Acid-supplemented Diet Research Articles

Effects of low protein diets with amino acids supplementation on biochemical and faeces parameters in weaned piglets

Effects of low protein diets with amino acids supplementation on biochemical and faeces parameters in weaned piglets

Determination of sialic acid in dietary supplement by liquid chromatography tandem-mass spectrometry (LC-MS/MS)

Simultaneous determination of sialic acids (N-acetyl neuraminic acid - NANA and N-glycolyl neuraminic acid - NGNA) in dietary supplement matrices by liquid chromatography tandem-mass spectrometry (LC-MS/MS) had been developed. Sialic acids were separated by using a C18 column (75 mm x 3.9 mm, 2.5 µm) and isocratic mobile phase of acid formic 0.1 %: acetonitrile (80:20, v/v). Sialic acids were totally hydrolyzed by formic acid 1M at 80oC in 2 hours before being analyzed by LC-MS/MS. It was proved that the method with the linearity range of 0.04 - 4.25 µg/ml had highly specificity, selectivity, repeatability, and recovery rate, that well satisfied AOAC requirements. The method has been applied to detect sialic acids in commercial dietary supplements.

Ingestion of difructose anhydride III partially suppresses the deconjugation and 7α-dehydroxylation of bile acids in rats fed with a cholic acid-supplemented diet.

Difructose anhydride III (DFAIII) is a prebiotic involved in the reduction of secondary bile acids (BAs). We investigated whether DFAIII modulates BA metabolism, including enterohepatic circulation, in the rats fed with a diet supplemented with cholic acid (CA), one of the 12α-hydroxylated BAs. After acclimation, the rats were fed with a control diet or a diet supplemented with DFAIII. After 2weeks, each group was further divided into two groups and was fed diet with or without CA supplementation at 0.5g/kg diet. BA levels were analyzed in aortic and portal plasma, liver, intestinal content, and feces. As a result, DFAIII ingestion reduced the fecal deoxycholic acid level via the partial suppression of deconjugation and 7α-dehydroxylation of BAs following CA supplementation. These results suggest that DFAIII suppresses production of deoxycholic acid in conditions of high concentrations of 12α-hydroxylated BAs in enterohepatic circulation, such as obesity or excess energy intake. Abbreviation: BA: bile acid; BSH: bile salt hydrolase; CA: cholic acid; DCA: deoxycholic acid; DFAIII: difructose anhydride III; MCA: muricholic acid; MS: mass spectrometry; NCDs: non-communicable diseases; LC: liquid chromatography; SCFA: short-chain fatty acid; TCA: taurocholic acid; TCDCA: taurochenodeoxycholic acid; TDCA: taurodeoxycholic acid; TUDCA: tauroursodeoxychlic acid; TαMCA: tauro-α-muricholic acid; TβMCA: tauro-β-muricholic acid; TωMCA: tauro-ω-muricholic acid.

Amino Acids and Hypertension in Adults.

Accumulating evidence suggests a potential role of dietary protein among nutritional factors interfering with the regulation of blood pressure. Dietary protein source (plant versus animal protein), and especially, protein composition in terms of amino acids has been postulated to interfere with mechanisms underlying the development of hypertension. Recently, mounting interest has been directed at amino acids in hypertension focusing on habitual dietary intake and their circulating levels regardless of single amino acid dietary supplementation. The aim of the present review was to summarize epidemiological evidence concerning the connection between amino acids and hypertension. Due to the large variability in methodologies used for assessing amino acid levels and heterogeneity in the results obtained, it was not possible to draw robust conclusions. Indeed, some classes of amino acids or individual amino acids showed non-causative association with blood pressure as well as the incidence of hypertension, but the evidence was far from being conclusive. Further research should be prompted for a thorough understanding of amino acid effects and synergistic actions of different amino acid classes on blood pressure regulation.

High-yielding lactating sows maintained a high litter growth when fed reduced crude protein, crystalline amino acid-supplemented diets

The study was conducted to determine the optimal standardized ileal digestible (SID) CP level to maximize litter gain in high-yielding lactating sows when feeding reduced CP, crystalline amino acid-supplemented diets. In total, 520 Landrace × Yorkshire sows (parity 1 to 5) were allocated to 1 of 6 diets from day 2 post-partum when litters were standardized to 14 piglets. All diets were formulated to meet or slightly exceed Danish recommendations for Lys, Met, Met + Cys, Thr and Trp using crystalline amino acids. Diets contained 116, 126, 132, 138, 144, and 153 g SID CP per kg (as-fed). Sow body weight, back fat thickness, and litter weight were recorded at litter standardization and at weaning. On a subsample of 72 sows (parity 2 to 4), milk samples were collected at day 3, 10 and 17 post-partum, and blood was sampled at day 10 and 17 post-partum. All data were subjected to ANOVA using the MIXED procedure in SAS. Litter size at weaning was not affected by the dietary CP content (13.0 ± 0.06; P = 0.40), nor was litter ADG (3.13 ± 0.03 kg/d; P = 0.52), nor milk contents of fat and lactose (P > 0.05). Likewise, the daily changes in sow body weight (−0.58 ± 0.04 kg/d, P = 0.44) and back fat thickness (−0.15 ± 0.004 mm/d, P = 0.20) were similar across dietary treatments. Milk protein (P < 0.01), milk Lys (P < 0.05), and other essential amino acids in milk protein were reduced at a SID CP concentration of 116 g/kg. Plasma concentrations of NEFA and creatinine were not affected by dietary treatment (P > 0.05), but concentrations of plasma urea nitrogen were elevated (P < 0.001) as dietary CP content increased. In conclusion, sows were able to maintain a high litter growth when fed reduced CP, crystalline amino acid-supplemented diets. Though, it is not recommended to feed the lactating sows less than 126 g SID CP/kg (as-fed).

Moderate maternal folic acid supplementation ameliorates adverse embryonic and epigenetic outcomes associated with assisted reproduction in a mouse model.

Could clinically-relevant moderate and/or high dose maternal folic acid supplementation prevent aberrant developmental and epigenetic outcomes associated with assisted reproductive technologies (ART)? Our results demonstrate dose-dependent and sex-specific effects of folic acid supplementation in ART and provide evidence that moderate dose supplements may be optimal for both sexes. Children conceived using ART are at an increased risk for growth and genomic imprinting disorders, often associated with DNA methylation defects. Folic acid supplementation is recommended during pregnancy to prevent adverse offspring outcomes; however, the effects of folic acid supplementation in ART remain unclear. Outbred female mice were fed three folic acid-supplemented diets, control (rodent daily recommended intake or DRI; CD), moderate (4-fold DRI; 4FASD) or high (10-fold DRI; 10FASD) dose, for six weeks prior to ART and throughout gestation. Mouse ART involved a combination of superovulation, in vitro fertilisation, embryo culture and embryo transfer. Midgestation embryos and placentas (n = 74-99/group) were collected; embryos were assessed for developmental delay and gross morphological abnormalities and embryos and placentas were examined for epigenetic defects. We assessed methylation at four imprinted genes (Snrpn, Kcnq1ot1, Peg1 and H19) in matched midgestation embryos and placentas (n = 31-32/group) using bisulfite pyrosequencing. In addition, we examined genome-wide DNA methylation patterns in placentas (n = 6 normal placentas per sex/group) and embryos (n = 6 normal female embryos/group; n = 3 delayed female embryos/group) using reduced representation bisulfite sequencing (RRBS). Moderate, but not high dose supplementation, was associated with a decrease in the proportion of developmentally delayed embryos. Although moderate dose folic acid supplementation reduced DNA methylation variance at certain imprinted genes in embryonic and placental tissues, high dose supplementation exacerbated the negative effects of ART at imprinted loci. Furthermore, folic acid supplements resolved female-biased aberrant imprinted gene methylation. Supplementation was more effective at correcting ART-induced genome-wide methylation defects in male versus female placentas; however, folic acid supplementation also led to additional methylation perturbations which were more pronounced in males. The RRBS data from this study have been submitted to the NCBI Gene Expression Omnibus under the accession number GSE123143. Although the combination of mouse ART utilised in this study consisted of techniques commonly used in human fertility clinics, there may be species differences. Therefore, human studies, designed to determine the optimal levels of folic acid supplementation for ART pregnancies, and taking into account foetal sex, are warranted. Taken together, our findings support moderation in the dose of folic acid supplements taken during ART. This work was funded by the Canadian Institutes of Health Research (FDN-148425). The authors declare no conflict of interest.

Boswellic Acid Composition of Frankincense Dietary Supplements and Correlation to Cytotoxic Efficacy against Treatment‐Resistant Triple Negative Breast Cancer Cells

Frankincense is an oleogum resin from Boswellia trees of the Burseraceae family. The oleogum resin contains 15–20 % triterpenic acids, including therapeutically active boswellic acids [1]. Extracts of oleogum resins obtained from Boswellia serrata have been traditionally used in Indian folk medicine (Ayurveda) for the treatment of various chronic inflammatory diseases such as rheumatoid arthritis. Furthermore, boswellic acids are known to possess anticancer efficacy against various human cancer types, e.g. gliomas, prostate cancer, or leukemia [2–4]. The aim of the present study was to analyze the contents of boswellic acids in dietary supplements from various Boswellia species grown in different areas and to correlate the concentrations of distinct boswellic acids to the extracts' cytotoxic efficacy against chemoresistant triple‐negative human breast cancer cells. Accordingly, a new method to quantify a large variety of pentacyclic triterpenic acids was developed. Dietary supplements from various Boswellia species Boswellia serrata, Boswellia sacra, Boswellia carterii, and others were extracted with methanol and analyzed by high performance reversed‐phase liquid chromatography coupled with tandem mass spectrometric detection (HPLC‐MS/MS). Besides, the effects of the extracts against the treatment‐resistant, triple negative human breast cancer cell line MDA‐MB‐231 were investigated in vitro by using a cell viability assay. The extracts exhibited potent cytotoxic activity against the human breast cancer cell line. Correlations between the boswellic acid contents and the cytotoxic efficacy were determined by multiple linear regression and principal component regression. This approach allowed not only the correlation of complex phytochemical mixtures to desired biomedical effects, but it likewise provided evidence for key compounds governing the therapeutic efficacy. Hence, the study provides new insights into differential chemical composition patterns of frankincense dietary supplements from Boswellia species grown in different regions. Besides, the study identified distinct components correlating positively with the desired cytotoxic activity. Thus, the identified key compounds deserve further studies either as chemically pure substances or as defined chemical mixtures permitting development of the most active ingredients as potential anticancer drug candidates.Support or Funding InformationThis work was supported by the Academic Center for Complementary and Integrative Medicine (AZKIM), State Ministry of Baden‐Württemberg for Sciences, Research and Arts.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Quantification of omega-3 fatty acids in dietary supplements and cooking products available on the polish market by thin-layer chromatography–densitometry

The effect of omega-3 fatty acids in the human body is multidirectional. They are used in cancer and cardiovascular disease prevention, to stabilize blood pressure, to improve immune function, and in the treatment of concentration disorders. Omega-3 acids, especially eicosapentaenoic and docosahexaenoic acids, are needed for proper human growth. There are several groups of pharmaceutical products containing these compounds, available in pharmacies and markets. Due to a high accessibility of various consumable products such as dietary supplements or food, their quality monitoring is advisable. Therefore, analytical procedures are needed, which will allow the quality of these products to be fast and accurately defined. In our work, thin-layer chromatography (TLC) with densitometric detection was designed for the identification and quantification of the omega-3 fatty acids. The stationary phase constituted TLC silica gel plates activated by AgNO3 solution. As the mobile phase, a mixture of toluene—n-propanol—acetic acid (20:2:0.1, v/v) was used. Detection was performed at 520 nm, after staining with iodine vapor. The method was validated according to the International Conference on Harmonisation (ICH) guidelines. The correctness of the results guarantees good precision and satisfactory recovery (96.59–103.20%). The presented analytical method was used for the determination of omega-3 fatty acids in selected dietary supplements and cooking oils giving satisfactory results.

Enantiomeric determination of α-lipoic acid in urine by LC/MS/MS

An analytical method for the enantiomeric determination of α-lipoic acid in human urine was developed to evaluate the pharmacokinetics of α-lipoic acid, an ingredient in health food. Urine samples were collected over time after administering α-lipoic acid dietary supplement to healthy subjects. The samples were cleaned up by solid-phase extraction using an Oasis® MAX cartridge. In the LC/MS/MS method, CHIRALPAK AD-3R was used as the chiral separation column and acetonitrile-methanol-formic acid (10 mM) (25:25:50, v/v/v) was used as the mobile phase. 13C4 1,2,5,6-d-l-α-Lipoic acid was used as the internal standard. MS/MS was performed by electrospray ionization (ESI) in the negative ion mode using two monitoring ion transitions (m/z 205.0 → 170.9 and m/z 209.0 → 174.9). A calibration curve was prepared in the concentration range of 0.5–100 ng/mL for each enantiomer (r > 0.9999). The limit of detection (LOD, S/N = 3) and the limit of quantification (LOQ, S/N > 10) were 0.1 ng/mL and 0.5 ng/mL, respectively. The intra-day and inter-day accuracy of α-lipoic acid enantiomers at the LOQ level (0.5 ng/mL), the low concentration level (5 ng/mL), the middle concentration level (50 ng/mL), and the high concentration level (100 ng/mL) ranged from 93.7 to 103.1%. The intra-day and inter-day precision were ≦ 6.94% and ≦ 7.05%, respectively. Calculating the mean values of pharmacokinetic parameters revealed that the AUC and Cmax values of d-α-lipoic acid were statistically significantly higher than those of l-α-lipoic acid (p < 0.05). It was suggested that l-α-lipoic acid is more bioavailable than d-α-lipoic acid despite individual differences in excretion rate.

PM2.5 exposure induces systemic inflammation and oxidative stress in an intracranial atherosclerosis rat model.

Exposure to airborne particle (PM2.5 ) is a risk factor for intracranial atherosclerosis (ICA). Because of the established role of systemic inflammation and oxidative stress by PM2.5 , we determined whether these processes account for PM2.5 -mediated ICA, and also whether omega-3 fatty acid (O3FA) dietary supplementation could attenuate them. Adult Sprague-Dawley rats were exposed to filtered air (FA) or PM2.5 and fed either a normal chow diet (NCD) or a high-cholesterol diet (HCD), administered with or without O3FA (5 mg/kg/day by gavage) for 12 weeks. The lumen and thickness of the middle cerebral artery (MCA) were assessed. Serum tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin-1β (IL-1β), and interferon gamma (IFN-γ) were detected by ELISA. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) activity, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) activity, mRNA levels of Nrf2, HO-1, NQO-1, and protein level of NOX subunit gp91 were quantified to determine the oxidative profile of brain vessels. PM2.5 increased (P < .05) ICA, especially in the HCD group; elevated serum TNF-α, IL-6, IL-1β, and IFN-γ; increased cerebrovascular ROS, MDA, NOX activity, and gp91 protein levels; and decreased cerebrovascular SOD activity. Nrf2, HO-1, and NQO-1 mRNA levels were upregulated (P < .05) by PM2.5 exposure, especially in the HCD group. O3FA attenuated (P < .05) PM2.5 -induced systemic inflammation, vascular oxidative injury, and ICA. PM2.5 exposure induced systemic inflammation, cerebrovascular oxidative injury, and ICA in rats with HCD. O3FA prevented ICA development, and may therefore exert a protective effect against the atherogenic potential of PM2.5 .

Multiple feedings enhance the growth performance and feed efficiency of juvenile Litopenaeus vannamei when fed a low-fish meal amino acid-supplemented diet

In the coming years, supplementation of crystalline amino acids (CAAs) will become a standard practice driven by the trend towards low-fish meal diets. We have evaluated the effect of feeding juvenile L. vannamei several times a day versus two (2×) and four (4×) times using a low-fish meal diet supplemented with CAAs. A total of 1632 shrimp of 1.06 ± 0.16 g were stocked in 16 outdoor tanks of 1 m3 under 100 shrimp/m2. Shrimp were either fed manually, 2× or 4× daily, using one feeding tray per tank, or fed with an automatic feeder to deliver multiple (10) meals during the day or during the day and night (D&N) at programmed times. A diet containing 3% fish meal was prepared to contain supplemental levels of CAAs. After 11 weeks of rearing, survival, growth performance, and feed efficiency of shrimp were significantly affected by feeding frequency and time of feeding. Feeding 2× and 4× daily resulted in a lower survival compared to multiple times during the day or D&N. Shrimp body weight was enhanced with an increase in feeding frequency, from 8.74, 10.95, 11.33, and 11.33 g under 2×, 4×, and multiple feedings during the day or during D&N, respectively. FCR was also significantly affected, reducing from a high of 2.46 under 2× to 1.59 under multiple feedings D&N. Our findings indicate that it is more advantageous to feed juvenile L. vannamei multiple times a day when a low-fish meal amino acid-supplemented diet is used.

Dietary Omega-3 Fatty Acid Supplementation Improves Sickle Cell Bone Disease By Affecting Osteoblastogenesis and Adipogenesis

Sickle bone disease (SBD) is a severe and invalidating complication related to recurrent bone vaso-occlusive events affecting patients with sickle cell disease (SCD). In a humanized mouse model for SCD, we previously showed that SBD is sustained by impaired osteoblast function and increased osteoclast activity associated with local up-regulation of pro-inflammatory cytokines and anti-oxidant systems (Dalle Carbonare L., Blood 2015;126:2320-2328). Growing evidence supports a role of ω-3 fatty acid supplementation in improving bone homeostasis (Fallon EM, et al J Surg Res 2014;191:148-155.). A diet enriched with ω-3 fatty acids beneficially impacts SCD inflammatory vasculopathy, and blunts the acute and chronic SCD-related organ damage in humanized SCD mice (Kalish BT et al Haematologica 2015;100:870-880). Here, we sought to compare the dietary effects of ω-6 (soybean oil-based, SD)- vs ω-3 (fish oil-based, FD)-enriched diets on SBD in SCD mice (Hbatm1(HBA)Tow Hbbtm2(HBG1,HBB*)Tow). Treated SCD and control healthy mice (AA, Hbatm1(HBA)Tow Hbbtm3(HBG1,HBB)Tow) (n=6 animals in each group) were exposed to recurrent hypoxia/reoxygenation (rH/R) stress, which closely mimics SBD natural history (Dalle Carbonare L., Blood 2015). In SCD mice, FD prevented rH/R-induced bone loss compared to animals exposed to SD by decreasing osteoclast and increasing osteoblast activities. Up-regulation of molecular osteogenic markers such as Runx2 and Sparc and down-regulation of Rank and RankL, molecular markers of osteoclast recruitment and activity were observed in FD SCD mice exposed to rH/R compared to SD SCD animals. Similar changes, but to a lesser extent, were also observed in AA control mice exposed to rH/R stress. Expression of IL-6 (Il6) and matrix-metalloproteinase-9 (Mmp9) regulators which interact with RankL on osteoclastic activity and bone resorption were studied. In SCD mice, FD markedly reduced the up-regulation of both genes compared to SD SCD animals in conjunction with down-regulation of peroxiredoxin-2 (Prx2) gene expression, an important cytoprotective and antioxidant system. We also evaluated bone adipogenesis, which is believed to be an important contributor to bone impairment in SBD. Bone immunohistochemistry for Peripilin-1 which coats storage lipid droplets revealed increased adipogenesis in SD SCD mice compared to either FD SCD animals or AA controls, in association with downregulation of miR205, which decreases adipogenesis and enhances osteogenic activity. Our data thus indicate that in SCD exposed to rH/R, FD (i) improves osteoblastogenesis; (ii) decreases osteoclast activity; (iii) modulates the bone inflammatory response; iv) reduces adipogenesis. These findings provide new insights on the mechanism of action of ω-3 fatty acid supplementation on the pathogenesis of SBD and strengthen the rationale for ω-3 fatty acid dietary supplementation in SCD as a complementary therapeutic intervention targeting an amplified inflammatory response and sickle cell-related bone impairment. DisclosuresNo relevant conflicts of interest to declare.

Effect of n-3 long-chain polyunsaturated fatty acids supplementation in healthy mothers on DHA and EPA profiles in maternal and umbilical blood: a randomized controlled trial.

Background The objective of the study was to compare the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) dietary supplementation on their concentration in total lipids (TL) and lipid fractions of maternal and umbilical vein (UV) blood. The specific objective was to analyze the impact of EPA and DHA supplementation on pregnancy outcome and neonatal birth weight. Methods Women were randomly single-blinded (randomized controlled trial; ISRCTN36705743) allocated to the group receiving EPA and DHA supplementation (supplemented group) or the group receiving placebo-corn oil (control group) in the time period from January 1st, 2016 until March 1st, 2017. Women in the supplemented group (n=45) took 360 mg EPA and 240 mg DHA daily while controls (n=42) were given a placebo. Maternal and UV bloods were obtained at delivery. After lipid extraction, phospholipids (PL), cholesterol esters (CE), triacylglycerols (TG) and non-esterified fatty acids were separated by thin layer chromatography and analyzed by gas chromatography. Results Higher DHA concentrations in TL (37.24±21.87 mg/L), PL (13.14±8.07 mg/L) and triacylglycerols (2.24±2.21 mg/L) were recorded in mothers from the supplemented group when compared to the study group (TL 21.89±14.53 mg/L; P<0.001; PL 9.33±5.70 mg/L; P=0.013; TG 0.56±0.43 mg/L; P<0.001). Higher DHA concentrations in UV samples were found in TL (11.51±7.34 mg/L), PL (5.29±3.31 mg/L) and triacylglycerols (0.62±0.46 mg/L) from the supplemented groups compared with controls (TL 7.37±3.60 mg/L; P=0.002; PL 3.52±2.19 mg/L; P=0.005; TG 0.40±0.46 mg/L; P=0.035). The ratio of AA:DHA was lower in maternal (2.43) and UV serum (4.0) of the supplemented group than in the control group (maternal 3.85 P<0.001; UV 4.91 P<0.001). Conclusion The study demonstrated the higher ratio of AA/DHA in the control group indicating that pregnant women on the traditional Herzegovina diet need supplementation with DHA and EPA.

Behavioral and cognitive effects of docosahexaenoic acid in drug-naïve children with attention-deficit/hyperactivity disorder: a randomized, placebo-controlled clinical trial.

This study aimed to investigate the efficacy of docosahexaenoic acid (DHA) dietary supplementation on behavior and cognition in school-aged, drug-naïve children with attention-deficit/hyperactivity disorder (ADHD). A total of 50 participants with ADHD aged 7 to 14 were enrolled in a 6-month randomized, placebo-controlled clinical trial and received either DHA or placebo. The primary outcome measure was the change in the ADHD rating scale IV Parent Version-Investigator (ADHD-RS-IV) after 4 and 6months. Secondary outcome measures included Conners Parent Rating Scale-revised, other behavioral rating scales including quality of life and global functioning, and computerized cognitive tasks. Baseline assessment also addressed the blood fatty acids profile. No superiority of DHA supplement to placebo was observed on ADHD-RS-IV, the a priori primary outcome. DHA supplementation showed a significant, nonetheless quite small, effect on children's psychosocial functioning, emotional problems, and focused attention. Neither major nor minor adverse events were reported throughout the trial. This study shows that 6-month DHA supplementation has no beneficial effect on the symptoms of ADHD in school-aged, drug-naïve children with an established diagnosis of ADHD. Nevertheless, the 6months treatment with supplemental DHA appears to have small positive effects on other behavioral and cognitive difficulties, which, in light of the absence of side-effects, could be reasonably followed up in future intervention studies. ( https://clinicaltrials.gov/ct2/show/NCT01796262 : The Effects of DHA on Attention Deficit and Hyperactivity Disorder (DADA)).

Supplementing Oregano Essential Oil in a Reduced-Protein Diet Improves Growth Performance and Nutrient Digestibility by Modulating Intestinal Bacteria, Intestinal Morphology, and Antioxidative Capacity of Growing-Finishing Pigs.

Simple SummarySerious nitrogen pollution and shortage of protein feed resources have constrained the rapid development of the pig industry. Thus, the strategy of using a reduced-protein diet supplemented with amino acids in pig production has been widely accepted. Additionally, antibiotic growth promoters have been widely used in pig production for many years. However, their enormous and uncontrolled use has promoted bacterial resistance, leading to less effective treatment for animal diseases. Accordingly, this study investigated the effects of inclusion of oregano essential oil in a reduced-protein, amino acid-supplemented diet on growth performance, nutrient digestibility, gut health, and antioxidative capacity of growing-finishing pigs as an alternative to antibiotics. Our results suggested that adding oregano essential oil to a reduced-protein diet improved the growth performance and carcass lean percentage of pigs. In addition, long-term supplementation of oregano essential oil to a reduced-protein diet improved the intestinal bacteria, intestinal morphology, and antioxidative capacity of pigs. This study provides theoretical guidance for application of low-protein diet to guide the production of antibiotic-free feed for growing–finishing pigs.This study investigated the effects of supplementing oregano essential oil (OEO) to a reduced-protein diet on growth performance, nutrient digestibility, intestinal bacteria, intestinal morphology, and antioxidative capacity of growing-finishing pigs. Forty-eight barrows were randomly allotted to four treatments including normal-protein diet (NPD), reduced-protein, amino acid-supplemented diet (RPD), the same RPD supplemented with chlortetracycline (RPA), and RPD supplemented with OEO (RPO). The data showed that dietary OEO supplementation increased the average daily gain of pigs compared with NPD and RPD. The gain:feed in RPO- and NPD-fed pigs was higher than those in RPD- and RPA-fed pigs. Increased average daily feed intake and 10th-rib backfat thickness were detected in RPA-fed pigs. Pigs fed the RPO had higher apparent total tract digestibility (ATTD) of crude protein than those fed the other diets. The RPD and RPA treatments showed reduced counts of Lactobacillus spp. in ileal digesta of pigs. The RPA and RPO treatments also showed lower Escherichia coli counts in ileal digesta than the NPD and RPD treatments. Dietary OEO supplementation increased villous height of the jejunum and the ileal and plasma total antioxidative capacity of pigs. In conclusion, dietary OEO supplementation could improve the growth performance and nutrient digestibility of pigs by modulating intestinal bacteria, intestinal morphology, and antioxidative capacity.

Energy utilisation of broiler chickens in response to guanidinoacetic acid supplementation in diets with various energy contents

This experiment was conducted to investigate the effects of guanidinoacetic acid (GAA) on productive performance, intestinal morphometric features, blood parameters and energy utilisation in broiler chickens. A total of 390 male broiler chicks (Ross 308) were assigned to six dietary treatments based on a factorial arrangement (2×3) across 1-15 and 15-35-d periods. Experimental treatments consisted of two basal diets with standard (STD; starter: 12·56 MJ/kg and grower: 12·97 MJ/kg) and reduction (LME; starter: 11·93 MJ/kg and grower: 12·33 MJ/kg) of apparent metabolisable energy (AME) requirement of broiler chickens each supplemented with 0, 0·6 and 1·2 g/kg GAA. Supplemental 1·2 g/kg GAA decreased the negative effects of feed energy reduction on weight gain across starter, growing and the entire production phases (P<0·05). Energy retention as fat and total energy retention were increased when birds received LME diets supplemented with 1·2 g/kg GAA (P<0·05). Net energy for production (NEp) and total heat production increased in birds fed LME diets containing 1·2 g/kg GAA (P<0·05). A significant correlation was observed between dietary NEp and weight gain of broilers (r 0·493; P=0·0055), whereas this relationship was not seen with AME. Jejunal villus height and crypt depth were lower in birds fed LME diets (P<0·05). Serum concentration of creatinine increased in broilers fed LME diets either supplemented with 1·2 g/kg GAA or without GAA supplementation (P<0·05). Supplemental GAA improved performance of chickens fed LME diet possibly through enhanced dietary NEp. The NEp could be preferred over the AME to assess response of broiler chickens to dietary GAA supplementation.

Periconceptional Folic Acid Supplementation Benefit to Development of Early Sensory-Motor Function through Increase DNA Methylation in Rat Offspring.

Periconceptional maternal folate levels may alter DNA methylation patterns and health outcomes in offspring. We hypothesized that maternal folic acid supplementation alters fetal neural development through DNA methylation in the fetal brain. Twenty-eight rats were randomly assigned to four groups: three groups of the female rats were fed folate-normal, folate-deficient or folate-supplemented diets from seven days before mating to delivery. In another group, folic acid supplementation diet short-period group was fed a folate-normal diet, except for 10 days (begin mating) when this group was fed a folate-supplemented diet. After delivery, the diets were changed to folate-normal diet for all four groups. The cliff avoidance and forelimb grip tests were used to assess sensory motor function of rat offspring. The results indicate that maternal folic acid supplementation improved the early development of sensory-motor function in offspring. Maternal folic acid supplementation increased the methylation potential, global DNA methylation (5-mC) and DNA methyltransferase expression and activity in the brains of the offspring. In conclusion, maternal folic acid supplementation increases DNA methylation pattern in offspring brain and improves the early development of sensory-motor function.

Rapid determination of ginkgolic acids in Ginkgo biloba kernels and leaves by direct analysis in real time-mass spectrometry.

A novel method based on direct analysis in real time integrated with mass spectrometry was established and applied into rapid determination of ginkgolic acids in Ginkgo biloba kernels and leaves. Instrument parameter settings were optimized to obtain the sensitive and accurate determination of ginkgolic acids. At the sample introduction speed of 0.2mm/s, high intensity of [M-H]- ions for ginkgolic acids were observed in the negative ion mode by utilization of high-purity helium gas at 450°C. Two microliters of methanol extract of G. biloba kernels or leaves dropped on the surface of Quick-Strip module was analyzed after solvent evaporated to dryness. A series of standard solutions of ginkgolic acid 13:0 in the range of 2-50mg/L were analyzed with a correlation coefficient r=0.9981 and relative standard deviation (n=5) from 12.5 to 13.7%. The limit of detection was 0.5mg/L. The results of direct analysis in real time-mass spectrometry were in agreement with those observed by thermochemolysis gas chromatography. The proposed method demonstrated significant potential in the application of the high-throughput screening and rapid analysis for ginkgolic acids in dietary supplements.

Hepatic Tmem30a Deficiency Causes Intrahepatic Cholestasis by Impairing Expression and Localization of Bile Salt Transporters

Mutations in ATP8B1 or ATP11C (members of P4-type ATPases) cause progressive familial intrahepatic cholestasis type 1 in human or intrahepatic cholestasis in mice. Transmembrane protein 30A (TMEM30A), a β-subunit, is essential for the function of ATP8B1 and ATP11C. However, its role in the etiology of cholestasis remains poorly understood. To investigate the function of TMEM30A in bile salt (BS) homeostasis, we developed Tmem30a liver-specific knockout (LKO) mice. Tmem30a LKO mice experienced hyperbilirubinemia, hypercholanemia, inflammatory infiltration, ductular proliferation, and liver fibrosis. The expression and membrane localization of ATP8B1 and ATP11C were significantly reduced in Tmem30a LKO mice, which correlated with the impaired expression and localization of BS transporters, such as OATP1A4, OATP1B2, NTCP, BSEP, and MRP2. The proteasome inhibitor bortezomib partially restored total protein levels of BS transporters but not the localization of BS transporters in the membrane. Furthermore, the expression of nuclear receptors, including FXRα, RXRα, HNF4α, LRH-1, and SHP, was also down-regulated. A cholic acid-supplemented diet exacerbated the liver damage in Tmem30a LKO mice. TMEM30A deficiency led to intrahepatic cholestasis in mice by impairing the expression and localization of BS transporters and the expression of related nuclear receptors. Therefore, TMEM30A may be a novel genetic determinant of intrahepatic cholestasis.

Expression and regulation of the neutral amino acid transporter B0AT1 in rat small intestine.

Absorption of neutral amino acids across the luminal membrane of intestinal enterocytes is mediated by the broad neutral amino acid transporter B0AT1 (SLC6A19). Its intestinal expression depends on co-expression of the membrane-anchored peptidase angiotensin converting enzyme 2 (ACE2) and is additionally enhanced by aminopeptidase N (CD13). We investigated in this study the expression of B0AT1 and its auxiliary peptidases as well as its transport function along the rat small intestine. Additionally, we tested its possible short- and long-term regulation by dietary proteins and amino acids. We showed by immunofluorescence that B0AT1, ACE2 and CD13 co-localize on the luminal membrane of small intestinal villi and by Western blotting that their protein expression increases in distal direction. Furthermore, we observed an elevated transport activity of the neutral amino acid L-isoleucine during the nocturnal active phase compared to the inactive one. Gastric emptying was delayed by intragastric application of an amino acid cocktail but we observed no acute dietary regulation of B0AT1 protein expression and L-isoleucine transport. Investigation of the chronic dietary regulation of B0AT1, ACE2 and CD13 by different diets revealed an increased B0AT1 protein expression under amino acid-supplemented diet in the proximal section but not in the distal one and for ACE2 protein expression a reverse localization of the effect. Dietary regulation for CD13 protein expression was not as distinct as for the two other proteins. Ring uptake experiments showed a tendency for increased L-isoleucine uptake under amino acid-supplemented diet and in vivo L-isoleucine absorption was more efficient under high protein and amino acid-supplemented diet. Additionally, plasma levels of branched-chain amino acids were elevated under high protein and amino acid diet. Taken together, our experiments did not reveal an acute amino acid-induced regulation of B0AT1 but revealed a chronic dietary adaptation mainly restricted to the proximal segment of the small intestine.