Das Auftreten manifester transplantationsassoziierter thrombotischer Mikroangiopathie sowie das Auftreten leichterer, zumeist subklinischer mikroangiopathischer Veränderungen wurde in einem Kollektiv von Patienten untersucht, die zur Behandlung einer steroidrefraktären oder abhängigen akuten oder chronischen GvHD nach allogener Stammzelltransplantation eine Therapie mit extrakorporalen Photopheresen erhielten. Subklinische mikroangiopathische Schädigung wurde durch ein auf den Messwerten von Fragmentozyten, Thrombozyten sowie der LDH-Aktivität beruhendes Bewertungsschema festgestellt. Das Auftreten von ausgeprägten mikroangiopathischen Veränderungen (Schweregrad 2) ging mit einem erniedrigten Gesamtüberleben einher. Die Reduktion der Steroiddosis und die Verringerung der GvHD-Manifestationen wurden als Parameter für die Wirksamkeit der ECP analysiert. Ihre Effektivität wurde bestätigt. Trotz der nachgewiesenen Effektivität der ECP in der Kontrolle der GvHD zeigte sich eine Tendenz zur Zunahme mikroangiopathischer Veränderungen unter Therapie mit ECP, in Einzelfällen mit schwerwiegenden klinischen Konsequenzen.

<p>Granulocyte adhesion and release of their granule proteins are key steps during selective accumulation of a certain cell to an inflammatory site. Eosinophils are specifically recruited to sites of allergic inflammation and parasitic infection, whereas neutrophil influx predominates in bacterial infection and rheumatoid arthritis. </p><p>A simple, reliable and convenient method was developed for the measurement of granulocyte adhesion and release of granule proteins by using the normal population of granulocytes. The design allows simultaneous quantitative assessment of eosinophil and neutrophil adhesion to proteins and degranulation. </p><p>Using this method, manganese ions (Mn²⁺) induced a higher level of eosinophil adhesion to fibronectin, fibrinogen and albumin as compared with neutrophils. PMA induced comparable levels of eosinophil and neutrophil adhesion. F-MLP stimulated a rapid, short-term adhesion of neutrophils to fibrinogen. </p><p>In the same conditions PMA alone stimulated a dose-dependent release of ECP from cells that adhered to both fibronectin and fibrinogen. Meanwhile, Mn²⁺ amplified the release of ECP induced by PMA. Furthermore, release of ECP was shown to be associated with cell death.</p><p>PMA, in combination with Mn²⁺, induced a marked release of ~ 80%of the intracellular content of lactoferrin and HNL in neutrophils. PMA or f-MLP alone induced 30-40% release of lactoferrin and HNL. A maximal release of MPO of 15-20% was obtained from neutrophils stimulated by PMA and Mn²⁺. Release of lactoferrin and HNL showed a significant negative relationship to the viability of cells.</p><p>Stimulated by PMA, eosinophils from pollen-atopic patients during early pollen season displayed a markedly enhanced adhesion and release of ECP of eosinophils compared with eosinophils from the references. Priming with IL-5 caused a significantly higher adhesion and release of ECP by eosinophils in response to PMA. GM-CSF priming enhanced eosinophil adhesion in response to PAF and PMA plus Mn²⁺, but did not enhance the release of ECP.</p><p>In conclusion, the assay allows a simple quantification of eosinophil and neutrophil adhesion, as well as degranulation by using the normal population of granulocytes. Cellular adhesion plays an important role in the regulation of both eosinophil and neutrophil degranulation, but adhesion and degranulation can be induced separately.</p>

Granulocyte adhesion and release of their granule proteins are key steps during selective accumulation of a certain cell to an inflammatory site. Eosinophils are specifically recruited to sites of allergic inflammation and parasitic infection, whereas neutrophil influx predominates in bacterial infection and rheumatoid arthritis. A simple, reliable and convenient method was developed for the measurement of granulocyte adhesion and release of granule proteins by using the normal population of granulocytes. The design allows simultaneous quantitative assessment of eosinophil and neutrophil adhesion to proteins and degranulation. Using this method, manganese ions (Mn2+) induced a higher level of eosinophil adhesion to fibronectin, fibrinogen and albumin as compared with neutrophils. PMA induced comparable levels of eosinophil and neutrophil adhesion. F-MLP stimulated a rapid, short-term adhesion of neutrophils to fibrinogen. In the same conditions PMA alone stimulated a dose-dependent release of ECP from cells that adhered to both fibronectin and fibrinogen. Meanwhile, Mn2+ amplified the release of ECP induced by PMA. Furthermore, release of ECP was shown to be associated with cell death. PMA, in combination with Mn2+, induced a marked release of ~ 80%of the intracellular content of lactoferrin and HNL in neutrophils. PMA or f-MLP alone induced 30-40% release of lactoferrin and HNL. A maximal release of MPO of 15-20% was obtained from neutrophils stimulated by PMA and Mn2+. Release of lactoferrin and HNL showed a significant negative relationship to the viability of cells. Stimulated by PMA, eosinophils from pollen-atopic patients during early pollen season displayed a markedly enhanced adhesion and release of ECP of eosinophils compared with eosinophils from the references. Priming with IL-5 caused a significantly higher adhesion and release of ECP by eosinophils in response to PMA. GM-CSF priming enhanced eosinophil adhesion in response to PAF and PMA plus Mn2+, but did not enhance the release of ECP. In conclusion, the assay allows a simple quantification of eosinophil and neutrophil adhesion, as well as degranulation by using the normal population of granulocytes. Cellular adhesion plays an important role in the regulation of both eosinophil and neutrophil degranulation, but adhesion and degranulation can be induced separately.

Submitted by GABRIEL MAKSOUD GRECO null (gabrielmaksoud@gmail.com) on 2016-08-01T16:57:15Z No. of bitstreams: 1 Gabriel Maksoud Greco - Tese de Doutorado.pdf: 942153 bytes, checksum: 3897e01a5d02d842516f84f5662d1905 (MD5) / Rejected by Ana Paula Grisoto (grisotoana@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: O arquivo submetido está sem a ficha catalográfica. A versão submetida por você é considerada a versão final da dissertação/tese, portanto não poderá ocorrer qualquer alteração em seu conteúdo após a aprovação. Corrija esta informação e realize uma nova submissão contendo o arquivo correto. Agradecemos a compreensão. on 2016-08-02T19:54:49Z (GMT) / Submitted by GABRIEL MAKSOUD GRECO null (gabrielmaksoud@gmail.com) on 2016-08-05T20:05:46Z No. of bitstreams: 1 Gabriel Maksoud Greco - Tese de Doutorado com Ficha Catalográfica.pdf: 1246379 bytes, checksum: abd353a17deff1c72be3554b66dfb1c3 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-08-09T16:43:54Z (GMT) No. of bitstreams: 1 greco_gm_dr_bot.pdf: 1246379 bytes, checksum: abd353a17deff1c72be3554b66dfb1c3 (MD5) / Made available in DSpace on 2016-08-09T16:43:54Z (GMT). No. of bitstreams: 1 greco_gm_dr_bot.pdf: 1246379 bytes, checksum: abd353a17deff1c72be3554b66dfb1c3 (MD5) Previous issue date: 2016-05-31 / A manipulação farmacológica do ciclo estral é rotineiramente utilizada na égua, pelo fato desta apresentar grande variação na duração do estro e no intervalo entre o seu início e a ovulação. Como maiores taxas de concepção decorrem da deposição do sêmen no trato reprodutivo feminino próximo à ovulação, diversos agentes foram empregados objetivando sua indução e sincronização, em especial a gonadotrofina coriônica humana (hCG) e o acetato de deslorelina. O mecanismo de ação destes fármacos envolve o hormônio luteinizante (LH), sendo que o hCG mimetiza suas ações e o acetato de deslorelina estimula sua liberação pela adenohipófise. Incremento semelhante às concentrações circulantes de LH tem sido observado em vacas com o uso de cipionato de estradiol (ECP), o qual é usualmente empregado na indução e sincronização de ovulações na espécie pelo seu baixo custo. A fim de auxiliar na escolha do protocolo ideal para a indução de ovulação em éguas, o presente estudo objetivou avaliar o efeito do tratamento com o hCG e/ou com o acetato de deslorelina, nas doses preconizadas ou em subdoses, sobre as concentrações plasmáticas de LH, a indução e a sincronização de ovulação, o grau de edema endometrial, o diâmetro folicular e a sua taxa de desenvolvimento. Ainda, buscou-se verificar o resultado da aplicação de ECP sobre a indução e sincronização da ovulação e as concentrações plasmáticas de LH, bem como a existência de uma correlação entre a taxa de crescimento, o diâmetro folicular e o edema endometrial sobre a fertilidade pós-indução com a dose preconizada de hCG. Os resultados obtidos através do presente trabalho nos permitiram concluir que: a) os protocolos propostos com o hCG e/ou o DES induziram e sincronizaram a ovulação de forma eficaz, estimulando sua ocorrência de folículos pré-ovulatórios menores; b) o DES estimulou maior liberação de LH que o hCG após 24 horas; c) a adição do hCG ao tratamento com DES não promoveu um incremento nas concentrações circulantes de LH, seja nas doses preconizadas ou em subdoses; d) o tratamento com ECP não resultou em aumento nas concentrações circulantes de LH, tendo inibido sua liberação em alguns animais, pré-dispondo ao surgimento de folículos anovulatórios e atresia folicular; e) o uso de subdoses de hCG e/ou DES levaram a um maior incremento nas concentrações circulantes de LH após 48 horas, por um provável mecanismo de “down-regulation” que ocorre após o uso das doses recomendadas destes hormônios; f) os tratamentos com o hCG e/ou o DES não impediram o desenvolvimento folicular ou reduziram o diâmetro folicular após 24 horas; g) a indução de ovulação levou a uma redução no grau de edema endometrial após 24 horas; o qual persistiu à ovulação; h) o grau máximo de edema endometrial, bem como seu escore à detecção da ovulação não influenciaram a fertilidade pós-indução com 2.000 UI de hCG; i) o tamanho e a taxa de desenvolvimento folicular pós-indução com 2.000 UI de hCG tenderam a uma correlação positiva com a taxa de fertilidade pós-inseminação artificial. / Pharmacological manipulation of the equine estrous cycle is routinely applied, due to the fact that mares show great disparity on estrous duration and in the interval between its onset and ovulation. Since higher pregnancy rates are obtained when semen is deposited in the female reproductive tract at a time close to ovulation, several ovulation inducting agents have been used, specially human chorionic gonadotropin (hCG) and deslorelin acetate (DES). The mechanism of action of these drugs involves the luteinizing hormone (LH), for hCG mimics its actions and DES stimulates endogenous adenohypophyseal secretion. Similar increment in circulating LH concentrations has been observed in cows treated with estradiol cypionate (ECP), which is usually administrated as an interesting and cheaper alternative to induce and synchronize ovulations in the bovine species. In order to aid recognition of the ideal ovulation induction and synchronization protocol in the mare, the present study evaluated the effect of treatment with hCG and/or DES, using the recommended or a subdose, on ovulation induction and synchronization, plasmatic LH concentrations, grade of endometrial edema, follicular growth and diameter. Thus, this experiment intended to verify the effect of ECP administration on ovulation induction, synchronization and plasmatic LH concentrations, as well as the existence of a correlation between follicular growth, pre-ovulatory diameter and endometrial edema with fertility of mares induced using the recommended hCG dose. The obtained results led to the following conclusions: a) protocols using hCG and/or DES successfully induced and synchronized ovulations, stimulating smaller pre-ovulatory follicles to ovulate; b) DES stimulated a higher LH secretion than hCG 24 hours post-injection; c) combining hCG to DES treatment did not favor LH secretion; d) estradiol cypionate treatment did not induce ovulation or favor plasmatic LH concentrations, but, instead, lead to a substantial anovulation and follicular atresia rate, possibly due to its decrease in some animals; e) subdoses of hCG and DES stimulated a higher LH increase after 48 hours, possibly due to a down-regulation mechanism upon administration of the recommended dose; f) treatment with hCG and/or DES did not impair follicular development after 24 hours; g) ovulation induction decreased endometrial edema after 24 hours, which persisted at ovulation; h) fertility of mares treated with 2,000 IU of hCG was not influenced by the maximum endometrial edema score observed during estrous or the grade of edema the day an ovulation was detected; i) follicle size and growth rate post-induction with 2,000 IU of hCG tend to bear a positive correlation with fertility.

The objective of this work is to evaluate, through the Environmental-SCP model, the strategies adopted by a Brazilian textile company. In this context, it shows a longitudinal analysis of environmental strategies adopted by the company for eleven years, from 1996 to 2006. The research was based on an environmental strategy evaluation model, called Environmental-SCP model, proposed by Abreu (2001). The model links environmental performance to the conducts of the companies exposed to the pressure of the industry in which they operate. Thus, it was identified the environmental pressures, conducts and performance of the company in the last eleven years. Finally, this work presents the companyÂs strategic positioning, in the last eleven years, face to environmental questions. This strategic positioning is determined from the matrix that clusters companies depending on their environmental behavior and industry structure pressure according to Environmental-SCP. The analyzed company is a large scale textile firm in Cearà state that maintains an environmental management system certified according to ISO 14001 since 2001. The data were collected through document analysis and structured interviews with employees. The data were organized in electronic spreadsheets for qualitative and quantitative analysis. From this analysis, three stages of environmental strategy were identified in this period. The first stage management was unaware of environment-related pressures and merely adopted environmental conducts with the aim of acquiring obligatory environmental license. In the second stage, the perception of external though as yet unthreatening pressures led to the implementation of an environmental management system. In the third stage, as response to increased environmental pressures threatening the companyÂs competitiveness and in consideration to demands expressed by stakeholders, management adopted environmental conducts while making processes more efficient and created a competitive advantage. This study showed the companyÂs change from relative indifference to an environmentally responsible attitude harmonizing pollution prevention and increased process efficiency. / O trabalho tem por objetivo avaliar, à luz do modelo ECP-Ambiental, as estratÃgias adotadas por uma empresa tÃxtil. Neste contexto, apresenta uma anÃlise longitudinal das estratÃgias ambientais adotadas por uma empresa tÃxtil ao longo de onze anos, compreendidos entre o perÃodo de 1996 a 2006. A pesquisa foi elaborada a partir do modelo de avaliaÃÃo da estratÃgia ambiental ECPAmbiental, proposto por Abreu (2001). Com base neste modelo, a partir das pressÃes da estrutura da indÃstria, a empresa reage adotando condutas ambientais que por sua vez, afetam sua performance ambiental. Dentro deste contexto, foram identificadas as pressÃes, condutas e performance ambiental da empresa e a partir desta, determinados os Ãndices e a evoluÃÃo da pressÃo, da conduta e da performance para os Ãltimos onze anos. Por fim, o trabalho apresenta o posicionamento estratÃgico adotado pela empresa, nos Ãltimos onze anos, frente Ãs questÃes ambientais. Este posicionamento estratÃgico à determinado a partir da matriz que relaciona a pressÃo da estrutura da indÃstria com a conduta ambiental, desenvolvida a partir do modelo ECP-Ambiental. A empresa objeto deste estudo à uma empresa tÃxtil de grande porte, que possui significativa representatividade na economia do Estado do Cearà e mantÃm um sistema de gestÃo ambiental certificado na norma ISO 14001 desde 2001. A coleta de dados foi realizada por meio de anÃlise documental e aplicaÃÃo de um instrumento de coleta de dados atravÃs de entrevistas com funcionÃrios da empresa. Os dados coletados foram tabulados em planilhas eletrÃnicas, nas quais foram realizadas anÃlises qualitativa e quantitativa. A partir destas anÃlises, constatou-se que a empresa ao longo desses onze anos, passou por trÃs estÃgios: o primeiro foi caracterizado pela nÃo percepÃÃo das pressÃes ambientais, desta forma a conduta ambiental adotada era voltada ao atendimento aos requisitos do licenciamento ambiental; o segundo foi aquele em que as pressÃes ambientais foram percebidas de forma a nÃo oferecer risco à competitividade da empresa, as condutas ambientais neste estÃgio estavam voltadas à implantaÃÃo de um sistema de gestÃo ambiental; o terceiro estÃgio foi caracterizado pela percepÃÃo por parte da empresa de uma pressÃo ambiental que oferecia risco à sua competitividade, como resposta a empresa adotou condutas ambientais que atendessem as necessidades das partes interessadas e melhorassem a eficiÃncia dos processos, de forma a gerar vantagem competitiva frente aos concorrentes. No tocante à estratÃgia ambiental, a pesquisa identificou uma evoluÃÃo do posicionamento estratÃgico, migrando de uma situaÃÃo de indiferenÃa frente Ãs questÃes ambientais, para uma atuaÃÃo responsÃvel, na qual aliou prevenÃÃo da poluiÃÃo à melhoria na eficiÃncia dos processos.

Magister Educationis - MEd / The purpose of this study is to determine the nature of support services offered to Extended Curriculum Programme students in a South African university. The primary goals of support services in higher education are to support students holistically and reduce barriers to learning in the teaching and learning environment. One of the faculties in a South African university established a support unit to assist with the low throughput level. The academic support unit is housed in the Academic Development Department (ADD) in a faculty. The unit attempts to address the needs of underachieving students in the Extended Curriculum Programme (ECP). The thesis first identifies the challenges the ECP students are experiencing. Then, support services in the university and in the support unit are described. Thereafter, the challenges experienced by the centre of support services in the university and the support unit are illuminated. Qualitative data was gathered through individual interviews with senior management. Then, a focus group discussion with tutors who volunteer in a support unit was facilitated and lastly the staff members employed in the support unit were also interviewed individually. The thesis was able to identify the intrinsic and extrinsic barriers to learning the ECP students are experiencing. It became evident that the support services available in the university and the support unit are not sufficient to address the needs of the students. The challenges the support service centre of the university and the support unit are experiencing are twofold. The first is a lack of organisational resources that hinders service delivery, the second is a lack of skills and expertise in attain structures that limits the provision of support services.

Les bactéries pathogènes opportunistes (BPO) sont responsables d'une grande part de la pathologie infectieuse bactérienne. Les BPO d'origine environnementale doivent subir des changements profonds de mode de vie pour s'adapter et coloniser l'homme. Comprendre les conditions de cette adaptation permettra de préciser la notion d'opportunisme infectieux et le rôle des BPO environnementales dans l'émergence des pathogènes.Les genres Ochrobactrum et Agrobacterium regroupent des bactéries présentant une grande variété de modes de vie et établissant différentes relations avec la cellule eucaryote. Ces bactéries connues pour vivre dans l'environnement sont par ailleurs des pathogènes opportunistes de l'homme principalement responsables d'infections chez les individus immunodéprimés. Dans le cadre de ce travail nous avons entrepris une étude populationnelle par une approche de génétique multilocus sur des collections de souches cliniques et environnementales de différentes origines géographiques. Les structures de population obtenues ont été confrontées à divers caractères phénotypiques reliés à la virulence et/ou l'adaptation chez l'homme, la température de croissance, la formation de biofilm et la virulence vis-à-vis des modèles Caenorhabditis elegans et macrophages humains.Ochrobactrum anthropi et Ochrobactrum intermedium sont les deux principales espèces d'intérêt médical du genre Ochrobactrum. La population d'O. anthropi est de type épidémique qui s'organise en deux complexes clonaux (CCs). Si le CC1 regroupe à la fois des souches de diverses origines, le CC4 ne contient que des souches cliniques. Cette sous-population apparait associée à l'homme même si les caractères phénotypiques étudiés ne révèlent pas de différences entre ces deux sous populations. De la même façon, ces deux CCs ne se distinguent pas par leur comportement en modèle macrophage ou par leur diversité génomique. O. intermedium, tout comme O. anthropi, présente une forte diversité génétique toutefois, aucun regroupement des souches en fonction de leur origine n'est mis en évidence pour cette espèce. La diversité des souches cliniques apparait aussi importante que celle de l'ensemble de la population. Plusieurs arguments suggèrent une niche étroite pour cette espèce, notamment une faible diversité génomique. Par ailleurs, le faible nombre de souches environnementales associé à une meilleure croissance planctonique à 37°C qu'à 25°C et 30°C suggèrent que l'homme pourrait constituer cette niche. L'étude de la virulence d'O. intermedium en modèle macrophage ou C. elegans met en évidence différents comportements, pour autant ceux-ci ne semblent pas liés à la structure de population. Certaines souches sont capables de se multiplier dans le modèle macrophage.L'étude du genre Agrobacterium par une approche multilocus sur une collection représentative des différents modes de vie de ces bactéries met en évidence, tout comme pour O. anthropi, une sous population clinique qui regroupe près de 80% des souches de cette origine. D'autres arguments tels que la croissance à 42°C confirment que le génovar A7 peut correspondre à une sous-population associée à l'homme. Les données obtenues seront confrontées aux connaissances sur d'autres bactéries pathogènes opportunistes d'origine environnementale comme Pseudomonas aeruginosa, Stenotrophomonas maltophilia et les bactéries du complexe Burkholderia cepacia qui présentent également des sous populations associées à l'homme et/ou à certaines pathologies humaines. L'existence de ces sous populations suggère une spécialisation qui sera discutée dans le contexte de la spéciation des bactéries pathogènes afin de revisiter le concept d'opportunisme infectieux. / The opportunistic bacterial pathogens (OBP) cause the main part of bacterial infectious diseases. Environmental-borne OBP should encounter dramatic changes in lifestyle in order to colonize human beings. The conditions of this adaptation should precise concepts about OBP and emerging pathogens.The genera Ochrobactrum and Agrobacterium groups bacteria with versatile lifestyles that establish diverse relationships with the eukaryotic cells. These environmental-borne OBP caused diverse infectious diseases in immune-compromised patients. In this study, we undertook an approach of multilocus genetic on large population of environmental and clinical strains of Ochrobactrum and Agrobacterium. The population structures were compared to phenotypic traits related to adaptation and virulence in man, such as growth temperature, biofilm formation and virulence tested in Caenorhabditis elegans and human macrophages models.Ochrobactrum anthropi and Ochrobactrum intermedium are the two main Ochrobactrum species to be involved in human diseases. O. anthropi displays an epidemic population structure organized in two large clonal complexes (CCs). CC4 groups only human associated strains whereas CC1 contain environmental and clinical strains. Population genetics suggested that CC4 is a human-associated clone although phenotypic, genomic and virulence traits do not differ between CC1 and CC4 strains.As O. anthropi, O. intermedium displays a high genetic diversity without correlation between the genetic structure and the origin of strains. The level of genetic diversity among clinical strains appears as high as observed in the whole population. Several data such as a low level of genomic diversity suggested that O. intermedium is associated to a narrow ecological niche. The low number of environmental strains described for this species as well as an optimal growth at 37°C suggested that human beings could be the main niche for O. intermedium. Virulence in macrophage and C. elegans models showed diverse behaviour whereas some strains are able to survive and multiply in macrophages model.Multilocus genetics in a population of Agrobacterium spp. that displays diverse lifestyles, revealed a human associated population as observed for O. anthropi. The clinical genovar A7 groups 80% of the clinical strains included in the study, this strains growing at 42°C. Data obtained in this study will be confronted to the knowledge about other environmental-borne OBP such as Pseudomonas aeruginosa, Stenotrophomonas maltophilia and bacteria belonging to the species complex Burkholderia cepacia. All these bacteria displayed sub-populations associated to man or to a particular human disease. These sub-populations suggest a specialization process that will be described in the context of the speciation of bacterial pathogen in order to revisite the concept of « opportunisme infectieux ».

<p>Hodgkin lymphoma (HL) is characterised histologically by a minority of malignant Hodgkin and Reed-Sternberg (HRS) cells surrounded by benign cells, and clinically by a relatively good prognosis. The treatment, however, leads to a risk of serious side effects. Knowledge about the biology of the disease, particularly the interaction between the HRS cells and the surrounding cells, is essential in order to improve diagnosis and treatment. </p><p>HL patients with abundant eosinophils in the tumours have a poor prognosis, therefore the eosinophil derived protein eosinophil cationic protein (ECP) was studied. Serum-ECP (S-ECP) was elevated in most HL patients. It correlated to number of tumour eosinophils, nodular sclerosis (NS) histology, and the negative prognostic factors high erythrocyte sedimentation rate (ESR) and blood leukocyte count (WBC). A polymorphism in the ECP gene (434(G>C)) was identified and the 434GG genotype correlated to NS histology and high ESR.</p><p>The poor prognosis in patients with abundant eosinophils in the tumours has been proposed to depend on HRS cell stimulation by the eosinophils via a CD30 ligand (CD30L)-CD30 interaction. However, CD30L mRNA and protein were detected in mast cells and the predominant CD30L expressing cell in HL is the mast cell. Mast cells were shown to stimulate HRS cell lines via CD30L-CD30 interaction. The number of mast cells in HL tumours correlated to worse relapse-free survival, NS histology, high WBC, and low blood haemoglobin. </p><p>Survival in patients with early and intermediate stage HL, diagnosed between 1985 and 1992, was generally favourable and comparatively limited treatment was sufficient to produce acceptable results for most stages. The majority of relapses could be salvaged. Patients treated with a short course of chemotherapy and radiotherapy had an excellent outcome.</p><p>In conclusion prognosis is favourable in early and intermediate stages and there are possibilities for further improvements based on the fact that mast cells and eosinophils affect the biology and prognosis of HL.</p>

Hodgkin lymphoma (HL) is characterised histologically by a minority of malignant Hodgkin and Reed-Sternberg (HRS) cells surrounded by benign cells, and clinically by a relatively good prognosis. The treatment, however, leads to a risk of serious side effects. Knowledge about the biology of the disease, particularly the interaction between the HRS cells and the surrounding cells, is essential in order to improve diagnosis and treatment. HL patients with abundant eosinophils in the tumours have a poor prognosis, therefore the eosinophil derived protein eosinophil cationic protein (ECP) was studied. Serum-ECP (S-ECP) was elevated in most HL patients. It correlated to number of tumour eosinophils, nodular sclerosis (NS) histology, and the negative prognostic factors high erythrocyte sedimentation rate (ESR) and blood leukocyte count (WBC). A polymorphism in the ECP gene (434(G&gt;C)) was identified and the 434GG genotype correlated to NS histology and high ESR. The poor prognosis in patients with abundant eosinophils in the tumours has been proposed to depend on HRS cell stimulation by the eosinophils via a CD30 ligand (CD30L)-CD30 interaction. However, CD30L mRNA and protein were detected in mast cells and the predominant CD30L expressing cell in HL is the mast cell. Mast cells were shown to stimulate HRS cell lines via CD30L-CD30 interaction. The number of mast cells in HL tumours correlated to worse relapse-free survival, NS histology, high WBC, and low blood haemoglobin. Survival in patients with early and intermediate stage HL, diagnosed between 1985 and 1992, was generally favourable and comparatively limited treatment was sufficient to produce acceptable results for most stages. The majority of relapses could be salvaged. Patients treated with a short course of chemotherapy and radiotherapy had an excellent outcome. In conclusion prognosis is favourable in early and intermediate stages and there are possibilities for further improvements based on the fact that mast cells and eosinophils affect the biology and prognosis of HL.

<p>Serum levels of eosinophil granule proteins namely ECP, EPO and EPX, which are stored in the matrix of specific granules, were shown to correlate with the course of disease in disorders involving eosinophils. The concentration of eosinophil proteins in serum is the result of their release <i>in vivo</i> and <i>ex vivo</i> during the sampling procedure. Generally, eosinophils release the content of their specific granules in three ways: exocytosis, piecemeal degranulation (PM) or cytolysis. Which of them is operating in circulating eosinophils has not yet been defined. The aim of this thesis was to study the mechanisms of granule protein release from blood eosinophils in respect of protein subcellular localization and cell ultrastructure.</p><p>In patients with bacterial infections, serum levels of ECP but not EPO increased, while in patients with viral infections both proteins remained within the range of healthy controls. G-CSF is a cytokine involved in the response mechanism to bacterial but not viral infections. Administration of G-CSF to healthy subjects induced an elevation of eosinophil numbers and a preferential increase of serum EPX and ECP in comparison to EPO.</p><p>The model of PM consists of the stepwise transportation of specific granule contents from the granules towards the plasma membrane. We observed that administration of G-CSF to healthy subjects and the allergen exposure of allergic subjects during the pollen season, caused changes in the ultrastructure of eosinophil specific granules such as loosening of the matrix, granule matrix lucency and ragged losses of their core. Similar alterations of morphology had been previously described for eosinophils undergoing PM.</p><p>ECP, EPX and EPO were localized not only in the specific granules but also in extra-granular compartments as shown both by immuno electron microscopy and subcelular fractionations, An extra-granular EPX compartment was present in healthy as well as in allergic and in hypereosinophilic subjects, and there were no significant differences in its size between the groups. The size of the extra-granular compartments of ECP and EPO was increased in allergics during the season, and these compartments were clearly separate from that of EPX. Results of this show the differential mobilization ofgranule proteins in blood stream eosinophils serum and indicates PM as its mechanism.</p>