RESUMO
Environmental enrichment for laboratory animals has come to be viewed as a potential method for improving animal well-being in addition to its original sense as a paradigm for learning how experience molds the brain. It is suggested that the term housing supplementation better describes the wide range of alterations to laboratory animal housing that has been proposed or investigated. Changes in the environments of animals have important effects on brain structure, physiology, and behavior--including recovery from illness and injury--and on which genes are expressed in various organs. Studies are reviewed that show how the brain and other organs respond to environmental change. These data warrant caution that minor cage supplementation intended for improvement of animal well-being may alter important aspects of an animal's physiology and development in a manner not easily predicted from available research. Thus, various forms of housing supplementation, although utilized or even preferred by the animals, may not enhance laboratory animal well-being and may be detrimental to the research for which the laboratory animals are used.
Assuntos
Experimentação Animal , Criação de Animais Domésticos/métodos , Bem-Estar do Animal/legislação & jurisprudência , Animais de Laboratório/fisiologia , Comportamento Animal/fisiologia , Encéfalo/fisiologia , Abrigo para Animais/normas , AnimaisRESUMO
SCOPE: Here we have tested the hypothesis that prebiotic galacto-oligosaccharides (GOS) may enhance mucosal barrier function through direct modulation of goblet cell function. METHODS AND RESULTS: Human adenocarcinoma-derived LS174T cells, which exhibit an intestinal goblet cell-like phenotype, were used to examine the non-prebiotic effects of GOS on goblet cell functions. LS174T cells were treated with GOS, and the expression of goblet cell secretory product genes mucin 2 (MUC2), trefoil factor 3 (TFF3), resistin-like molecule beta (RETNLB) and the Golgi-sulfotransferase genes, carbohydrate (N-acetylglucosamine-6-O) sulfotransferase 5 (CHST5) and galactose-3-O-sulfotransferase 2 (GAL3ST2), was determined by real-time quantitative RT-PCR. In addition, the abundance of CHST5, TFF3 and RETNLB was confirmed by Western blot analysis. Following treatment with GOS for 72 h, the expression of MUC2 was significantly upregulated 2-4-fold, CHST5 and RETNLB, 5-7-fold, and TFF3 2-4-fold. Western blot analysis demonstrated increased abundance of RETNLB, TFF3 and CHST5. Addition of the Th2 cytokine IL-13 along with GOS resulted in synergistic induction of RETNLB and CHST5. IL-8 secretion was not affected by GOS treatment, suggesting that the effects of GOS are not mediated through an inflammatory pathway. CONCLUSION: Collectively, the data indicate that GOS may enhance mucosal barrier function through direct stimulation of intestinal goblet cells.
Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Células Caliciformes/efeitos dos fármacos , Oligossacarídeos/farmacologia , Linhagem Celular/efeitos dos fármacos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Interleucina-13/farmacologia , Interleucina-8/metabolismo , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucina-2/genética , Mucina-2/metabolismo , Oligossacarídeos/química , Peptídeos/genética , Peptídeos/metabolismo , Sulfotransferases/genética , Sulfotransferases/metabolismo , Sulfurtransferases/genética , Sulfurtransferases/metabolismo , Fator Trefoil-3 , Carboidrato SulfotransferasesRESUMO
Rates of colon cancer are much higher in African Americans (65:100,000) than in rural South Africans (<5:100,000). The higher rates are associated with higher animal protein and fat, and lower fibre consumption, higher colonic secondary bile acids, lower colonic short-chain fatty acid quantities and higher mucosal proliferative biomarkers of cancer risk in otherwise healthy middle-aged volunteers. Here we investigate further the role of fat and fibre in this association. We performed 2-week food exchanges in subjects from the same populations, where African Americans were fed a high-fibre, low-fat African-style diet and rural Africans a high-fat, low-fibre western-style diet, under close supervision. In comparison with their usual diets, the food changes resulted in remarkable reciprocal changes in mucosal biomarkers of cancer risk and in aspects of the microbiota and metabolome known to affect cancer risk, best illustrated by increased saccharolytic fermentation and butyrogenesis, and suppressed secondary bile acid synthesis in the African Americans.
Assuntos
Colo/microbiologia , Neoplasias do Colo/etiologia , Dieta Hiperlipídica/efeitos adversos , Fibras na Dieta/estatística & dados numéricos , Mucosa Intestinal , Negro ou Afro-Americano/estatística & dados numéricos , Idoso , Biomarcadores/metabolismo , Colo/metabolismo , Dieta com Restrição de Gorduras , Dieta Hiperlipídica/estatística & dados numéricos , Fezes/química , Voluntários Saudáveis , Humanos , Inflamação/etiologia , Inflamação/metabolismo , Metaboloma , Microbiota , Pessoa de Meia-Idade , População Rural/estatística & dados numéricos , África do Sul , Urina/químicaRESUMO
Colonic gases are among the most tangible features of digestion, yet physicians are typically unable to offer long-term relief from clinical complaints of excessive gas. Studies characterizing colonic gases have linked changes in volume or composition with bowel disorders and shown hydrogen gas (H(2)), methane, hydrogen sulphide, and carbon dioxide to be by-products of the interplay between H(2)-producing fermentative bacteria and H(2) consumers (reductive acetogens, methanogenic archaea and sulphate-reducing bacteria [SRB]). Clinically, H(2) and methane measured in breath can indicate lactose and glucose intolerance, small intestinal bacterial overgrowth and IBS. Methane levels are increased in patients with constipation or IBS. Hydrogen sulphide is a by-product of H(2) metabolism by SRB, which are ubiquitous in the colonic mucosa. Although higher hydrogen sulphide and SRB levels have been detected in patients with IBD, and to a lesser extent in colorectal cancer, this colonic gas might have beneficial effects. Moreover, H(2) has been shown to have antioxidant properties and, in the healthy colon, physiological H(2) concentrations might protect the mucosa from oxidative insults, whereas an impaired H(2) economy might facilitate inflammation or carcinogenesis. Therefore, standardized breath gas measurements combined with ever-improving molecular methodologies could provide novel strategies to prevent, diagnose or manage numerous colonic disorders.
Assuntos
Bactérias/metabolismo , Colo/metabolismo , Colo/microbiologia , Homeostase/fisiologia , Hidrogênio/metabolismo , Bactérias/classificação , Dióxido de Carbono/metabolismo , Doenças do Colo/metabolismo , Doenças do Colo/microbiologia , Neoplasias Colorretais/metabolismo , Humanos , Doenças Inflamatórias Intestinais/metabolismo , Metano/metabolismoRESUMO
Sulfur is both crucial to life and a potential threat to health. While colonic sulfur metabolism mediated by eukaryotic cells is relatively well studied, much less is known about sulfur metabolism within gastrointestinal microbes. Sulfated compounds in the colon are either of inorganic (e.g., sulfates, sulfites) or organic (e.g., dietary amino acids and host mucins) origin. The most extensively studied of the microbes involved in colonic sulfur metabolism are the sulfate-reducing bacteria (SRB), which are common colonic inhabitants. Many other microbial pathways are likely to shape colonic sulfur metabolism as well as the composition and availability of sulfated compounds, and these interactions need to be examined in more detail. Hydrogen sulfide is the sulfur derivative that has attracted the most attention in the context of colonic health, and the extent to which it is detrimental or beneficial remains in debate. Several lines of evidence point to SRB or exogenous hydrogen sulfide as potential players in the etiology of intestinal disorders, inflammatory bowel diseases (IBDs) and colorectal cancer in particular. Generation of hydrogen sulfide via pathways other than dissimilatory sulfate reduction may be as, or more, important than those involving the SRB. We suggest here that a novel axis of research is to assess the effects of hydrogen sulfide in shaping colonic microbiome structure. Clearly, in-depth characterization of the microbial pathways involved in colonic sulfur metabolism is necessary for a better understanding of its contribution to colonic disorders and development of therapeutic strategies.
RESUMO
Reduced susceptibility to sporadic colorectal cancer in native Africans (NA) is correlated with low consumption of animal products and greater microbial production of colonic methane. In this context, two hydrogenotrophic microbial groups are of interest, methanogenic Archaea (MA) utilizing H2 to produce methane and sulfate-reducing bacteria (SRB) generating hydrogen sulfide, which has been linked with chronic inflammatory disorders of the colon. In the present study, stool samples from NA, consuming a diet high in resistant starch and low in animal products, and from African Americans (AA) and European Americans (EA), both consuming a typical Western diet, were examined for genetic diversity and structure of Archaea, MA and SRB communities. In general, a greater proportion of NA than AA and EA harboured the full range of targeted hydrogenotrophic groups. Terminal restriction fragment length polymorphism analysis of 16S rRNA genes and specific functional genes, combined with multivariate statistical analyses, revealed that NA harboured more diverse and different Archaea and MA populations than AA and EA. Also, NA harboured significantly distinct SRB populations compared with AA and EA. Taken together, these data are consistent with diet selecting for distinct hydrogenotrophic microbiota.
RESUMO
Currently, standard protocols for microbial DNA extraction from intestinal tissues do not exist. We assessed the efficiency of a commercial kit with and without mechanical disruption. Better quality DNA was obtained without mechanical disruption. Thus, it appears that bead-beating is not required for efficient microbial DNA extraction from intestinal biopsies.
Assuntos
Fracionamento Celular/métodos , DNA Bacteriano/isolamento & purificação , Análise de Variância , Proteínas de Bactérias/genética , Biópsia , Colo/microbiologia , Contaminação por DNA , Humanos , Íleo/microbiologia , Tipagem Molecular/métodos , Kit de Reagentes para DiagnósticoRESUMO
Decades of research have determined that an animal's brain structure and behavior are molded by experience. Expected experience that plays a critical role in early organization of the brain may be encoded via a process of overproduction of synaptic connections followed by the loss of those that are underutilized during a critical period. However, novel information may be encoded throughout life by the formation of new synapses as the individual animal is exposed to new environmental stimuli. Many laboratory species reared in complex environments or trained to perform complex tasks, regardless of the age when the altered experience is introduced, will exhibit an increase in the number of synapses per neuron as well as other anatomical differences from those reared in standard laboratory housing. Nevertheless, even though increased environmental stimulation may result in more "normal" anatomical and physiological development for that species, there is no conclusive evidence that enriched caging is essential or even that it increases well-being in laboratory rodents.