The role of p53 and CYLD in mitochondrial death pathways and mechanisms of neuronal necroptosis

Neuronal cell death causes progressive loss of brain tissue and function after acute brain injury and in chronic neurodegenerative diseases. Although the pathological features of stroke and brain trauma or Alzheimer’s and Parkinson’s disease differ greatly, the underlying neuronal damage shares common molecular and cellular mechanisms. Despite extensive research and increasing knowledge on the molecular pathology, no efficient therapy has been born from these efforts until today. As a promising concept to overcome this plight, it has been suggested to enhance endogenous survival signaling pathways like the transcription factor NF-κB and thus obtain neuroprotection. Increasing neuroprotective NF-κB signaling can be achieved by blocking repressors of NF-κB transcriptional activity such as p53 and CYLD. Both factors may mediate cell death by mechanisms dependent on and independent of NF-ΚB signaling. Therefore, the major aim of this study was to explore the roles of p53 and CYLD in neuronal cell death and to connect their detrimental effects with NF-κB activity. This issue was addressed in immortalized mouse hippocampal HT-22 neurons and in primary neuronal cultures exposed to glutamate toxicity. Furthermore, an in vivo model system of traumatic brain injury was employed to compare infarct development after controlled cortical impact in wild-type and CYLD-/- mice. The present study revealed that both approaches, inhibiting p53 and CYLD successfully preserved mitochondrial integrity and function, and significantly attenuated neuronal cell death. Surprisingly, however, the pronounced neuroprotective effect of the p53-inhibitor pifithrin-α occurred independently of enhanced NF-κB activity in HT-22 cells. In addition, neuroprotection induced by silencing of CYLD was completely independent of NF-κB, despite of the previously established role of CYLD as a negative regulator of NF-κB in keratinocytes. In line with that notion, the NF-κB subunit expression and NF-κB transcriptional activity were not significantly altered in HT-22 neurons undergoing glutamate dependent cell death. In conclusion, these data suggested that the NF-κB pathway was neither significantly affected by glutamate dependent cell death, nor did it mediate the neuroprotective response of CYLD and p53 inhibition in this model system of glutamate toxicity. Interestingly, inhibiting p53 with pifithrin-α maintained mitochondrial morphology and mitochondrial membrane potential in HT-22 cells. This effect occurred independently of p53 dependent transcription. Investigating the underlying cause of neuroprotection associated with CYLD depletion, it was unveiled that glutamate-induced oxytosis in HT-22 cells occurred through mechanisms of necroptosis. This conclusion is based on the detection of RIP1/RIP3 complexes as a hallmark of necroptotic cell death in HT-22 cells exposed to glutamate. Further, silencing either RIP-kinase provided strong protection of the cells. Repressing CYLD, in turn, prevented the formation of the RIP1/RIP3 necrosome, suggesting that inhibition of necroptosis was the underlying mechanism of neuroprotection after CYLD depletion. In contrast, CYLD depletion had no effect on cell death in a model of glutamate excitotoxicity in primary cultured neurons, while inhibition of RIP1 kinase by necrostatin-1 significantly enhanced neuronal survival. These data suggest a CYLD independent but RIP1 dependent mechanism of glutamate toxicity in primary neurons, which requires further investigation. In vivo, however, using a model of traumatic brain injury, CYLD knockout mice showed a significantly reduced infarct size compared to wild-type littermates suggesting a potent neuroprotective effect inherent with CYLD repression. In summary the data from this thesis highlight a yet unknown role of CYLD in neuronal cell death and unravel CYLD and p53-dependent mechanisms of cell death as a putative therapeutic approach for the treatment of acute and chronic neurodegenerative diseases. Future research, however, is warranted to further elucidate the exact mechanisms leading to CYLD and RIP-kinase activation in neurons and to determine the exact molecular link to mitochondria

A World Remade: Graham Greene\u27s Thrillers And The Nineteen-thirties

Within Graham Greene\u27s large body of work stand several texts which, for reasons explored in this thesis, he originally called entertainments . Because this label seems to suggest that these texts are not as important as his other novels, they have received relatively little critical attention. This thesis helps to redress this imbalance.;Beginning with a brief consideration of generic distinction, I argue that Greene\u27s use of the entertainment label is tied to the specific historical, political, and literary context of the nineteen-thirties in Britain. At this time, Greene and other writers reacted to the literary and critical practices of the high modernists, who emerged during and after World War One. With a renewed sense that literature could not be divorced from the social and political milieu of which it was a part, Greene and others sympathetic to the cause of the Left returned to a realistic mode of fiction and used popular forms of writing in their works. Greene\u27s particular response, seen primarily in his entertainments , was to develop the classical detective story as practiced by Poe, Doyle, Christie, and others into the political thriller.;By looking at the form of the detective story along lines suggested by Roland Barthes, Tzvetan Todorov, Peter Brooks, and Dennis Porter, I show how Greene\u27s texts explore the problem of reading and understanding in an intensely political age. In this context, his texts are seen as narrative (s) of narratives (Brooks 25).;After discussing several of Greene\u27s early texts, such as Murder for the Wrong Reason, Rumour at Nightfall, Stamboul Train, It\u27s a Battlefield, and England Made Me, in terms of how they develop a political critique while exploring aspects of popular literature, I turn to a consideration of A Gun for Sale, Brighton Rock, The Confidential Agent, and The Ministry of Fear. In my examination of these texts, I consider a number of related issues involving the production and interpretation of narrative, and I relate these concerns to questions of ideology in politics and literary criticism. Ultimately, I find The Ministry of Fear to be Greene\u27s fullest treatment of the materials of detective fiction

Collaborating with Businesses to Support and Sustain Research

Financial assistance is necessary for sustaining research at universities. Business collaborations are a potential means for obtaining these funds. To secure funding, understanding the process for obtaining these business funds is important for nursing faculty members. Although faculty rarely request funding from businesses, they are often in a position to solicit financial support due to existing relationships with clinical agency administrators, staff, and community leaders. The economic support received from businesses provides outcomes in nursing research, research education, academic–service partnerships, and client health care. This article describes the steps and processes involved in successfully obtaining research funding from businesses. In addition, case examples for securing and maintaining funding from health care agencies (evidence-based practice services) and from a health manufacturing company (product evaluation) are used to demonstrate the process

Jones v. Chagrin Falls: Muddying the Statutory Waters of Ohio\u27s Administrative Law Appeal Process

The common-law doctrine of failure to exhaust administrative remedies has generally been held to be a prerequisite to judicial review in statutorily defined administrative law appeal processes. Similarly, the United States Supreme Court in interpreting the federal administrative law appeal process, and the case law on Ohio\u27s administrative law appeal process, have found that the doctrine of exhaustion is a jurisdictional bar to a declaratory judgment action except while challenging the constitutionality of a municipal or administrative decision. However, according to the holding in Jones v. Chagrin Falls, this may no longer be the case in Ohio. This article discusses the Jones case in detail, starting with the factual and procedural history in Parts II, and then moving on to the court’s opinion and rationale in Part III. The article finishes up with the author’s overall analysis of the decision in Part IV

Jones v. Chagrin Falls: Muddying the Statutory Waters of Ohio\u27s Administrative Law Appeal Process

The common-law doctrine of failure to exhaust administrative remedies has generally been held to be a prerequisite to judicial review in statutorily defined administrative law appeal processes. Similarly, the United States Supreme Court in interpreting the federal administrative law appeal process, and the case law on Ohio\u27s administrative law appeal process, have found that the doctrine of exhaustion is a jurisdictional bar to a declaratory judgment action except while challenging the constitutionality of a municipal or administrative decision. However, according to the holding in Jones v. Chagrin Falls, this may no longer be the case in Ohio. This article discusses the Jones case in detail, starting with the factual and procedural history in Parts II, and then moving on to the court’s opinion and rationale in Part III. The article finishes up with the author’s overall analysis of the decision in Part IV

Can Large Language Models assist in Hazard Analysis?

Large Language Models (LLMs), such as GPT-3, have demonstrated remarkable natural language processing and generation capabilities and have been applied to a variety tasks, such as source code generation. This paper explores the potential of integrating LLMs in the hazard analysis for safety-critical systems, a process which we refer to as co-hazard analysis (CoHA). In CoHA, a human analyst interacts with an LLM via a context-aware chat session and uses the responses to support elicitation of possible hazard causes. In this experiment, we explore CoHA with three increasingly complex versions of a simple system, using Open AI's ChatGPT service. The quality of ChatGPT's responses were systematically assessed to determine the feasibility of CoHA given the current state of LLM technology. The results suggest that LLMs may be useful for supporting human analysts performing hazard analysis

Metal Complexes of Phosphinic Acids: Part XIII*-Dithiophosphinic Acid Complexes of UO2(VI) & In(III)

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Incremental Assurance Through Eliminative Argumentation

An assurance case for a critical system is valid for that system at a particular point in time, such as when the system is delivered to a certification authority for review. The argument is structured around evidence that exists at that point in time. However, modern assurance cases are rarely one-off exercises. More information might become available (e.g., field data) that could strengthen (or weaken) the validity of the case. This paper proposes the notion of incremental assurance wherein the assurance case structure includes both the currently available evidence and a plan for incrementally increasing confidence in the system as additional or higher quality evidence becomes available. Such evidence is needed to further reduce doubts engineers or reviewers might have. This paper formalizes the idea of incremental assurance through an argumentation pattern. The concept of incremental assurance is demonstrated by applying the pattern to part of a safety assurance case for an air traffic control system