Consumption of fruits and vegetables has been shown to reduce the risk of various types of cancer. Macro- and micro-nutrients as well as non-nutritive phytochemicals present in fruits and vegetables have been associated with this effect. This study was conducted to investigate the chemopreventive and chemotherapeutic potential of two types of fruits which are endemic to Borneo Island: Mangifera pajang (bambangan) and Artocarpus odoratissimus (tarap). The first part of the project was to study the antioxidant potential of the crude extracts of the plants in vitro. The fruits were first separated into flesh, kernel and peel for M. pajang and flesh and seed for A. odoratissmus. DPPH (2,2- diphenyl-I-picrylhydrazyl) free radical scavenging and FRAP (ferric reducing / antioxidant power) assay were employed for the antioxidant study. The result showed that the kernel of M. pajang extract displayed strongest antioxidant activity as assessed using both assays, followed by M. pajang peel, A. odoratissmus seed, M. pajang flesh and A. odoratissmus flesh. The presence of selected phytochemicals in the plant extracts was determined in the next chapter. Polyphenols have been identified as major phytochemicals in the plant extracts, and in M. pajang kernel extract represents about 10% of its total weight. Gallic acid, coumaric acid, sinapic acid, caffeic acid, ferulic acid, chlorogenic acid, naringin, hesperidin, rutin, luteolin and diosmin have been identified as the key polyphenol phytochemicals present in the kernel of M. pajang which might be responsible for the superior antioxidant properties as compared to other extracts. Concern that the results for the chemical antioxidant assay do not necessarily reflect cellular activity led to the third part of the project; assessment of the cytoprotective activity of the crude extracts against oxidative stress induced by tert-butyl hydroperoxide (t-BHP). Only M. pajang kernel extract as well as the positive control (quercetin) displayed cytoprotective activity against this toxicant. It seems that non-cell based antioxidant assay does not necessarily reflect the activity in cell-based antioxidant assay. This is shown by lack of cytoprotective activity of both M. pajang peel and A. odoratissimus extracts despite their considerably high antioxidant activity in DPPH free radical scavenging and FRAP assay. In order to study which proteins might be involved in the cytoprotection mechanism, western blotting method was employed to determine the expression of various Cytoprotective proteins [i.e. quinone reductase (NQO I), glutathione peroxidase (OR), methionine sulfoxide reductase A (MSRA), heat shock protein 27 (HSP27) and heat shock protein 70 (HSP 70)]. Different cytoprotective mechanisms were observed by the kernel extracts and quercetin. In the present study, NQOI, OR, MSRA, HSP27 and HSP 70 have been shown to be involved in the cytoprotection activity of quercetin while only OR and MSRA were involved in the cytoprotection activity of M. pajang kernel extracts. Other cytoprotective proteins remain to be studied to fully understand the cytoprotection mechanism of both plant extract and quercetin. Some chemopreventive agents have been shown to suppress cancer proliferation, induce apoptosis in cancer cells as well as inhibit angiogenesis and metastasis in pre-clinical and clinical trials. Thus, the last part of the project was to determine the anti-cancer potential of plant extracts in a variety of cancer cell lines (derived from breast, colon, liver and ovarian carcinoma). The results showed that the kernel extract of M. pajang displayed strong anti proliferative activity in breast cancer cell lines (MCF-7 and MDA-MB-231). The kernel extract induced cell cycle arrest in MCF-7 cells at the sub-G1 (apoptosis) phase of the cell cycle in a time-dependent manner. For MDA-MB-231 cells, the kernel extract induced strong G2-M arrest in cell cycle progression at 24 hours, resulting in substantial sub-G1 (apoptosis) arrest after 48 and 72 hours of incubation. Staining with Annexin V -FITC and propidium iodide revealed that this apoptosis occurred early in both cell types, 36 hours for MCF-7 cells and 24 hours for MDA-MB- 231 cells, with 14.0% and 16.5% of the cells respectively undergoing apoptosis at these times. This apoptosis appeared to be dependent on caspases-2 and -3 in MCF-7 cells and on caspases-2, -3 and -9 in MDA-MB-231 cells. As a conclusion, from the two plants (M. pajang and A. odoratissimus) studied, the extract of M. pajang kernel displayed diverse health benefit properties, antioxidant, chemoprevention and chemotherapeutic potential. M. pajang could be fully utilized for pharmaceutical, nutraceutical as well as food products. Further study (i.e. animal and clinical study, isolation of pure compounds, bioavailability study) are required to determine the efficacy in human population.

This thesis focuses specifically on the transmission of bad news from doctors to patients and their families in the context of a hospital oncology department. It uses awareness context theory as a basis for exploring communication between patients and health care professionals, particularly when the information to be disclosed is sensitive and will have a significant bearing on how people perceive their future. In order to enhance clinical practice, senior health care professionals in particular, have in the past been encouraged through government policy and professional legislation to attend communication skills courses to develop the way they communicate and interact with patients. Yet, in spite of these interventions evidence suggests that doctors and other health care professionals still find it difficult to negotiate sensitive and emotionally challenging discussions, and frequently question whether or not patients are aware and understand the information disclosed to them and whether or not information provided meets the needs and expectations of patients. The premise of this research is that more attention needs to be given to how other more reflective and experiential professional development approaches and techniques might help doctors communicate better with their patients when disclosing sensitive information and bad news. In order to do this however, a better understanding is needed about what is going on in consultations and how each of the individuals involved experience and make sense of these interactions. It is proposed that in order to understand ‘resulting interactions’ more fully it is necessary to explore and compare the multiple perspectives of doctors, patients and others; including relatives and nurses. This thesis seeks to do this in an innovative way by reporting research, which involved observing and recording consultations between doctors and patients and their relatives and then conducting in-depth interviews with such people in order to explore their own insights into this process. In total, 115 episodes of data were collected and analysed from 16 patients and 16 doctors. The insights gained from this study are presented in relation to two main analytic themes; Doctors and Patients Acting their Parts, and Sharing Uncomfortable News. The data analysis highlighted a number of approaches used by patients and doctors to manage and control their interactions within the medical consultation. The implications of the study findings are discussed in relation to both wider theoretical perspectives and ideas for how doctors working in such settings could be assisted to consider alternative strategies for these aspects of their work.

The gastrointestinal (GI) hormone, gastrin, promotes cancer progression and its down-regulation has been linked to reduced cancer stem cell numbers. Gastrin acts through the cholecystokinin-2 receptor (CCK-2R) and its biological effects are blocked by CCK-2R inhibitors. We investigated the regulation of the CCK-2R and its potential role in promoting survival of cancer stem cells (CSC). A panel of cancer cell-lines, including GI, glioblastoma and lung, with CCK-2R-transfected cells as a positive control, were grown either as monolayers, or, to provide a 3D in vitro tumour model, as spheres. Linear-after-the-Exponential (LATE)-PCR was used to quantify CCK-2R gene expression and this was validated using siRNAs. Flow cytometry was used to investigate receptor protein expression. Activity of CCK-2R promoter reporters was quantified using luciferase assays. LATE-PCR for CCK-2R gene expression is 10,000-fold more sensitive than the Taqman-based assay, and provides a highly precise method for detection of genes which have important biological functions but low expression. This assay showed that primary non-small-cell lung tumours have significantly more expression than normal lung tissue, indicating a potential therapeutic marker. CCK-2R siRNAs resulted in up to 97% (p<0.05) knockdown of the receptor in cancer cells, confirming the specificity of LATE-PCR and offering a therapeutic possibility. The CCK-2R promoter constructs were active in lung, glioma and colorectal cancer cell-lines, demonstrating a potential drug target; however, transcriptional activity did not correlate with gene expression suggesting post-transcriptional or translational regulation is a factor affecting CCK-2R expression. Flow cytometry suggests the presence of a small population of cells within each of these cell-lines which expresses CCK-2R very highly, which was not correlated to CSC markers. However, CCK-2R expression was enriched when cells were grown as spheres, and inhibition caused a delay in sphere-forming, implying that the CCK-2R may play a role in tumour, and CSC, expansion. Thus, CCK2R provides a potential target for therapeutic intervention in cancer.

Alzheimer's disease (AD), the most common form of dementia, is a devastating age-related neurodegenerative disorder. There is a growing body of evidence suggesting that leading a stressful lifestyle is associated with a heightened risk of developing AD. This is supported by preclinical evidence using transgenic mice over-expressing genetic mutations leading to overt ß-amyloid protein production, a pathological marker of AD; stress in such mice has been capable of exacerbating AD-associated pathologies, including accelerating memory impairments and elevating ß-amyloid levels. In contrast, a recent study from our group demonstrated that a repeated mild stress procedure, novel cage stress, improved a short-term memory deficit and reduced the normal age-related increase in 3-amyloid levels. This thesis aimed to further characterise the beneficial effects of novel cage stress on AD-associated pathology in the TASTPM mouse model (double transgenic hAPP695swe x PS-I. M146V) which exhibits overt, age-related ß-amyloid pathology. First, age-related changes in AD- associated pathology, with or without exposure to novel cage stress, were assessed using a multidisciplinary approach incorporating measures of cognitive performance, in vivo magnetic resonance imaging and post-mortem analysis of ß-amyloid levels. The aim was to detect an age where we observe the most robust effect of stress; this time window was subsequently targeted to investigate a potential underlying mechanism, namely signalling though the glutamate alpha-amino-3-hydroxy-5- methyl-4-isoxazolepropionic receptor (AMPAr). Studies described throughout this thesis, alongside previously published data, indicate novel cage stress appears to improve AD-associated pathology in TASTPM mice, but independently of AMPAr function. It is likely that novel cage stress is insufficiently severe to induce detrimental effects, but, rather, subsequent repeated stimulation and physical activity may improve pathological status. A better understanding of lifestyle risk factors of AD, such as stress, will aid in identifying those at risk of developing the disorder. Moreover, discovering the underlying mechanisms linking stress with AD may open novel therapeutic avenues to treat the disorder.

This thesis presents a qualitative study of genetic counselling. Using a combination of semi-structured interviews and conversation analysis, it focuses on the role, function and structure of genetic counselling and on its status as medical or counselling work. Semi-structured interviews are used to ascertain genetic counsellors' accounts or perceptions of the nature of their role, their views on client expectations, and genetic counselling clients' perceptions and expectations of the same. Conversation analytic study of recorded genetic counselling consultations is used to identify whether or not they possess an overall shape and whether they appear conversationally as a counselling or a medical interaction. Rose's (1998, 1999) sociological work on the growth of the therapeutic community and the techne of 'psy' provides a framework for a discussion on the strength of the genetic counselling profession's association with a Rogerian counselling philosophy and on the potential difficulties this may bring. The questions are raised; does genetic counselling have many similarities to "personal, emotional or psychological" 'counselling' at all? And is this alliance with the counselling community either fair or possible for the professionals involved? The results were as follows. First, that the genetic counselling consultations in this corpus do not present with one unique overall shape that can encompass all interactions. Second, that the accounts of the genetic counsellors and clients in this sample, and the conversation analytic study of the recorded consultations, suggest that genetic counselling is primarily a medical-based activity and that this is what clients want. Third, that genetic counselling has a number of dissimilarities to psychotherapeutic counselling that suggest it is not so much 'counselling' as using counselling skills, and finally, that the tensions incurred in fulfilling medical-type tasks within what is ostensibly a 'counselling' role are neither fair nor practical for the professionals involved.

The alveolar epithelium is a key functional component of the air-blood barrier in the lung. Comprised of two morphologically distinct cell types, alveolar epithelial type I (ATI) and type II (ATII) cells, effective repair of the alveolar epithelial barrier following injury appears to be an important determinant of clinical outcome. The prevailing view suggests this repair is achieved by the proliferation of ATII cells and the transdifferentiation of ATII cells into ATI cells. Supplemental oxygen and mechanical ventilation are key therapeutic interventions in the supportive treatment of respiratory failure following lung injury, but the effects of hyperoxia and mechanical deformation in the injured lung, and on alveolar epithelial repair in particular, are largely unknown. The clinical impression however, is that poor outcome is associated with exposure of injured (repairing) epithelium to such iatrogenic ‘hits’. This thesis describes studies investigating the hypothesis that hyperoxia & mechanical deformation inhibit normal epithelial repair. The in vitro data presented demonstrate that hyperoxia reversibly inhibits the transdifferentiation of ATII-like cells into ATI-like cells with time in culture. Whilst confirming that hyperoxia is injurious to alveolar epithelial cells, these data further suggest the ATII cell population harbours a subpopulation of cells resistant to hyperoxia-induced injury. This subpopulation of cells appears to generate fewer reactive oxygen species and express lower levels of the zonula adherens protein E-cadherin. Using a panel of antibodies to ATI (RTI40) and ATII (MMC4 & RTII70) cell-selective proteins, the effect of hyperoxia on the phenotype of the alveolar epithelium in a rat model of resolving S. aureus-induced lung injury was investigated. These in vivo studies support the view that, under normoxic conditions, alveolar epithelial repair occurs through ATII cell proliferation & transdifferentiation of ATII cells into ATI cells, with transdifferentiation occurring via a novel intermediate (MMC4/RTI40-coexpressing) immunophenotype. However, in S. aureus-injured lungs exposed to hyperoxia, the resolution of ATII cell hyperplasia was impaired, with an increase in ATII cell-staining membrane and a reduction in intermediate cell-staining membrane compared to injured lungs exposed to normoxia alone. As hyperoxia is pro-apoptotic and known to inhibit ATII cell proliferation, these data support the hypothesis that hyperoxia impairs normal epithelial repair by inhibiting the transdifferentiation of ATII cells into ATI cells in vivo. The effect of mechanical deformation on alveolar epithelial cells in culture was investigated by examining changes in cell viability following exposure of epithelial cell monolayers to quantified levels of cyclic equibiaxial mechanical strain. In the central region of monolayers, deformation-induced injury was a non-linear function of deformation magnitude, with significant injury occurring only following exposure to strains greater than those associated with inflation of the intact lung to total lung capacity. However, these studies demonstrate for the first time that different epithelial cell phenotypes within the same culture system have different sensitivities to deformation-induced injury, with spreading RTI40-expressing cells in the peripheral region of epithelial cell monolayers and in the region of ‘repairing’ wounds being injured even at physiological levels of mechanical strain. These findings are consistent with the hypothesis that alveolar epithelial cells in regions of epithelial repair are highly susceptible to deformation-induced injury.

No description available.

No description available.

<p> Tan spot and Septoria nodorum blotch cause serious yield losses in winter wheat in North Dakota as the majority of commercially grown cultivars are susceptible. This study aimed to identify lines with improved resistance for use as breeding parents. First, advanced NDSU breeding lines and alternative sources of resistance were inoculated with fungal isolates and tested for necrotrophic effector sensitivity. Second, resistant lines were derived from a highly heterogeneous recurrent mass selection F₂ population using single seed descent inbreeding coupled with selection for resistance. Finally, the best performers from both experiments (total of 52 lines) were evaluated to confirm resistance. In addition the 52 lines were analyzed with markers that detect <i>Tsn1</i> and the 1RS rye translocation. Twenty lines were identified with simultaneous resistance to four or three fungal isolates and insensitivity to three, two, or one necrotrophic effectors (of which 11 can be used directly as new parents).</p>

<p>Osteoarthritis is one of the most common, debilitating, musculoskeletal diseases in the world. Currently, there is no cure. It is well-known that a traumatic, joint injury increases the risk of developing post-traumatic osteoarthritis (PTOA). Therefore, in order to improve clinical treatment and prevention strategies for post-traumatic osteoarthritis (PTOA), a series of translational studies were conducted to develop research models to evaluate the effects of impact injury. </p><p> The first section of this dissertation (Ch. 1&ndash;2) provides a comprehensive introduction and literature review related to both clinical PTOA as well as previous research investigations of PTOA. The second section of this dissertation (Ch. 3&ndash;6) describes the methodology of optimizing a servo-hydraulic test machine to deliver a controlled impact injury (Ch. 3) as well as subsequent studies using this device to injure articular cartilage (Ch. 4) and cartilage-bone explants (Ch. 5&ndash;6). Further, the effects of dynamic, compressive loading to mimic walking after impact injury of cartilage-bone explants was investigated (Ch. 6). The third section of this dissertation (Ch. 7&ndash;8) details the development of an impactor device that may be used for pre-clinical, animal models. </p><p> Many significant findings were discovered through this dissertation work. Specifically, by using the proportional-integral-derivative (40, 0, 0) values, a large (25kN) servo-hydraulic test machine may be used to deliver a controlled impact injury to explants (Ch. 3). Biomarkers glycosaminoglycan (GAG) and prostaglandin E2 (PGE₂) were elevated after cartilage impact injury with PGE2 having the highest mechanosensitivity than any other biomarker (Ch. 4). Energy absorbed during cartilage-bone injury is dependent upon trauma severity; PGE₂ and monocyte attractant protein (MCP-1) were elevated following cartilage-bone injury (Ch. 5). Dynamic, compressive loading retained cell viability in non-impacted cartilage-bone explants and mitigated GAG release in impacted explants; GAG and PGE₂ were elevated due to cartilage-bone injury whereas matrix metalloproteinase-2 (MMP-2) and interleukin-8 (IL-8) were elevated due to injury plus dynamic, compressive loading (Ch. 6). The development of a 8mm diameter impactor does create articular cartilage damage (Ch. 7), albeit a smaller, 2mm diameter impactor creates higher impact stresses and may be used arthroscopically for pre-clinical animal models (Ch. 8). </p>