Mechanism of damage sensing and cell killing following the inhibition of nucleotide excision repair by fludarabine in quiescent cells

Inhibition of DNA repair by the nucleoside of fludarabine (F-ara-A) induces toxicity in quiescent human cells. The sensing and signaling mechanisms following DNA repair inhibition by F-ara-A are unknown. The central hypothesis of this project was that the mechanistic interaction of a DNA repair initiating agent and a nucleoside analog initiates an apoptotic signal in quiescent cells. The purpose of this research was to identify the sensing and signaling mechanism(s) that respond to DNA repair inhibition by F-ara-A. Lymphocytes were treated with F-ara-A, to accumulate the active triphosphate metabolite and subsequently DNA repair was activated by UV irradiation. Pre-incubation of lymphocytes with 3 μM F-ara-A inhibited DNA repair initiated by 2 J/m2 UV and induced greater than additive apoptosis after 24 h. Blocking the incorporation of F-ara-A nucleotide into repairing DNA using 30 μM aphidicolin considerably lowered the apoptotic response. ^ Wild-type quiescent cells showed a significant loss in viability than did cells lacking functional sensor kinase DNA-PKcs or p53 as measured by colony formation assays. The functional status of ATM did not appear to affect the apoptotic outcome. Immunoprecipitation studies showed an interaction between the catalytic sub-unit of DNA-PK and p53 following DNA repair inhibition. Confocal fluorescence microscopy studies have indicated the localization pattern of p53, DNA-PK and γ-H2AX in the nucleus following DNA damage. Foci formation by γ-H2AX was seen as an early event that is followed by interaction with DNA-PKcs. p53 serine-15 phosphorylation and accumulation were detected 2 h after treatment. Fas/Fas ligand expression increased significantly after repair inhibition and was dependent on the functional status of p53. Blocking the interaction between Fas and Fas ligand by neutralizing antibodies significantly rescued the apoptotic fraction of cells. ^ Collectively, these results suggest that incorporation of the nucleoside analog into repair patches is critical for cytotoxicity and that the DNA damage, while being sensed by DNA-PK, may induce apoptosis by a p53-mediated signaling mechanism. Based on the results, a model is proposed for the sensing of F-ara-A-induced DNA damage that includes γ-H2AX, DNA-PKcs, and p53. Targeting the cellular DNA repair mechanism can be a potential means of producing cytotoxicity in a quiescent population of neoplastic cells. These results also provide mechanistic support for the success of nucleoside analogs with cyclophosphamide or other agents that initiate excision repair processes, in the clinic. ^

The natural killer gene complex genetic locus Chok encodes Ly-49D, a target recognition receptor that activates natural killing

Previously, we established that natural killer (NK) cells from C57BL/6 (B6), but not BALB/c, mice lysed Chinese hamster ovary (CHO) cells, and we mapped the locus that determines this differential CHO-killing capacity to the NK gene complex on chromosome 6. The localization of Chok in the NK gene complex suggested that it may encode either an activating or an inhibitory receptor. Here, results from a lectin-facilitated lysis assay predicted that Chok is an activating B6 NK receptor. Therefore, we immunized BALB/c mice with NK cells from BALB.B6–Cmv1r congenic mice and generated a mAb, designated 4E4, that blocked B6-mediated CHO lysis. mAb 4E4 also redirected lysis of Daudi targets, indicating its reactivity with an activating NK cell receptor. Furthermore, only the 4E4+ B6 NK cell subset mediated CHO killing, and this lysis was abrogated by preincubation with mAb 4E4. Flow cytometric analysis indicated that mAb 4E4 specifically reacts with Ly-49D but not Ly-49A, B, C, E, G, H, or I transfectants. Finally, gene transfer of Ly-49DB6 into BALB/c NK cells conferred cytotoxic capacity against CHO cells, thus establishing that the Ly-49D receptor is sufficient to activate NK cells to lyse this target. Hence, Ly-49D is the Chok gene product and is a mouse NK cell receptor capable of directly triggering natural killing.

Yvonne Wanrow on trial for killing the man who molested her child

General note: Title and date provided by Bettye Lane.

The Corporate Manslaughter and Corporate Homicide Act 2007 or the Health and Safety (Offences) Act 2008: corporate killing and the law

This thesis examines the regulatory and legislative approach taken in the United Kingdom to deal with deaths arising from work related activities and, in particular, deaths that can be directly attributed to the behaviour of corporations and other organisations. Workplace health and safety has traditionally been seen in the United Kingdom as a regulatory function which can be traced to the very earliest days of the Industrial Revolution. With an emphasis on preventing workplace accidents and ill-health through guidance, advice and support, the health and safety legislation and enforcement regime which had evolved over the best part of two centuries was considered inadequate to effectively punish corporations considered responsible for deaths caused by their activities following a series of disasters in the late twentieth and early twenty-first centuries. To address this apparent inadequacy, the Corporate Manslaughter and Corporate Homicide Act 2007 was introduced creating the offence of corporate manslaughter and corporate homicide. Based on a gross breach of a relevant duty of care resulting in the death of a person, the Act effectively changed what had previously considered a matter of regulation, an approach that had obvious weaknesses and shortcomings, to one of crime and criminal law. Whether this is the best approach to dealing with deaths caused by an organisation is challenged in this thesis and the apparent distinction between ‘criminal’ and ‘regulatory’ offences is also examined. It was found that an amended Health and Safety at Work etc. Act 1974 to include a specific offence of corporate killing, in conjunction with the Health and Safety (Offences) Act 2008 would almost certainly have resulted in a more effective approach to dealing with organisations responsible for causing deaths as consequence of their activities. It was also found that there was no substantive difference between ‘regulatory’ and ‘criminal’ law other than the stigma associated with the latter, and that distinction would almost certainly disappear, at least in the context of worker safety, as a consequence of the penalties available following the introduction of the Health and Safety (Offences) Act 2008.

Understanding nonsense : breathing life into shibboleths and killing critical thought in higher education

In this paper, I focus on the growing "nonsense industry" which is most apparent in the writing typical of business, government departments, and the financial press. This writing, like technical writing, is characterised by heavy reliance on grammatical metaphor. It endows shibboleths - for instance, "globalisation"; "efficiencies"; "competition"; "modernisation"; "consumer sentiment"; "reform"; and so on - with anthropomorphic qualities. These anthropomorphic artefacts of technocratised language are then presented as having immutable powers over people. Thus they become banal public excuses for negligent practices in both business and government.

When does pain and distress relief hastening death become killing?

This paper discusses the question of when pain and distress relief known to hasten death would cross the line between permissible conduct and killing. The issue is discussed in the context of organ donation after cardiac death, and considers the administration of analgesics, sedatives, and the controversial use of paralysing agents in the provision and withdrawal of ventilation.

Misericordiae Antemortem - The 1955 Mount Superbus Crash

On the 9th April 1955, RAAF Lincoln Bomber A73-64, on a mercy flight to transfer a critically ill infant from Townsville to Brisbane, crashed at Mount Superbus killing the four crew and two civilians on board. The immediate search and rescue was organised by a group of Brisbane bushwalkers who were camping in the area. Police and RAAF personnel subsequently joined the civilians at the crash site to recover the victims. During their initial search of the crash they located what were believed to be the remains of five adults. The arrival of the RAAF Senior Medical Officer (SMO) the following day revealed that only four adult bodies had been found and the bodies of both civilians, an adult and infant, were missing. Later that day the remains of six victims were recovered from the crash site and conveyed to the Warwick Police Station for identification. The RAAF SMO was responsible for the identifications of the aircrew while the Government Medical Officer, police and coroner were responsible for the identifications of the civilians. Eight days later, further remains of the infant were found by a civilian looking through the wreckage. This paper uses archival records not previously researched from a Disaster Victim Identification (DVI) perspective to stimulate interest among forensic practitioners, criminologists and other interested parties in the history of DVI and how practices in Australia have evolved.

Strolling the coastline: criminology in everyday life : through ‘landscape’ from gaol to ‘badlands’

Part travelogue, part flight of fancy, this paper recounts a coastline stroll from Maroubra Beach to Bondi in Sydney’s eastern suburbs. The author as ‘travel guide’ points out features of potential interest to two visiting criminological colleagues as they ‘pass by’ scenery of great beauty shadowed by acts of spectacular violence. The everyday acts of walking and talking while passing through a ‘landscape’ serve to constitute a criminology of everyday life, illustrating the way in which a consciousness of crime, crime sites, analyses and theories permeates the ways a ‘tourist trail’ might be experienced and seen, myths made and histories forged. The walk starts with the unseen lines of penal force radiating from Long Bay Gaol, before skirting through surfing and its regulation; the ‘brotherhood’ of the BRA Boys; the Hines killing and the politics of self defence; the shark arm case, the Virgin Mary and the Bali bombing memorial at Coogee; zones of the beach and Jock Young’s Vertigo at Bronte and Tamarama; before finishing at the Marks Park ‘badlands’ at Bondi, scene of a series of mostly unsolved and unpunished homophobic killings, giving rise to reflections on ‘ungrievable lives’, memory, mourning and forgetting.

Does withdrawing life-sustaining treatment cause death or allow the patient to die?

The paper discusses the view of Franklin Miller and Robert Truog that withdrawing life-sustaining treatment causes death and so is a form of killing. I reject that view. I argue that even if we think there is no morally relevant difference between allowing a patient to die and killing her (itself a controversial view), it does not follow that allowing to die is a form of killing. I then argue that withdrawing life-sustaining treatment is properly classified as allowing the patient to die rather than as killing her. Once this is accepted, the law cannot be criticised for inconsistency by holding, as it does, that it is lawful to withdraw life-sustaining treatment but unlawful to give patients a lethal injection.

Flood victim’s heartbreak : ‘don’t be complacent about the risk’

"No one had higher stakes in the findings of the Queensland Floods Commission of Inquiry than Grantham father Matthew Keep, whose mother, mother-in-law and baby daughter, Jessica, died at Grantham that terrible day in January. For seven months he has read every statement, submission, running log, disaster management plan and media article available. As he has comforted his grieving wife, Stacy, helped care for his two young children Madison, 5, and Jacob, 4, who amazingly survived the flood, and welcomed a new baby into the family, Matthew has searched for answers for himself and his community. Why were authorities not able to warn people in seven towns in Toowoomba and the Lockyer Valley that the worst flash flooding in at least a century was about to strike, killing 22 people? How could such a sudden and catastrophic flood claim three members of his family within minutes?"