Background Diets with high inclusion of corn co-products such as corn fermented protein(CFP)may contain excess Leu,which has a negative impact on feed intake and growth performance of pigs due to increased catabolism ...Background Diets with high inclusion of corn co-products such as corn fermented protein(CFP)may contain excess Leu,which has a negative impact on feed intake and growth performance of pigs due to increased catabolism of Val and Ile and reduced availability of Trp in the brain for serotonin synthesis.However,we hypothesized that the negative effect of using CFP in diets for weanling pigs may be overcome if diets are fortified with crystalline sources of Val,Trp,and(or)Ile.Methods Three hundred and twenty weanling pigs were randomly allotted to one of 10 dietary treatments in a com-pletely randomized design,with 4 pigs per pen and 8 replicate pens per treatment.A corn-soybean meal diet and 2 basal diets based on corn and 10%CFP or corn and 20%CFP were formulated.Seven additional diets were formu-lated by fortifying the basal diet with 20%CFP with Ile,Trp,Val,Ile and Val,Ile and Trp,Trp and Val,or Ile,Trp and Val.A two-phase feeding program was used,with d 1 to 14 being phase 1 and d 15 to 28 being phase 2.Fecal scores were recorded every other day.Blood samples were collected on d 14 and 28 from one pig per pen.On d 14,fecal samples were collected from one pig per pen in 3 of the 10 treatments to determine volatile fatty acids,ammonium concen-tration,and microbial protein.These pigs were also euthanized and ileal tissue was collected.Results There were no effects of dietary treatments on any of the parameters evaluated in phase 1.Inclusion of 10%or 20%CFP in diets reduced(P<0.05)final body weight on d 28,and average daily gain(ADG)and average daily feed intake(ADFI)in phase 2 and for the entire experimental period.However,pigs fed the CFP diet supplemented with Val,Ile,and Trp had final body weight,ADFI,ADG and gain to feed ratio in phase 2 and for the entire experiment that was not different from pigs fed the control diet.Fecal scores in phase 2 were reduced(P<0.05)if CFP was used.Conclusions Corn fermented protein may be included by up to 20%in diets for weanling pigs without affecting growth performance,gut health,or hindgut fermentation,if diets are fortified with extra Val,Trp,and Ile.Inclusion of CFP also improved fecal consistency of pigs.展开更多
Background In this study,the effects of L-leucine(Leu)on rumen fermentation parameters,rumen epithelium development,amino acid composition,rumen bacterial communities and rumen metabolites in beef cattle were investig...Background In this study,the effects of L-leucine(Leu)on rumen fermentation parameters,rumen epithelium development,amino acid composition,rumen bacterial communities and rumen metabolites in beef cattle were investigated.Twenty-four fattening Angus females of similar initial weight(575.5±22.1 kg)were randomly assigned to 2 treatments with 4 replicate pens(3 cattle per pen).They were fed either a basal diet or a basal diet supplemented with 6.0 g L-Leu/100 kg BW/d for 120 d.Results(1)Leu increased the ruminal concentrations of total volatile fatty acid(VFA)(P=0.017),propionate(P=0.023),isovalerate(P=0.001),and branched-chain volatile fatty acid(BCVFA)(P=0.01)at 4 h post-feeding.It also tended to increase acetate(P=0.083)and decrease the ammonia-N(NH_3-N)concentration(P=0.055),but it did not affect ruminal pH(P>0.1).Leu also increased microbial crude protein(MCP)(P=0.026)at 4 h post-feeding,but decreased MCP at 8 h post-feeding(P=0.010).(2)Supplementation with L-Leu increased the ruminal concentrations of phenylalanine(P=0.011),lysine(P=0.034),and tyrosine(P=0.033),while decreasing the cystine concentration(P=0.010).(3)Leu increased the thickness of the stratum spinosum and basal(P<0.05),while decreasing the thickness of the stratum granulosum(P<0.05).(4)Leu upregulated the relative mRNA abundance of genes involved in tight junction proteins(P<0.05)and VFA absorption and metabolism(P<0.01)in the rumen epithelium.This upregulation was positively correlated with the concentrations ruminal isovalerate and BCVFA(P<0.01).(5)L-Leu did not affect the diversity and richness of ruminal microbes(P>0.05),but differential bacterial biomarkers(LEfSe,LDA>2)were either positively or negatively correlated with ruminal MCP,NH_3-N,and BCVFA concentrations(P<0.001).Additionally,differential bacterial metabolites(OPLS-DA,VIP>1.5)were primarily enriched in the amino acid metabolism pathway and the cofactors and vitamins metabolism pathway(P<0.05).Conclusions Dietary supplementation with L-Leu altered rumen fermentation parameters and patterns,improved rumen epithelial morphology,and enhanced the expression of genes related to VFA absorption and metabolism in the rumen epithelium of beef cattle.展开更多
Discoidin domain receptors(DDRs)are single-pass transmembrane proteins belonging to receptor tyrosine kinases(RTKs)family,which are activated by collagen ligands with unusual slow,sustained kinetics,distinguishing the...Discoidin domain receptors(DDRs)are single-pass transmembrane proteins belonging to receptor tyrosine kinases(RTKs)family,which are activated by collagen ligands with unusual slow,sustained kinetics,distinguishing them from canonical RTKs.While DDRs play critical roles in cell adhesion,differentiation,and cancer progression,their activation mechanisms remain partly understood.Here,we investigated the transmembrane domains(TMDs)of DDR1 and DDR2 to elucidate their interaction dynamics in membrane.Using bacterial adenylate cyclase two-hybrid(BACTH)assays,we demonstrated robust homotypic interactions and even stronger heterotypic associations between DDRTMDs.NMR spectroscopy of DDR1TMD and DDR2TMD reconstituted in lipid bilayer-mimetic bicelles showed obvious chemical shift alterations,further validating the stability of their heterocomplex formation.Systematic mutagenesis identified leucine zipper motifs rather than GXXXA motifs mediated both homo-and hetero-associations of DDR1TMD and DDR2TMD.These findings demonstrated the TMD as a critical mediator of DDRs oligomerization and revealed their interaction patterns within membrane.Our study advances the understanding of DDR signaling regulation and highlights transmembrane domain interactions as potential targets for modulating DDR-related pathologies.展开更多
The aim of the present investigation was to develop a biosensor for the detection of amino acids, Leucine, Isoleucine and Valine based on a quartz crystal nanobalance. leucine (Leu), isoleucine (Ile), and valine (Val)...The aim of the present investigation was to develop a biosensor for the detection of amino acids, Leucine, Isoleucine and Valine based on a quartz crystal nanobalance. leucine (Leu), isoleucine (Ile), and valine (Val) were selectively determined by quartz crystal nanobalance (QCN) sensor in conjunction with net analyte signal (NAS)-based method called HLA/GO. An orthogonal design was applied for the formation of calibration and prediction sets including Leu, Ile and Val compounds. The selection of the optimal time range involved the calculation of the net analyte sig-nal regression plot in any considered time window for each test sample. The searching of a region with maximum linearity of NAS regression plot (minimum error indicator) and minimum of PRESS value was carried out by applying a moving window strategy. On the base of obtained results, the differences on the adsorption profiles in the time range between 1 and 300 s were used to determine mixtures of compounds by HLA/GO method. The results showed that the method was successfully applied for the determina-tion of Leu, Ile and Val.展开更多
Neonatal growth is characterized by a high protein synthesis rate that is largely due to an enhanced sensitivity to the postprandial rise in insulin and amino acids, especially leucine. The mechanism of leucine＇s act...Neonatal growth is characterized by a high protein synthesis rate that is largely due to an enhanced sensitivity to the postprandial rise in insulin and amino acids, especially leucine. The mechanism of leucine＇s action in vivo is not well understood. In this study, we investigated the effect of leucine infusion on protein synthesis in skeletal muscle and liver of neonatal pigs. To evaluate the mode of action of leucine, we used rapamycin, an inhibitor of mammalian target of rapamycin （mTOR） complex-1 （mTORC1）. Overnight-fasted 7-day-old piglets were treated with rapamycin for 1 hour and then infused with leucine （400 μmol·kg^-1·h^-1） for 1 hour. Leucine infusion increased the rate of protein synthesis, and ribosomal protein S6 kinase 1 （S6K1） and eukaryotic initiation factor （elF） 4E-binding protein-1 （4E-BP1） phosphorylation in gastrocnemius and masseter muscles （P 〈 0.05）, but not in the liver. The leucine-induced stimulation of protein synthesis and S6K1 and 4E-BP1 phosphorylation were completely blocked by rapamycin, suggesting that leucine action is by an mTORC1-dependent mechanism. Neither leucine nor rapamycin had any effect on the activation of the upstream mTQRC1 regulators, AMP-activated protein kinase and protein kinase B, in skeletal muscle or liver. The activation of elF2α and elongation factor 2 was not affected by leucine or rapamycin, indicating that these two pathways are not limiting steps of leucine-induced protein synthesis. These results suggest that leucine stimulates muscle protein synthesis in neonatal pigs by inducing the activation of mTORC1 and its downstream pathway leading to mRNA translation.展开更多
Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g±0.1 g) was determined using dose-response method.Six isonitogenous (crude protein 43%...Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g±0.1 g) was determined using dose-response method.Six isonitogenous (crude protein 43%) and isoenergetic (19 kJ g-1) practical diets containing six levels of leucine (Diets 1-6) ranging from 1.23% to 4.80% (dry matter) were made at about 0.7% increment of leucine.Equal amino acid nitrogen was maintained by replacing leucine with glutamic acid.Triplicate groups of 60 individuals were fed to apparent satiation by hand twice daily (05:00 and 17:30).The water temperature was 26-32℃, salinity 26-30 and dissolved oxygen approximately 7 mg L-1 during the experimental period.Final weight (FW) of large yellow croaker initially increased with increasing level of dietary leucine but then decreased at further higher level of leucine.The highest FW was obtained in fish fed diet with 3.30% Leucine (Diet 4).FW of fish fed the diet with 4.80% Leucine (Diet 6) was significantly lower than those fed Diet 4.However, no significant differences were observed between the other dietary treatments.Feed efficiency (FE) and whole body composition were independent of dietary leucine contents (P>0.05).The results indicated that leucine was essential for growth of juvenile large yellow croaker.On the basis of FW, the optimum dietary leucine requirement for juvenile large yellow croaker was estimated to be 2.92% of dry matter (6.79% of dietary protein).展开更多
A 56-day feeding trial was conducted to examine the dietary leucine requirement of juvenile Japanese seabass in sea- water floating net cages （1.5 m × 1.5 m × 2.0 m）. Six isonitrogenous （crude protein 40%...A 56-day feeding trial was conducted to examine the dietary leucine requirement of juvenile Japanese seabass in sea- water floating net cages （1.5 m × 1.5 m × 2.0 m）. Six isonitrogenous （crude protein 40%） and isoenergetic （gross energy 20 kJ g-1） diets were formulated to contain different concentrations of leucine （0.9%, 1.49%, 2.07%, 2.70%, 3.30% and 3.88% of dry matter）. Crys- talline L-amino acids were supplemented to simulate the whole body amino acid pattern of Japanese seabass except for leucine. Three groups （30 fish individuals each, 8.0g±0.20g in initial weight） were fed to apparent satiation at 5：00 and 17：30 every day. During the experimental period, the water temperature ranged from 26 to 32℃ and salinity from 26 to 30, and the dissolved oxygen was maintained at 7mgL-l. The results showed that weight gain （WG）, nitrogen retention （NR）, feed efficiency （FE） and protein efficiency ratio （PER） were significantly increased when dietary leucine was increased from 0.90% to 2.70% of dry matter, and then declined. WG was the highest when fish were fed D4 containing 2.70% of leucine. No significant differences were observed in body composition among dietary treatments （P 〉 0.05）. Considering the change of WG, the optimum dietary leucine requirement of juve- nile Japanese seabass was either 2.39% of dry matter or 5.68% of dietary protein.展开更多
The basic leucine zipper （bZIP） transcription factors form a large gene family that is important in pathogen defense, light and stress signaling, etc. The Completed whole genome sequences of model plants Arabidopsis...The basic leucine zipper （bZIP） transcription factors form a large gene family that is important in pathogen defense, light and stress signaling, etc. The Completed whole genome sequences of model plants Arabidopsis （Arabidopsis thaliana）, rice （Oryza sativa） and poplar （Populus trichocarpa） constitute a valuable resource for genome-wide analysis and genomic comparative analysis, as they are representatives of the two major evolutionary lineages within the angiosperms： the monocotyledons and the dicotyledons. In this study, bioinformatics analysis identified 74, 89 and 88 bZIP genes respectively in Arabidopsis, rice and poplar. Moreover, a comprehensive overview of this gene family is presented, including the gene structure, phylogeny, chromosome distribution, conserved motifs. As a result, the plant bZIPs were organized into 10 subfamilies on basis of phylogenetic relationship. Gene duplication events during the family evolution history were also investigated. And it was further concluded that chromosomal/segmental duplication might have played a key role in gene expansion of bZIP gene family.展开更多
The aim of this study is to investigate the effects of leucine（Leu） and histidine（His） on the expression of both the mammalian target of rapamycin（mTOR） signaling pathway-related proteins and caseins in immortal...The aim of this study is to investigate the effects of leucine（Leu） and histidine（His） on the expression of both the mammalian target of rapamycin（mTOR） signaling pathway-related proteins and caseins in immortalized bovine mammary epithelial cells（CMEC-H）, using a single supplement through Western blotting. The Earle＇s balanced salt solution（EBSS） was set as the control group and other treatment groups, based on the EBSS, were added with different concentrations of Leu or His, respectively. The results showed that, compared with the control group, the expression of caseins and the phosphorylation of mTOR（Ser^2481）, Raptor（Ser^792）, e IF4E（Ser^209）, and e EF2（Thr^56） increased with the Leu concentrations ranging from 0.45 to 10.80 mmol/L（P〈0.01）. The P-4EBP1（Thr^37） at 10.80 mmol/L Leu, and P-RPS6（Ser^235/236） at 5.40 to 10.80 mmol/L Leu all decreased. Similarly, the His supplementation from 0.15 to 9.60 mmol/L increased the expression of αs2-casein, β-casein, κ-casein, P-mTOR（Ser^2481）, P-Raptor（Ser^792）, P-S6K1（Thr^389）, P-4EBP1（Thr^37）, P-e IF4E（Ser^209）, and P-e EF2（Thr^56）（P〈0.01） in CMEC-H, whereas the αs1-casein expression was only reduced at 9.60 mmol/L His, G protein β subunit-like protein（GβL） at 0.15 and 9.60 mmol/L His, and P-RPS6 at 4.80 to 9.60 mmol/L His. Our linear regression model assay suggested that the αs1-casein expression was positively correlated with P-mTOR（P〈0.01）, P-S6K1（P〈0.01）, and P-e EF2（P〈0.01） for the addition of Leu, while the expressions of β-casein（P〈0.01） and κ-casein（P〈0.01） were positively correlated with P-e EF2 for the addition of His. In conclusion, the milk protein synthesis was up-regulated through activation of the mTOR pathway with the addition of Leu and His in CMEC-H.展开更多
Epidemiological evidence points to increased dairy and meat consumption,staples of the Western diet,as major risk factors for the development of type 2 diabetes(T2D).This paper presents a new concept and comprehensive...Epidemiological evidence points to increased dairy and meat consumption,staples of the Western diet,as major risk factors for the development of type 2 diabetes(T2D).This paper presents a new concept and comprehensive review of leucine-mediated cell signaling explaining the pathogenesis of T2D and obesity by leucine-induced over-stimulation of mammalian target of rapamycin complex 1(mTORC1).mTORC1,a pivotal nutrient-sensitive kinase,promotes growth and cell proliferation in response to glucose,energy,growth factors and amino acids.Dairy proteins and meat stimulate insulin/insulin-like growth factor 1 signaling and provide high amounts of leucine,a primary and independent stimulator for mTORC1 activation.The downstream target of mTORC1,the kinase S6K1,induces insulin resistance by phosphorylation of insulin receptor substrate-1,thereby increasing the metabolic burden of β-cells.Moreover,leucine-mediated mTORC1-S6K1-signaling plays an important role in adipogenesis,thus increasing the risk of obesity-mediated insulin resistance. High consumption of leucine-rich proteins explains exaggerated mTORC1-dependent insulin secretion, increased β-cell growth and β-cell proliferation promoting an early onset of replicative β-cell senescence with subsequent β-cell apoptosis.Disturbances of β-cell mass regulation with increased β-cell proliferation and apoptosis as well as insulin resistance are hallmarks of T2D,which are all associated with hyperactivation of mTORC1.In contrast,the anti-diabetic drug metformin antagonizes leucine-mediated mTORC1 signaling.Plant-derived polyphenols and flavonoids are identified as natural inhibitors of mTORC1 and exert anti-diabetic and anti-obesity effects.Furthermore,bariatric surgery in obesity reduces increased plasma levels of leucine and other branched-chain amino acids.Attenuation of leucine-mediated mTORC1 signaling by defining appropriate upper limits of the daily intake of leucine-rich animal and dairy proteins may offer a great chance for the prevention of T2D and obesity,as well as other epidemic diseases of civilization with increased mTORC1 signaling,especially cancer and neurodegenerative diseases,which are frequently associated with T2D.展开更多
Low protein intake causes a decrease in protein deposition in most animal tissues. The purpose of this study was to investigate whether leucine supplementation would increase the synthesis rate of protein and muscle w...Low protein intake causes a decrease in protein deposition in most animal tissues. The purpose of this study was to investigate whether leucine supplementation would increase the synthesis rate of protein and muscle weight in adult rats, which chronically consume only 58.8% of their protein requirements. Thirty-six male Sprague-Dawley rats were assigned to one of three dietary treatments including a 20% casein diet （CON）, a 10% casein ＋ 0.44% alanine diet （R）, and a 10% casein ＋ 0.87% leucine diet （RL）. After a 10 d dietary treatment, plasma amino acid levels were measured after feeding, the gastrocnemius muscles and soleus muscles were harvested and weighed, and the fractional synthesis rate （FSR） and mammalian target of rapamycin （mTOR） signaling proteins in skeletal muscle were measured. Regarding the plasma amino acid level, the RL group had the highest concentration of leucine （P 〈 0.05） and the lowest concentration of isoleucine （P 〈 0.05） among the three groups, and the CON group had a lower concentration of valine （P 〈 0.05） than the R and RL groups. Compared with the R and RE groups, the CON group diet significantly increased （P 〈 0.05） feed intake, protein synthesis rate, and the phosphorylation of eukaryutic initiation factor 4E binding protein 1 （4E-BP1）, and decreased the weight of abdominal adipose. Compared with the R group, the RL group significantly increased in gastrocnemius muscle weight, protein synthesis rate, and phosphorylation of both ribosomal protein $6 kinase 1 （56K1） and 4E-BP1. In conclusion, when protein is chronically restricted in adult rat diets, leucine supplementation moderately improves body weight gain and increases muscle protein synthesis through mTOR activation,展开更多
Bacterial production is one of the key parameters to evaluate bacterial role in ocean carbon fluxes.Estimation of bacterial production requires the leucine-to-carbon conversion factors that change widely across enviro...Bacterial production is one of the key parameters to evaluate bacterial role in ocean carbon fluxes.Estimation of bacterial production requires the leucine-to-carbon conversion factors that change widely across environments.However,empirical leucine-to-carbon conversion factors（e CFs） are seldom determined in situ because of time consuming and little is known on regulating factors for the e CFs.During May 2015 to January 2016,fourteen dilution experiments were conducted,from the Zhujiang（Pearl River） Estuary to the coast of the northern South China Sea,to determine spatiotemporal variability in the e CFs and its potential controlling factors along an environmental gradient.The e CFs showed clear spatial variations with the highest（1.27–1.69（kg C）/（mol Leu）） in low salinity waters（salinity〈15）,intermediate（1.03–1.25（kg C）/（mol Leu）） in moderate salinity（salinity of 15–25）,and the lowest（0.48–0.85（kg C）/（mol Leu）） in high salinity waters（salinity〉25）.Substrate availability was responsible for spatial variability in the e CFs.In the pristine coastal waters,low e CFs was related to substrate limitation and leucine incorporated was respired to maximize the survival rather than bacterial production.Hence,the e CFs measurement was needed for estimating bacterial production accurately in various marine environments.展开更多
Genetic control of leucine content in indica-japonica hybrid rice (Oryza sativa L.) was studied in 35 crosses of F1 and F2 generations, which were derived from crossing 7 male sterile indica rice lines with 5 restorer...Genetic control of leucine content in indica-japonica hybrid rice (Oryza sativa L.) was studied in 35 crosses of F1 and F2 generations, which were derived from crossing 7 male sterile indica rice lines with 5 restorer japonica rice lines along with their parents. Two genetic models and their corresponding statistical methods for quantitative traits of triploid seeds in cereal crops were used for the analysis. The first was the unconditional genetic model, which refers to the analysis of cumulative measurements (from flowering to a specific time) along the developmental stages, while the second was the conditional genetic model, which relates to analysis from one developmental stage to another stage (t - 1→t). The results showed that leucine content of indica-japonica hybrid rice was controlled by the expression of triploid endosperm effect (endosperm additive effect and endosperm dominant effect), cytoplasm effect, diploid maternal plant effect (maternal additive effect and maternal dominant effect) and their environmental interaction effects. Of these effects, endosperm dominant effect and maternal dominant effect were more important at the earlier stages, while endosperm additive effect and maternal additive effect were more important at the later stages of rice grain development under both unconditional and conditional genetic analyses. Due to the high heritabilities, which came from endosperm, maternal and cytoplasm effects for leucine content at different developmental stages, selection for leucine content of indica-japonica hybrid rice would be more efficient at early generations in breeding programs.展开更多
Objective: Reactive oxygen species (ROS) are involved in the endothelial-mediated disorders within atherosclerosis. Considering that an oxidant/antioxidant imbalance might be a key factor in the damaging ROS-mediated ...Objective: Reactive oxygen species (ROS) are involved in the endothelial-mediated disorders within atherosclerosis. Considering that an oxidant/antioxidant imbalance might be a key factor in the damaging ROS-mediated effects, the present study intends to determine the influence of a high-fat diet, associated with essential amino acids—valine and leucine, upon the experimental animals, through evaluation of plasmatic level of some antioxidant enzymes. Material and Methods: The study was conducted on 32 male Wistar rats, which were fed with cholesterol, valine and leucine, for 60 days. The animals were divided into four groups, according to the received diet: the first group—standard diet;the second group—cholesterol (C);the third group—cholesterol and valine (C + V);the fourth group—cholesterol and leucine (C + L). Evaluations of the oxidative status, through plasma levels of the antioxidant enzymes: superoxide dismutase (SOD) and glutathione peroxidise (GPx), were made for the four mentioned groups of animals, at the beginning of the study (R0), after one (R1) and two months (R2). Results: The average values of SOD and GPx in group of animals fed exclusively with cholesterol (C) were significantly higher compared to the third group where cholesterol was supplemented with valine (C + V) or fourth group fed with cholesterol and leucine (C + L) (p < 0.001), after one month as well at the end of the experiment (two months). There were no significant differences in the levels of SOD and GPx between group III and group IV (p < 0.05) at the end of the experiment. Conclusion: Our results showed that valine and leucine decreased the serum levels of SOD and GPx and therefore they were useful antioxidants, which could improve the endothelial dysfunctions associated with atherosclerosis. Moreover, analysis of the oxidative status in the context of atherosclerotic mediated endothelial damage suggests that deviation from normal to alter endothelial status may be conditioned by an oxidants/antioxidants imbalance.展开更多
Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) co...Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) constitute a family of transcription factors that are characterized by the presence of a DNA-binding domain closely linked with leucine zipper motif functioning in dimer formation. This type of association is unique to plants and considered as an excellent candidate to activate developmental responses to altering environmental conditions. Cotton is the most important fiber plant with a lot of local and commercial uses in the world. HD-Zip proteins not only have key roles in different stages of vascular and inter-fascicular fiber differentiation of cotton but also are suggested to have an important role against abiotic stress that is one of the key factors limiting cotton productivity. Plants have developed various strategies to manage stress conditions through a combination of metabolic, physiological and morphological adaptations. These adaptive changes rely largely on alterations in gene expression. Therefore, transcriptional regulators play a crucial role in stress tolerance. Being a transcription factor HD-Zip might be a useful target for genetic engineering to generate multiple stress tolerance in susceptible plants. In the following chapter, we discussed how the HD-Zip proteins would play a useful role for cotton development both in fiber production and stress adaptation.展开更多
An inadequate amino acid (AA) supply in animals under protein-restricted conditions can slow skeletal muscle growth.Protein translation can be activated by short-term leucine (Leu) stimulation;however,whether muscle m...An inadequate amino acid (AA) supply in animals under protein-restricted conditions can slow skeletal muscle growth.Protein translation can be activated by short-term leucine (Leu) stimulation;however,whether muscle mass increases under long-term Leu supplementation and how the gut and muscle respond to Leu supplementation are largely unknown.In this study,we investigated if muscle mass increases with long-term Leu supplementation under protein-restricted conditions.We identified changes in the link between the gut and muscles under different amino acid supply conditions,using goats as the study object.A total of 27 Xiangdong black male goats with average initial body weight (BW)of 10.88±1.22 kg were randomly divided into three dietary treatments:a normal protein diet (NP,14.24%crude protein[CP]);a low protein diet (LP,8.27%CP with supplemental 1.66%rumen-protected lysine[RPLys]and 0.09%rumen-protected methionine[RPMet]);and LP diet with rumen-protected Leu (RPLeu)(LP+RPLeu,8.75%CP with supplemental 1.66%RPLys,0.09%RPMet and 1.46%RPLeu).The animal trial lasted for 110 d,consisting of 20 d of adaptation and a 90 d of experimental period.The results showed that long-term protein restriction increased gut tryptophan hydroxylase 1 (TPH1) activity (P<0.001),tryptophan (Trp) catabolism (P<0.001),and 5-hydroxytryptamine (5-HT) synthesis (P<0.001),which all subsequently reduced goat appetite.Long-term Leu supplementation inhibited 5-HT synthesis(P<0.001),decreased Trp catabolism in the gut,and increased appetite in goats.Long-term protein restriction enhanced jejunal and ileal branched-chain amino acid transferase (BCAT)(P<0.001) and branched-chain a-Keto acid dehydrogenase (BCKD)(P=0.048) activities,which increased branchedchain amino acid (BCAA) catabolism.Immunofluorescence results showed that protein restriction decreased the intestinal mucosal expression of solute carrier family 1 member 5 (SLC1A5)(P=0.032) and solute carrier family 7 member 5 (SLC7A5)(P<0.001),reduced BCAA transport from the mucosa to the blood,lowered BCAA levels in the blood (P<0.001).Western blot results showed that protein restriction inhibited mammalian target of rapamycin (mTOR) pathway activation in goat muscles.Leu supplementation increased BCAA translocation from the intestine to the blood and promoted activation of the muscle m TOR pathway and protein synthesis.In conclusion,our results suggest that Leu supplementation in low-protein diets improves appetite and alleviates the inhibition of muscle protein synthesis in goats.展开更多
Leucine-rich repeat containing 15(LRRC15)has emerged as an attractive biomarker and target for cancer therapy.Transforming growth factor-β(TGFβ)induces the expression of this plasma membrane protein specifically in ...Leucine-rich repeat containing 15(LRRC15)has emerged as an attractive biomarker and target for cancer therapy.Transforming growth factor-β(TGFβ)induces the expression of this plasma membrane protein specifically in aggressive and treatment resistant tumor cells derived from mesenchymal stem cells,with minimal expression observed in non-neoplastic tissues.We have developed a humanized monoclonal antibody,DUNP19,that specifically binds with high affinity to a phylogenetically conserved LRRC15 epitope and is rapidly internalized upon LRRC15 binding.In multiple subcutaneous and orthotopic tumor xenograft mouse models,Lutetium-177 labeled DUNP19([^(177)Lu]Lu-DUNP19)enabled non-invasive imaging and molecularly precise radiotherapy to LRRC15-expressing cancer cells and murine cancer-associated fibroblasts,effectively halting tumor progression and prolonging survival with minimal toxicity.Transcriptomic analyses of[^(177)Lu]Lu-DUNP19-treated tumors reveal a loss of pro-tumorigenic mechanisms,including a previously reported TGF β-induced LRRC15+signature associated with immunotherapy resistance.In a syngeneic tumor model,administration of[^(177)Lu]Lu-DUNP19 significantly potentiated checkpoint-blockade therapy,yielding durable complete responses.Together,these results demonstrate that radio-theranostic targeting of LRRC15 with DUNP19 is a compelling precision medicine platform for image-guided diagnosis,eradication,and reprogramming of LRRC15+tumor tissue that drives immunoresistance and disease aggressiveness in a wide range of currently untreatable malignancies.展开更多
In this study, ion mobility separation coupled with tandem mass spectrometry （IM-tandem MS） was utilized to investigate the ionization behaviors of two amino acids including leucine and isoleucine. Under the electro...In this study, ion mobility separation coupled with tandem mass spectrometry （IM-tandem MS） was utilized to investigate the ionization behaviors of two amino acids including leucine and isoleucine. Under the electrospray ionization （ESI） mode, two protonation sites in each molecular sturcture caused two forms of protomer. One arose from the amino being protonated （amino-protomer） and the other from the carboxyl being protonated （car- boxyl-protomer）. In the two-dimensional （drift time, m/z） spectrum, the protomers had the same mass, but the dis- tinguishable drift times and fragmentation patterns. For the characterization purpose, the theoretical collision cross section （CCS） values of the protomers were calculated and proven to be consistent with the experimental. Moreover, the quantified relationship between the amino acids and their protomers was evaluated. It showed that the abun- dance of the carboxyl-protomer was proportional to the concentration of the amino acid, whereas that of the amino- protomer did not have the same trend. Under the atmospheric pressure chemical ionization （APCI） mode, only the carboxyl-protomer was observed. In addition, the amino-protomer and the cluster ions observed under ESI were absent completely. The results demonstrate that the ionization mode impacts heavily on the ionization behaviors of leucine and isoleucine not only on the form of therir protomers but also On the quantified relationship.展开更多
Soy whey is a liquid by-product generated from tofu production and it is often discarded as wastewater after it is generated.Previous studies have shown that out of the three branched-chain amino acids,isoleucine was ...Soy whey is a liquid by-product generated from tofu production and it is often discarded as wastewater after it is generated.Previous studies have shown that out of the three branched-chain amino acids,isoleucine was found to have an inhibitory effect on Torulaspora delbrueckii Biodiva during soy whey fermentation.In this study,a comparison between isoleucine and leucine was carried out to ascertain how these two structural isomers could affect the growth and metabolism of T.delbrueckii Biodiva at various concentrations(40,80 and 160 mg nitrogen/L).In general,amino acid supplementation sped up the fermentation process relative to unsupplemented samples.Even though glycerol and ethanol production were enhanced in all isoleucine supplemented samples in comparison to unsupplemented samples,isoleucine supplementation resulted in slower yeast growth at day 1 and slower sucrose hydrolysis regardless of the concentration supplemented.As for leucine supplementation,there was no impact on yeast growth,glycerol and ethanol production but sucrose hydrolysis was enhanced in comparison to the unsupplemented samples.Increasing isoleucine and leucine supplementation resulted in corresponding increase in active amyl alcohol and isoamyl alcohol production respectively,but isoleucine had a stimulating effect on increasing the production of other higher alcohols while leucine had an inhibitory effect on the production of other higher alcohols.Therefore,even though isoleucine and leucine are structural isomers with the positional difference of only one methyl group,the impact on the yeast metabolism and growth is visibly different.展开更多
The Arabidopsis （Arabidopsis thaliana L.） genome encodes for four distinct classes of homeodomain leucinezipper （HD-ZIP） transcription factors （HD-ZIPI to HD-ZIPIV）, which are all organized in multi-gene familie...The Arabidopsis （Arabidopsis thaliana L.） genome encodes for four distinct classes of homeodomain leucinezipper （HD-ZIP） transcription factors （HD-ZIPI to HD-ZIPIV）, which are all organized in multi-gene families. HD-ZIP transcription factors act as sequence-specific DNA-binding proteins that are able to control the expression level of target genes. While HD-ZIPI and HD-ZIPII proteins are mainly associated with environmental responses, HD-ZIPIII and HD- ZIPIV are primarily known to act as patterning factors. Recent studies have challenged this view. It appears that several of the different HD-ZlP families interact genetically to align both morphogenesis and environmental responses, most likely by modulating phytohormone-signaling networks.展开更多
基金Financial support for this research from Minnesota Corn Growers Association(Burnsville,MN,USA)Green Plains Energy(Omaha,NE,USA)is greatly appreciated.
文摘Background Diets with high inclusion of corn co-products such as corn fermented protein(CFP)may contain excess Leu,which has a negative impact on feed intake and growth performance of pigs due to increased catabolism of Val and Ile and reduced availability of Trp in the brain for serotonin synthesis.However,we hypothesized that the negative effect of using CFP in diets for weanling pigs may be overcome if diets are fortified with crystalline sources of Val,Trp,and(or)Ile.Methods Three hundred and twenty weanling pigs were randomly allotted to one of 10 dietary treatments in a com-pletely randomized design,with 4 pigs per pen and 8 replicate pens per treatment.A corn-soybean meal diet and 2 basal diets based on corn and 10%CFP or corn and 20%CFP were formulated.Seven additional diets were formu-lated by fortifying the basal diet with 20%CFP with Ile,Trp,Val,Ile and Val,Ile and Trp,Trp and Val,or Ile,Trp and Val.A two-phase feeding program was used,with d 1 to 14 being phase 1 and d 15 to 28 being phase 2.Fecal scores were recorded every other day.Blood samples were collected on d 14 and 28 from one pig per pen.On d 14,fecal samples were collected from one pig per pen in 3 of the 10 treatments to determine volatile fatty acids,ammonium concen-tration,and microbial protein.These pigs were also euthanized and ileal tissue was collected.Results There were no effects of dietary treatments on any of the parameters evaluated in phase 1.Inclusion of 10%or 20%CFP in diets reduced(P<0.05)final body weight on d 28,and average daily gain(ADG)and average daily feed intake(ADFI)in phase 2 and for the entire experimental period.However,pigs fed the CFP diet supplemented with Val,Ile,and Trp had final body weight,ADFI,ADG and gain to feed ratio in phase 2 and for the entire experiment that was not different from pigs fed the control diet.Fecal scores in phase 2 were reduced(P<0.05)if CFP was used.Conclusions Corn fermented protein may be included by up to 20%in diets for weanling pigs without affecting growth performance,gut health,or hindgut fermentation,if diets are fortified with extra Val,Trp,and Ile.Inclusion of CFP also improved fecal consistency of pigs.
基金supported by the National Key R&D Program(Grant No.2022YFD1301101-1)National Natural Science Foundation of China(Grant No.32172758)Key Research and Development Program of Ningxia(Grant No.2024BBF01008)。
文摘Background In this study,the effects of L-leucine(Leu)on rumen fermentation parameters,rumen epithelium development,amino acid composition,rumen bacterial communities and rumen metabolites in beef cattle were investigated.Twenty-four fattening Angus females of similar initial weight(575.5±22.1 kg)were randomly assigned to 2 treatments with 4 replicate pens(3 cattle per pen).They were fed either a basal diet or a basal diet supplemented with 6.0 g L-Leu/100 kg BW/d for 120 d.Results(1)Leu increased the ruminal concentrations of total volatile fatty acid(VFA)(P=0.017),propionate(P=0.023),isovalerate(P=0.001),and branched-chain volatile fatty acid(BCVFA)(P=0.01)at 4 h post-feeding.It also tended to increase acetate(P=0.083)and decrease the ammonia-N(NH_3-N)concentration(P=0.055),but it did not affect ruminal pH(P>0.1).Leu also increased microbial crude protein(MCP)(P=0.026)at 4 h post-feeding,but decreased MCP at 8 h post-feeding(P=0.010).(2)Supplementation with L-Leu increased the ruminal concentrations of phenylalanine(P=0.011),lysine(P=0.034),and tyrosine(P=0.033),while decreasing the cystine concentration(P=0.010).(3)Leu increased the thickness of the stratum spinosum and basal(P<0.05),while decreasing the thickness of the stratum granulosum(P<0.05).(4)Leu upregulated the relative mRNA abundance of genes involved in tight junction proteins(P<0.05)and VFA absorption and metabolism(P<0.01)in the rumen epithelium.This upregulation was positively correlated with the concentrations ruminal isovalerate and BCVFA(P<0.01).(5)L-Leu did not affect the diversity and richness of ruminal microbes(P>0.05),but differential bacterial biomarkers(LEfSe,LDA>2)were either positively or negatively correlated with ruminal MCP,NH_3-N,and BCVFA concentrations(P<0.001).Additionally,differential bacterial metabolites(OPLS-DA,VIP>1.5)were primarily enriched in the amino acid metabolism pathway and the cofactors and vitamins metabolism pathway(P<0.05).Conclusions Dietary supplementation with L-Leu altered rumen fermentation parameters and patterns,improved rumen epithelial morphology,and enhanced the expression of genes related to VFA absorption and metabolism in the rumen epithelium of beef cattle.
基金supported by the National Natural Science Foundation of China(32471354 to T.C.and 82260400 to J.L)Natural Science Foundation of Hainan Province(No.822RC703 to J.L)。
文摘Discoidin domain receptors(DDRs)are single-pass transmembrane proteins belonging to receptor tyrosine kinases(RTKs)family,which are activated by collagen ligands with unusual slow,sustained kinetics,distinguishing them from canonical RTKs.While DDRs play critical roles in cell adhesion,differentiation,and cancer progression,their activation mechanisms remain partly understood.Here,we investigated the transmembrane domains(TMDs)of DDR1 and DDR2 to elucidate their interaction dynamics in membrane.Using bacterial adenylate cyclase two-hybrid(BACTH)assays,we demonstrated robust homotypic interactions and even stronger heterotypic associations between DDRTMDs.NMR spectroscopy of DDR1TMD and DDR2TMD reconstituted in lipid bilayer-mimetic bicelles showed obvious chemical shift alterations,further validating the stability of their heterocomplex formation.Systematic mutagenesis identified leucine zipper motifs rather than GXXXA motifs mediated both homo-and hetero-associations of DDR1TMD and DDR2TMD.These findings demonstrated the TMD as a critical mediator of DDRs oligomerization and revealed their interaction patterns within membrane.Our study advances the understanding of DDR signaling regulation and highlights transmembrane domain interactions as potential targets for modulating DDR-related pathologies.
文摘The aim of the present investigation was to develop a biosensor for the detection of amino acids, Leucine, Isoleucine and Valine based on a quartz crystal nanobalance. leucine (Leu), isoleucine (Ile), and valine (Val) were selectively determined by quartz crystal nanobalance (QCN) sensor in conjunction with net analyte signal (NAS)-based method called HLA/GO. An orthogonal design was applied for the formation of calibration and prediction sets including Leu, Ile and Val compounds. The selection of the optimal time range involved the calculation of the net analyte sig-nal regression plot in any considered time window for each test sample. The searching of a region with maximum linearity of NAS regression plot (minimum error indicator) and minimum of PRESS value was carried out by applying a moving window strategy. On the base of obtained results, the differences on the adsorption profiles in the time range between 1 and 300 s were used to determine mixtures of compounds by HLA/GO method. The results showed that the method was successfully applied for the determina-tion of Leu, Ile and Val.
文摘Neonatal growth is characterized by a high protein synthesis rate that is largely due to an enhanced sensitivity to the postprandial rise in insulin and amino acids, especially leucine. The mechanism of leucine＇s action in vivo is not well understood. In this study, we investigated the effect of leucine infusion on protein synthesis in skeletal muscle and liver of neonatal pigs. To evaluate the mode of action of leucine, we used rapamycin, an inhibitor of mammalian target of rapamycin （mTOR） complex-1 （mTORC1）. Overnight-fasted 7-day-old piglets were treated with rapamycin for 1 hour and then infused with leucine （400 μmol·kg^-1·h^-1） for 1 hour. Leucine infusion increased the rate of protein synthesis, and ribosomal protein S6 kinase 1 （S6K1） and eukaryotic initiation factor （elF） 4E-binding protein-1 （4E-BP1） phosphorylation in gastrocnemius and masseter muscles （P 〈 0.05）, but not in the liver. The leucine-induced stimulation of protein synthesis and S6K1 and 4E-BP1 phosphorylation were completely blocked by rapamycin, suggesting that leucine action is by an mTORC1-dependent mechanism. Neither leucine nor rapamycin had any effect on the activation of the upstream mTQRC1 regulators, AMP-activated protein kinase and protein kinase B, in skeletal muscle or liver. The activation of elF2α and elongation factor 2 was not affected by leucine or rapamycin, indicating that these two pathways are not limiting steps of leucine-induced protein synthesis. These results suggest that leucine stimulates muscle protein synthesis in neonatal pigs by inducing the activation of mTORC1 and its downstream pathway leading to mRNA translation.
基金supported by the National Key Technologies Research and Development Program for the 10th Five-year Plan of China (Grant No.2004BA526B-06)Program for New Century Excellent Talents in University (NCET-07-0776)
文摘Dietary leucine requirement for juvenile large yellow croaker, Pseudosciaena crocea Richardson 1846 (initial body weight 6.0 g±0.1 g) was determined using dose-response method.Six isonitogenous (crude protein 43%) and isoenergetic (19 kJ g-1) practical diets containing six levels of leucine (Diets 1-6) ranging from 1.23% to 4.80% (dry matter) were made at about 0.7% increment of leucine.Equal amino acid nitrogen was maintained by replacing leucine with glutamic acid.Triplicate groups of 60 individuals were fed to apparent satiation by hand twice daily (05:00 and 17:30).The water temperature was 26-32℃, salinity 26-30 and dissolved oxygen approximately 7 mg L-1 during the experimental period.Final weight (FW) of large yellow croaker initially increased with increasing level of dietary leucine but then decreased at further higher level of leucine.The highest FW was obtained in fish fed diet with 3.30% Leucine (Diet 4).FW of fish fed the diet with 4.80% Leucine (Diet 6) was significantly lower than those fed Diet 4.However, no significant differences were observed between the other dietary treatments.Feed efficiency (FE) and whole body composition were independent of dietary leucine contents (P>0.05).The results indicated that leucine was essential for growth of juvenile large yellow croaker.On the basis of FW, the optimum dietary leucine requirement for juvenile large yellow croaker was estimated to be 2.92% of dry matter (6.79% of dietary protein).
基金supported by the National Key Technologies R&D Program for the 15th Five-year Plan of China (Grant no. 2004BA526B-06)Program for New Century Excellent Talents in University (NCET-07-0776)
文摘A 56-day feeding trial was conducted to examine the dietary leucine requirement of juvenile Japanese seabass in sea- water floating net cages （1.5 m × 1.5 m × 2.0 m）. Six isonitrogenous （crude protein 40%） and isoenergetic （gross energy 20 kJ g-1） diets were formulated to contain different concentrations of leucine （0.9%, 1.49%, 2.07%, 2.70%, 3.30% and 3.88% of dry matter）. Crys- talline L-amino acids were supplemented to simulate the whole body amino acid pattern of Japanese seabass except for leucine. Three groups （30 fish individuals each, 8.0g±0.20g in initial weight） were fed to apparent satiation at 5：00 and 17：30 every day. During the experimental period, the water temperature ranged from 26 to 32℃ and salinity from 26 to 30, and the dissolved oxygen was maintained at 7mgL-l. The results showed that weight gain （WG）, nitrogen retention （NR）, feed efficiency （FE） and protein efficiency ratio （PER） were significantly increased when dietary leucine was increased from 0.90% to 2.70% of dry matter, and then declined. WG was the highest when fish were fed D4 containing 2.70% of leucine. No significant differences were observed in body composition among dietary treatments （P 〉 0.05）. Considering the change of WG, the optimum dietary leucine requirement of juve- nile Japanese seabass was either 2.39% of dry matter or 5.68% of dietary protein.
文摘The basic leucine zipper （bZIP） transcription factors form a large gene family that is important in pathogen defense, light and stress signaling, etc. The Completed whole genome sequences of model plants Arabidopsis （Arabidopsis thaliana）, rice （Oryza sativa） and poplar （Populus trichocarpa） constitute a valuable resource for genome-wide analysis and genomic comparative analysis, as they are representatives of the two major evolutionary lineages within the angiosperms： the monocotyledons and the dicotyledons. In this study, bioinformatics analysis identified 74, 89 and 88 bZIP genes respectively in Arabidopsis, rice and poplar. Moreover, a comprehensive overview of this gene family is presented, including the gene structure, phylogeny, chromosome distribution, conserved motifs. As a result, the plant bZIPs were organized into 10 subfamilies on basis of phylogenetic relationship. Gene duplication events during the family evolution history were also investigated. And it was further concluded that chromosomal/segmental duplication might have played a key role in gene expansion of bZIP gene family.
基金Project supported by the National Basic Research Program(973)of China(No.2011CB100805)the Modern Agro-Industry Technology Research System of China(No.nycytx-04-01)the Agricultural Science and Technology Innovation Program(No.ASTIP-IAS12),China
文摘The aim of this study is to investigate the effects of leucine（Leu） and histidine（His） on the expression of both the mammalian target of rapamycin（mTOR） signaling pathway-related proteins and caseins in immortalized bovine mammary epithelial cells（CMEC-H）, using a single supplement through Western blotting. The Earle＇s balanced salt solution（EBSS） was set as the control group and other treatment groups, based on the EBSS, were added with different concentrations of Leu or His, respectively. The results showed that, compared with the control group, the expression of caseins and the phosphorylation of mTOR（Ser^2481）, Raptor（Ser^792）, e IF4E（Ser^209）, and e EF2（Thr^56） increased with the Leu concentrations ranging from 0.45 to 10.80 mmol/L（P〈0.01）. The P-4EBP1（Thr^37） at 10.80 mmol/L Leu, and P-RPS6（Ser^235/236） at 5.40 to 10.80 mmol/L Leu all decreased. Similarly, the His supplementation from 0.15 to 9.60 mmol/L increased the expression of αs2-casein, β-casein, κ-casein, P-mTOR（Ser^2481）, P-Raptor（Ser^792）, P-S6K1（Thr^389）, P-4EBP1（Thr^37）, P-e IF4E（Ser^209）, and P-e EF2（Thr^56）（P〈0.01） in CMEC-H, whereas the αs1-casein expression was only reduced at 9.60 mmol/L His, G protein β subunit-like protein（GβL） at 0.15 and 9.60 mmol/L His, and P-RPS6 at 4.80 to 9.60 mmol/L His. Our linear regression model assay suggested that the αs1-casein expression was positively correlated with P-mTOR（P〈0.01）, P-S6K1（P〈0.01）, and P-e EF2（P〈0.01） for the addition of Leu, while the expressions of β-casein（P〈0.01） and κ-casein（P〈0.01） were positively correlated with P-e EF2 for the addition of His. In conclusion, the milk protein synthesis was up-regulated through activation of the mTOR pathway with the addition of Leu and His in CMEC-H.
基金Supported by Bertelsmann Foundation Gütersloh,Germany
文摘Epidemiological evidence points to increased dairy and meat consumption,staples of the Western diet,as major risk factors for the development of type 2 diabetes(T2D).This paper presents a new concept and comprehensive review of leucine-mediated cell signaling explaining the pathogenesis of T2D and obesity by leucine-induced over-stimulation of mammalian target of rapamycin complex 1(mTORC1).mTORC1,a pivotal nutrient-sensitive kinase,promotes growth and cell proliferation in response to glucose,energy,growth factors and amino acids.Dairy proteins and meat stimulate insulin/insulin-like growth factor 1 signaling and provide high amounts of leucine,a primary and independent stimulator for mTORC1 activation.The downstream target of mTORC1,the kinase S6K1,induces insulin resistance by phosphorylation of insulin receptor substrate-1,thereby increasing the metabolic burden of β-cells.Moreover,leucine-mediated mTORC1-S6K1-signaling plays an important role in adipogenesis,thus increasing the risk of obesity-mediated insulin resistance. High consumption of leucine-rich proteins explains exaggerated mTORC1-dependent insulin secretion, increased β-cell growth and β-cell proliferation promoting an early onset of replicative β-cell senescence with subsequent β-cell apoptosis.Disturbances of β-cell mass regulation with increased β-cell proliferation and apoptosis as well as insulin resistance are hallmarks of T2D,which are all associated with hyperactivation of mTORC1.In contrast,the anti-diabetic drug metformin antagonizes leucine-mediated mTORC1 signaling.Plant-derived polyphenols and flavonoids are identified as natural inhibitors of mTORC1 and exert anti-diabetic and anti-obesity effects.Furthermore,bariatric surgery in obesity reduces increased plasma levels of leucine and other branched-chain amino acids.Attenuation of leucine-mediated mTORC1 signaling by defining appropriate upper limits of the daily intake of leucine-rich animal and dairy proteins may offer a great chance for the prevention of T2D and obesity,as well as other epidemic diseases of civilization with increased mTORC1 signaling,especially cancer and neurodegenerative diseases,which are frequently associated with T2D.
文摘Low protein intake causes a decrease in protein deposition in most animal tissues. The purpose of this study was to investigate whether leucine supplementation would increase the synthesis rate of protein and muscle weight in adult rats, which chronically consume only 58.8% of their protein requirements. Thirty-six male Sprague-Dawley rats were assigned to one of three dietary treatments including a 20% casein diet （CON）, a 10% casein ＋ 0.44% alanine diet （R）, and a 10% casein ＋ 0.87% leucine diet （RL）. After a 10 d dietary treatment, plasma amino acid levels were measured after feeding, the gastrocnemius muscles and soleus muscles were harvested and weighed, and the fractional synthesis rate （FSR） and mammalian target of rapamycin （mTOR） signaling proteins in skeletal muscle were measured. Regarding the plasma amino acid level, the RL group had the highest concentration of leucine （P 〈 0.05） and the lowest concentration of isoleucine （P 〈 0.05） among the three groups, and the CON group had a lower concentration of valine （P 〈 0.05） than the R and RL groups. Compared with the R and RE groups, the CON group diet significantly increased （P 〈 0.05） feed intake, protein synthesis rate, and the phosphorylation of eukaryutic initiation factor 4E binding protein 1 （4E-BP1）, and decreased the weight of abdominal adipose. Compared with the R group, the RL group significantly increased in gastrocnemius muscle weight, protein synthesis rate, and phosphorylation of both ribosomal protein $6 kinase 1 （56K1） and 4E-BP1. In conclusion, when protein is chronically restricted in adult rat diets, leucine supplementation moderately improves body weight gain and increases muscle protein synthesis through mTOR activation,
基金The National Key Research and Development Program under contract No.2016YFA0601203the National Natural Science Foundation of China under contract Nos 41476137 and 41676075+1 种基金the Project of State Key Laboratory of Tropical Oceanography under contract No.LTOZZ1504-1the Hundred Talent Program of Chinese Academy of Sciences under contract No.Y35L041001
文摘Bacterial production is one of the key parameters to evaluate bacterial role in ocean carbon fluxes.Estimation of bacterial production requires the leucine-to-carbon conversion factors that change widely across environments.However,empirical leucine-to-carbon conversion factors（e CFs） are seldom determined in situ because of time consuming and little is known on regulating factors for the e CFs.During May 2015 to January 2016,fourteen dilution experiments were conducted,from the Zhujiang（Pearl River） Estuary to the coast of the northern South China Sea,to determine spatiotemporal variability in the e CFs and its potential controlling factors along an environmental gradient.The e CFs showed clear spatial variations with the highest（1.27–1.69（kg C）/（mol Leu）） in low salinity waters（salinity〈15）,intermediate（1.03–1.25（kg C）/（mol Leu）） in moderate salinity（salinity of 15–25）,and the lowest（0.48–0.85（kg C）/（mol Leu）） in high salinity waters（salinity〉25）.Substrate availability was responsible for spatial variability in the e CFs.In the pristine coastal waters,low e CFs was related to substrate limitation and leucine incorporated was respired to maximize the survival rather than bacterial production.Hence,the e CFs measurement was needed for estimating bacterial production accurately in various marine environments.
文摘Genetic control of leucine content in indica-japonica hybrid rice (Oryza sativa L.) was studied in 35 crosses of F1 and F2 generations, which were derived from crossing 7 male sterile indica rice lines with 5 restorer japonica rice lines along with their parents. Two genetic models and their corresponding statistical methods for quantitative traits of triploid seeds in cereal crops were used for the analysis. The first was the unconditional genetic model, which refers to the analysis of cumulative measurements (from flowering to a specific time) along the developmental stages, while the second was the conditional genetic model, which relates to analysis from one developmental stage to another stage (t - 1→t). The results showed that leucine content of indica-japonica hybrid rice was controlled by the expression of triploid endosperm effect (endosperm additive effect and endosperm dominant effect), cytoplasm effect, diploid maternal plant effect (maternal additive effect and maternal dominant effect) and their environmental interaction effects. Of these effects, endosperm dominant effect and maternal dominant effect were more important at the earlier stages, while endosperm additive effect and maternal additive effect were more important at the later stages of rice grain development under both unconditional and conditional genetic analyses. Due to the high heritabilities, which came from endosperm, maternal and cytoplasm effects for leucine content at different developmental stages, selection for leucine content of indica-japonica hybrid rice would be more efficient at early generations in breeding programs.
文摘Objective: Reactive oxygen species (ROS) are involved in the endothelial-mediated disorders within atherosclerosis. Considering that an oxidant/antioxidant imbalance might be a key factor in the damaging ROS-mediated effects, the present study intends to determine the influence of a high-fat diet, associated with essential amino acids—valine and leucine, upon the experimental animals, through evaluation of plasmatic level of some antioxidant enzymes. Material and Methods: The study was conducted on 32 male Wistar rats, which were fed with cholesterol, valine and leucine, for 60 days. The animals were divided into four groups, according to the received diet: the first group—standard diet;the second group—cholesterol (C);the third group—cholesterol and valine (C + V);the fourth group—cholesterol and leucine (C + L). Evaluations of the oxidative status, through plasma levels of the antioxidant enzymes: superoxide dismutase (SOD) and glutathione peroxidise (GPx), were made for the four mentioned groups of animals, at the beginning of the study (R0), after one (R1) and two months (R2). Results: The average values of SOD and GPx in group of animals fed exclusively with cholesterol (C) were significantly higher compared to the third group where cholesterol was supplemented with valine (C + V) or fourth group fed with cholesterol and leucine (C + L) (p < 0.001), after one month as well at the end of the experiment (two months). There were no significant differences in the levels of SOD and GPx between group III and group IV (p < 0.05) at the end of the experiment. Conclusion: Our results showed that valine and leucine decreased the serum levels of SOD and GPx and therefore they were useful antioxidants, which could improve the endothelial dysfunctions associated with atherosclerosis. Moreover, analysis of the oxidative status in the context of atherosclerotic mediated endothelial damage suggests that deviation from normal to alter endothelial status may be conditioned by an oxidants/antioxidants imbalance.
文摘Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) constitute a family of transcription factors that are characterized by the presence of a DNA-binding domain closely linked with leucine zipper motif functioning in dimer formation. This type of association is unique to plants and considered as an excellent candidate to activate developmental responses to altering environmental conditions. Cotton is the most important fiber plant with a lot of local and commercial uses in the world. HD-Zip proteins not only have key roles in different stages of vascular and inter-fascicular fiber differentiation of cotton but also are suggested to have an important role against abiotic stress that is one of the key factors limiting cotton productivity. Plants have developed various strategies to manage stress conditions through a combination of metabolic, physiological and morphological adaptations. These adaptive changes rely largely on alterations in gene expression. Therefore, transcriptional regulators play a crucial role in stress tolerance. Being a transcription factor HD-Zip might be a useful target for genetic engineering to generate multiple stress tolerance in susceptible plants. In the following chapter, we discussed how the HD-Zip proteins would play a useful role for cotton development both in fiber production and stress adaptation.
基金supported by the National Natural Science Foundation of China(U20A2057)National Basic Research and Development Program of China(2022YFD1300805).
文摘An inadequate amino acid (AA) supply in animals under protein-restricted conditions can slow skeletal muscle growth.Protein translation can be activated by short-term leucine (Leu) stimulation;however,whether muscle mass increases under long-term Leu supplementation and how the gut and muscle respond to Leu supplementation are largely unknown.In this study,we investigated if muscle mass increases with long-term Leu supplementation under protein-restricted conditions.We identified changes in the link between the gut and muscles under different amino acid supply conditions,using goats as the study object.A total of 27 Xiangdong black male goats with average initial body weight (BW)of 10.88±1.22 kg were randomly divided into three dietary treatments:a normal protein diet (NP,14.24%crude protein[CP]);a low protein diet (LP,8.27%CP with supplemental 1.66%rumen-protected lysine[RPLys]and 0.09%rumen-protected methionine[RPMet]);and LP diet with rumen-protected Leu (RPLeu)(LP+RPLeu,8.75%CP with supplemental 1.66%RPLys,0.09%RPMet and 1.46%RPLeu).The animal trial lasted for 110 d,consisting of 20 d of adaptation and a 90 d of experimental period.The results showed that long-term protein restriction increased gut tryptophan hydroxylase 1 (TPH1) activity (P<0.001),tryptophan (Trp) catabolism (P<0.001),and 5-hydroxytryptamine (5-HT) synthesis (P<0.001),which all subsequently reduced goat appetite.Long-term Leu supplementation inhibited 5-HT synthesis(P<0.001),decreased Trp catabolism in the gut,and increased appetite in goats.Long-term protein restriction enhanced jejunal and ileal branched-chain amino acid transferase (BCAT)(P<0.001) and branched-chain a-Keto acid dehydrogenase (BCKD)(P=0.048) activities,which increased branchedchain amino acid (BCAA) catabolism.Immunofluorescence results showed that protein restriction decreased the intestinal mucosal expression of solute carrier family 1 member 5 (SLC1A5)(P=0.032) and solute carrier family 7 member 5 (SLC7A5)(P<0.001),reduced BCAA transport from the mucosa to the blood,lowered BCAA levels in the blood (P<0.001).Western blot results showed that protein restriction inhibited mammalian target of rapamycin (mTOR) pathway activation in goat muscles.Leu supplementation increased BCAA translocation from the intestine to the blood and promoted activation of the muscle m TOR pathway and protein synthesis.In conclusion,our results suggest that Leu supplementation in low-protein diets improves appetite and alleviates the inhibition of muscle protein synthesis in goats.
基金supported in part by the Outsmarting Osteosarcoma Hero Award(Because of Sydney)the UCLA Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research Rose Hill Foundation Innovator Award+23 种基金supported by NCI R01CA201035,R01CA240711,R01CA229893DoD W81XWH-18-1-0223UCLA SPORE in Prostate Cancer(P50 CA092131)JCCC Cancer support grant from NIH P30 CA016042(PI:Teitell)Knut and Alice Wallenberg FoundationBertha Kamprad FoundationDavid H.Koch Prostate Cancer Foundation Young Investigator AwardSwedish Research CouncilSwedish Cancer SocietySIPEA FoundationSwedish Childhood Cancer FoundationJohn and Augusta Perssons FoundationRoyal Physiographic Society of LundFranke and Margareta Bergqvist FoundationCrafoord FoundationLund University Medical Faculty research time allocation award,IngaBrittArne Lundberg Research Foundation,the German Research Foundation(552440240)the German Cancer Consortium(DKTK)the German Federal Ministry of Education and Research(BMBFgrant no.01KD2206A/SATURN3)funding support from the Children’s Discovery Institute of the St.Louis Children’s Hospital.Confocal laser scanning microscopy was performed at the Advanced Light Microscopy/Spectroscopy Laboratory(RRID:SCR_022789)the Leica Microsystems Center of Excellence at the California NanoSystems Institute at UCLA with funding support from NIH Shared Instrumentation Grant S10OD025017Flow cytometry was performed in the UCLA Jonsson Comprehensive Cancer Center(JCCC)Center for AIDS Research Flow Cytometry Core Facility that is supported by National Institutes of Health awards P30 CA016042 and 5P30 AI028697。
文摘Leucine-rich repeat containing 15(LRRC15)has emerged as an attractive biomarker and target for cancer therapy.Transforming growth factor-β(TGFβ)induces the expression of this plasma membrane protein specifically in aggressive and treatment resistant tumor cells derived from mesenchymal stem cells,with minimal expression observed in non-neoplastic tissues.We have developed a humanized monoclonal antibody,DUNP19,that specifically binds with high affinity to a phylogenetically conserved LRRC15 epitope and is rapidly internalized upon LRRC15 binding.In multiple subcutaneous and orthotopic tumor xenograft mouse models,Lutetium-177 labeled DUNP19([^(177)Lu]Lu-DUNP19)enabled non-invasive imaging and molecularly precise radiotherapy to LRRC15-expressing cancer cells and murine cancer-associated fibroblasts,effectively halting tumor progression and prolonging survival with minimal toxicity.Transcriptomic analyses of[^(177)Lu]Lu-DUNP19-treated tumors reveal a loss of pro-tumorigenic mechanisms,including a previously reported TGF β-induced LRRC15+signature associated with immunotherapy resistance.In a syngeneic tumor model,administration of[^(177)Lu]Lu-DUNP19 significantly potentiated checkpoint-blockade therapy,yielding durable complete responses.Together,these results demonstrate that radio-theranostic targeting of LRRC15 with DUNP19 is a compelling precision medicine platform for image-guided diagnosis,eradication,and reprogramming of LRRC15+tumor tissue that drives immunoresistance and disease aggressiveness in a wide range of currently untreatable malignancies.
文摘In this study, ion mobility separation coupled with tandem mass spectrometry （IM-tandem MS） was utilized to investigate the ionization behaviors of two amino acids including leucine and isoleucine. Under the electrospray ionization （ESI） mode, two protonation sites in each molecular sturcture caused two forms of protomer. One arose from the amino being protonated （amino-protomer） and the other from the carboxyl being protonated （car- boxyl-protomer）. In the two-dimensional （drift time, m/z） spectrum, the protomers had the same mass, but the dis- tinguishable drift times and fragmentation patterns. For the characterization purpose, the theoretical collision cross section （CCS） values of the protomers were calculated and proven to be consistent with the experimental. Moreover, the quantified relationship between the amino acids and their protomers was evaluated. It showed that the abun- dance of the carboxyl-protomer was proportional to the concentration of the amino acid, whereas that of the amino- protomer did not have the same trend. Under the atmospheric pressure chemical ionization （APCI） mode, only the carboxyl-protomer was observed. In addition, the amino-protomer and the cluster ions observed under ESI were absent completely. The results demonstrate that the ionization mode impacts heavily on the ionization behaviors of leucine and isoleucine not only on the form of therir protomers but also On the quantified relationship.
基金supported,in part,by a research grant from Singapore Ministry of Education(WBS No.R160-000-664-114).
文摘Soy whey is a liquid by-product generated from tofu production and it is often discarded as wastewater after it is generated.Previous studies have shown that out of the three branched-chain amino acids,isoleucine was found to have an inhibitory effect on Torulaspora delbrueckii Biodiva during soy whey fermentation.In this study,a comparison between isoleucine and leucine was carried out to ascertain how these two structural isomers could affect the growth and metabolism of T.delbrueckii Biodiva at various concentrations(40,80 and 160 mg nitrogen/L).In general,amino acid supplementation sped up the fermentation process relative to unsupplemented samples.Even though glycerol and ethanol production were enhanced in all isoleucine supplemented samples in comparison to unsupplemented samples,isoleucine supplementation resulted in slower yeast growth at day 1 and slower sucrose hydrolysis regardless of the concentration supplemented.As for leucine supplementation,there was no impact on yeast growth,glycerol and ethanol production but sucrose hydrolysis was enhanced in comparison to the unsupplemented samples.Increasing isoleucine and leucine supplementation resulted in corresponding increase in active amyl alcohol and isoamyl alcohol production respectively,but isoleucine had a stimulating effect on increasing the production of other higher alcohols while leucine had an inhibitory effect on the production of other higher alcohols.Therefore,even though isoleucine and leucine are structural isomers with the positional difference of only one methyl group,the impact on the yeast metabolism and growth is visibly different.
基金funded by the Deutsche Forschungsgemeinschaft, an International Reintegration Grant of the European Union, the European Research Council and the German Ministry for Agriculture
文摘The Arabidopsis （Arabidopsis thaliana L.） genome encodes for four distinct classes of homeodomain leucinezipper （HD-ZIP） transcription factors （HD-ZIPI to HD-ZIPIV）, which are all organized in multi-gene families. HD-ZIP transcription factors act as sequence-specific DNA-binding proteins that are able to control the expression level of target genes. While HD-ZIPI and HD-ZIPII proteins are mainly associated with environmental responses, HD-ZIPIII and HD- ZIPIV are primarily known to act as patterning factors. Recent studies have challenged this view. It appears that several of the different HD-ZlP families interact genetically to align both morphogenesis and environmental responses, most likely by modulating phytohormone-signaling networks.
