Arg Levels Research Articles

Effects of different forms of amino acid supplementation on the performance and intestinal barrier function of laying hens fed a low-protein diet

The aim of this study was to investigate the effects of low-protein diets and the sustained release of synthetic amino acids (AA) on the performance, intestinal barrier function and nitrogen excretion of laying hens. Two hundred eighty-eight 39-week-old Hyline brown laying hens of were randomly divided into 3 groups with 8 replicates per group. The crude protein level in the control group (CON) was 16%, the crude protein levels in the crystal AA supplement group (LCP-CAA) and microencapsulated AA group (LCP-MAA) were both 13%, and the AA levels in the LCP-CAA and LCP-MAA groups were consistent with that in the CON group. The experiment lasted 12 wk, and production performance was assessed weekly. The FCR and ADFI values were significantly greater for the LCP-CAA group than for the CON and LCP-MAA groups (P < 0.05). Two hours after feeding, His levels were significantly greater in the LCP-CAA group than in the LCP-MAA group (P < 0.05); 4 h after feeding, the contents of Met, Thr, Leu and Val were significantly greater in blood from the LCP-MAA group (P < 0.05); 6 h after feeding, Trp, Ile and Arg levels were significantly greater in the LCP-MAA group (P < 0.05). The chylase content significantly decreased in the duodenum of the LCP-CAA group (P < 0.05), and the chylase and trypsin were contents increased in the ileum of the LCP-MAA group (P < 0.05). In the LCP-MAA group, significantly increased mRNA expression levels of Occludin, ZO-1 in duodenum; Occludin, ZO-1, y+LAT1 in jejunum; and ZO-1 in ileum were detected at 8 and 12 weeks (P < 0.05). The fecal nitrogen content significantly decreased in the low protein diet group (P < 0.01). In conclusion, reducing dietary crude protein levels and supplementing with microencapsulated AAs can improve intestinal barrier function, promote digestive enzyme secretion, increase the expression of AA transporters, improve dietary protein utilization efficiency, and reduce nitrogen emission in laying hens.

Effect of Dietary Standardized Ileal Digestible Arginine to Lysine Ratio on Reproductive Performance, Plasma Biochemical Index, and Immunity of Gestating Sows.

The aim of this study was to determine the optimal SID Arg: Lys ratio for maximizing the reproductive performance, immunity and biochemical parameters of sows during gestation, the colostrum composition, and the performance of their offspring. A total of 174 multiparous sows were randomly allocated to five treatment groups varying in dietary SID Arg: Lys ratios (0.91, 1.02, 1.14, 1.25 and 1.38) through modification of the levels of Arg or alanine supplementation (the total level of nitrogen was the same in all treatments). The results showed that increasing the dietary SID Arg: Lys ratio increased the number of piglets born alive (p < 0.05, linear and quadratic). The number of stillborn piglets, the birth weight variation of born alive piglets, the birth interval (p < 0.05, linear and quadratic) and the number of mummies (p < 0.05, quadratic) reduced with increasing the dietary SID Arg: Lys ratio. Broken-line regression analysis indicated that the optimal SID Arg: Lys ratio requirement for gestating sows to maximize the number of piglets born alive was 1.25. The content of non-fat solid, total solid, protein, and energy in colostrum increased linearly and quadratically (p < 0.05) with increasing dietary SID Arg: Lys ratio. Additionally, when increasing the dietary SID Arg: Lys ratio, the concentration of IgA (p < 0.05, quadratic) and IgM (p < 0.05, linear and quadratic) of plasma increased at day 90 of gestation; IgG (p < 0.05, linear and quadratic) concentration increased at day 110 of gestation of sows. The dietary SID Arg: Lys ratio had an increasing effect (p < 0.05, linear and quadratic) on plasma insulin levels at day 90 of gestation. Furthermore, there were increases in plasma concentrations of nitric oxide and ornithine at day 110 of gestation, Arg at day 90 and 110 of gestation (p < 0.05, linear and quadratic) and total protein at day 110 of gestation (p < 0.05, linear) with increasing dietary SID Arg: Lys ratio. The results of our study indicated that increases in the dietary SID Arg: Lys ratio during gestation resulted in an increase in the number of piglets born alive, a decrease in birth intervals, and an improvement in immunity and colostrum composition. The optimal SID Arg: Lys ratio for gestating sows to maximize the number of piglets born alive was 1.25.

High-level L-Gln compromises intestinal amino acid utilization efficiency and inhibits protein synthesis by GCN2/eIF2α/ATF4 signaling pathway in piglets fed low-crude protein diets

Gln, one of the most abundant amino acids (AA) in the body, performs a diverse range of fundamental physiological functions. However, information about the role of dietary Gln on AA levels, transporters, protein synthesis, and underlying mechanisms in vivo is scarce. The present study aimed to explore the effects of low-crude protein diet inclusion with differential doses of L-Gln on intestinal AA levels, transporters, protein synthesis, and potential mechanisms in weaned piglets. A total of 128 healthy weaned piglets (Landrace × Yorkshire) were randomly allocated into four treatments with four replicates. Pigs in the four groups were fed a low-crude protein diet containing 0%, 1%, 2%, or 3% L-Gln for 28 d. L-Gln administration markedly (linear, P < 0.05) increased Ala, Arg, Asn, Asp, Glu, Gln, His, Ile, Lys, Met, Orn, Phe, Ser, Thr, Tyr, and Val levels and promoted trypsin activity in the jejunal content of piglets. Moreover, L-Gln treatment significantly enhanced concentrations of colonic Gln and Trp, and serum Thr (linear, P < 0.01), and quadratically increased serum Lys and Phe levels (P < 0.05), and decreased plasma Glu, Ile, and Leu levels (linear, P < 0.05). Further investigation revealed that L-Gln administration significantly upregulated Atp1a1, Slc1a5, Slc3a2, Slc6a14, Slc7a5, Slc7a7, and Slc38a1 relative expressions in the jejunum (linear, P < 0.05). Additionally, dietary supplementation with L-Gln enhanced protein abundance of general control nonderepressible 2 (GCN2, P = 0.010), phosphorylated eukaryotic initiation factor 2 subunit alpha (eIF2α, P < 0.001), and activating transcription factor 4 (ATF4) in the jejunum of piglets (P = 0.008). These results demonstrated for the first time that a low crude protein diet with high-level L-Gln inclusion exhibited side effects on piglets. Specifically, 2% and 3% L-Gln administration exceeded the intestinal utilization capacity and compromised the jejunal AA utilization efficiency, which is independent of digestive enzyme activities. A high level of L-Gln supplementation would inhibit protein synthesis by GCN2/eIF2α/ATF4 signaling in piglets fed low-protein diets, which, in turn, upregulates certain AA transporters to maintain AA homeostasis.

Human Umbilical Cord Mesenchymal Stem Cell-derived Exosomes Induce Macrophage M2 Polarization by Antagonizing LPS-mediated Stimulation of the NF-κB and STAT3 Pathways.

Many studies have documented the protective effects of regulating macrophage M1/M2 polarization in inflammatory diseases characterized by their imbalance state. In pathological diseases associated with inflammation, mesenchymal stem cells (MSCs) regulate macrophages, thereby having anti-inflammatory and tissue regenerative effects. Exosomes have been suggested as an alternative mechanism that underlies the paracrine function of MSCs. Thus, this study explored the anti-inflammatory impact of human umbilical cord MSCssecreted exosomes (hucMSCs-EX) by influencing macrophage polarization in normal and inflammatory environments in vitro. In this study, hucMSCs-conditioned medium (hucMSCs-CM) and hucMSCs- EX were used to treat RAW264.7 macrophages with or without LPS. The expressions of TNF- α, IL-10, IL-6, IL-1β, and Arg-1 were quantified by qPCR. The expressions of IL-6 and IL-10 were evaluated by ELISAs. Western blots (WB) were performed to observe the expressions of CD206, NF-κB P65, NF-κB p-p65, p-STAT3, STAT3, and NF-κB phosphorylation. The number of cells expressing CD206 and the fluorescence intensity were measured via flow cytometry (FC) and immunofluorescence staining. Cell propagation and migration were examined via MTT and transwell assays, respectively. The inhibition of LPS-induced inflammatory polarization by hucMSCs-EX or hucMSCs- CM led to increases in IL-10 and arginase (Arg) levels and decreases in those of IL-6 and TNF-α. Moreover, hucMSCs-EX enhanced the CD206 expression in RAW264.7 cells and accelerated the propagation and migration of LPS-induced cells. The suppressive impact of hucMSCs-EX on the LPS-induced phenotypic polarization of M1 macrophages was linked with the reduction of NF-κB signaling. They stimulated the transition of M2 macrophages by enhancing the activity of STAT3 in RAW264.7 cells. This study indicated that hucMSCs-EX enhances the macrophage transition into the M2 phenotype by inhibiting the NF-κB p65 axis and stimulating that of STAT3.

90 Evaluating water- and feed-based arginine supplementation on newly weaned pig growth and gastrointestinal health

Abstract The objective of this experiment was to assess the influence of arginine (Arg) supplementation in water and/or feed on the growth performance and gastrointestinal health of newly weaned pigs. Pigs (n = 240; PIC 337 Χ 1050, Genus, Henderson, TN) were randomly allocated into 80 mixed-sex test pens and subjected to a 2 Χ 4 factorial design, with two levels of Arg supplementation in water (0% or 8% stock, dosed through a 1:128 proportioner) for the first phase (d 0-7), and four dietary arginine levels (0.85, 0.95, 1.05, and 1.15 standardized ileal digestible (SID) Arg to Lysine (Lys) ratios for the first two phases (d 0-7 and 7-21). All treatments were provided a common diet (0.96 SID Arg:Lys) for the last phase d 21- 42. One pig per pen underwent a dual sugar absorption test of lactulose at 500 mg/kg and mannitol at 50 mg/kg of body weight (BW) via gastric tube on d 7 and 21, with blood plasma collected 4 h post-administration. The pig tested on d 7 was subsequently euthanized for intestinal tissue collection. Pen growth performance and feed disappearance were evaluated for 3 phases; d 0-7, 7-21, and 21-42. The statistical analysis utilized generalized linear mixed model methods (SAS 9.4, Cary, NC) with the main effects of SID Arg:Lys in the feed and Arg level within the water, and the interaction between the two. Pen was the experimental unit. Orthogonal contrasts were tested for the linear and quadratic effects of the diet. Growth performance during the 1st period exhibited variability, reflected by negative gain to feed ratio (G:F), caused by health challenges of Rotavirus A. Consequently, data were analyzed separately for each phase. Increasing dietary SID Arg:Lys caused a significant linear effect (improvement; P = 0.04) on final BW (18.47 and 21.90 kg, for 0.85 and 1.15 SID Arg:Lys, respectively). However, discernible differences between dietary treatments were not observed (P &amp;gt; 0.1). Similarly, a trend (P = 0.09) suggested a linear impact of dietary SID Arg:Lys on average daily gain during the third phase (d 21-42). In terms of intestinal absorption and morphology, Arg supplementation, whether through water or diet, had no effect on lactulose and mannitol absorption on both d 7 and d 21, nor did it impact villous height, crypt depth, or the villi:crypt ratio in ileum tissues collected on d 7 post-weaning. In conclusion, dietary supplementation with Arg increased final BW but had no clear impacts on intestinal health within the variables measured, potentially impacted by the rotavirus diagnosis in the first week post wean.

The correlation between vitamin D3 and arginine metabolism levels in newborns with amino acid metabolism disorders.

This study aimed to explore the correlation between vitamin D3 and arginine (Arg) metabolism indicators in newborns with amino acid metabolism disorders. Based on clinical data, 30 newborns with amino acid metabolism diseases admitted to Shijiazhuang Fourth Hospital from June 2021 to June 2022 were selected as the disease group, and 30 healthy newborns from the same period were selected as the healthy group. After enrollment, blood samples were collected to measure the levels of Arg, Glycine (Gly), and vitamin D3 levels. The levels of Arg metabolism indicators and vitamin D3 levels in the 2 groups and the correlation between vitamin D3 levels and Arg metabolism indicators in the affected group were analyzed. The Arg level in the diseased group was higher than that in the healthy group, whereas the Gly and vitamin D3 levels were lower than those in the healthy group (P < .05). There was a significant negative correlation between vitamin D3 and Arg levels in the affected group, and a significant positive correlation with Gly levels (P < .05). Newborns with amino acid metabolism disorders have abnormally high Arg levels, significantly reduced Gly levels, and significantly decreased vitamin D3 levels. The degree of decline was closely related to the levels of indicators of Arg metabolism. Vitamin D3 supplementation can improve the Arg metabolism status of newborns with amino acid metabolism disorders.

The Role of the L-Arginine-Nitric Oxide Molecular Pathway in Autosomal Dominant Polycystic Kidney Disease.

Recently, arginine has been proven to play an important role in ADPKD physiopathology. Arginine auxotrophy in ADPKD induces cell hyperproliferation, blocking the normal differentiation of renal tube cells and causing cyst formation. We explored the L-arginine (Arg)-nitric oxide (NO) molecular pathway in ADPKD, a multisystemic arginine auxotrophe disease. We developed a prospective case-control study that included a group of 62 ADPKD subjects with an estimated filtration rate over 60 mL/min/1.73 mp, 26 subjects with chronic kidney disease with an eGFR > 60 mL/min/1.73 mp, and a group of 37 healthy subjects. The laboratory determinations were the serum level of arginine, the enzymatic activity of arginase 2 and inducible nitric oxide synthase, the serum levels of the stable metabolites of nitric oxide (nitrate, direct nitrite, and total nitrite), and the endogenous inhibitors of nitric oxide synthesis (asymmetric dimethylarginine and symmetric dimethylarginine). In the ADPKD group, the levels of the arginine and nitric oxide metabolites were low, while the levels of the metabolization enzymes were higher compared to the control group. Statistical analysis of the correlations showed a positive association between the serum levels of Arg and the eGFR and a negative association between Arg and albuminuria. ADPKD is a metabolic kidney disease that is auxotrophic for arginine. Exploring arginine reprogramming and L-Arg-NO pathways could be an important element in the understanding of the pathogenesis and progression of ADPKD.

Inosine pretreatment of pregnant rats ameliorates maternal inflammation-mediated hypomyelination in pups via microglia polarization switch

Periventricular leukomalacia (PVL) is a neurological condition observed in premature infants, characterized by hypomyelination and activation of microglia. Maternal inflammation-induced brain injury in offspring significantly contributes to the development of PVL. Currently, there are no clinical pharmaceutical interventions available for pregnant women to prevent maternal inflammation-mediated brain injury in their offspring. Inosine has been shown to modulate the immune response in diverse stressful circumstances, such as injury, ischemia, and inflammation. The aim of this investigation was to examine the potential prophylactic impact of inosine on offspring PVL induced by maternal inflammation. This was accomplished by administering a 1 mg/ml inosine solution (40 ml daily) to pregnant Sprague-Dawley (SD) rats for 16 consecutive days prior to their intraperitoneal injection of lipopolysaccharide (350 µg/kg, once a day, for two days). The results showed that maternal inosine pretreatment significantly reversed the reduction in MBP and CNPase (myelin-related markers), CC-1 and Olig2 (oligodendrocyte-related markers) in their PVL pups (P7), suggesting that inosine administration during pregnancy could improve hypomyelination and enhance the differentiation of oligodendrocyte precursor cells (OPCs) in their PVL pups. Furthermore, the protective mechanism of inosine against PVL is closely associated with the activation and polarization of microglia. This is evidenced by a notable reduction in the quantity of IBA 1-positive microglia, a decrease in the level of CD86 (a marker for M1 microglia), an increase in the level of Arg 1 (a marker for M2 microglia), as well as a decrease in the level of pro-inflammatory factors TNF-α, IL-1β, and IL-6, and an increase in the level of anti-inflammatory factors IL-4 and IL-10 in the brain of PVL pups following maternal inosine pretreatment. Taken together, inosine pretreatment of pregnant rats can improve hypomyelination in their PVL offspring by triggering the M1/M2 switch of microglia.

Myofibrillar Protein Profile of The Breast Muscle in Turkeys as a Response to The Variable Ratio of Limiting Amino Acids in Feed

The effects of the different dietary levels of Arginine (Arg) in low- and high-methionine (Met) diets on the meat quality and myofibrillar protein profile of breast muscles from turkeys were determined. The experiment had a completely randomized 3 × 2 factorial design with three levels of Arg (90%, 100%, and 110%) relative to the dietary Met levels (30% or 45%). At 42 days of age, eight turkeys from each treatment were sacrificed; the meat pH value was measured at 48 h post-slaughter, and meat color was measured according to the CIE L*a*b* system. The SDS-PAGE method was performed to investigate the myofibrillar protein profile of the breast muscle. The analysis of variance showed a significant effect of the Arg or Met dietary levels on the color parameter b* and the profile of myofibrillar proteins in muscles. The results of the cluster analysis of the myofibrillar protein profile showed that, with a high level of Arg (i.e., 110%), the level of Met 35% or 45% was less important. It can be concluded that the increase in the share of Arg and Met in the diet of turkeys increases the content of some myofibrillar proteins (actinin, desmin, actin) and reduces degradation during the post-slaughter proteolysis of proteins that are considered tenderization indicators.

Impact of increasing dietary standardized ileal digestible arginine to lysine ratio from 0.85 to 1.15 and water-based arginine supplementation on growth performance and gut integrity of weaned pigs.

The objective of this experiment was to assess the influence of arginine (Arg) supplementation in water and/or feed on the growth performance and gastrointestinal health of newly weaned pigs. Two hundred and forty pigs (5.06kg; PIC, Hendersonville, TN) were randomly allocated into 80 mixed-sex pens (3 pigs/pen) and subjected to a 2 × 4 factorial design. Two levels of Arg were supplemented in water (0% or 8% stock, dosed through a 1:128 proportioner) for the first phase (days 0 to 7), and four dietary arginine levels (0.85, 0.95, 1.05, and 1.15) standardized ileal digestible (SID) Arg to Lysine (Lys) ratios for the first two phases (days 0 to 7 and 7 to 21). All treatments were provided a common diet (0.96 SID Arg:Lys) for the last phase days 21 to 42. One pig per pen underwent a dual sugar absorption test of lactulose at 500mg/kg and mannitol at 50mg/kg of body weight (BW) via gastric tube on days 7 and 21 postweaning, with blood plasma collected 4h later. The pig tested on day 7 was subsequently euthanized for intestinal tissue collection. Pen growth performance and feed disappearance were evaluated for 3 phases: days 0 to 7, 7 to 21, and 21 to 42 postweaning. The statistical analysis used linear models to examine the effects of SID Arg:Lys in the feed, Arg level in water, and their interactions, with pen as the experimental unit. Orthogonal contrasts were used to test the linear and quadratic effects of increasing SID Arg:Lys in the diet. Growth performance during the first period exhibited variability, reflected by negative gain-to-feed (G:F) ratios, caused by the enteric health challenge. Consequently, data were analyzed separately for each phase. Increasing dietary SID Arg:Lys caused a linear improvement (P = 0.04) in final BW (18.47 and 21.90kg, for 0.85 and 1.15 SID Arg:Lys, respectively). A trend (P = 0.09) suggested a linear impact of dietary SID Arg:Lys on average daily gain during days 21 to 42. Arg supplementation, whether administered through water or diet, did not affect lactulose and mannitol absorption on both days 7 and 21, nor did it alter histological measurements in the collected ileum tissues on day 7 postweaning. In conclusion, increasing dietary SID Arg:Lys increased final BW but had no clear impacts on intestinal health within the parameters measured, potentially impacted by the rotavirus diagnosis in the first week post-wean.

Effects of increasing dietary arginine supply during the three first weeks after weaning on pig growth performance, plasma amino acid concentrations, and health status.

A total of 425 weaned pigs (Exp. 1: 225 pigs [5.8 ± 0.9 kg]; Exp. 2: 200 pigs [6.1 ± 1.2 kg]) were used to determine the optimal dietary standardized ileal digestible (SID) arginine (Arg) level in early nursery diets based on growth and health responses. The basal diet in Exp.1 was formulated to meet SID Arg recommendation (0.66%; NRC, 2012) and in Exp. 2, SID Arg was set to simulate current industry practices for feeding nursery pigs (1.15 %). Basal diets were supplemented with 0.3%, 0.6%, 0.9%, and 1.2% of l-arginine to provide five levels of dietary SID Arg. Experimental diets were fed during phases I (days 0 to 7) and II (days 8 to 21) with common diets until market. Feed disappearance and body weight (BW) were measured on days 7, 14, 21, and 43. Final BW was recorded at first removal of pigs for market. Pen fecal score was assigned daily from days 0 to 21. Plasma immunoglobulin A (IgA) was determined on days 0, 7, and 14 and amino acids (AAs) concentration and plasma urea nitrogen (PUN) on days 0 and 14. Orthogonal polynomial contrasts were used to determine the linear and quadratic effects of dietary Arg. Optimal SID Arg was determined by fitting the data with piecewise regression, using growth performance as the primary response variable. In Exp. 1, dietary Arg linearly increased (P < 0.1) BW, average daily gain (ADG), and gain to feed ratio (G:F) ratio on day 21, as well as reduced (χ2 = 0.004) the percentage of pigs that lost weight (PLW) in week 1 by 29%. Dietary Arg resulted in linear improvement (P = 0.082) of ADG for the overall nursery period and quadratic improvement (P < 0.1) of final BW at marketing. In Exp. 2, dietary Arg linearly increased (P < 0.05) ADG and average daily feed intake (ADFI) in week 1, BW and ADFI (P < 0.1) on day 14, as well as reduced (χ2 ≤ 0.001) PLW in week 1. From days 0 to 21, G:F was improved quadratically (P < 0.1). Dietary Arg linearly increased (P < 0.1) ADG and BW on day 43. Dietary Arg supplementation decreased the incidence (χ2 < 0.05) of soft and watery feces during the first weeks after weaning and lower concentration of plasma IgA on days 7 and 14. Dietary Arg linearly and/or quadratically influenced plasma AA concentrations (P < 0.05), including an increase in Arg, Leu, Phe, Val, citrulline, ornithine, and PUN concentrations. Overall, weaned pigs exhibit optimal nursery growth performance and health when provided with dietary SID Arg ranging from 1.5% to 1.9%. This dietary range contributes to a reduction in the occurrence of fall-back pigs and improvements in final BW at marketing.

Fenugreek extract improves diabetes-induced endothelial dysfunction via the arginase 1 pathway.

Endothelial dysfunction (ED) is an initiating trigger and key factor in vascular complications, leading to disability and mortality in individuals with diabetes. The research concerning therapeutic interventions for ED has gained considerable interest. Fenugreek, a commonly used edible plant in dietary consumption, has attracted significant attention due to its management of diabetes and its associated complications. The research presented in this study examines the potential therapeutic benefits of fenugreek in treating ED and investigates the underlying mechanism associated with its effects. The analysis on fenugreek was performed using 70% ethanol extract, and its chemical composition was analyzed using ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). In total, we identified 49 compounds present in the fenugreek extract. These compounds encompass flavonoids, saponins, and phospholipids. Then, the models of ED in streptozotocin-induced diabetic mice and high glucose-induced isolated rat aortas were established for research. Through vascular function testing, it was observed that fenugreek extract effectively improved ED induced by diabetes or high glucose. By analyzing the protein expression of arginase 1 (Arg1), Arg activity, Arg1 immunohistochemistry, nitric oxide (NO) level, and the protein expression of endothelial nitric oxide synthase (eNOS), p38 mitogen-activated protein kinase (p38 MAPK), and p-p38 MAPK in aortas, this study revealed that the potential mechanism of fenugreek extract in anti-ED involves the downregulation of Arg1, leading to enhanced NO production. Furthermore, analysis of serum exosomes carrying Arg activity indicates that fenugreek may decrease the activity of Arg transported by serum exosomes, potentially preventing the increase in Arg levels triggered by the uptake of serum exosomes by vascular endothelial cells. In general, this investigation offers valuable observations regarding the curative impact of fenugreek extract on anti-ED in diabetes, revealing the involvement of the Arg1 pathway in its mechanism.

The effect of fructose exposure on amino acid metabolism among Chinese community residents and its possible multi-omics mechanisms

The consumption of fructose has increased dramaticly during the last few decades, inducing a great increase in the risk of intrahepatic lipid accumulation, hypertriglyceridemia, hyperuricemia and cancer. However, the underlying mechanism has not yet been fully elucidated. Amino acid metabolism may play an important role in the process of the diseases caused by fructose, but there is still a lack of corresponding evidence. In present study, we provide an evidence of how fructose affects amino acids metabolism in 1895 ordinary residents in Chinese community using UPLC-QqQMS based amino acid targeted metabolomics and the underlying mechanism of fructose exposure how interferes with amino acid metabolism related genes and acetylated modification of proteome in the liver of rats model. We found people with high fructose exposure had higher levels of Asa, EtN, Asp, and Glu, and lower levels of 1MHis, PEtN, Arg, Gln, GABA, Aad, Hyl and Cys. The further mechanism study displayed amino acid metabolic genes of Aspa, Cndp1, Dbt, Dmgdh, and toxic metabolites such as N-acetylethanolamines accumulation, interference of urea cycle, as well as acetylated modification of key enzymes in glutamine metabolic network and glutamine derived NEAAs synthesis pathway in liver may play important roles in fructose caused reprogramming in amino acid metabolism. This research provides novel insights of the mechanism of amino acid metabolic disorder caused by fructose and supplies new targets for clinical therapy.

The main nutritional components in colored rice grains

Colored rice grains contain rich nutrients. To investigate the differences in nutrient composition among different colored rice varieties, LC-MS non-targeted lipidomics and targeted amino acid profiling were used to study the lipid and amino acid composition in four colored rice grains. The study analyzed the relationship between amino acids, mineral elements, crude protein (CP), crude fat (CF), carotenoids (CAROT), vitamin C (ASA), ATP, total sugar (TS), soluble protein (SP), and metabolites. A total of 505 lipid metabolites were identified. The results depicted that YZ6H had the highest levels of Arg, Asn, Met, Orn, and Try. HN had the highest levels of GABA, His, Lys, Thr, Tyr, Val, and Tau, which were significantly higher than those in YZ6H, YJN818, and YCN1H. PC (16:0/16:1,16:1/18:2,18:1/14:1,18:1/14:0,18:1/18:1,18:0/18:1), TG (16:0/18:3/22:2,22:4/18:2/18:2,16:0/20:2/20:4), and PE(20:4e/15:0) were significantly negatively correlated with GABA, Leu, Lys, Thr, Tyr, Val, Gln, His, and Ile. TG(18:2/17:1/18:2,15:0/18:2/18:2,15:0/18:2/18:3,15:0/16:0/18:2,15:0/18:1/18:2,18:1/17:1/18:2,18:1/17:1/18:1,18:1/10:1/18:1) was significantly positively correlated with Pro and Tau. YZ6H had the highest levels of ASA, SP, TS, CAROT, and ATP content. PG(28:0/18:1), MLCL(14:2/16:0/16:0), Cer(t18:0/26:0+O,d18:2/16:0,t18:0/26:0), and LPC(16:0) TG(20:4e/18:3/20:1) were significantly negatively correlated with SP, CAROT. Hex1Cer(t18:1/24:0+O), MLCL(14:2/16:0/16:0), Cer(t18:0/26:0+O,d18:2/16:0,t18:0/26:0), LPC(16:0), TG(16:0/20:2/20:4,20:4e/18:3/20:1), and DG(18:1/14:0), which are key lipid metabolites involved in the synthesis of nutritional components in colored rice grains.

Growth performance, serum parameters, inflammatory responses, intestinal morphology and microbiota of weaned piglets fed 18% crude protein diets with different ratios of standardized ileal digestible isoleucine to lysine

The present study was to explore the Ile requirement of piglets fed 18% crude protein (CP) diets. Two hundred and fifty 28-day-old Duroc × Landrace × Yorkshire piglets (8.37 ± 1.92 kg) were randomly divided into 5 dietary treatments (10 piglets per replicate, 5 barrows and 5 gilts per replicate) with 45%, 50%, 55%, 60%, 65% standardized ileal digestible (SID) Ile-to-Lys ratios, and the SID Lys was formulated to 1.19%. The experimental design consisted of two phases (d 1 to 14 and d 15 to 28). Results showed that average daily gain (ADG) had a tendency to quadratically increase as the SID Ile-to-Lys ratio increased (P = 0.09), and the optimum SID Ile-to-Lys ratios required to maximize ADG were 48.33% and 54.63% for broken-line linear model and quadratic polynomial model, respectively. Different SID Ile-to-Lys ratios had no significant effects on average daily feed intake and gain-to-feed ratio. Dry matter (P < 0.01), CP (P = 0.01), ether extract (P = 0.04), gross energy (P < 0.01) and organic matter (P < 0.01) digestibility increased quadratically. Serum total cholesterol levels decreased linearly (P = 0.01) and quadratically (P < 0.01); aspartate aminotransferase (P < 0.01), interleukin-1β (P = 0.01), and tumor necrosis factor-α (P < 0.01) levels decreased quadratically; immunoglobulin G (P = 0.03) and immunoglobulin M (P = 0.01) concentrations increased quadratically. Serum Ser levels decreased linearly (P < 0.01) and quadratically (P = 0.01); Glu (P = 0.02), Arg (P = 0.05), and Thr (P = 0.03) levels decreased quadratically; Gly (P < 0.01) and Leu (P = 0.01) levels decreased linearly; Ile (P < 0.01) concentration increased linearly. Duodenal villus height (P < 0.01) and villus height to crypt depth ratio (P < 0.01) increased quadratically. The deficiency or excess of Ile decreased short chain fatty acid-producing bacteria abundance and increased pathogenic bacteria abundance. Overall, taking ADG as the effect index, the optimum SID Ile-to-Lys ratios of piglets offered 18% CP diets were 48.33% and 54.63% based on two different statistical models, respectively, and the deficiency or excess of lle negatively affected piglet growth rates and health status.

Mechanism(s)-of-Action for Arginine Therapy in Children with Sickle Cell Disease and Vaso-Occlusive Pain: Results of a Pharmacokinetics/Pharmacodynamics Study

Introduction: Low L-Arginine (Arg) bioavailability is associated with sickle cell disease (SCD) mortality and morbidity, including acute pain severity. Multiple phase 2 trials support the safety and efficacy of Arg therapy in children with SCD experiencing vaso-occlusive pain. Nitric oxide (NO) is a potent vasodilator depleted in SCD due to hemolysis. As the obligate substrate for NO production, Arg's mechanism-of-action is unknown but thought to be related in part to NO production. Kyotorphin (KTP) is an endogenous opioid analgesic composed of the amino acids tyrosine and Arg. Its nociceptive effects are through induction of Met-enkephalins release in the brain. Oral administration of Arg in wild-type mice increases the levels of KTP centrally. We have previously shown that Arg supplementation increases NO metabolites (NOx), improves mitochondrial function, decreases oxidative stress, and has opioid sparing effects in children with SCD. The association between IV Arg supplementation as a precursor to KTP in SCD is unknown and may represent a novel mechanism by which Arg decreases pain in patients with SCD. Objective: To analyze the results of a pharmacokinetics/pharmacodynamics study, evaluating the association between Arg supplementation and plasma levels of KTP and other potential mechanisms-of-action related to Arg in children with SCD and vaso-occlusive pain episodes (VOE). Methods: We conducted a pharmacokinetics/pharmacodynamics study of children aged 7-21 years with SCD hospitalized for VOE and randomized to treatment utilizing one of 3 dosing arms of Arg: 1) 100mg/kg IV three times/day, standard dose (n=4); 2) loading dose (200mg/kg) followed by standard dose (n=5); or 3) loading dose (200mg/kg) followed by continuous infusion (300mg/kg/day) until discharge (n=4). Plasma amino acids (arginine, ornithine, citrulline), global arginine bioavailability ratio (GABR; Arginine/[Citrulline+Ornithine]), KTP, NOx, exhaled NO, plasma arginase activity, and arginase-1 and -2 were measured. Mean±SD, paired t-tests, and Pearson correlation analyses between groups were performed where appropriate. Results: A total of 13 patients (age 13±3 years, 56% male, 88% Hb-SS, 85% on hydroxyurea) were enrolled. Statistically significant increases in plasma arginine levels (Fig1A), plasma KTP levels (Fig1B), and GABR from pre-dose (0 hour) to 1, 1.5, and 2 hours after arginine infusion were observed, with a return to baseline 8 hours after Arg administration. Arg and KTP levels peaked between 60-90 minutes after arginine infusion in all study arms (p&amp;lt;0.001, p=0.004 respectively). There were no significant differences in peak concentration across all study arm doses. Rise in KTP levels strongly correlated with changes in plasma Arg concentration ( r=0.72, p&amp;lt;0.0001). Plasma NOx levels significantly increased from pre-dose to Tmax (Tmax= 1-2 hours; mean percent change 156±313%, p=0.02), as did exhaled NO (mean percent change 31±27%, p=0.02). Arginase activity also increased significantly after IV Arg infusion peaking at approximately 90 minutes (mean percent change 137±183%, p= 0.03) with return to baseline at 8 hours, while there was no significant change in arginase-1 or -2 concentrations over time. Conclusion: IV Arg therapy increased plasma arginine concentration 2-5 times above baseline at presentation for VOE, and GABR also increased as expected in this pharmacokinetics study with both reaching a maximum concentration within 1 hour of infusion initiation in all dosing schemes. IV Arg supplementation increases plasma NOx levels suggesting a role for vasodilation as a potential mechanism-of-action during SCD-VOE. Exhaled NO also increased significantly, consistent with previous reports. Interestingly, IV Arg supplementation acutely increased arginase activity without a significant change in arginase concentration. Most notably, we report for the first time the impact of IV Arg therapy on plasma levels of KTP in SCD. As KTP is an endogenous opioid analgesic, this may represent a novel mechanism-of-action of Arg therapy that warrants further investigation. This observation may also have implications for the potential use of arginine supplementation in pain syndromes beyond sickle cell disease.

Knock-down of IGFBP2 ameliorates lung fibrosis and inflammation in rats with severe pneumonia through STAT3 pathway

Objective To observe the mechanism of IGFBP2 knock-down in improving lung fibrosis and inflammation through STAT3 pathway in rats with severe pneumonia. Materials and Methods First, SP rat model was established. Then rats were divided into the Control group, the SP group, the SP + Lv-vector shRNA group, the SP + Lv-IGFBP2 shRNA group, the SP + Lv-vector group, and the SP + Lv-IGFBP2 group. The mRNA and protein levels of IGFBP2, NOS, CD206 and Arg 1 were detected by RT-qPCR and Western blot. IHC was used to check the positive expression of IGFBP2 and MCP1. A fully automated blood gas analyzer was used to detected PaCO2, CO2 content, PaO2 and SaO2. HE and Masson staining were performed to observe the lung tissue injury and collagen deposition of rats in each group. ELISA assays were used to calculate the levels of inflammatory factors IL-1β, IL-6, TNF-α, IL-4, and IL-10. Flow cytometry was conducted to acquire the ratio of M1-type AMs and M2-type AMs. Results Compared with the Control group, IGFBP2, iNOS, CD206, and Arg1 mRNA and protein expression levels, IGFBP2 and MCP1 positive expressions, PaCO2, p-STAT3/STAT3, p-JAK2/JAK2, IL-1β, IL-6, and TNF-α levels, the number of AMs and neutrophils, the proportion of M1 type AMs and the expressions of α-SMA, Collagen-I, Collagen III, and Fibronectin were significantly increased in SP rats (p < 0.05), while PaCO2, CO2, and SaO2, IL-4 and IL-10 levels, and the proportion of M2 type AMs decreased (p < 0.05). However, the knockdown of IGFBP2 reversed the above index trends. Conclusion Knock-down of IGFBP2 ameliorated lung injury in SP rats, inhibited inflammation and pulmonary fibrosis, and promoted M2-type transformation of AMs by activating the STAT3 pathway.

Effects of L-arginine, guanidinoacetic acid and L-citrulline supplementation in reduced-protein diets on bone morphology and mineralization of laying hens.

The alterations in feed ingredients and the nutrient matrix to produce reduced-protein diets may affect bone morphology and mineralization in laying hens. This study was implemented to determine the effects of L-arginine (Arg), guanidinoacetic acid (GAA), and L-citrulline (Cit) supplementation to Arg-deficient reduced-protein diets on bone morphology, strength, and mineralization status of laying hens. Individually housed Hy-Line Brown laying hens were evenly distributed to five dietary treatments with 25 replicates per treatment from 20 to 40wk of age. Treatments consisted of a standard protein diet (17% crude protein, SP), a reduced-protein diet deficient in Arg (13% crude protein, RP), and RP supplemented with Arg (0.35% Arg, RP-Arg), GAA (0.46% GAA equivalent to 0.35% Arg, RP-GAA), or Cit (0.35% Cit equivalent to 0.35% Arg, RP-Cit) to reach the Arg level of SP diets. Birds fed the SP diet had similar bone weight, ash, length, width, Seedor index, breaking strength, and serum mineral concentration, but higher toe B level (P<0.001) compared to those fed the RP diet at wk 40. Birds fed the SP diet consumed more but also excreted more K and B compared to those fed the RP diet (P<0.01). Birds fed the SP diet had lower Cu digestibility (P=0.01) and higher B retention (P<0.01) compared to those offered the RP diet. Supplementation of Arg, GAA, and Cit to the RP diet increased relative femur weight and length (P<0.001). Citrulline supplementation also increased relative tibia and femur ash, and Zn digestibility (P<0.05). Supplementation of GAA to the RP diet decreased serum Ca, P, and Mg levels, decreased tibia Fe and Mg levels and toe Mg level, but increased Al, Fe, Zn, and Mn digestibility (P<0.05). The current findings demonstrated the capacity of laying hens to adapt to low mineral intake by increasing mineral utilization. Overall, bone morphology and breaking strength, and serum mineral level in laying hens were not influenced by dietary CP levels. Dietary Arg, GAA, or Cit supplementation were effective in improving bone morphology and mineralization in laying hens fed Arg-deficient RP diets.

A ratiometric fluorescent dye for detection of Lys and Arg and its bioimaging in live cells and zebrafish larvae.

A ratiometric and pH-sensitive fluorescent dye named IDE was applied to the detection of argine and lysine from common amino acids and exploited to monitor the Lys and Arg levels in living cells and zebrafish larvae successfully. IDE will be a useful fluorescence indicator of pH changes by Lys and Arg.

Methionine and total homocysteine in hypertensive patients with renal excretory dysfunction.

The role of the kidneys in the metabolism and homeostasis of sulfur-containing amino acids is great, so the levels of methionine (Met), total homocysteine (tHcy) and their ratios can be of diagnostic value in chronic kidney disease (CKD), in a course of the arterial hypertension (AH). The aim of the study was to evaluate the Met/tHcy ratio in hypertensive patients with CKD. We used blood plasma of 76 patients aged 40-75 years with AH and the excretory dysfunction of the kidneys; subgroups: 1 - with proteinuria (n=37); 2 - without proteinuria with glomerular filtration rate (GFR) < 90 ml/min/1.73 m2 (n=39) and comparison group 3 - patients with AH without renal excretory dysfunction (n=28). Significantly lower Met levels were in subgroup 1. THcy levels were higher in subgroups 1 and 2 than in group 3. The Met/tHcy ratio revealed differences in subgroups 1 and 2 vs group 3. No differences were found in Arg and Lys levels. Positive correlations of the Met/tHcy ratio with the number of erythrocytes, but not with the level of hemoglobin, were revealed. In the ROC analysis, the cut-off points for the Met/tHcy ratio compared to group 3 were 3.08 for subgroup 1 and 3.36 for subgroup 2. With the progression of CKD, there is an increase in the levels of tHcy in the blood, and a decrease in the content of Met. A decrease in GFR, especially in a case with proteinuria, is accompanied by a decrease in the level of Met. The Met/tHcy ratio above 3.36 can be considered as the minimum of the balance between these sulfur-containing amino acids contents in a blood necessary for hypertensive patients with CKD.