] range. Methods utilised to forecast the performance of the new propulsion system are presented and results compared. These were semi-empirical calculations, numerical calculations and tests of real devices. The algorithm that is based on semi-empirical calculations is of particular interest while it offers possibility quick assessment of performance of a propulsor composed of several well defined components. The results indicate the feasibility of modification of the propulsion system for the considered vehicle if all the existing circumstances are taken into account

A Study of Acid-Base Equilibria in Acetonitrile Systems of 2-Halo(Cl,Br,I)-4-nitropicoline(3,5,6) N-oxides

An attempt has been made to determine potentiometrically (1) acid dissociation constants of cations obtained by protonation of nine trisubstituted pyridine N-oxides, namely 2-halo(Cl, Br and I)-4-nitropicoline N-oxides with the methyl group at positions 3, 5, and 6, as well as (2) the cationic homoconjugation constants of these cationic acids with conjugated N-oxides in acetonitrile. On the basis of the substitution effect, variations of the acid dissociation constants of the trisubstituted pyridine N-oxide cations are discussed. The determined pKa values of the protonated 2-halo-4-nitropicoline N-oxides are compared with the previously determined equilibrium constants of the cationic acids conjugated with the mono- and disubstituted pyridine N-oxides in acetonitrile. Further, based on the pKa values of the protonated 2-halo-4-nitropicoline N-oxides in acetonitrile, supplemented with correlations between pKa’s of the protonated mono- and disubstituted pyridine N-oxides in acetonitrile and water, the pKa\u27s of the acids conjugated with the trisubstituted N-oxides studied in aqueous solutions have been estimated. Moreover, it has been concluded that the cationic homoconjugation constants cannot be determined by potentiometric titration in acetonitrile solutions of the 2-halo-4-nitropicoline N-oxide systems

Manipulating the Prion Protein Gene Sequence and Expression Levels with CRISPR/Cas9

The mammalian prion protein (PrP, encoded by Prnp) is most infamous for its central role in prion diseases, invariably fatal neurodegenerative diseases affecting humans, food animals, and animals in the wild. However, PrP is also hypothesized to be an important receptor for toxic protein conformers in Alzheimer's disease, and is associated with other clinically relevant processes such as cancer and stroke. Thus, key insights into important clinical areas, as well as into understanding PrP functions in normal physiology, can be obtained from studying transgenic mouse models and cell culture systems. However, the Prnp locus is difficult to manipulate by homologous recombination, making modifications of the endogenous locus rarely attempted. Fortunately in recent years genome engineering technologies, like TALENs or CRISPR/Cas9 (CC9), have brought exceptional new possibilities for manipulating Prnp. Herein, we present our observations made during systematic experiments with the CC9 system targeting the endogenous mouse Prnp locus, to either modify sequences or to boost PrP expression using CC9-based synergistic activation mediators (SAMs). It is our hope that this information will aid and encourage researchers to implement gene-targeting techniques into their research program