Poster presentations of Congressus Pharmaceuticus Hungaricus XVI.

Abstracts of poster presentations of Congressus Pharmaceuticus Hungaricus XVI

Usability analysis of contending electronic health record systems

In this paper, we report measured usability of two leading EHR systems during procurement. A total of 18 users participated in paired-usability testing of three scenarios: ordering and managing medications by an outpatient physician, medicine administration by an inpatient nurse and scheduling of appointments by nursing staff. Data for audio, screen capture, satisfaction rating, task success and errors made was collected during testing. We found a clear difference between the systems for percentage of successfully completed tasks, two different satisfaction measures and perceived learnability when looking at the results over all scenarios. We conclude that usability should be evaluated during procurement and the difference in usability between systems could be revealed even with fewer measures than were used in our study. © 2019 American Psychological Association Inc. All rights reserved.Peer reviewe

The optometric control of myopia

A doctoral dissertation reviewing the current literature concerning the classification, etiology, and treatment of myopia. Five etiological theories are discussed, they include; genetic theory, normal biological variation theory, conditions of use theory, holistic theory, and deprivational theory. Treatment options were classified as being structural or functional in theory. Structural treatment options reviewed include; concave lens prescription, medical treatment, orthokeratology, and refractive surgery. Functional treatment options reviewed include; nearpoint lens prescription, vision therapy, holistic therapy, and biofeedback & visual acuity training. An integrative I holistic treatment model drawing from various treatment theories was developed as a means to better control myopia through the use of all treatment options available to the optometrist. The dissertation concludes that the optometric profession should look beyond the one dimensional treatment strategy of prescribing concave compensatory lenses when treating myopia. More emphasis needs to be given to preventative vision care, and control of myopic progression. Further clinical research investigating the efficacy of such a treatment plan is needed

Integrative physiology of human aerobic fitness and the influence of the ACE I/Dp genotype

The search for genes that influence human performance and health constitutes a popular topic of current research. One such genetic constituent that has caused much interest over the last 20 years is the angiotensin converting enzyme insertion/deletion polymorphism (ACE I/Dp). There is much controversy in the literature regarding the role (if any) of this polymorphism as effects and effect size vary between populations of different origin and training status. The aim of this thesis was to analyse at the whole organism level whether skeletal muscle plasticity explains the association of the ACE I/D polymorphism with metabolic fitness. Regular endurance exercise reduces the risk of a plethora of diseases, but the exact molecular mechanisms are not fully understood – the ability of muscle to adapt to exercise stimulus is key. Trained individuals demonstrated clear physiological differences of aerobic processes such as increased oxygen usage, greater power output and reduced body fat that would be expected (T-Test: p<0.001). By contrast, when examining metabolite changes, at rest, in the local muscle there were few (n=6) nonpolar (lipid species) metabolite (assessed by mass spectrometry) differences between the trained and untrained. However, after an acute exercise bout working muscle in trained individuals displayed a significant up-regulation of (n=76) non-polar metabolites (Repeated ANOVA: p=0.0004), illustrating that training produces significant adaptations in substrate metabolism at the local level. Would there be a genetic component contributing towards these physiological and local muscle differences? Individuals with the ACE I-allele (insertion sequence) had increased capillary density, and there were significant differences in transcripts, together with both polar and non-polar metabolites in the untrained population at rest and following an acute exercise bout. These differences were lost in the trained population. In a different population (Swiss) capillary density was increased following a training programme in the absence of the I-allele – in contrast to the other population (British). However, gene expression response of important factors, to exercise was preserved. In conclusion, a trained population demonstrated enhanced non-polar metabolism in the working muscle after an exercise bout, and the dominant stimulus of regular exercise over-rides the influence of the ACE I/Dp. Nurture over-rides nature

Mass spectrometry metabonomics of HIV-1 sera

Background: Metabolic complications resulting from the human immunodeficiency virus type 1 (HIV-1) and the acquired immunodeficiency syndrome (AIDS) are as common as the immune system disruption caused by the virus, but not as well known. Highly active antiretroviral therapy (HAART) used to treat HIV-1 infection exacerbates the effects HIV-1 has on the host‘s metabolism. Common metabolic complications such as insulin resistance, lipodystrophy, lactic acidosis and others contribute to morbidity and mortality during HIV/AIDS. The detection of HIV-1 related metabolic biomarkers assists in diagnosing and monitoring metabolic complications, however, limitations of the conventional methodologies used for detecting these molecules caused a paucity of data on HIV-related metabolic indicators. Metabonomics, the ability to measure multiple metabolites simultaneasously, shows promise in distinguishing HIV-1 negative and positive patients through nuclear magnetic resonance (NMR) and vibrational spectroscopy as well as mass spectrometry (MS) profiles of various biofluids. The objective of this study was to determine the abilities of ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) in the identification of serum metabolites associated with HIV and/or HAART. The study was also aimed at determining whether data from a less sensitive technique, Fourier transform infrared (FTIR) spectroscopy would lead to comparable conclusions as those derived using UPLC-MS data. Methods: Sera were collected from three experimental groups; HIV negative (n=32), HIV positive (n=29) and HIV positive patients receiving HAART (n=34). Metabolites were extracted using a conventional approach of cold methanol extraction as well as the OstroTM plate extraction technology which involved filtration by positive pressure. The filtrate was analysed in the negative and positive electrospray ionization (ESI) modes of UPLC-MS. Serum samples were also dried overnight and analysed using FTIR. Data processing and chemometric analysis was carried out using the SPSS 19.0 and MassLynx v4.1 software packages. Following extensive statistical evaluation of data, bioinformatics approaches that assisted with metabolite identification were conducted. Results: The combination of OstroTM plates and UPLC-MS produced high resolution chromatograms that showed visible differences among the serum samples of HIV negative, HIV positive and HIV positive patients receiving HAART. Linear discriminant analysis (LDA) classified experimental groups into the correct categories with great accuracy (>88%), using potential biomarkers responsible for the observed group variations. Principal component analysis (PCA) showed clear separations as well as some overlap among the three experimental groups. Orthogonal projections to latent structures-discriminant analysis (OPLS-DA) showed clear differences between two classes of samples at a time and potential biomarkers were selected from accompanying S-Plots. Hundred and twelve distinct group distinguishing metabolites detected from both ESI positive and ESI negative modes were significantly altered (p<0.05). HIV and/or HAART altered metabolites of energy, neuronal and mitochondrial processes were identified and were evident in the amino acid, carbohydrate, lipid and nucleoside/nucleotide metabolic products being detected. Antiviral drugs [mostly nucleoside/nucleotide reverse transcriptase inhibitors (NRTIs)], antiinflammatory and anti-cancer drugs, nutrient supplements and other drugs associated with the treatment of HIV conditions also contributed to class variations. FTIR generated metabolic patterns that separated the three experimental groups on an LDA scatter plot which achieved correct (>87%) classification accuracy. The significantly altered patterns indicated hydroxyl and alkene group vibrations and these groups were present in the metabolites identified by MS. Conclusion: OstroTM plates and UPLC-MS successfully purified, detected and identified sera metabolites distinguishing HIV and/or HAART patients. The different statistical analysis methods applied in this study were in agreement and the OPLS-DA statistical tool complemented the sensitivity of UPLC-MS for the detected distinguishing metabolites. The approach employed here delivered promising findings for use in the discovery of metabolic biomarkers. Distinguishing metabolites identified could be traced to HIV-infection and/or treatment. Findings from this study corroborated with others which showed that NRTIs remain a challenge in the era of HAART toxicities, especially their dominant effect on mitochondrial dysfunction. This work therefore suggests the use of UPLC-MS in HIV disease diagnosis, prognosis, monitoring of treatment success or failure and the ability to link treatment to metabolic complications. Even though FTIR is less sensitive than UPLC-MS, it was successful in detecting metabolic patterns that corresponded to some metabolites detected by UPLC-MS. This suggests that this easier to perform technique also has potential clinical application in monitoring HIV/AIDS.Dissertation (MSc)--University of Pretoria, 2015.South African National Research Foundation (NRF)Medical Research Council (MRC)Technology Innovation Agency (TIA)BiochemistryMScUnrestricte

Phenotypic heterogeneity in high-grade serous ovarian cancer: evidence for a stochastic model of proliferation

High-grade serous ovarian cancer (HGSOC) represents the largest and most lethal subtype of ovarian cancers. Chemotherapy is effective at depleting these tumours, sometimes eliminating all visible disease, however the tumour invariably regrows from surviving tumour cells. Improving treatment depends on understanding and targeting these residual cells. The bulk of HGSOC tumour cells strongly express the epithelial surface marker EpCAM. However, previous work in the Brenton and Stingl Laboratories identified a distinct subpopulation of cells with low levels of EpCAM. These cells are uniquely able to regrow tumours in the lab, and are enriched by chemotherapy. The objective of this project was to understand the dynamics of these cells, not during transplantation or initiation, but under conditions similar to an established tumour in the patient. To this end, tumours were engrafted into immune-deficient mice and allowed to grow. These were then studied only once they had become established. Nucleoside tracing studies were used to assay proliferation within each of the subpopulations. These were performed both over the short-term (to determine subpopulation proliferation rate), and over the long-term (to determine resulting expansion of the subpopulations over time). The results show that the subpopulation deficient in EpCAM is seen to divide more slowly in all cases. Both subpopulations divide stochastically, and their rates of proliferation match the expansion of each subpopulation. This indicates that both subpopulations are self-supporting. Inducible lineage tracing tools were validated to investigate hierarchy and subpopulation interconversion. Unfortunately, the tools generated for this purpose did not prove reliable and solutions were not available within the scope of this project. An alternative approach was also tested, assessing the symmetry of cell division by immunofluorescence, but is not yet optimised. To provide a deeper understanding of these phenotypes, single-cell-mRNAseq was performed. Sequencing is only half complete, but initial analysis already appear to indicate distinct gene expression patterns between cell clusters corresponding to observed phenotypic subpopulations. This provides a basis for future work into targeting the resistant subpopulation. In conclusion, this work refutes the proposed cancer stem cell model in HGSOC and presents evidence of stochastic division and self-supporting dynamics in distinct phenotypic subpopulations. It provides evidence of a distinct subpopulation, low in EpCAM, which is slow-cycling. This further supports the idea that these cells are the origin of relapse. In addition, this work begins the process of assessing (at single-cell level) differential expression in this fraction, elucidating the molecular nature of this subpopulation, and guiding future work to target these cells.Funded by Cancer Research U

The influence of helminths on immune responses to HIV.

Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2009.In South Africa, co-infection with HIV and intestinal parasites is a major challenge in disadvantaged communities who live in densely populated under-serviced urban informal settlements. This pilot cross sectional study evaluates the immunological effects of co-infection with Ascaris lumbricoides and Trichuris trichura on the immune response to HIV. The work was a substudy of a prospective double blind, placebo-controlled investigation to test whether regular deworming changes the immune profile of HIV positive individuals with concurrent helminth infection. The substudy has a cross sectional design and presents pilot data that defines immune profiles of HIV-1 positive individuals with and without gastrointestinal helminth (Ascaris lumbricoides and Trichuris trichura) infection. The hypothesis was that concurrent helminth infection adversely affects immune responses against HIV. It was conducted in an area of high helminth endemnicity and limited infrastructural resources. Individuals with known HIV infection were recruited from an HIV Support Group and HIV negative individuals residing in the same area (for demographic matching) were used for comparison. The substudy was to provide pilot data for future larger scale and possible interventional studies. The current work is limited by the cross sectional design, moderate sample size and practical challenges. The profile of lymphocyte phenotypes, viral loads, eosinophils, activation markers, expression of the nuclear proliferation antigen-Ki67 and activation regulator antigen CTLA-4 were analysed using flow cytometry in HIV positive and negative subgroups with or without helminth infection. The type-1, type-2 and inflammatory cytokines were analysed using multiplex cytokine array technology. These were correlated with immune responses to HIV. Non parametric statistics were used to describe differences in the variables between the subgroups. A major finding of the study was the result of the supplementary use of the serological marker, Ascaris lumbricoides-specific IgE in addition to the presence (or absence) of helminth eggs in stools to classify intestinal helminth infection status. Two significant outcomes of this measure were the enhancement of diagnosis of current or recent helminth infection and, more importantly, the distinction of different phenotypes of individuals who displayed different immunological responses to co-infection with HIV and helminths. The different helminth infection phenotypes are defined by stool egg positivity (egg⁺) or negativity (egg⁻) with either high or low Ascaris-specific IgE (IgEhi or IgElo) respectively. The four subgroups, egg⁺IgEhi, egg⁺IgElo, egg⁻IgEhi and egg⁻IgElo showed different interactions with regards to immune response to HIV. It should be noted that no Trichuris specific IgE tests are commercially available but that there is significant antigenic cross-reactivity with Ascaris antigen. The presence of helminth stool eggs and high Ascaris IgE (egg⁺IgEhi) was associated with the following characteristics: reduction in numbers of all lymphocyte populations, frequent eosinophilia, highly activated immune profiles, antigen specific proliferative hyporesponsiveness, impaired type 1 cytokine responses in unstimulated and antigen stimulated cells and increased TNFα levels. In HIV infected individuals, the egg⁺IgEhi helminth infection status was associated with lower but not significant CD4⁺ counts and higher viral loads. A strong negative correlation was observed between viral loads, CD4⁺ and CD8⁺ cells in this subgroup. Subgroups with high IgE (egg⁺IgEhi and egg⁻IgEhi) had elevated Th2 markers with lower CD4⁺ counts and higher viral loads in the HIV⁺ group. The inverse correlation between viral load and CD4⁺ counts found in all the HIV⁺ participants was strongest in these two subgroups. Individuals with parasite eggs in stool and low Ascaris IgE (egg⁺/IgElo) presented a modified Th2 profile. This subgroup had high absolute numbers of all lymphocyte subsets in both HIV⁻ and HIV⁺ groups with higher CD4⁺ counts in the HIV⁻ and lower viral load in the HIV⁺ groups as well as higher interferon gamma, lower IL-4 and higher IL-10. In conclusion, the results suggest that helminth infections may be associated with deleterious effects on the immune responses to HIV in certain groups of susceptible individuals. The underlying reasons for the different stool egg/Ascaris IgE combinations in settings with high exposure to helminthes is currently not clear but genetic predisposition and environmental factors could play a role. Future studies of helminth- HIV co-infection have to ensure adequate definition of helminth infection status by the use of both stool examination and measurement of helminth-specific IgE as the infection phenotype is associated with differential effects on HIV associated immune responses. This may also apply to co-infection with other pathogens, including tuberculosis. The long-term effect of helminth co-infection in HIV positive people was not assessed in this study but requires further studies

British Military Veterans and the Criminal Justice System in the United Kingdom: Situating the Self in Veteran Research

The 21st Century has seen the continuation of armed conflict, exposing military personnel to the rigours of warfare and the challenges of transition back to a civilian identity. There has been a renewed realisation that there exists a sub-group within the criminal justice system (CJS) of veterans and whilst the exact figures are debated, their presence is not. This thesis seeks to capture the perspectives and experiences of veterans who are identified as exoffenders and those having been employed in the CJS as practitioners. The super-structuralist concept of the CJS collectively represent services of a ‘total institution’ that have shared similarities and differences to life within the ‘total institution’ of the Armed Forces. The life stories of the participants indicated that whilst one veteran life story trajectory (veteran practitioner) appeared to be able to adapt during the transition to a civilian identity successfully, there was evidence that the other veteran life story trajectory (veteran exoffender) found themselves segregated and isolated from a familiar veteran identity with few resources to survive the experience unscathed. This exploratory qualitative study provides emancipatory evidence that the process of entering the CJS as offenders often fails to address the origins of their criminal behaviour or from the wider social context that creates a cyclical response. The veteran practitioners appear to hold a crucial insight into the issues and seek to progress the CJS’s need to expand its knowledge base on the identification, diversion and management of veteran offenders. The study was theoretically informed through the use of reflexivity to articulate the internal and external dialogue of what is known and how it is known in understanding the lived experiences of 17 participants. Life stories were collected from in-depth interviews across the United Kingdom. The life stories were analysed thematically, providing insight and understanding through the elicitation of narratives derived from the contours of meaning from the participants’ (veterans) experiences and enunciating the two separate life story trajectories into the CJS. The findings of this study indicate the participants need to belong and explores how their veteran identity instilled in them both a source of strength and a feeling of anguish, as their new lives could not offer the same security and sense of belonging. The negative consequences of being identified as an offender often resulted in the emergence of stigma and associated shame upon themselves and their families. The life stories demonstrated disparities between the attempted empowering philosophies of the veteran practitioners and the practices imposed generally by the CJS. There were numerous examples of how the veterans’ prior exposure to the institution of the Armed Forces had shaped their experiences and engagement with the institutions of the CJS. Both sub-groups of veterans constructed positive ownership of their veteran identity which at times served to counterbalance their negative experiences of transition from military to a civilian identity. These constructions of their experiences highlight the vulnerability of this sub-group within the CJS and the failure of the system and wider society to address the consequences of military service on some veterans. This research raises the issue of the ‘fallout’ from the recruitment of youth from communities where established socio-economic deprivation has created fertile recruitment grounds for the Armed Forces. The analysis identifies a pragmatic need to address the gaps within the research literature as well as multi-agency working, in order to expand veteran peer support schemes. The voice of the veteran has been overlooked within the positivist research approach, this study seeks to capture the viewpoint of the veterans through reflexive exploratory research undertaken by a veteran researcher to understand the phenomena. Researching the experiences of veterans’ experiences of the CJS presented ethical and methodological challenges. The study has provided new knowledge and understanding that can be disseminated and used to improve current practices and policies