Effects of early nutrition on the wnt and bone morphogenic protein pathways in the developing intestine

Infant formula is consumed by the majority of infants in the United States for at least part of the first year of life. Infant formula lacks many of the bioactive compounds that are naturally occurring in breast milk. Because of this, there has been an increased interest by the companies that manufacture infant formula to include additives that would potentially allow formula to more closely mimic breast milk activity. One such ingredient currently being added to infant formula is prebiotics. Prebiotics are non-digestible food ingredients that beneficially affect the host by selectively stimulating the growth of specific healthful bacteria in the colon. It is speculated that prebiotics replicate the activity of breast milk oligosaccharides, which through the production of butyrate by intestinal microbiota, may interact with the Wnt/BMP pathways. The Wnt/BMP pathways regulate intestinal stem cells, which determine the growth, development and maintenance of the intestine. Therefore, the objective of this study was to explore the effects that the addition of prebiotics to formula have on the regulation of the Wnt/BMP pathways when fed to neonatal piglets, a model commonly used in the study of infant nutrition. Piglets (n=5) were randomized into sow-reared (SR), fed control formula (F), or fed formula with added prebiotics (F+P). Fructooligosaccharides (FOS) (2 g/L) and polydextrose (PDX) (2 g/L) were chosen as the prebiotics for this study, because this combination had been less studied than other combinations. Ileum and ascending colon were collected at 7 and 14 days-of-age. Dry matter content, pH, and short chain fatty acid (SCFA) content was measured. The mRNA expression of β-catenin, sFRP3, sFRP4, frizzled 6, DKK1 (Wnt pathway), gremlin (BMP pathway), TNF-a, HNF-4α and osteopontin (OPN) was measured by RT-qPCR. Piglets fed the F+P diet had greater acetate concentration and lower pH in the ileum at day 14 and in the colon at day 7 and day 14 than F piglets. Butyrate concentrations were highest in SR with F+P not differing from F in ileum at day 14 and colon at day 7 and day 14. Effects of age were seen in all genes, with the exception of OPN, sFRP-3 and sFRP-4. On day 7, no effect of diet was observed in the ileum, however, mRNA expression of DKK1 and frizzled 6 were greater in F+P than SR (p≤0.05). On day 14, gremlin expression was lower and OPN was greater in the ileum of SR piglets compared to F and F+P. Also on day 14, HNF-4α mRNA expression was greater in both ileum and colon of F+P piglets and sFRP3 mRNA expression was greater in the colon than F or SR . In summary, differences were observed between gene expression of F+P and SR piglet intestines, but the supplementation of 2 g/L scFOS and 2 g/L PDX to formula did not shift expression of genes in the Wnt/BMP pathways to be more similar to SR than F. As the Wnt/BMP pathway is known to exist in a gradient along the crypt-villus axis, with Wnt expression dominating in the crypt region and BMP expression dominating in the villi, it was possible that pooling whole tissue reduced our ability to detect treatment effects that would be concentrated in either region. A method was therefore developed to remove intestinal epithelial cells along the villus-to-crypt axis. Twenty-five-day-old F and SR piglets were euthanized and ileal tissue was collected and placed in a dissociation buffer in a shaking water bath. Exfoliated cells were removed at increasing time points from 5 to 100 minutes in order to remove cells along the villus-to-crypt axis. After the final incubation, remaining mucosal tissue was removed using a sterile glass microscope slide and pooled with the final exfoliated cell isolation. After each cell collection, a section of tissue was fixed in formalin for histomorphological examination. Expression of genes in the Wnt/BMP pathways, along with crypt marker genes (CDK5 and v-myb), were measured in both whole ileal tissue, pooled epithelial cells, and separate epithelial cell isolations from the same piglet. The expression of β-catenin, HNF-4α, TNF-α, TGF-β and the crypt marker v-myb matched the expected villus-to-crypt pattern in cells collected after 10 (incubation 1), 30 (incubation 2) and 60 (incubation 3) minutes. However, expression of expression in cells collected after 100 minutes (incubation 4) was variable, which may be due to the fact that crypt cells were not efficiently removed and the presence of unwanted non-epithelial tissue. Gremlin, OPN, DKK1, sFRP3 and sFRP4 expression was not statistically different along the villus-to-crypt axis. Frizzled 6 and CDK5 did not express as we had predicted, with expression highest towards the villi. In summary, the epithelial cell collection method used was not entirely successful. While much of the gene data suggests that cells were removed along the villus-to-crypt axis through the first three incubations, the last incubation, which involved scraping the tissue, removed non-epithelial components of the mucosa, while leaving the crypts intact. In conclusion, the addition of 2 g/L PDX and 2 g/L scFOS did not cause gene expression of the Wnt/BMP pathways to mirror either F or SR expression. New isolation methods to extract cells along the crypt-villus axis should be considered, including the use of a laser capture microdissection. While this combination of prebiotics did not yield the intended effects, future research should be done on other combinations, such as the inclusion of galactooligosaccharides (GOS), which is commonly added to food products including infant formula.

Produção de enzimas quitosanolíticas utilizando Paenibacillus ehimensis e Paenibacillus chitinolyticus para obtenção de quitoologossacarídeos

The acquisition of oligosaccharides from chitosan has been the subject of several studies in the pharmaceutical, biochemical, food and medical due to functional properties of these compounds. This study aimed to boost its production of chitooligosaccharides (COS) through the optimization of production and characterization of chitosanolytic enzymes secreted by microorganisms Paenibacillus chitinolyticus and Paenibacillus ehimensis, and evaluating the antioxidant potential of the products obtained. In the process of optimizing the production of chitosanase were employed strategies Fractional Factorial Experimental Design and Central Composite Rotatable Design. The results identified the chitosan, peptone and yeast extract as the components that influenced the production of chitosanase by these microorganisms. With the optimization of the culture media was possible to obtain an increase of approximately 8.1 times (from 0.043 to 0.35 U.mL U.mL-1) and 7.6 times (from 0.08 U.mL-1 to 0.61 U.mL-1) in the enzymatic activity of chitosanase produced by P. chitinolyticus and P. ehimensis respectively. Enzyme complexes showed high stability in temperature ranges between 30º and 55º C and pH between 5.0 and 9.0. Has seen the share of organic solvents, divalent ions and other chemical agents on the activity of these enzymes, demonstrating high stability of these crude complexes and dependence of Mn2+. The COS generated showed the ability of DPPH radical scavenging activity, reaching a maximum rate of scavenging of 61% and 39% when they were produced with enzymes of P. ehimensis and P. chitinolyticus respectively. The use of these enzymes in raw form might facilitate its use for industrial applications

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Adubação nitrogenada em cobertura no feijoeiro em sucessão ao milheto solteiro e consorciado com guandu-anão.

Considerando que elevadas quantidades de nitrogênio (N) são requeridas pelo feijoeiro em sucessão a resíduos culturais de gramíneas solteiras e/ou consorciadas com leguminosas, objetivou-se com esta pesquisa avaliar o desempenho agronômico do feijoeiro, cultivar Pérola, sob irrigação, em função de doses de N em cobertura, em sucessão aos cultivos de milheto solteiro e consorciado com guandu-anão. O delineamento experimental foi de blocos casualizados, com quatro repetições, em esquema de parcelas subdivididas. As duas parcelas foram constituídas pelo cultivo do feijoeiro em sucessão aos cultivos de milheto solteiro e consorciado com guandu-anão; e, nas subparcelas, foram aleatorizadas as doses de N (0, 60, 120 e 180 kg ha-1). Avaliou-se a produtividade de fitomassa seca das plantas de cobertura; no feijoeiro avaliou-se: número de vagens por planta, número de grãos por vagem, massa de 100 grãos e produtividade de grãos. O milheto solteiro e milheto consorciado com guandu-anão apresentaram produtividade de fitomassa seca semelhante. Apenas a produtividade de grãos foi afetada pelas culturas antecessoras, sendo superior na sucessão ao consórcio de milheto com guandu-anão. Em relação à adubação nitrogenada, a massa de 100 grãos e a produtividade de grãos aumentaram linearmente com as doses de N em cobertura. A dose de 180 kg ha-1 de N não foi suficiente para promover a máxima expressão do potencial para massa de 100 grãos e produtividade do feijoeiro.

A conduit between epidemiological research and regional health policy

OBJECTIVE: To transform data from a research setting into a format that could be used to support strategies encouraging healthy lifestyle choices and service planning within local government. METHODS: Details of the health status and lifestyle behaviours of the Geelong, Victoria, population were generated independently by the Geelong Osteoporosis Study (GOS), a prospective population-based cohort study. Recent GOS follow-up phases provided evidence about patterns of unhealthy diet, physical inactivity, smoking and harmful alcohol use. These factors are well-recognised modifiable risk factors for chronic disease; the dataset was complemented with prevalence estimates for musculoskeletal disease, obesity, diabetes, cardiovascular disease, asthma and cancer. RESULTS: Data were provided to Healthy Together Geelong in aggregate form according to age, sex and suburb. A population statistics company used the data to project health outcomes by suburb for use by local council. This data exchange served as a conduit between epidemiological research and policy development. CONCLUSION AND IMPLICATIONS: Regional policy makers were informed by local evidence, rather than national or state health survey, thereby optimising potential intervention strategies.

Twelve-month outcomes following surgical repair of the achilles tendon

IntroductionIncidence of Achilles tendon rupture (ATR) has increased over recent years, and debate regarding optimal management has been widely documented. Most papers have focused on surgical success, complications and short term region-specific outcomes. Inconsistent use of standardised outcome measures following surgical ATR repair has made it difficult to evaluate the impact of ATR on a patient’s health status post-surgery, and to compare this to other injury types. This study aimed to report the frequency of surgical repairs of the Achilles tendon over a five-year period within an orthopaedic trauma registry, and to investigate return to work (RTW) status, health status and functional outcomes at 12 months post-surgical repair of the Achilles tendon.MethodsTwo hundred and four adults registered by the Victorian Orthopaedic Trauma Outcomes Registry (VOTOR) who underwent surgical repair of the Achilles tendon between July 2009 and June 2014 were included in this prospective cohort study. The Extended Glasgow Outcome Scale (GOS-E), 3-level European Quality of Life 5 Dimension measure (EQ-5D-3L), and RTW status 12 months following surgical ATR repair were collected through structured telephone interviews conducted by trained interviewers.ResultsAt 12 months, 92% of patients were successfully followed up. Of those working prior to injury, 95% had returned to work. 42% of patients reported a full recovery on the GOS-E scale. The prevalence of problems on the EQ-5D–3 L at 12 months was 0.5% for self-care, 11% for anxiety, 13% for mobility, 16% for activity, and 22% for pain. 16% of patients reported problems with more than one domain. The number of surgical repairs of the Achilles tendon within the VOTOR registry decreased by 68% over the five-year study period.ConclusionsOverall, patients recover well following surgical repair of the Achilles tendon. However, in this study, deficits in function persisted for over half of patients at 12 months post-injury. The decreased incidence of surgical Achilles tendon repair may reflect a change in practice at VOTOR hospitals whereby surgery may be becoming less favoured for initial ATR management.

Low lean tissue mass and physical performance as markers of sarcopenia in older men and women

Background: While declines in muscle mass and function occur in all individuals with advancing age, the extent and rate of decline vary in the general population. We aimed to determine the prevalence of low lean tissue mass combined with poor physical function as an indicator of sarcopenia among older men and women residing in southeastern Australia.

Methods: The study involved men and women aged 60+ years from the Geelong Osteoporosis Study (GOS). Skeletal muscle mass was measured as total lean tissue mass by dual energy x-ray absorptiometry (DXA) and expressed as a percentage of body weight to generate the skeletal mass index (SMI); low lean mass was defined as SMI T-score <-1. Low muscle function was based on performance using “timed up-and-go” scores >10s. Physical activity scores were determined using a validated questionnaire for the elderly and falls were self-reported for the previous year. Associations between sarcopenia, physical activity and falls were determined using multivariable regression techniques.

Results: Among 624 men, 233 had low SMI, 169 had low muscle performance and 81 had both, thus meeting criteria for sarcopenia. Among 436 women, 143 had low SMI, 179 had low muscle performance and 70 had both. A general age-related increase in the observed prevalence of sarcopenia appeared to be driven by an age-related increase in low performance. Sarcopenia was associated with lower physical activity scores. No association was detected between sarcopenia and falls for men but an association was observed for women (age-adjusted OR 1.87, 95% CI 1.11, 3.14).

Conclusion: In our population, the prevalence of sarcopenia was 10.6% (95% CI 7.7, 13.4) for men and 14.5% (95% CI 10.8, 18.3) for women. Men and women with sarcopenia were habitually less active and, for women, sarcopenia was associated with increased likelihood of falls.