107 research outputs found
Mechanistic studies of substrate activation by the human CDK-activating kinase and its inhibition by small-molecule ligands
No full textThe human cyclin-dependent kinase (CDK)-activating kinase (CAK), a heterotrimeric protein complex consisting of CDK7, cyclin H, and MAT1, is a dual regulator of both transcription and the cell cycle. As a subcomplex of the general transcription factor TFIIH, it regulates transcription initiation and co-transcriptional pre-mRNA processing through phosphorylation of the C-terminal domain of RNA polymerase II. As a free complex, it regulates the cell cycle by catalysing the activating phosphorylation of CDK proteins. Due to its key activities in such fundamental cellular processes, which are commonly dysregulated in tumorigenesis, several small-molecule inhibitors of CDK7 have been developed as anti-cancer drugs.Despite a wealth of existing structural and biochemical data concerning the architecture, functions, and inhibition of the human CAK, several key questions remain. For instance, we currently lack an understanding of the molecular mechanisms by which the CAK recognises its CDK substrates. Additionally, targeting of CDK7 for cancer treatment is challenged by issues of non-selective binding of inhibitors to related CDKs, as well as possible treatment resistance.The work presented in this thesis aims to address several of these outstanding questions and challenges. I present structural and mechanistic studies of CAK-substrate complexes, which reveal the basis of CDK substrate recognition by the CAK. I additionally present structures of the CAK bound to a series of small-molecule inhibitors, thereby providing insights into inhibitor selectivity relevant to the design of future drugs, while establishing workflows for high-throughput, high-resolution structure determination by cryo-EM applicable to structure-based drug design efforts of other challenging targets. Finally, I present structural and biochemical analysis of a mutant form of the CAK that is resistant to certain inhibitors. Together, this work advances our understanding of the functions of this “master regulator” and how it can be more effectively targeted in disease
Resistance to CDK7 inhibitors directed by acquired mutation of a conserved residue in cancer cells
No full textCDK7 has emerged as a cancer target because of its pivotal roles in cell cycle progression and transcription. Several CDK7 inhibitors (CDK7i) are now in clinical evaluation. Identifying patients most likely to respond to treatment and early detection of tumour evolution towards resistance are necessary for optimal implementation of cancer therapies. Continuous culturing of prostate cancer cells with Samuraciclib, a non-covalent ATP-competitive CDK7i, led to outgrowth of resistant cells. These were characterised by the acquisition of a single base change in the CDK7 gene, Asp97 to Asn (D97N). Mutant cells were resistant to other non-covalent CDK7i but remained sensitive to covalent CDK7i. Cryo-EM structure and kinase ligand affinity determinations revealed reduced affinity of the CDK7-D97N mutant for non-covalent CDK7i. Remarkably, Asp97 is absolutely conserved in human CDKs, inferring its importance for the activities of all CDKs. Consistent with this, mutation of the homologous residue in CDK12 (D819N) or CDK4 (D99N) promoted resistance to drugs that inhibit these CDKs. Our findings reveal a general mechanism for acquired resistance with obvious implications for patients treated with CDK inhibitors
Power, policing and language policy mechanisms in schools: a response to Hudson
No full text© The Author(s) 2020. This discussion is a response to Richard Hudson's response to my article, ‘The policy and policing of language in schools’ (Cushing 2019). Hudson argues that current education policy in England generally rejects and avoids prescriptivism and sets out to illustrate this in reference to a number of policy documents. As in my original article, I conceive of language policy as p/Political and one way in which language ideologies get turned into practices, through a series of policy mechanisms such as curricula, tests, and guidance for teachers. I show how these mechanisms do not ‘reject’ prescriptivism, but explicitly perpetuate it, and thus act as a system of coercion which can lead teachers into reproducing these ideologies in their practice. I argue that Hudson's argument is limited because of its depoliticised stance and understanding of key sociolinguistic concepts and issues, such as ‘Standard English’, ‘linguistic correctness’, and language education itself. (Language education policy, language ideologies, critical applied linguistics, schools, England)
High-resolution cryo-EM of the human CDK-activating kinase for structure-based drug design
No full textAbstract Rational design of next-generation therapeutics can be facilitated by high-resolution structures of drug targets bound to small-molecule inhibitors. However, application of structure-based methods to macromolecules refractory to crystallization has been hampered by the often-limiting resolution and throughput of cryogenic electron microscopy (cryo-EM). Here, we use high-resolution cryo-EM to determine structures of the CDK-activating kinase, a master regulator of cell growth and division, in its free and nucleotide-bound states and in complex with 15 inhibitors at up to 1.8 Å resolution. Our structures provide detailed insight into inhibitor interactions and networks of water molecules in the active site of cyclin-dependent kinase 7 and provide insights into the mechanisms contributing to inhibitor selectivity, thereby providing the basis for rational design of next-generation therapeutics. These results establish a methodological framework for the use of high-resolution cryo-EM in structure-based drug design
Policy mechanisms of the standard language ideology in England’s education system
No full text© 2021 The Author(s). This article examines textual traces of the standard language ideology within current education policy in England, focusing on post-2010 reforms which are characterised by a (re)shift towards conservatism, discipline, and standards. Using tools and methods from critical stylistics and the critical discourse analysis of language policy, I interrogate a number of mechanisms which textually reinforce and reproduce the standard language ideology: curriculum documents, assessment instruments, national test materials and guidance for teachers. Whilst previous criticisms of current policy have focused on individual policy mechanisms, in this article I examine these mechanisms as a cluster, showing how they work together as de facto language policy. I show how teachers are presented with a de-historicised and de-politicised version of standardised English which masks the structural power relations that are embedded in language, and how they are constructed as standard language role-models who have a professional duty to reproduce the standard language ideology
‘Say it like the Queen’: the standard language ideology and language policy making in English primary schools
No full text© 2020 The Author(s). This article presents an analysis of the standard language ideology within a corpus of school-designed language policy documents from 264 primary schools in England. It examines the processes by which standard language ideological concepts (e.g. ‘Standard English’, ‘correctness’, ‘hegemony’) get textually manifested in school policies, and how these are intertextually and interdiscursively shaped by the broader educational policy context that teachers work in, notably the large-scale curriculum and assessment reforms of National Curriculum 2014. Using tools and methods from critical language policy, I reveal how new meanings emerge in the machinery of the policy-making process and at the contact points between policy levels. I trace how the standard language ideology within government policies gets reconstructed in school policies, with an emphasis on linguistic ‘correctness’ and the near-exclusive requirement for students and teachers to use standardised English in speech and writing. I discuss policies of surveillance, whereby teachers are discursively constructed and positioned as standard language ‘role models’: as powerful and authoritative figures who are granted a license to police, regulate and suppress their students’ language, whilst also having their own language controlled and monitored. Finally, I argue for the place of critical language awareness within the policy-making process at school level
Grammar tests, de facto policy and pedagogical coercion in England’s primary schools
No full textThe data set associated with this paper can be found at the following location: 10.17633/rd.brunel.c.4720436 Grammar tests, de facto policy and pedagogical coercion in England’s primary schools.© The Author(s) 2020. Since their introduction by the Conservative government in 2013, primary school children in England have taken a mandated grammar, punctuation and spelling assessment, which places an emphasis on decontextualised, standardised English and the identification of traditional grammatical terminology. Despite some concise criticisms from educational linguists, there remains no detailed and critical investigation into the nature of the tests, their effects on test takers, and the policy initiatives which led up to their implementation. This article contributes to this gap in knowledge, using critical language testing as a methodological framework, and drawing on a bricolage of data sources such as political speeches, policy documents, test questions and interviews with teachers. I discuss how the tests work as de facto language policy, implemented as one arm of the government’s ‘core-knowledge’ educational agenda, underpinned by a reductive conceptualisation of language and a problematic discourse of ‘right/wrong’ ways of speaking. I reveal how teachers talk about the ‘power’ of the tests, intimidating and coercing them into pedagogies they do not necessarily believe in or value, which ultimately position them as vehicles for the government’s conservative and prescriptive language ideologies
Diagnosis and management of Cushing disease
No full textjournal articleThe management of a case involving Cushing disease, defined as excess secretion of adrenocorticotropic hormone (ACTH) from a pituitary adenoma, remains one of the most vexing problems facing the pituitary surgeon. The systemic consequences of this endocrinopathy are profound morbidity and premature death in the unfortunate patients in whom remission fails to occur. For this reason, the challenge to endocrinologists and neurosurgeons charged with treating these patients is normalization of pathological ACTH and, thus, serum cortisol levels. Unfortunately, transsphenoidal surgery fails to produce remission of symptoms in a significant percentage of patients, and repeated surgery or the institution of adjuvant therapy must be considered in such cases. In this issue of Neurosurgical Focus, a series of papers has been selected to provide a comprehensive overview of the diagnosis and management of this complicated disease. Papers involving the contemporary diagnostic workup and pitfalls, as well as success rates and outcomes following transsphenoidal surgery, are presented. Author\u27s present various treatment options for patients in whom initial surgery has failed to resolve symptoms. A fascinating review paper on the phenomenon of brain atrophy seen in Cushing disease and its possible pathogenesis is included. Finally, two papers?one a historical overview and the other a contemporary review of the management of Nelson syndrome, a dreaded complication of an aggressive ACTH-secreting tumor of the pituitary developing after bilateral adrenalectomy?are presented. I would like to acknowledge the efforts of Margie Shreve and Dr. Martin Weiss. Without their expert and timely help, compiling this issue would not have been possible
A Balance of Primary and Secondary Values: Exploring a Digital Legacy
No full textThis exploratory research explores the concept of a digital legacy as a general concept and as a collection of digital possessions with unique characteristics. The results reported in this article are part of a larger study. In Cushing (2013), the author identified the characteristics of a digital possession. In this study, these characteristics of a digital possession were utilized to explore how the characteristics of several digital possessions could form a collection, or a digital legacy. In addition to being explored as a collection of digital possessions, data was collected about the general concept of a digital legacy. In part I of the study, 23 participants from three age groups were interviewed about their general concept of a digital legacy. Five general characteristics describing a digital legacy were identified. In part II of the study, interview data from Cushing (2013) was used to create statements describing digital possessions. The statements were classified utilizing the archival concept of primary and secondary values, as well as the consumer behavior concepts of self extension to possessions and possession attachment. Primary value refers to the purpose for which the item was created, while secondary value refers to an additional value that the participants can perceive the item to hold, such as a perception that an item can represent one’s identity. Using standard Q method procedure, 48 participants were directed to rank their agreement with 60 statements (written on cards), along a distribution of -5 to +5, according to the characteristics of the digital possession they would most like to maintain for a digital legacy. The ranked statements were analyzed using Q factor analysis, in order to perceive the most common statements associated with maintaining digital possessions for a digital legacy. Q method results suggested that most individuals described the digital possessions they wanted to maintain for a digital legacy using various combinations of characteristics associated with primary and secondary values. This suggests that while some participants will respond to personal archiving based on the concept of preserving identity (a perceived secondary value), this will not appeal to everyone. Information professional could consider this difference in appeal when marketing personal archiving assistance to patrons. URL: http://ijkcdt.net/xml/01030/01030.pd
The New Logic of Work: How Automation Is Changing Public Sector Practice
No full textRecent advancements in automation technologies such as Robotic Process Automation (RPA) and Artificial Intelligence (AI) have made it possible to automate repetitive administrative tasks, offering the promise of increased efficiency and improved service delivery. These technologies are increasingly being integrated into the daily operations of public organizations. Despite the growing academic interest, limited research has focused on how RPA/AI integration reshapes everyday administrative work practices. This thesis goes some way in filling this knowledge gap by exploring how work unfolds within a Swedish municipality that in recent years has implemented RPA/AI across the organization. This thesis adopts a work-as-practice perspective and applies a qualitative research method, with data collected through semi-structured interviews. The findings show that on an operational level, the integration of RPA/AI led to a shift in performed tasks, from manual data entry to supervisory and exception-handling roles, resulting in freed-up time that was reallocated to professional judgement. On an individual level, work practices were reshaped by changes in how employees engage with their roles, developed competence, and interacted with technology in their daily routines. This shift enabled greater involvement across the organization and altered how employees understood their roles and relationships with clients. At the organizational level, role distributions evolved, job titles shifted, and teams became more cross-functional. Employees came to view their work in a wider organizational context, which facilitated internal knowledge transfer and contributed to a more open and forward-thinking work culture. This thesis demonstrates that RPA/AI are not simply technical tools introduced to optimize workflows, but they actas mediators of how work is carried out, understood and structured. The research contributes to existing knowledge by offering a contextualized understanding of the integration of RPA/AI in the public sector.Nya framsteg inom automationsteknologier, såsom Robotiserad processautomation (RPA) och Artificiell intelligens (AI), har möjliggjort automatisering av repetitiva administrativa uppgifter med löftet om ökad effektivitet och förbättrad tjänsteleverans. Dessa teknologier integreras i allt större utsträckning i den dagliga verksamheten hos offentliga organisationer. Trots ett växande akademiskt intresse, finns det begränsad forskning om hur integrationen av RPA/AI omformar det dagliga administrativa arbetet. Denna uppsats bidrar till att fylla detta akademiska gap genom att undersöka hur arbete utformar sig inom en svensk kommun som under de senaste åren har implementerat RPA/AI inom organisationen. Uppsatsen utgår från ett arbetspraktiskt perspektiv och använder en kvalitativ forskningsmetod, där data samlats in genom semistrukturerade intervjuer. Resultaten visar att på en operativ nivå ledde integrationen av RPA/AI till en förskjutning i arbetsuppgifter från manuell datainmatning till övervakning och hantering av undantag, vilket frigjorde tid som i stället kunde ägnas åt professionella bedömningar. På individnivå omformades arbetspraktiker genom förändringar i hur medarbetare engagerade sig i sina roller, utvecklade kompetens och samspelade med teknologin i sina dagliga rutiner. Denna förändring möjliggjorde ett ökat engagemang inom organisationen och påverkade hur medarbetare förstod sina roller och relationer till klienter. På organisatorisk nivå utvecklades nya rollfördelningar, yrkestitlar förändrades och team blev mer tvärfunktionella. Medarbetare började se sitt arbete i ett bredare organisatoriskt sammanhang, vilket underlättade intern kunskapsöverföring och bidrog till en mer öppen och framåttänkande arbetskultur. Denna uppsats visar att RPA/AI inte enbart är tekniska verktyg för att optimera arbetsflöden, utan även fungerar som förmedlare av hur arbete utförs, förstås och struktureras. Forskningen bidrar till den existerande litteraturen genom att erbjuda en kontextualiserad förståelse för integrationen av RPA/AI inom offentlig sektor
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