The vascular response in chronic periodontitis

This thesis describes work done at the Institute of Dental Research in Sydney between February of 1986 and January 1990. The broad subject of the work is the role of vascular endothelial cells (ECs) in chronic inflammation. Periodontitis has been used as an example of chronic inflammatory disease, and provides the focus for this study of endothelial biology. In Chapter 1, aspects of the endothelial literature which provide relevant background information for work described in later chapters are reviewed. In Chapter 2, literature relating to aetiology and pathogenesis of chronic inflammatory periodontal disease is discussed. To maintain relevance of literature reviews to experimental work, each subsequent chapter contains a small literature review of material relating to the subject of the specific chapter. Early laboratory work is described in Chapter 3, and consisted of a morphological survey of the vascular changes occurring in gingival tissues with development of chronic periodontitis. Expansion of the vasculature and appearance of phenotypically specialised high endothelial cells (HECs), were associated with progression of the disease. Vessels with HECs and had a similar appearance to those known to be responsible for lymphocyte recirculation described in lymphoid tissues and chronic inflammatory sites. In the course of performing this survey, a perivascular hyaline material was noted surrounding capillaries close to the bacterial plaque irritant. The incidence, distribution, extent, ultrastructre and immuo-histochemistry of this material was more closely investigated, and the possible pathogenesis and significance of the material discussed in Chapter 4. In Chapter 5, the ultrastructural, histochemical and functional properties of gingival HECs are described, and compared with the well characterised HECs of rat lymph nodes. It was found that periodontal vessels were very similar to those in rat lymph nodes, with the exception however, that the gingival vessels appeared to exchange polymorphonuclear leukocytes almost exclusively, while vessels with HECs in lymph nodes and other locations are known as sites of lymphocyte recirculation. This observation indicated that the function of HECs requires further investigation, with particular regard to the synthetic activity of the cells. HECs were consistently alkaline phosphatise (AP) negative. The negative association between leukocyte emigration and AP activity (APA), as well as evidence in the literature illustrating both the wide substrate specificity of this enzyme and the importance of phosphorylation in the control of protein activation, suggested that AP could play a role in regulating leukocyte emigration. A pre-requisite for the investigation of this possibility, is the identification of a rich source of the identical iso-enzyme of AP to what is present in ECs. In Chapter 6, the sensitivity of endothelial AP to a panel of inhibitors is compared with that of a number of tissues for which isoenzyme has been identified. Endothelial AP was identified as the liver/bone/kidney isoenzyme. This allows the use of kidney tissue as a relevant source of AP for use in further study of the role of this enzyme in EC biology. It was clear that in order to study both the synthetic activity of HECs, as well as the role of AP in the control of leukocyte emigration, a method for obtaining high density primary cultures of HECs had to be established. Chapter 7 describes work done towards the development of such a culture system. The availability in the latter phase of the work of suitable probe for the technique of the in-situ hybridization allowed the possibility of testing the hypothesis that HECs are important cytokine producers. It was felt that this would provide some basis for the further study of those cells in-virtro. This work is described in the appendix. The general discussion in Chapter 8, summarises the work, and develops potential areas of study arising from the finding of this thesis

The vascular response in chronic periodontitis

Dissertação de Mestrado Integrado em Arquitectura, apresentada ao Departamento de Arquitectura da Faculdade de Ciências e Tecnologia da Universidade de Coimbra, sob a orientação do Prof. Doutor Walter Rossa.Integrada num trabalho mais abrangente que simula uma candidatura de Coimbra a Capital Europeia da Cultura 2027, a dissertação esboça um plano de regeneração urbana do Pólo I da Universidade de Coimbra, detentor de uma vertente simbólica e funcional de produção cultural/artística actualmente subaproveitada, porque ignorada. A construção de mais equipamentos é difícil - dada a configuração física do existente e as condicionantes do estatuto UNESCO - mas sobretudo desnecessária, dadas as valências espaciais e funcionais do conjunto, a nível de espaços interiores e exteriores. Trata-se da requalificação geral e dinamização do espaço público, que inclui a intervenção no Largo D. Dinis, Largo da Feira e Pátio das Escolas, lugares-chave de articulação dos espaços e ritmos da Alta universitária. Há uma contradição entre esta variedade espacial e a monofuncionalidade que caracteriza o desenvolvimento urbano da Alta no último século (sobretudo no Pólo I), que reflecte a tendência de dispersão da instituição pela cidade, e a recusa da densificação. Esta questão está intimamente associada à do património, um tema que aqui é basilar, embora implícito, e a posição que assume é a do esforço e estímulo pelo património activo e produtivo, em oposição àquele inerte, lucrativo e parado no tempo. As ideias de plano e processo tornam-se então essenciais e orientam as reflexões e decisões do trabalho, que prioriza o planeamento a longo prazo, capaz de satisfazer tanto os momentos temporários como o quotidiano permanente (duplamente proveitoso), fundamental para se assegurar o desenvolvimento de uma cidade com autonomia própria, não condicionado pelos momentos e modas que este tipo de iniciativas são. Análise e proposição confundem-se - no processo de trabalho e no resultado final - mas é possível ver na estrutura geral dois momentos. Um primeiro de leitura e interpretação de documentos pertinentes, de forma a compreender o historial da iniciativa em termos dos seus objectivos, métodos e abordagens a questões semelhantes às que aqui se tratam, e definir um breve retrato cultural coimbrão - que gera este contexto virtual, enquadra o trabalho e o torna relevante. Um segundo de análise espacial e funcional do Pólo I, das estruturas e infraestruturas que com ele se relacionam directa e indirectamente, para determinar uma estimativa de características, capacidades e adequabilidade do lugar e sua envolvente imediatamente próxima, expondo-se ao longo do trabalho propostas nas mais diversas áreas e níveis de intervenção.Part of a broader work that simulates a candidacy of Coimbra to European Capital of Culture 2027, this essay seeks to draft an urban renewal plan for the main University of Coimbra’s hub, which holds a symbolic and functional side of cultural/artistic production currently underused, because ignored. The construction of new buildings is hardly possible - because of the site’s physical situation and the constraints brought by the UNESCO status - but above all unnecessary, given the spatial and functional characteristics of the ensemble, both in the interior and exterior domains. Overall it’s about requalifying and regenerating public space, which includes interventions in Largo D. Dinis, Largo da Feira and Pátio das Escolas, key places in articulating the spaces and rhythms of the university’s oldest and central hub. There is a contradiction between this spatial variety and the monofunctionality that characterizes the urban development of this zone in the last century (especially the campus), which reflects a tendency for the institution’s sprawl over the city, and the denial of densification. This issue is closely related to the discussion of heritage, a fundamental theme here, even though implicit, and the position it takes is one of effort and stimulus for an active and productive heritage, as opposed to one that is numb, profitable and frozen in time. The concepts of plan and process become thus essential, guiding the work’s thinking and its decisions, that prioritize long-term planning, able to satisfy both temporary moments and the permanent everyday (twice as fruitful), essential to ensure a city’s development which is autonomous, and thus not influenced by moments and trends that this kind of initiatives really are. Analysis and proposal intermingle - both in the work process and the final result - but it is possible to tell apart two moments in the overall structure. A first one of reading and interpretation of relevant documents, as to understand the history of this particular iniciative in terms of its objectives, methods and approach to issues similar to the ones dealt with here, and tracing of a cultural portrait of the city - that generates this virtual context, which frames the work and makes it relevant. A second one of spatial and functional analysis of the university’s main hub, of the structures and infrastructures that directly and indirectly relate themselves with it, in order to estimate the characteristics, capacities and suitability of the place and its surrounding environment, while proposals are constantly presented throughout, concerning diverse themes and intervention levels

Supplementary Data for the Paper: Description of comprehensive dental services supported by the Medicare Chronic Disease Dental Scheme In the first twenty three months of operation, In Press: The ANZ Journal of Public Health

The two tables provided show details at the state and territory level, of information presented only at the National level in Tables 1 and 2 of the paper entitled: Description of comprehensive dental services supported by the Medicare Chronic Disease Dental Scheme In the first twenty three months of operation, which is published by the Australian and New Zealand Journal of Public Health (2011, In Press). Tables show: The number of Medicare Chronic Disease Dental Scheme supported patients and services according to treatment type (Table 2); and The average number of Medicare Chronic Disease Dental Scheme services per patient according to type of service delivered, expressed as both services per patient and the relative percentage of services per patient (Table 3). Details on how this data was collected, as well as how it was interpreted and other relevant information are available in the complete manuscript published by the ANZJPH, 2011 (In Press)

Description of comprehensive dental services supported by the Medicare Chronic Disease Dental Scheme in the first 23 months of operation

Abstract Objective: Australia's Medicare universal insurance system has supported comprehensive dental service through the Chronic Disease Dental Scheme (CDDS) since November 2007. Public debate opposing CDDS includes claims of over‐servicing, calls for expansion to universal eligibility, and government threat of closure. Here we examine CDDS services over the first 23 months of operation. Methods: CDDS statistics on patient age, gender and item numbers claimed from November 2007 to December 2009 from Medicare were subjected to analysis. Results: The distribution of 404,768 total CDDS patients varied across Australia from 3.6% of the population in NSW to 0.07% in NT, while uptake increased over time. The average patient had 7.58 dental treatments, and the most common were: direct restorations (2.27), preventive and periodontal services (1.46), diagnostic services (1.43), extractions (0.77), and new dentures (0.53). Crown and bridgework appeared over‐represented (0.48). Conclusion: Although data do suggest over‐servicing in crown and bridgework, there also appears to be significant community need for the CDDS. Implication: Clear guidelines for dental clinical diagnosis and treatment planning, as well as a pre‐approval process for crown and bridgework is suggested to improve the CDDS, and this could form the basis for expansion to universal eligibility for dental Medicare

Data files of work described in the manuscript: Increased cell size, structural complexity and migration of cancer cells acquiring fibroblast organelles by cell-projection pumping

These are data files for work described in the manuscript: Increased cell size, structural complexity and migration of cancer cells acquiring fibroblast organelles by cell-projection pumping. At the time these files were uploaded to the University of Sydney Research Repository, this work was in-press in the open-access on-line journal PLoS One. A pre-print of the manuscript was also available at: bioRxiv doi: http://biorxiv.org/cgi/content/short/770693v1. FACS, proliferation and migration data are provided in two separate MS Excel files titled: 1) Migration Data: which contains calculations for distance migrated in scratch assays; 2) FACS Proliferation and Scatterplots: which contains data for FACS forward and side scatter analysis, as well as proliferation assays and scatterplots for all data

Increased cell size, structural complexity and migration of cancer cells acquiring fibroblast organelles by cell-projection pumping.

We recently described a hydrodynamic mechanism for cytoplasmic transfer between cells, termed cell-projection pumping (CPP). Earlier image analysis related altered SAOS-2 osteosarcoma cell morphology, to what we now recognize as CPP uptake of fibroblast cytoplasm. We here examine SAOS-2 phenotype following co-culture with human dermal fibroblasts (HDF) in which organelles were pre-labelled with a fluorescent lipophilic marker. Fluorescence activated cell sorting (FACS) analysis was performed of HDF and SAOS-2, cultured either alone or together. FACS forward scatter is proportionate to cell size, and increased for SAOS-2 with high levels of HDF fluorescence uptake (p < 0.004). FACS side scatter is proportionate to internal cell complexity, and increased in SAOS-2 with increasing uptake of HDF fluorescence (p < 0.004), consistent with uptake of HDF organelles. Scratch migration assays revealed that HDF migrated more quickly than SAOS-2 in both isolated cell culture, and following co-culture (p < 0.004). Notably, SAOS-2 with high levels of HDF labelling migrated faster compared with SAOS-2 with low HDF labelling (p < 0.008). A slight and unconvincing reduction in SAOS-2 proliferation was seen (p < 0.02). Similar results were obtained in single additional experiments with A673 and H312 cancer cells. Forward and side scatter results suggest organellar transfer by CPP increases cancer cell morphological diversity. This may contribute to histological pleomorphism relevant to cancer diagnosis and prognosis. Also, increased migration of sub-populations of cancer cells with high CPP organellar uptake, may contribute to invasion and metastasis in-vivo. We thus suggest relevance of CPP to cancer diagnosis and progression

Economic benefit of a polyacrylate-based hydrogel compared to an amorphous hydrogel in wound bed preparation of venous leg ulcers

Objective: To assess the cost-effectiveness of a polyacrylate (PA)-based hydrogel compared to an amorphous hydrogel in wound bed preparation for venous leg ulcers. Method: A cost-effectiveness analysis was undertaken alongside a multicenter, randomized controlled trial performed in France. A total of 75 patients with venous leg ulcers extensively covered with fibrin and necrotic tissue were randomized to a PA-containing hydrogel or an amorphous hydrogel. Wounds were treated for 14 days and costs were estimated from the - German payer's perspective. Medical costs included study treatment, wound treatment - supply, and labor time. The clinical benefit was expressed as the number of patients with wounds.50% covered with granulation tissue within 14 days. The incremental cost-effectiveness ratio (ICER) was expressed as the additional cost spent with >50% granulation tissue per day per patient within 14 days of leg ulcer care. Results: Because of individual pricing of wound dressings in hospitals, cost data were derived from the outpatient sector. A total of 33 patients were treated using the PA-based hydrogel and 37 patients using the amorphous hydrogel. The estimated total direct costs per patient and per 14 days of therapy were (SIC)306 for both treatment groups. However, with the PA-based hydrogel, 2.5 additional days with wounds covered.50% with granulation tissues were gained within 14 days of leg ulcer care compared to the comparator. The ICER was (SIC)0 per additional day spent with >50% granulation tissue. Conclusion: Although there were a greater number of dressing changes in the PA-based hydrogel treatment, the total treatment cost for 14 days of leg ulcer care was the same for both the PA-based-hydrogel and amorphous-hydrogel treatment modalities. The cost benefit favored the PA-based hydrogel because of its greater clinical efficiency in producing a viable wound bed