Made available in DSpace on 2014-12-17T14:13:32Z (GMT). No. of bitstreams: 1 MarcioLTA_Tese.pdf: 1453040 bytes, checksum: 7c0ed7c44a56eb54d2170012eedbfa04 (MD5) Previous issue date: 2009-11-27 / The imprecision of the frontier that separates those cognitive deficits inherent to the human physiological aging process from those which represent the early signs of nervous system degenerative pathologies ,very prevalent among the elderly, has brought attention to the need of studies aiming to establish clinical and/or laboratorial criteria to allow this differentiation. Elderly people living in poor and developing countries are frequently exposed to precarious socioeconomic conditions which facilitate the development of an array of pathologies which have metabolic and nutritional dysfunctions as the established or proposed etiological agents. The levels of certain micronutrients, such as the vitamins B12 and B9 (folic acid), and of some intermediary metabolites, such as homocysteine are being thought of as etiological factors and/or as biological markers of a group of alterations which affect the normal functioning of the nervous system with important reflexes upon cognitive performance. This study aims to investigate the influence of homocysteine, B12 vitamin and folic acid levels on the cognitive performance of the low income elderly population. This transversal study took place in Natal, Rio Grande do Norte State, Brazil, and involved 205 dwelling elderly people, users of the Programa de Sa?de da Fam?lia, a public healthcare program, maintained by the city s health authorities. A multidimensional questionnaire was used to assess the socio-demographic aspects and the overall health and nutrition conditions. The cognitive performance was measured by the use of the Portuguese version of the Mini Mental State Exam (MMSE). The serum levels of homocysteine, B12 vitamin and folic acid were determined by chemiluminescence. The association between the socio-demographic and serum levels of Hcy, B12 vitamin and folic acid was determined by multiple linear regression. Serum levels higher than 13.5 &#956;mol/l, indicative of hyperhomocysteinemia (HHcy), were found on 25.4% of the sample, being more prevalent in men (p<0.05). Deficitary levels of folic acid (<5ng/mol) and of B12 vitamin (<193 pg/ml) were found on 3.9% and 10.2% of the sample respectively. A negative correlation was found between cognitive performance with both age and HHcy and a positive correlation was found between cognitive performance and schooling. The isolated HHcy R2 values were an explanation to only 4% of the variance of the MMSE scores. However, when associated with schooling and age, this model explains about 25% of this association / A imprecis?o da fronteira que separa os d?ficits pr?prios do processo de envelhecimento fisiol?gico humano daqueles que representam os sinais precoces das patologias degenerativas de grande preval?ncia entre idosos, tem chamado a aten??o para a necessidade da produ??o de estudos voltados para o estabelecimento crit?rios cl?nicos e/ou laboratoriais que permitam essa diferencia??o. Idosos de popula??es de pa?ses pobres e/ou em desenvolvimento s?o freq?entemente expostos a condi??es socioecon?micas prec?rias prop?cias ao desenvolvimento de um conjunto de patologias, disfun??es metab?licas e nutricionais. Os n?veis de determinados micronutrientes e de alguns metab?litos intermedi?rios v?m sendo vistos como fatores etiol?gicos e como marcadores biol?gicos de um conjunto de altera??es que afetam a fun??o normal do sistema nervoso com reflexos importantes sobre o desempenho cognitivo. N?veis elevados de homociste?na (Hcy) e d?ficits nutricionais e /ou metab?licos da vitamina B12 e B9 (?cido f?lico) t?m sido apontados como preditores e/ou como fatores etiol?gicos de altera??es cognitivas. O objetivo desse estudo foi avaliar a influ?ncia dos n?veis de homociste?na, Vitamina B12 e ?cido f?lico no desempenho cognitivo de idosos de baixa renda. Este estudo transversal desenvolvido em Natal, Rio Grande do Norte, Brasil, incluiu 205 idosos n?o institucionalizados atendidos pelo Programa de Sa?de da Fam?lia da Secretaria Municipal de Sa?de do munic?pio. Um question?rio multidimensional foi utilizado para avaliar os aspectos sociodemogr?ficos e as condi??es gerais de sa?de e nutri??o. O desempenho cognitivo foi aferido utilizando-se a vers?o em portugu?s do Mini-Exame do Estado Mental (MEEM). Os n?veis s?ricos de homociste?na, Vitamina B12 e ?cido f?lico foram determinados por quimioluminesc?ncia. A associa??o entre as vari?veis sociodemogr?ficas e os n?veis s?ricos de Hcy Vitamina B12 e ?cido f?lico foi determinada atrav?s de regress?o linear m?ltipla. Valores s?ricos acima de 13,5 &#956;mol/l indicativos de hiperhomocisteinemia (HHcy) foram encontrados em 25,4% da amostra sendo mais prevalente em homens (p<0,05). N?veis deficit?rios de ?cido f?lico (<5ng/ml) e de Vitamina B12 (<193 pg/ml) foram encontrados em 3,9% e 10,2% dos indiv?duos respectivamente. O desempenho cognitivo mostrou uma correla??o negativa com a idade e a HHcy e positiva com a escolaridade. Os valores R2 da HHcy isolada explicaram apenas 4% da vari?ncia da pontua??o do MEEM. Contudo, quando associada escolaridade e idade, este modelo explica aproximadamente 25% desta associa??o.

Made available in DSpace on 2014-08-20T13:32:50Z (GMT). No. of bitstreams: 1 tese_vinicius_coitinho_tabeleao.pdf: 864855 bytes, checksum: 84b7f4f2dc9f4bc7025094cdcd06be5f (MD5) Previous issue date: 2014-02-28 / Dairy cows are expected to perform maximum of their productive capacity, especially during the transition period (three weeks before and up to three weeks after calving). This period is characterized by intense changes in metabolism which cause a negative energy balance. This fact generates a period of typical weight loss, which has been correlated with insulin resistance, once the available glucose is destined for lactose synthesis, which in turn is responsible, in part, for milk production levels. Moreover, during lactation there are several managements that impairs animals health. If not executed properly, can cause trauma and predisposition for infections, especially of the mammary gland, because this gland is heavily manipulated and has contact with potentially infectious agents. From a entrance doorway, these agents can infect the gland and cause mastitis, which may be clinical (with clinical signs) or subclinical (without clinical signs). In situations of metabolic disorders or in cases of mastitis, the milk production may be affected. Therefore, there is a constant search for technologies that can minimize these disorders and assist the production systems. Our objective was to investigate the effects of combining butaphosphan and cyanocobalamin on the glycemic metabolism in healthy dairy cows in the postpartum period and their effects on the mammary gland recovery in dairy cows after clinical mastitis treatment. Thus, our study was stratified into two assumptions: 1) the use of the association of butaphosphan and cyanocobalamin enhances peripheral insulin resistance in dairy cows, providing greater amounts of glucose for milk production, 2) the supplementation with butaphosphan and cyanocobalamin facilitates mammary gland recovery in dairy cows affected by mastitis. In experiment 1, animals receiving repeated doses of butaphosphan and cyanocobalamin had greater weight loss and increased (P<0.05) of the activities of the enzymes aspartate aminotransferase and creatine kinase. Moreover, at end of treatment had an increase (P<0.05) in glucose concentration in treated animals when compared to the beginning of study. In experiment 2, the comparative analyzes between mild and moderate mastitis did not differ (P>0.05) in any parameters. The animals treated with butaphosphan and cyanocobalamin had lower (P<0.05) somatic cell count after the third application. However, the concentrations of glucose, albumin, TPP, NEFA and the chemical and physical components of milk did not differ (P>0.05) among groups. In concusion, the combination of butaphosphan and cyanocobalamin shows promising results as a source of nutrient that allows intensify physiological reactions mainly dependent phosphorylation. / As vacas leiteiras são desafiadas a desempenhar o máximo da sua capacidade produtiva, principalmente, durante o período de transição (três semanas anteriores e três semanas posteriores ao parto). Este período é caracterizado por intensas mudanças no metabolismo que causam um balanço energético negativo. Este fato gera uma perda de peso característica do período, que tem sido correlacionada com a resistência periférica à insulina, haja vista, que a glicose disponível esta sendo destinada para síntese de lactose, que por sua vez é responsável, parcialmente, pelo volume de leite produzido. Além disso, durante a lactação há vários manejos que podem oferecer risco à saúde dos animais, caso não sejam executados adequadamente, causando traumas e predispondo o animal a infecções, especialmente, da glândula mamária, já que ela é intensamente manipulada e tem contato com vários agentes potencialmente infecciosos. A partir de uma porta de entrada estes agentes podem infectar a glândula causando a mastite, que pode se apresentar na forma clínica (com presença de sinais clínicos), ou subclínica (sem sinais clínicos). Nas situações de desequilíbrio metabólico, ou em casos de mastite, a produção de leite pode ser prejudicada. Diante disso, há uma busca constante por tecnologias que possam minimizar estes transtornos e auxiliar os sistema de produção. Nosso objetivo foi verificar o efeito da associação de butafosfan e cianocobalamina sobre o metabolismo glicêmico de vacas leiteiras saudáveis no período pós-parto, além dos seus efeitos sobre a recuperação da glândula mamária em vacas leiteiras após o tratamento de mastite clínica. Para tanto, nosso estudo foi estratificado em dois trabalhos com as seguintes hipóteses: 1) a utilização da associação de butafosfan e cianocobalamina pode intensificar a resistência à insulina periférica, disponibilizando maior quantidade de glicose para a produção de leite em vacas leiteiras; 2) a suplementação da associação de butafosfan e cianocobalamina pode auxiliar na recuperação da glândula mamária de vacas leiteiras, após o tratamento da mastite clínica. No experimento 1, os animais que receberam sucessivas doses de butafosfan e cianocobalamina tiveram maior perda de peso e aumento (P<0,05) das atividades das enzimas aspartato amino transferase e creatinoquinase. Além disso, ao final do tratamento houve aumento (P<0,05) dos níveis glicêmicos nos animais tratados, quando comparados ao inicio. No experimento 2, as análises comparativas entre as mastites, leve e moderada, não diferiram em nenhum dos parâmetros (P>0,05). Os animais tratados com butafosfan e cianocobalamina apresentaram menor (P<0,05) contagem de células somáticas após a terceira aplicação. Entretanto, os níveis de glicose, albumina, PPT, AGNES e os componentes químicos e físicos do leite não diferiram (P>0,05) entre os grupos. Nessas condições de estudo, podemos concluir que a associação de butafosfan e cianocobalamina apresenta resultados promissores como fonte de nutriente que permite intensificar as reações fisiológicas, principalmente, as dependentes de fosforilação.

Thesis (MSc) -- Stellenbosch University, 1990. / ENGLISH ABSTRACT: The main aim of this workpiece was to establish the physiological parameters against which a vitamin Bu deficiency could be measured. A comparison between the hematological values of healthy patients and those suffering from pernicious anemia due to vitamin Bu deficiency was done. A specific case of pernicious anemia was used in the comparison of abnormal values to the values of normal healthy patients. The comparison consisted of blood analyses with the help of specified instruments, photomicrographs of bone marrow and blood smears and statistical data. A Coulter Counter Model ZF was used for the hematological analyses of blood, a radio-isotope assay for serum vitamin B u was done and photomicrographs were taken with a NIKON Microflex camera with photomicrographic attachments. The importance of vitamin Bu has been shown in this workpiece. With the use of techniques and certain instruments, the effects of a shortage of vitamin Bu has been shown. Analyses of the blood from normal ,healthy patients was compared to that of patients suffering from pernicious anemia. It was demonstrated that pernicious anemia is characterized by a low erythrocyte count, hematocrit (Het), hemoglobin (Hb) and vitamin Bu levels together with a higher mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV). In severe cases of pernicious anemia these levels are extremely high or low as the case may be. Together with these values, the investigation of pernicious anemic blood and bone marrow smears revealed abnormally large erythrocyte precursors and fewer leucocytes than normal. Abnormally shaped cells, called macrocytes, were seen which was due to the disruption in deoxyribonucleic acid (DNA) synthesis caused by the vitamin Bu deficiency. This study produced a set of hematological reference values. The comparative study between healthy and pernicious anemic patients demonstrated a significant drop in serum vitamin B12 values during pernicious anemia. The hematological effect was illustrated by the Coulter Counter blood analysis results and the microscopic examination revealed the presence of megaloblastic erythrocytes, oval erythrocytes, pear-shaped poikilocytes and polymorphonuclear neutropbils with hypersegmented nuclei in blood smears I during vitamin B12 deficiency. This dianoses can be supported by the presence of megaloblasts and metamyelocytes in pernicious anemic bone marrow. / AFRIKAANSE OPSOMMING: Die hoof doel van hierdie werkstuk was om fisiologiese grense te bepaal waarteen 'n vitamien B12 tekort gemeet kan word. 'n Vergelyking tussen die hematologiese waardes van gesonde persone en die van pasiente met pernisieuse anemie wat ontstaan het as gevolg van 'n vitamien B12 tekort was uitgevoer. Die waardes verkry vanaf 'n spesifieke geval van pernisieuse anemie. was vergelyk met waardes vanaf normale gesonde persone. Hierdie vergelyking het bestaan uit bloed analises, fotomikrograwe van bloed en beenmurg smere en statistiese data. Die hematologiese bloed analises was met behulp van 'n Coulter Teller model ZF uitgevoer. 'n Radio-isotoop bepaling vir serum vitamien B12 was gedoen en fotomikrograwe was met 'n NIKON Microflex kamera geneem. Die belang van 'n vitamien B12 tekort was in hierdie werkstuk gedemonstreer. Die effek van hierdie tekort is deur die gebruik van sekere instrumente en tegnieke aangedui en die resultate hiervan is vergelyk tussen gesonde persone en pasiente met 'n vitamien B12 tekort. Hierdie studie het bevestig dat pernisieuse anemie gekenmerk word deur verlaagde eritrosiet, hematokrit (Het), hemoglobien (Hb) en vitamien B12 vlakke tesame met verhoogde gemene korpuskulere hemoglobien (GKH) en gemene korpuskulere volume (GKV) vlakke. Gedurende ernstige gevalle van pernisieuse anemie kan hierdie waardes uitermatig hoog of laag wees. Benewens hierdie waardes het die ondersoek van bloed en beenmurg gedurende vitamien B12 tekort, abnormale groot eritrosiet voorgangers en 'n verminderde hoeveelheid leukosiete getoon. Abnormale sel vorms was ook sigbaar a.g.v. die onderbreking in DNA sintese wat deur 'n vitamien B12 tekort veroorsaak word. Pernisieuse anemie word verkry wanneer daar 'n vitamien B12 en/of folaat tekort in die dieet is of wanneer hierdie vitamiene nie geabsorbeer kan word nie. Die teenwoordigheid van makrosiete, ovaal eritrosiete, peervormige poikilosiete en polimorfonuklere neutrofiele met hipergesegmenteerde keme in bloedsmere dui op 'n megaloblastiese anemie. Hierdie diagnose kan ondersteun word deur die aanwesigheid van megaloblaste en reuse metamielosiete in die beenmurg. Die bepaling van vitamien B12 en folaat vlakke in die bloed kan as addisionele bewysstukke vir 'n volledige diagnose dien. Gedurende hierdie studie is daar 'n stel hematologiese verwysingswaardes vasgestel. Die vergelykende studie tussen gesonde persone en pasiente met pernisieuse anemie het getoon dat daar 'n beduidende verlaging in serum vitamien B12 waardes gedurende pernisieuse anemie is. Die hematologiese effek was ook duidelik waameembaar in die Coulter teller se bloed analiese en mikroskopiese ondersoeke het die · teenwoordigheid van makrosiete, ovaal eritrosiete, peervormige poikilosiete en polimorfenuklere neutrofiele met hipersegmenteerde keme in bloedsmere aangedui. Hierdie diagnose kan ondersteun word deur die aanwesigheid van megaloblaste en reuse metamielosiete in die beenmurg. / This study was financially aided by a bursary from the CSIR.

Based upon experimental evidence from the 1970’s we proposed that a reduced form of hydroxocobalamin should be capable of producing carbon dioxide (CO2) from carbon monoxide (CO) in blood, and that this conversion should be detectable. Using resonance raman spectroscopy we demonstrated that a mixture of hydroxocobalamin and ascorbic acid could create the reduced form of hydroxocobalamin. We used a closed-loop circulation system with a hollow-fiber membrane oxygenator to produce carboxyhemoglobin. Using sensitive gas monitoring equipment to the gas-out port of the oxygenator we analyzed the CO and CO2 concentrations coming from the oxygenator. The mixture of hydroxocobalamin and ascorbic acid caused a 5-fold increase in the CO2 concentration of the gas-out flow, in comparison to baseline and negative controls. These findings offer initial support for the potential use of a mixture of hydroxocobalamin and ascorbic acid as an injectable antidote for carbon monoxide poisoning.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

No description available.

Les folates (vitamine B9) et la vitamine B12 interviennent comme cofacteurs dans le métabolisme des monocarbones qui régule les réactions de transméthylation impliquées dans les mécanismes épigénétiques. Un déficit en folates et/ou B12 réduit la production de méthionine à partir de l'homocystéine, un acide aminé toxique dont l'accumulation a été associée à la survenue de pathologies du système nerveux central aux différents stades de la vie (spina bifida, maladie d'Alzheimer…). Afin d'explorer les mécanismes cellulaires et moléculaires impliqués dans la réponse à la carence en ces micronutriments, nous avons développé deux nouveaux modèles cellulaires. Ainsi, nous avons étudié les effets d'une déficience en folate sur la prolifération, la différenciation et la plasticité neuronale de progéniteurs neuronaux issus de l'hippocampe d'embryons de rat, la lignée H19-7. Le second modèle correspond à un projet innovant visant à obtenir une déplétion cellulaire en B12 par séquestration membranaire. Il a été obtenu par transfection stable de la lignée de neuroblastome murin NIE-115 dans le but d'induire l'expression d'une protéine chimère contenant le transporteur plasmatique de la vitamine B12, la transcobalamine II, et une protéine d'ancrage membranaire. L'ensemble de ces travaux montre que les altérations du métabolisme des monocarbones associées aux carences répriment la neurogenèse et induisent des troubles de la différentiation neuronale. Ceci suggère l'existence de mécanismes précis par lesquels le déficit en folates, en vitamine B12 et/ou l'homocystéine peuvent affecter le fonctionnement du cerveau et sa plasticité. / Folate (vitamin B9) and vitamin B12 act as cofactors in the one-carbon metabolism that regulates transmethylation reactions involved in epigenetic mechanisms. A deficiency in folate and/or B12 decreases the generation of methionine from homocysteine, a toxic amino acid that has been associated with pathologies of the central nervous system at all ages (spina bifida, Alzheimer's disease…). In order to depict the cellular and molecular mechanisms implicated in the response to the deficiency in these micronutrients, two new cell models have been developed. Thus, we have analyzed the effects of folate deficiency on proliferation, differentiation and neuroplasticity of neuronal progenitors obtained from the hippocampus of rat embryos, i.e. the H19-7 cell line. Regarding the second model, we designed an original approach by stable transfection of NIE-115 murine neuroblastoma cells to impose the anchorage of a chimeric B12 binding protein, transcobalaminoleosin (TO) to intracellular membrane in order to produce intracellular sequestration of B12. Taken together, our results have shown that deficiency-associated alterations of the one-carbon metabolism lead to reduced neurogenesis and to dramatic impairment of neuron differentiation. This suggests the existence of specific mechanisms through which vitamin B9/B12 deficiency and/or homocysteine may affect brain functioning and plasticity.

La carence en donneurs de méthyle est assez fréquente dans la période périnatale et au cours du vieillissement. Les donneurs de méthyle alimentaires, l'acide folique et la vitamine B12, influencent la teneur cellulaire en S-adénosylméthionine et S-adénosylhomocystéine et en homocystéine. Le lien entre les donneurs de méthyle et le métabolisme énergétique n'est pas connu, en dépit de leur rôle dans les voies liées à l'épigénétique et la synthèse de molécules méthylées. Nous avons évalué les conséquences d'un régime alimentaire déficient donneur de méthyle, dans le myocarde des ratons au moment du sevrage, soumis à une carence durant la gestation et la lactation. Le régime carencé augmente l'homocystéine plasmatique et S-adénosylhomocystéine myocardique. Les résultats mettent en évidence une cardiomyopathie hypertrophique et un déficit global de l'oxydation des acides gras avec acylcarnitines plasmatiques. Les liens entre le métabolisme des mono-carbones, l'oxydation mitochondriale des acides gras et de cardiomyopathie ont été constatées par des corrélations entre l'homocystéine et les acylcarnitines et le peptide natriurétique de type B. La carence en donneurs de méthyl peut être une condition aggravante de la cardiomyopathie en altérant l'oxydation des acides gras à travers une expression modifiée de PPAR[alpha] et ERR[alpha] et un déséquilibre de l'acétylation / méthylation de PGC1[alpha]. Nous avons montré que l'Hcy et Apo A-I ont été deux facteurs métaboliques déterminants de l'ABI dans une population ambulatoire de volontaires âgés d'une région rurale de la Sicile. L'influence négative de l'Hcy sur Apo A-I et sur le métabolisme des HDL ouvre des nouvelles perspectives sur l'implication de l'Hcy dans la physiopathologie de l'athérothrombose / The deficiency in methyl donors is prevalent in the perinatal period of life and in aging. Dietary methyl donors, folate and vitamin B12, influence the cellular content in Sadenosylmethionine and S-adenosylhomocysteine and increases homocysteine. The link between methyl donors and energy metabolism is not known, despite their role in pathways related to epigenetics and synthesis of methylated molecules. We evaluated the consequences of a diet lacking methyl donors, in myocardium of weaning rats from dams subjected to deficiency during gestation and lactation. The deficient diet increased plasma homocysteine and myocardium Sadenosylhomocysteine. The results evidenced a hypertrophic cardiomyopathy with a global deficit in fatty acid oxidation with increased plasma acylcarnitines. The links between one-carbone metabolism, mitochondrial fatty acid oxidation and cardiomyopathy were ascertained by correlations between hyperhomocysteinemia, short-, medium- and long-chain acylcarnitines, and type-B natriuretic peptide. Methyl donor deficiency may be an aggravating condition of cardiomyopathy by impairing fatty acid oxidation through altered expression of PPAR[alpha] and ERR[alpha] and imbalanced acetylation/methylation of PGC1[alpha]. Finally, in a clinical study we sshowed that the Hcy and Apo A-I were two metabolic factors that influence the « anckle brachial index » in an ambulatory aged Sicilian population. The influence of homocysteine on Apo A-I and HDL metabolism provides new insights on its role on vascular diseases, at a cross-point between atherosclerosis and atherothrombosis

Orientador: Ângelo Berchieri Junior / Banca: Manoel Victor Franco Lemos / Banca: Gerson Nakazato / Resumo: Salmonella enterica sorotipo Typhimurium sintetiza cobalamina (Vitamina B12) apenas sobre condições anaeróbicas. Dois porcento do genoma da S. Typhimurium é dedicado a reações dependentes de vitamina B12 como cofator, sua síntese e absorção. Neste estudo nós preparamos mutantes de Salmonella sorotipos Enteritidis, Gallinarum e Pullorum duplo defectivos na biossintese de cobalamina, cepas ΔcobSΔcbiA. A virulência destes mutantes foi comparada com as respectivas cepas selvagens e, nenhuma deficiência na capacidade de causar doença foi observada para as cepas de S. Enteritidis ΔcobSΔcbiA e S. Pullorum ΔcobSΔcbiA. S. Gallinarum ΔcobSΔcbiA por sua vez, mostrou atenuação total. Posteriormente nós testamos a produção de B12 pelas cepas mutantes e selvagens já descritas, e incluímos neste estudo a cepa de S. Typhimurium ΔcobSΔcbiA, e sua respectiva cepa selvagem. Todas as cepas mutantes não tiveram produção de B12 detectada. As cepas selvagens mostraram produção de vitamina B12 em ambos os ensaios utilizados, com exceção da S. Gallinarum que não apresentou produção de cobalamina in vitro. Como conclusão, a produção de vitamina B12 in vitro diferiu entre os sorotipos de Salmonella testados, a deleção dos genes cbiA e cobS produziu alteração na relação parasita hospedeiro em diferentes níveis entre os sorotipos de Salmonella estudados, sendo que esta foi muito maior entre o sorotipo Gallinarum e as aves. / Abstract: Salmonella enterica serovar Typhimurium only synthesizes cobalamin (Vitamin B12) during anaerobiosis. Two-percent of the S. Typhimurium genome is devoted to the synthesis and uptake of Vitamin B12 and to B12-dependent reactions. In order to understand the requirement from cobalamin synthesis better, we constructed Salmonella serovar Gallinarum, Salmonella serovar Enteritidis and Salmonella serovar Pullorum mutants that are double-defective in cobalamin biosynthesis (ΔcobSΔcbiA). We compared the virulence of these mutants to that of their respective wild type strains and found no impairment in S. Enteritidis ΔcobSΔcbiA and S. Pullorum ΔcobSΔcbiA ability to cause disease in chickens. S. Gallinarum ΔcobSΔcbiA mutant showed attenuated for chickens. We then assessed B12 production by these mutants and their respective wild type strains, as well as S. Typhimurium ΔcobSΔcbiA, and their respective wild type strain. All mutants were unable to produce detectable B12. B12 was detectable in wild type strains, but, S. Gallinarum demonstrated no in vitro cobalamin production. In conclusion, the production of vitamin B12 in vitro differed across the Salmonella serotypes that were tested. Furthermore, the deletion of the cbiA and cobS genes resulted in an alteration in the relationship between the serotype Gallinarum and the birds more stronger than did to the others serotypes. / Mestre

Orientador: Marco Antônio Zanini / Banca: Luiz Antonio de Lima Resende / Banca: Rodrigo Bazan / Banca: Silvana Bommarito Monteiro / Banca: Sthela Zanchetta / Resumo: A deficiência de vitamina B12 no ser humano, frequentemente associada a deficiência de fator intrínseco, produzido no estômago, leva a múltiplas alterações neurológicas e hematológicas. A mais conhecida alteração do sistema nervoso central é a desmielinização e degeneração dos fascículos grácil e cuneiforme da medula cervical. Este trabalho teve por objetivo tentar obter um modelo experimental da afecção, através da gastrectomia total em ratos. Animais foram gastrectomizados, submetidos à suplementação polivitamínica via oral, e 2 a 3 meses depois submetidos a estudos de condução nervosa, com estímulos elétricos na extremidade da cauda, e captação de potenciais elétricos em 3 pontos, com distâncias iguais entre si: próximo ao ânus (potencial caudal), região inferior da coluna (potencial lombar) e região da protuberância cervical (potencial cortical). A velocidade de condução pela região inferior da medula espinhal foi denominada segmento V1, pela região torácica alta e cervical até o córtex, segmento V2. Após obtenções de registros de boa qualidade técnica em 15 gastrectomizados, os segmentos V1 e V2 foram comparados com dados de 20 controles pelo teste "t" de Student para amostras independentes, análise de variância (ANOVA) e teste de comparação múltipla de Tukey. Os animais gastrectomizados apresentaram reduções de velocidades estatisticamente significativas para o segmento V2. Os autores pensam ter conseguido modelo experimental da afecção em ratos, o que abre amplas perspectivas para testes terapêuticos em laboratório, com possibilidades para benefícios futuros ao homem / Abstract: The B12 Vitamin deficiency in man, often associated with deficiency of intrinsic factor produced in the stomach, leading to several neurological and hematological disturbances. The best known alteration of the central nervous system is the demyelination and degeneration of the gracilis and cuneiform fascicles in the cervical spinal cord. This study aimed to try an experimental model of the disease, through total gastrectomy in rats. Animals were gastrectomized, submitted to oral polivitaminic supplementation, and 2-3 months later underwent nerve conduction studies. Electrical stimulation was done on the tail end, and electrical potentials were obtained at 3 points with equal distances from each other: near the anus (tail potential), lower spinal cord (lumbar potential) and the region of cervical spinal cord (cortical potential). The conduction velocity of the lower region of the spinal cord segment was named V1, the high spinal cord to the cortex was named V2 segment. After good quality records from 15 gastrectomized animals, the V1 and V2 segments were compared with data from 20 controls by the "t" Student test for independent samples, analysis of variance (ANOVA) and Tukey's multiple comparison test. Gastrectomized animals presented statistically significant reductions in the velocities for the V2 segment. The authors think they have obtained an experimental model of the disease in rats, which opens up broad prospects for therapeutic tests in the laboratory, with possibilities for future benefits to the man / Mestre