Metabolic profiles in the response to supplementation with composite antimicrobial peptides in piglets challenged with deoxynivalenol.

Deoxynivalenol (DON) causes various toxic effects in human and animals. However, our previous studies have shown that composite antimicrobial peptides (CAP) can have a protective effect in piglets challenged with DON. This study was conducted to evaluate the effect of the CAP GLAM 180# on the metabolism of piglets challenged with DON using a nuclear magnetic resonance (NMR)-based metabolomics approach. A total of 28 individually housed piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned into 4 treatment groups (7 pigs/treatment) based on a 2 × 2 factorial arrangement that were fed, respectively, a basal diet (NC), basal diet + 0.4% CAP (basal + CAP), basal diet + 4 mg/kg DON (basal + DON), and basal diet + 4 mg/kg DON + 0.4% CAP (DON + CAP). A 7-d adaptation period was followed by 30 d of treatment. Blood samples were then collected for metabolite analysis by proton NMR (H-NMR) spectroscopy and liquid chromatography tandem mass spectrometry (LC-MS/MS). The combined results of H-NMR spectroscopy and LC-MS/MS showed that DON increased ( < 0.05) the serum concentrations of low-density lipoprotein, glycoprotein, urea, trimethylamine-N-oxide (TMAO), and lactate as well as those of almost all essential AA and some nonessential AA but decreased the concentrations of high-density lipoprotein (HDL), unsaturated lipids, citrate, choline, and fumarate compared with those in NC treatment ( < 0.05). There was a significant interaction effect ( < 0.05) of supplementation with DON and CAP on some metabolites showed that the serum concentrations of HDL, unsaturated lipids, Pro, citrate, and fumarate were greater ( < 0.05) whereas those of glycoprotein, urea, TMAO, Gly, and lactate were lower in the DON + CAP treatment compared with those in the basal + DON treatment ( < 0.05). These findings indicated that DON causes disturbances in AA, lipid, and energy metabolism and that CAP could partially attenuate the above metabolic disturbances induced by DON.

Effects of composite antimicrobial peptides in weanling piglets challenged with deoxynivalenol: I. Growth performance, immune function, and antioxidation capacity.

The mycotoxin deoxynivalenol (DON) is a food contaminant that leads to reduced feed intake and reduced BW gain, as well as organ impairment. On the other hand, antimicrobial peptides have been shown to have positive effects on growth performance, nutrient digestibility, and immune function. The purpose of this study was to investigate the protective effects of composite antimicrobial peptides (CAP) on piglets challenged with DON. After a 7-d adaptation period, 28 individually housed piglets (Duroc × Landrace × Large Yorkshire) weaned at 28 d of age were randomly assigned to receive 1 of 4 treatments (7 pigs/treatment): negative control, basal diet (NC), basal diet + 0.4% CAP (CAP), basal diet + 4 mg/kg DON (DON), and basal diet + 4 ppm DON + 0.4% CAP (DON + CAP). On d 15 and 30 after the initiation of treatment, blood samples were collected for the determination of blood profile. Piglets were monitored for 30 d to assess performance and then were slaughtered to obtain organs for the determination of the relative weight of organs. The results showed that dietary supplementation with DON decreased (P < 0.05) ADFI, ADG, and G:F, whereas dietary supplementation with CAP improved ADG and G:F (P < 0.05). The relative weight of the kidney and pancreas was greater and the relative weight of the spleen was lighter in the DON treatment than in the other 3 treatments (P < 0.05). There were no effects (P > 0.05) on other relative weights of viscera, except the relative weight of the gallbladder, but the diamine oxidase activity in the liver decreased in DON-treated piglets (P < 0.05). Piglets in the DON treatment had increased serum concentrations of alkaline phosphatase, alanine transaminase, and aspartate aminotransferase and a dramatic decrease in total protein (P < 0.05), whereas there were no differences (P > 0.05) between the DON + CAP treatment and the other treatments. The DON treatment decreased the numbers of red blood cells and platelets, as well as the serum catalase concentrations, and decreased the serum concentrations of H2O2, maleic dialdehyde, and nitric oxide (P < 0.05). The numbers of platelets and thrombocytocrit, as well as the serum concentrations of catalase, were greater, whereas the maleic dialdehyde concentrations were decreased, in both the CAP and DON + CAP treatments compared with the other treatments (P < 0.05). Compared with the control treatment, DON decreased peripheral lymphocyte proliferation on d 15, whereas supplementation with CAP increased it on d 15 and 30 (P < 0.05). These findings indicate that CAP could improve feed efficiency, immune function, and antioxidation capacity and alleviate organ damage, and thus, it has a protective effect in piglets challenged with DON.

Effect of dietary arginine and N-carbamoylglutamate supplementation on reproduction and gene expression of eNOS, VEGFA and PlGF1 in placenta in late pregnancy of sows.

The objectives of this study were to investigate the potential mechanisms of dietary arginine (Arg) and N-carbamoylglutamate (NCG) supplementation on reproductive performance of sows. Twenty-seven crossbred (Landrace×Large White) sows with similar body weight and parity at day (90±1) of gestation were assigned randomly into 3 groups (n=9) control group, Arg group, NCG group, and fed with the following diets: a control diet, and the control diet supplemented with 1.0% Arg or 0.1% NCG. Litter size was recorded. Blood samples were obtained for biochemical analyses. Placenta chorioallantoic membrane tissue collected immediately after birth to preserve in RNA stabilizer for mRNA analysis of endothelial nitric oxide synthase (eNOS), endothelial growth factor a (VEGFA) and placenta growth factor 1 (PlGF1) by real time-PCR. The results showed that compared with the control group, the average birth weight of all piglets born alive were 16.2% and 14.3% higher in the Arg and NCG groups (P<0.05), respectively; plasma VEGFA was higher in the Arg group (P<0.05). The expression of VEGFA in the allantochorion tissue of the NCG-supplemented group was higher (P<0.01), and tended to be higher in the Arg-supplemented group (0.05

Effects of oral supplementation with glutamate or combination of glutamate and N-carbamylglutamate on intestinal mucosa morphology and epithelium cell proliferation in weanling piglets.

To evaluate the effects of glutamate (Glu) or combination of Glu and N-carbamylglutamate (NCG) on intestinal mucosa morphology and epithelium cell proliferation, 18 piglets weaned at 21 d (BW 5.56 ± 0.51 kg) were grouped into 3 treatments and fed one of the following diets for 20 d: a standard diet (SD), SD+Glu(1%), or SD+Glu(1%)+NCG(0.05%). All the piglets were killed for intestinal mucosa collection, and real-time PCR was used to detect mRNA abundance of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and ß-catenin. The results showed that compared with the control group, adding Glu or Glu+NCG to the diet resulted in a higher villus height and mucosal thickness (P < 0.05) in the jejunum. However, the villus height/crypt depth ratio was unaltered. The RT-PCR results showed that Glu+NCG significantly increased PCNA mRNA abundance in both jejunum and ileum (P < 0.05), while they also significantly increased ß-catenin and VEGF mRNA abundance in ileum (P < 0.05). Only Glu increased PCNA mRNA abundance in the jejunum (P < 0.05) and ß-catenin mRNA in the jejunum (P < 0.05). These results indicated that oral supply of Glu improved intestinal mucosa morphology, and combined Glu and NCG may have favorable effects on intestinal epithelium cell proliferation than Glu alone.

Inhibitions of vascular endothelial growth factor expression and foam cell formation by EGb 761, a special extract of Ginkgo biloba, in oxidatively modified low-density lipoprotein-induced human THP-1 monocytes cells.

It has been reported that oxidatively modified low-density lipoprotein (Ox-LDL) involvement with vascular endothelial growth factor (VEGF) and foam cell formation play an important role in atherosclerosis (AS). Protective effects of Ginkgo biloba extract (EGb 761) have been identified for some cardiovascular and neurological disorders. The aim of this study was to investigate whether Ox-LDL regulates VEGF expression in human THP-1 monocytes, as well as the effect of EGb 761 on VEGF expression and the formation of foam cells. After exposure to Ox-LDL alone or in combination with EGb 761 for up to 48h, cell viability was measured using the MTT assay. VEGF protein content in the supernatant was analyzed by enzyme-linked immunosorbent assay (ELISA). VEGF mRNA was determined by real-time PCR. To determine the effect of EGb 761 on foam cell formation, an Ox-LDL-induced foam cell model was used. Ox-LDL inhibited the growth of THP-1 cells and EGb 761 increased the cell survival rate. Ox-LDL markedly increased VEGF expression in THP-1 cells in a time- and concentration-dependent manner, which was significantly suppressed by EGb 761. EGb 761 also inhibited monocyte/macrophage-derived foam cell formation. These results suggest that Ox-LDL is involved in the development of human AS through VEGF induction in monocytes, and that EGb 761 prevents in vitro atherogenesis, probably via downregulation of VEGF expression in monocytes and inhibition of monocyte/macrophage-derived foam cell formation. The findings suggest a mechanism for the in vivo anti-AS effect of EGb 761 and support its potential clinical use in AS.

Dietary supplementation with Chinese herbal powder enhances ileal digestibilities and serum concentrations of amino acids in young pigs.

This study was designed to determine the effect of ultra-fine Chinese herbal powder as a dietary additive on serum concentrations and apparent ileal digestibilities (AID) of amino acids (AA) in young pigs. In Experiment 1, 60 Duroc x Landrace x Yorkshire piglets weaned at 21 days of age were randomly assigned to one of three treatments, representing supplementation with 0 or 2 g/kg of the powder, or 0.2 g/kg of colistin (an antibiotic) to corn- and soybean meal-based diets (n = 20 per group). Blood samples from five piglets per group were collected on days 7, 14, and 28 to determine serum AA concentrations. In Experiment 2, 12 barrows with an average initial body weight of 7.64 kg were randomly assigned to one of the three dietary treatments, followed by surgical placement of a simple T-cannula at the terminal ileum. All of the diets contained 0.1% titanium oxide as a digestibility marker. The samples of terminal ileal digesta were collected on day 7 for determining AID of AA. Results show that dietary supplementation with the herbal powder increased (P < 0.05) serum concentrations and AID of most AA by 10-50% and 10-16%, respectively. As an indicator of improved intestinal function, AID values of calcium were also enhanced in piglets supplemented with the herbal powder. Dietary supplementation of colistin increased serum concentrations and AID values of some AA by 8-44% and 10-15%, respectively, in comparison with the non-supplemented group. These novel findings demonstrate that the herbal powder can enhance the digestibility of dietary protein and the intestinal absorption of AA into the systemic circulation in post-weaning pigs, therefore providing a new mechanism for its growth- and immunity-promoting efficacy.

Dietary supplementation with Astragalus polysaccharide enhances ileal digestibilities and serum concentrations of amino acids in early weaned piglets.

Two experiments were conducted to evaluate the effects of dietary supplementation with Astragalus polysaccharide (APS) on growth performance, apparent ileal digestibilities (AID) of amino acids (AA), and their serum concentrations in early weaned piglets. In Exp. 1, 60 pigs were weaned at 21 days of age (BW 7.35 +/- 0.23 kg) and allocated to three treatments (20 pigs/treatment), representing supplementing 0.0% (control), 0.02% colistin (antibiotic), or 0.1% APS to a corn- and soybean meal-based diet. Average daily gain (ADG), average daily feed intake (ADFI), and feed/gain ratio (F/G) were measured weekly. Blood samples were obtained from five pigs selected randomly from each treatment for the measurement of serum free AA concentrations on days 7, 14, and 28. In Exp. 2, 12 pigs were weaned at 21 day of age (BW 7.64 +/- 0.71 kg), assigned to three treatment groups as in Exp. 1, and surgically fitted with a simple T-cannula at the terminal ileum. Ileal digesta samples were obtained for the measurement of AID of AA on days 7, 14 and 28. Dietary APS did not affect ADFI, but enhanced (P < 0.05) ADG by 11 and 4.4%, and improved F/G by 5.6 and 8.4%, respectively, compared with the control and antibiotic groups. Addition of APS to the diet increased AID and serum concentrations of most nutritionally essential and non-essential AA (including arginine, proline, glutamate, lysine, methionine, tryptophan, and threonine) on days 14 and 28. Circulating levels of total AA were affected by the age of pigs and treatment x time interaction. Collectively, these findings indicate that APS may ameliorate the digestive and absorptive function and regulate AA metabolism to beneficially increase the entry of dietary AA into the systemic circulation, which provide a mechanism to explain the growth-promoting effect of APS in early weaned piglets.

Effect of dietary oligochitosan supplementation on ileal digestibility of nutrients and performance in broilers.

The effect of dietary chitosan oligosaccharides (COS) supplementation on ileal digestibilities of nutrients and performance in broilers was assessed by feeding graded levels (0, 50, 100, 150 mg/kg) of COS. Two thousand four hundred male commercial Avian broilers (1-d-old) were assigned randomly to 5 dietary treatment groups (60 birds per pen with 8 pens per treatment). Diet A was a typical corn- and soybean meal-based diet supplemented with 6 mg/kg of an antibiotic flavomycin (positive control). Diet B was the basal diet without any supplement. Diets C, D, and E were formulated by adding 50, 100, and 150 mg/kg of COS to the basal diet, respectively. On the morning of d 21 and 42, 64 birds (8 per pen with 8 pens per treatment) from the growth trial for each age group were killed by cervical dislocation for determination of the ileal digestibilities of nutrients. Dietary supplementation with COS and antibiotic enhanced (P < 0.05) the ileal digestibilities of DM, Ca, P, CP, and all amino acids (except for alanine in the 21-d-old birds or phenylalanine, glutamate, and glycine for the 42-d-old birds). Feed efficiency was improved (P < 0.05) in response to dietary supplementation of an antibiotic or COS (150 mg/kg for d 1 to 21, and 100 and 150 mg/kg for d 21 to 42). The results demonstrate for the first time to our knowledge that dietary COS supplementation was effective in increasing the ileal digestibilities of nutrients and feed efficiency in broilers. Our findings may explain a beneficial effect of COS on chicken growth performance.

VEGF induced hyperpermeability in bovine aortic endothelial cell and inhibitory effect of salvianolic acid B.

AIM: To examine the effect of recombinant human vascular endothelial growth factor (VEGF) on the low density lipoprotein (LDL) permeability of bovine aortic endothelial cells (BAEC) and the inhibitory effect of salvianolic acid B in vitro. METHODS: The confluence BAEC monolayers were cultured with normal medium and with medium containing VEGF or salvianolic acid B at various concentrations and for various time periods. The iodine labeled LDL flux across the monolayers was then performed, and radioactivity was measured by SN-695 automatic liquid scintillation counter. RESULTS: Addition of purified human recombinant VEGF to the BAEC monolayers could significantly increase the permeability of the monolayer to 125I-LDL (P < 0.01). The permeability-increasing activity of VEGF on the BAEC monolayers was both dose and time dependent. Salvianolic acid B could markedly inhibit the VEGF-induced hyperpermeability in BAECs (P < 0.01). CONCLUSION: VEGF plays a role in the formation and development of atherosclerosis, and salvianolic acid B has inhibitory effect on VEGF-induced hyperpermeability in BAEC.

Three new saponins from Tribulus terrestris.

Three new steroidal saponins 1-3, together with five known steroidal saponins, L-mannitol and an inorganic salt were isolated from Tribulus terrestris L. (Zygophyllaceae). The structures of the new steroidal saponins were elucidated as hecogenin 3-O-beta-xylopyranosyl(1-->3)-beta-glucopyranosyl(1-->4)-beta-galactopyr anoside (1), hecogenin 3-O-beta-glucopyranosyl(1-->2)-beta-glucopyranosyl(1-->4)- beta-galactopyranoside (2) and 3-O-[beta-xylopyranosyl(1-->2)-[beta-xylopyranosyl(1-->3)]-beta- glucopyranosyl(1-->4)-[alpha-rhamnopyranosyl(1-->2)]-beta-galactopyranos yl]- 26-O-beta-glucopyranosyl-22-methoxy-(3 beta,5 alpha,25R)-furostan-3,26-diol (3). Structure elucidation was accomplished by 1D and 2D NMR spectra (13C-1H COSY, HMQC, HMBC, 1H-1H COSY, TOCSY, and NOESY), mass spectrometry (FABMS, ESIMS) and chemical methods.

Antagonistic effects of Ginkgo biloba extract on adhesion of monocytes and neutrophils to cultured cerebral microvascular endothelial cells.

AIM: To study the action of Ginkgo biloba extract (GbE) on tumor necrosis factor (TNF-alpha)-induced adhesion of monocytes (Mon) and neutrophils (Neu) to cultured cerebral microvascular endothelial cells. METHODS: TNF-alpha-induced endothelial adhesivity toward Mon and Neu was studied using bovine cerebral microvascular endothelial cells (BCMEC) in vitro. The number of Mon and Neu adhering to the BCMEC monolayers was determined by flow cytometry. RESULTS: Pretreatment of BCMEC with TNF-alpha increased Mon and Neu adhesion to BCMEC from 12.5% +/- 0.2% to 31.3% +/- 0.5% and from 13.8% +/- 0.4% to 32.1% +/- 0.5%, respectively. GbE (1-100 mg.L-1) inhibited the effect of TNF-alpha in a concentration-dependent manner. E-selectin mAb (1 mg.L-1) blocked Mon and Neu adhesion to BCMEC induced by TNF-alpha. CONCLUSION: The inhibition of GbE on Mon and Neu adhesion to BCMEC was mediated through the suppression of E-selection expression.

[An preliminary study on hepato-protective action of seed oil of Hippophae rhamnoides L. (HR) and mechanism of the action].

The seed oil of Hippophae rhamnoides markedly inhibits the rise of MDA in liver of mice and rats induced by CCl4, AAP and ethyl alcohol, decreases significantly the activity of SGPT and SGOT, and markedly checks the depletion of GSH in liver of mice induced by AAP. The microscopic and electron-microscopic examination has shown that the seed oil can lighten liver injury.