Study Purpose and Rationale The United States is witnessing growth in its aging population and an increase in the prevalence of end-stage renal disease, resulting in a substantial rise in kidney transplantation among adults age 65 and over. While older adult recipients fare better clinically than their counterparts on dialysis, far less is known about their psychosocial wellbeing following transplantation. A small body of qualitative work in organ transplantation suggests that there is an important period of adaptation following transplantation during which a recipient must incorporate the new organ into his or her sense of identity and bodily integrity. Difficulty navigating the adaptation process can have significant adverse effects on psychosocial and clinical outcomes, in extreme cases resulting in graft loss and even death. Employing a phenomenological approach, this study sought to better understand the lived experience of older adult kidney transplant recipients and the impact of kidney transplantation on their sense of identity and bodily integrity, with implications for the design of effective supports and interventions to optimize post-transplant mental health and wellbeing. Methods Applying the hermeneutic phenomenology of philosopher Paul Ricoeur, this study explored the lived experience of 10 kidney transplant recipients age 65 and over. Participants completed one to two in-depth phenomenological interviews lasting approximately one hour each. Interview transcripts and field notes were analyzed using an inductive approach, beginning with an epoché (bracketing) phase and followed by the processes of phenomenological reduction (textured description), imaginative variation (structured description) and synthesis to arrive at an understanding or approximation of the universal essence of the phenomenon of adapting to a kidney transplant. Findings Participants reported experiencing the “strangeness” of incorporating the organ of another person into their own body and concern over specific corporeal changes associated with dialysis and transplantation, expressing a disruption of their ‘idem’ sense of identity that ranged from mildly unsettling to deeply distressing. However, participants also articulated powerful narratives of resilience and coping, describing their ability to overcome adversity as an anchor of their ‘ipse’ sense of identity that allowed them to navigate idem corporeal changes over time. The continuity and strength of participants’ ipse sense of self over the life course enabled them to adjust to profound corporeal change, and for some participants kidney transplantation represented a form of liberation from ‘machine life’ (dialysis) and restoration of their idem sense of self. Conclusions/Implications Findings from this study have significant implications for ensuring the provision of optimal support to older adult kidney transplant recipients. A more nuanced understanding of the post-transplant experience of adapting to a new organ will enhance the informed consent process, providing the opportunity for transplant teams, potential recipients and their caregivers to engage in meaningful dialogue about the risks and benefits of the procedure and helping to establish expectations for post-transplant recovery. By shedding light on older adult recipients’ perspectives, findings also will help to inform the design of future research studies addressing candidacy for transplantation and post-transplant quality of life. Furthermore, study results will assist transplant teams in determining how to optimize post-transplant care and support for older adult recipients, thereby improving outcomes and contributing to the overall wellbeing of this population. Finally, the findings have implications for ongoing policy discussions in the field, including how to ensure equitable patient selection by establishing appropriate age-related criteria for kidney transplant recipients.

Bakgrund: Antalet njurtransplantationer har under de senaste åren ökat i Sverige. För patienter med CKD-5 är det den säkraste åtgärden för att kunna leva ett längre och bekvämare liv. En njurtransplantation från en levande donator bidrar till bättre återhämtning och längre liv. Den grundutbildade sjuksköterskan behöver ha god insikt i ämnet njurtransplantation för att ge god och säker vård, både medicinskt och omvårdnadsmässigt. Syfte: Syftet med denna litteraturstudie är att utforska patienters upplevelser i väntan på njurtransplantation med en levande donator. Metod: I litteraturstudien används tio vetenskapliga artiklar med kvalitativ metod.  Resultat: Resultatet belyser två huvudteman, förväntningar i väntan samt förändrade relationer. Deltagarna i studien upplevde känslor som hopp, oro, förhoppning om att kunna återgå till ett normalt liv samt skuld. Mottagarnas upplevelser varierade beroende på vilken relation de hade till donatorn. Slutsats: Det är viktigt för den grundutbildade sjuksköterskan att ge korrekt omvårdnad och god information till patienten. Sjuksköterskan kan bidra till god omvårdnad genom stöttande samtal och att initiera kontakt mellan donator och mottagare. Mottagarna upplever levande njurtransplantation som livsomvälvande, och många olika känslor och funderingar väcks till liv. / Background: The number of kidney transplants has increased in Sweden in recent years. Kidney transplantation is the safest measure for patients living with CKD-5, it prolongs and leads to a better life. A kidney donation from a living donor contributes to better recovery and a longer life. The graduate nurse needs good insight regarding the subject of kidney transplantation to be able to provide adequate and safe care, both in the aspect of medicine and care. Aim: The aim of this literature study is to explore patients' experiences while waiting for a kidney transplantation from a living donor.  Method: In this literary study ten scientific articles were used with a qualitative method. Results: In the result two main themes arose, Expectations in waiting and Changing relations. The participants of this study experienced feelings of hope, worry, hope of getting back to a normal life and guilt. Recipients' experiences varied depending on what relation they had to the donor. Conclusion: It is of importance to the graduate nurse to provide just care and correct information to the patient. The nurse is able to contribute to just care through supportive conversation and initiating contact between the donor and the recipient. The recipient perceives waiting for kidney transplantation as life changing, and many diverse feelings appear.

Researchers have identified several methods for treating acute myocardial infarction (AMI) patients affected by ischemia and reperfusion injury. Some of these therapies include thrombolysis, balloon angioplasty, and coronary arterial bypass graft (CAGB). This lab has previously demonstrated that transplantation of mitochondria into the ischemic zone of a rabbit heart during reperfusion significantly improved recovery as compared to current techniques. In order for this therapy to be translated into the clinic a rapid isolation method for producing highly pure and functional mitochondria will be required. Previously described mitochondrial isolation methods using differential centrifugation and/or Ficoll gradient centrifugation require 60 to 100 minutes to complete. Herein, a method for rapid isolation of mitochondria from mammalian tissue biopsies is described. In this protocol, manual homogenization is replaced with the tissue dissociator's standardized homogenization cycle. This allows for uniform and consistent homogenization of tissue that is not easily achieved with manual homogenization. Following tissue dissociation, the homogenate is filtered through nylon mesh filters which eliminates repetitive centrifugation steps. Mitochondrial isolation time is less than 30 minutes compared to 60-100 minutes using alternative methods. This isolation protocol yields approximately 2 x 10^10 viable and respiration competent mitochondria from 0.18 ± 0.04 g (wet weight) tissue sample.

Call number: LD2668 .T4 1962 V47

Transplantation is the treatment of choice for end stage organ failure however current immunosuppressive therapies whilst effective at preventing acute allograft rejection, fail to prevent late graft loss due to chronic rejection and are associated with an increased risk of infection and malignancy. Therefore there is a clear unmet clinical need for improved strategies to prevent allograft rejection. OX40 is a member of the TNFR superfamily that has potent costimulatory properties. Although the impact of blockade of the OX40-OX40L pathway has been well documented in models of autoimmune disease, its effect on the rejection of allografts is less well defined. Therefore the aim of this thesis was to determine the impact of OX40 blockade on conventional and regulatory T cell responses to allografts. We found that activation of CD4+ and CD8+ naïve and memory T cells resulted in the induction of OX40 expression and that blockade of OX40-OX40L interactions partially inhibited the response of alloreactive T cells in vitro and prevented skin allograft rejection but did not result in the induction of tolerance. OX40 blockade was found to have no effect on the activation and proliferation of T cells but rather effector T cells failed to accumulate and migrate to skin allografts. This was shown to be the result of an enhanced degree of cell death amongst proliferating effector cells. In addition, blockade of OX40-OX40L interactions at a time of exposure to alloantigen resulted in a pool of Treg with an enhanced ability to suppress T cell responses to alloantigen in vitro and in vivo. Counter-intuitively, OX40 blockade was found to increase the potency of alloreactive Treg by promoting survival following re-activation. Finally, although OX40 blockade impacted both conventional and regulatory T cell responses, anti-OX40 administration did not promote skin or heart allograft survival in immunocompetent recipients and failed to synergise with blockade of other costimulatory molecules to prevent allograft rejection. In conclusion, these data demonstrate that blockade of OX40-OX40L interactions can attenuate naïve and memory T cell responses to alloantigen whilst promoting the survival of alloreactive Treg. Therefore, we propose that anti-OX40 would be a worthwhile adjunct to pre-existing strategies to induce tolerance.

Mucopolysaccharidosis type IIIA (MPS IIIA) is caused by mutations in the N-sulphoglucosamine sulphohydrolase (SGSH) gene, leading to cellular accumulation of heparan sulphate and progressive neurodegeneration in patients. One of the proposed treatment methods is haematopoietic stem cell (HSC) gene therapy, which should result in an excess of SGSH produced in the peripheral organs and brain. The pre-clinical feasibility of this approach was demonstrated by our group in a mouse model of MPS IIIA. However, the overall efficiency of this method was limited and a number of approaches to solving these issues were addressed in this project in order to bring this therapy closer to clinical application. Our first aim was to optimise transduction of HSCs using cytokines, bovine serum albumin (BSA), and chemicals, such as MG132, genistein and valproic acid. Addition of BSA with cytokines improved cell viability, addition of MG132/ BSA/ cytokines improved transduction, but also caused cellular toxicity, while addition of genistein was inefficient. Addition of valproic acid with cytokines resulted in increased number of colony forming units. Next, we generated clinically applicable third generation pCCL lentiviral vector backbones with the eGFP reporter gene driven by one of ubiquitous hPGK or myeloid specific hCD11b and hCD18 internal human promoters, and optimised production of lentiviral vectors to increase titre and reduce production cost. These lentiviral vectors were used to transduce lineage depleted HSCs and transplanted into WT mice. Full chimerism and over 80% transduction were achieved with an average of 5 vector copy numbers/ cell. The hCD11b promoter resulted in the highest eGFP expression in monocytes and B cells in blood, but was weaker than the hPGK in T cells. The hCD18 promoter was more monocyte-specific but weak. Significant numbers of GFP-positive microglial cells were present in the brain from all groups, with an average of 25% transduced CD11b-positive cells in perfused mice. We subsequently codon-optimised (CO) the SGSH gene significantly improving enzyme activity, and transduced lineage depleted WT cells with one of hCD18.SGSH-CO, hCD11b.SGSH-CO, or hPGK.SGSH-CO lentiviral vectors, or MPS IIIA cells with either hCD11b.SGSH-CO or hPGK.SGSH-CO lentiviral vectors. These transduced cells were transplanted into MPS IIIA mice and outcomes were measured 6 months later. Only treatment with the hCD11b.SGSH-CO-LV transduced WT or MPS IIIA HSCs corrected abnormal behaviour of MPS IIIA mice. However, all treatments resulted in complete GAG storage clearance in the periphery and brain, and significantly elevated enzyme activity in the brain, liver and spleen to 7-11%, 60-75%, and 170-250% of WT enzyme activity respectively. A fine threshold of over 8.6% brain enzyme activity appeared to be required for behavioural correction in MPS IIIA mice. Further assessment of treated mice for the amount of secondary storage, HS sulphation patterning, neuroinflammation and longevity are still required for complete therapeutic assessment. However, it appears that neurological correction of the MPS IIIA mouse using MPS IIIA cells is feasible using a clinically-relevant pCCL vector with the hCD11b promoter and the codon-optimised SGSH gene.

Spinal root avulsion leads to paralysis and loss of sensory function. Surgical methods can improve motor function and ameliorate pain but sensory recovery in adults is poor. Previous studies have shown that cell transplantation or treatment with trophic factors can improve functional outcome in rodents after dorsal root transection or crush. Here, a dorsal root injury model, more similar to human avulsion injuries, was used. The aims of this thesis were to investigate the behaviour of different stem cells following transplantation to avulsed dorsal roots and asses their potential to serve as possible regenerative therapy. In paper I, different murine stem cell types were transplanted to avulsed dorsal roots in rats. Murine embryonic stem cells remained outside the spinal cord and were surrounded by glutamatergic terminals. Boundary cap neural crest stem cells (bNCSC) formed elongated bands outside the spinal cord and migrated to the spinal cord as single cells. In paper II, transplanted bNCSC were further characterized. bNCSC remaining outside the spinal cord expressed glial markers and were associated with different types of sensory fibres. bNCSC that migrated into the injured spinal cord expressed different neuronal markers. In paper III, effects of bNCSC transplantation on local vasculature and glial scar formation were studied. bNCSC increase angiogenesis in a non dose response manner and participate in boundary glial scar formation. In paper IV, bNCSC spinal migration was analysed using two different injury models - dorsal root transection and dorsal root avulsion. In addition, bNCSC capacity to support sensory regeneration was assessed and the results suggest that bNCSC do not support robust regeneration of avulsed afferents. In paper V, an in vitro stem cell model system was used to assess the possibility of using artificial nanomaterials to deliver differentiation factors. Cells treated with either soluble factors or particle-delivered factors showed similar differentiation patterns. Stem cell transplantation offers several opportunities following dorsal root avulsion, including cell replacement and regenerative support. By elucidating the mechanisms by which stem cells can assist regeneration of avulsed afferents will allow for more targeted or combinatorial approaches, including growth factor treatment.

published_or_final_version / Surgery / Doctoral / Doctor of Philosophy

published_or_final_version / Anatomy / Doctoral / Doctor of Philosophy

published_or_final_version / Medicine / Master / Master of Philosophy