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Avalia??o do efeito da megadose de vitamina A no colostro humanoRibeiro, Karla Danielly da Silva 12 June 2007 (has links)
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Previous issue date: 2007-06-12 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The mothers supplementation of vitamin A in the postpartum comes being a measure of intervention sufficiently used in the combat to the vitamin deficiency. The objective of this work was to evaluate the effect of the mother megadose of vitamin A under the levels of retinol in colostrum of postpartum mothers receiving care at the Janu?rio Cicco Maternity School (MEJC), Natal, RN, as well as analyzing the influence of the maternal nutritional status in the reply to this supplementation. The study it was transversal type, with participation of 91 women in labor divided in group had participated of the study have controlled (44 women) and supplemented group (47 women). In the period of the morning blood and milk had been collected (milk 0h). After that a capsule of retinil palmitate of (200 000 UI or 60 mg) was supplied to the supplemented group. Another aliquot of colostro was after gotten 24h of the first collection (milk 24h). Retinol in milk and serum was quantified through the High Pressure Liquid Chromatography. The vitamin ingestion was evaluated by the questionnaire of frequency of alimentary consumption. The levels of serum retinol were 40.6 ? 10.6 and 35.9 ? 10.9 ?g/dL in the groups controlled and supplemented, respectively. The women had presented a satisfactory average ingestion of vitamin (1492,4 ?gRAE/dia), however with high prevalence of inadequate consumption (23%). Average values of retinol in milk 0h had been found and 24h of 93.5 ? 50.3 ?g/dL and 99.1 ? 49.3 ?g/dL has the group controlled group, respectively (p>0.05). After the supplementation had a significant increase in the levels of retinol of the supplemented group, being found values of 102.0 ? 56.0 ?g/dL and 196.1 ? 74.0 ?g/dL for milk 0h and 24h, respectively (p<0.0001). The women in labor presented different answers to the supplementation influenced for the basal levels of retinol in colostrum. It was possible to verify that women with deficient levels of retinol in milk had transferred more retinol to milk 24h than ones with adjusted levels, showing a percentage of reply equivalent to 326.1% and 86.5% of increase, respectively (p< 0.0001). Although the apparent normality found in the serum, the studied women are considered of risk to the development of the vitamin deficiency, and megadose was efficient in first 24h after the supplementation and wakes up with the mechanisms considered for transference of vitamin A to the milk / A suplementa??o materna de vitamina A no p?s-parto ? utilizada como medida de interven??o no combate ? defici?ncia de vitamina A. O objetivo deste trabalho foi avaliar o efeito da megadose de vitamina A sobre os n?veis de retinol no colostro de pu?rperas da Maternidade Escola Janu?rio Cicco (MEJC),Natal-RN, assim como analisar a influ?ncia do estado nutricional materno na resposta a esta suplementa??o. O estudo foi do tipo transversal, com participa??o de 91 parturientes divididas em grupo controle (44 mulheres) e grupo suplementado (47 mulheres). No per?odo da manh? foram coletados sangue e leite (leite 0h). Em seguida uma c?psula de palmitato de retinil (200 000 UI ou 60 mg) foi fornecida ?s parturientes do grupo suplementado. Outra al?quota de colostro foi obtida ap?s 24h da primeira coleta (leite 24h). O retinol no leite e soro foi quantificado utilizando a Cromatografia L?quida de Alta Efici?ncia. A ingest?o de vitamina A foi avaliada pelo question?rio de freq??ncia de consumo alimentar. Os n?veis de retinol s?rico nos grupos controle e suplementado foram 35,9 ? 10,6 e 40,6 ? 10,9 ?g/dL, respectivamente. As mulheres apresentaram uma satisfat?ria ingest?o m?dia de vitamina A (1492,4 ?gRAE/dia), por?m com alta preval?ncia de consumo inadequado (23%). Foram encontrados valores m?dios de retinol no leite 0 e 24h do grupo controle de 99,1 ? 49,3 ?g/dL e 93,5 ? 50,3 ?g/dL (p>0,05) respectivamente. Ap?s a suplementa??o houve um aumento significativo nos n?veis de retinol do grupo suplementado, sendo encontrado valores de 102,0 ? 56,0 ?g/dL e 196,1 ? 74,0 ?g/dL (p<0,0001) para leite de 0 h e 24 h, respectivamente. As parturientes apresentaram diferentes respostas ? megadose. Mulheres com n?veis deficientes de retinol no leite transferiram mais retinol ao leite 24h do que as com n?veis adequados, encontrando um percentual de resposta equivalente a 326,1% e 86,5% de aumento, respectivamente (p< 0,0001). Apesar da aparente normalidade encontrada no soro, as lactantes s?o consideradas de risco ao desenvolvimento da defici?ncia de vitamina A, e a megadose foi eficaz nas primeiras 24h ap?s a suplementa??o e est? de acordo com os mecanismos propostos para transfer?ncia da vitamina A ao leite
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Xilanas de sabugo de milho como agente antioxidante, citot?xico, anticoagulante e imunomoduladorSilveira, Raniere Fagundes de Melo 20 October 2014 (has links)
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Previous issue date: 2014-10-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A prospec??o de polissacar?deos farmacologicamente ativos provenientes de subprodutos agr?colas ainda ? uma pr?tica pouco explorada no meio cient?fico. Dessa forma, este trabalho pretendeu ampliar o conhecimento relacionado ?s atividades farmacol?gicas de polissacar?deos extra?dos do sabugo de milho. A partir da farinha de sabugo de milho, um extrato polissacar?dico foi obtido ao combinar ondas de ultrassom em meio alcalino, e ao final do processo o produto foi denominado de EPSM. Esse extrato foi caracterizado fisico-quimicamente e, atrav?s de ensaios in vitro, foi avaliado como agente antioxidante, citot?xico, anticoagulante e imunomodulator. Os resultados indicaram que o EPSM apresentou significativa a??o quelante de metal, al?m de n?o apresentar efeito t?xico para c?lulas de linhagem normal, mas evidenciar efeito citot?xico contra as c?lulas tumorais HeLa, quando causou a morte celular por apoptose. Em adi??o, outros efeitos farmacol?gicos foram observados, o EPSM diminuiu a produ??o de ?xido n?trico (NO) por macr?fagos ativados, e estendeu o tempo de coagula??o sangu?nea quando avaliado pelo ensaio de APTT. Posteriormente, fra??es metan?licas, etan?licas e cet?nicas foram obtidas a partir do fracionamento dos polissacar?deos presentes no EPSM. Foram obtidas cinco fra??es metan?licas, seis fra??es etan?licas e duas cet?nicas; e todas avaliadas quanto ?s atividades antioxidante, citot?xica, anticoagulante e imunomodulat?ria. Dentre as fra??es, E1.4 exibiu significativo efeito qualante de metal, a??o t?xica em c?lulas HeLa por indu??o da apoptose, reduziu a produ??o de NO por macr?fagos ativados, e ampliou o tempo de coagula??o sangu?nea. Esses resultados sinalizaram que os polissacar?deos farmacologicamente ativos do EPSM estariam presentes na fra??o E1.4. A partir do fracionamento da E1.4 foram obtidas seis subfra??es polissacar?dicas com tamanhos distintos: -3; 3-10; 10-30; 30-50; 50-100 e +100 KDa. Cerca de 80% dos polissacar?deos de E1.4 apresentou tamanho inferior a 10 KDa, e todas as subfra??es apresentaram mais de 61% de a??car em suas composi??es qu?micas. Essas subfra??es exibiram diferentes composi??es monossacar?dicas, mas todas apresentaram xilose. As subfra??es evidenciaram distintos efeitos nos ensaios farmacol?gicos in vitro. As subfra??es de menor tamanho (< 30 KDa) demonstraram maior atividade quelante de metal e maior atividade citot?xica contra c?lulas tumorais. As intermedi?rias (entre 30 e 100 KDa) diminu?ram mais a produ??o de NO por macr?fagos ativados, mas a de maior tamanho (+100 KDa) modulou um maior n?mero de citocinas inflamat?rias, e demonstrou um maior efeito anticoagulante. Portanto, a an?lise conjunta dos resultados indica que os polissacar?deos farmacologicamente ativos do ESPM s?o heteroxilanas e foram concentrados nas subfra??es de E1.4, al?m disso, os efeitos farmacol?gicos dessas heteroxilanas dependem do seu tamanho molecular. / Prospecting pharmacological active polysaccharides from agricultural
byproducts, such as corncobs, is an underexplored practice in the scientific community.
Thus, this work aims to expand knowledge about pharmacological activities of
polysaccharides extracted from corncobs. From corn cob flour a extract was obtained by
ultrasound waves in an alkaline medium, and the end of the process the product was
termed PECC (polysaccharidic extract from corncobs). This extract was physicochemical
characterized and evaluated by in vitro assays as an antioxidant, cytotoxic,
anticoagulant and imunomodulator agent. Results indicated significant activity metal
chelating by PECC, and the use of PECC in cell culture cells showed no toxic effects to
normal cell lines, but toxic action against HeLa tumor cells due promoting cell death by
apoptosis. In addition, other pharmacological effects were observed, the PECC
decreased nitric oxide (NO) production by activated macrophages, and prolonged blood
clotting time through APTT assay. Then methanolic, ethanolic and ketone fractions
were obtained from fractionation of PECC polysaccharides. Five methanolic fractions,
six ethanolic fractions and two ketones were obtained; and all fractions were evaluated
for antioxidant, cytotoxic, anticoagulant, immunomodulatory activities. E1.4 fraction
exhibited significant metal chelating effect, a toxic action to induce apoptosis in HeLa
cells, decreased NO production by activated macrophages, and extended blood clotting
time. These results showed that the PECC pharmacological active polysaccharides
would be present in the fraction E1.4. From fractionation of E1.4 polysaccharide six
subfractions with different sizes were obtained: <3; 3-10; 10-30; 30-50; 50-100 and
>100 KDa. About 80% of E1.4 polysaccharides had lower size to 10 KDa, and all the
subfractions showed over 61% sugar in their chemical compositions. These subfractions
exhibited different monosaccharide compositions, but xylose was presented in all of
them. The subfractions exhibited distinct pharmacological effects in in vitro assays.
Smaller subfractions (<30 KDa) had highest metal chelating activity and greater toxic
action in tumor cells. The intermediate fractions (between 30-100 KDa) decreased more
NO production of activated macrophages, for other side, the larger size (>100 KDa)
modulated a greater number of inflammatory cytokines, and the had greatest
anticoagulant effect. Therefore, when analyzing all the results together it is evident that
the PECC pharmacological polysaccharides are heteroxylans, and were concentrated in
E1.4 fraction, and heteroxilanas pharmacological effects depends on their molecular
size. Thus, corncobs could be used as source from molecules with biotechnology
potential
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Metagenoma de comunidades microbianas expostas ? radia??o natural e a metaisFerreira, Henrique C?sar de Jesus 25 September 2014 (has links)
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Previous issue date: 2014-09-25 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Muitas esp?cies se especializaram em viver nos mais variados ambientes existentes demonstrando a not?vel capacidade de adapta??o do mundo microbiano as mais diversas condi??es f?sico-qu?micas. Ambientes expostos ? radia??o natural e a metais s?o escassos ao redor do mundo, apresentando uma microbiota ainda desconhecida. Com um n?mero total estimado entre 4 e 6 x 1030 microrganismos na terra, estes constituem um enorme pool biol?gico e gen?tico a ser explorado. Abordagens metagen?micas, independentes de cultivo, proporcionam uma nova forma de acesso ao potencial gen?mico de amostras ambientais tornando-se uma importante ferramenta para elucida??o de fun??es ecol?gicas, bem como para identifica??o de novas esp?cies e biomol?culas. Neste trabalho, o material gen?tico ambiental de amostras de solo e ?gua do A?ude Boqueir?o de Parelhas-RN, sob influ?ncia de radia??o natural e da presen?a de metais, foi extra?do, pirosequenciado, e as sequ?ncias geradas foram analisadas atrav?s de programas de bioinform?tica (MG-RAST e STAMP). Perfis taxon?micos comparativos de ambas as amostras mostraram alta abund?ncia do Dom?nio Bacteria, seguida por uma pequena parcela atribu?da aos Dom?nios Eucaryota, Archaea e V?rus. Proteobacteria, Actinobacteria e Bacterioidetes foram os filos que mostraram maior domin?ncia em ambas as amostras. Importantes g?neros e esp?cies associados ? resist?ncia aos agentes estressores encontrados na regi?o foram observados. Sequ?ncias relacionadas ? replica??o e reparo do DNA, ao estresse oxidativo e estresse pelo calor e a resist?ncia a compostos t?xicos foram observadas, mostrando uma importante rela??o entre a microbiota e seu perfil metab?lico, influenciada pelas vari?veis ambientais regionais. Os resultados encontrados neste estudo adicionam valiosos e in?ditos dados sobre a composi??o de comunidades microbianas nestas regi?es. / Many species have specialized to live in the most varied existing
environments showing the remarkable adaptability of the microbial world the
most diverse physicochemical conditions. Environments exposed to natural
radiation and metals are scarce around the world, presenting a microbiota still
unknown. With a total number estimated between 4 and 6 x 1030 microrganisms
on earth, they constitute an enormous biological and genetic pool to be
explored. Metagenomic approach independent of cultivation, provides a new
form to access to the potential genomic environmental samples becoming a
powerful tool for the elucidation of ecological functions, metabolic profiles, as
well as to identify new biomolecules. In this context, the genetic material of
environmental soil and water samples from A?ude Boqueirao Parelhas-RN,
under the influence of natural radiation and the presence of metals, was
extracted, pirosequencing and the generated sequences were analyzed by
bioinformatics programs (MG-RAST and STAMP). Taxonomic comparative
profiles of both samples showed high abundance of Domain Bacteria, followed
by a small portion attributable to Eucaryota Domains, Archaea and Viruses.
Proteobacteria, Actinobacteria and Bacterioidetes phyla showed the greater
dominance in both samples. Important genera and species associated with
resistance to various stressors found in region were observed. Sequences
related to oxidative and heat stress, DNA replication and repair, and resistance
to toxic compounds were observed, suggesting a significant relationship
between the microbiota and their metabolic profile, influenced by regional
environmental variables. The results of this study add valuable and unpublished
data on the composition of microbial communities in these regions
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Express?o heter?loga de biossurfactantes identificados em bibliotecas metagen?micasAra?jo, Sinara Carla da Silva 22 August 2014 (has links)
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Previous issue date: 2014-08-22 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Os microrganismos apresentam uma imensa diversidade gen?tica e est?o presentes em toda biosfera, no entanto cerca de 1% das esp?cies pode ser cultivada por t?cnicas laboratoriais padr?o. A metagen?mica tornou poss?vel o acesso direto ao genoma microbiano derivado de amostras ambientais, utilizando t?cnicas independentes de cultivo. A metodologia permite obter informa??o funcional de prote?nas, assim como a identifica??o de potenciais produtos com interesse biotecnol?gico e de novos recursos biol?gicos industrialmente explor?veis, a exemplo novas solu??es para impactos ambientais. ?reas contaminadas com petr?leo s?o caracterizadas por um grande ac?mulo de hidrocarbonetos e surfactantes s?o utilizados para sua biorremedia??o. Sendo assim, a abordagem metagen?nima foi utilizada para selecionar genes envolvidos no processo de degrada??o e biossurfacta??o de hidrocarbonetos. Em um trabalho anterior, o DNA ambiental (eDNA) foi extra?do de amostras de solo coletadas em duas diferentes ?reas (Caatinga e rio salino) do Rio Grande do Norte (Brasil), as bibliotecas metagen?micas foram constru?das e analisadas funcionalmente. Os clones capazes de degradar o ?leo foram avaliados quanto ? capacidade de sintetizar biossurfactante. O clone foi sequenciado e an?lise da sequencia revelou uma ORF com 897 pb, 298 amino?cidos e prote?na de peso molecular pr?ximo a 34 kDa. A busca por homologia no GenBank revelou similaridade com a sequ?ncia que codifica uma prote?na hipot?tica de representantes da fam?lia Halobacteriaceae, que foram mostradas recentemente como produtoras de biossurfactantes. A presen?a da sequ?ncia codificante inserida e do fen?tipo adquirido foram confirmadas. Primers foram desenhados e suas ORFs amplificadas por PCR. Em seguida, foram subclonadas em vetor de express?o pETDuet-1, contendo uma cauda de histidina, para express?o e posterior purifica??o da prote?na de interesse. Os testes foram realizados para confirma??o da atividade de biossurfactante e degrada??o de hidrocarbonetos e apresentaram resultados positivos. O ensaio de imunodetec??o (western blot) com a utiliza??o do anticorpo monoclonal Anti-His? confirmou a presen?a da prote?na ambiental. Esse estudo foi o primeiro a relatar uma poss?vel prote?na com atividade biossurfactante obtida a partir de uma abordagem metagen?mica. / The microorganisms have a vast genetic diversity and they are present
throughout the biosphere, however, only about 1% of the species can be
cultivated by traditional cultivation techniques. Within this diversity there is a
huge pool genetic and biological being explored. The metagenomics has
enabled direct access to microbial genome derived from environmental samples
using independent methods of cultivation. The methodology enables to obtain
functional information about the proteins, as well as identify potential products
with biotechnological interest and new industrially exploitable biological
resources, such as new solutions to environmental impacts. Oil-contaminated
areas are characterized by a large accumulation of hydrocarbons and
surfactants may be used for bioremediation. Thus, the metagenomic approach
was used in this study in order to select genes involved in the degradation and
hydrocarbon emulsification. In a previous work, the environmental DNA (eDNA)
was extracted from soil samples collected from two different areas (Caatinga
and Saline River) of Rio Grande do Norte (Brazil), the metagenomic libraries
were constructed and functionally analyzed. The clone able to degrade the oil
was evaluated for the ability to synthesize biosurfactants. The sequence
analysis revealed an ORF with 897 bp, 298 amino acids and a protein with
around 34 kDa. The search for homology in GenBank revealed sequence
similarity with a hypothetical protein of representatives Halobacteriaceae family,
who were recently shown as strains producing biosurfactants. The presence of
the inserted coding sequence and the acquired phenotype was confirmed.
Primers were designed and the ORF amplified by PCR. The ORF was
subcloned into pETDuet-1 expression vector for subsequent purification of the
protein of interest containing a histidine tail. The tests performed to confirm the
biosurfactant activity and the ability of hydrocarbon degradation showed positive
results. The immunodetection test (western blot) using the monoclonal AntiHis?
confirmed the presence of the environmental protein. This study was the
first to report a possible protein with biosurfactant activity obtained from a
metagenomic approach
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Determinantes gen?ticos e ambientais das doen?as hipertensivas da gravidezFerreira, Leonardo Capistrano 06 October 2014 (has links)
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Previous issue date: 2014-10-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / O desenvolvimento de doen?as complexas, como a pr?-ecl?mpsia, ?
determinado por fatores ambientais e gen?ticos, al?m da poss?vel intera??o entre
esses fatores. As doen?as hipertensivas da gravidez (DHGs) apresentam um
amplo espectro cl?nico, que pode variar desde pr?-ecl?mpsia leve (hipertens?o e
proteinuria) at? formas mais graves, como a ecl?mpsia (convuls?es) e s?ndrome
HELLP (hem?lise, eleva??o das enzimas hep?ticas e plaquetopenia). O espectro
cl?nico parece estar ligado a diferentes mecanismos patol?gicos. Este trabalho tem
como objetivo identificar fatores (gen?ticos e ambientais) envolvidos no
desenvolvimento das DHGs. Usando uma abordagem caso-controle, selecionamos
um total de 1498 mulheres para os estudos epidemiol?gico e gen?tico, abrangendo
755 gr?vidas normotensas (controle); 518 pr?-ecl?mpsia; 84 eclampsia e 141
HELLP. As mulheres foram genotipados para 18 marcadores distribu?dos em cinco
genes candidatos (FLT1, ACVR2A, ERAP1, ERAP2 e LNPEP). Como resultado
das an?lises dos fatores ambientais, encontramos idade materna, paridade e o
?ndice de massa corporal pr?-gestacional como importantes fatores de risco
associados ?s DHGs. As an?lises gen?ticas mostraram que os genes est?o
associados de maneira fen?tipo-espec?fica: ACVR2A com pr?-ecl?mpsia precoce
(rs1424954, p=0,002); FLT1 com s?ndrome HELLP (rs9513095, p=0,003), e ERAP1
com ecl?mpsia (rs30187, p=0,03). Nossos resultados sugerem que diferentes
mecanismos gen?ticos, juntamente com fatores ambientais espec?ficos, contribuem
na determina??o do espectro cl?nico das DHGs. Al?m disso, o refinamento
fenot?pico parece ser um passo essencial na busca por genes de doen?as
complexas / The development of complex diseases such as preeclampsia are determined by
both environmental and genetic factors, but there is also interaction among these
factors. Preeclampsia is a pregnancy-specific disorder characterized by de-novo
hypertension and proteinuria after 20th week of gestation. There is a broad spectrum
of clinical presentations related to hypertensive disorders of pregnancy (HDP) that
can range from mild preeclampsia to eclampsia (seizures) or HELLP syndrome
(Hemolysis, Elevation of Liver enzymes, Low Platelets). Those clinical outcomes
might be linked to different pathological mechanisms. Our work aims to identify
factors (i.e. genes and environmental) associated with the HDP?s clinical spectrum.
Using a case-control approach, we selected a total of 1498 pregnant women for
epidemiological and genetic studies, encompassing 755 normotensive (control); 518
preeclampsia; 84 eclampsia; and 141 HELLP. Women were genotyped for 18 SNPs
across 5 candidate genes (FLT1, ACVR2A, ERAP1, ERAP2 and LNPEP). For the
environmental factors, we found maternal age, parity status and pre-gestational
body mass index as important risk factors associated with disease. Genes were
associated in a phenotype-specific manner: ACVR2A with early preeclampsia
(rs1424954, p=0.002); FLT1 with HELLP syndrome (rs9513095, p=0.003); and
ERAP1 with eclampsia (rs30187, p=0.03). Our results suggest that different genetic
mechanisms along with specific environmental factors might determine the clinical
spectrum of HDP. In addition, phenotype refinement seems to be an essential step
in the search for complex disease genes
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Perfil imunol?gico associado ? susceptibilidade, resist?ncia e cura na infec??o por Leishmania ( Leishmania ) infantumGalv?o, Joanna Gardel Valverde 10 October 2014 (has links)
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Previous issue date: 2014-10-10 / A leishmaniose visceral (LV) ? end?mica no Brasil, sendo a regi?o nordeste a que apresenta maior incid?ncia, apesar de nos ?ltimos 30 anos ter aumentado o n?mero de casos em outras regi?es do pa?s. A LV na Am?rica Latina ? resultante da infec??o por Leishmania infantum. No entanto, nem todas as pessoas infectadas desenvolvem doen?a; na verdade, a maioria apresenta resolu??o espont?nea da infec??o sem sintomas caracter?sticos de LV. Tradicionalmente a avalia??o do perfil imunol?gico tem sido realizada utilizando a reestimula??o de c?lulas mononucleares de sangue perif?rico em cultura. Esses estudos revelaram que pacientes com LV apresentavam inibi??o tanto da prolifera??o linfocit?ria quanto da resposta pr?-inflamat?ria anti-Leishmania. O presente trabalho teve por objetivo avaliar a resposta imune na LV sintom?tica, na cura p?s-tratamento e assintom?tica. Para isso, analisamos caracter?sticas imunofenot?picas relacionadas ? ativa??o, Treg e mem?ria de linf?citos, por citometria de fluxo, assim como a produ??o de citocinas ex vivo ou em cultura de sangue total. Para os volunt?rios com LV, foi realizado um estudo longitudinal com reavalia??o imunol?gicas aos 4 e 14 meses ap?s a cura cl?nica. O grupo controle incluiu pessoas provenientes de regi?o end?mica, sendo dividido em 2 grupos: Controle Positivo, formado por pessoas que apresentaram sorologia e?ou PCR anti-Leishmania positivos e Controle Negativo formado por pessoas com sorologia e PCR anti-Leishmania negativos. Durante a LV os linf?citos CD4 apresentam um maior perfil de ativa??o e mem?ria, al?m de serem maiores produtores de citocinas em cultura, quando comparado aos linf?citos CD8, contudo essa ativa??o n?o ? Leishmania espec?fica, visto que ocorreu tanto em aus?ncia (CD4+CD25+:10,60%, CD8+CD25+:5,36%; p<0,0001) quanto em presen?a de ant?genos sol?veis de Leishmania (SLA), (CD4+CD25+: 10,20%, CD8+CD25+:3,28%; p=0,0003). H? ativa??o de linf?citos durante a LV (CD4+CD69+: 4,9%), quando comparado aos grupos Controle Positivo (CD4+CD69+: 1,96% p=0,0045) e Negativo (CD4+CD69+: 1,35% p=0,0062), mas esta ativa??o tamb?m n?o ? Leishmania espec?fica. O perfil de ativa??o linfocit?ria permanece elevado mesmo 14 meses ap?s o fim do tratamento, por?m ap?s a cura a ativa??o ? Leishmania espec?fica (CD4+CD25+ em aus?ncia de SLA: 8,44%, presen?a de SLA: 10,70% p=0,0279). Linf?citos CD8+CD25+ foram capazes de produzir IFN-? em presen?a de ant?geno tanto em Controles Positivos (aus?ncia de SLA: 5,17%, presen?a de SLA: 9,52% p=0,0391) como em LV curados (Curado 4 meses: aus?ncia de SLA: 3,90%, presen?a SLA: 10,70% p=0,0098). C?lulas presentes no sangue total de pessoas com LV ativa s?o capazes de produzir IFN-? em resposta ao SLA (IFN-? em aus?ncia de SLA: 3,61 pg?mL e em presen?a de SLA: 44,26 pg?mL; p=0,0020), assim como LV recuperado (IFN-? em aus?ncia de SLA: 2,29 pg?mL e presen?a de SLA: 139,80 pg?mL; p=0,0005). Contudo o elevado n?vel de IL-10 parece estar inibindo a atividade pro-inflamat?ria de IFN-? e TNF-? em pacientes na fase sintom?tica. Contrariamente as demais citocinas pro-inflamat?rias, a cultura de sangue total do grupo LV ativa n?o apresentou produ??o de IL-2 Leishmania espec?fica (em aus?ncia de SLA: 2,42 pg?mL e em presen?a de SLA: 2,56 pg?mL). Com base nesses dados n?s conclu?mos que a restaura??o da ativa??o de linf?citos e a diminui??o da produ??o de IL-10, Leishmania espec?fica, est?o relacionados ? um perfil imunol?gico protetor. / Visceral Leishmaniasis (VL) is endemic in Brazil and the northeast region had the
highest incidence
of the disease
, despite, in the last 30 years,
it
has spread
to all
geographic regions of the
country.
Leishmania infantum
is the
m
ain
etiological agent
of VL in Latin America, Europe and North Africa.
However,
not all infected individuals
develop the disease; in fact, the majority present spontaneous re
solution of infection
without
symptoms. The evaluation of the immunological profil
e has been mostly
conducted
stimulating, with
Leishmania
spp. antigen, peripheral blood mononuclear
cells
isolated from subjects with VL. These studies showed that VL patients had an
inhibition of both, lymphocyte proliferation and proinflammatory response
to
Leishmania
spp. antigen. Our study aimed to evaluate the immune response in active
LV, cured post treatment and asymptomatic infection. To reach this aim, we analyzed
immunophenotypic features related to activation, Treg and memory lymphocytes, by
flow
cytometry, as well as, evaluation of cytokine production, in
ex vivo
or in whole
blood culture. In
active
VL volunteers, a longitu
dinal study was
conducted with
reassessment at 4 and 14 months after clinical cure.
The control group included
individuals th
at live
d
in endemic region and were
either
Positive Control, consisting
of individuals with
positive anti
-
L
eishmania
spp.
serology and/or
positive
PCR
for
Leishmania
?
spp.
and Negative Control composed by individuals with
negative anti
-
Leishmania antibodie
s
serology and
negative
PCR
for Leishmania
. During VL, CD4
lymphocytes showed greater activation and memory profile
s
and
were
the
major
source of cytokines in culture when compared to CD8 lymphocytes
, and these were
not
Leishmania
specific. There
were
act
ivated lymphocytes during VL (CD4
+
CD69
+
:4.9%) when compared to control groups, Positive (CD4
+
CD69
+
:1.96%,
p=0.0045) and Negative (CD4
+
CD69
+
:1.35%, p=0.006), on the other hand, this was
non
-
specific activation. The lymphocyte activation profile remain
ed
el
evated even 14
months post treatmen
t. A
fter
clinical
cure
,
the activation
was
Leishmania
specific
(CD4
+
CD25
+
absence of SLA:
8.4%, and presence of SLA:
10.7% p=0.0279).
CD8
+
CD25
+
lymphocytes
were
able to produce
Leishmania
specific IFN
-
? in both,
Positive
Controls (absence of SLA 5.2% and presence of SLA: 9.5%, p=0.0391) and
Cured 4 month (absence of SLA: 3.9%; presence of SLA: 10.7% p=0.0098). Whole
blood culture cells, of VL patients,
were
able to produce IFN
-
?, by SLA stimulation
(absence of SLA: 28.0 pg
?mL, and presence: 44.3 pg?mL p=0.0020) as well as
recovered groups (absence of SLA 2.3 pg?mL and presence of SLA 139.8 pg?mL,
p=0.0005). However, the high level of IL
-
10 seem
ed
to inhibit pro
-
inflammatory
activity of IFN
-
? and TNF
-
?
during symptomatic dis
ease
.
Unlike other pro
-
inflammatory cytokines, active VL group d
id
not produce
Leishmania
specific IL
-
2
(absence of SLA 2.4 pg?mL and presence of SLA: 2.6 pg?mL). Based on these data
we conclude that the restoration of lymphocyte activation and decreased i
n IL
-
10
Leishmania
specific production
were
related to a protective immune profile.
|
207 |
Otimiza??o da extra??o de polissacar?deos sulfatados da alga vermelha Gracilaria birdiae e an?lise da atividade anticoagulante e antioxidanteFidelis, Gabriel Pereira 12 December 2014 (has links)
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Previous issue date: 2014-12-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Polissacar?deos sulfatados (PS) da alga vermelha comest?vel Gracilaria birdiae
foram obtidos atrav?s de cinco diferentes condi??es de extra??o (GB1: ?gua;
GB1p: ?gua/prote?lise; GB1s: ?gua/sonica??o; GB1sp:
?gua/sonica??o/prote?lise; GB2s: NaOH/sonica??o; GB2sp:
NaOH/sonica??o/prote?lise). O rendimento em massa seca (g) das extra??es
aumentou na seguinte ordem: GB2sp>GB1sp>GB1p>GB2s>GB1s>GB1. A
quantidade de PS extra?dos por cada condi??o, em contrapartida, aumentou
em sequencia diferente: GB2sp>GB1p>GB1>GB1sp>GB1s>GB2s.
Espectroscopia de infravermelho e eletroforese em gel de poliacrilamida em
agarose demonstram que todas as condi??es extra?ram o mesmo
polissacar?deo sulfatado. Al?m disso, com base nos dados referentes ?
composi??o monossacar?dica de cada condi??o de extra??o pode-se pressupor
que o uso de sonica??o fez com que se obtivesse outros polissacar?deos al?m
dos PS. No teste de tempo de protrombina (PT), o qual avalia a via extr?nseca
da cascata de coagula??o, nenhuma das amostras apresentou atividade
anticoagulante at? uma concentra??o m?xima de 0,1 mg/mL. Por outro lado, no
teste de tempo de tromboplastina parcial ativada (aPTT), cuja via da cascata de
coagula??o avaliada ? a intr?nseca, todas os extratos de PS tiveram atividade
positiva, com exce??o da condi??o GB2sp. A atividade anticoagulante no teste
de aPTT diminuiu na seguinte sequencia:
GB1sp>GB2sp>GB1p>GB1>GB1s>GB2s. O teste de capacidade antioxidante
total (CAT) dos polissacar?deos sulfatados tamb?m sofreu influ?ncia das
varia??es de acordo com condi??o de extra??o, visto que com Gb2s e Gb1
foram obtidos os menores valores de atividade quando comparado ?s demais
condi??es de extra??o. No presente trabalho, conclui-se que as condi??es de
extra??o de PS influenciam as atividades biol?gicas e na composi??o qu?mica
dos PS obtidos e que portanto, o uso de uma condi??o em especial pode
favorecer a obten??o de PS com uma atividade que se deseja em detrimento
de outras. De acordo com os dados obtidos pode-se afirmar que
NaOH/sonica??o/prote?lise foi a melhor condi??o para obter PS com atividade
anticoagulante e antioxidante em Gracilaria birdiae / The sulfated polysaccharides (SP) from the edible red seaweed Gracilaria
birdiae were obtained using five different condition extraction (GB1: Water;
GB1p: Water/proteolysis; GB1s: Water/sonication; GB1sp:
Water/sonication/proteolysis; GB2s: NaOH/sonication; GB2sp:
NaOH/sonication/proteolysis. The yield (g) increased in the following order
GB2sp>GB1sp>GB1p>GB2s>GB1s>GB1. However, the amount of SP
extracted increased in different way GB2sp>GB1p>GB1>GB1sp>GB1s>GB2s.
Infrared and electrophoresis analysis showed that all conditions extracted the
same SP. In addition, monosaccharide composition showed that ultrasound
promotes the extraction of other polysaccharides than SP. In the prothrombin
time (PT) test, which evaluates the extrinsic coagulation pathway, none of the
samples showed anticoagulant activity. While in the activated partial
thromboplastin time (aPTT) test, which evaluates the intrinsic coagulation
pathway, all samples showed anticoagulant activity, except GB2s. The aPTT
activity decreased in the order of GB1sp>GB2sp>GB1p>GB1>GB1s>GB2s.
Total capacity antioxidant (TCA) of the SP was also affected by condition
extraction, since GB2s and GB1 showed lower activity in comparison to the
other conditions. In conclusion, the conditions of SP extraction influence their
biological activities and chemical composition. The data showed
NaOH/sonication/proteolysis was the best condition to extract anticoagulant and
antioxidant SPs from Gracilaria birdiae.
|
208 |
Caracteriza??o de um hom?logo de HINT1 em cana-de-a??car encontrado em bibliotecas subtrativas de CDNA para flora??oSousa, Isabel Andrade Lopes de 27 February 2015 (has links)
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Previous issue date: 2015-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A cana-de-a??car ? uma planta monocotiled?nea cultivada em regi?es tropicais e
subtropicais, sendo o Brasil o maior produtor mundial. Apesar de sua import?ncia
econ?mica, muito pouco ? conhecido molecularmente sobre o processo de flora??o
em cana-de-a??car. Este processo fisiol?gico pode promover uma perda de at? 60%
na produtividade de a??car ou bioetanol. Desta forma, este trabalho teve como
objetivo caracterizar um gene hom?logo ao gene que codifica a prote?na HINT1,
identificado anteriormente em bibliotecas subtrativas de flora??o. An?lises
gen?micas da estrutura g?nica e regi?o promotora permitiram observar que existem
pelo menos dois genes distintos hom?logos ? HINT em cana-de-a??car. An?lises de
bioinform?tica mostraram a conserva??o do dom?nio proteico caracter?stico da
superfam?lia HIT e indicam uma rela??o filogen?tica associada ? localiza??o celular.
Al?m disso, foi observada uma poss?vel rela??o com as prote?nas da fam?lia
SBTILISIN-like por meio das informa??es dispon?veis em interatomas. Isto sugere
que o gene HINT de cana-de-a??car pode estar relacionado ao desenvolvimento
vegetal, havendo v?rias possibilidades de intera??es para a regula??o do processo
de indu??o floral, pois as sequ?ncias presentes nas regi?es regulat?rias indicam que
a express?o diferencial de HINT seria relacionada tanto com fatores clim?ticos
presentes na regi?o Nordeste do Brasil quanto ? estresse bi?tico e fito-horm?nios.
Al?m disso, os fen?tipos relacionados ao florescimento tardio indicam que a
influ?ncia de HINT pode ocorrer devido ao ac?mulo de produtos na sua atua??o
enzim?tica. Por estas caracter?sticas este gene pode ser utilizado como um
marcador na sele??o de novos cultivares. / The sugarcane is a monocot plant grown in tropical and subtropical regions, with
Brazil being the largest producer. Despite its economic importance, little is known
about the molecular flowering process in sugarcane. This physiological process can
promote a loss up to 60% in sugar or bioethanol. Thus, this work had as objective
characterize a HINT1 homologous gene previously identified in subtractive libraries
of flowering. Genomic analysis of gene and promoter region structure allowed the
observation that there are at least two distinct genes homologous to HINT on
sugarcane. Bioinformatics analyses showed the conservation of the characteristic
protein domain of HIT superfamily and indicate a phylogenetic relationship
associated to cell location. Moreover, a possible relation with the SBTILISIN-like
protein family through the information available in interatomas was observed. This
suggests that the HINT gene of sugarcane can be related to plant development,
there are several possibilities of interactions in the regulation of floral induction
process, because the sequences present in regulatory regions indicate that
differential expression of HINT was related to with climatic factors in the Northeast
region of Brazil as well as to biotic stress and phytohormones. Furthermore, the
sugarcane phenotypes indicate that the influence of HINT may happen due to
product accumulation of its enzymatic activity. For these characteristics this gene can
be used as a marker in the selection of new varieties.
|
209 |
Mol?culas coestimulat?rias na leishmaniose visceralRodrigues Neto, Jo?o Firmino 27 April 2015 (has links)
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Previous issue date: 2015-04-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / A leishmaniose visceral (LV) ? uma doen?a end?mica em muitos pa?ses,
incluindo o Brasil. O protozo?rio Leishmania infantum ? o agente etiol?gico da
LV, sendo transmitido pela picada das f?meas dos flebotom?neos, durante o
repasto sangu?neo. A maioria dos indiv?duos quando expostos ao parasita n?o
desenvolvem a doen?a, pois apresentam um predom?nio da resposta celular
Th1. Aqueles que desenvolvem doen?a, apresentam sinais como febre, perda de
peso, hepatoesplenomegalia e um comprometimento da resposta imune celular,
espec?fica a ant?genos de Leishmania. N?s avaliamos se essa anergia,
observada durante a doen?a ativa, poderia estar associada com altera??es nas
mol?culas coestimulat?rias de linf?citos T ou em seus ligantes em mon?citos
CD14+. H? aumento na porcentagem de CTLA-4 em linf?citos T CD4+ (p=0,001)
e ICOS em linf?citos T CD4+ e CD8+ (p=0,002 para CD4+ e p=0,003 para
CD8+) ap?s est?mulo por ant?geno sol?vel de Leishmania (SLA) na LV
sintom?tica, e que h? maior porcentagem dessas mol?culas ex vivo, quando
comparados indiv?duos sintom?ticos aos recuperados (p=0,04 para CTLA-4 em
CD4+, e p=0,001 para ICOS em CD4+ e p=0,026 para CD8+). Al?m disso,
encontramos uma maior express?o dos genes CTLA-4, OX-40 e ICOS, durante
a LV ativa. As mol?culas CD40, CD80, CD86, HLA-DR e ICOSL, n?o sofrem
altera??o durante a doen?a. H? produ??o de IFN-? por c?lulas de sangue
perif?rico, ap?s est?mulo por SLA, em indiv?duos sintom?ticos; no entanto, h?
diminui??o na raz?o entre IFN-?/IL-10, com aumento desta ap?s a cura. A
observa??o do comprometimento de algumas vias de mol?culas coestimulat?rias
poderia diminuir a capacidade microbicida dos fag?citos, durante a leishmaniose
visceral sintom?tica, podendo facilitar a sobreviv?ncia e a prolifera??o do
parasita. / Visceral leishmaniasis (VL) is endemic in many countries, including Brazil. The
protozoan Leishmania infantum, is the etiological agent of VL, and is transmitted
by the bite of female sandflies during the blood meal. The majority of subjects
when exposed to the parasite do not develop the disease, because of
development of Th1 cellular responses. Those who have develop signs of VL
such as fever, weight loss, hepatosplenomegaly, have impairment of the cellular
immune response, specific to the Leishmania antigens. We evaluated whether
the specififc anergy during symptomatic VL, may be associated with changes in T
cells costimulatory molecules or their ligands in CD14+ monocytes. There is an
increase in CTLA-4 porcentage on CD4+ T lymphocytes (p=0.001) and ICOS on
CD4+ and CD8+ T cells (p=0.002 to CD4+ and p=0.003 to CD8+), after
stimulation by soluble Leishmania antigen (SLA) during active visceral
leishmaniasis, and that there is a higher percentage of these molecules ex vivo,
when comparing symptomatic to recovered individuals (p=0.04 to CTLA-4 in
CD4+, and p=0.001 to ICOS in CD4+ and p=0.026 to CD8+). Moreover, we
found a high gene expression of CTLA-4, OX-40 and ICOS during active VL.
CD40, CD80, CD86, HLA-DR and ICOSL molecules do not suffer changes during
disease. There is IFN-? production by the peripheral blood cells, after SLA
stimulation, by peripheral blood cells in symptomatic subjects; however, there is a
decrease of the ratio IFN-?/IL-10, which is reversed after clinical recovery. The
impairment of some costimulatory molecules pathways during symptomatic VL
could inhibit the ability of phagocytes to kill Leishmania and could facilitate their
survival and the proliferation inside macrophages.
|
210 |
Avalia??o da suplementa??o de vitamina E sobre a concentra??o de alfa-tocoferol no leite materno em mulheres com partos prematurosPires, Jeane Franco 05 May 2015 (has links)
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Previous issue date: 2015-05-05 / O termo vitamina E refere-se a um grupo de oito compostos moleculares que
diferem em estrutura e biodisponibilidade, sendo o RRR-alfa-tocoferol a forma mais
ativa biologicamente. A composi??o de vitamina E no leite materno sofre varia??es
ao longo da lacta??o, sendo o leite colostro mais rico neste micronutriente quando
comparado ao leite de transi??o e maduro. Os rec?m-nascidos, especialmente os
prematuros s?o mais suscept?veis a defici?ncia de vitamina E e para prevenir os
danos causados por esta defici?ncia tem sido proposta a suplementa??o do neonato
com este micronutriente, por?m, n?o existe consenso para realiza??o desta
interven??o. Assim, a suplementa??o materna com RRR-alfa-tocoferol no p?s-parto
pode ser uma boa alternativa para tentar elevar os n?veis de alfa-tocoferol no leite
materno e, consequentemente, fornecer ao rec?m-nascido prematuro quantidades
adequadas de vitamina E. Este estudo objetivou avaliar o efeito da suplementa??o
com 400 UI de acetato de RRR-alfa-tocoferol em mulheres com partos prematuros,
sobre a concentra??o de alfa-tocoferol no leite materno colostro, transi??o e maduro.
Participaram do estudo 89 pu?rperas adultas saud?veis, que foram distribu?das no
grupo controle (n = 51) e grupo suplementado (n = 38). Foram coletadas amostras
de sangue e leite colostro logo ap?s o parto (leite 0h), vinte e quatro horas ap?s,
nova al?quota de leite colostro foi coletado (leite 24h). O leite de transi??o e maduro
foram coletados em sete dias (leite 7d) e trinta dias (leite 30d) ap?s o parto,
respectivamente. A suplementa??o no grupo suplementado foi realizada ap?s a
coleta de sangue e leite 0h. As an?lises de alfa-tocoferol foram realizadas por
cromatografia l?quida de alta efici?ncia. Valores s?ricos de alfa-tocoferol menores
que 516 ?g/dL foram considerados indicativos de defici?ncia nutricional. A
concentra??o m?dia de alfa-tocoferol no soro das parturientes do grupo controle foi
1159,8 ? 292,4 ?g/dL e do grupo suplementado foi 1128,3 ? 407,2 ?g/dL (p = 0,281).
Todas as pu?rperas apresentaram estado nutricional em vitamina E adequado. Em
ambos os grupos, foi poss?vel observar que a concentra??o de vitamina E no leite
colostro foi maior em rela??o ao leite de transi??o e maduro. No grupo
suplementado, a concentra??o de alfa-tocoferol no leite 24h aumentou em 60% ap?s
a suplementa??o, passando de 1339,3 ? 414,2 ?g/dL (leite 0h) para 2234,7 ? 997,3
?g/dL (leite 24h). Enquanto que o grupo controle os valores no colostro 0h e colostro
24h foram semelhantes (p = 0,681). No leite de transi??o do grupo controle o valor
de alfa-tocoferol foi 875,3 ? 292,4 ?g/dL e no grupo suplementado 1352,8 ? 542,3
?g/dL, com aumento de 35% no grupo suplementado em rela??o ao controle (p <
0,001). No leite maduro as concentra??es de alfa-tocoferol entre o grupo controle
(426,6 ? 187,5 ?g/dL) e suplementado (416,4 ? 214,2 ?g/dL) foram semelhantes (p =
0,853). Apenas o leite 24h do grupo suplementado atendeu o requerimento
nutricional de alfa-tocoferol (4 mg/dia) do rec?m-nascido. Tais resultados evidenciam
que o transporte deste micronutriente para o leite ocorre de maneira controlada e
limitada. Dessa forma, a suplementa??o materna com vitamina E eleva a
concentra??o de alfa-tocoferol no leite colostro e de transi??o e n?o influencia a
concentra??o no leite maduro. Apenas o aumento no leite colostro foi suficiente para
atingir o requerimento nutricional do rec?m-nascido prematuro. / The term vitamin E refers to a group of eight molecular compounds which
differ in structure and bioavailability, and the RRR-alpha-tocopherol more biologically
active form. The composition of vitamin E in breast milk undergoes variations during
lactation, colostrum and milk richer in this micronutrient compared to transitional and
mature milk. Newborns, especially premature infants are more susceptible to vitamin
E deficiency and to prevent the damage caused by this deficiency has been
proposed supplementation of neonates with this micronutrient, however, there is no
consensus to carry out this intervention. Thus, maternal supplementation with RRRalpha-tocopherol
in the postpartum period can be a good alternative to try to raise the
alpha-tocopherol levels in breast milk and therefore provide the premature newborn
adequate amounts of vitamin E. This study to evaluate the effect of supplementation
with 400 UI acetate RRR-alpha-tocopherol in women with premature births, on the
concentration of alpha-tocopherol in breast milk colostrum, transitional and mature.
The study included 89 healthy adult women were enrolled in the control group (n =
51) and supplemented group (n = 38). Blood samples were collected and milk
colostrum soon after birth (0h milk) twenty-four hours, new rate of colostrum milk was
collected (24h milk). The transitional and mature milk were collected in seven days
(7d milk) and thirty days (30d milk) after delivery, respectively. Supplementation in
the supplemented group was held after the collection of blood and 0h milk. The
alpha-tocopherol analyzes were performed by high-performance liquid
chromatography. Serum levels of alpha-tocopherol less than 516 ?g/dL were
considered indicative of nutritional deficiency. The average concentration of alphatocopherol
in the serum of the control group mothers was 1159.8 ? 292.4 ?g/dL and
the supplemented group was 1128.3 ? 407.2 ?g/dL (p = 0.281). All women had
nutritional status in vitamin E suitable. In both groups, it was observed that the
concentration of vitamin E in colostrum milk was higher compared to transitional and
mature milk. In the supplemented group, the concentration of alpha-tocopherol in the
milk increased 60 % after supplementation, from 1339.3 ? 414.2 ?g/dL (0h milk) to
2234.7 ? 997.3 ?g/dL (24h milk). While the control group values in colostrum 0h and
colostrum 24h were similar (p = 0.681). In the control group the follow-on milk alphatocopherol
value was 875.3 ? 292.4 ?g/dL and in the group supplemented 1352.8 ?
542.3 ?g/dL, an increase of 35% in the supplemented group compared to control (p
<0.001). In mature milk alpha-tocopherol concentrations between the control group
(426.6 ? 187.5 ?g/dL) and supplemented (416.4 ? 214.2 ?g/dL) were similar (p =
0.853). Only 24h milk supplemented group answered the nutritional requirement of
alpha-tocopherol (4 mg/day) of the newborn. These results show that the transport of
this micronutrient for milk occurs in a controlled and limited way. Thus, the native
vitamin E supplementation increases the concentration of alpha-tocopherol in
colostrum and milk and transition does not influence the concentration in mature milk.
Only the increase in colostrum milk was sufficient to meet the nutritional requirement
of premature newborns.
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