Efficacy of mechanical postconditioning following warm, global ischaemia depends on circulating fatty acid levels in an isolated, working rat heart model

Gebiet: Chirurgie Abstract: OBJECTIVES: – The number of heart transplantations is limited by donor organ availability. Donation after circulatory determination of death (DCDD) could significantly improve graft availability, however, organs undergo warm ischaemia followed by reperfusion, leading to tissue damage. Laboratory studies suggest that mechanical postconditioning [(MPC), brief, intermittent periods of ischaemia at the onset of reperfusion] can limit reperfusion injury, however, clinical translation has been disappointing. We hypothesized that MPC-induced cardioprotection depends on fatty acid levels at reperfusion. – – METHODS: – Experiments were performed with an isolated rat heart model of DCDD. Hearts of male Wistar rats (n = 42) underwent working-mode perfusion for 20 min (baseline), 27 min of global ischaemia and 60 min reperfusion with or without MPC (two cycles of 30 s reperfusion/30 s ischaemia) in the presence or absence of high fat [(HF), 1.2 mM palmitate]. Haemodynamic parameters, necrosis factors and oxygen consumption (O2C) were assessed. Recovery rate was calculated as the value at 60 min reperfusion expressed as a percentage of the mean baseline value. The Kruskal-Wallis test was used to provide an overview of differences between experimental groups, and pairwise comparisons were performed to compare specific time points of interest for parameters with significant overall results. – – RESULTS: – Percent recovery of left ventricular (LV) work [developed pressure (DP)-heart rate product] at 60 min reperfusion was higher in hearts reperfused without fat versus with fat (58 ± 8 vs 23 ± 26%, P < 0.01) in the absence of MPC. In the absence of fat, MPC did not affect post-ischaemic haemodynamic recovery. Among the hearts reperfused with HF, two significantly different subgroups emerged according to recovery of LV work: low recovery (LoR) and high recovery (HiR) subgroups. At 60 min reperfusion, recovery was increased with MPC versus no MPC for LV work (79 ± 6 vs 55 ± 7, respectively, P < 0.05) in HiR subgroups and for DP (40 ± 27 vs 4 ± 2%), dP/dtmax (37 ± 24 vs 5 ± 3%) and dP/dtmin (33 ± 21 vs 5 ± 4%, P < 0.01 for all) in LoR subgroups. – – CONCLUSIONS: – Effects of MPC depend on energy substrate availability, MPC increased recovery of LV work in the presence, but not in the absence, of HF. Controlled reperfusion may be useful for therapeutic strategies aimed at improving post-ischaemic recovery of cardiac DCDD grafts, and ultimately in increasing donor heart availability.

Microbiota depletion promotes browning of white adipose tissue and reduces obesity.

Brown adipose tissue (BAT) promotes a lean and healthy phenotype and improves insulin sensitivity. In response to cold or exercise, brown fat cells also emerge in the white adipose tissue (WAT; also known as beige cells), a process known as browning. Here we show that the development of functional beige fat in the inguinal subcutaneous adipose tissue (ingSAT) and perigonadal visceral adipose tissue (pgVAT) is promoted by the depletion of microbiota either by means of antibiotic treatment or in germ-free mice. This leads to improved glucose tolerance and insulin sensitivity and decreased white fat and adipocyte size in lean mice, obese leptin-deficient (ob/ob) mice and high-fat diet (HFD)-fed mice. Such metabolic improvements are mediated by eosinophil infiltration, enhanced type 2 cytokine signaling and M2 macrophage polarization in the subcutaneous white fat depots of microbiota-depleted animals. The metabolic phenotype and the browning of the subcutaneous fat are impaired by the suppression of type 2 cytokine signaling, and they are reversed by recolonization of the antibiotic-treated or germ-free mice with microbes. These results provide insight into the microbiota-fat signaling axis and beige-fat development in health and metabolic disease.

Psychosocial influences on fruit, vegetable and milk consumption among a cohort of Houston-area 5th graders

Epidemiological evidence suggests that fruit and vegetable intake is negatively associated with the development of several chronic diseases, including heart disease, some cancers and diabetes mellitus. Inadequate consumption of milk during developmental years is associated with osteoporosis. Consumption of fruit, vegetable and milk (FVM) declines from childhood to adolescence. Adolescent eating habits persist into adulthood; thus, understanding psychosocial factors such as self-efficacy, norms and preferences, is important for developing effective interventions. Preferences, one of the most consistent correlates of fruit and vegetable consumption in children and adolescents, may mediate the relationships between self-efficacy and norms and fruit and vegetable consumption. ^ Fifth grade students from one middle school in South Texas were followed for two years. Students completed lunch food records and questionnaires assessing fruit, vegetable and milk self-efficacy and norms and fruit and vegetable preferences. Principal component analyses identified four scales: Fruit Self-Efficacy, Vegetable Self-Efficacy, Fruit and Vegetable Norms, and Milk Influences. Reliability and validity of the four scales and emerging subscales were assessed using Cronbach's alpha and consumption data, respectively. Associations between longitudinal FVM consumption and self-efficacy and norms were tested. Additionally, the influence of preferences on the relationship of self-efficacy, norms and fruit and vegetable consumption was examined. ^ Confirmatory factor analyses confirmed four scales and subscales. Internal consistency and test-retest reliabilities were acceptable. Self-efficacy and norms were related to FVM consumption and changes in fruit and high fat vegetable consumption over the two-year period. While intake over the two-year period differed statistically, eating patterns were stable. Preferences mediated the relation between fruit self-efficacy and FV norms and fruit consumption. ^ In conclusion, self-efficacy and norms about consuming FVM at school appear to influence FVM consumption. Because eating patterns were similar over the two-year period, establishing healthy eating habits in elementary school is important. While FVM self-efficacy and norms influenced FVM consumption in children, only fruit preferences mediated the relationship of fruit consumption, self-efficacy and norms. Since the influences of FVM consumption appear to differ, interventions designed to increase consumption should target these differences and consider the specificity of self-efficacy and norms. ^

Expression study of the atherosclerosis mouse aorta reveals significant disturbance of calcium signaling pathway

Atherosclerosis is a complex disease resulting from interactions of genetic and environmental risk factors leading to heart failure and stroke. Using an atherosclerotic mouse model (ldlr-/-, apobec1-/- designated as LDb), we performed microarray analysis to identify candidate genes and pathways, which are most perturbed in changes in the following risk factors: genetics (control C57BL/6 vs. LDb mice), shearstress (lesion-prone vs. lesion-resistant regions in LDb mice), diet (chow vs. high fat fed LDb mice) and age (2-month-old vs. 8-month old LDb mice). ^ Atherosclerotic lesion quantification and lipid profile studies were performed to assess the disease phenotype. A microarray study was performed on lesion-prone and lesion-resistant regions of each aorta. Briefly, 32 male C57BL/6 and LDb mice (n =16/each) were fed on either chow or high fat diet, sacrificed at 2- and 8-months old, and RNA isolated from the aortic lesion-prone and aortic lesion-resistant segments. Using 64 Affymetrix Murine 430 2.0 chips, we profiled differentially expressed genes with the cut off value of FDR ≤ 0.15 for t-test, and q <0.0001 for the ANOVA. The data were normalized using two normalization methods---invariant probe sets (Loess) and Quantile normalization, the statistical analysis was performed using t-tests and ANOVA, and pathway characterization was done using Pathway Express (Wayne State). The result identified the calcium signaling pathway as the most significant overrepresented pathway, followed by focal adhesion. In the calcium signaling pathway, 56 genes were found to be significantly differentially expressed out of 180 genes listed in the KEGG calcium signaling pathway. Nineteen of these genes were consistently identified by both statistical tests, 11 of which were unique to the test, and 26 were unique to the ANOVA test, using the cutoffs noted above. ^ In conclusion, this finding suggested that hypercholesterolemia drives the disease progression by altering the expression of calcium channels and regulators which subsequently results in cell differentiation, growth, adhesion, cytoskeletal change and death. Clinically, this pathway may serve as an important target for future therapeutic intervention, and thus the calcium signaling pathway may serve as an important target for future diagnostic and therapeutic intervention. ^

Dietary patterns and risk of pancreatic cancer in a hospital-based case-control study

Background. It is important to understand the association between diet and risk of pancreatic cancer in order to better understand the etiology of pancreatic cancer.^ Objectives. Describe the dietary patterns of cases of adenocarcinoma of the pancreas and non-cancer controls and evaluate the odds of having a healthy eating pattern among cases and non-cancer controls.^ Design and Methods. An ongoing hospital-based case-control study was conducted in Houston, Texas from 2000-2008 with 678 pancreatic adenocarcinoma cases and 724 controls. Participants completed a food frequency questionnaire and a risk factor questionnaire. Dietary patterns were derived by principal component analysis and associations between dietary patterns and pancreatic cancer risk were assessed using unconditional logistic regression.^ Results. Two dietary patterns were derived: fruit-vegetable and high fat-meat. There were no statistically significant associations between the fruit-vegetable pattern and pancreatic cancer. An inverse association was seen between the high fat-meat pattern and pancreatic cancer risk when comparing those in the upper intake quintile to those scoring in the lowest quintile after adjusting for demographic and risk factor variables (OR=0.67, p=0.03). In sex-stratified analysis adjusted for demographic and risk factor variables, females scoring in the upper intake quintile of the fruit-vegetable pattern had a 49% lower risk of pancreatic cancer compared to females scoring in the lowest quintile (OR=0.51, p=0.03). An inverse relationship was also seen for the high fat-meat pattern when comparing females in the upper intake quintile to females in the lowest quintile (OR=0.50, p=0.03). In males, neither dietary pattern was significantly associated with pancreatic cancer.^ Conclusions. The current findings for the fruit-vegetable pattern are similar to those of previous studies and support the hypothesis that there is an inverse association between a “healthy” diet (comprised of fruits, vegetables, and whole grains) and risk of having pancreatic cancer (in females only). However, the inverse relationship with the high fat-meat pattern and risk of pancreatic cancer is contrary to other results. Further research on dietary patters and pancreatic cancer risk may lead to better understanding of the etiologic cause of pancreatic cancer.^

DIETARY RELATIONSHIPS TO FATAL BREAST CANCER AMONG SEVENTH-DAY ADVENTISTS

Laboratory experiments in animals, correlational and migrant studies in humans suggest a role for diet in the etiology of breast cancer. Data gathered from individuals via case-control studies are less consistent. Seventh-day Adventist women experience lower mortality from breast cancer than comparable U.S. populations and this decrease is thought to be, at least in part, related to dietary practices (half are vegetarian). In 1960, 25,000 California Seventh-day Adventists completed a questionnaire which included a 21 item food frequency section. Cancer mortality in this population was monitored between 1960 and 1980 and the relationship of high fat food intake and fatal breast cancer was evaluated. Although established risk factors for breast cancer were observed in this population (e.g. age at menarche, age at first pregnancy, age at menopause and obesity) consumption of high fat foods were not observed to exert a strong influence on fatal breast cancer risk. Odds ratios (O.R.) for fatal breast cancer among non-vegetarians was 1.2. Increasing meat consumption bore little relation to risk; O.R. = 1.0, 1.2, 1.1 for consumption categories of none/occasional, 1-3 days/week and 4+ days/week respectively. Nor did the consumption of other high fat foods of animal origin (e.g. butter, cheese, milk, eggs) show any relationship to risk. These results remained unchanged after simultaneously controlling for the effect of other, potentially confounding variables (menstrual characteristics, obesity) via logistic regression analysis. ^

Factores nutricionales que influyen sobre las variables productivas y el desarrollo del tracto gastrointestinal en pollitas comerciales rubias = Nutritional factors affecting productive performance and gastrointestinal tract traits in commercial brown-egg laying pullets

El objetivo general de esta Tesis doctoral fue estudiar la influencia de diversos factores nutricionales sobre los parámetros productivos y el desarrollo del tracto digestivo de pollitas rubias destinadas a la producción de huevos comerciales. Para alcanzar este objetivo se realizaron tres experimentos donde se estudió el cereal principal, el tamaño de partícula del cereal y el nivel de energía y la presentación de los piensos. En el experimento 1 se estudió la influencia del cereal (piensos con enzimas) y la presentación del pienso sobre los parámetros productivos y las características del tracto digestivo en 576 pollitas rubias de 1 a 120 d de edad. De 1 a 45 d de la edad, se utilizaron 4 piensos experimentales organizados de forma factorial con 2 cereales al 50% de inclusión (maíz vs. trigo) y 2 presentaciones del pienso (harina vs. gránulo de 2- mm de diámetro). Cada tratamiento se replicó 6 veces (24 pollitas por réplica). De 46 a 120 d de edad todas las dietas (maíz o trigo) se ofrecieron en harina y por tanto, la única diferencia entre tratamientos fue el cereal utilizado. De 1 a 120 d de edad, las pollitas que recibieron los piensos basados en maíz tuvieron una ganancia de peso vivo (PV) superior (P < 0,05) que las que recibieron los piensos basados en trigo, pero el índice de conversión (IC) fue similar para ambos grupos. De 1 a 45 d de edad, las pollitas alimentadas con gránulo consumieron más pienso (P < 0,001) y tuvieron una ganancia de peso superior (P < 0,001) que las pollitas alimentadas con harina. Gran parte de los efectos beneficiosos de la granulación sobre los parámetros productivos se mantuvieron al final de la prueba (120 d de edad). A los 45 d de edad, el peso relativo de la molleja (PR; g/kg PV) fue superior (P < 0,01) en pollitas alimentadas con maíz que en pollitas alimentadas con trigo. La alimentación en gránulo redujo el PR del tracto gastro intestinal (TGI) y de la molleja (P < 0,001), así como la longitud relativa (LR; cm/kg PV) del intestino delgado (P< 0.01) a ambas edades (45 y 120 d de edad). El tipo de cereal utilizado no afectó al pH del contenido de la molleja a 120 d de edad pero fué inferior (P < 0,01) en las pollitas que recibieron el pienso en harina de 1 a 45 d de la edad que en las que recibieron el pienso en gránulo. Se concluye que el trigo puede substituir al maíz en piensos para pollitas si se acepta una ligera reducción en la ganancia de peso. Asímismo, la alimentación en gránulo de 1 a 45 d de edad aumentó la ganancia de peso a esta edad y al final de la prueba, así como el pH de la molleja a 120 d de edad. La presentación del pienso en gránulo redujo el PR de la molleja y la LR del TGI a 120 d de edad. En el experimento 2 se utilizaron un total de 864 pollitas rubias Hy-Line de 1 d de edad para estudiar la influencia del cereal de la dieta (500 g de maíz o trigo/kg) y el tamaño de partícula del mismo (molienda con molino de martillos con un diámetro de criba de 6, 8, o 10-mm) sobre los parámetros productivos y las características del TGI de 1 a 120 d de edad. Cada uno de los 6 tratamientos se replicó 6 veces (24 pollitas por réplica). De 1 a 45 d de edad, la ganancia de PV aumentó (P< 0,001) y el IC se mejoró (P < 0,05) al reducir el tamaño de partícula del cereal, pero no se observaron diferencias en el periodo crecimiento de 45 a 120 d de edad. A los 45 d de vida, las pollitas alimentadas con maíz tendieron (P < 0,10) a tener un mayor PR del TGI y del proventrículo y una mayor LR del intestino delgado que las pollitas alimentadas con trigo. Asímismo, el PR del TGI a esta edad, aumentó (P < 0,05) a medida que aumentaba el tamaño de partícula del cereal utilizado. A los 120 d de edad, el tratamiento no afectó el PR de ninguno de los órganos del TGI ni al pH de la molleja. Sin embargo, la LR del intestino delgado fue superior (P < 0,05) para las pollitas alimentadas con trigo que para las pollitas alimentadas con maíz. La LR del TGI se redujó (P < 0,05) al aumentar el tamaño de partícula del cereal. Se concluye que el trigo puede incluirse 500 g/kg en piensos de pollitas de 1 a 120 días de edad y que el tamaño de partícula de los cereales afecta el crecimiento de las pollitas durante los primeros 45 d de vida, pero no después. Por lo tanto, se recomienda moler el cereal utilizado al inicio del período de recría (1 a 45 d de edad) con una criba de diámetro igual o inferior a 8 mm. En el experimento 3 se utilizaron un total de 1.152 pollitas rubias Hy-Line de 1 d de edad para estudiar la influencia del nivel de energía de la dieta y la presentación del pienso sobre la productividad y las características del TGI. De 1 a 45 d de edad se utilizaron 6 piensos organizados de forma factorial con 3 concentraciones energéticas (baja: 11,44 MJ; media: 12,05 MJ y alta: 12,66 MJ/kg) y 2 presentaciones del pienso (harina vs. gránulo). De 45 a 120 d todos los piensos experimentales se suministraron en forma de harina y por tanto, la única diferencia entre tratamientos fue el nivel de EMAn utilizado. Cada uno de los 6 tratamientos se replicó 8 veces y la unidad experimental fue la jaula con 24 pollitas. De 1 a 120 d de edad, la ganancia de PV y el IC mejoraron a medida que aumentó la EMAn del pienso (P < 0,001). Las pollitas alimentadas con gránulo de 1 a 45 d de edad comieron mas y tuvieron una ganancia de peso superior (P < 0,001) que las alimentadas con harina. En el global de la prueba, la ganancia de PV fue mayor (P < 0,01) para las pollitas alimentadas con piensos en gránulo. A los 45 d de edad, el PR de todos los segmentos del TGI estudiados fue inferior para las pollitas alimentadas con piensos de alta energía que para las pollitas alimentadas con piensos de media o baja energía. A 120 d de edad, el PR de la molleja fue superior (P < 0,01) para las pollitas alimentadas con piensos de baja energía que con los otros piensos. Sin embargo, la LR del TGI no se vió afectada por el nivel de energía de los piensos. A los 45 d de edad, la alimentación con gránulo redujo el PR del proventrículo (P < 0,05), de la molleja (P < 0,001) y del TGI (P < 0.001), así como la LR del intestino delgado (P < 0,05) y de los ciegos (P < 0,001). A pesar de que las pollitas solo recibieron los piensos en gránulo durante los primeros 45 d de vida, la alimentación con gránulos redujo el PR de la molleja y del proventrículo a 120 d de edad. Se concluye que la alimentación con gránulos durante los primeros 45 d de vida mejora el consumo de pienso y el PV de las pollitas a 120 d de edad. Un aumento del nivel de energía de la dieta de 12,0 a 12,7 MJ/kg mejora los parámetros productivos de 1 a 120 d de edad pero reduce el tamaño del proventrículo y de la molleja. En base de estos resultados concluimos que maíz y trigo con enzimas pueden utilizarse indistintamente en piensos para pollitas de 1 a 120 d de edad con sólo una ligera disminución del PV final con trigo. La granulación y la reducción del tamaño de partícula del cereal del pienso de primera edad (1 a 45 d de vida) y el uso de piensos de alta densidad energética, mejoran los PV a 120 d de edad. Por lo tanto, es recomendable moler los cereales con cribas de no más de 8-mm de diámetro. También, la granulación del pienso y el uso de piensos de alta energía (pobres en fibra bruta) pueden reducir el desarrollo del TGI especialmente de la molleja, lo que puede perjudicar el consumo posterior de pienso durante el inicio del ciclo de puesta. ABSTRACT The general objective of this Thesis was to study the effect of different nutritional factors on productive performance and the development of the gastrointestinal tract (GIT) of commercial brown egg-laying pullets from 1 to 120 d of age. In this respect, the influence of type and particle size of the cereal, and feed form, and energy content of the die,t were studied in 3 experiments. In experiment 1, the influence of the main cereal and feed form of the diet on performance and GIT traits was studied in 576 brown-egg laying pullets from 1 to 120 d of age. From 1 to 45 d of age, 4 diets arranged factorially with 2 cereals (maize vs. wheat) and 2 feed forms (mash vs. pellets) were used. Each treatment was replicated 6 times (24 pullets per replicate). From 46 to 120 d of age, all diets were offered in mash form and therefore, the only difference among diets was the cereal used. Cumulatively, pullets fed the maize diets had higher body weight (BW) gain (P< 0.05) but similar feed conversion ratio (FCR) than pullets fed the wheat diets. From 1 to 45 d of age, pullets fed pellets consumed more feed (P < 0.001) and had higher BW gain (P < 0.001) than pullets fed mash. Most of the beneficial effects of pelleting on productive performance of the birds were still evident at 120 d of age. At 45 d of age, gizzard relative weight (RW; g/kg BW) was higher (P < 0.01) in pullets fed maize than in pullets fed wheat. Feeding pellets reduced the RW of the GIT and the gizzard (P < 0.001) as well as the relative length (RL; cm/kg BW) of the small intestine (SI, P < 0.01) at both ages. The pH of the gizzard contents at 120 d of age was not affected by the main cereal of the diet, but was lower in pullets fed mash from 1 to 45 d of age (P < 0.01) than in pullets fed pellets. We conclude that wheat supplemented with enzymes can be used in substitution of maize in pullet diets with only a slight reduction in BW gain at 120 d of age. Also, feeding pellets from 1 to 45 d of age increased BW gain and pH of the gizzard, and reduced the RW of the gizzard and the RL of the GIT at 120 d of age. In experiment 2, a total of 864 brown-egg laying pullets was used to study the effects of the main cereal of the diet (500 g maize or wheat/kg) and particle size of the cereal (hammer milled to pass through a 6, 8, and 10-mm screen) on growth performance and GIT traits from 1 to 120 d of age. Each of the 6 treatments was replicated 6 times (24 pullets per replicate). Type of cereal did not affect pullet performance at any age. From 1 to 45 d of age, BW gain was increased (P < 0.001) and FCR was improved (P < 0.05) as the particle size of the cereal was reduced, but no effects were observed after this age. At 45 d of age, pullets fed maize tended (P < 0.10) to have a heavier RW of the GIT and proventriculus and a higher relative length (RL, cm/kg BW) of the SI than pullets fed wheat. Also at this age, the RW of the GIT increased (P < 0.05) with increases in particle size of the cereal. At 120 d of age, dietary treatment did not affect the RW of any of the organs studied or gizzard pH, but the RL of the SI was higher (P < 0.05) for pullets fed wheat than for pullets fed maize. Also, the RL of the SI was reduced (P < 0.05) as the particle size of the cereal increased. We conclude that 500 g wheat/kg can be included in pullet feeds from 1 to 120 d of age, and that particle size of the cereal affects pullet performance during the first 45 d of life but not thereafter. Therefore, it is recommended to grind the cereal used in this period with a screen size of no more than 8-mm. In experiment 3, a total of 1,152 one-day-old Hy-Line Brown egg laying pullets were used to study the influence of the energy content of the diet and feed form on productive performance and on several GIT traits. From 1 to 45 d of age, there were 6 diets arranged factorially with 3 concentrations of AMEn (low: 11.66 MJ/kg, medium: 12.05 MJ/kg and high: 12.66 MJ/kg) of the diet and 2 feed forms (mash vs. pellets). From 45 to 120 d all diets were fed in mash form and therefore, the only difference among treatments in this period was the energy content of the diets. Each of the 6 treatments was replicated 8 times and the experimental unit was formed by 24 pullets. Cumulatively, BW gain and FCR improved as the AMEn of the diet increased (P < 0.001). Also, pullets fed pellets from 1 to 45 d of age had higher feed intake and BW gain (P < 0.001) in this period and higher cumulative BW gain (P < 0.01) than pullets fed mash. At 45 d of age, the RWof all the segments of the GIT was lower for pullets fed the high- than for pullets fed the medium- or low- energy diets. At 120 d of age, the RW of the gizzard was higher (P < 0.01) for pullets fed the low energy diets than for pullets fed the other diets. However, the RL of the GIT was not affected by the energy content of the diet. Feeding pellets reduced the RW of the proventriculus (P < 0.05), gizzard (P < 0.001), and GIT (P < 0.001), as well as the RL of the small intestine (P < 0.05) and the ceaca (P < 0.001) at 45 d of age. The effects of feeding pellets on RW of gizzard and proventriculus were still evident at 120 d of age. We concluded that feeding pellets from 1 to 45 d of age improved feed intake and BW of pullets at 120 d of age and that an increase in the energy content of the diet increased pullet performance at all ages but reduced the RW of the proventriculus and gizzard. We conclude that maize and wheat can be used indistinctly in diets for egg laying pullets from 1 to 120 d of age, with only a slight reduction in final BW when wheat is used. Also, particle size of the cereal affects pullet performance during the first 45 d of life but not thereafter. Pelleting of the feeds, and grinding the cereal with a screen size of no more than 8-mm from 1 to 45 d of age, and the use of high density energy diets are recommended in order to achieve adequate target BW at 120 d of age. However, pelleting of the feed, very fine grinding, and the use of high AMEn diets might hinder the development of the GIT, especially that of the gizzard, which might affect feed intake of laying hens especially at the beginning of the production cycle.

Cocaine- and amphetamine-regulated transcript in the rat vagus nerve: A putative mediator of cholecystokinin-induced satiety

Cocaine- and amphetamine-regulated transcript (CART) is widely expressed in the central nervous system. Recent studies have pointed to a role for CART-derived peptides in inhibiting feeding behavior. Although these actions have generally been attributed to hypothalamic CART, it remains to be determined whether additional CART pathways exist that link signals from the gastrointestinal tract to the central control of food intake. In the present study, we have investigated the presence of CART in the rat vagus nerve and nodose ganglion. In the viscerosensory nodose ganglion, half of the neuron profiles expressed CART and its predicted peptide, as determined by in situ hybridization and immunohistochemistry. CART expression was markedly attenuated after vagotomy, but no modulation was observed after food restriction or high-fat regimes. A large proportion of CART-labeled neuron profiles also expressed cholecystokinin A receptor mRNA. CART-peptide-like immunoreactivity was transported in the vagus nerve and found in a dense fiber plexus in the nucleus tractus solitarii. Studies on CART in the spinal somatosensory system revealed strong immunostaining of the dorsal horn but only a small number of stained cell bodies in dorsal root ganglia. The present results suggest that CART-derived peptides are present in vagal afferent neurons sensitive to cholecystokinin, suggesting that the role of these peptides in feeding may be explained partly by mediating postprandial satiety effects of cholecystokinin.

Glucose intolerance caused by a defect in the entero-insular axis: A study in gastric inhibitory polypeptide receptor knockout mice

Mice with a targeted mutation of the gastric inhibitory polypeptide (GIP) receptor gene (GIPR) were generated to determine the role of GIP as a mediator of signals from the gut to pancreatic β cells. GIPR−/− mice have higher blood glucose levels with impaired initial insulin response after oral glucose load. Although blood glucose levels after meal ingestion are not increased by high-fat diet in GIPR+/+ mice because of compensatory higher insulin secretion, they are significantly increased in GIPR−/− mice because of the lack of such enhancement. Accordingly, early insulin secretion mediated by GIP determines glucose tolerance after oral glucose load in vivo, and because GIP plays an important role in the compensatory enhancement of insulin secretion produced by a high insulin demand, a defect in this entero-insular axis may contribute to the pathogenesis of diabetes.

Cholesterol feeding reduces nuclear forms of sterol regulatory element binding proteins in hamster liver

Cholesterol feeding reduces the mRNAs encoding multiple enzymes in the cholesterol biosynthetic pathway and the low density lipoprotein receptor in livers of hamsters. Here we show that cholesterol feeding also reduces the levels of the nuclear NH2-terminal domains of sterol regulatory element binding proteins (SREBPs), which activate transcription of sterol-regulated genes. We show that livers of hamsters, like those of mice and humans, predominantly produce SREBP-2 and the 1c isoform of SREBP-1. Both are produced as membrane-bound precursors that must be proteolyzed to release the transcriptionally active NH2-terminal domains. Diets containing 0.1% to 1.0% cholesterol decreased the amount of nuclear SREBP-1c without affecting the amount of the membrane precursor or its mRNA, suggesting that cholesterol inhibits the proteolytic processing of SREBP-1 in liver as it does in cultured cells. Cholesterol also appeared to reduce the proteolytic processing of SREBP-2. In addition, at high levels of dietary cholesterol the mRNA encoding SREBP-2 declined and the amount of the precursor also fell, suggesting that cholesterol accumulation also may inhibit transcription of the SREBP-2 gene. The high-cholesterol diets reduced the amount of low density lipoprotein receptor mRNA by 30% and produced a more profound 70–90% reduction in mRNAs encoding 3-hydroxy-3-methylglutaryl CoA synthase and reductase. Treatment with lovastatin and Colestipol, which increases hepatic demands for cholesterol, increased the amount of SREBP-2 mRNA as well as the precursor and nuclear forms of the protein. This treatment caused a reciprocal decline in SREBP-1c mRNA and protein. Considered together, these data suggest that SREBPs play important roles in controlling transcription of sterol-regulated genes in liver, as they do in cultured cells.

Proteolytic cleavage product of 30-kDa adipocyte complement-related protein increases fatty acid oxidation in muscle and causes weight loss in mice

Adipocyte complement-related protein (30 kDa) (Acrp30), a secreted protein of unknown function, is exclusively expressed in differentiated adipocytes; its mRNA is decreased in obese humans and mice. Here we describe novel pharmacological properties of the protease-generated globular head domain of Acrp30 (gAcrp30). Acute treatment of mice with gAcrp30 significantly decreased the elevated levels of plasma free fatty acids caused either by administration of a high fat test meal or by i.v. injection of Intralipid. This effect of gAcrp30 was caused, at least in part, by an acute increase in fatty acid oxidation by muscle. As a result, daily administration of a very low dose of gAcrp30 to mice consuming a high-fat/sucrose diet caused profound and sustainable weight reduction without affecting food intake. Thus, gAcrp30 is a novel pharmacological compound that controls energy homeostasis and exerts its effect primarily at the peripheral level.

Perilipin ablation results in a lean mouse with aberrant adipocyte lipolysis, enhanced leptin production, and resistance to diet-induced obesity

Perilipin coats the lipid droplets of adipocytes and is thought to have a role in regulating triacylglycerol hydrolysis. To study the role of perilipin in vivo, we have created a perilipin knockout mouse. Perilipin null (peri−/−) and wild-type (peri+/+) mice consume equal amounts of food, but the adipose tissue mass in the null animals is reduced to ≈30% of that in wild-type animals. Isolated adipocytes of perilipin null mice exhibit elevated basal lipolysis because of the loss of the protective function of perilipin. They also exhibit dramatically attenuated stimulated lipolytic activity, indicating that perilipin is required for maximal lipolytic activity. Plasma leptin concentrations in null animals were greater than expected for the reduced adipose mass. The peri−/− animals have a greater lean body mass and increased metabolic rate but they also show an increased tendency to develop glucose intolerance and peripheral insulin resistance. When fed a high-fat diet, the perilipin null animals are resistant to diet-induced obesity but not to glucose intolerance. The data reveal a major role for perilipin in adipose lipid metabolism and suggest perilipin as a potential target for attacking problems associated with obesity.

Involvement of specific macrophage-lineage cells surrounding arterioles in barrier and scavenger function in brain cortex.

The transport of solutes between blood and brain is regulated by a specific barrier. Capillary endothelial cells of brain are known to mediate barrier function and facilitate transport. Here we report that specific cells surrounding arterioles, known as Mato's fluorescent granular perithelial (FGP) cells or perivascular microglial cells, contribute to the barrier function. Immunohistochemical and in situ hybridization studies indicate that, in normal brain cortex, type I and type II macrophage scavenger receptors are expressed only in FGP/perivascular microglial cells, and surface markers of macrophage lineage are also detected on them. These cells mediate the uptake of macromolecules, including modified low density lipoprotein, horseradish peroxidase, and ferritin injected either into the blood or into the cerebral ventricles. Accumulation of scavenged materials with aging or after the administration of a high-fat diet results in the formation of honeycomb-like foam cells and the narrowing of the lumen of arterioles in the brain cortex. These results indicate involvement of FGP/perivascular microglial cells in the barrier and scavenger functions in the central nervous system.

Participação da conexina 32 na fisiopatogênese da doença hepática gordurosa não alcoólica em camundongos

A doença hepática gordurosa não alcoólica (DHGNA) abrange alterações desde esteatose até esteato-hepatite não alcoólica (EHNA), podendo evoluir para fibrose, cirrose e carcinoma hepatocelular. A DHGNA é considerada a doença hepática mais comum na atualidade e com prevalência mundial alarmante. Esta doença caracteriza-se, basicamente, pela deposição de triglicérides nos hepatócitos, podendo evoluir com inflamação e fibrose, e está intimamente associada com resistência à insulina (RI), diabetes mellitus tipo 2 e obesidade. Os hepatócitos representam as principais células hepáticas e se comunicam através de junções do tipo gap, formadas principalmente por conexina 32 (Cx32). Esta proteína apresenta importante função no controle da homeostase tecidual, regulando processos fisiológicos e tem sido associada como agente protetor na hepatocarcinogênese e outros processos patológicos, porem pouco se sabe sobre sua participação na DHGNA. Sendo assim, o objetivo deste trabalho foi avaliar a participação da Cx32 na fisiopatogênese da DHGNA, utilizando camundongos knockout para Cx32 (Cx32-KO) submetidos a uma dieta hiperlipídica deficiente em colina. Foram analisados dados biométricos, histopatológicos, função hepática, RI, citocinas inflamatórias, adipocinas, estresse oxidativo, peroxidação lipídica e a expressão de genes envolvidos na DHGNA. Os animais Cx32-KO apresentaram maior acumulo de triglicérides hepáticos em relação aos animais selvagens e, consequentemente, maior peso absoluto e relativo do fígado. Adicionalmente, apresentaram maior inflamação hepática demonstrado pela exacerbação da citocina TNF-α e supressão da IL-10, maior dano hepatocelular indicado pelo aumento das enzimas AST e ALT, aumento da peroxidação lipídica e alterações na expressão de genes chaves na fisiopatogênese da DHGNA, como SREBP1c. No entanto, não houve diferença nos marcadores histopatológicos, RI e estresse oxidativo hepático. Por fim, os animais Cx32-KO apresentaram maior produção de leptina e adiponectina no tecido adiposo. Todos esses resultados revelam que a Cx32 pode atuar como um agente protetor ao desenvolvimento da DHGNA, sugerindo seu potencial como novo alvo terapêutico

PGC-1 alfa como regulador inflamatório na esteato-hepatite não-alcoólica

A doença hepática gordurosa não-alcoólica (NAFLD, do inglês) é a manifestação clínica hepática da síndrome metabólica, cuja incidência aumenta consideravelmente em todo o mundo. A NAFLD pode progredir para um estado de esteatohepatite não-alcoólica (NASH, do inglês), caracterizado por inflamação hepatocelular, com ou sem fibrose. Dados na literatura mostram que o coativador-1 alfa do receptor ativado por proliferadores de peroxissoma gama (PGC-1alfa), além de estar envolvido em diversos processos metabólicos, representa uma estratégia terapêutica promissora na modulação da inflamação. Neste projeto investigamos as alterações inflamatórias no fígado induzida por dieta hiperlipídica e o papel do PGC-1alfa nesse processo. Camundongos C57black/6 receberam dieta hiperlipídica contendo 30% de gordura por 10 semanas. O peso dos animais foi avaliado semanalmente. Após a eutanásia, o tecido adiposo intra-abdominal (retroperitoneal e periepididimal) foi coletado e pesado. Analisamos o perfil glicêmico e lipídico sérico e expressão de genes envolvidos no metabolismo glicêmico e lipídico. Avaliou-se também o aspecto histológico e a inflamação do tecido hepático por quantificação das citocinas IL-6, TNF-alfa e IL-1beta. A dieta rica em gordura conduziu a um aumento dos depósitos de gordura intra-abdominal, hiperglicemia e hiperlipidemia. Os animais também apresentavam esteatohepatite, com aumento de citocinas pró-inflamatórias e diminuição na expressão de PGC-1alfa no tecido hepático. O envolvimento do PGC-1alfa na produção de mediadores inflamatórios por hepatócitos foi avaliado em células HepG2 utilizando RNA de interferência (RNAi). O knockdown da expressão de PGC-1alfa causou aumento na expressão e liberação de IL-6 em hepatócitos via aumento na fosforilação de IkBalfa e consequente ativação do NFkB. Portanto, nossos dados mostram que o PGC-1alfa inibe a produção de mediadores inflamatórios (IL-6) em hepatócitos, e fornecem novas evidências das conexões existentes entre as vias metabólicas e imunes