Diss. (sammanfattning) Göteborg : Univ. , 2010.

Thesis (Ph.D. in Clinical Sciences) -- University of Colorado at Denver and Health Sciences Center, 2005. / Typescript. Includes bibliographical references (leaves 140-156). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed June 30, 2007). Includes bibliographical references (p. 94-101).

Orientador: José Corrêa de Lacerda Neto / Banca: Raquel Micarelli Albernaz / Banca: Gustavo Garkalns de Souza Oliveira / Banca: Kamila Gravena / Banca: Daniela Gomes da Silva / Resumo: O grande desafio do sistema de defesa respiratório é a manutenção dos animais em baias. Pois, devido a diversos fatores, acaba por aumentar as chances do desenvolvimento de afecções respiratórias e a qualidade do material utilizado como cama é fator agravante. Diante disto, três grupos com 5 equinos cada foram submetidos a 45 dias sob o mesmo manejo, sendo dois grupos estabulados com diferentes tipos de cama, um com maravalha esterilizada (ME) e o outro com maravalha não esterilizada (MNE), e o terceiro grupo a pasto (Pasto). Foram realizadas análises do lavado broncoalveolar (LBA) (celularidade e marcadores de estresse oxidativo) e hemograma antes do início do manejo (Basal) e posteriormente a cada 15 dias (M15, M30 e M45). Também foi avaliada a presença de gêneros fúngicos nas amostras de cama, e do feno utilizado na alimentação dos animais. Os hemogramas permaneceram dentro dos valores da normalidade. Os valores obtidos de malondialdeído (MDA) e ácido úrico no LBA não apresentaram diferença entre os momentos e grupos avaliados. A vitamina C no LBA apresentou queda em seus valores em M30. Os três grupos apresentaram queda nas concentrações de glutationa reduzida em M30, tendo havido diferença significativas entre os grupos Pasto e MNE. Os grupos ME e MNE apresentaram queda nos valores de glutationa oxidada em M45, já o grupo Pasto apresentou queda constante a partir de M15, com diferença significativa em relação ao grupo ME em M30. A superóxido dismutase a... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The great challenge of the respiratory defense system is the maintenance of animals in stalls, which due to several factors ends up increasing the chances of development of respiratory diseases. The quality of the material used as bed is an aggravating factor. Three groups of 5 horses were submitted to 45 days under the same management, and two groups were housed with different types of beds, one with sterilized wood shaving (ME) and the other with unsterilized wood shaving (MNE), and the third group was maintened in the pasture. Bronchoalveolar lavage fluid (BALF) and hemoglobin analyzes were performed before baseline and then every 15 days (M15, M30 and M45). It was also evaluated the possible presence of fungal genera in bed samples, and in hay used in animal feeding. The hemograms remained within normal values. The values of malondialdehyde (MDA) and uric acid in BALF showed no difference between the moments and groups evaluated. Vitamin C in BALF showed a decrease in M30 values. The three groups showed a decrease in the concentrations of glutathione reduced in M30, and there were significant differences between the groups pasture and MNE. The ME and MNE groups showed a decrease in the values of oxidized glutathione in M45, whereas the pasture group presented a constant drop from M15, with a significant difference in relation to the ME group in M30. Superoxide dismutase increased in M30 in the MNE group, leading to a significant difference in relation to... (Complete abstract click electronic access below) / Doutor

Thesis (MTech (Tourism and Hospitality Management))--Cape Peninsula University of Technology, 2006. / The increase in popularity of fish, coupled with technological advances in the fishing industry and changes in the control and management of fishing resources, has led to a significant increase of fish processing workers from 13 million in 1970 to 38 million in 2002. Whereas allergic reactions to fish proteins were previously only documented in consumers, increasing reports of occupational fish allergies of fish processing workers has become evident. In South Africa, the reported prevalence of occupational asthma associated with fish processing workers is 2 - 8%, and the prevalence of occupational protein contact dermatitis 3 - 11%. Pilchard is one of the most consumed fish species in South Africa and the immunological analysis of this species will therefore contribute to the provision of occupational health services in the pilchard processing industry. Proteins extracted from fresh, frozen and canned pilchard (Sardinops sagax), as well as fresh samples of six other processed and consumed South African fish were characterised by denaturing protein electrophoresis and immunoblotted with different monoclonal and polyclonal antibodies. Sera from sensitised workers were subsequently used to characterise the membrane-bound pilchard proteins and analysed for human Immunoglobulin G (lgG) and Immunoglobulin E (IgE) antibodies to determine antigen recognition. A protein of 12 kDa molecular weight was found to be present in all fish protein extracts, however, at various concentrations. With the aid of the monoclonal and polyclonal antibodies, the 12 kDa protein was postulated to be parvalbumin, a known allergen in some fish species. Immunoblotting experiments for the identification of workers' IgG- and IgE-reactivities to fresh, frozen and canned pilchard showed a 12 kDa protein as an immunolgically reactive fish protein. This protein was also found to occur in dimeric, trimeric and tetrameric forms, which may have significant implications in the diagnosis and management of occupational sensitisation to pilchard.

A polinose é uma rinite alérgica sazonal que acontece pela sensibilização por pólens. Possui periodicidade anual, repetindo-se os sintomas sempre na mesma época do ano. Clinicamente, é caracterizada por rinoconjuntivite e/ou asma brônquica. A imunoterapia alérgeno-específica é o único tratamento capaz de modificar a evolução natural da doença, porém, depende fundamentalmente da correta identificação do alérgeno responsável. Diante disso e do número de pacientes que procuram o Ambulatório de Alergia da Universidade de Fortaleza com manifestações alérgicas exacerbadas na época de floração do cajueiro, o objetivo deste estudo foi produzir um extrato protéico a partir do pólen do cajueiro e identificar os alérgenos ainda não estudados presentes neste pólen. Doze pacientes residentes em Fortaleza, Nordeste do país, foram selecionados com base na história de rinite alérgica persistente e agravamento dos sintomas no momento da floração do cajueiro. Foi selecionado outro grupo com rinite alérgica que vive na mesma região, entretanto não apresenta relação clínica com a época de floração. Além disso, foram incluídos 5 indivíduos não-atópicos e expostos ao cajueiro como grupo controle. O soro desses pacientes foi testado em Western Blot 1D e 2D (WB) e as proteínas selecionadas foram submetidas à espectrometria de massas para identificação. Os epitopos foram preditos in silico pesquisando sequências detectadas por massa contra bases de dados de epítopos já conhecidos. Foi possível identificar alguns homólogos de alérgenos de outros pólens, como isoflavona redutase (Bet v 6), beta-1,3-glucanase (Ole e 9), proteína de choque térmico 70kDa (Cor a 10), além de outras proteínas que podem representar novos alergénios, tais como aminociclase, glutamina sintetase, fosfoglucomutase, ?-1,4-glucano-proteína-sintase, factor de alongamento 2 e biotina carboxilase, entre outros. A predição de epítopos revelou a possibilidade de reatividade cruzada com outros alérgenos de pólen conhecidos, tais como Phl p 4, Mal d 1, além de outros aeroalérgenos que também apareceram. Esta é a primeira descrição da alergia ao pólen do caju mostrando a reatividade específica de IgE no soros dos pacientes. A caracterização imunológica e estrutural de novos alérgenos, além de auxiliar no diagnóstico e tratamento de alergias não descritas, oferece ferramentas para prever epítopos e produzir moléculas hipoalergênicas nesta era da medicina de precisão / Pollinosis is a seasonal allergic rhinitis that develops due to pollens sensitization. Symptoms are manifested always in the same period of the year. Clinically, it is characterized by rhinoconjunctivitis and/or asthma. Allergen-specific immunotherapy is the only available treatment that can modify the natural course of the disease, however, it relies on the correct identification of the triggering allergen. Considering this and the number of patients attending the Allergy Clinic at University of Fortaleza with exacerbation of allergic symptoms during cashew flowering period, the aim of this study was to produce a protein extract from cashew tree pollen and identify the allergens not yet studied. Twelve patients living in Fortaleza, Northeast of country, were selected based on history of persistent allergic rhinitis and aggravation of symptoms at the time of cashew tree flowering. Another group living in the same region with allergic rhinitis without clinical relation with the flowering season was selected. Also 5 non-atopic subjects exposed to cashew tree were selected as a control group. The serum of these patients was tested for 1D and 2D Western Blotting (WB) and selected proteins were submitted to mass spectrometry for identification. Epitopes were predicted by in silico search comparing detected sequences against epitope databases. It was possible to identify some homologs of allergens from other pollens such as isoflavone reductase (Bet v 6), beta-1,3-glucanase (Ole e 9), heat shock protein 70kDa (Cor a 10), besides other proteins that might represent novel allergens, such aminociclase, glutamina sintetase, phosphoglucomutase, alpha-1,4-glucan-protein-synthase, elongation factor 2 and biotin carboxylase among others. The epitope prediction revealed the possibility of cross-reactivity with other known pollen allergens such as Phl p 4, Mal d 1 and other aeroallergens also appeared. This is the first description of cashew pollen allergy showing specific IgE reactivity of patients\' sera. The immunological and structural characterization of new allergens, besides aiding the diagnosis and treatment of non-described allergies, offers tools for predicting epitopes and producing hypoallergenic molecules in this era of precision medicine

A common characteristic of all chemical allergens, both respiratory and skin allergens, is the ability to form stable associations with proteins; the resulting hapten-protein complex being sufficient to provoke an immune response. There is evidence to suggest that selective binding of chemicals with proteins or peptides may impact on the quality of immune response that will develop. In the investigations described here, we have taken a reductionist approach to protein reactivity and evaluated the binding characteristics of 20 of the most commonly reported chemical respiratory allergens towards defined peptides with a single reactive amino acid of interest. The hypothesis is that it is possible to identify and characterize different forms of chemical allergens as a function of preferential peptide binding.Utilizing the standardized reaction conditions of a direct peptide reactivity assay (DPRA), the reactivity of respiratory allergens for cysteine and lysine peptides was evaluated. Activity in the DPRA is reported as the percent depletion of peptide following 24 h incubation. An important and intriguing observation was that, when compared with skin sensitizers, chemical respiratory allergens exhibited a preferential reactivity for lysine. This preference was characterized quantitatively as a ratio of the mean depletion of lysine compared with cysteine (Lys:Cys ratio). The Lys:Cys ratio was observed to be robust and reproducible over time.A limitation of many in chemico methods for hazard identification is the lack of a metabolic component that allows for the identification of pro-haptens. In order to address this limitation, reported here is the use of the peroxidase peptide reactivity assay (PPRA), which utilizes a horseradish peroxidase/hydrogen peroxide (HRP/P) enzymatic system as a proxy for oxidative metabolism. Additionally, reactivity in the PPRA is characterized after a 24 h reaction time utilizing a concentration-response model (thus, permitting consideration of dose-response relationships defined as an EC15 value). Unexpectedly, the preferences for lysine observed with chemical respiratory allergens in the DPRA were lost or blunted in PPRA. The EC15 values demonstrated that relative reactivity between chemical respiratory allergens varied by up to 4 orders of magnitude. The identification of quantitative differences in reactivity could prove useful as a guide to evaluate potency in the future, should reliable metrics become available.To characterize the selectivity of binding by chemical respiratory allergens, the DPRA was modified to allow for the evaluation of reactivity to histidine, tyrosine and arginine. Confirming our previous observations, each of the respiratory sensitizers was observed to react to both lysine and cysteine, with in most instances, a preference for the former. Reactive promiscuity was a function of the other peptides with histidine being the most reactive followed by arginine and tyrosine. To model more complex reactive conditions, a novel modification was made to the DPRA to allow competition for lysine and cysteine to be assessed in a single reaction mixture. The results of these competitive reactivity experiments identified a range of binding patterns to lysine and cysteine that in some cases resulted in different binding being expressed.At present, there are no methods available to reliably identify potential chemical respiratory allergens. The work presented here has demonstrated that respiratory allergens can be identified as potential sensitizers based on their ability to react with lysine and cysteine. More importantly, the balance of reactivity to these two peptides can provide a means of discriminating between respiratory and skin sensitizers.

Germination of mung bean, Phaseolus mungo L. , and wheat, Triticum aestivum L., seeds was used for bioassay to demonstrate the toxic effects of selected pesticides--menazon, disulfoton, and GS-14254-- and allergens-- alantolactone and usnic acid. The ability of gibberellic acid to counteract the toxic effects of these chemicals on germination and seedling growth was studied. Chemical composition of the treated and untreated seeds was made with special attention to starch and protein degradation. Effect of these toxicants on the synthesis of amylase, ATPase, and protease enzymes during germination was studied since these enzymes are synthesized de novo during germination. To ascertain their effect on protein synthesis in storage tissue of the germinating seeds, uptake and incorporation of 14C-L-leucine into protein was studied in potato tuber slices and germinating mung beans. Correlation of biochemical data and histochemical changes in the treated and untreated seeds of mung bean was obtained with menazon and usnic acid. Furthermore, ultrastructural changes were studied in order to relate functional and structural changes in the seeds .in conjunction with phytotoxic actions of these chemicals. Among the insecticides, menazon (250 ppm) was found to be more toxic to both species than was disulfoton. GS-14254 (100 ppm) also was equally inhibitory to seed germination and seedling growth of mung bean and wheat seeds. When a solution of the herbicide GS-14254 (100 ppm) was added to either of the insecticides at their maximum concentrations the inhibitory effect of the combined pesticides on seed germination and seedling growth was more pronounced, especially with wheat. Usnic acid (50 to 250 mg/1) and alantolactone (100 mg/1) significantly inhibited germination and root and shoot growth in both mung bean and wheat seeds. These two compounds appeared to be more phytotoxic than the pesticides. Gibberellic acid partially counteracted the inhibitory effects of the pesticides and allergens, thus these chemicals showed no antiauxin activity. Before any growth is observed there is a marked increase in respiration during germination that releases energy from food materials already present in usable form in the cells. At their maximum concentrations, menazon, disulfoton, GS-14254, alantolactone, and usnic acid significantly blocked the respiration of the germinating seeds at the end of 72 h after treatment. In all cases except alantolactone respiration of wheat seeds was considerably more affected than that of the mung beans. Compared to control seeds, pesticide chemicals as well as allergenic compounds caused significant reduction in the amounts of soluble reducing sugars and free amino acids after 72 h germination period. Similarly, starch degradation was less in the treated seeds. Among the species of seeds, considerably less amounts of reducing sugars and amino acids were formed in the pesticide-treated wheat seeds than in the mung beans as compared to their respective controls. Such differences in the inhibitory effects were not observed in seeds treated with allergenic compounds. The development of amylase and ATPase activity in the seeds treated with maximum concentrations of pesticides tended to be lower than that in the control seeds. In case of menazon, inhibition of amylase activity was more pronounced than that of disulfoton or GS-14254. Proteoiytic activity in control and disulfoton- and menazon-treated seeds was not significantly different during germination period, but in case of GS-14254, it was considerably lower. Usnic acid at highest concentration tested completely inhibited the development of amylase activity in mung beans whereas it was significantly lower in seeds treated with the maximum concentration of alantolactone. The inhibition of amylase activity in wheat seeds treated with these compounds was more or less similar. ATPase inhibition in seeds treated with usnic acid was more severe than that in alantolactone treated seeds. However, proteolytic activity in control and treated seeds showed almost the same trend during the germination period. The activity per se of amylase isolated from mung bean and wheat seeds germinated for 3 days was not significantly inhibited by the presence of the pesticides or allergens in the reaction mixture indicating that these chemicals do not inhibit already synthesized amylase enzyme. Observations with potato tissue and germinating mung beans indicated that both total uptake and incorporation of 14c-L-leucine into protein were significantly inhibited by menazon, disulfoton, GS-14254, and alantolactone. On the other hand, the uptake in germinating mung bean treated with usnic acid was not affected although both uptake and incorporation were inhibited in potato tissue. Menazon and usnic acid were then selected as the representative chemicals for pesticides and allergens, respectively, and their toxic effects were studied histochemically in 3-day germinating mung beans. It was observed that total nucleic acid content and RNA content in seeds treated with these chemicals were considerably less than that in the control seeds. Similarly, treated seeds showed more starch grains and protein bodies indicating less metabolic activity in these seeds. At the ultrastructural level, menazon- or usnic acid-treated mung bean cotyledons at day 3 of germination contained no vacuoles but many undigested protein bodies were observed. In contrast, fully developed mitochondria, endoplasmic reticulum with ribosomes, and vacuoles were seen in control cells indicating protein (enzyme) synthesis and digestion of the food reserves.

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