Adherence of Uropathogenic Escherichia Coli in Dog Urine After Consumption of Food Supplemented with Cranberry (Vaccinium Macrocarpon).

Introduction: Escherichia coli is the most common pathogen isolated from the urine of dogs with urinary tract infections (UTIs). While there are many studies in humans investigating the potential for the prevention of UTIs by dietary consumption of cranberry, few analogous studies have been carried out in dogs. Material and Methods: Eight dogs, four male and four female, were successively fed two diets, first a control without cranberry, and then the second diet containing cranberry extracts. Naturally excreted urine was collected on the tenth day after the start of each diet for 24 h and used for bacterial growth. Madin-Darby canine kidney cell adherence by the uropathogenic E. coli G1473 strain expressing type 1 pili and positive for P pili and haemolysin gene markers was quantified after growth in urine samples. Results: Significant reductions in bacterial adherence to MDCK cells (from -16.5 to -73.4%, P < 0.05) were observed in the four females but not in the males after consumption of the cranberry extracts compared to the same animals consuming the control diet. Conclusion: Dietary supplementation with cranberry may provide some degree of protection to female dogs against adhesion of uropathogenic E. coli to urinary epithelial cells.

n-3 PUFA prevent metabolic disturbances associated with obesity and improve endothelial function in golden Syrian hamsters fed with a high-fat diet.

Glucose intolerance and dyslipidaemia are independent risk factors for endothelium dysfunction and CVD. The aim of the present study was to analyse the preventive effect of n-3 PUFA (EPA and DHA) on lipid and carbohydrate disturbances and endothelial dysfunction. Three groups of adult hamsters were studied for 20 weeks: (1) control diet (Control); (2) high-fat diet (HF); (3) high-fat diet enriched with n-3 PUFA (HFn-3) groups. The increase in body weight and fat mass in the HF compared to the Control group (P < 0.05) was not found in the HFn-3 group. Muscle TAG content was similar in the Control and HF groups, but significantly lower in the HFn-3 group (P = 0.008). Glucose tolerance was impaired in the HF compared to the Control group, but this impairment was prevented by n-3 PUFA in the HFn-3 group (P < 0.001). Plasma TAG and cholesterol were higher in the HF group compared to the Control group (P < 0.001), but lower in the HFn-3 group compared to the HF group (P < 0.001). HDL-cholesterol was lower in the HFn-3 group compared to the Control and HF groups (P < 0.0005). Hepatic secretion of TAG was lower in the HFn-3 group compared to the HF group (P < 0.005), but did not differ from the Control group. Hepatic gene expression of sterol regulatory element-binding protein-1c, diacylglycerol O-acyltransferase 2 and stearyl CoA desaturase 1 was lower in the HFn-3 group, whereas carnitine palmitoyl transferase 1 and scavenger receptor class B type 1 expression was higher (P < 0.05). In adipocytes and adipose macrophages, PPARγ and TNFα expression was higher in the HF and HFn-3 groups compared to the Control group. Endothelium relaxation was higher in the HFn-3 (P < 0.001) than in the HF and Control groups, and was correlated with glucose intolerance (P = 0.03) and cholesterol (P = 0.0003). In conclusion, n-3 PUFA prevent some metabolic disturbances induced by high-fat diet and improve endothelial function in hamsters.

Beneficial effects of a prescription home-prepared diet and of zucchini on urine calcium oxalate supersaturation and urinary parameters in adult cats.

OBJECTIVES: From the authors' experience, the consumption of a balanced prescription home-prepared diet that includes zucchini (courgette) benefits cats with recurrent urolithiasis, but there is no published evidence to support this. The aim was to study the effects on urinary parameters of (1) a balanced prescription home-prepared diet containing zucchini, and (2) the addition of zucchini to a dry food, compared with two commercial therapeutic diets. METHODS: Eight healthy cats were included in a Latin-square designed protocol. Five diets were evaluated: two commercial diets, designed for cats with urinary disorders, one high-moisture (U-WET) and one high-sodium dry (U-DRY); one home-prepared diet (HOME); one commercial dry food for adult maintenance (DRY); and DRY given together with 10 g of zucchini per kg body weight (DRY-Zuc). After a 7-day adaptation period, urine was collected and daily food and water intakes were assessed for 12 days. Urinary parameters, and relative supersaturation (RSS) for calcium oxalate (CaOx) and struvite, were determined. Data underwent repeated measures ANOVA analysis. RESULTS: The digestibility of energy, dry matter, protein and fat was highest with the HOME diet. CaOx RSS was lowest in cats eating the HOME diet, but not significantly different from the U-WET or U-DRY diets. CaOx RSS was lower in cats eating the DRY-Zuc diet than in cats eating the DRY diet. Struvite RSS did not differ significantly among groups. CONCLUSIONS AND RELEVANCE: This study shows that a balanced prescription home-prepared diet was safe and allowed a very low urinary CaOx RSS. It also showed that adding zucchini to dry food lowered the urine CaOx RSS.

Diet-induced dyslipidemia impairs reverse cholesterol transport in hamsters.

BACKGROUND: Reverse cholesterol transport (RCT) is an anti-atherogenic process by which cholesterol is effluxed from peripheral tissues by high-density lipoprotein (HDL) and returned to the liver for excretion into the bile and faeces. Dyslipidemia is thought to impair RCT through higher triglyceride-rich lipoprotein (TRL), low HDL-cholesterol and higher activity of cholesteryl ester transfer protein (CETP), which transfers cholesteryl esters from HDL to TRL for further hepatic uptake. As CETP pathway would represent a major route in human RCT, we therefore investigated whether diet-induced dyslipidemia impairs RCT in hamster, a CETP-expressing species. MATERIALS AND METHODS: Golden Syrian hamsters were fed a chow or chow+0·3% cholesterol diet over 4 weeks. Biochemical parameters and in vivo VLDL-triglycerides secretion (Triton WR-1339 injection) were then measured. In vitro macrophage cholesterol efflux was measured, and in vivo macrophage-to-faeces RCT was also assessed after an intraperitoneal injection of (3) H-cholesterol-labelled hamster primary macrophages. RESULTS: Cholesterol-enriched diet increased plasma total cholesterol (144%), triglycerides (101%), VLDL-triglycerides secretion (175%), CETP activity (44%) and reduced HDL-cholesterol/total cholesterol ratio by 20% (P < 0·01 vs. chow). Cholesterol-enriched diet significantly increased hepatic total cholesterol and triglycerides by 459 and 118% and increased aortic total cholesterol content by 304%. In vitro cholesterol efflux from macrophages to plasma was significantly reduced by 25% with plasma from cholesterol-fed hamsters. In vivo RCT experiments showed a significant 75% reduction of macrophage-derived cholesterol faecal excretion in cholesterol-fed hamsters. CONCLUSIONS: Overall, these data demonstrate that diet-induced dyslipidemia severely impairs in vivo RCT in hamsters.

Liver X receptor activation promotes macrophage-to-feces reverse cholesterol transport in a dyslipidemic hamster model.

Liver X receptor (LXR) activation promotes reverse cholesterol transport (RCT) in rodents but has major side effects (increased triglycerides and LDL-cholesterol levels) in species expressing cholesteryl ester transfer protein (CETP). In the face of dyslipidemia, it remains unclear whether LXR activation stimulates RCT in CETP species. We therefore used a hamster model made dyslipidemic with a 0.3% cholesterol diet and treated with vehicle or LXR agonist GW3965 (30 mg/kg bid) over 10 days. To investigate RCT, radiolabeled (3)H-cholesterol macrophages or (3)H-cholesteryl oleate-HDL were then injected to measure plasma and feces radioactivity over 72 or 48 h, respectively. The cholesterol-enriched diet increased VLDL-triglycerides and total cholesterol levels in all lipoprotein fractions and strongly increased liver lipids. Overall, GW3965 failed to improve both dyslipidemia and liver steatosis. However, after (3)H-cholesterol labeled macrophage injection, GW3965 treatment significantly increased the (3)H-tracer appearance by 30% in plasma over 72 h, while fecal (3)H-cholesterol excretion increased by 156% (P < 0.001). After (3)H-cholesteryl oleate-HDL injection, GW3965 increased HDL-derived cholesterol fecal excretion by 64% (P < 0.01 vs. vehicle), while plasma fractional catabolic rate remained unchanged. Despite no beneficial effect on dyslipidemia, LXR activation promotes macrophage-to-feces RCT in dyslipidemic hamsters. These results emphasize the use of species with a more human-like lipoprotein metabolism for drug profiling.

Effects of High-Fat Diet at Two Energetic Levels on Fecal Microbiota, Colonic Barrier, and Metabolic Parameters in Dogs.

Increased consumption of energy-rich foods is a key factor in overweight, obesity, and associated metabolic disorders. This would be, at least in part, related to microbiota disturbance. In rodent models of obesity, microbiota disruption has been associated with alteration of the intestinal barrier, endotoxemia, inflammation grade, and insulin sensitivity. The aim of the present study was to assess the effects of a high-fat diet (HFD), fed at two energetic levels, on microbiota, intestinal barrier, and inflammatory and metabolic parameters in dogs. A HFD (33% fat as fed, 4,830 kcal/kg) was given to 24 healthy Beagle dogs at 100% (HF-100; n = 8) and at 150% (HF-150; n = 16) of their maintenance energy requirements for 8 weeks. Analysis of similarity revealed a significant difference in gut microbiota ß-diversity following the diet compared to week 0 in both groups while α-diversity was lower only in the HF-150 group. Firmicutes/Bacteroidetes ratio was higher in the HF-150 group compared to the HF-100 group at weeks 2 and 8. A reduction in insulin sensitivity was observed over time in the HF150 group. Neither endotoxemia nor inflammation was observed in either group, did not find supporting data for the hypothesis that the microbiota is involved in the decline of insulin sensitivity through metabolic endotoxemia and low-grade inflammation. Colonic permeability was increased at week 4 in both groups and returned to initial levels at week 8, and was associated with modifications to the expression of genes involved in colonic barrier function. The increase in intestinal permeability may have been caused by the altered intestinal microbiota and increased expression of genes encoding tight junction proteins might indicate a compensatory mechanism to restore normal permeability. Although simultaneous changes to the microbiota, barrier permeability, inflammatory, and metabolic status have not been observed, such a causal link cannot be excluded in dogs overfed on a HFD. Further studies are necessary to better understand the link between HFD, intestinal microbiota and the host.

Effect of dietary polyunsaturated fatty acids on the expression of peroxisomal ABC transporters.

Peroxisomal ABC transporters encoded by the ABCD genes are thought to participate in the import of specific fatty acids in the peroxisomal matrix. ABCD1 deficiency is associated with X-linked adrenoleukodystrophy (X-ALD), the most frequent peroxisomal disorder which is characterized by the accumulation of saturated very-long-chain fatty acids (VLCFA). ABCD2 (the closest homolog of ABCD1) and ABCD3 have been shown to have partial functional redundancy with ABCD1; only when overexpressed, they can compensate for VLCFA accumulation. Other lipids, for instance polyunsaturated fatty acids (PUFA), should be possible candidate substrates for the ABCD2 and ABCD3 gene products, ALDRP and PMP70 respectively. Moreover, PUFA, which are known regulators of gene expression, could therefore represent potent inducers of the ABCD genes. To test this hypothesis, littermates of n-3-deficient rats were subjected to an n-3-deficient diet or equilibrated diets containing ALA (alpha-linolenic acid, 18:3n-3) as unique source of n-3 fatty acids or ALA plus DHA (docosahexaenoic acid, 22:6n-3) at two different doses. We analyzed the expression of peroxisomal ABC transporters and of the peroxisomal acyl-CoA oxidase gene 1 (Acox1) in adrenals, brain and liver. Whatever the diet, we did not observe any difference in gene expression in adrenals and brain. However, the hepatic expression level of Abcd2 and Abcd3 genes was found to be significantly higher in the n-3-deficient rats than in the rats fed the ALA diet or the DHA supplemented diets. This was accompanied by important changes in hepatic fatty acid composition. In summary, the hepatic expression of Abcd2 and Abcd3 but not of Abcd1 and Abcd4 appears to be highly sensitive towards dietary PUFA. This difference could be linked to the substrate specificity of the peroxisomal ABC transporters and a specific involvement of Abcd2 and Abcd3 in PUFA metabolism.

The hyperenergetic-fed obese dog, a model of disturbance of apolipoprotein B-100 metabolism associated with insulin resistance: kinetic study using stable isotopes.

The hyperenergetic-fed beagle dog model of obesity-associated insulin resistance has previously demonstrated lipoprotein abnormalities similar to those of obese insulin-resistant humans. The aim of this study was to check, in the insulin-resistant dog, the mechanism leading to abnormalities in the mass of apolipoprotein B-100 (apo B-100) containing lipoproteins. Six healthy male beagle dogs were overfed with a high-fat diet for 28 +/- 2.5 weeks. Obesity was associated with insulin resistance as assessed by the euglycemic hyperinsulinemic clamp technique. The kinetics of very low-density lipoprotein (VLDL) and low-density lipoprotein (LDL) apo B-100 were recorded in dogs, at healthy and insulin-resistant states, using a primed constant infusion of [5,5,5-D(3)]leucine. Isotopic enrichment was measured by gas chromatography-mass spectrometry (GC-MS). A multicompartmental model was used for the analysis of tracer kinetics data. Apolipoprotein B-100 concentration was higher in VLDL (2.8-fold, P < .05) but lower in LDL (2-fold, P < .05) in the insulin-resistant compared to the healthy state. Kinetic analysis showed a higher VLDL apo B-100 production (1.7-fold, P < .05). The fractional catabolic rate of VLDL did not change significantly, but the lipolysis was decreased significantly (3-fold, P < .05). The lower LDL apo B-100 level in insulin-resistant dogs was explained by a higher LDL fractional catabolic rate (2.5-fold, P < .05). The mechanisms leading to hypertriglyceridemia (higher production rate and lower lipolysis of VLDL) in insulin-resistant dogs were similar to those described in the insulin-resistant humans.

Use of water turnover method to measure mother's milk flow in a rat model: Application to dams receiving a low protein diet during gestation and lactation.

Assessment of milk production is of utmost relevance for pediatricians and scientists interested in early life nutrition. The weight-suckle-weight (WSW) method, which consists of weighing babies before and after they suckle their mother, uses the difference in body weight as an estimate of milk intake. However, this is prone to many sources of error. In the current study, we used for the first time the water turnover method and compartmental analysis with deuterated water (D2O) as a non-toxic tracer to quantify in vivo milk production in a rat model. We assessed the effect of a nutritional intervention presumed to affect milk production, a maternal dietary protein restriction during gestation and lactation, which results in the birth of pups with intrauterine growth restriction. The specific aim of this study was to determine milk production with the body water turnover method in rat dams receiving during gestation and lactation, either a control diet (NP) or an iso-caloric low-protein diet (LP). In NP dams, mass of dam's total body water, output flow constant from dam to litter (K21) and median milk flow, calculated between days 11 to 14 after pup birth, were 282.1 g, 0.0122 h-1 and 3.30 g/h for NP dams, respectively. Maternal dietary protein restriction (-59%) during perinatal period led to a 34% reduction in milk flow (NP versus LP). With the WSW method, milk flow varied from 1.96 g/h to 2.37 g/h between days 11 to 14 for NP dams. The main advantage of the D20 method compared to the WSW method stems from its higher precision, as attested by the narrowest range of measured values of milk flow ([2.90; 3.75] and [0.98; 6.85] g/h, respectively) for NP group. This method could be suitable for testing the effectiveness of candidate galactologue molecules presumed to enhance milk production in the lactating rat model.

In situ hybridization of dihydroxyacetone phosphate acyltransferase, the regulating enzyme involved in plasmalogen biosynthesis.

In situ hybridization can be carried out using different methods. The experimenter has to choose various parameters: the type of tissue fixation, the time of incubation, and the duration of the exposure time. All these parameters are determinant for the sensitivity and the resolution of this technique. This publication of technical aspects described different experiments performed for in situ hybridization on liver tissue. We may conclude on the parameters to optimize each step of the hybridization procedure. Moreover, this technique could be transposed to the brain and applied to little structures with a light expression of DHAP-AT.

Nanophotonic reservoir computing with photonic crystal cavities to generate periodic patterns.

Reservoir computing (RC) is a technique in machine learning inspired by neural systems. RC has been used successfully to solve complex problems such as signal classification and signal generation. These systems are mainly implemented in software, and thereby they are limited in speed and power efficiency. Several optical and optoelectronic implementations have been demonstrated, in which the system has signals with an amplitude and phase. It is proven that these enrich the dynamics of the system, which is beneficial for the performance. In this paper, we introduce a novel optical architecture based on nanophotonic crystal cavities. This allows us to integrate many neurons on one chip, which, compared with other photonic solutions, closest resembles a classical neural network. Furthermore, the components are passive, which simplifies the design and reduces the power consumption. To assess the performance of this network, we train a photonic network to generate periodic patterns, using an alternative online learning rule called first-order reduced and corrected error. For this, we first train a classical hyperbolic tangent reservoir, but then we vary some of the properties to incorporate typical aspects of a photonics reservoir, such as the use of continuous-time versus discrete-time signals and the use of complex-valued versus real-valued signals. Then, the nanophotonic reservoir is simulated and we explore the role of relevant parameters such as the topology, the phases between the resonators, the number of nodes that are biased and the delay between the resonators. It is important that these parameters are chosen such that no strong self-oscillations occur. Finally, our results show that for a signal generation task a complex-valued, continuous-time nanophotonic reservoir outperforms a classical (i.e., discrete-time, real-valued) leaky hyperbolic tangent reservoir (normalized root-mean-square errors=0.030 versus NRMSE=0.127).

CETP inhibitor torcetrapib promotes reverse cholesterol transport in obese insulin-resistant CETP-ApoB100 transgenic mice.

Insulin resistance and type 2 diabetes are associated with low HDL-cholesterol (HDL-c) levels, which would impair reverse cholesterol transport (RCT). A promising therapeutic strategy is to raise HDL with cholesteryl ester transfer protein (CETP) inhibitors, but their effects on RCT remains to be demonstrated in vivo. We therefore evaluated the effects of CETP inhibitor torcetrapib in CETP-apolipoprotein (apo)B100 mice made obese and insulin resistant with a 60% high-fat diet. High-fat diet over 3 months increased body weight and homeostasis model of insulin resistance index by 30% and 846%, respectively (p < 0.01 for both vs. chow-fed mice). Total cholesterol (TC) increased by 46% and HDL-c/TC ratio decreased by 28% (both p < 0.05). Compared to vehicle, high-fat-fed mice treated with torcetrapib (30 mg/kg/day, 3 weeks) showed increased HDL-c levels and HDL-c/TC ratio by 41% and 37% (both p < 0.05). Torcetrapib increased in vitro macrophage cholesterol efflux by 22% and in vivo RCT through a 118% increase in (3) H-bile acids fecal excretion after (3) H-cholesterol labeled macrophage injection (p < 0.01 for both). Fecal total bile acids mass was also increased by 158% (p < 0.001). In conclusion, CETP inhibition by torcetrapib improves RCT in CETP-apoB100 mice. These results emphasize the potential of CETP inhibition to prevent cardiovascular diseases.

Plasmalogens in the retina: in situ hybridization of dihydroxyacetone phosphate acyltransferase (DHAP-AT)--the first enzyme involved in their biosynthesis--and comparative study of retinal and retinal pigment epithelial lipid composition.

Plasmalogens (Pls) are phospholipids containing a vinyl-ether bond in the sn-1 position of the glycerol backbone. The physiological role of Pls is still enigmatic, especially within the eye where their deficiency leads to developmental abnormalities. In order to learn more about the functions of Pls in the posterior eye, we evaluated retinal Pl content as well as the expression of the first enzyme involved in Pls biosynthesis, dihydroxyacetone phosphate acyltransferase (DHAP-AT) in the retina. In situ hybridization of DHAP-AT mRNA was performed on rat eye sections. The Pl contents of calf retina and retinal pigment epithelium (RPE) samples were determined by high-performance liquid chromatography, thin-layer chromatography, and gas chromatography. DHAP-AT was highly expressed in the inner segment of photoreceptors and in the RPE, suggesting two distinct sites for Pl biosynthesis. Plasmenyl-ethanolamine was the prominent class of Pls in both neural retina and RPE (28-29% of the total phospho-ethanolamine-glycerides). According to the nature of the alkenyl residue linked to the sn-1 position of Pls, the most striking finding was the greater proportion of octadecanal-aldehyde in the sn-1 position of plasmenyl-ethanolamine of the neural retina compared to all the other classes of Pls in the neural retina and the RPE. These findings might be relevant to the biological functions of Pls against oxidative stress and in the formation of lipid rafts.

Dietary oxysterols induce in vivo toxicity of coronary endothelial and smooth muscle cells.

Dietary cholesterol oxidation products (COPs) were reported to exhibit in vitro toxicity toward vascular cells. The aim of this study was to determine whether dietary COPs induce in vivo toxicity toward coronary arteries and to evaluate their effect on the coronary reactivity. Golden Syrian hamsters were fed either a normolipidic diet or a hyperlipidic diet with or without a mixture of COPs (1.4 mg/kg/day). At the end of the feeding periods, cardiac mitochondria and cytosol were prepared to determine the subcellular distribution of cytochrome c. Oxidative phosphorylation was evaluated with glutamate, pyruvate or palmitoylcarnitine as a substrate. The main coronary artery was examined all along its length by transmission electron microscopy (TEM). Plasma sterol concentrations were determined. Furthermore, at the end of the 3-month feeding period, the hearts were perfused at constant pressure by the Langendorff method. The endothelium-dependent reactivity to acetylcholine was evaluated. The myocardial sterol concentration was also estimated. After a 15-day diet with dietary COPs, a release of cytochrome c into the cytosolic fraction of the whole heart occurred, which indicated apoptosis of one or several types of cardiac cells probably induced by excess circulating cholestanetriol. The morphological data obtained by TEM after three months of diet suggested that mainly vascular cells (endothelial and smooth muscle cells) were damaged by dietary COPs, whereas cardiomyocytes appeared healthy. Furthermore, the mitochondrial oxidation of palmitoylcarnitine was reduced and that of pyruvate was increased, suggesting some maintenance of energy metabolism. This strengthens the hypothesis of apoptosis. Several changes in coronary reactivity suggesting an increased NO production were observed. In conclusion, dietary COPs triggered in vivo apoptosis of coronary cells through the release of cytochrome c in the cytosol. This toxicity was counterbalanced by an increased endothelium-dependent dilation.

Fatty acid oxidation and related gene expression in heart depleted of carnitine by mildronate treatment in the rat.

The metabolic and genic effects induced by a 20-fold lowering of carnitine content in the heart were studied in mildronate-treated rats. In the perfused heart, the proportion of palmitate taken up then oxidized was 5-10% lower, while the triacylglycerol (TAG) formation was 100% greater than in controls. The treatment was shown to increase the maximal capacity of heart homogenates to oxidize palmitate, the mRNA level of carnitine palmitoyltransferase I (CPT-I) isoforms, the specific activity of CPT-I in subsarcolemmal mitochondria and the total carnitine content of isolated mitochondria. Concomitantly, the increased mRNA expression of lipoprotein lipase, fatty acid translocase and enzymes of TAG synthesis was associated with a 5- and 2-times increase in serum TAG and free fatty acid contents, respectively. The compartmentation of carnitine at its main functional location was expected to allow the increased CPT-I activity to ensure in vivo correct fatty acid oxidation rates. All the inductions related to fatty acid transport, oxidation and esterification most likely stem from the abundance of blood lipids providing cardiomyocytes with more fatty acids.