Decoding the nexus:branched-chain amino acids and their connection with sleep,circadian rhythms,and cardiometabolic health

The sleep-wake cycle stands as an integrative process essential for sustaining optimal brain function and,either directly or indirectly,overall body health,encompassing metabolic and cardiovascular well-being.Given the heightened metabolic activity of the brain,there exists a considerable demand for nutrients in comparison to other organs.Among these,the branched-chain amino acids,comprising leucine,isoleucine,and valine,display distinctive significance,from their contribution to protein structure to their involvement in overall metabolism,especially in cerebral processes.Among the first amino acids that are released into circulation post-food intake,branched-chain amino acids assume a pivotal role in the regulation of protein synthesis,modulating insulin secretion and the amino acid sensing pathway of target of rapamycin.Branched-chain amino acids are key players in influencing the brain's uptake of monoamine precursors,competing for a shared transporter.Beyond their involvement in protein synthesis,these amino acids contribute to the metabolic cycles ofγ-aminobutyric acid and glutamate,as well as energy metabolism.Notably,they impact GABAergic neurons and the excitation/inhibition balance.The rhythmicity of branchedchain amino acids in plasma concentrations,observed over a 24-hour cycle and conserved in rodent models,is under circadian clock control.The mechanisms underlying those rhythms and the physiological consequences of their disruption are not fully understood.Disturbed sleep,obesity,diabetes,and cardiovascular diseases can elevate branched-chain amino acid concentrations or modify their oscillatory dynamics.The mechanisms driving these effects are currently the focal point of ongoing research efforts,since normalizing branched-chain amino acid levels has the ability to alleviate the severity of these pathologies.In this context,the Drosophila model,though underutilized,holds promise in shedding new light on these mechanisms.Initial findings indicate its potential to introduce novel concepts,particularly in elucidating the intricate connections between the circadian clock,sleep/wake,and metabolism.Consequently,the use and transport of branched-chain amino acids emerge as critical components and orchestrators in the web of interactions across multiple organs throughout the sleep/wake cycle.They could represent one of the so far elusive mechanisms connecting sleep patterns to metabolic and cardiovascular health,paving the way for potential therapeutic interventions.

Abomasal infusion of branched-chain amino acids or branched-chain keto-acids alter lactation performance and liver triglycerides in fresh cows

Background Dairy cows are at high risk of fatty liver disease in early lactation,but current preventative measures are not always effective.Cows with fatty liver have lower circulating branched-chain amino acid(BCAA)concentra-tions whereas cows with high circulating BCAA levels have low liver triglyceride(TG).Our objective was to determine the impact of BCAA and their corresponding ketoacids(branched-chain ketoacids,BCKA)on production performance and liver TG accumulation in Holstein cows in the first 3 weeks postpartum.Methods Thirty-six multiparous Holstein cows were used in a randomized block design experiment.Cows were abomasally infused for the first 21 d postpartum with solutions of 1)saline(CON,n=12);2)BCA(67 g valine,50 g leu-cine,and 34 g isoleucine,n=12);and 3)BCK(77 g 2-ketovaline calcium salt,57 g 2-ketoleucine calcium salt,and 39 g 2-ketoisoleucine calcium salt,n=12).All cows received the same diet.Treatment effects were determined using PROC GLIMMIX in SAS.Results No differences were detected for body weight,body condition score,or dry matter intake averaged over the first 21 d postpartum.Cows receiving BCK had significantly lower liver TG concentrations compared to CON(6.60%vs.4.77%,standard error of the mean(SEM)0.49)during the first 3 weeks of lactation.Infusion of BCA increased milk yield(39.5 vs.35.3 kg/d,SEM 1.8),milk fat yield(2.10 vs.1.69 kg/d,SEM 0.08),and lactose yield(2.11 vs.1.67 kg/d,SEM 0.07)compared with CON.Compared to CON,cows receiving BCA had lower plasma glucose(55.0 vs.59.2 mg/dL,SEM 0.86)but higherβ-hydroxybutyrate(9.17 vs.6.00 mg/dL,SEM 0.80).Conclusions Overall,BCAA supplementation in this study improved milk production,whereas BCKA supplementa-tion reduced TG accumulation in the liver of fresh cows.

Impairment of innate immune responses in cirrhotic patients and treatment by branched-chain amino acids

It has been reported that host defense responses, such as phagocytic function of neutrophils and natural killer (NK) cell activity of lymphocytes, are impaired in cirrhotic patients. This review will concentrate on the impairment of innate immune responses in decompensated cirrhotic patients and the effect of the treatment by branched-chain amino acids (BCAA) on innate immune responses. We already reported that phagocytic function of neutrophils was significantly improved by 3-mo BCAA supplementation. In addition, the changes of NK activity were also significant at 3 mo of supplementation compared with before supplementation. Also, Fisher’s ratios were reported to be significantly increased at 3 mo of BCAA supplementation compared with those before oral supplementation. Therefore, administration of BCAA could reduce the risk of bacterial and viral infection in patients with decompensated cirrhosis by restoring impaired innate immune responses of the host. In addition, it was also revealed that BCAA oral supplementation could reduce the risk of development of hepatocellular carcinoma in cirrhotic patients. The mechanisms of the effects will also be discussed in this review article.

Effect of branched-chain amino acids in patients receiving intervention for hepatocellular carcinoma

AIM: To investigate the usefulness of branched-chain amino acids (BCAA) before transarterial chemoembolization (TACE) or radiofrequency ablation (RFA).

Branched-chain amino acids to tyrosine ratio value as a potential prognostic factor for hepatocellular carcinoma

The prognosis of hepatocellular carcinoma(HCC) depends on tumor extension as well as hepatic function.Hepatic functional reserve is recognized as a factor affecting survival in the treatment of HCC;the ChildPugh classif ication system is the most extensively used method for assessing hepatic functional reserve in patients with chronic liver disease,using serum albumin level to achieve accurate assessment of the status of protein metabolism.However,insuff icient attention has been given to the status of amino acid(AA) metabolism in chronic liver disease and HCC.Fischer's ratio is the molar ratio of branched-chain AAs(BCAAs:leucine,valine,isoleucine) to aromatic AAs(phenylalanine,tyrosine) and is important for assessing liver metabolism,hepatic functional reserve and the severity of liver dysfunction.Although this ratio is diff icult to determine in clinical situations,BCAAs/tyrosine molar concentration ratio(BTR) has been proposed as a simpler substitute.BTR correlates with various liver function examinations,including markers of hepatic f ibrosis,hepatic blood flow and hepatocyte function,and can thus be considered as reflecting the degree of hepatic impairment.This manuscript examines the literature to clarify whether BTR can serve as a prognostic factor for treatment of HCC.

Branched-chain amino acids supplementation has beneficial effects on the progression of liver cirrhosis:A meta-analysis

BACKGROUND Liver cirrhosis(LC)is currently the 11^(th) most common cause of death and 15^(th) cause of morbidity globally.The treatment of LC is mainly aimed at etiological intervention,lifestyle intervention,prevention and treatment of complications and nutritional treatment.Nutritional treatment of LC mainly includes increasing dietary intake,food intake time and branched-chain amino acids(BCAAs).Despite the recommendation of BCAAs in some guidelines,adverse effects have been reported in studies so the efficacy and safety of BCAAs remain controversial.Currently,BCAAs have been widely used in chronic liver disease,while the summary of the effect of BCAAs on long-term prognosis is rare.AIM To determine the effects of BCAAs in patients with LC.METHODS The PubMed,Cochrane Library,Embase and Web of Science databases were searched.The retrieval deadline was 1 October 2021 and there were no language restrictions set in the retrieval.The study was performed in strict accordance with the inclusion and exclusion criteria.Nine studies were finally included.The primary outcome was complications of LC.The secondary outcomes were nutritional status and liver function.This meta-analysis used the Review Manager,version 5 statistical package(Cochrane Collaboration,Oxford,England)for analysis.RESULTS The analysis included nine studies that consisted of 1080 patients(554 in the BCAA groups and526 in the control groups).The nine studies were randomized control trials(RCTs).The quality of the studies was assessed using the risk of bias method recommended by the Cochrane Collaboration.BCAAs reduced the rate of complications in LC patients[Risk ratio:0.70,95%confidence interval(CI):0.56-0.88,P=0.002]and improved patients’albumin levels[std mean difference SMD:0.26,95%CI:0.12-0.40,P=0.0002].Meanwhile,BCAAs significantly ameliorated the levels of alanine transaminase(SMD:-2.03,95%CI:-2.52 to-1.53,P<0.00001)and aspartate aminotransferase(SMD:-1.8,95%CI:-2.14 to-1.46,P<0.00001).Meanwhile,glucose in the LC was significantly increased in BCAA-treated patients(MD:13.04,95%CI:6.81-19.89,P=0.0002).CONCLUSION BCAAs reduce the incidence of complications in patients with LC and ameliorate nutritional status.

Dietary branched-chain amino acids modulate the dynamics of calcium absorption and reabsorption in proteinrestricted pigs

Background:Very low-protein(VLP)diets negatively impact calcium(Ca)metabolism and absorption.The objective of this study was to investigate the effect of supplemental branched-chain amino acids(BCAA)and limiting amino acids(LAA)on Ca digestibility,absorption and reabsorption in pigs fed with VLP diets.Forty-eight piglets were assigned to six treatments:positive control(PC),negative control(NC),and NC containing LAA 25%,LAA 50%,LAA+BCAA 25%(LB25)and LAA+BCAA 50%(LB50)more than recommendations.Results:Relative to PC or NC,LB25 and LB50 had higher digestibility of Ca and plasma Ca and phosphorus(P),but lower plasma vitamin D3.LB50 tended to increase vitamin D receptor transcript and protein in the gut,but decreased mRNA or protein abundance of parathyroid hormone 1 receptor(PTH1R),calbindin 1(CALB1),cytochrome P450 family 27 subfamily B member 1 and occludin in small intestine.LB50 increased the transcript of cytochrome P450 family 24 subfamily A member 1 and PTH1R but decreased the transcript of transient receptor potential cation channel subfamily V member 5,CALB1 and solute carrier family 17 member 4 in kidney.Conclusion:Overall,BCAA increased Ca digestibility through regulating the transcellular and paracellular Ca absorption in the gut and reabsorption in kidney during protein restriction.

Effects of Branched-chain Amino Acids on Nutritional Metabolism and Pharmacoeconomics in Patients with Severe Abdominal Trauma

Objective:To observe the influences of branched-chain amino acids(BCAAs)on nutrition metabolism and prognosis of patients with severe abdominal trauma;at the same time,to analyze and evaluate the pharmacoeconomics of it.Methods:A total of 75 severe abdominal trauma patients were recruited from June 2016 to December 2017 and randomly divided into control group and observation group.After surgery and basic treatment,parenteral nutrition support therapy with iso-nitrogen and iso-calorie of both groups was administered.Meanwhile,an equivalent of 8.5%(18AA-Ⅱ)and 10%(20AA)compound AA injection was administrated to the control and observation groups,respectively.The nitrogen balance,serum protein level and plasma amino spectrum of the patients were observed before and after treatment.Besides,the hospital stay,survival rate,complications,adverse reactions and hospitalization costs were also compared.Results:After a 7-day course treatment,the nitrogen balance level of the two groups was significantly improved,but no significant difference was found between them.In addition,the serum protein level and plasma amino spectrum of the two groups was generally improved when compared to before treatment.Compared with the control group,the level of albumin and transferrin in the observation group was improved significantly after treatment,while no difference in plasma amino spectrum was found between the two groups.Moreover,the cost analysis showed remarkably reduced hospitalization costs in the observation group.Conclusion:To a certain degree,BCAAs could improve the nutritional metabolism and prognosis of patients with severe abdominal trauma,and have good cost-effectiveness.

Functional microbiomics reveals branched-chain amino acids depletion for alleviation of insulin resistance by berberine

Increased circulating branched-chain amino acids(BCAAs)have been involved in the pathogenesis of obesity and insulin resistance.However,evidence relating berberine(BBR),gut microbiota,BCAAs,and insulin resis⁃tance is limited.Here,we showed that BBR could effectively rectify steatohepatitis and glucose intolerance in high-fat diet(HFD)-fed mice.BBR reorganized gut microbiota populations under both the normal chow diet(NCD)and HFD.Particu⁃larly,BBR noticeably decreased the relative abundance of BCAA-producing bacteria,including order Clostridiales;fami⁃lies Streptococcaceae,Clostridiaceae,and Prevotellaceae;and genera Streptococcus and Prevotella.Compared with the HFD group,predictive metagenomics indicated a reduction in the proportion of gut microbiota genes involved in BCAA biosynthesis but the enrichment genes for BCAA degradation and transport by BBR treatment.Accordingly,the elevated serum BCAAs of HFD group were significantly decreased by BBR.Furthermore,the Western blotting results implied that BBR could promote the BCAA catabolism in the liver and epididymal white adipose tissues of HFD-fed mice by acti⁃vation of the multienzyme branched-chain α-ketoacid dehydrogenase complex,whereas by inhibition of the phosphoryla⁃tion state of BCKDHA(E1α subunit)and branched-chain α-ketoacid dehydrogenase kinase.The ex vivo assay further confirmed that BBR could increase BCAA catabolism in both AML12 hepatocytes and 3T3-L1 adipocytes.Finally,data from healthy subjects and diabetics confirmed that BBR could improve glycemic control and modulate circulating BCAAs.Besides,functional microbiomics integrated high-throughput microbial genomics,metabolomics and molecular biotechnology has also been successfully applied to reveal the anti-obesity mechanism of hydroxysafflor yellow A.

Branched-chain amino acids in liver diseases

Branched chain amino acids(BCAAs)have been shown to affect gene expression,protein metabolism,apoptosis and regeneration of hepatocytes,and insulin resistance.They have also been shown to inhibit the proliferation of liver cancer cells in vitro,and are essential for lymphocyte proliferation and dendritic cell maturation.In patients with advanced chronic liver disease,BCAA concentrations are low,whereas the concentrations of aromatic amino acids such as phenylalanine and tyrosine are high,conditions that may be closely associated with hepatic encephalopathy and the prognosis of these patients.Based on these basic observations,patients with advanced chronic liver disease have been treated clinically with BCAA-rich medicines,with positive effects.

Synergistic renoprotective effect of a compiled branched-chain amino acids and Cymbopogon schoenanthus extract against experimentally induced oxido-nitrosative renal insult

Objective: To better investigate the protective role of branched-chain amino acids(BCAAs)and Cymbopogon schoenanthus(CS) extract against the potassium dichromate(PDC)-induced oxido-nitrosative nephrotoxic insult in the experimental rat model. Methods: Thirty male rats were randomly divided into five equal groups: The 1 st group served as control; the 2^(nd)was injected with a single dose of PDC(15 mg/kg b.w i.p.);the 3^(rd), 4^(th), and 5^(th) groups were respectively treated with BCAAs, CS, and their combination for 15 d prior to induction of renal insult via PDC single dose(15 mg/kg b.w s.c.). The experimental period was terminated in all groups 2 d after induction of renal insult. The harvested kdney samples were divided for biochemical assays and histological examination. Results: The PDC-induced nephrotoxic effect caused a depletion of renal oxidative scavengers glutathione, superoxide dismutase with consequent lipo-oxidative cellular membrane deterioration manifested by a rise in malonaldehyde, oxidized glutathione, myeloperoxidase and the concomitant increase in inflammatory response elements tumor necrosis factor α, nitric oxide, and interleukin 1 β.Moreover, the comet assay and increased 8-hydroxy-2-deoxyguanosine proved an accelerated apoptotic DNA fragmentation. These local renal changes were met with global altered blood biochemistry. The BCAAs and CS or their compiled administration showed an ameliorative effect against PDC-induced nephrotoxic in a synergistic pattern. Conclusions: Both BCAAs and CS or their combined administration afford potential competitors against renal insult induced by polyvalent anion pollutants in experimentally studied animals model. As a route for novel drug discovery, further investigation should be attempted to optimize their augmenting reno-protecting potential.

Herbicide-Resistant Mutations in Acetolactate Synthase Can Reduce Feedback Inhibition and Lead to Accumulation of Branched-Chain Amino Acids

The branched-chain amino acids (BCAAs) valine, leucine and isoleucine are essential amino acids that are critical for animal growth and development. Animals need to obtain BCAAs from their diet because they cannot synthesize them. Plants are the ultimate source of these amino acids. Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of BCAAs. The metabolic control of BCAA biosynthesis involves allosteric regulation of ALS by the end-products of the pathway, i.e., valine, leucine and isoleucine. ALS holoenzyme seems to consist of two large catalytic subunits and two small regulatory subunits. In a previous study, using homologous recombination dependent gene targeting we created rice plants in which W548Land S627I mutations were induced into the endogenous gene encoding the ALS catalytic subunit. These two amino acid substitutions conferred hypertolerance to the ALS-inhibiting herbicide bispyripac-sodium. In this study, we revealed that feedback regulation by valine and leucine was reduced by these two amino acid substitutions. Furthermore, in leaves and seeds of ALS mutants with W548Land/or S627I substitution, a 2- to 3-fold increase in BCAAs was detected. Our results suggest that the ALS catalytic subunit is also involved in feedback regulation of ALS, and that judicious modification of the regulatory and catalytic subunits of ALS-coding genes by gene targeting can lead to the efficient accumulation of BCAA in plants.

Effects of L-carnitine and Branched-Chain Amino Acids on Energy Metabolism,Body Composition,and Delayed-Onset Muscle Soreness after Exercise in Healthy Subjects

Background Specific nutrients combined with exercise may prevent a loss of lean body mass and consequently prevent sarcopenia.We examined the effects of 2 weeks of supplementation with L-carnitine and a single dose of branched-chain amino acids(BCAAs)on energy metabolism and body composition before and after exercise and on post-exercise muscle soreness in healthy subjects.Methods A total of 12 young,untrained,healthy women were randomly assigned to two groups who received either 2 weeks of supplementation with L-carnitine(1000 mg/day)and a single dose of BCAAs plus 200 kcal energy(LCAR+BCAA group,n=6)or 200 kcal energy alone(control group,n=6)before 60 min of exercise.Energy substrate metabolism and body composition by bio-impedance were examined before and after exercise.Delayed-onset muscle soreness(DOMS)was evaluated by using a visual analogue scale for 3 days after exercise.Results Serum concentrations of free and acyl-L-carnitine in the LCAR+BCAA group were not significantly different from those in the control group.There were no significant differences in body composition before or after exercise between the two groups.Although serum free fatty acid levels 120 min after the oral ingestion of BCAAs were significantly lower in the LCAR+BCAA group than in the control group,there were no significant differences in serum free fatty acid levels immediately after and 60 min after exercise.DOMS for 3 days after exercise was lower in the LCAR+BCAA group than in the control group,but this difference was not significant.Conclusion Continuous supplementation with L-carnitine combined with a dose of BCAAs in young,untrained,healthy subjects caused a blunted response of stimulated lipolysis after exercise.

Heterogeneously elevated branched-chain/aromatic amino acids among new-onset type-2 diabetes mellitus patients are potentially skewed diabetes predictors

BACKGROUND The lack of specific predictors for type-2 diabetes mellitus(T2DM)severely impacts early intervention/prevention efforts.Elevated branched-chain amino acids(BCAAs:Isoleucine,leucine,valine)and aromatic amino acids(AAAs:Tyrosine,tryptophan,phenylalanine)show high sensitivity and specificity in predicting diabetes in animals and predict T2DM 10-19 years before T2DM onset in clinical studies.However,improvement is needed to support its clinical utility.AIM To evaluate the effects of body mass index(BMI)and sex on BCAAs/AAAs in new-onset T2DM individuals with varying body weight.METHODS Ninety-seven new-onset T2DM patients(<12 mo)differing in BMI[normal weight(NW),n=33,BMI=22.23±1.60;overweight,n=42,BMI=25.9±1.07;obesity(OB),n=22,BMI=31.23±2.31]from the First People’s Hospital of Yunnan Province,Kunming,China,were studied.One-way and 2-way ANOVAs were conducted to determine the effects of BMI and sex on BCAAs/AAAs.RESULTS Fasting serum AAAs,BCAAs,glutamate,and alanine were greater and high-density lipoprotein(HDL)was lower(P<0.05,each)in OB-T2DM patients than in NW-T2DM patients,especially in male OB-T2DM patients.Arginine,histidine,leucine,methionine,and lysine were greater in male patients than in female patients.Moreover,histidine,alanine,glutamate,lysine,valine,methionine,leucine,isoleucine,tyrosine,phenylalanine,and tryptophan were significantly correlated with abdominal adiposity,body weight and BMI,whereas isoleucine,leucine and phenylalanine were negatively correlated with HDL.CONCLUSION Heterogeneously elevated amino acids,especially BCAAs/AAAs,across new-onset T2DM patients in differing BMI categories revealed a potentially skewed prediction of T2DM development.The higher BCAA/AAA levels in obese T2DM patients would support T2DM prediction in obese individuals,whereas the lower levels of BCAAs/AAAs in NW-T2DM individuals may underestimate T2DM risk in NW individuals.This potentially skewed T2DM prediction should be considered when BCAAs/AAAs are to be used as the T2DM predictor.

Gut microbiota induced abnormal amino acids and their correlation with diabetic retinopathy

AIM:To explore the correlation of gut microbiota and the metabolites with the progression of diabetic retinopathy(DR)and provide a novel strategy to elucidate the pathological mechanism of DR.METHODS:The fecal samples from 32 type 2 diabetes patients with proliferative retinopathy(PDR),23 with nonproliferative retinopathy(NPDR),27 without retinopathy(DM),and 29 from the sex-,age-and BMI-matched healthy controls(29 HC)were analyzed by 16S rDNA gene sequencing.Sixty fecal samples from PDR,DM,and HC groups were assayed by untargeted metabolomics.Fecal metabolites were measured using liquid chromatographymass spectrometry(LC-MS)analysis.Associations between gut microbiota and fecal metabolites were analyzed.RESULTS:A cluster of 2 microbiome and 12 metabolites accompanied with the severity of DR,and the close correlation of the disease progression with PDR-related microbiome and metabolites were found.To be specific,the structure of gut microbiota differed in four groups.Diversity and richness of gut microbiota were significantly lower in PDR and NPDR groups,than those in DM and HC groups.A cluster of microbiome enriched in PDR group,including Pseudomonas,Ruminococcaceae-UCG-002,Ruminococcaceae-UCG-005,Christensenellaceae-R-7,was observed.Functional analysis showed that the glucose and nicotinate degradations were significantly higher in PDR group than those in HC group.Arginine,serine,ornithine,and arachidonic acid were significantly enriched in PDR group,while proline was enriched in HC group.Functional analysis illustrated that arginine biosynthesis,lysine degradation,histidine catabolism,central carbon catabolism in cancer,D-arginine and D-ornithine catabolism were elevated in PDR group.Correlation analysis revealed that Ruminococcaceae-UCG-002 and Christensenellaceae-R-7 were positively associated with L-arginine,ornithine levels in fecal samples.CONCLUSION:This study elaborates the different microbiota structure in the gut from four groups.The relative abundance of Ruminococcaceae-UCG-002 and Parabacteroides are associated with the severity of DR.Amino acid and fatty acid catabolism is especially disordered in PDR group.This may help provide a novel diagnostic parameter for DR,especially PDR.

Effect of different drying methods on the amino acids,α-dicarbonyls and volatile compounds of rape bee pollen

The significant demand for high quality food has motivated us to adopt appropriate processing methods to improve the food nutritional quality and flavors.In this study,the effects of five drying methods,namely,pulsed vacuum drying(PVD),freeze drying(FD),infrared drying(IRD),hot-air drying(HAD)and sun drying(SD)on free amino acids(FAAs),α-dicarbonyl compounds(α-DCs)and volatile compounds(VOCs)in rape bee pollen(RBP)were determined.The results showed that FD significantly released the essential amino acids(EAAs)compared with fresh samples while SD caused the highest loss.Glucosone was the dominantα-DCs in RBP and the highest loss was observed after PVD.Aldehydes were the dominant volatiles of RBP and SD samples contained more new volatile substances(especially aldehydes)than the other four drying methods.Comprehensively,FD and PVD would be potential methods to effectively reduce the quality deterioration of RBP in the drying process.

Standardized ileal digestibility of amino acids in soybean meal fed to non-pregnant and pregnant sows

Background Two studies were designed to determine standard ileal crude protein(CP)and amino acid(AA)digestibility of soybean meal(SBM)from different origins fed to non-pregnant and pregnant sows.Seven solvent-extracted SBMs from soybeans produced in the USA,Brazil,and China were selected.In Exp.1,eight different diets were created:a nitrogen(N)-free diet and 7 experimental diets containing SBM from different origins as the only N source.Eight non-pregnant,multiparous sows were arranged in an 8×8 Latin square design(8 periods and 8 diets).In Exp.2,the diet formula was the same as in Exp.1.Eight gestating sows(parity 3)were assigned to 4 different diets in a replicated 4×3 Youden square design(three periods and four diets)in mid-gestation and again in late-gestation stages.Results When fed to non-pregnant and late-gestating sows,the standardized ileal digestibility(SID)of CP and most AAs from different SBM were not significantly different(P>0.05).When fed to mid-gestating sows,the SID values for Arg,His,Lys,Phe,Cys,Gly,Ser,and Tyr in SBM 1 were lower than in SBM 4 and 5(P<0.05),whereas SID for Leu from SBM 5 was higher than in SBM 1 and 4(P<0.05).SID values for Ile,Ala,and Asp from SBM 4 were lower than in SBM 1 and 5(P<0.05).Sows had significantly greater SID values for Lys,Ala,and Asp during mid-gestation when compared with late-gestation stages(P<0.05).Mid-gestating sows had greater SID value for Val and lower SID value for Tyr when compared with non-pregnant and late-gestating sows(P<0.01),whereas non-pregnant sows had significantly greater SID value for Met when compared with gestating sows(P<0.01).Conclusions When fed to mid-gestating sows,the SID values for most AAs varied among SBM samples.The SID values for Lys,Met,Val,Ala,Asp,and Tyr in SBM were affected by sow gestation stages.Our findings provide a cornerstone for accurate SBM use in sow diets.

Targeting harmful effects of non-excitatory amino acids as an alternative therapeutic strategy to reduce ischemic damage

The involvement of the excitatory amino acids glutamate and aspartate in ce rebral ischemia and excitotoxicity is well-documented.Nevertheless,the role of non-excitatory amino acids in brain damage following a stroke or brain trauma remains largely understudied.The release of amino acids by necrotic cells in the ischemic core may contribute to the expansion of the penumbra.Our findings indicated that the reversible loss of field excitato ry postsynaptic potentials caused by transient hypoxia became irreversible when exposed to a mixture of just four non-excitatory amino acids(L-alanine,glycine,L-glutamine,and L-serine)at their plasma concentrations.These amino acids induce swelling in the somas of neurons and astrocytes during hypoxia,along with permanent dendritic damage mediated by N-methyl-D-aspartate receptors.Blocking N-methyl-D-aspartate receptors prevented neuronal damage in the presence of these amino acids during hypoxia.It is likely that astroglial swelling caused by the accumulation of these amino acids via the alanine-serine-cysteine transporter 2 exchanger and system N transporters activates volume-regulated anion channels,leading to the release of excitotoxins and subsequent neuronal damage through N-methyl-D-aspartate receptor activation.Thus,previously unrecognized mechanisms involving non-excitatory amino acids may contribute to the progression and expansion of brain injury in neurological emergencies such as stroke and traumatic brain injury.Understanding these pathways co uld highlight new therapeutic targets to mitigate brain injury.

Low crude protein formulation with supplemental amino acids for its impacts on intestinal health and growth performance of growing-finishing pigs

Background Low crude protein(CP)formulations with supplemental amino acids(AA)are used to enhance intestinal health,reduce costs,minimize environmental impact,and maintain growth performance of pigs.However,extensive reduction of dietary CP can compromise growth performance due to limited synthesis of non-essential AA and limited availability of bioactive compounds from protein supplements even when AA requirements are met.Moreover,implementing a low CP formulation can increase the net energy(NE)content in feeds causing excessive fat deposition.Additional supplementation of functional AA,coupled with low CP formulation could further enhance intestinal health and glucose metabolism,improving nitrogen utilization,and growth performance.Three experiments were conducted to evaluate the effects of low CP formulations with supplemental AA on the intestinal health and growth performance of growing-finishing pigs.Methods In Exp.1,90 pigs(19.7±1.1 kg,45 barrows and 45 gilts)were assigned to 3 treatments:CON(18.0%CP,supplementing Lys,Met,and Thr),LCP(16.0%CP,supplementing Lys,Met,Thr,Trp,and Val),and LCPT(16.1%CP,LCP+0.05%SID Trp).In Exp.2,72 pigs(34.2±4.2 kg BW)were assigned to 3 treatments:CON(17.7%CP,meeting the requirements of Lys,Met,Thr,and Trp);LCP(15.0%CP,meeting Lys,Thr,Trp,Met,Val,Ile,and Phe);and VLCP(12.8%CP,meeting Lys,Thr,Trp,Met,Val,Ile,Phe,His,and Leu).In Exp.3,72 pigs(54.1±5.9 kg BW)were assigned to 3 treatments and fed experimental diets for 3 phases(grower 2,finishing 1,and finishing 2).Treatments were CON(18.0%,13.8%,12.7%CP for 3 phases;meeting Lys,Met,Thr,and Trp);LCP(13.5%,11.4%,10.4%CP for 3 phases;meeting Lys,Thr,Trp,Met,Val,Ile,and Phe);and LCPG(14.1%,12.8%,11.1%CP for 3 phases;LCP+Glu to match SID Glu with CON).All diets had 2.6 Mcal/kg NE.Results In Exp.1,overall,the growth performance did not differ among treatments.The LCPT increased(P<0.05)Claudin-1 expression in the duodenum and jejunum.The LCP and LCPT increased(P<0.05)CAT-1,4F2hc,and B0AT expressions in the jejunum.In Exp.2,overall,the VLCP reduced(P<0.05)G:F and BUN.The LCP and VLCP increased(P<0.05)the backfat thickness(BFT).In Exp.3,overall,growth performance and BFT did not differ among treatments.The LCPG reduced(P<0.05)BUN,whereas increased the insulin in plasma.The LCP and LCPG reduced(P<0.05)the abundance of Streptococcaceae,whereas the LCP reduced(P<0.05)Erysipelotrichaceae,and the alpha diversity.Conclusions When implementing low CP formulation,CP can be reduced by supplementation of Lys,Thr,Met,Trp,Val,and Ile without affecting the growth performance of growing-finishing pigs when NE is adjusted to avoid increased fat deposition.Supplementation of Trp above the requirement or supplementation of Glu in low CP formulation seems to benefit intestinal health as well as improved nitrogen utilization and glucose metabolism.

Six Amino Acids among Natural Moisturizing Factors Responsible for Skin Hydration: Improvement and Anti-Aging of Skin by Galactomyces Ferment Filtrate-PiteraTM Containing Skin Moisturizer

Background: Natural moisturizing factors (NMFs) are filaggrin-derived components in the cornified layer that are critical for maintaining healthy skin moisturization and barrier function. However, studies have reported conflicting findings on the relationship between NMF levels and aging, while few studies have investigated this relationship clinically. To fill this research gap, we determined the levels of major NMF components such as free amino acids, pyrrolidone carboxylic acid, and urocanic acids, and individually verified their relationships with skin hydration, barrier function, age, and skin aging. Purpose: The objective of this study was to clinically investigate the relationship between NMF components levels and skin aging in facial skin. The main NMF components were obtained from facial skin and quantified. We then selected NMF components showing strong relationships to skin hydration, and analyzed the relationships of the levels of these selected NMF components with signs of skin aging, namely, texture, pores, wrinkles, and dullness (L-value). We also examined the efficacy of treatment with a skin care formula (SK-II Facial Treatment Essence, called SK-II FTE hereafter) including Galactomyces ferment filtrate (GFF, PiteraTM) on the selected NMF component levels associated with skin hydration and barrier function, and the signs of skin aging of texture, pores, wrinkles, and dullness (L-value). Method: We conducted two clinical trials in this research. In Study 1, we measured 23 NMF components using tape-stripped cornified layer to quantify them via an HPLC method in 196 Asian females aged 20 to 59 (mean S.D., 38.6 9.4). Facial visual aging parameters [texture, pores, wrinkles, and dullness (L-value)], as well as elasticity (R7), skin hydration, and TEWL, were quantified using facial skin imaging and skin physical property measurement devices. Study 2 was performed to evaluate whether the facial application of SK-II FTE affects the NMF levels and skin aging parameters in 63 Asian female volunteers aged 20 to 55 (38.4 9.03). During the course of Study 2, 0.6 mL of SK-II FTE was applied to the face twice daily in the morning and afternoon. Skin measurements were performed at the start of the day (baseline) and at week 8. Results: In Study 1, we examined the stratum corneum levels of 23 NMF components comparing to the skin hydration status in 196 female subjects. The subjects were divided into two groups using the median of each measured NMF component. Skin hydration values were compared between the two groups defined for each NMF component. The results showed that subjects with higher levels of six amino acids, alanine, arginine, asparagine, glutamine, glycine, and histidine, exhibited significantly higher skin hydration than those with lower amino acid levels. No significant differences in skin hydration values were found for the other 17 NMF components. We then analyzed whether the sum of these six amino acid NMF components (called 6-AA-NMFs, hereafter) is affected by aging. The 6-AA-NMF level peaked in the subjects aged 25-29, and then gradually and significantly decreased with age. Interestingly, the 6-AA-NMF level was significantly correlated with the skin hydration value, but not with TEWL. In addition, the 6-AA-NMF level demonstrated significant correlations with the signs of skin aging of texture, pores, wrinkles, and dullness (L-value). Then, in Study 2, we examined whether the daily application of SK-II FTE affects the 6-AA-NMF level and visual aging parameters in 63 females. SK-II FTE demonstrated significant increases of the levels of 6-AA-NMFs and each of its components associated with hydration and barrier function, and improvements of skin texture, pores, wrinkles, and dullness (L-value) during the 8 weeks of treatment of facial skin. Conclusion: These clinical studies with large numbers of subjects across a wide age range revealed that six amino acids as NMF components were highly correlated with facial skin hydration in the stratum corneum. The levels of these six NMF components were also found to decrease at ages after the 30 s and were significantly correlated with major signs of skin aging. Notably, these six NMF components (6-AA-NMFs) were increased by SK-II FTE treatment associated with improvements of skin hydration and signs of skin aging, namely, texture, pores, wrinkles, and dullness (L-value). These studies were limited by the lack of investigation of why some NMF components were not associated with skin hydration. More clinical trials examining various NMF components and their relationship with aging are anticipated.