Threonine utilization is high in the intestine of piglets

The whole-body threonine requirement in parenterally fed piglets is substantially lower than that in enterally fed piglets, indicating that enteral nutrition induces intestinal processes in demand of threonine. We hypothesized that the percentage of threonine utilization for oxidation and intestinal protein synthesis by the portal-drained viscera (PDV) increases when dietary protein intake is reduced. Piglets (n = 18) received isocaloric normal or protein-restricted diets. After 7 h of enteral feeding, total threonine utilization, incorporation into intestinal tissue, and oxidation by the PDV, were determined with stable isotope methodology [U-(13)C threonine infusion]. Although the absolute amount of systemic and dietary threonine utilized by the PDV was reduced in protein-restricted piglets, the percentage of dietary threonine intake utilized by the PDV did not differ between groups (normal protein 91% vs. low protein 85%). The incorporation of dietary threonine into the proximal jejunum was significantly different compared with the other intestinal segments. Dietary, rather than systemic threonine was preferentially utilized for protein synthesis in the small intestinal mucosa in piglets that consumed the normal protein diet (P < 0.05). Threonine oxidation by the PDV was limited during normal protein feeding. In protein-restricted pigs, half of the total whole-body oxidation occurred in the PDV. We conclude that, in vivo, the PDV have a high obligatory visceral requirement for threonine. The high rate of intestinal threonine utilization is due mainly to incorporation into mucosal proteins.     

Weaning affects the expression of heat shock proteins in different regions of the gastrointestinal tract of piglets

Heat shock proteins (HSP) play a central role in the protection of cells, tissues or organs subjected to various types of stressors. Different nutrients have been recently shown to exert their protection through the induction of HSP. Because these nutrients alleviate alterations of the intestine after weaning in pigs, this study was designed to obtain basic information on the expression of HSP 27, heat shock cognate 70 (HSC 70), HSP 70 and HSP 90 along the gastrointestinal tract (GIT) of young pigs and to study the effect of weaning on this expression. Pigs were weaned at 28 or 21 d and slaughtered at various times postweaning. All HSP were expressed in the GIT segments studied before and after weaning. However, the expression of HSP 27 and HSP 70 was transiently increased in the stomach and duodenum between 6 and 12 h postweaning and between 24 and 48 h in the mid-jejunum, ileum and colon. Their expressions were transiently decreased in the ileum. Expression of HSP 90 increased in the stomach and jejunum but decreased in the duodenum, ileum and colon. Similar results were obtained at both ages of weaning. We conclude that the HSP studied are present all along the gut of pigs and that their expression is modulated through weaning according to spatial-temporal patterns. The modulation by nutrients of HSP and their protective role on the GIT remain to be investigated in pigs.     

Dietary supplementation with zinc oxide increases Igf-I and Igf-I receptor gene expression in the small intestine of weanling piglets

This study was conducted to investigate the mechanism for the effect of elevated levels of dietary zinc oxide (ZnO) in enhancing the intestinal growth of weanling piglets. In Experiment 1, 4-wk-old (8.1 +/- 0.6 kg) crossbred barrows (n = 36) were assigned randomly to 1 of the 2 dietary groups, with 6 pens/group (3 pigs/pen). One group was fed the basal diet containing 100 mg Zn/kg diet. The other group was fed the basal diet supplemented with ZnO to provide 3000 mg Zn/kg diet. Pigs consumed their feed ad libitum for 14 d. In Experiment 2, 4-wk-old (7.6 +/- 0.16 kg) crossbred barrows (n = 16) were housed individually and assigned to 1 of the 2 dietary groups (8 pigs/group) as in Experiment 1, except that the 2 groups were pair-fed the same amount of feed. At the end of a 14-d treatment period, all of the pigs in both Experiments 1 and 2 were weighed, feed consumption was measured, and blood samples were collected for assays of insulin-like growth factor-I (IGF-I). In addition, 1 pig from each pen in Experiments 1 and 2 was selected randomly to obtain the small-intestinal mucosa for analyzing IGF-I and IGF-I receptor (IGF-IR) gene expression and to determine the small-intestinal morphology. In Experiment 1, dietary supplementation of ZnO increased (P < 0.05) the daily body weight gain and daily feed intake. In Experiment 2, dietary supplementation of ZnO increased (P < 0.05) the daily body weight gain and feed conversion efficiency. In both experiments, the villous height of the small-intestinal mucosa and both the mRNA and protein levels for IGF-I and IGF-IR in the small intestine were markedly enhanced (P < 0.05) by feeding elevated levels of Zn. Serum IGF-I levels did not differ between the control and Zn-supplemental groups in either experiment. Collectively, these results suggest that dietary Zn supplementation exerts its beneficial effects on the intestinal growth of weanling piglets through increasing IGF-I and IGF-IR expression in the small-intestinal mucosa.     

BBI抑制LPS诱导的肠道上皮细胞炎性因子的表达

目的探讨大豆来源的胰蛋白酶抑制剂(BBI)对LPS诱导的肠道上皮细胞炎性因子表达的抑制作用及其机制。方法用MTT法检测LPS和BBI对肠道上皮细胞的毒性作用。先用BBI预处理肠道上皮细胞,再用LPS刺激,采用实时荧光定量PCR检测细胞内炎性因子(TNF-α、IL-1β、IL-8和MCP-1)的表达,通过NF-κB-luc荧光素酶质粒和蛋白质印迹法(Western Blot)检测NF-κB的活性。结果 LPS最大浓度(10 000 ng/mL)和BBI最大浓度(1 000μg/mL)对细胞均无毒性作用。LPS处理可显著地上调肠道上皮细胞炎性因子(TNF-α、IL-1β、IL-8和MCP-1)的表达,其上调作用和LPS的浓度成正相关;LPS处理肠道上皮细胞3 h后炎性因子表达最高,随时间延长有所下降;LPS对肠道上皮细胞炎性因子的上调作用具有剂量和时间效应;BBI预处理肠道上皮细胞6 h时,BBI对LPS诱导肠道上皮细胞炎性因子表达的抑制作用最明显,且具有剂量效应。结论通过对肠道上皮细胞内NF-κB的抑制,BBI能够显著地抑制LPS诱导炎症因子的表达。     

Breastfeeding in infancy is not associated with inflammatory status in healthy adolescents

It has been suggested that breast-feeding (BF) may be associated with a decreased risk of cardiovascular disease in adulthood. A low-grade inflammation is associated with an increased risk of cardiovascular disease, even in apparently healthy children. The objective of this study was to assess the potential modulating effect of BF on the inflammatory status of healthy adolescents. Information on BF (duration) was obtained from parental records in 484 of 1040 healthy European urban adolescents (56.4% females) that had a blood sample obtained as part of the Healthy Lifestyle in Europe by Nutrition and Adolescence study. Blood serum inflammatory markers were measured, including high sensitivity C-reactive protein, complement factors 3 and 4, ceruloplasmin, adhesion molecules (L-selectin and soluble endothelial selectin, soluble vascular cell adhesion molecule 1, and intercellular adhesion molecule 1), cytokines, TGFβ1, and white blood cells. After univariate analysis, a propensity score, including the potential confounding factors, was computed and used to assess the association between BF and selected inflammatory markers. BF was not significantly associated with any of the selected inflammatory markers after adjustment for gender and propensity score. In our study, BF was not associated with low-grade inflammatory status in healthy adolescents, suggesting that the potential cardiovascular benefits of BF are related to other mechanisms than modulation of inflammation or might become relevant at a later age. Groups at high risk for cardiovascular disease should be a target for further research concerning the effects of BF.     

Cytokine-driven inflammatory response is associated with the hypermetabolism of AIDS patients with opportunistic infections

BACKGROUND: To assess a possible role of systemic inflammation in the resting metabolic response in AIDS patients with active secondary infections. METHODS: Fifty-two patients with AIDS-defined criteria and concomitant active infections and 19 healthy subjects were studied. Measurements were as follows: body composition assessed by bioelectrical impedance; resting energy expenditure (REE) by 30-minute indirect calorimetry; cytokine concentrations (IL-6, IFNalpha, TNFalpha, sTNF-R1) by ELISA; C-reactive protein (CRP), erythrocyte sedimentation rate, fibrinogen, and nutritional parameters by standard techniques. RESULTS: REE adjusted for fat-free mass (REEFFM) was significantly increased in AIDS patients despite 39% of them not being hypermetabolic. The patients were undernourished and were found to have increased levels of acute-phase proteins and increased concentrations of IL-6 and sTNF-R1 relative to controls. REE parameters were positively related to CRP, ESR, ferritin, IL-6, and sTNF-R1 and negatively related to albumin, prealbumin, and transferrin. CRP was an independent predictor of REEFFM in AIDS patients and explained 25% of its variability. Patients with severe inflammation (CRP > or = 37 mg/dL) were significantly hypermetabolic with respect to patients without inflammation (CRP < 6 mg/dL) and had higher levels of IL-6 and sTNF-R1 and lower levels of albumin and prealbumin. Although no significant differences were observed with respect to the infection type, patients with tuberculosis and Pneumocystis carinii infections had higher resting metabolic and inflammatory responses, whereas patients with recurrent bacterial pneumonia were normometabolic and had lower levels of inflammatory markers. CONCLUSIONS: Resting hypermetabolism observed in AIDS patients with concurrent active infections is related to the presence and severity of systemic cytokine-driven inflammatory response, which could reflect the type of secondary infection.     

Weaning anorexia may contribute to local inflammation in the piglet small intestine

Compromising alterations in villus-crypt structure are common in pigs postweaning. Possible contributions of local inflammatory reactions to villus-crypt alterations during the weaning transition have not been described. This study evaluated local inflammatory responses and their relationship with morphological changes in the intestine in 21-d-old pigs (n = 112) killed either at weaning (Day 0) or 0.5, 1, 2, 4 or 7 d after weaning to either milk- or soy-based pelleted diets. Cumulative intake averaged <100 g during the first 2 d postweaning, regardless of diet. During this period of weaning anorexia, inflammatory T-cell numbers and local expression of the matrix metalloproteinase stromelysin increased while jejunal villus height, crypt depth and major histocompatibility complex (MHC) class I RNA expression decreased. Upon resumption of feed intake by the fourth d postweaning, villus height and crypt depth, CD8(+) T cell numbers, MHC class I RNA expression and local expression of stromelysin returned to Day 0 values. Together the results indicate that inadequate feed intake during the immediate postweaning period may contribute to intestinal inflammation and thereby compromise villus-crypt structure and function.     

Hepatic abnormalities associated with aluminum loading in piglets

Cholestasis is a common complication of total parenteral nutrition (TPN) in infants. A contributing factor to the hepatic dysfunction may be a contaminant of the TPN solution, such as aluminum, that accumulates in liver and may act as a hepatotoxin. To study the hepatic effects of aluminum, growing piglets were given daily intravenous injections of aluminum, 1.5 mg/kg, for 50 days; pair-fed controls were given heparinized saline. At sacrifice, liver and serum were obtained. Liver was analyzed for histopathology and for aluminum content and localization. The hepatocyte lysosomes of the experimental group showed aluminum peaks by x-ray microanalysis, whereas the control group did not. No differences in ultrastructure were noted between the two groups when examined by electron microscopy. Mean serum total bile acid levels (27.8 +/- 15.9 SD vs 6.3 +/- 1.5 mumol/liter, p less than 0.05), mean alkaline phosphatase (309 +/- 108 vs 180 +/- 27 IU/liter, p = NS), and mean hepatic copper content (24.8 +/- 4.5 vs 14.4 +/- micrograms/g dry weight, p less than 0.01), were elevated in the aluminum-loaded piglets, indicating that cholestasis may have been produced. Also, a small but significant reduction in serum levels of 25 hydroxy-vitamin D was found in the aluminum-loaded piglets, suggesting that vitamin D hydroxylation may be impaired. Inasmuch as lysosomal contents are excreted into the bile, aluminum accumulation in lysosomes may alter lysosomal function and possibly affect bile flow or content.     

Increased colonic luminal synthesis of butyric acid is associated with lowered colonic cell proliferation in piglets

Butyrate inhibits colonic cell proliferation in vitro but reportedly has an opposite effect in vivo. Because lactulose feeding decreases cecal cell proliferation, an effect attenuated by prefeeding inulin, we hypothesized that lactulose feeding would decrease colonic luminal synthesis of butyrate, and that prefeeding and cofeeding inulin would prevent this effect. Piglets (n = 31) were catheterized and randomly assigned to 1 of 4 groups: Control formula (C); control formula + lactulose (L); control formula + lactulose + inulin (L + I); and control formula + inulin (I). At 6 and 7 d postsurgery, the rate of cecal synthesis of butyrate, cecal cell proliferation and apoptosis, and cecal and distal colon butyrate concentration were measured. In groups C, L, L + I, and I, the rates of synthesis of butyrate (mean +/- SEM) were 10.6 +/- 3.2, 23.3 +/- 4.5, 12.4 +/- 3.6, and 14.6 +/- 4.0 micromol/min, respectively (Group Effect, P = 0.1; C vs. L, P = 0.03; L vs. L + I, P = 0.06). The cecal butyrate concentrations did not differ among the 4 groups and were 8.7 +/- 3.2, 2.4 +/- 0.8, 3.4 +/- 1.9, and 2.0 +/- 0.7 micromol/g dry wt, respectively. The total cecal cell proliferation index was higher in C than in L (P = 0.008) or I (P = 0.026) and was higher in L + I than in L (P = 0.013) or I (P = 0.046). The increased supply of butyrate to the cecum was associated with decreased cell proliferation, but cecal butyrate concentration did not reflect synthesis.     

Onset of small intestinal atrophy is associated with reduced intestinal blood flow in TPN-fed neonatal piglets

Our aim was to determine the speed of onset of total parenteral nutrition (TPN)-induced mucosal atrophy, and whether this is associated with changes in intestinal blood flow and tissue metabolism in neonatal piglets. Piglets were implanted with jugular venous and duodenal catheters and either a portal venous or superior mesenteric artery (SMA) blood flow probe. At 3 wk of age, piglets were randomly assigned to receive continuous enteral formula feeding (n = 8) or TPN (n = 17) for 24 or 48 h. Blood flow was recorded continuously and piglets were given an i.v. bolus of bromodeoxyuridine and (13)C-phenylalanine to measure crypt cell proliferation and protein synthesis, respectively. After 8 h of TPN, portal and SMA blood flow decreased 30% compared with enteral feeding (P < 0.01), and remained near levels of food-deprived piglets for the remaining 48 h of TPN. After 24 h, TPN reduced jejunal inducible nitric oxide synthase (iNOS) activity and protein abundance (P < 0.05), small intestinal weight, and villous height (P < 0.01) compared with enterally fed piglets. Cell proliferation and DNA mass were decreased (P < 0.05) and apoptosis increased (P < 0.05) after 48 h of TPN. Protein synthesis was lower (P < 0.05) after 24 h of TPN, and protein mass was lower (P < 0.05) after 48 h of TPN, compared with enteral feeding. These data indicate that the transition from enteral to parenteral nutrition induced a rapid (<8 h) decrease in intestinal blood flow, and this likely precedes villous atrophy and the suppression of protein synthesis at 24 h, and of cell proliferation and survival at 48 h.     

A dried tofu-supplemented diet affects mRNA expression of inflammatory cytokines in human blood

In order to develop a new model of diet research, blood was drawn from 12 adult volunteers for 3 wk on regular diets as controls, and for a subsequent 3 wk supplemented with 18.5 g of freeze-dried tofu (Koya tofu) every day. Triplicate aliquots of 0.06 mL each of whole blood were stimulated ex vivo with phytohemagglutinin (PHA)-P, heat aggregated human IgG (HAG), lipopolysaccharide (LPS), zymosan A, and anti-T cell receptor (TCR) monoclonal antibody to activate specific subsets of leukocytes, then the levels of various inflammatory cytokine mRNA were quantified by real time PCR. Koya tofu significantly (p<0.05) augmented the fold increase of PHA-induced tumor necrosis factor superfamily (TNFSF) 15, IL6, and IL8, HAG-induced TNFSF15 and IL8, LPS-induced IL6 and IL8, zymosan-induced TNFSF15, IL6 and IL8, and TCR-induced TNFSF2 in comparison to the regular diet. Such increase was due to the reduction of baseline mRNA expression, not the enhancement of mRNA induction after specific stimulations. Six (TNFSF15), 4 (IL6), and 3 (IL10) subjects showed significant reduction of baseline mRNA during the Koya tofu diet compared to that of the control diet. Despite large individual-to-individual and day-to-day variation of mRNA, the method employed in this study was sensitive enough to identify statistically significant results as a group as well as on an individual basis, which will be a foundation for tailored diet in the future. The results also indicated that Koya tofu had a power to alter mRNA expression in leukocytes, and TNFSF15, IL6, and IL10 would be biomarkers for soy.     

慢性盆腔炎患者促炎因子与抗炎因子的关系

目的:探讨慢性盆腔炎患者血清促炎因子与抗炎因子的表达与相关性。方法:选择87例慢性盆腔炎患者作为病例组,选择同期健康体检妇女69例作为对照组,ELISA法检测血清TNF-α、IL-1β、IL-6和抗炎细胞因子IL-4、IL-10的表达。结果:病例组患者血清TNF-α、IL-1β和IL-6表达高于对照组(P<0.05),而血清IL-4和IL-10表达低于对照组(P<0.05);病例组患者血清促炎因子的表达与抗炎因子的表达呈负相关(P<0.05)。结论:慢性盆腔炎患者促炎因子过度激活,而抗炎因子被抑制,并且二者表达具有一定的相关性,共同促进慢性盆腔炎的发生发展。     

Exposure to welding fumes is associated with acute systemic inflammatory responses

Aims: To investigate the acute systemic inflammatory response to welding fume exposure.

Methods: Twenty four welders (42% smokers) and 13 non-exposed controls (23% smokers) were monitored at a welding school. Exposure to fine particulate matter (PM_2.5) was assessed using cyclone samplers. Markers of systemic inflammation, including C-reactive protein (CRP), fibrinogen, and white blood cell (WBC) levels, were determined in peripheral blood samples collected at baseline and after 5.3 (SD 1.0) hours of exposure.

Results: The median PM_2.5 concentration for welders was 1.66 mg/m³, which was significantly greater than that for controls (0.04 mg/m³). Compared to non-smokers, smokers had a significantly higher baseline WBC count, but comparable levels of CRP and fibrinogen. In non-smokers, welding fume exposure was associated with a significant increase in WBC and neutrophil counts immediately following exposure (+0.8x10³/µl, 95% CI 0.1 to 1.6, and +1.0x10³/µl, 95% CI 0.4 to 1.7, respectively). A significant decrease in fibrinogen levels was observed in non-smokers (–32 mg/dl, 95% CI –63 to –1). No significant changes in WBC, neutrophil, and fibrinogen levels were found in smokers. Sixteen hours after welding exposure, CRP levels were found to be significantly increased in both non-smokers and smokers (0.90 mg/l, 95% CI 0.17 to 1.64). PM_2.5 concentrations were found to be significantly associated with absolute neutrophil counts in non-smokers, and CRP levels in both non-smokers and smokers.

Conclusions: High levels of welding fume exposure induce acute systemic inflammation in a relatively young, healthy working population. These results also suggest that smoking may modify the effect of welding fume exposure on specific inflammatory markers.

     

The prevalence of inflammatory periodontitis is negatively associated with serum antioxidant concentrations

Chronic periodontitis is an inflammatory disease that affects the supporting tissues of the teeth. It is initiated by specific bacteria within the plaque biofilm and progresses due to an abnormal inflammatory-immune response to those bacteria. Periodontitis is the major cause of tooth loss and is also significantly associated with an increased risk of stroke, type-2 diabetes and atheromatous heart disease. Oxidative stress is reported in periodontitis both locally and peripherally (serum), providing potential mechanistic links between periodontitis and systemic inflammatory diseases. It is therefore important to examine serum antioxidant concentrations in periodontal health/disease, both at an individual species and total antioxidant (TAOC) level. To determine whether serum antioxidant concentrations were associated with altered relative risk for periodontitis, we used multiple logistic regression for dual case definitions (both mild and severe disease) of periodontitis in an analysis of 11,480 NHANES III adult participants (>20 y of age). Serum concentrations of vitamin C, bilirubin, and TAOC were inversely associated with periodontitis, the association being stronger in severe disease. Vitamin C and TAOC remained protective in never-smokers. Higher serum antioxidant concentrations were associated with lower odds ratios for severe periodontitis of 0.53 (CI, 0.42,0.68) for vitamin C, 0.65 (0.49,0.93) for bilirubin, and 0.63 (0.47,0.85) for TAOC. In the subpopulation of never-smokers, the protective effect was more pronounced: 0.38 (0.26,0.63, vitamin C) and 0.55 (0.33,0.93, TAOC). Increased serum antioxidant concentrations are associated with a reduced relative risk of periodontitis even in never-smokers.     

Citrulline is an effective arginine precursor in enterally fed neonatal piglets

Although neonatal piglets can synthesize some arginine from proline, there is a limit to this synthesis, and piglets fed an arginine-deficient diet have diminished whole-body arginine status. To help elucidate where the limitation in arginine synthesis may occur, our objective was to determine the most effective arginine precursor in 1-wk-old enterally fed piglets. Piglets were administered either an arginine-deficient (basal) diet [1.15 mmol arginine/(kg.d)] or the basal diet supplemented with equimolar [9.18 mmol/(kg.d)] amounts of proline (+Pro), ornithine (+Orn), citrulline (+Cit) or arginine (+Arg) for 5 d (n = 5/diet). Daily blood samples were taken and indicators of whole-body arginine status including plasma amino acid, ammonia, and urea concentrations were measured. A primed, constant intragastric (i.g.) infusion of l-[U-(14)C]proline was given to measure the proline to arginine conversion, and intravenous (i.v.) and i.g. infusions of l-[guanido-(14)C]arginine were given to determine arginine flux and to quantify the splanchnic extraction of dietary arginine. Piglets fed the +Cit and +Arg diets had lower plasma ammonia and urea concentrations (P < 0.05) and higher plasma arginine concentrations (P < 0.0001) and arginine fluxes (P < 0.05) than piglets fed the other 3 diets. Piglets fed +Cit and +Arg had a lower proline to arginine conversion (P < 0.05). During first-pass splanchnic metabolism, 52% of the dietary arginine was extracted, and this extraction was not affected by whole-body arginine status (P > 0.05). These data indicate that citrulline, but not ornithine or proline, is an effective arginine precursor, and that either citrulline formation or availability appears to limit arginine synthesis in neonatal piglets.