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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
91

Factors regulating urea-nitrogen recycling in ruminants

Doranalli, Kiran 17 January 2011
A series of experiments were conducted to investigate how dietary and ruminal factors regulate urea-N recycling in ruminants. In Experiments 1, 2, and 3, urea-N kinetics were measured using 4-d intra-jugular infusions of [15N15N]-urea. In Experiment 1, the objective was to determine how interactions between dietary ruminally-degradable protein (RDP) level and ruminally-fermentable carbohydrate (RFC) may alter urea-N transfer to the gastrointestinal tract (GIT) and the utilization of this recycled urea-N in rapidly-growing lambs fed high N diets. The dietary factors were: 1) dry-rolled barley (DRB) vs. pelleted barley (PB) as the principal source of RFC; and 2) dietary levels of RDP of 60 vs. 70% (% of CP). Nitrogen intake, fecal and urinary N excretion increased as dietary RDP level increased; however, method of barley processing had no effect on N use. Dietary treatment had no effect on urea-N kinetics; however, endogenous production of urea-N (UER) exceeded N intake. For all diets, 0.669 to 0.742 of UER was recycled to the GIT; however, 0.636 to 0.756 of the GER was returned to the ornithine cycle. In Experiment 2, the objective was to delineate the effects of partial defaunation of the rumen on urea-N kinetics in lambs fed low or high N diets. Treatments were: 1) partial defaunation (PDFAUN) vs. faunation (FAUN); and 2) low (10%, LOW) vs. high (15%, HIGH) dietary CP. Linoleic acid-rich sunflower oil was fed as a partially-defaunating agent. Partial defaunation decreased ruminal NH3-N concentrations. The UER and urinary urea-N excretion (UUE) were lower, and the GER tended to be lower in PDFAUN as compared to FAUN lambs; however, as a proportion of UER, GER was higher and the proportion of recycled urea-N that was utilized for anabolism (i.e., UUA) tended to be higher in PDFAUN lambs. The UER, GER and UUE were higher in lambs fed diet HIGH; however, as a proportion of UER, GER and its anabolic use were higher in lambs fed diet LOW. In Experiment 3, the objective was to delineate how, at similar N intakes, interactions between ruminal partial defaunation and altering dietary RFC may alter urea-N kinetics and N metabolism in lambs. Treatments were: 1) PDFAUN vs. FAUN; and 2) DRB vs. PB. Urinary N excretion was lower and retained N was higher in PDFAUN compared to FAUN lambs. The UER was similar across treatments; however, the GER, expressed as absolute amounts or as a proportion of UER, UUA, and microbial N supply were higher in PDFAUN compared to FAUN lambs. As a proportion of UER, GER was higher, whereas UUE was lower in lambs fed PB compared to those fed DRB. In Experiment 4, the objective was to determine the effects of feeding oscillating dietary CP compared to static dietary CP concentration on N retention and in vitro urea flux across ruminal epithelia. Dietary treatments consisted of a medium CP diet (MEDIUM; 12.8% CP) or diets with oscillating CP content (OSC) fed in two different sequences i.e., 2 d of low CP (9.7% CP) followed by 2 d of high CP (16.1% CP; OSC-HIGH) or vice-versa (OSC-LOW). Ruminal epithelial tissues were collected and mounted in Ussing chambers under short-circuit conditions and the serosal-to-mucosal urea flux (Jsm-urea) was measured using 14C-urea. Although N intake was similar, retained N and microbial N supply were greater in lambs fed the OSC diets compared to those fed the MEDIUM diet. The total Jsm-urea was higher in lambs fed the OSC-LOW compared to those fed the OSC-HIGH diet. Across diets, the addition of phloretin (a known specific inhibitor of facilitative urea transporter-B; UT-B) reduced Jsm-urea; however, phloretin-insensitive Jsm-urea was the predominant route for transepithelial urea transfer. In summary, data presented in this thesis provide new insights that the improved N retention typically observed in defaunated ruminants and in ruminants fed oscillating dietary CP concentrations is partly mediated via increased urea-N recycling to the GIT and utilization of recycled urea-N for anabolic purposes.
92

Factors regulating urea-nitrogen recycling in ruminants

Doranalli, Kiran 17 January 2011 (has links)
A series of experiments were conducted to investigate how dietary and ruminal factors regulate urea-N recycling in ruminants. In Experiments 1, 2, and 3, urea-N kinetics were measured using 4-d intra-jugular infusions of [15N15N]-urea. In Experiment 1, the objective was to determine how interactions between dietary ruminally-degradable protein (RDP) level and ruminally-fermentable carbohydrate (RFC) may alter urea-N transfer to the gastrointestinal tract (GIT) and the utilization of this recycled urea-N in rapidly-growing lambs fed high N diets. The dietary factors were: 1) dry-rolled barley (DRB) vs. pelleted barley (PB) as the principal source of RFC; and 2) dietary levels of RDP of 60 vs. 70% (% of CP). Nitrogen intake, fecal and urinary N excretion increased as dietary RDP level increased; however, method of barley processing had no effect on N use. Dietary treatment had no effect on urea-N kinetics; however, endogenous production of urea-N (UER) exceeded N intake. For all diets, 0.669 to 0.742 of UER was recycled to the GIT; however, 0.636 to 0.756 of the GER was returned to the ornithine cycle. In Experiment 2, the objective was to delineate the effects of partial defaunation of the rumen on urea-N kinetics in lambs fed low or high N diets. Treatments were: 1) partial defaunation (PDFAUN) vs. faunation (FAUN); and 2) low (10%, LOW) vs. high (15%, HIGH) dietary CP. Linoleic acid-rich sunflower oil was fed as a partially-defaunating agent. Partial defaunation decreased ruminal NH3-N concentrations. The UER and urinary urea-N excretion (UUE) were lower, and the GER tended to be lower in PDFAUN as compared to FAUN lambs; however, as a proportion of UER, GER was higher and the proportion of recycled urea-N that was utilized for anabolism (i.e., UUA) tended to be higher in PDFAUN lambs. The UER, GER and UUE were higher in lambs fed diet HIGH; however, as a proportion of UER, GER and its anabolic use were higher in lambs fed diet LOW. In Experiment 3, the objective was to delineate how, at similar N intakes, interactions between ruminal partial defaunation and altering dietary RFC may alter urea-N kinetics and N metabolism in lambs. Treatments were: 1) PDFAUN vs. FAUN; and 2) DRB vs. PB. Urinary N excretion was lower and retained N was higher in PDFAUN compared to FAUN lambs. The UER was similar across treatments; however, the GER, expressed as absolute amounts or as a proportion of UER, UUA, and microbial N supply were higher in PDFAUN compared to FAUN lambs. As a proportion of UER, GER was higher, whereas UUE was lower in lambs fed PB compared to those fed DRB. In Experiment 4, the objective was to determine the effects of feeding oscillating dietary CP compared to static dietary CP concentration on N retention and in vitro urea flux across ruminal epithelia. Dietary treatments consisted of a medium CP diet (MEDIUM; 12.8% CP) or diets with oscillating CP content (OSC) fed in two different sequences i.e., 2 d of low CP (9.7% CP) followed by 2 d of high CP (16.1% CP; OSC-HIGH) or vice-versa (OSC-LOW). Ruminal epithelial tissues were collected and mounted in Ussing chambers under short-circuit conditions and the serosal-to-mucosal urea flux (Jsm-urea) was measured using 14C-urea. Although N intake was similar, retained N and microbial N supply were greater in lambs fed the OSC diets compared to those fed the MEDIUM diet. The total Jsm-urea was higher in lambs fed the OSC-LOW compared to those fed the OSC-HIGH diet. Across diets, the addition of phloretin (a known specific inhibitor of facilitative urea transporter-B; UT-B) reduced Jsm-urea; however, phloretin-insensitive Jsm-urea was the predominant route for transepithelial urea transfer. In summary, data presented in this thesis provide new insights that the improved N retention typically observed in defaunated ruminants and in ruminants fed oscillating dietary CP concentrations is partly mediated via increased urea-N recycling to the GIT and utilization of recycled urea-N for anabolic purposes.
93

Prevalência de coccidiose e correlação com a saúde intestinal de frangos de corte em agroindústrias brasileiras entre os anos de 2012 a 2014 / Coccidiosis prevalence and correlation with intestinal health of broilers in brazilian agricultural industries between the years 2012 and 2014

Gazoni, Fabio Luis 25 September 2015 (has links)
Coccidiosis is a disease caused by protozoa of the genus Eimeria ssp. These protozoa are intracellular parasites of enterocytes that rupture the host cell, causing damage to the intestinal mucosa. The lesions caused by Eimeria reduce nutrient uptake by broilers, affecting their productivity gain, and also represent a portal of entry for other enteropathogens. The aim of this study was to assess the correlation between lesions caused by Eimeria and the prevalence of coccidiosis and other gastrointestinal disorders among broilers reared in Brazil from 2012 to 2014. Intestinal health was evaluated at 82 poultry houses in Brazil, totaling 5,528 birds aged 12 to 40 days. The rearing period was divided into two phases: phase 1 (12 to 21 days) and phase 2 (22 to 40 days). The broilers, at least three per shed, were collected from three different sites. The following gastrointestinal aspects were analyzed in the present study: presence of cell desquamation, excess fluid, excess mucus, ingestion of contaminated litter, thickened intestinal walls, thin intestinal walls, movement of food bolus, abnormal intestinal tonus, Turkish towel appearance, verminosis, and necrotic enteritis. The classification of the scores for gross lesions caused by Eimeria acervulina, Eimeria maxima, and Eimeria tenella followed the method proposed by Johnson & Reid, [8] and the oocyst count of E. maxima (E. maxima micro) in the mucosa was performed under a light microscope at 100X magnification. The statistical analysis of the Pearson correlation coefficient was carried out by the SAS 9.3 software program [16], using a 95% confidence interval. The results of this study revealed that E. acervulina was the most prevalent (mean of 13.5%) species in both rearing stages. Also, there was a positive correlation with thin intestinal walls and abnormal intestinal tonus in phases 1 and 2, as well as a positive correlation with ingestion of contaminated litter in phase 2. The second highest prevalence was that of E. maxima (mean of 6.75%), with a positive correlation with excess mucus, thickened and thin intestinal walls in phase 1, and a positive correlation with cell desquamation, excess fluid, and Turkish towel appearance in phase 2. E. tenella yielded the lowest prevalence rates (mean of 4.35) among the analyzed Eimeria species, showing a positive correlation with excess fluid in phases 1 and 2 and with thickened intestinal walls and lesions caused by E. maxima in phase 2. The microscopic analysis demonstrated that E. maxima was found in 18% of mucosal scrapings in phase 1, which accounts for a subclinical coccidiosis rate of 282.98% compared with clinical coccidiosis. A positive correlation was observed for E. maxima micro between thickened intestinal walls and lesions caused by E. maxima. E. maxima was detected in mucosal scrapings of 29.6% of the broilers in phase 2, accounting for a subclinical coccidiosis rate of 236.37% compared with clinical coccidiosis. E. maxima micro revealed a positive correlation with excess fluid, necrotic enteritis, E. acervulina, E. maxima, and E. tenella in phase 2. The comparison between the rearing periods showed that subclinical coccidiosis affected 64.45% more broilers in phase 2 than in phase 1. In the gross analysis, E. acervulina was the most prevalent species in both rearing periods. A lesion score equal to 1 was the most frequent among all Eimeria species. Subclinical coccidiosis affected a significant number of broilers in the analyzed Brazilian flocks, and was correlated with several factors that reduce intestinal health. It may be concluded that monitoring is of utmost importance to find out the status of intestinal health of poultry. The microscopic detection of E. maxima (mean of 23.8%) is correlated with factors that negatively affect intestinal health. / A coccidiose é uma enfermidade causada por protozoários do gênero Eimeria ssp. Esses são parasitas intracelulares de enterócitos que rompem a célula hospedeira causando lesões na mucosa intestinal. As lesões causadas pelas Eimerias resultam em redução na capacidade de absorção de nutrientes, afetando o ganho produtivo dos frangos de corte, e representam uma porta de entrada para outros enteropatógenos. Sendo assim, o objetivo desse estudo foi analisar a correlação entre as lesões causadas pelas Eimerias, e a prevalência de coccidiose e de demais alterações encontradas no trato gastrointestinal de frangos de corte produzidos, no Brasil, no período de 2012 a 2014. As avaliações da saúde intestinal foram realizadas em 82 integrações de frangos de corte, no Brasil, totalizando 5.528 aves analisadas com idades entre 12 e 40 dias. O período de produção analisado foi dividido em duas fases: 1ª fase (12 aos 21 dias) e 2ª fase (22 aos 40 dias). Os frangos necropsiados foram coletados em três diferentes pontos e no mínimo três aves por galpão. No presente estudo foram analisadas as seguintes alterações do trato gastrintestinal: presença de descamação celular, excesso de fluido, excesso de muco, ingestão de cama, intestino espesso, intestino fino, passagem de alimento, tônus alterado, toalha turca, verminose e enterite necrótica. A definição dos escores macroscópicos de lesão causados pelas Eimeria acervulina, Eimeria maxima, Eimeria tenella seguiram a metodologia de Johnson & Reid [8], e a contagem de oocistos na mucosa para E. maxima (E. maxima micro) foi realizada com auxílio de microscópio óptico com aumento de 100 X. A análise estatística do coeficiente de correlação de Pearson foi feita com o programa SAS 9.3., com intervalo de confiança de 95%. Os resultados desse estudo demonstraram que a espécie E. acervulina foi a que apresentou maior prevalência (média de 13,5%) em ambas as fases de produção avaliadas. Ainda, referente à E. acervulina, observou-se correlação positiva com intestino fino e tônus intestinal alterado na 1ª e 2ª fase, bem como correlação positiva com ingestão de cama apenas na 2ª fase. A segunda maior prevalência foi da espécie E. maxima (média de 6,75%), obteu-se correlação positiva com excesso de muco, intestino espesso e fino na 1a fase e correlação positiva com descamação celular, excesso de fluído e toalha turca na 2a fase avaliada. A E. tenella representou a menor prevalência (média de 4,35) entre as espécies de Eimerias analisadas, apresentando uma correlação positiva na 1ª e 2ª fase com o excesso de fluído e na 2ª fase com o intestino espesso e com lesões de E. maxima. Na avaliação microscópica, a E. maxima esteve presente em 18% dos raspados de mucosa realizados na 1ª fase, o que representa uma coccidiose subclínica de 282,98% com relação a coccidiose clínica. Para a E. maxima micro foi detectada correlação positiva entre os achados com o intestino espesso e com as lesões de E. maxima. Na 2ª fase, E. maxima foi encontrada nos raspados de mucosa de 29,6% das aves, representando uma coccidiose subclínica de 236,37% com relação à coccidiose clínica. A E. maxima micro apresentou na 2ª fase uma correlação positiva com o excesso de fluido, enterite necrótica, E. acervulina, E. maxima e E. tenella. Na análise comparativa entre os períodos, a coccidiose subclínica acometeu 64,45% mais frangos de corte na 2ª fase em relação a 1ª fase. Na avaliação macroscópia de lesões relacionadas à coccidiose, a E. acervulina foi a espécie de maior prevalência em ambas fases de produção. O escore de lesão mais frequente para todas as espécies de Eimerias foi o de grau 1. A coccidiose subclínica acometeu um número expressivo de frangos de corte do plantel brasileiro e foi correlacionada com diversos fatores de diminuição de saúde intestinal. Concluiu-se que o monitoramento é de suma importância para conhecer o status de saúde intestinal dos lotes avícolas. Pois, a E. maxima microscópica está presente (média de 23,8%) com correlação aos fatores que reduzem a saúde intestinal.
94

Cereal Induced Autoimmune Diabetes is Associated with Small Intestinal Inflammation, Downregulated Anti-Inflammatory Innate Immunity and Impaired Pancreatic Homeostasis

Patrick, Christopher January 2014 (has links)
Background: Intestinal inflammation elicited by environmental determinants including dietary proteins and microbes is implicated in type 1 diabetes (T1D) pathogenesis. Also, intrinsic pancreatic abnormalities could precede classic insulitis, contributing to T1D. Materials and Methods: Spontaneous rat T1D models were used for in situ analyses of gut and pancreas to explore novel disease pathways using immunohistochemistry and detailed morphometry, gene expression studies, and molecular screening analyses. Results: In BBdp rats, feeding a cereal diet stimulated T1D under germ-free or specific pathogen-free (SPF) conditions compared with a protective hydrolyzed casein (HC) diet. Cereal-induced T1D was paralleled by increased gut T cell infiltration and TH1-associated pro-inflammatory transcription. HC-fed rats displayed an increased number of anti-inflammatory CD163+ M2 macrophages compared with cereal-fed rats. Cereal-associated promotion of T1D in Lewis diabetes-prone (LEW-DP) rats, a different rat model, similarly featured gut T cell infiltration in conjunction with decreased immunoregulation. The Camp gene was induced in diet-protected HC-fed BBdp rats. Camp encodes the cathelicidin antimicrobial peptide (CAMP), a pleiotropic immunomodulatory host defence factor. Intestinal CAMP was enriched in CD163+ M2 macrophages and could represent a novel marker of these tolerogenic innate immune cells. CAMP expression was also discovered in pancreatic lymph nodes (PLN) and islets, indicating a novel role for this factor in target tissue homeostasis. There was a positive correlation between pancreatic CAMP and total islet number. Also, islet-associated CAMP+ cells were increased in rats with islet inflammation, suggesting upregulation in parallel with insulitis. Exogenous CAMP/LL-37 injections increased the abundance of T1D-protective probiotic bacteria and promoted islet neogenesis in BBdp rats. A prospective partial pancreatectomy (PPx) study was performed to obtain pre-diabetic pancreas biopsies from iii pre-insulitic BBdp rats. The number of endothelium-associated CD68+ macrophages was increased in pre-diabetic pancreata, indicating that perivascular inflammation was an early lesion in the animals. In addition, pre-diabetic pancreata featured enhanced regenerative Reg3a and Reg3b gene expression, indicating abnormal islet expansion preceding insulitis. Conclusions: Small intestinal inflammation paired with deficits in local immunoregulation parallels T1D development. CAMP represents a novel factor in T1D that could have several pleiotropic functions including regulation of commensal microbes, intestinal homeostasis, and pancreatic homeostasis. In addition, target tissue abnormalities precede insulitis and T1D. This research focused on the integrative biology of T1D pathogenesis in spontaneous rat models. This work provides a novel working model that incorporates key roles for gut lumen antigens, intestinal immunity, and the role of islets and altered regenerative capacity in T1D. This research could lead to new therapeutic opportunities for T1D treatment.
95

Molecular Identification and Functional Characteristics of Peptide Transporter 1 (PEPT1) in the Bonnethead Shark (Sphyrna tiburo)

Hart, Hannah 01 January 2015 (has links)
Many elasmobranchs are considered top predators with worldwide distribution, and in general these fish play an important role in the transfer of energy from the lower to the upper trophic levels within the marine ecosystem. Despite this, little research has been done regarding the rates of prey ingestion, digestion, and the processes of energy and nutrient absorption. Specifically understudied is enzymatic digestion within the intestinal brush border, which functions to break down macromolecules into smaller subunits for luminal absorption across the gastrointestinal epithelium. Given their carnivorous diet, the present study sought to expand knowledge on nutrient intake in elasmobranchs by focusing on the uptake of products of protein metabolism. To accomplish this, sequence encoding Peptide Transporter 1 (PepT1), a protein found within the brush border membrane (BBM) of higher vertebrates that is responsible for the translocation and absorption of small peptides released during digestion by luminal and membrane-bound proteases, was molecularly identified in the bonnethead shark (Sphyrna tiburo) using degenerate primers based on conserved portions of known PEPT1 sequences from other vertebrates. Sequence encoding Peptide Transporter 2 (PepT2) was also isolated from the S. tiburo scroll valve intestine using the same methodology. PepT1 was then localized using immunocytochemistry with rabbit polyclonal anti-rat PEPT1 in the esophagus, stomach, duodenum, scroll valve intestine, rectum, and pancreas. Vesicle studies were used to identify the apparent affinity of the transporter, and to quantify the rate of uptake by its H+-dependent cotransporter properties, using 3H-glycylsarcosine as a model dipeptide. The results of this study provide insight into the rate and properties of food passage within S. tiburo, and can lead to future work on topics such as physiological regulation of protein metabolism and absorption and how it may vary in elasmobranchs that exhibit different feeding strategies.
96

Alterações na expressão gênica do tecido gástrico e intestinal de pacientes portadores de diabetes mellitus tipo 2 submetidos à derivação gástrica em Y de Roux / Changes in the gene expression of gastric and intestinal tissues of patients with tpe 2 diabetes mellitus submitted to Roux-en-Y gastric bypass

Priscila Sala Kobal 16 March 2017 (has links)
Em pacientes obesos com diabetes mellitus tipo 2 (DM2), o controle glicêmico pode ser observado em poucos dias após derivação gástrica em Y de Roux (DGYR), antes de ocorrer significativa perda de peso. Dentre os mecanismos propostos para explicar esse benefício metabólico, aqueles que envolvem adaptações gastrintestinais (GI) em resposta às mudanças anatômicas cirúrgicas são os mais consistentes. O presente estudo avaliou a resposta global do transcriptoma gastrintestinal à DGYR em pacientes obesos responsivos à remissão pós-operatória de DM2 e sua correlação com marcadores de homeostase glicêmica. Vinte pacientes adultas obesas com DM2 e candidatas à DGYR foram incluídas, de acordo com critérios específicos. Todas foram submetidas à técnica de DGYR, por laparotomia padronizada e acompanhadas até um ano após DGYR, para classificação dos pacientes responsivos (R) e não responsivos (NR) à remissão de DM2, de acordo com o critério da American Diabetes Association (ADA). Variáveis clínicas e bioquímicas envolvidas na homeostase glicêmica foram avaliadas no pré-operatório e, mensalmente, até 90 dias pós-operatórios. Análises transcriptômicas gastrintestinais foram desenvolvidas por técnica de microarray (Genechip 1.0 ST Array; AffymetrixTM), a partir de RNA obtido de biópsias de estômago corpo alto (ECA), estômago corpo médio (ECM), duodeno, jejuno e íleo, coletadas por enteroscopia de duplo balão (EDB), no período pré-operatório e após 90 dias da DGYR. O método não paramétrico Rank Products (RP) foi utilizado para a seleção dos genes diferencialmente expressos (DEGs). Os DEGs identificados por RP foram submetidos ao programa Ingenuity Pathways Analysis (IPA), para a seleção de genes dentro das seguintes funções relacionadas à fisiopatologia do DM2: 1) Metabolismo dos Carboidratos; 2) Metabolismo Lipídico; 3) Metabolismo dos Aminoácidos, 4) Doenças Metabólicas e 5) Desordem do Sistema Endócrino. DEGs relacionados a essas cinco funções foram submetidos a análises de agrupamento hierárquico de Person, classificação funcional de ontologia gênica (GOslim ontology), análise de enriquecimento funcional (software DAVID, com uso de banco de dados KEGG e Biocarta) e de vias canônicas pelo software IPA. Essas análises foram feitas separadamente em pacientes R e NR. DEGs de interesse foram validados por RT-qPCR usando o sistema de ensaio Taqman® e equipamento 7500 FastTM (Life Technologies). O coeficiente de correlação de Spearman correlacionou variáveis clínicas e bioquímicas com variáveis transcriptômicas. Das 20 pacientes estudadas, 12 foram classificadas como R e 8 como NR. Nas 5 funções de interesse, o número de DEGs encontrado no grupo R foi: 99 (ECA), 26 (ECM), 62 (duodeno), 241 (jejuno) e 63 (íleo). Dentre os DEGs encontrados no duodeno, 8 apresentaram correlação positiva forte a muito forte (coeficiente de Spearman > 0,7) com alterações das concentrações sistêmicas de glicemia ou hemoglobina glicada. Destes, LDLR, MMP1, NPC1L1 e SLC2A5 foram submetidos à análise de RT-qPCR e validados pelo método. De acordo com as análises de enriquecimento funcional e vias canônicas entre grupos (R e NR), apenas a via LXR/RXR, representativa de DEGs encontrados no duodeno de pacientes R, apresentou associação com remissão do DM2. Nos demais segmentos gastrintestinais, essas análises não identificaram vias representativas relacionadas com DM2, embora alguns de seus DEGs estivessem potencialmente envolvidos com marcadores da homeostase glicêmica. Em conclusão, DGYR induziu modificações transcriptômicas gastrintestinais em pacientes obesos com remissão pós-operatória de DM2 e algumas delas foram associadas com marcadores de homeostase glicêmica, principalmente por meio de alterações na via LXR no duodeno / In obese patients with type 2 diabetes mellitus (T2DM), glycemic control is observed within a few days after Roux-en-Y gastric bypass (RYGB), even before significant weight loss. Among the mechanisms proposed to explain this metabolic benefit, those involving gastrointestinal (GI) adaptations in response to surgical anatomical rearreagement are the most consistent. The present study evaluated the global response of the gastrointestinal transcriptome to DGYR in obese patients responsive to postoperative remission of T2DM and its correlation with markers of glycemic homeostasis. Twenty adult obese patients with T2DM diagnosis and candidates to RYGB were included, according to specific criteria. All of them were submitted to standardized laparotomy RYGB surgery and followed until one year after RYGB, for the patients classification as responsive (R) and non-responsive (NR) to T2DM remission according to the American Diabetes Association (ADA) criteria. Clinical and biochemical variables involved in glycemic homeostasis were evaluated at preoperative and monthly up to 90 postoperative days. GI transcriptomic analyzes were performed by microarray technique (Genechip 1.0 ST Array; AffymetrixTM), in RNA samples obtained from biopsies of the stomach cardia (SC), stomach fundus (SF), duodenum, jejunum and ileum, collected by double-balloon enteroscopy (DBE) at pre-postoperative and 90 days after RYGB. The non-parametric Rank Products (RP) method was used for selection of differentially expressed genes (DEGs). DEGs identified by RP were submitted to the Ingenuity Pathways Analysis (IPA) program for its selection within the following functions: 1) Carbohydrate Metabolism; 2) Lipid Metabolism; 3) Amino Acids Metabolism, 4) Metabolic Diseases and 5) Endocrine System Disorder. DEGs related with these five functions were submitted to the analysis of Pearson hierarchical clustering, functional classification of gene ontology (GOslim), functional enrichment (DAVID software, using KEGG and Biocarta database), and canonical pathways (IPA). These analyzes were performed separately in R and NR groups. DEGs of interest were validated by RT-qPCR using Taqman® assays in 7500 FastTM (Life Technologies). The Spearman correlation coefficient correlated clinical and biochemical variables with DEGs. From the 20 patients studied, 12 were classified as R and 8 as NR. In the 5 functions of interest, the number of DEGs found in R group were: 99 (SC), 26 (SF), 62 (duodenum), 241 (jejunum) and 63 (ileum). Among the DEGs found in the duodenum, 8 presented a strong to a very strong positive correlation (Spearman coefficient > 0.7) with systemic concentrations of glycemia or glycated hemoglobin. Of these genes, LDLR, MMP1, NPC1L1 and SLC2A5 were subjected to RT-qPCR analysis and validated by the method. According to the functional enrichment and canonical pathways analyzes between groups (R and NR), only the LXR/RXR pathway, representative of DEGs found in the duodenum of R patients, was associated with T2DM remission. In the other GI segments, these analyzes did not identify representative pathways related to T2DM, although some of their DEGs were potentially involved with markers of glycemic homeostasis. In conclusion, RYGB induced gastrointestinal transcriptomics changes in obese patients with postoperative remission of T2DM and some of these were associated with markers of glycemic homeostasis, mainly by changes in LXR pathway in the duodenum
97

Microencapsulação de Bifidobacterium lactis para aplicação em leites fermentados / Bifidobacterium lactis microencapsulation for fermented milks application

Alcina Maria Liserre 19 August 2005 (has links)
Bifidobacterium spp. são microrganismos probióticos que podem ser incorporados em produtos alimentícios. Entretanto, para que seus efeitos benéficos à saúde humana ocorram, é necessário que o número de células viáveis na hora do consumo seja, no mínimo, 106UFC/g. As bifidobactérias são sensíveis à elevada acidez e, por isso, torna-se necessária a busca por métodos que possam proteger a integridade da célula, sendo um deles a microencapsulação. Em uma primeira etapa do trabalho, Bifidobacterium lactis foi encapsulado em micropartículas de alginato e alginato modificado (alginatoquitosana, alginato-quitosana-sureteric e alginato-quitosana-acryl-eze) e sua sobrevivência e liberação das micropartículas em fluidos simulados do trato gastrintestinal foram mensuradas utilizando-se soluções tampão com pH 1,5, 5,6 e 7,5, na presença e na ausência de pepsina (3g/L), pancreatina (1g/L) e bile (10g/L). A liberação de células das micropartículas teve uma relação direta com o pH do tampão. A microencapsulação aumentou a taxa de sobrevivência de B. lactis, em comparação com células não encapsuladas, em soluções tampão com pH 1,5 sem a presença de enzimas. Em suco gástrico simulado com enzimas digestivas, por outro lado, foi observado que a pepsina proporcionou um efeito protetor sobre as células de B. lactis, e nesse caso, as taxas de sobrevivência do microrganismo estavam diretamente relacionadas com o grau de injúria das células. Em uma segunda etapa do trabalho, leites fermentados com Streptococcus salivarius ssp. thermophilus e Lactobacillus delbrueckii ssp. bulgaricus foram enriquecidos com culturas de Bifidobacterium lactis submetidas a quatro tratamentos diferentes: desidratação em temperatura ambiente, liofilização/congelamento, encapsulação em alginatoquitosana e encapsulação em alginato-quitosana-acryl-eze. A população sobrevivente de B. lactis foi determinada semanalmente no leite fermentado e também após tratamento simulando condições do trato gastrintestinal. Os resultados indicaram que na ausência de pepsina, as populações de B. lactis foram reduzidas drasticamente após o contato com tampão pH 1,5, não sendo possível a detecção de células viáveis livres ou encapsuladas após 120 minutos de teste. A presença de pepsina influenciou positivamente a recuperação de células viáveis de B. lactis em todas as condições testadas, mas as culturas na forma desidratada apresentaram melhores resultados que as culturas microencapsuladas ou liofilizadas. No caso do leite fermentado contendo as células desidratadas, a população de B. lactis, após o tratamento em suco gástrico com enzimas, foi superior à detectada no produto antes desse tratamento. Conclui-se que a microencapsulação não foi eficiente para proteger B. lactis em leite fermentado contra injúrias causadas pelo trato gastrintestinal simulado. / Bifidobacterium spp. are microorganisms that can be added to foods. However, the benefits for the human health occur when the numbers of viable cells in the moment of the consumption is at least 106CFU/g. Bifidobacteria are acid sensitive, and methods to protect cell integrity, such as microencapsulation, are needed. In the first part of the present study, Bifidobacterium lactis was encapsulated in microparticles of alginate and modified alginate (alginate-chitosan, alginate-chitosan-sureteric and alginate-chitosan-acryl-eze) and the survival and release from microparticles in simulated gastrointestinal conditions were measured, using buffers (pH 1.5, 5.6 and 7.5), in the absence and presence of pepsin (3g/L), pancreatin (1g/L) and bile. The release from microparticles presented a direct relationship with pH. When the pH was 1.5 and no enzyme was present, encapsulation improved the survival of B. lactis, when compared to free cells. However, pepsin had a protective effect on B. lactis, and the survival rate was directly related to the cells injury degree. In the second part of the study, fermented milk samples containing Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. Bulgaricus were supplemented with B. lactis submitted to four different treatments: dehydration at room temperature, freeze drying, encapsulation in alginate-chitosan and encapsulation in alginate-chitosaacryl-eze. The number of viable B. lactis cells in the fermented milk was determined weekly and also after treatment with simulated gastrointestinal conditions. Results indicated that in the absence of pepsin, the number of viable cells decreased significantly after contact with buffers (pH 1.5), and no viable cell was detected after 120 minutes. Pepsin improved the recovery of viable cells in the assayed gastric conditions, being the dehydrated cultures more resistant than other cultures. In fermented milk containing the dehydrated cells, the number of viable cells increased after treatment with simulated gastrointestinal fluids. Microencapsulation was not an effective procedure to protect B. lactis in fermented milk against injury caused by the simulated gastrointestinal tract.
98

SMART CAPSULE WITH STIMULI-RESPONSIVE POLYMERS FOR TARGETED SAMPLING FROM THE GASTROINTESTINAL TRACT

Sina Nejati (17029686) 25 September 2023 (has links)
<p dir="ltr">The gastrointestinal (GI) tract and its diverse microbial community play a significant role in overall health, impacting various aspects such as metabolism, physiology, nutrition, and immune function. Disruptions in the gut microbiota have been associated with metabolic diseases, colorectal cancer, diabetes, obesity, inflammatory bowel disease, Alzheimer's disease, and depression. Despite recognizing the importance of the gut microbiota, the interrelationship between microbiota, diet, and disease prevention remains unclear. Current techniques for monitoring the microbiome often rely on fecal samples or invasive endoscopic procedures, limiting the understanding of spatial variations in the gut microbiota and posing invasiveness challenges. To address these limitations, this dissertation focuses on the design and development of an electronic-free smart capsule platform capable of targeted sampling of GI fluid within specific regions of the GI tract. The capsule can be retrieved for subsequent bacterial culture and sequencing analysis. The capsule design is based on stimuli-responsive polymers and superabsorbent hydrogels, chosen for their proven safety, compatibility, and scalability. By leveraging the pH variation across the GI tract, the pH-sensitive polymeric coatings dissolve at the desired region, activating the sampling process. The superabsorbent hydrogel inside the capsule collects the sampled GI fluid and facilitates capsule closure upon completion of sampling. Systematic studies were conducted to identify suitable pH-responsive polymer coatings, superabsorbent hydrogels, and processing conditions that effectively operated within the physiological conditions of the GI tract. The technology's effectiveness and safety were validated through rigorous <i>in vitro</i> and <i>in vivo</i> studies using pig models. These studies demonstrated the potential of the technology for targeted sampling of GI fluid in both small and large intestinal regions, enabling subsequent bacterial culture and gene sequencing analysis. Additionally, the capsule design was enhanced with the integration of a metal tracer, enabling traceability throughout the GI tract using X-ray imaging and portable metal detectors for ambulatory screening. This technology holds promise as a non-invasive tool for studying real-time metabolic and molecular interactions among the host, diet, and microbiota in challenging-to-access GI regions. Its application in clinical studies can provide new insights into diet-host-microbiome interactions and contribute to addressing the burden faced by patients and their families dealing with GI-related diseases.</p>
99

Studies on the Reaction of Dietary Methylglyoxal and Creatine during Simulated Gastrointestinal Digestion and in Human Volunteers

Treibmann, Stephanie, Groß, Julia, Pätzold, Susann, Henle, Thomas 18 April 2024 (has links)
The reactive 1,2-dicarbonyl compound methylglyoxal (MGO) is consumed with food and its concentrations decrease during digestion. In the present paper, the reaction of MGO with creatine, arginine, and lysine during simulated digestion, and its reaction with creatine during the digestion in human volunteers, was studied. Therefore, simulated digestion experiments with a gastric and an intestinal phase were performed. Additionally, an intervention study with 12 subjects consuming MGO-containing Manuka honey and creatine simultaneously or separately was conducted. Derivatization with o-phenylenediamine and HPLC–UV was used to measure MGO, while creatine and glycated amino compounds were analyzed via HPLC–MS/MS. We show that MGO quickly reacts with creatine and arginine, but not lysine, during simulated digestion. Creatine reacts with 56% of MGO to form the hydroimidazolone MG-HCr, and arginine reacted with 4% of MGO to form the hydroimidazolone MG-H1. In the intervention study, urinary MG-HCr excretion is higher in subjects who consumed MGO and creatine simultaneously compared to subjects who ingested the substances separately. This demonstrates that the 1,2-dicarbonyl compound MGO reacts with amino compounds during human digestion, and glycated adducts are formed. These contribute to dietary glycation products consumed, and should be considered in studies investigating their physiological consequences.
100

Intestinal Gene Expression Profiling and Fatty Acid Responses to a High-fat Diet

Cedernaes, Jonathan January 2013 (has links)
The gastrointestinal tract (GIT) regulates nutrient uptake, secretes hormones and has a crucial gut flora and enteric nervous system. Of relevance for these functions are the G protein-coupled receptors (GPCRs) and the solute carriers (SLCs). The Adhesion GPCR subfamily is known to mediate neural development and immune system functioning, whereas SLCs transport e.g. amino acids, fatty acids (FAs) and drugs over membranes. We aimed to comprehensively characterize Adhesion GPCR and SLC gene expression along the rat GIT. Using qPCR we measured expression of 78 SLCs as well as all 30 Adhesion GPCRs in a twelve-segment GIT model. 21 of the Adhesion GPCRs had a widespread (≥5 segments) or ubiquitous (≥11 segments) expression. Restricted expression patterns were characteristic for most group VII members. Of the SLCs, we found the majority (56 %) of these transcripts to be expressed in all GIT segments. SLCs were predominantly found in the absorption-responsible gut regions. Both Adhesion GPCRs and SLCs were widely expressed in the rat GIT, suggesting important roles. The distribution of Adhesion GPCRs defines them as a potential pharmacological target. FAs constitute an important energy source and have been implicated in the worldwide obesity increase. FAs and their ratios – indices for activities of e.g. the desaturase enzymes SCD-1 (SCD-16, 16:1n-7/16:0), D6D (18:3n-6/18:2n-6) and D5D (20:4n-6/20:3n-6) – have been associated with e.g. overall mortality and BMI. We examined whether differences in FAs and their indices in five lipid fractions contributed to obesity susceptibility in rats fed a high fat diet (HFD), and the associations of desaturase indices between lipid fractions in animals on different diets. We found that on a HFD, obesity-prone (OP) rats had a higher SCD-16 index and a lower linoleic acid (LA) proportions in subcutaneous adipose tissue (SAT) than obesity-resistant rats. Desaturase indices were significantly correlated between many of the lipid fractions. The higher SCD-16 may indicate higher SCD-1 activity in SAT in OP rats, and combined with lower LA proportions may provide novel insights into HFD-induced obesity. The associations between desaturase indices show that plasma measurements can serve as proxies for some lipid fractions, but the correlations seem to be affected by diet and weight gain.

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