CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A Fisalina E (FIS E), isolada das partes aÃreas da Physalis angulata (Solanaceae), foi avaliada em modelos de dermatite de contato induzida por 12-O-tetradecanoilforbol-13-acetato (TPA) e por oxazolona (OXA) em camundongos. A dermatite de contato irritativa aguda foi induzida pela administraÃÃo tÃpica de TPA (2,5μg/orelha) e avaliada apÃs 4h. A FIS E(0,125;0,25 e 0,5 mg/orelha,por via tÃpica) reduziu de forma significativa o edema de orelha induzido por TPA em 33,1; 38,7; 39%, respectivamente, e dexametasona(0,05 mg/orelha,por via tÃpica) reduziu em 95,2%. Nas mesmas doses, a FIS E assim como a Dexa, reduziram em 45,7; 52,5; 67,6 e 83,4 %, respectivamente, a atividade da mieloperoxidase (MPO) tecidual. FIS E (3, 10 e 30 mg/kg,por via oral) e Dexa (1mg/kg,por via oral) reduziram o edema de orelha em 34,8; 40,7, 47,3 e 46,6% respectivamente. Nas mesmas doses, a FIS E assim como a Dexa, reduziram em 39,3; 46,7 e 54,3 e 81,7 %, respectivamente, a atividade da mieloperoxidase (MPO) tecidual. A FIS E (0.5 mg/orelha) foi capaz de reduzir os nÃveis teciduais de TNF-α de 8,711.18 pg/ml (veÃculo) para 2,950.81 pg/ml e a dexametasona (0,05 mg/orelha) para 2,350.94 pg/ml, representando uma reduÃÃo de 66,2 e de 73 %, respectivamente. A administraÃÃo de Mifepristone (25 mg/kg,s.c), antagonista esteroidal, reverteu os efeitos da FIS E e da Dexa, por via oral e tÃpica, sobre a reduÃÃo do edema, atividade da mieloperoxidase e nÃveis de TNF-α. Na marcaÃÃo imunohistoquÃmica para TNF-α e NF-κB, a FIS E (0,5 mg/orelha) e a dexametasona (0,05 mg/orelha) reduziram a intensidade de marcaÃÃo do TNF-α e NF-κB, e o Mifepristone, reverteu esse efeito. A aÃÃo antiinflamatÃria da FIS E no modelo da dermatite aguda induzida por TPA foi confirmada pelos achados histopatolÃgicos, com a reduÃÃo do edema, infiltrado inflamatÃrio de mono e polimorfonucleares. Na dermatite crÃnica foi induzida pela aplicaÃÃo tÃpica de 20 Âl de uma soluÃÃo de TPA (2,5Âg/orelha) em dias alternados durante 10 dias, a FIS E, por via tÃpica, nas doses 0,125; 0,25 e 0,5 mg/orelha, reduziu, de forma significativa, o edema de orelha, em 19, 23 e 30,3%, respectivamente, enquanto a Dexa (0,05mg/orelha) reduziu em 32,6 %. FIS E (0,125;0,25 e 0,5 mg/orelha,por via tÃpica) e a Dexa(0,05mg/orelha) reduziram a atividade tecidual da MPO em 30; 38,8; 50,8 e 56,6%, respectivamente. A administraÃÃo oral da FIS E (3, 10 e 30 mg/kg), reduziu, de forma significativa, o edema de orelha crÃnico, em 8; 14,8; 17 %, respectivamente, e a Dexa (1 mg/kg) reduziu em 20,7 %.Nas mesmas doses, a FIS E e a Dexa reduziram a atividade da MPO em 43,1; 54,5 ;59,3 e 65 %,respectivamente. O modelo de dermatite de contato induzida por oxazolona (OXA, 1%/orelha), provocou uma hiperplasia epidÃrmica onde o INF-γ teve papel crucial. A FIS E, por via oral e tÃpica, reduziu de forma significativa, a hiperplasia epidÃrmica do 7Â ao 19Â dia de observaÃÃo. A FIS E (0.5 mg/orelha, por via tÃpica) diminuiu os nÃveis de IFN-γ em 43,5% e a Dexa (0.05 mg/orelha, via tÃpica) em 37,5%.Os resultados encontrados, comprovados pelo estudo histolÃgico, demonstram o efeito antiinflamatÃrio da Fisalina no modelo de dermatite de contato alÃrgica crÃnica. ConcluÃmos que a Fisalina E demonstrou atividade antiinflamatÃria, por via tÃpica e oral, nos modelos de dermatite de contato induzida por TPA e oxazolona em camundongos, possivelmente atravÃs da interaÃÃo com receptores de glicocorticÃides. / The physalin E (PHY E), isolated from the aerial parts of Physalis angulata (Solanaceae), was assessed in models of contact dermatitis induced by 13-acetate-12-o-tetradecanoyl-phorbol (TPA) and oxazolone (OXA) in mice. The irritant contact dermatitis was induced by acute topical administration of TPA (2,5 mg/ear) and evaluated after 4h. The PHY E topically applied to ear at 0,125;0,25 and 0,5 mg/ear and dexamethasone (Dexa), topically, a dose of 0,05 mg/ear, reduced significantly the edema ear induced by TPA in 33,1; 38,7; 39 and 95,2%, respectively. In the same doses, PHY E as well as Dexa, reduced by 45,7; 52,5; 67,6 and 83,4%, respectively, the activity of myeloperoxidase (MPO) in tissue. Oral administration of PHY E (3,10 and 30 mg/kg) and DEXA (1 mg/kg) decreased the ear edema by 34,8; 40,7;47,3 and 46,6% respectively. The same doses, PHY E as well as Dexa, reduced by 39,3; 46,7;54,3% and 81,7%, respectively, the activity of myeloperoxidase (MPO) tissue. The PHY E (0,5 mg/ear) was able to reduce the tissue levels of TNF-α of 8,711.18 pg/ml (vehicle) to 2,950.81 pg/ml and dexamethasone (0,05 mg/ear) to 2,350.94 pg/ml, showing a reduction of 66.2 and 73%, respectively.The administration of mifepristone (25 mg/kg,sc),a steroid antagonist, reversed the effects of PHY E and Dexa, orally and topically, on reducing the edema,myeloperoxidase activity and levels of TNF-α. In the immunohistochemical analysis for TNF-α and NF-κB, PHY E (0,5 mg/ear) and dexamethasone (0,05 mg/ear) reduced the intensity of marking the TNF-α and NF-κB, and mifepristone reversed this effect. The anti-inflammatory action of the PHY E in the model of the dermatitis TPA-induced were confirmed by histopathological findings, with the reduction of edema and inflammatory infiltration of mono and polymorphonuclear. The chronic dermatitis was induced by application of 20 Âl solution of TPA (2,5 mg/ear) was applied topically every other day for 10 days, PHY E topically applied (0,125, 0,25 and 0,5 mg/ear) reduced significantly the edema of the ear, 19; 23; 30,3%,and Dexa (0,05 mg/ear) decreased by 32,6%.In the same doses, PHY E and Dexa reduced tissue activity of MPO in 30; 38,8;50,8 and 56,6%, respectively. Oral administration. PHY E (3, 10 and 30 mg/kg) reduced significantly the ear edema chronic in 8, 14,8 and 17%, respectively, and Dexa (1 mg/kg) reduced in 20.7%. In the same doses,PHY E and Dexa reduced activity of MPO in 43.1, 54.5, 59.3 and 65% respectively The model of contact dermatitis induced by oxazolone (OXA, 1%/ear) caused an epidermal hyperplasia where the INF-γ played crucial role. PHY E oral and topical, has significantly reduced the epidermal hyperplasia of 7 th to 19 th day of observation Physalin E (0,5 mg/ear topically) decreased levels of IFN-γ in 43,5% and Dexa (0,05 mg/ear topically) to 37,5% when compared to vehicle. Our results, as evidenced by histological study, demonstrate the anti-inflammatory effect of physalin in the model of contact dermatitis allergic. In conclusion, the results obtained showed that PHY E presented anti-inflammatory activity either when was used by oral and topical route in the models of contact dermatitis TPA and Oxazolone-induced in mice, possible trought the interaction with glucocorticoids receptors.

A dermatite de contato alérgica decorre de uma reação imunológica, mediada por células T, contra um contactante em pessoas previamente sensibilizadas. O exame padrão ouro para o diagnóstico da DCA é o teste de contato (TC). Infelizmente, o TC demanda tempo, não é inócuo e tem algumas limitações. O teste de proliferação linfocitária (TPL) convencional, uma alternativa ao TC, apresenta baixa sensibilidade no diagnóstico de alergia ao cromo. A fim de aprimorar a sensibilidade o resultado do teste foi obtido pela detecção de citocinas no sobrenadante das culturas e não por timidina radiomarcada. Dezoito pacientes alérgicos ao cromo e 19 controles foram testados com o teste convencional e com as citocinas IFN-?, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 e rantes. Houve correlação entre alergia ao cromo e detecção das citocinas IFN-gama, IL-2, IL-5, IL-12 e IL-13. Dessas, os melhores resultados foram encontrados com a IL-13. O TPL pode ser utilizado como exame adicional ou alternativo no diagnóstico da DCA pelo cromo / Allergic contact dermatitis (ACD) elapses from an specific T cell immunologic reaction against an allergen in a sensitized individual. The gold standard exam to confirm ACD is the patch test (PT). However, PT demands time, has potential side efects and some limitations. The standard lymphocyte proliferation assay (LPA) has sensitivity in the diagnosis of chromium allergy. To optimize this test the results were obtained by detection of cytokines instead radiolabeled thymidine. Eighteen patients allergic to chromium and 19 controls were tested with the conventional LPA and for the following cytokines: IFN-?, IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IL-17 and rantes. Correlation between allergy to chromium and detection of IFN-gama, IL-2, IL-5, IL-12 e IL-13 was found. The best results were found with IL-13. LPA can be used as an alternative or additional test in the diagnosis of ACD by chromium.

O objetivo deste trabalho e o desenvolvimento de roupas utilizando malha de linho para uso como primeira camada em contato direto com o corpo em usuários com dermatite atópica. Roupas disponíveis para essas pessoas com pele sensível, incluindo algumas com algodão, muitas vezes provocam irritação na pele e uma malha de linho pode ser uma boa opção de uso, com base em suas reconhecidas propriedades de contato saudável ao corpo humano, principalmente em climas quentes. Protótipos de malhas consideradas pelo autor como clássicas ou oclusivas e outras como não oclusivas foram produzidas com algodão, fibras descontinuas de poliéster e linho em estrutura jérsei de malharia circular. As malhas foram confeccionadas em roupas para um teste de uso experimental em 4 crianças entre 5-8 anos de idade, sendo 2 delas saudáveis e duas com dermatite atópica, previamente avaliadas por um dermatologista. O resultado final demonstrou um bom grau de tolerância por todos os usuários quanto ao uso da malha com linho em contato direto com a pele. As malhas não oclusivas foram as que apresentaram os melhores índice de permeabilidade ao ar,atributo que pode evitar o bloqueio da transpiração insensível durante rotinas diárias, minimizando o abafamento, desconforto térmico e potencial gatilho de irritação da pele. Câmera infravermelha foi utilizada como uma ferramenta adicional na Avaliação térmica durante o uso. Os resultados sugerem que a malha com linho não oclusiva e uma opção viavel de uso pelos portadores de dermatite atópica em clima subtropical como o da cidade de São Paulo. O controle da estabilidade dimensional da malha com linho e amaciantes naturais amigáveis a pele para a lavagem domestica das roupas são desafios que devem ser equacionados em futuros desenvolvimentos / Linen (Linum usitatissimum L.) in single jersey of circular knitting machine for wearer with atopic dermatitis in subtropical climate The purpose of this work is the development of first layer linen knitted structure aiming wearers with Atopic Dermatitis (AD). Available clothing for these sensitive skin people, including some cotton garments, very often presents skin irritation and linen is an option based upon it recognized friendly contact to human body and comfortable wear during hot climates. Classic or occlusive and non-occlusive garments prototypes knitted in single jersey of circular knitting machines with cotton, polyester staple and linen fibers were designed to accomplish an experimental wear test with 4 children with 5-8 years old, 2 with AD and 2 health kids. Voluntaries showed good wear tolerance to knitted linen that presented highest air permeability index that can help to avoid blocking of insensible perspiration during daily routine, minimizing buffering effect and potential trigger of skin irritation. Infrared camera was used as an additional tool for thermal evaluation during use. Results suggest that non-occlusive knitted linen is a possible option for AD wears at subtropical climate like the city of Sao Paulo. Dimensional stability control of nonhocclusive knitted linen and natural home laundry softeners are challenges to be overcome in future developments

Abstract The purpose of the study was to get knowledge of everyday life of children (aged 0–10 years) with atopic dermatitis and their families. An ethnographic approach was used. Different data sets were collected by interviewing, observing and presenting questionnaires. The interviews (n = 24) and observations (n = 24) of the families (n = 7) took place at their homes. Children aged 5–10 years (n = 80), assisted by their parents, filled in a quality of life questionnaire. In the case of children younger than 5 years (n = 174) and families (n = 254), the quality of life questionnaire was filled in by the parents. The severity of atopic dermatitis of each participating child (n = 254) was evaluated by a physician. The professional perspective was introduced into the study in the form of discussions between dermatological nurses and dermatologists (n = 4). Triangulation of data, methods and analyses was performed. The quantitative data were analyzed by means of SPSS 11.5 for Windows. Categorical and ordinal variable were presented as frequency and percentage distributions, and medians, quartiles and arithmetic means were quoted. The interrelations between variables were presented as cross-tabulations, and Khi square test was used. The interviews were analyzed by methods of data-based content analysis. The more severe the child's atopic dermatitis was, the more the quality of life of the child and the family was impaired. The quality of life of the children was better than the quality of life of the families. The impact of the atopic dermatitis to the quality of life of the children was moderate and of the family severe. The children's quality of life was impaired most by itching and scratching. The need to participate in the treatment of the children's skin problems was the most important single factor that impaired the families' quality of life. The child's severe, long-term dermatitis and its treatment on a day-to-day basis were described as a desperate process. The knowledge that eczema is alleviated as the child grows older gave the families hope and helped them to cope. The avoidance of various foods and allergens and the opportunities to try different treatments raised hopes about the alleviation of eczema. On the other hand, however, these trials easily turned into desperate efforts to try any available alternative, which often wore out the mother or the whole family. Families did not receive adequate counselling or support when their quality of life was seriously compromised. The results of the study can be used when developing the public health care services to better meet the needs of families. / Tiivistelmä Tutkimuksen tarkoituksena on saada tietoa ihoatoopikkolasten (0–10 vuotta) ja heidän perheidensä arjesta. Tutkimuksen lähestymistapa on etnografinen. Erilaisia aineistoja kerättiin haastattelemalla, havainnoimalla ja kyselylomakkeilla. Perheiden (n = 7) haastattelut (n = 24) ja havainnoinnit (n = 24) tehtiin heidän kodeissaan. Perheiden arki heijastui haastattelutilanteisiin siten, että niissä ei aina ollut läsnä koko perhe, ja haastatteluun osallistujien toiveiden mukaan tapaamisia oli myös esimerkiksi ihotautien vuodeosastolla ja poliklinikalla. 5–10-vuotiaat lapset (n = 80) täyttivät itse tai vanhempiensa avustamana omaa elämänlaatuaan koskevan kyselylomakkeen. Alle 5-vuotiaiden lasten (n = 174) ja perheiden (n = 254) elämänlaatukyselyyn vastasivat vanhemmat. Lääkäri arvioi jokaisen tutkimukseen osallistuneen lapsen (n = 254) atooppisen ihottuman vaikeuden. Ammattilaisten näkökulman tutkimukseen toivat ihotautisairaanhoitajien ja -lääkäreiden kanssa käydyt keskustelut (n = 4). Tutkimuksessa käytetään aineisto-, menetelmä- ja analyysitriangulaatiota. Kvantitatiivinen aineisto analysoidaan SPSS 11.5 for Windows -ohjelmalla. Luokittelu- ja järjestysasteikollisia muuttujia kuvataan frekvenssi- ja %-jakaumilla, ja sijaintilukuina ovat mediaanit, kvartiilit ja aritmeettinen keskiarvo. Muuttujien yhteyksiä tarkastellaan ristiintaulukoimalla ja parametrittomana testinä on chi2-testi. Haastattelu- ja havainnointiaineisto analysoidaan aineistolähtöisellä sisällönanalyysillä. Mitä vaikeampi lapsen ihottuma oli, sitä enemmän se heikensi perheen ja lapsen elämänlaatua. Lasten elämänlaatu oli parempi kuin perheiden elämänlaatu. Atooppisen ihottuman vaikutus lasten elämänlaatuun oli kohtalainen ja perheiden elämänlaatuun huomattava. Lasten elämänlaatua heikensivät eniten kutina ja raapiminen. Lapsen hoidossa auttaminen oli tärkein perheen elämänlaatua heikentävä tekijä. Lapsen vaikeaa, pitkäaikaista ihottumaa ja sen päivästä toiseen jatkuvaa hoitamista kuvattiin ajoittain epätoivoiseksi. Toisaalta tieto, että ihottuma lievittyy lapsen kasvaessa, antoi toivoa ja auttoi perhettä jaksamaan. Perheet hankkivat ja saivat paljon ja monenlaista tietoa lapsen atooppisesta ihottumasta ja sen hoitamisesta, joten vanhemmat joutuivat suodattamaan ja puntaroimaan saamaansa tietoa oman lapsen tilanteeseen sopivaksi. Yhtäältä erilaisten ruokien tai ärsyttävien tekijöiden välttämiseen ja hoitovaihtoehtojen kokeilemiseen liittyi toivoa lapsen ihottuman lievittymisestä. Toisaalta kokeileminen saattoi kääntyä epätoivoiseksi tarttumiseksi jokaiseen tarjolla olevaan oljenkorteen, mikä puolestaan johti äidin tai koko perheen väsymiseen. Perheiden ohjauksessa ja tukemisessa oli puutteita, kun perheen elämänlaatu oli huomattavasti heikentynyt. Perheet käyttivät monipuolisesti terveydenhuollon palveluja, mutta ajoittain hoito koettiin pirstaleiseksi, ja hoitovastuu jäi perheen itsensä kannettavaksi. Tutkimustuloksia voidaan käyttää kehitettäessä terveydenhuoltopalveluja paremmin perheiden tarpeita vastaaviksi. Keskeistä on tunnistaa ne perheet, joiden elämänlaatu on huomattavasti heikentynyt lapsen atooppisen ihottuman vuoksi. Ihoatoopikkolapsen hoitaminen on moniammatillista yhteistyötä, jonka tekemistä yli ammattikunta- tai sektorirajojen on entisestään syytä tehostaa.

Les sensibilisants de contact sont des molécules réactives électrophiles qui ont la capacité de modifier des protéines de la peau pour former un antigène. Au delà de ce mécanisme d'hapténisation, le signal de danger induit par les sensibilisants conduisant à l'activation des cellules dendritiques (DC) est un élément déterminant dans l'induction de cellules T spécifiques de l'haptène. Dans le contexte du 7ième amendement à la directive cosmétique européenne, la mise en place d'une batterie de tests in vitro permettant de prédire le potentiel sensibilisant de molécules est indispensable pour l'industrie cosmétique. Tandis que la plupart des études in vitro étudient les signaux de danger induits par les sensibilisants dans des modèles homéostasiques, nous nous sommes intéressés à l'effet des sensibilisants sur la mise en place d'une réponse inflammatoire. Lorsque la lignée U937 est différenciée avec du PMA et stimulée avec du LPS, les facteurs de transcription NF-κB et Nrf2 sont activés et l'acide arachidonique (AA) est métabolisé au travers de la cascade cPLA2 / COX-2. L'ensemble de ces voies activées conduit à la production par les U937 d'un grand nombre de médiateurs inflammatoires (IL-1β, TNF-α, IL-6, IL-10, IL-8, PGE2, PGD2, TxB2). Dans ce modèle, nous avons analysé l'effet de 6 sensibilisants de potentiels variés (DNCB, PPD, HQ, PG, CIN, EUG) et montré que de façon inattendue, tous les sensibilisants étudiés diminuent significativement et de façon spécifique la production de tous les prostanoïdes et en particulier de PGE2 induite par PMA/LPS. Nous avons de plus démontré que selon les sensibilisants, les cibles de cette inhibition au sein de la cascade métabolique de l'AA diffèrent, même si elles se focalisent la plupart du temps (sauf pour le DNCB) sur l'enzyme COX-2 (inhibition de son expression et/ou de son activité). Pour le DNCB, le mécanisme d'inhibition semble plutôt impliquer sa capacité à réagir fortement avec les groupements résidus thiols, ce qui se traduit en particulier par la déplétion du GSH intracellulaire et engendrerait l'inhibition des synthases dépendantes du GSH pour leurs activités. En parallèle de cette étude mécanistique, nous avons appréhendé la problématique du point de vue statistique et vérifié sur un set plus important et diversifié de molécules (160 molécules) que le paramètre « inhibition de PGE2 » pouvait être un bon test de prédiction de l'HSRC. L'étude statistique a permis de déterminer le modèle prédictif du test PGE2 et de mettre en évidence de bonnes performances (78%) par rapport aux prédictions du LLNA. Au-delà, une certaine complémentarité du test PGE2 avec d'autres tests in vitro (MUSST, Nrf2-HTS) a pu être mise en évidence. En conclusion, au travers de cette étude, nous avons pu mettre en évidence de nouvelles propriétés biochimiques des sensibilisants. Même si la signification biologique de la diminution de PGE2 par les sensibilisants de contact demeure complexe d'interprétation, ce paramètre a permis le développement d'un test qui prédit avec de bonnes performances le caractère sensibilisant de molécules et dont la position au sein d'une batterie prédictive d'évaluation de l'allergie de contact reste à être précisée. / Contact sensitizers are defined as reactive molecules (electrophilic) which have the ability to modify skin proteins to form an antigen (hapten). In addition to the haptenation mechanism, danger signals, leading to the activation of dendritic cells, are described to be crucial for the effective induction of an hapten-specific T cell immune response. In the context of the 7th amendment to the Cosmetic Directive, the cosmetic industry is concerned by the challenge of finding non-animal approaches to assess the sensitizing potential of chemicals. While danger signals induced by sensitizers in steady-state conditions have already been analyzed, we chose to investigate the impact of sensitizers on the course of an inflammatory response. For this purpose we used the U937 cell line differentiated with PMA and activated with LPS. In these conditions, cells produce a large amount of inflammatory mediators (IL-β, TNF-α, IL-6, IL-10, IL-8, PGE2, PGD2, TxB2) through the activation of pathways leading to the activation of the transcription factors NF-κB and Nrf2 and through AA metabolism by the cPLA2/COX-2 cascade. Interestingly, we showed that 6 contact sensitizers with various potential (DNCB, PPD, HQ, PG, CIN, EUG) significally and specifically decrease the production of prostanoïds and in particular of PGE2 induced by PMA/LPS. We further demonstrated that there is no unique inhibition profile of the sensitizers even if the majority (except for DNCB) of the effects applies on COX-2 (i.e. inhibition of the expression and/or activity). For DNCB, inhibition mechanism appears to be dependant of its capacity to react with thiols residues and in particular to deplete intracellular glutathione possibly leading to the inactivation of the PG-synthases. In parallel, we assess a statistical analysis on 160 molecules that allow us to define the test parameters (a molecule is a sensitizer if the PGE2 inhibition at 24h is more than 60%) and to calculate the test performance toward LLNA (78%). Moreover we demonstrated that the PGE2 test could be complementary to other already existing in vitro tests like MUSST or Nrf2-HTS. In summary, we add here a new insight into the multiple biochemical effects described so far for sensitizers. Even if the underlying biological relevance remains unclear, the parameter “PGE2 inhibition” is good test for skin sensitization evaluation. Further studies will precise how this parameter could be implemented into an alternative testing strategy for the evaluation of skin sensitization.

The aim of the diploma thesis was to provide an overview of the prevalence of allergenic and potentially allergenic metals in eye shadows. The diploma thesis gives an overview of the legislation on cosmetics and description of the types of allergenic reactions caused by allergenic and potential allergenic metals in eye shadows. Listed below are the preparation methods of the samples for analysis and the list of the methods by which can the selected metals be analyzed. The experimental part of the diploma thesis deals with an analysis of selected allergenic and potentially allergenic metals in eye shadows. In the experimental part of diploma thesis method of sample preparation for analysis of eye shadows and a method of analysis of sample of eye shadows by ICP-MS have been optimized. Monitored analytes were selected metals. Altogether 6 samples of eye shadows from different manufacturers were chosen. Measured results have been statistically processed, confronted with the applicable legislation and assessed in terms of possible allergic reactions.

Die Mastzellen der Haut sind bedeutende Immuneffektorzellen in der Pathogenese der Caninen Atopischen Dermatitis (CAD; OLIVRY et al. 1997). Diese Zellen schütten in der Sofort- und in der Spätphase der Überempfindlichkeitsreaktion des Typs I Entzündungsmediatoren aus. Diätetisch verabreichte Fettsäuren werden in zelluläre Membranen eingebaut und sind somit in der Lage, die Produktion und Freisetzung dieser Entzündungsmediatoren zu beeinflussen. In der Praxis konnte gezeigt werden, dass eine diätetische Ergänzung von n6- und n3-Fettsäuren im Verhältnis von 5 zu 1 eine Linderung der klinischen Symptomatik bei 40% der an CAD leidenden Hunde herbeiführte (SCOTT et al. 1997). Das Ziel der vorliegenden Arbeit war es, zu überprüfen, welche Auswirkungen der Einbau supplementierter n6- und n3-Fettsäuren auf die Fettsäurenzusammensetzung und die Prostaglandinfreisetzung caniner Mastocytomzellen (C2) hat und ob diese Zellen in Bezug auf ihren Fettsäurenstoffwechsel als Modell für die CAD geeignet sind. Die Kultivierung der Zellen erfolgte in einem Grundmedium (DEH) oder in mit 14 µM Linol- (C18:2n6, DEH-LA), Gammalinolen- (C18:3n6, DEH-GLA), Arachidon- (C20:4n6, DEH-AA), a-Linolen- (C18:3n3, DEH-LnA), Eicosapentaen- (C20:5n3, DEH-EPA) oder Docosahexaensäure (C22:6n3, DEH-DHA) angereichertem Medium. Das Wachstum der C2 wurde in allen Kulturmedien über 11 Tage kontrolliert. Für die weiteren Untersuchungen wurden die Zellen am 4. bzw. 8. Tag geerntet, zweimal mit phosphatgepufferter Kochsalzlösung gewaschen und anschließend unter Stickstoff getrocknet. Die Ermittlung der Fettsäurenzusammensetzung der C2 erfolgte mittels Gaschromatographie nach Extraktion und Umesterung der Phospholipide. Dabei wurde L-a-Phosphatidylcholin-C17:0 als Interner Standard genutzt. Für die Bestimmung der Prostaglandine (PG) D2 und E2 wurden die Zellen mit dem Wespengift Mastoparan stimuliert. PGD2 wurde mittels eines PGD2-Methoxim-Enzym-Immunoassay (EIA) und PGE2 wurde mit Hilfe eines Radio-Immunassays (RIA) bestimmt. Die C2 zeigten in allen Kulturmedien eine Vermehrung lebender Zellen bis zum 8. Kultivierungstag, danach nahm die Zahl der abgestorbenen Zellen deutlich zu. Die Fettsäurensupplementierung beeinflusste das Zellwachstum nicht. Die erhöhte Zufuhr der Fettsäuren bewirkte eine Konzentrationserhöhung der entsprechenden Fettsäuren in den C2 (LA 4,9-fach, GLA 6,9-fach, AA 6-fach, LnA 9,3-fach, EPA 6,5-fach, DHA 8,4-fach). Weiterhin wurden signifikante Erhöhungen von Fettsäurenmetaboliten, die über die Elongasen und die D6-Desaturase aus den zugegebenen Fettsäuren gebildet werden, in den C2 gefunden. Produkte der D5-Desaturase waren dagegen nur in geringen Mengen nachweisbar. Ein zeitabhängiger Effekt des Einbaus der geprüften supplementierten Fettsäuren konnte nur für LA festgestellt werden, welche nach 8 Tagen in DEH-LA kultivierten C2 signifikant stärker eingebaut wurde als nach 4 Tagen. Die vorliegenden Ergebnisse lassen die Schlussfolgerung zu, dass in den C2 eine geringe Aktivität der D5-Desaturase vorliegt. Da eine niedrige Aktivität dieser Desaturase als möglicher Pathogenesemechanismus für das Auftreten der CAD verantwortlich gemacht wird, erscheinen die C2 als Modell für weitere Untersuchungen der CAD geeignet. Die durch Mastoparan stimulierte Freisetzung von PGE2 der C2 war bei der Kultivierung der Zellen im DEH-LnA und DEH-DHA signifikant erniedrigt und im DEH-AA und DEH-EPA signifikant erhöht. Die Ursache für die unterschiedlichen PGE2-Konzentrationen in C2 nach dem Zusatz der verschiedenen n3-Fettsäuren (LnA, EPA, DHA) ist bisher unklar. Verschiedene Möglichkeiten der Beeinflussung des Prostaglandinstoffwechsels durch diese Fettsäuren werden diskutiert. Auf Grund der erhaltenen Ergebnisse können die C2 als Modell genutzt werden, um die Mechanismen der Produktion von Prostaglandinen oder anderen Entzündungsmediatoren näher zu untersuchen und somit zur Erforschung der Pathogenesemechanismen der atopischen Dermatitis des Hundes sowie des Menschen beizutragen. / Cutaneous mast cells are considered as key immune effector cells in the pathogenesis of canine atopic dermatitis (CAD; OLIVRY et al. 1997). These cells release immediate-phase and late-phase mediators of inflammation. Dietary fatty acids are incorporated in cellular membranes and seem to influence mediator production and release. A dietary intervention with n6- and n3-fatty acids with a ratio from 5 to 1 alleviated clinical symptoms in 40% of atopic dogs (SCOTT et al. 1997). The purpose of this study was to examine the effects of n6- and n3-fatty acids on the fatty acid composition and the production of prostaglandins in canine mastocytoma cells (C2) as a possible model for CAD. Cells were cultured in a basic medium (DEH) or with additional 14 µM linoleic (C18:2n6, DEH-LA), gammalinolenic (C18:3n6, DEH-GLA), arachidonic (C20:4n6, DEH-AA), a-linolenic (C18:3n3, DEH-LnA), eicosapentaenoic (C20:5n3) or docosahexaenoic acid (C22:6n3, DEH-DHA). Cell growth was examined for 11 days in all media. The cells were harvested after 4 or 8 days, washed twice with phosphated buffered saline and dried under nitrogen for fatty acid analysis. The fatty acid composition was determined by gas chromatography after extraction and transesterification of the phospholipids using di-C17-phosphatidylcholin as internal standard. For measurment of prostaglandin (PG) D2 and E2 the C2 were stimulated with the wasp venom peptide mastoparan. PGD2 was measured by PGD2-methoxim-enzymimmunoassay (EIA) and PGE2 was determined by radioimmunoassay (RIA). Cell growth increased from day 1 to 8 and decreased thereafter in all media conditions. The supplied fatty acid did not influence the cell growth. Added fatty acids increased the concentration of these fatty acids in C2 (LA 4.9-fold, GLA 6.9-fold, AA 6-fold, LnA 9.3-fold, EPA 6.5-fold, DHA 8.4-fold). Futhermore elongated and D6-desaturated products of the corresponding fatty acids were significantly elevated, however D5-desaturated products were not measurable. An increased time dependent incorporation was only detectable for LA after culturing C2 in DEH-LA. The results let us assume that C2 has no activity of the D5-desaturase. If the assumed low activity of these desaturase is one of the mechanisms underlying the pathogenesis of CAD, C2 seems to be an adequate model for CAD. The production of PGE2 after stimulation with mastoparan was significantly reduced when C2 were cultured in DEH-LnA and DEH-DHA and was significantly increased when C2 were cultured in DEH-AA and DEH-EPA. The reason for the different PGE2-production in C2 after the treatment with the n3-fatty acids (LnA, EPA or DHA) being unsettled. The observed results suggest, that C2 could be used to investigate the mechanisms of production and release of prostaglandins or other mediators as a model to improve our understanding of the pathogenesis of canine or human atopic dermatitis.

Die Dermatitis digitalis (DD) ist eine weltweit verbreitete infektiöse Klauenerkrankung mit negativem Einfluss auf das Wohlbefinden und die Leistung von Milchrindern. Durch die multifaktoriell bedingte Ätiologie und die unterschiedlichen klinischen Erscheinungsformen gestaltet sich die Therapie und Prophylaxe der Erkrankung als schwierig. Schätzungen für Erblichkeiten im moderaten Bereich und die Identifizierung von verschiedenen Kuhtypen hinsichtlich der Anfälligkeit für DD unterstreichen die mögliche Beteiligung von wirtseigenen genetischen Faktoren an der Entstehung der Erkrankung. Unter Anwendung einer intensiven Phänotypisierung DD-betroffener Tiere wurden im Rahmen dieser Arbeit genetisch bedingte Hintergründe der Erkrankung untersucht, Erblichkeiten berechnet und eine Zuchtwertschätzung für DD entwickelt. Zudem war beabsichtigt über eine genomweite Assoziationsstudie (GWAS) relevante chromosomale Bereiche, Kandidatengene und funktionelle Gengruppen für Merkmale, die die Anfälligkeit und den Verlauf der Erkrankung beschreiben, zu identifizieren. Die Ergebnisse der vorliegenden Arbeit liefern neue Erkenntnisse hinsichtlich einer züchterischen sowie genomischen Bearbeitung der DD des Holstein-Rindes. Dabei stellt die Nutzung von Phänotypen aus der wiederholten Einstufung von Kühen mittels M-Stadien-Klassifizierungssystem eine Innovation gegenüber der bisherigen Zuchtwertschätzung für Klauenerkrankungen dar. Insgesamt bestätigen die Ergebnisse eine bereits angenommene polygenetische Beeinflussung der DD beim Holstein-Rind. Eine gezielte züchterische Bearbeitung sollte flächendeckend und ergänzend zu allgemeinen Präventions- und therapeutischen Maßnahmen eingesetzt werden.

A prospective study was performed to evaluate a biological medication in the treatment of digital dermatitis (DD) in dairy cattle. The study was divided into four parts; i) on farm evaluation of DD and treatment effects and comparison between the biological ointment and OTC-spray, ii) statistical evaluation, iii) histological examination using FISH and iv) microbiological examination and culture if bacteria found in biopsies from infected skin.

No description available.