Navigating the Empty Spaces of Care: A Feminist Political Economy Analysis of the Care Experiences and Work Practices of Women Living with Cancer

Advances in cancer treatment are improving survival rates and, in so doing, transforming the nature of cancer from an acute to chronic illness. Within the Canadian health care system, there has been increasing policy acknowledgment of and support for a culture change in care that promotes a stronger chronic care agenda and yet the Canadian health care system continues to operate within a predominantly medical model that favours acute care and treatment. The Princess Margaret Cancer Centre has shown some evidence of their commitment to a chronic care model through the implementation of ELLICSR Health, Wellness and Cancer Survivorship Centre. While this appears promising, understanding how these changes are operationalized in a predominantly acute health care setting remains an underdeveloped area and the implications for patients remain unknown. In this study, I apply critical ethnography and various data collection methods (document analysis, participant observation, semi-structured interviews and photo elicitation) in order to explore the patient experience within this changing health care milieu, paying particular attention to patients care experiences and work practices. As informed and framed by feminist political economy, this project explores the everyday care and work experiences of women diagnosed with cancer as situated within the broader social, political, and economic contexts in which cancer care and work are anchored. Analysis traces key tensions and conflicts between policy directions and the everyday environments where care takes place. The findings illuminate that, in the absence of suitable and sustained institutional and funding support, ideological changes that appear to be in line with improved patient autonomy and control (e.g., person-centeredness, patient involvement and self-management) more closely resemble increased individual responsibility and work for which little choice is given. In the empty spaces between policy promises and care practices, the findings reveal a new responsibility and accountability circuit wherein access to good quality care increasingly rests on patient action/inaction, thus rendering opportunities for care more inequitable. The delivery of psychosocial and supportive care through ELLICSR made a critical difference in the care experiences of study participants; however, the precarity of this space demonstrates the lack of commitment to the proposed goals of reforms. As our health care system changes, we must take up a context-sensitive approach that invites engagement with the messiness and complexity of cancer care as conceptualized, practiced, and lived

Developing a System for the Application of Reverse Genetics to Bunyaviruses

For many positive strand RNA viruses, genomic RNA can initiate productive infection in the absence of preformed viral proteins when introduced into susceptible cells. RNA transcribed in vitro from cDNA copies of positive strand virus genomes is also infectious. This property has enabled the powerful techniques of recombinant DNA technology to be applied to produce viruses carrying defined mutations by manipulation of cDNA clones. In contrast, deproteinised genomes of negative strand RNA viruses are non infectious. The study of negative strand viruses has therefore been hampered by the absence of methodology enabling the manipulation of their genomes. The non infectious nature of negative strand genomes is due to the requirement of viral proteins to transcribe translation competent positive strand mRNA. The template for transcription by the RNA dependent RNA polymerases of negative strand viruses is a ribonucleoprotein complex (RNP). Production of infectious RNA from cDNA of negative strand RNA virus genomes is therefore thought to require correct assembly of the synthetic RNA into RNP which can then be expressed and replicated by viral RNA polymerase. This thesis describes work carried out with the overall aim of establishing a system to produce Bunyamwera virus, a segmented negative strand RNA virus, carrying defined nucleotide alterations to its genome. Towards this goal, two broad approaches were investigated to attempt to present replicating virus with synthetic RNA in a form in which it could be recognised as a transcription template by the viral polymerase. The first approach involved transcribing viral-like RNA in vivo from transfected cDNA constructs in the presence of replicating Bunyamwera virus in the hope that the intracellular pool of viral proteins would enable encapsidation and replication of the synthetic RNA. The second approach involved attempts to reconstitute RNP in vitro by incubating synthetic RNA with nucleocapsid protein derived from Bunyamwera virus. For in vivo transcription of bunyavirus-like RNA from transfected plasmids, a recombinant vaccinia virus (vTF7-3) which expresses T7 RNA polymerase was investigated to assess its suitability for producing a Bunyamwera vims small (S) segment RNA from transfected cDNA in the presence of a replicating helper bunyavirus. The helper virus used was an existing reassortant virus (Bun/Bun/Mag) containing Bunyamwera virus large (L) and middle (M) genome segments and a Maguari vims S segment. Successful replication of the plasmid derived Bunyamwera S segment by the Bun/Bun/Mag helper virus would result in a proportion of progeny virus having all three gene segments of Bunyamwera virus. Dual infection experiments were performed to investigate the compatibility of the component viruses of the rescue strategy. Metabolic labelling and Northern analysis suggested that the general strategy of rescuing a Bunyamwera virus S segment RNA, transcribed in vivo by vTF7-3, using the Bun/Bun/Mag reassortant virus, was feasible with regard to interactions of the component viruses. Transcription of Bun S segment RNA in vivo from transfected cDNA by T7 RNA polymerase supplied by vTF7-3 was investigated by Northern blot analysis. It was not possible to detect RNA of the expected size. Transcription did occur since N protein could be detected in vTF7-3 infected cells transfected with a cDNA construct designed to produce positive strand S segment RNA from a T7 promoter. In an attempt to rescue plasmid derived Bunyamwera virus S segment RNA into a Bun/Bun/Mag reassortant virus, supernatants were retained from cells which had been infected with vTF7-3 and the Bun/Bun/Mag reassortant virus and transfected with Bunyamwera virus S segment cDNA. 200 individual plaques of progeny vims were screened by metabolic labelling to determine the origin of their N protein. No Bun/Bun/Bun virus was recovered indicating that if rescue occured, it did so at a frequency of <1/200. As an alternative to screening larger numbers of progeny virus, a counter- selectable rescue virus was sought to enable selection of any progeny virus containing plasmid derived (wild type) S segment RNA. Temperature sensitive (ts) mutants of Maguari vims were further characterised in attempt to identify a vims with a S segment lesion. To enable the S segments of mutants from each of the three reassortment groups to be sequenced, a rapid PCR-based procedure was developed which allowed full-length S segment cDNA to be amplified in a single step from RNA isolated from crude preparations of virus. Using this method, full-length S segment cDNAs were amplified and cloned from a representative of each of the three reassortment groups(ts6,ts17 and ts23) and from wild type Maguari virus. Three cDNA clones of each segment were sequenced by the Sanger di-deoxy chain termination method. A single point mutation was found in the S segment cDNA of Maguari virusts23. This virus was used as a helper virus in attempt to replicate RNA transcribed in vivo from a Bunyamwera virus S segment cDNA by the vTF7-3 expression system. (Abstract shortened by ProQuest.)

Donor outcomes in anonymous live liver donation

BackgroundDeath rates on liver transplant waiting lists range from 5%-25%. Herein, we report a unique experience with 50 anonymous persons who volunteered to address this gap by offering to donate part of their liver to a recipient with whom they had no biological connection or prior relationship (A-LLD).MethodsCandidates were screened to confirm excellent physical, mental, social, and financial health. Demographics and surgical outcomes were analyzed. Qualitative interviews after donation examined motivation and experiences. Validated self-reported questionnaires assessed personality traits and psychological impact.Results50 A-LLD liver transplants (LT) were performed between 2005 and 2017. Most donors had a university education, a middle-class income, and a history of prior altruism. Half were women. Median age was 38.5 years (range 20-59 yrs.). Thirty-three (70%) learned about this opportunity through public or social media. Saving a life, helping others, generativity, and reciprocity for past generosity were motivators. Social, financial, healthcare, and legal supports in Canada were identified as facilitators. A-LLD identified most with the personality traits of agreeableness and conscientiousness. The median hospital stay was six days. There was one Dindo-Clavien Grade 3 complication that completely resolved. One-year recipient survival was 91% in 22 adults and 97% in 28 children. No A-LLD reported regretting their decision.ConclusionsThis is the first and only report of the motivations and facilitators of A-LLD in a large cohort. With rigorous protocols, outcomes are excellent. A-LLD has significant potential to reduce the gap between transplant organ demand and availability

Genomic analysis of codon, sequence and structural conservation with selective biochemical-structure mapping reveals highly conserved and dynamic structures in rotavirus RNAs with potential cis-acting functions

Rotaviruses are a major cause of acute, often fatal, gastroenteritis in infants and young children world-wide. Virions contain an 11 segment double-stranded RNA genome. Little is known about the cis-acting sequences and structural elements of the viral RNAs. Using a database of 1621 full-length sequences of mammalian group A rotavirus RNA segments, we evaluated the codon, sequence and RNA structural conservation of the complete genome. Codon conservation regions were found in eight ORFs, suggesting the presence of functional RNA elements. Using ConStruct and RNAz programmes, we identified conserved secondary structures in the positive-sense RNAs including long-range interactions (LRIs) at the 5′ and 3′ terminal regions of all segments. In RNA9, two mutually exclusive structures were observed suggesting a switch mechanism between a conserved terminal LRI and an independent 3′ stem–loop structure. In RNA6, a conserved stem–loop was found in a region previously reported to have translation enhancement activity. Biochemical structural analysis of RNA11 confirmed the presence of terminal LRIs and two internal helices with high codon and sequence conservation. These extensive in silico and in vitro analyses provide evidence of the conservation, complexity, multi-functionality and dynamics of rotavirus RNA structures which likely influence RNA replication, translation and genome packaging

A qualitative investigation of lived experiences of long-term health condition management with people who are food insecure.

Background: As more people are living with one or more chronic health conditions, supporting patients to become activated, self-managers of their conditions has become a key health policy focus both in the UK and internationally. There is also growing evidence in the UK that those with long term health conditions have an increased risk of being food insecure. While international evidence indicates that food insecurity adversely affects individual's health condition management capability, little is known about how those so affected manage their condition(s) in this context. An investigation of lived experience of health condition management was undertaken with food insecure people living in north east Scotland. The study aimed to explore the challenges facing food insecure people in terms of, i. their self-care condition management practices, and ii. disclosing and discussing the experience of managing their condition with a health care professional, and iii. Notions of the support they might wish to receive from them. Methods: Twenty in-depth interviews were conducted with individuals attending a food bank and food pantry in north east Scotland. Interview audio recordings were fully transcribed and thematically analysed. Results: Individuals reporting multiple physical and mental health conditions, took part in the study. Four main themes were identified i.e.: 1. food practices, trade-offs and compromises, that relate to economic constraints and lack of choice; 2. illness experiences and food as they relate to physical and mental ill-health; 3. (in) visibility of participants' economic vulnerability within health care consultations; and 4. perceptions and expectations of the health care system. Conclusions: This study, the first of its kind in the UK, indicated that participants' health condition management aspirations were undermined by the experience of food insecurity, and that their health care consultations in were, on the whole, devoid of discussions of those challenges. As such, the study indicated practical and ethical implications for health care policy, practice and research associated with the risk of intervention-generated health inequalities that were suggested by this study. Better understanding is needed about the impact of household food insecurity on existing ill health, wellbeing and health care use across the UK

The influenza virus panhandle is involved in the initiation of transcription.

The role of the influenza A virus panhandle structure formed from the 3'- and 5'-terminal nucleotides of virion RNA segments was studied in both an RNA polymerase binding assay and an in vitro transcription assay. Despite recent indications that promoter activity is simply a function of the 3'-terminal sequence of virion RNA, our results show that both 3'- and 5'-terminal sequences are involved in the initiation of transcription. We propose a new model for the initiation of transcription which has implications for the mechanisms by which influenza virus transcription, replication, and polyadenylation may be regulated in the infected cell

Characterization of the RNA-fork model of virion RNA in the initiation of transcription in influenza A virus.

It has been shown that both 3' and 5' conserved termini of influenza A virus virion RNA are involved in the initiation of transcription. An RNA-fork model has been proposed, according to which there is a crucial double-stranded region formed by complementary bases at positions 10 to 12 of the 3' terminus and bases at positions 11' to 13' of the 5' terminus, which are extended by 2 or 3 segment-specific base pairs. The two termini at positions 1 to 9 and 1' to 10' in the 3' and 5' termini, respectively, are in a single-stranded conformation. Here we further characterize this model, focusing on the individual roles of the proposed duplex region and the proposed two single-stranded ends. Residues within the conserved 5' terminus that are involved in the initiation of transcription were determined. Single, double, and triple mutations in the proposed duplex region provided further evidence that, for the initiation of transcription in vitro, the duplex RNA is more important than the actual sequence of these residues, although some restrictions in sequence were apparent. On the other hand, there was evidence that base pairing is not required at residues 1 to 7. We propose that the 5' terminus of virion RNA should be treated as an integral part of the virion RNA promoter and discuss a possible mechanism for the recognition of the virion RNA promoter by the influenza A virus RNA polymerase complex

In vitro transcription and polymerase binding studies of the termini of influenza A virus cRNA: evidence for a cRNA panhandle.

An in vitro transcription assay was used to study transcription from synthetic RNA corresponding to the 3' terminus of influenza A virus cRNA. Micrococcal nuclease-treated influenza virus ribonucleoprotein was used as a source of active polymerase complex. Mutations at two regions of the 13 nucleotide-long conserved cRNA 3' terminus were shown to reduce transcription templated by the short added model RNAs. The first region, at positions 1 and 2 from the 3' terminus, was shown to be affected by the exact nature of the dinucleotide primer used in the in vitro transcription reactions and may not be relevant in vivo. The second region, centred on positions 11 and 12, may be involved in base pairing with conserved nucleotides at the 5' terminus of the cRNA. Evidence for this comes from the finding that RNA corresponding to 5' conserved sequences, but mutated to restore the postulated base pairing with the mutated 3' ends, could partly restore transcription. Binding of the influenza virus polymerase complex to a set of 5'-mutated RNAs was investigated using a photochemical cross-linking assay. Specific binding to two regions of the cRNA 5' terminus was demonstrated, at positions 1 to 3 and positions 8 to 10. Together, these observations suggest that a panhandle forms from the termini of the cRNA molecule and that this structure may play a role in transcription to produce virion RNA