N-terminal myristoylation is required for membrane localization of cGMP-dependent protein kinase type II

The apical membrane of intestinal epithelial cells harbors a unique isozyme of cGMP-dependent protein kinase (cGK type II) which acts as a key regulator of ion transport systems, including the cystic fibrosis transmembrane conductance regulator (CFTR)-chloride channel. To explore the mechanism of cGK II membrane-anchoring, recombinant cGK II was expressed stably in HEK 293 cells or transiently in COS-1 cells. In both cell lines, cGK II was found predominantly in the particulate fraction. Immunoprecipitation of solubilized cGK II did not reveal any other tightly associated proteins, suggesting a membrane binding motif within cGK II itself. The primary structure of cGK II is devoid of hydrophobic transmembrane domains; cGK II does, however, contain a penultimate glycine, a potential acceptor for a myristoyl moiety. Metabolic labeling showed that cGK II was indeed able to incorporate [3H]myristate. Moreover, incubation of cGK II-expressing 293 cells with the myristoylation inhibitor 2-hydroxymyristic acid (1 mM) significantly increased the proportion of cGK II in the cytosol from 10 +/- 5 to 35 +/- 4%. Furthermore, a nonmyristoylated cGK II Gly2 --> Ala mutant was localized predominantly in the cytosol after transient expression in COS-1 cells. The absence of the myristoyl group did not affect the specific enzyme activity or the Ka for cGMP and only slightly enhanced the thermal stability of cGK II. These results indicate that N-terminal myristoylation fulfills a crucial role in directing cGK II to the membrane

Translocation of BCR to chromosome 9: A new cytogenetic variant detected by FISH in two Ph-negative, BCR-positive patients with chronic myeloid leukemia

Leukemic cells from two patients with Philadelphia-negative chronic myeloid leukemia (CML) were investigated: I) Cytogenetics showed a normal 46.XY karyotype in both cases, 2) molecular studies revealed rearrangement of the M-BCR region and formation of BCR-ABL fusion mRNA with b2a2 (patient I) or b3a2 (patient 2) configuration, and 3) fluorescence in situ hybridization (FISH) demonstrated relocation of the 5′ BCR sequences from one chromosome 22 to one chromosome 9. The ABL probe hybridized to both chromosomes 9 at band q34, while two other probes which map centromeric and telomeric of BCR on 22q 11 hybridized solely with chromosome 22. For the first time, a BCR-ABL rearrangement is shown to take place on 9q34 instead of in the usual location on 22q 11. A rearrangement in the latter site is found in all Ph-positive CML and in almost all investigated CML with variant Ph or Ph-negative, BCR-positive cases. The few aberrant chromosomal localizations of BCR-ABL recombinant genes found previously were apparently the result of complex and successive changes. Furthermore in patient 2, both chromosomes 9 showed positive FISH signals with both ABL and BCR probes. Restriction fragment length polymorphism (RFLP) analysis indicated that mitotic recombination had occurred on the long arm of chromosome 9 and that the rearranged chromosome 9 was of paternal origin. The leukemic cells of this patient showed a duplication of the BCR-ABL gene, analogous to duplication of the Ph chromosome in classic CML. In addition they had lost the maternal alleles of the 9q34 chromosomal region. The lymphocytes of patient 2 carried the maternal chromosome 9 alleles and were Ph-negative as evidenced by RFLP and FISH analyses, respectively. © 1993 Wiley-Liss, Inc

When disruptive technologies knock at the door: talking about ChatGPT in higher education - A qualitative case study investigating the perceptions at Radboud University about ChatGPT in higher education.

When ChatGPT was released in November 2022, the world of higher education was overwhelmed by the advanced, versatile applications of the language model that seemed both exiting and threatening. Nevertheless, few research on the perceptions of Higher Education Institutions (HEI) regarding ChatGPT existed. Therefore, utilizing a qualitative case method, the present study aimed at acquiring further understanding of these attitudes towards the language model in higher education. In three in-depth semi-structured interviews with employees from the Radboud University in the Netherlands, the following research question was explored: “How does Radboud University perceive the use of ChatGPT in higher education?” A thematic analysis indicated that participants expected the disruptive technology to alter conventional learning and teaching at Radboud University. However, while some accepted the presence of ChatGPT, others warned for embracing ChatGPT too quickly. Furthermore, the ubiquity of ChatGPT increased awareness of the importance of academic competences. Besides, the focus on a conscious attitude was considered vital when utilizing ChatGPT. Although participants recognized beneficial applications for higher education, the system behind ChatGPT, and its commercial character were alarming. Yet, the formulation of adequate university measures may be difficult, as ChatGPT’s capabilities continuingly evolve. Conclusively, the perceptions about ChatGPT in higher education identified at the Radboud University may contribute to ongoing discussions within HEI on how to deal with the prevalence of ChatGPT and its implications for academic education

LA TEORIA KANTIANA DELLA MOTIVAZIONE MORALE NELLA 'FONDAZIONE DELLA METAFISICA DEI COSTUMI'. UN'ANALISI STORICO-CRITICA

The Thesis \uabLa teoria kantiana della motivazione morale nella Fondazione della metafisica dei costumi. Un\u2019analisi storico-critica\ubb aims to reconstruct Kant\u2019s theory of moral motivation as it is formulated in Groundwork of the Metaphysics of Morals. This Ph.D. Thesis is predominantly a commentary on the passages from Kant\u2019s Groundwork that are relevant to understanding his account of moral motivation. At the same time, as a background to the work, it considers the whole Kantian corpus with its differing theories of moral motivation. The Thesis includes four chapters, each devoted to the analysis of one of four conceptual areas of the Groundwork\u2019s theory of moral motivation: (1) practical reason, will and their reciprocal relationship, (2) will and good will, (3) the feeling and Triebfeder of respect, and (4) the Highest Good. The contemporary debate on Kant\u2019s moral motivation focuses on two opposite interpretations of it: viz., the affectivist and the intellectualist one. In order to choose between these interpretations, the letter of the Kantian texts must be thoroughly analyzed and their theoretical plausibility must be assessed. In the Introduction and in the first chapter I study Kant\u2019s theory of what a human action is and I give an interpretation of Kant\u2019s use of the adverb unmittelbar, which leads me to view Kant\u2019s theory of moral motivation as an affective one. In the remainder of the work I give further grounds to this interpretation by means of a detailed analysis of the text supported by a contextual theoretical study. Furthermore, the first chapter includes (a) the definition of practical reason and will as functions of the human mind, (b) the consequent identification of the laws according to which a rational being has the power of acting as the linking element between these functions and the human faculty of reason and (c) the examination of the secondary literature\u2019s positions about which kind of laws are the ones of human actions. I also provide an answer to the latter question, specifically addressing the issue of distinguishing the formal structure of these laws from their normative validity. The second chapter\u2019s first purpose is to understand the extent of philosophical and psychological possible knowledge about will and good will. To achieve this, I study the concept of moral experience as it appears in the Critique of Practical Reason. The second chapter also investigates Lust and Unlust, since I argue that they deeply mark Kantian conception of both sensible perception and moral motivation. Finally, in the chapter\u2019s conclusion I analyze the teleological argument of the Groundwork\u2019s first section, its structure, its role in the entire work, and the relevance it assumes by taking into account the systematic function of the Kantian notion of character. The third chapter presents a study on the different meanings that Kant\u2019s notion of respect assumes throughout the Groundwork, in the second Critique and in the Doctrine of Virtue. This analysis focuses in particular on the Kantian terms Bewegungsgrund and Triebfeder. The conclusion I draw from it is an assessment of Kant\u2019s peculiar form of a priori affectivism. The final chapter\u2019s arguments stem from the conclusions I drew in Chapter two concerning the relevance of teleology with regard to Kantian theory of morality and moral motivation. Besides commenting on the two explicitly numbered propositions about duty in Section one of the Groundwork (GMS, AA 04: 399.35-400.03 and GMS, AA 04: 400.17-19), I identify the first one, which has not been explicitly mentioned by Kant. The first proposition states the inclusion of the concept of good will in that of duty (GMS, AA 04: 397.06-08). This leads to the analysis of the meaning of Kant\u2019s definition of good will as good in itself, and, at the same time, not corresponding to the whole and Highest Good. I finally study the alleged inconsistency and irrelevancy of Kant\u2019s conception of the Highest Good in the framework of his theory of moral motivation, and I refute it: I eventually prove exactly the opposite, i.e. I point out the great importance and the coherent role that the Highest Good plays in Kant\u2019s theory of moral motivation