This study looks at human movement in order to formulate a principle that can potentially enrich the understanding of interior design and spatial awareness within it. It considers the spatial identity of the dis/abled body by looking at the importance of the embodied experience in relation to environment. The body has dynamic abilities; therefore it is an instrument for creating form. This tangible form in context with interior design can inform our understanding of spatial needs and be used as a design-informing tool. Specifically this project uses spatial identity as a theory to guide the conceptual ideas and as a practical tool to design a work environment and showroom for a client with Thrombocytopenia Absent Radius (TAR) syndrome. / October 2007

This study looks at human movement in order to formulate a principle that can potentially enrich the understanding of interior design and spatial awareness within it. It considers the spatial identity of the dis/abled body by looking at the importance of the embodied experience in relation to environment. The body has dynamic abilities; therefore it is an instrument for creating form. This tangible form in context with interior design can inform our understanding of spatial needs and be used as a design-informing tool. Specifically this project uses spatial identity as a theory to guide the conceptual ideas and as a practical tool to design a work environment and showroom for a client with Thrombocytopenia Absent Radius (TAR) syndrome.

This study looks at human movement in order to formulate a principle that can potentially enrich the understanding of interior design and spatial awareness within it. It considers the spatial identity of the dis/abled body by looking at the importance of the embodied experience in relation to environment. The body has dynamic abilities; therefore it is an instrument for creating form. This tangible form in context with interior design can inform our understanding of spatial needs and be used as a design-informing tool. Specifically this project uses spatial identity as a theory to guide the conceptual ideas and as a practical tool to design a work environment and showroom for a client with Thrombocytopenia Absent Radius (TAR) syndrome.

Immune heparin-induced thrombocytopenia (HIT) is a potentially serious complication of heparin therapy and is associated with antibodies directed against a complex of platelet factor 4 (PF4) and heparin. Early diagnosis of HIT is important to reduce morbidity and mortality. I developed an enzyme immunoassay that detects the binding of HIT IgG to PF4-heparin in the fluid phase. This required techniques to purify and biotinylate PF4. The fluid phase assay produces consistently low background and can detect low levels of anti-PF4-heparin. It is suited to testing alternative anticoagulants because, unlike in an ELISA, a clearly defined amount of antigen is available for antibody binding. I was able to detect anti-PF4-heparin IgG in 93% of HIT patients. I also investigated cross-reactivity of anti-PF4-heparin antibodies with PF4 complexed to alternative heparin-like anticoagulants. Low molecular weight heparins cross-reacted with 88% of the sera from HIT patients while half of the HIT sera weakly cross-reacted with PF4-danaparoid (Orgaran). The thrombocytopenia and thrombosis of most of these patients resolved during danaparoid therapy, indicating that detection of low affinity antibodies to PF4-danaparoid by immunoassay may not be an absolute contraindication for danaparoid administration. While HIT patients possess antibodies to PF4-heparin, I observed that HIT antibodies will also bind to PF4 alone adsorbed on polystyrene ELISA wells but not to soluble PF4 in the absence of heparin. Having developed a technique to affinity-purify anti-PF4-heparin HIT IgG, I provide the first estimates of the avidity of HIT IgG. HIT IgG displayed relatively high functional affinity for both PF4-heparin (Kd=7-30nM) and polystyrene adsorbed PF4 alone (Kd=20-70nM). Furthermore, agarose beads coated with PF4 alone were almost as effective as beads coated with PF4 plus heparin in depleting HIT plasmas of anti-PF4-heparin antibodies. I conclude that the HIT antibodies which bind to polystyrene adsorbed PF4 without heparin are largely the same IgG molecules that bind PF4-heparin and thus most HIT antibodies bind epitope(s) on PF4 and not epitope(s) formed by part of a PF4 molecule and part of a heparin molecule. Binding of PF4 to heparin (optimal) or polystyrene/agarose (sub-optimal) promotes recognition of this epitope. Under conditions that are more physiological and sensitive than previous studies, I observed that affinity-purified HIT IgG will cause platelet aggregation upon the addition of heparin. Platelets activated with HIT IgG increased their release and surface expression of PF4. I quantitated the binding of affinity-purified HIT 125I-IgG to platelets as they activate in a plasma milieu. Binding of the HIT IgG was dependent upon heparin and some degree of platelet activation. Blocking the platelet Fc??? receptor-II with the monoclonal antibody IV.3 did not prevent HIT IgG binding to activated platelets. I conclude that anti-PF4-heparin IgG is the only component specific to HIT plasma that is required to induce platelet aggregation. The Fab region of HIT IgG binds to PF4-heparin that is on the surface of activated platelets. I propose that only then does the Fc portion of the bound IgG activate other platelets via the Fc receptor. My data support a dynamic model of platelet activation where released PF4 enhances further antibody binding and more release.

Thesis (Ph. D.)--University of Virginia, 2008. / Title from title page. Includes bibliographical references. Also available online through Digital Dissertations.

Thesis (Ph. D.)--University of Virginia, 2008. / Title from title page. Includes bibliographical references. Also available online as viewed 8/06/2009 through Digital Dissertations.

A trombocitopenia induzida por heparina (TIH) é uma síndrome imunohematológica mediada por um anticorpo que causa ativação plaquetária na presença de heparina, induz à agregação plaquetária e pode estar associada a graves e paradoxais complicações trombóticas e morte. A freqüência de TIH nos pacientes que recebem heparina por mais de cinco dias é de 1% a 5%, e está relacionada a vários fatores. Este é um estudo pioneiro no Brasil, que objetivou avaliar aqui a freqüência de TIH nos pacientes em uso de heparina, a relação ao gênero, ao tipo de heparina e a associação do genótipo da FcRIIa de receptores plaquetários. Foram selecionados 278 pacientes das Unidades de Terapia Intensiva e Unidades Coronariana do InCor-HCFMUSP, que receberam anticoagulação por heparina não fracionada (HNF) e/ou heparina de baixo peso molecular (HBPM), por pelo menos 5 dias, e excluídas as possíveis causas conhecidas de trombocitopenia. Foi realizada a contagem plaquetária pré e pós terapia com heparina, e o teste de detecção do anticorpo anti-fator 4 plaquetário/heparina (ID-PaGIA, DiaMed; e Asserachrom®-HPIA, Stago). O estudo da genotipagem da FcRIIa de receptores plaquetários foi realizado pelo método de digestão com enzima de restrição alelo específica. A freqüência de TIH encontrada foi de 6 (2,2%), e a freqüência de trombocitopenia com a presença do anticorpo anti-fator 4 plaquetário foi de 24,3%. A análise do gênero do paciente não demonstrou correlação com a TIH, nem com a trombocitopenia e nem com o anticorpo anti-fator 4 plaquetário. As mulheres apresentaram mais trombose do que os homens. A trombocitopenia ocorreu com maior freqüência nos pacientes que utilizaram os dois tipos de heparina (HNF-HBPM) e, com menor freqüência, nos que utilizaram apenas HBPM. O genótipo da FcRIIa de receptores plaquetários não apresentou relação com o gênero, nem com a TIH. Este estudo determinou a freqüência de TIH em uma população brasileira com uso de heparina e auxiliou no diagnostico. O melhor teste para detectar o anticorpo anti-fator 4 plaquetário/heparina, na presença de trombocitopenia e trombose, foi o teste de imunoaglutinação ID-PaGIA (DiaMed). A utilização dos dois tipos de heparina promoveu uma maior freqüência de trombocitopenia. Porém, novos estudos precisam confirmar as relações entre o tipo de heparina, com a trombocitopenia e com a TIH / Heparin induced thrombocytopenia (HIT) is an immune-hematologic syndrome mediated by a heparin dependent antibody that causes platelet activation, platelet aggregation, and can be associated with thrombosis and death. HIT occurs in about 1-5% of patients receiving heparin therapy up to 5 days or more. Many factors influence on the frequency of HIT. This is a pioneer Brazilian study to determine the frequency of HIT on patients under heparin therapy, and the relationship of HIT with gender, heparin type and the FcRIIa platelet receptor genotype. 278 patients from the Intensive Care Unit and Cardiac Care Unit at InCor-HCFMUSP treated with Unfractionated Heparin (UFH) and/or Low Molecular Weight Heparin (LMWH) for 5 or more days were studied. Known causes of thrombocytopenia were excluded. Platelet count was monitored pre and post heparin therapy. All selected patients were tested for detection of anti-heparin/PF4 antibody (ID-PaGIA, DiaMed; and Asserachrom®-HPIA, Stago). HIT frequency found was 6 (2,2%) and the frequency of thrombocytopenia (determined by a decrease in the platelet count below 50%, after the introduction of heparin therapy) and positive anti-heparin/PF4 antibody test was 24,3%. Patients gender was not related to TIH, neither to thrombocytopenia nor to the presence of antiheparin/ PF4 antibody. Thrombosis events were more frequent in women than in men. Thrombocytopenia, related to the type of heparin, was more frequent in patients that had used both types of heparin and less frequent in those that used only LMWH. FcRIIa platelet receptor genotype was associated with neither HIT nor with gender. This study has provided the frequency of HIT in a Brazilian patient population under heparin therapy and auxiliary in the HIT diagnosis. The ID-PaGIA (DiaMed) was shown to be the best test to correlate the presence of anti-heparin/PF4 antibody to thrombocytopenia and thrombosis event. The use of both heparin types promotes more thrombocytopenia. New studies are needed to confirm the relationship between heparin type and thrombocytopenia with HIT

京都大学 / 0048 / 新制・論文博士 / 博士(医学) / 乙第13159号 / 論医博第2146号 / 新制||医||1029(附属図書館) / (主査)教授 河本 宏, 教授 前川 平, 教授 髙折 晃史 / 学位規則第4条第2項該当 / Doctor of Medical Science / Kyoto University / DFAM

Indiana University-Purdue University Indianapolis (IUPUI) / Solid organ transplantation is limited by available donor allografts. Pig to human transplantation, xenotransplantation, could potentially solve this problem if physiologic and immunologic incompatibilities are overcome. Genetic modifications of pigs have proven valuable in the study of xenotransplantation by improving pig to human compatibility. More genetic targets must be identified for clinical success. First, this study examines platelet homeostasis incompatibilities leading to acute thrombocytopenia in liver xenotransplantation. Mechanisms for xenogeneic thrombocytopenia were evaluated using liver macrophages, Kupffer cells, leading to identification of CD18, beta-2 integrin, as a potential target for modification. When disruption of CD18 was accomplished, human platelet binding and clearance by pig Kupffer cells was inhibited. Further, human and pig platelet surface carbohydrates were examined demonstrating significant differences in carbohydrates known to be involved with platelet homeostasis. Carbohydrate recognition domains of receptors responsible for platelet clearance Macrophage antigen complex-1 (CD11b/CD18) and Asialoglycoprotein receptor 1 in pigs were found to be different from those in humans, further supporting the involvement of platelet surface carbohydrate differences in xenogeneic thrombocytopenia. Second, immunologic incompatibilities due to antibody recognition of antigens resulting in antibody-mediated rejection were studied. Identification of relevant targets was systematically approached through evaluation of a known xenoantigenic protein fibronectin from genetically modified pigs. N-Glycolylneuraminic acid, a sialic acid not found in humans, was expressed on pig fibronectin and was identified as an antigenic epitope recognized by human IgG. These studies have provided further insight into xenogeneic thrombocytopenia and antibody-mediated rejection, and have identified potential targets to improve pig to human transplant compatibility.

Background: Bernard-Soulier syndrome (BSS) is a severe congenital bleeding disorder characterized by thrombocytopenia, thrombocytopathy and decreased platelet adhesion. BSS results from genetic alterations of the glycoprotein (GP) Ib/IX/V complex. Methods: We report on a patient demonstrating typical BSS phenotype (thrombocytopenia with giant platelets, bleeding symptoms). However, BSS was not diagnosed until he reached the age of 39 years. Results: Flow cytometry of the patient’s platelets revealed absence of GPIb/IX/V receptor surface expression. In addition, immunofluorescence analysis of patient’s platelets demonstrated very faint staining of GPIX. A novel homozygous deletion comprising 11 nucleotides starting at position 1644 of the GPIX gene was identified using molecular genetic analysis. Conclusions: The novel 11-nucleotide deletion (g.1644_1654del11) was identified as causing the bleeding disorder in the BSS patient. This homozygous deletion includes the last 4 nucleotides of the Kozak sequence as well as the start codon and the following 4 nucleotides of the coding sequence. The Kozak sequence is a region indispensable for the initiation of the protein translation process, thus preventing synthesis of functional GPIX protein in the case of deletion. / Hintergrund: Das Bernard-Soulier-Syndrom (BSS) ist eine angeborene Blutungsstörung, die mit Thrombozytopenie, Thrombozytopathie und verminderter Thrombozytenadhäsion assoziiert ist. BSS wird durch genetische Veränderungen des Glykoprotein(GP)-Ib/IX/V-Komplexes verursacht. Methoden: Wir berichten über einen Patienten mit typischem BSS-Phänotyp (Thrombozytopenie mit Riesenthrombozyten, Blutungssymptome). Dennoch wurde die Diagnose BSS erst im Alter von 39 Jahren gestellt. Ergebnisse: Die Durchflusszytometrie der Thrombozyten des Patienten ergab eine fehlende Oberflächenexpression des GPIb/IX/V-Rezeptors. Zusätzlich zeigten Immunfluoreszenz-Analysen der Thrombozyten eine nur sehr schwache Anfärbung von GPIX. In der molekulargenetischen Analyse wurde eine noch nicht bekannte homozygote Deletion von 11 Nukleotiden (beginnend an Position 1644 im GPIX-Gen) identifiziert. Schlussfolgerungen: Diese neue Deletion von 11 Nukleotiden (g.1644_1654del11) wurde als Ursache für die vermehrte Blutungsneigung bei dem BSS-Patienten identifiziert. Von der homozygoten Deletion betroffen sind die letzten 4 Nukleotide der Kozak-Sequenz sowie das Startkodon und weitere 4 Nukleotide des kodierenden Bereichs. Die Kozak-Sequenz ist unerlässlich für die Initiation der Translation in der Proteinbiosynthese, so dass die bei dem Patienten nachgewiesene Deletion die Synthese des funktionellen GPIX-Proteins verhindert. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.