Introdução: A Agamaglobulinemia, a Imunodeficiência Comum Variável (IDCV) e a Síndrome Hiper IgM (SHIGM) são imunodeficiência primárias predominantemente humorais que se beneficiam da reposição de imunoglobulina humana, com redução da morbimortalidade. No entanto, apesar da reposição adequada de imunoglobulina, complicações pulmonares podem ocorrer, influenciando o prognóstico destes pacientes. Objetivo: O objetivo do estudo foi descrever as alterações morfológicas e funcionais pulmonares em pacientes com hipogamaglobulinemia primária em tratamento com reposição de imunoglobulina humana. Métodos: Foram avaliados 30 pacientes (agamaglobulinemia n=14; IDCV n=9; SHIGM n=7) que receberam imunoglobulina e antibioticoterapia profilática regularmente. A avaliação utilizou dados dos prontuários sobre o início e a evolução da doença, bem como dados de espirometria e tomografia computadorizada de tórax. O escore de Bhalla foi aplicado à tomografia mais recente de cada um dos pacientes para correlacionar as alterações tomográficas pulmonares com os dados clínicos, resultados das espirometrias e ocorrência de processos infecciosos sino-pulmonares após o início da reposição de imunoglobulina. Para as análises estatísticas utilizou-se o programa SPSS 13.0, e valores de p<0,05 foram considerados estatisticamente significantes. As variáveis nominais foram comparadas através do teste de Fisher e as contínuas, através de testes não paramétricos (Mann-Whitney, Kruskal- Wallis e Wilcoxon). Para as correlações do escore de Bhalla com as demais variáveis foi utilizado o coeficiente de Spearman. Resultados: Houve diminuição na frequência de pneumonias (p<0,001) e aumento na frequência de sinusites (p<0,001) após o início da reposição de imunoglobulina. Distúrbios ventilatórios foram evidenciados em 14 dos 23 pacientes que puderam realizaram espirometria (7 obstrutivos, 5 restritivos e 2 inconclusivos). Pacientes com bronquiectasias ao diagnóstico e aqueles à primeira avaliação tomográfica apresentaram mediana de idade mais elevada ao diagnóstico (p=0,015 e p=0,001, respectivamente) e tempo mais prolongado entre o início dos sintomas e o diagnóstico que aqueles sem bronquiectasias (p=0,010 e p=0,001, respectivamente). Sete pacientes desenvolveram bronquiectasias durante o tratamento. Pacientes com bronquiectasias à avaliação final apresentaram maior frequência de sinusites antes do início da reposição de imunoglobulina que aqueles sem bronquiectasias (p=0,010). Houve correlação estatisticamente significante do escore de Bhalla com VEF1 pré e pós-broncodilatador (r= -0,778 e r= -0,837, respectivamente), CVF (r= -0,773), FEF25-75% (r= -0,571) e com a frequência de pneumonias após o início do tratamento (r= 0,561). Conclusões: O tratamento com reposição regular de imunoglobulina e antibioticoterapia profilática reduziu a frequência e gravidade das infecções pulmonares, porém não evitou a ocorrência de sinusites, o aparecimento de bronquiectasias nem de outras alterações morfológicas e funcionais pulmonares / Introduction: Agammaglobulinemia, Common Variable Immunodeficiency (CVID) and Hyper IgM Syndrome (HIGM) are predominantly antibody deficiencies that benefit from immunoglobulin replacement therapy, with reduction of their morbidity and mortality. Despite regular immunoglobulin replacement, pulmonary complications may occur in those patients, affecting their prognosis. Objective: The aim of this study was to describe tomographic and functional pulmonary abnormalities in patients with primary hypogammaglobulinemia receiving immunoglobulin replacement therapy. Methods: Thirty patients (agammaglobulinemia n=14, CVID n=9, HIGM n=7) receiving antimicrobial prophylaxis and regular immunoglobulin infusions were evaluated. Clinical records were reviewed to obtain data concerning the onset and evolution of the disease and the results of spirometry and computed tomography of the chest. Bhalla score was applied to the most recent tomography of each patient to correlate tomographic pulmonary abnormalities with clinical data, spirometry results and the occurrence of sinusal and pulmonary infections after the onset of the immunoglobulin replacement. Statistical analysis was performed using the software SPSS 13.0 and p values < 0.05 were interpreted as statistically significant. Nominal variables were tested using Fisher´s exact test and continuous variables were tested using non-parametric tests (Mann-Whitney, Kruskal-Wallis e Wilcoxon). Spearman coefficient was used to correlate Bhalla score with the other variables. Results: The frequency of pneumonias decreased (p<0.001) and the frequency of sinusitis increased (p<0.001) after the onset of immunoglobulin replacement. Pulmonary function was abnormal in 14 of 23 patients (7 obstructive, 5 restrictive, 2 inconclusive). Patients with bronchiectasis at diagnosis and those with bronchiectasis at the first tomographic evaluation presented higher median age at diagnosis (p=0.015 and p=0.001, respectively) and longer duration between the onset of symptoms and diagnosis than those without bronchiectasis (p=0.010 e p=0.001, respectively). Seven patients developed bronchiectasis during treatment. Patients with bronchiectasis at the last tomographic evaluation presented a higher frequency of sinusitis before therapy onset than those without bronchiectasis (p=0.001). Statistically significant correlation was found between Bhalla score and pre and post bronchodilator FEV1 (r= -0.778 and r= -0.837, respectively), FVC (r= -0.773) and FEF25-75% (r= -0.571) and between Bhalla score and the frequency of pneumonias after the onset of immunoglobulin replacement therapy (r=0.561). Conclusions: Immunoglobulin replacement therapy and antimicrobial prophylaxis reduced the frequency and severity of pulmonary infections but did not prevent the occurrence of sinusitis, the development of bronchiectasis or other morphological and functional pulmonary abnormalities

Orientador: João Martins Pizauro Junior / Resumo: O desenvolvimento dos anfíbios é caracterizado por uma série de eventos que possibilitam a esses animais a ocupação dos ambientes aquático e terrestre. Nesse sentido, a supressão do sistema imunológico é necessária para que esses eventos ocorram de forma ordenada, evitando então, respostas autoimunes. Por outro lado essa imunossupressão durante a metamorfose pode tornar os animas mais susceptíveis a algumas doenças, dentre elas, a causada pelo fungo Batrachochytrium dendrobatidis (Bd), que é considerado o principal patógeno de anfíbios da atualidade. Nesse contexto, a produção de anticorpos da classe IgY anti-Bd, extraídos de gemas de ovos de galinhas poedeiras infectadas com o referido fungo, pode agregar valor ao ovo, auxiliar no combate a quitridiomicose, diminuição da perda da produção e dispersão do patógeno por criadouros, diminuindo também o uso de antibióticos no tratamento e profilaxia da quitridiomicose, aumentando o nível de segurança da carne e valorizando o produto final. No presente estudo a imunização das aves com o antígeno desativado fez com que o sistema humoral das mesmas respondesse a indução contra Bd viabilizando a produção e purificação de imunoglobulinas da classe IgY específicas e de alta avidez contra o fungo. A transferência passiva das imunoglobulinas purificadas das gemas apresentou resultados promissores, onde os girinos de rã-touro (Lithobates catesbeianus) tratados não apresentaram danos deletérios quanto à sanidade animal dos mesmos, mantendo ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The development of amphibians is characterized by a series of events that enable these animals to occupy aquatic and terrestrial environments. In this sense, suppression of the immune system is necessary for these events to occur in an orderly manner, avoiding autoimmune responses. On the other hand, this immunosuppression during metamorphosis may make the animals more susceptible to some diseases, among them, the one caused by the fungus Batrachochytrium dendrobatidis (Bd), which is considered the main amphibian pathogen today. In this context, the production of anti-Bd IgY class antibodies, extracted from egg yolks of laying hens infected with this fungus, may add value to the egg, assist in the fight against chytridiomycosis, decrease production loss and pathogen dispersion by breeding facilities, reduce the use of antibiotics in the treatment and prophylaxis of chytridiomycosis, increasing the safety level of the meat and valuing the final product. In the present study, the immunization of hens with the deactivated antigen caused their humoral system to respond to induction against Bd, enabling the production and purification of specific and highly avid fungal IgY class immunoglobulins. Passive transfer of purified immunoglobulins from yolk showed promising results, where treated bullfrog tadpoles (Lithobates catesbeianus) showed no deleterious damage to animal health, keeping the morphology and counting of blood cells within normal range. These results enable the develop... (Complete abstract click electronic access below) / Doutor

Includes bibliographies. / [xvii], 173, [38] leaves, [18] leaves of plates : / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis presents the development of an assay for reverse transcription-polymerase chain reaction (RT-PCR) to be applied to the immunomagnetic isolation of carcinoma cells, as a possible means of detecting small numbers of breast cancer cells in a haemopoietic environment. The messenger RNA expression of two different genes, CD44 and the cytokeratin K19, is assessed for suitability as tumour markers for the Immunobead RT-PCR method, and clinical results using K19 are presented. / Thesis (M.D.)--University of Adelaide, Dept. of Medicine, 1977?

Thesis (Ph. D.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on December 12, 2007) Vita. Includes bibliographical references.

El limfoma de cèl•lules del mantell (LCM) és una neoplàsia heterogènia en relació a les característiques clíniques, biològiques i moleculars que presenten els pacients. La morfologia, l’immunofenotip i el genotip no són suficients per explicar aquesta gran diversitat. Encara que es considera una neoplàsia agressiva alguns pacients segueixen un curs clínic indolent sense la necessitat de tractament en el moment del diagnòstic i presenten una supervivència llarga. Els resultats d’aquesta tesi demostren que els LCM amb elevada càrrega mutacional en els gens de les immunoglobulines (IGHV)(<97% d’identitat amb la línia germinal) utilitzen preferencialment certes famílies de gens IGHV, suggerint la presència de processos de selecció antigènica com a mecanisme de patogènesi d’aquesta neoplàsia i donant importància a la caracterització del receptor de les cèl•lules B (BCR). Els LCM amb gens IGHV mutats (M-LCM) presenten una menor complexitat genòmica, són preferencialment negatius per a l’expressió del factor de transcripció SOX11 i presenten afectació ganglionar amb més freqüència que els casos sense mutacions (U-LCM). Per altra banda, el perfil d’expressió gènica entre casos amb i sense mutacions en els gens IGHV, també mostra expressió diferencial de programes cel•lulars relacionats amb poblacions de cèl•lules B normals incidint amb la ontogènia d’ambdós grups. El perfil d’expressió de microRNAs també permet diferenciar entre dos grups de pacients amb diferent supervivència, reforçant la idea de la presència de dos subgrups que pertanyen a una mateixa entitat. La identificació i la correcte identificació d’aquests pacients de limfoma de cèl•lules del mantell són rellevants per al posterior maneig i tractament dels mateixos. / Mantle cell lymphoma is a malignant and heterogeneous disease regarding the clinical, biological and molecular characteristics presented by patients, all of them with the translocation t(11;14) and CCND1 overexpression. Although is considered an aggressive neoplasm some patients follow and indolent clinical course without treatment at the time of diagnosis and long survival. The results of this studies demonstrate that LCM with high mutational load in the immunoglobulins genes (IGHV) (<97% of identity with the germ line) use preferentially specific IGHV gene families, suggesting the presence of antigenic selection process as a mechanism of pathogenesis of this neoplasm and giving importance to the characterization of the B cell receptor. The LCM cases with IGHV mutated genes (M-LCM) present less genomic complexity are preferentially negative for the expression of the transcriptor factor SOX11 and also present lymphadenopathies more frequently compared with the IGHV unmutated cases (U-LCM). Furthermore, the gene expression profile between cases with and without IGHV mutations also shows differential expression of cellular programs related to normal B cell populations maybe related with the ontogeny for both groups. The expression profile of microRNAs also differentiates between two groups of patients with different survival and other molecular characteristics, reinforcing the idea of the presence of two subgroups that belong to the same entity. The correct identification and classification of these patients are relevant for subsequent handling and treatment of the patients.

Production of xylanase and itaconic acid (IA) from Aspergillus terreus NRRL 1960 from agricultural residues was investigated in this study. Two different media were tested and the medium having itaconic acid inducing capacity was chosen for further studies due to its high xylanase and IA production capacity. The best xylan concentration was found as 2% (w/v). Addition of commercial xylanase to production culture resulted in higher initial simple sugar concentration which increased IA production slightly but decreased xylanase production. Among tested agricultural residues / corn cob, cotton stalk and sunflower stalk, the highest xylanase production was obtained on corn cob. Increasing the corn cob concentration and applying wet heat pretreatment increased the xylanase production level. In a two-step fermentation process, 70000 IU/L xylanase production was achieved in a medium containing 7% wet heat treated corn cob followed by 17 g/L IA production in a medium containing 10% glucose. Molecular weight and isoelectric point of xylanase were found as 19 kDa and pH 9.0, respectively. The enzyme was optimally active at 50&deg / C and pH 6.5-7.0. Kinetic experiments at 50&deg / C and pH 7.0 resulted in apparent Km and Vmax values of 2.5&plusmn / 0.05 mg xylan/mL and 50.2&plusmn / 0.4 IU/&micro / g protein, respectively. The major products of birchwood xylan hydrolysis were determined by thin layer chromatography as xylobiose and xylotriose. These findings indicate that the enzyme could be advantageous for use in different industrial applications due to its low molecular weight and its potential use for xylooligosaccharide production.

Fast and accurate detection of pathogens such as bacteria, their toxins and viruses at low concentrations is very important. The conventional techniques are time consuming where expensive equipment is required with a consumption of excess amount of blood from patients. Recently, immunosensors are used for the detection of pathogens because they are miniature, sensitive, biocompatible and require low power. According to the Centers for Disease Control and Prevention (CDCP), 76 million people become ill due to food poisoning and 5,000 of them die each year in United States. In addition, SEB causing food poisoning has listed as a bioterrorism agent by CDCP. Thus, accurate and selective detection in short time is very important for SEB detection. Candida albicans (C. albicans) is a yeast-like fungus and causes anxiety, insomnia, constipation, hiatal hernia, panic attacks, denture-induced stomatitis, angular cheilitis, gingivitis and prosthetic implant infections. In addition, it can cause death if the immune system of patient is under failure due to cancer, chemotherapy and AIDS. In this study, a new procedure was developed. A simple and highly selective homogeneous sandwich immunoassay was obtained for ultrasensitive detection of Staphylococcal Enterotoxin B (SEB) using Atomic Force Microscopy (AFM) and Surface Enhanced Raman Scattering (SERS) probe. In the developed procedure, thiolated antibodies were produced and SEB was immobilized on the biosensor surface using these antibodies. In addition, theory of SEB adsorption on a gold surface was studied and the reaction rate constant between SEB and its toxin was calculated. Moreover, C. albicans was detected using the developed procedure by a microscope. Thus, it is proved that, the developed procedure can be used for detection of different pathogens. Furthermore, nonspecific interaction between SEB antibody and BSA was determined in this study. Also, the developed procedure and a procedure found from literature were compared. In the procedure used in the literature (second procedure), self-assembled monolayer (SAM) was formed and antibodies were immobilized on SAM. After formation of sandwich structure, the roughness of gold surface and the minimum concentration of SEB detected were determined by AFM and SERS, respectively.

The protozoan parasite Trypanosoma congolense is one of the aetiological agents of African animal trypanosomiasis that is transmitted by the tsetse fly. The parasite causes nagana in animals and affects livestock throughout sub-Saharan Africa. The toxicity of available drugs and the emergence of drug resistant parasites have affected the treatment of trypanosomiasis. Control of the disease has also been difficult due to ineffective vector control and the potential of trypanosomes to express hundreds of antigenetically distinct proteins on their surface. Vaccination against trypanosomiasis has been thought to be a possible control method. Since a vaccine based on variable surface proteins of the parasite is unlikely, research has been directed towards the identification of invariant pathogenic factors of the parasite as potential targets for therapy. Congopain, the major cysteine protease of T. congolense has been implicated in the pathology of the disease. Antibodies against congopain are known to contribute to the mechanisms of natural resistance to trypanosomiasis known as trypanotolerance by neutralising the pathogenic effects of the enzyme. Oligopeptidase B (OpdB), a trypanosomal serine protease has also been associated as a pathogenic factor of the disease. It is released into the host’s circulation by dead or dying parasites and retains its catalytic activity since it is insensitive to host serum inhibitors. In the present study, the catalytic domain of congopain (C2) and the use of alpha-2-macroglobulin (α2M) as an adjuvant were investigated for their potential use in an anti-disease vaccine. α2-Macroglobulin has been used to varying degrees to target different antigens to cells of the immune system and enhance their immunogenicity. A previous study showed that antibodies raised in rabbits against C2 complexed to α2M gave a higher percentage inhibition than antibodies made using C2 mixed with Freund’s adjuvant. In the present study, goats were immunised with C2 complexed with α2M to confirm the enhanced immunogenicity of C2 and the production of anti-C2 antibodies with superior inhibitory properties. Following immunisation, goats were challenged with T. congolense (strain IL 1180) and showed sustained antibody production during the two month infection period. Goat antibodies made using C2 in complex with α2M inhibited the hydrolysis of hide powder azure by C2 by 96%. Maximum inhibition of the hydrolysis of azocasein was observed to be 63% and hydrolysis of Z-Phe-Arg-AMC by C2 was inhibited by 73%. In order to determine the vaccine potential of OpdB, protein was recombinantly expressed as a glutathione-S-transferase fusion protein in the pGEX expression system and purified by glutathione agarose affinity chromatography and molecular exclusion chromatography. Since a small yield of protein necessitated several rounds of expression and extensive purification, OpdB was subsequently expressed as a His-tagged fusion protein in the pET bacterial expression system. Recombinant protein was easily purified using nickel chelate affinity chromatography. Purified OpdB was used with alum for the immmunisation of mice to produce antibodies capable of inhibiting enzyme activity. Following immunisation, mice were challenged with T. congolense (strain IL 1180) and also showed sustained antibody production following two months infection. Since all mice died, the administration of OpdB conferred no protection; however, anti-OpdB mouse antibodies inhibited 86% of OpdB activity against the substrate Z-Arg-Arg-AMC. In addition immunised mice were observed to survive 40% longer than control mice as they had previously been immunised with OpdB and were able to mount a rapid immune response against this pathogenic factor during infection. In general it could be concluded that immunisation of goats with C2 in complex with α2M produced antibodies with superior inhibitory properties. The immunisation of mice with OpdB and alum also produced inhibitory antibodies and previous administration of OpdB enabled mice to mount a rapid immune response against OpdB during infection. Antibody mediated enzyme inhibition demonstrates the potential use of C2 and OpdB as vaccines that may contribute to the development of an effective anti-disease vaccine. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Diss. (sammanfattning) Uppsala : Univ., 2002. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.