Dunedin, NZ) that gave an output of renal blood flow in litres per minute. To avoid vasoconstriction (as a re- sult of manipulation) confounding our results, a 10- minute rest or nonmanipulation period was provided prior to baseline measurements. This was followed by a 5- minute period of baseline flow measurements and then infusion of acetylcholine or sodium nitroprusside for 5 mi- nutes during which flow was continuously monitored. The mean flow during the time period was calculated and flow measurements were performed in triplicate both with and without acetylcholine/sodium nitroprusside. Endothe- lial dysfunction was determined by the change in renal blood flow in response to an acetylcholine infusion (0.1 to 10 μg/kg/minute) administered via a 14G peripheral ven- ous cannula (BD Venflon, Becton Dickinson, Oxford, UK) inserted into the suprarenal abdominal aorta. Sodium ni- troprusside (0.1 to 10 nmol/kg/minute) was used to con- trol for endothelium-independent relaxation. Renal cortical and medullary nucleotide levels were measured using reverse-phase high-performance liquid chromatog- raphy as described previously [17,18].
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Activation of central PPARγ promotes food intake and body weight gain; however, the identity of the neurons that express PPARγ and mediate the effect of this nuclear receptor on energy homeostasis is unknown. Here, we determined that selective ablation of PPARγ in murine proopiomelanocortin (POMC) neurons decreases peroxisome density, elevates reactive oxygen species, and induces leptin sensitivity in these neurons. Furthermore, ablation of PPARγ in POMC neurons preserved the interaction between mitochondria and the endoplasmic reticulum, which is dysregulated by HFD. Compared with control animals, mice lacking PPARγ in POMC neurons had increased energy expenditure and locomotor activity; reduced body weight, fat mass, and food intake; and improved glucose metabolism when exposed to high-fat diet (HFD). Finally, peripheral administration of either a PPARγ activator or inhibitor failed to affect food intake of mice with POMC-specific PPARγ ablation. Taken together, our data indicate that PPARγ mediates cellular, biological, and functional adaptations of POMC neurons to HFD, thereby regulating whole-body energy balance.
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Apolipoprotein E serves as a TRL ligand for both the LDL receptor and LRP [24,25]. The liver is a major source of apo E, however other tissues including the small intestine express apo E [24,26,27]. Apo E KO mice [28-30] accumu- late TRL's because they are unable to bind and be cleared by receptor processes . Under low-fat feeding, apo E KO mice had a greater than three-fold increase in plasma. High-fat feeding exacerbated plasma cholesterol accumu- lation in apo E KO mice, presumably because of exagger- ated lipoprotein production and indeed hypercholesterolemia was increased two-fold above low- fat fed apo E KO mice. Clearance of TRL's from blood is a two-step process requiring triglyceride lipolysis by lipases to produce a depleted apo E rich 'remnant' lipoprotein . Thereafter, remnants are cleared by receptor path- ways utilizing apo E as the ligand. There is no hydrolytic defect in apo E KO mice, which explains why these mice were not hypertriglyceridemic.
It was apparent from the body-weight proﬁles of the ani- mals that maternal diet, maternal age and high-fat feeding all made an impact on adiposity. The resistance to fat deposition in LP-exposed animals was not fully explained by lower energy intake, so in order to determine whether differences in fat gain were explained by altered energy expenditure through physical activity, locomotor activity was determined in all animals at half way through the high-fat feeding trial. As shown in Fig. 1(c) and (d), the rearing of female offspring was not sig- niﬁcantly inﬂuenced by maternal age, maternal diet or high-fat feeding. Similarly the mobility of females was not inﬂuenced by any of the factors under study (Fig. 2(c) and (d)), but activ- ity was signiﬁcantly increased by feeding the high-fat diet (Fig. 3(c) and (d)). Among the males, offspring of older mothers tended to rear more than those from younger mothers, whilst those exposed to the LP diet in utero were slightly less exploratory (Fig. 1(a)). Consumption of the high- fat diet also had a small but signiﬁcant effect on rearing (Fig. 1(b)). Male offspring born to older mothers were observed to be less mobile in a novel environment (Fig. 2(a) and (b)), but this behaviour was not inﬂuenced by maternal or high-fat diet. The activity of male offspring was not inﬂu- enced by maternal factors or the high-fat diet (Fig. 3(a) and (b). Thus variation in locomotor activity did not appear to explain differences in weight gain or adiposity. The difference in rearing activity was, however, an interesting behaviour for further investigation.
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To normalize expression data, we used multiple internal control genes as described by Vandesompele et al. (49). The internal control genes encoding HMG-14 (Hmg14) and ribosomal protein S3 (Rps3) were selected from our murine microarray data set for having high expres- sion and little sample-to-sample variability. For each transcript assayed, intron-spanning primers were designed using publicly available genomic contig sequences obtained through Entrez Gene (http://www.ncbi.nlm. nih.gov/entrez/query.fcgi?db=gene), the public domain primer design software Primer3 (http://frodo.wi.mit.edu/primer3/primer3_code.html), and the DNA analysis software Vector NTI Suite Version 7 (Informax Inc.). Primer sequences were as follows: Rps3 forward, 5′-ATCAGAGA- GTTGACCGCAGTTG-3′; Rps3 reverse, 5′-AATGAACCGAAGCACAC- CATAG-3′; Emr1 forward, 5′-CTTTGGCTATGGGCTTCCAGTC-3′; Emr1 reverse, 5′-GCAAGGAGGACAGAGTTTATCGTG-3′; Cd68 for- ward, 5′-CTTCCCACAGGCAGCACAG-3′; Cd68 reverse, 5′-AATGAT- GAGAGGCAGCAAGAGG-3′; Tnfa forward, 5′-CCAGACCCTCACTA- GATCA-3′; Tnfa reverse, 5′-CACTTGGTGGTTTGCTACGAC-3′; Acdc forward, 5′-GCTCCTGCTTTGGTCCCTCCAC-3′; Acdc reverse, 5′- GCCCTTCAGCTCCTGTCATTCC-3′; Hmg14 forward, 5′-GCAGAAAAT- GGAGAGACGGAAAACC-3′; Hmg14 reverse, 5′-AAGGGAGGCGGGAC- CACTGAC-3′; Ccl2 forward, 5′-AGGTCCCTGTCATGCTTCTGG-3′; Ccl2 reverse, 5′-CTGCTGCTGGTGATCCTCTTG-3′; Pparg forward, 5′- GCCCTTTGGTGACTTTATGGAG-3′; Pparg reverse, 5′-GCAGCAGGTT- GTCTTGGATG-3′; Lipe forward, 5′-ACGAGCCCTACCTCAAGAACTG- 3′; Lipe reverse, 5′-ATCTGGCACCCTCACTCCATAG-3′; Fabp4 forward, 5′-AAGAAGTGGGAGTGGGCTTTG-3′; Fabp4 reverse, 5′-CTGTC- GTCTGCGGTGATTTC-3′; Serpine1 forward, 5′-TCCTCATCCTGCCTA- AGTTCTC-3′; Serpine1 reverse, 5′-GTGCCGCTCTCGTTTACCTC-3′; Gpam forward, 5′-TCCAGAAGGTGAAAAGGAAAGC-3′; Gpam reverse, 5′-GGCAAAAGAGGATGAAGGTGAG-3′.
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The results of our experiments show that plasma Hcy is significantly increased in high fat diet-induced NAFLD in rats, and that this change is accompanied by a decrease in plasma Cys (Figure 2A,B), suggesting that the transsulphuration pathway is affected. Determination of the activities of the two enzymes of the pathway showed that both are significantly down-regulated in high fat diet-induced NAFLD (Figure 2C,D). We found no evidence, however, for changes in the expression of other key enzymes in the hepatic methionine cycle, including MAT1A, the gene which encodes the catalytic subunit of the isoenzymes MATI and MATIII which are expressed in adult liver , and the methyltransferases GNMT and PEMT (Table 3). Since these enzymes are methyltransferases while CBS and CGL are lyases, it is likely that they are regulated by different mechanisms. In addition, we have only measured mRNA expression for these enzymes, thus we cannot rule out the possibi- lity that there may be post transcriptional changes which modulate their activity in response to high fat feeding. In contrast to our results, Kwon et al.  have reported that CBS activity was unchanged in rats in which NAFLD was induced by feeding a diet containing 71% of the energy as fat, while CGL activity was increased by about 35%. In this study, however, the con- trol diet contained 35% of energy from fat, which is con- siderably higher than the 10% of energy from fat in our control diet. Thus, our findings indicate that high fat diet-induced NAFLD is associated with HHcy, and that this is caused by reduced conversion to Cys via the transsulphuration pathway, while the expression of methyltransferases involved in liver methionine metabo- lism is not changed. As it has been estimated that as much as 50% of the Cys required for GSH synthesis is formed from Hcy via this route, and the availability of Cys is a limiting factor for GSH production [12,31], in the long term this may lead to a decrease in body levels of this antioxidant.
Beta2-integrins are important in leukocyte trafficking and function, and are regulated through the binding of cytoplasmic proteins, such as kindlin-3, to their intracellular domain. Here, we investigate the involvement of beta2-integrins in the regulation of metabolic dis- ease using mice where the kindlin-3 binding site in the beta2-integrin cytoplasmic tail has been mutated (TTT/AAA-beta2-integrin knock-in (KI) mice), leading to expressed but dys- functional beta2-integrins and significant neutrophilia in vivo . Beta2-integrin KI mice fed on a high fat diet showed normal weight gain, and normal accumulation of macrophages and lymphocytes in white adipose tissue (WAT) and liver, but increased neutrophil numbers especially in WAT. In addition, beta2-integrin KI mice fed on a high fat diet showed signifi- cantly increased peripheral insulin resistance in response to high-fat feeding. However, this was associated with improved glucose disposal following glucose load. Interestingly, beta2- integrin KI neutrophils produced more elastase in vitro , in response to stimulation. Beta2- integrin KI mice displayed variability of tissue inflammatory status, with liver and WAT exhib- iting little or no difference in inflammation compared to high fat fed controls, whereas skele- tal muscle demonstrated a raised inflammatory profile in association with higher elastase levels and diminished signalling through the IRS1-PKB pathway. In conclusion, although expression of dysfunctional beta2-integrins increased neutrophil production and infiltration into tissue, skeletal muscle was the most affected tissue exhibiting evidence of higher neu- trophil activity and insulin resistance. Thus, beta2-integrins modulate glucose homeostasis during high fat feeding predominantly through actions on skeletal muscle to affect metabolic phenotype in vivo .
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Exercise training concomitant to high-fat diet feeding reduces degree of insulin resistance and improves adipoR1/APPL1 protein levels in the adipose tissue The effects of exercise training on levels of phosphory- lation of IR, IRS1, and Akt were examined in the adipose tissue of C, C-T, DIO, and DIO-T groups submitted to the exercise training. As expected, an insulin (+) injec- tion showed increased IR, IRS, and Akt phosphorylation (Figures 1A, 1B, and 1C, respectively) in the adipose tis- sue of mice that were fed chow standard (sedentary and training groups) when compared with those that were administered a saline (−) injection. However, when mice were fed with a high-fat diet and not submitted to train- ing protocol (DIO group), IR, IRS, and Akt phosphory- lation (Figures 1A, 1B, and 1C, respectively) in adipose tissue were reduced, when compared with control group. On the other hand, IR, IRS, and Akt phosphorylation (Figure 1A, B, and C, respectively) in the adipose tissue of the DIO + T group increased, when compared with the DIO sedentary group. In parallel, the adiponectin re- ceptor 1 (ADIPOR1) and its molecule adaptor APPL1 exhibit market levels reduced when compared with control sedentary and training groups (Figure 1D and 1E, respectively). However, when exercise training was
der and obesity appeared in more and more people, also, the function of the male reproduc- tive system had a certain negative affect by the abnormal lipid metabolism. Researches showed that people with obesity disorders, par- ticularly male, had reproductive function re- duced and testosterone levels decreased . We found The level of serum testosterone was significantly lower in the group of abnormal lipid metabolism rats fed with high fat diet com- pared with the control group, which illustrated lipid metabolism inhibited the synthesis of tes- tosterone. The western blot analysis showed that the expression levels of StAR and P450 scc of rat leydig cells in the group of abnormal Figure 2. The expression levels of StAR and P450 scc of rat leydig cells by
The content of chosen components in cows’ co- lostrum is showed in Tab. III. There were no statis- tically significant differences between the groups. The highest content of dry matter (263.2 g/l) and lac- tose (19.13 g/l) was noted in colostrum of cows from group III, however, the content of fat (51.28 g/l) was the highest in group II. The increased fat content in colostrum in this group could possibly be explained by the formation of so-called chylomicrons. They are synthesised from precursors of glucose and af- ter their release into milk they increase the amount of detectable milk fat (Jelínek and Koudela, 2003). The smallest content of α-lactoglobulines and β-lactoglo- bulines was stated in cows’ colostrum from group II. The largest content of β-lactoglobulines was found in group III (109.59 g/l). The content of colostrum com- ponents for all the cows was similar to the results ob- tained by others authors (Zachwieja, 1991 and 1995; Blum and Hammon, 2000). Taking of colostrum by the calves directly after the birth in the right amount is very important with respect to gaining the immunity and because of the content of hydroxycarbons, lipids, proteins, mineral elements and vitamins (Zachwieja, 1991; Blum and Hammon, 2000). Despite of the lack of statistic differences, the more profitable composi- tion of colostrum was stated in group III receiving the highest dose of propylene glycol. There are no works concerning the influence of propylene glycol used in cows’ feeding before parturition onto colostrum com- position, thus it is difficult to compare the results.
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The animals were either fed normal fat diet (NFD) or high fat diet (HFD). The NFD comprised of standard rat chow (Amrut Laboratory Animal feed, Maharashtra, India) containing protein 22.10%, oil 4.13%, fibre 3.15%, ash 5.15%, sand (silica) 1.12% w/w), whereas, HFD was composed of standard rat chow - 68%, Dalda (saturated fat) - 30% and cholesterol - 2% (14).
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Our data shows xylitol mice experienced the highest weight reduction compared to other groups. There are many different factors involved in weight control. One of it is the gastric emptying time. The slower gastric emptying time for the digestion and absorption of nutrients affects the body weight loss. Previous studies have reported that the consumption of xylitol had significantly prolonged the gastric emptying time  . It not only prolongs gastric emptying time but also concomitantly accelerates the intestinal transit of nutrients compared to glucose when fed as a single oral dose . The parenteral infusion of glucose substitutes including xylitol has been reported to have several health benefits, such as anti-ketogenic effect, small damaging effect towards vein as well as increased in metabolism . Compared to glucose, xylitol consumption is more rapidly followed by high glycogen storage in the liver . Later this will reduce the generation of glucose from amino acids through gluconeogenesis . Improved nitrogen balanced, augmented level of protein and muscle RNA content were found in critically ill animal infusion by xylitol  .
Given the common and early presence of endothelial dysfunc- tion in dyslipidemia, obesity, and diabetes and PPARγ as a regula- tor of energy balance expressed in the endothelium, we hypoth- esized that endothelial PPARγ might be involved in directing metabolic phenotype. To test this, we studied mice deficient in endothelial PPARγ, using floxed PPARγ mice and a Cre transgene under control by the endothelial/hematopoietic specific Tie2 pro- moter (11). We investigated these mice under conditions of stan- dard chow and high-fat diet, both with and without treatment with the PPARγ agonist rosiglitazone as well as before and after BM transplantation (BMT) to reconstitute hematopoietic PPARγ expression and isolate endothelial PPARγ-dependent responses. These studies reveal that mice specifically deficient in endothelial PPARγ manifest a distinct pattern of decreased adiposity, increased insulin action, worsened dyslipidemia, and impaired arterial vaso- dilation in response to high-fat diet challenges as compared with control mice. Moreover, these mice fail to exhibit known meta- bolic improvements in response to a PPARγ agonist. These data establish PPARγ in the endothelium as a previously unrecognized determinant of metabolic status and a potential contributor to metabolic abnormalities found in insulin resistance and diabetes. Results
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expected increase in levels as would be expected from feeding the mice a HF-diet. The ipGTT insulin levels of the chow and HF-fed B10 k mice were very similar to those of the respective Balb/c mice, but this was unexpected considering the hyperglycaemic glucose tolerance curves of both the chow and HF-fed B10 k mice. Furthermore, the chow-fed B10 k mice did not show the normal increase in insulin that is expected at the 15 min time point. Thus, a failure of insulin secretion to respond to hyperglycaemia is likely to be a factor contributing to the severely abnormal glucose tolerance of B10 k mice. Again, in marked contrast, the insulin levels of the NOD k mice were profoundly increased in the HF-fed compared to the chow-fed NOD k mice. Insulin secretion in the chow-fed NOD k mice appeared to be normal at which time glucose tolerance was normal. Fasting levels of insulin, however, increased by about 9 fold in response to them being on HF-diet. Furthermore, the pattern of secretion was very abnormal in the HF-fed NOD k , which showed a fall rather than increase in levels at 15 min compared to the fasting levels. This is indicative of the islet -cells of the NOD k mice at least trying
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In the present work, serum cholesterol and LDL cholesterol concentrations were not modified by the HF- CLAb diet compared to the NF-So and HF-Cb diets, re- spectively. Similarly, no effects of cis-9, trans-11 CLA on cholesterol and LDL cholesterol levels were also shown previously [36,37]. The high LDL cholesterol concentra- tion in NF-So-fed rats may be due to high levels of carbohydrate (73.39% of energy) in this diet, since it was demonstrated that when dietary carbohydrate was in- creased from 50% to 67% of energy, the fasting triacyl- glycerol level rose , which is commonly related to increased precursors of LDL cholesterol in the blood, the very-low-density lipoproteins, and consequently increased LDL cholesterol levels . Decreased total cholesterol concentration in HF-Cb or HF-So-fed rats was related to the low HDL cholesterol level in these groups, which is a risk factor for type 2 diabetes mellitus .
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production, we defined a butyrate formation module in- cluding enzymes catalyzing 8 reactions from acetyl-CoA to butyrate (Fig. 5b). The relative abundance of the 11 KOs comprising this module was calculated according to the different subgroups. In line with the results shown in Fig. 5b, the relative abundance of KOs comprising this butyrate formation module was higher in Sv129 mice than in BL6 mice (Additional file 14: Figure S12). This suggests that differences in butyrate production of the gut microbiota in Sv129 and BL6 mice might contribute to the different phenotypes in relation to obesity propen- sity. We used a previously reported z score method  to further analyze diet-dependent differences between the two strains at the functional level. Main differences are listed in Additional file 15: Table S3. These analyses indicated that the metabolism of short chain fatty acids (SCFAs) was differently affected by the HF diet in the two strains. In keeping with results on the potential for butyrate production, HF feeding was associated with a general increase in the abundance of genes involved in butyrate metabolism in both strains (Additional file 14: Figure S12), whereas the abundance of genes involved in propionate metabolism module, associated with in- creased energy harvest, increased in BL6 mice in re- sponse to HF feeding (Additional file 15: Table S3), possibly at least in part contributing to the sensitivity of the BL6 strain to HF diets. Still, in relation to obesity propensity, more experiments are clearly needed to ex- plain the increase in the abundance of BCT in response to HF feeding.
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weight and other proxies of adiposity do not accurately capture body fat content. 23 To better understand how nutrition modulates early patterns of adipose tissue deposition, numerous studies and meta-analyses have also investigated body composition differences among breastfed and formula-fed infants over the first year of life with methods ranging from anthropometry to a wide variety of in-vivo measurement techniques. 10,20,50-53 These studies have also employed a variety of outcome measures, the majority of which have been total body fat or percentage body fat. While these studies have not assessed the direct effect of diet on adipose tissue cellularity, a reported positive association between cell size, cell number, and percent body fat 31 provides insight into how these studies may translate at the cellular level. These studies do not, however, describe how feeding practices affect the distribution of adipose tissue deposition. As it is critical to assess both the amount and distribution of adipose tissue, literature describing site-specific differences in adiposity is more limited.
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For the entre´e choice, the feeding goal block signifi- cantly associated with participants’ entre´e selection, be- yond that of the covariates (see Table 5). Feeding for natural content was significantly associated with choosing lower-calorie/-fat entre´es, whereas food familiarity was significantly associated with choosing higher-calorie/-fat entre´es. Approximately 25% of the variance was explained by the covariates and maternal feeding goals. Fast food frequency and maternal feeding goals accurately predicted approximately 70% of the entre´es chosen. This model had a higher sensitivity than specificity, with 78.6% correct prediction of choosing the higher-calorie/-fat options ver- sus 57.7% correct prediction of lower-calorie/-fat entre´e choices.
Eight adult mongrel dogs (5 males and 3 females) were trained to stand quietly in a padded sling. All dogs were then surgically instrumented with an ascending aortic catheter and 2 right atrial catheters. Surgical instrumentation of the animals was per- formed under methohexital induction (12 mg ⁄ kg) and isoflurane (0.5–1.5%). After surgery, the dogs were allowed to recover for 3 weeks before baseline measurements were made. Dogs received a regular diet (a diet that maintained weight constant), 1 to 2 cans of dog food (Ken-L-Ration; H. J. Heinz Com- pany, Pittsburgh, PA), for 2 weeks followed by 6 weeks of a high-fat diet consisting of approxi- mately 0.8 kg of cooked beef fat in addition to their regular diet. 17
Obesity and related disorders are becoming worldwide health issues [1 – 3]. Obesity is regarded as an inflamma- tory condition because of the associated low-grade in- flammation [4 – 6] affecting the periphery and increasing the incidence of many pathologies such as cardiovascular diseases , asthma , or even cancer . One of the proposed mechanisms leading to peripheral inflamma- tion implicates the gut microbiota. More specifically, a high-fat diet (HFD) will change the balance between dif- ferent populations of bacteria within the gut [10, 11]. This will lead to a disruption of the intestinal epithelium integrity that in turn will result in in- creased passage of endotoxins (such as lipopolysac- charides (LPS)) into the bloodstream that will then fuel the peripheral inflammatory tone [4, 12, 13]. The demonstration that disrupting LPS signaling (i.e., TLR4 − / − mice or CD14 − / − mice) protects from diet- induced obesity and metabolic disorders strongly sup- ports the important role played by LPS in the patho- physiology of these disorders [12 – 15].
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