Validated procedures play an important role to obtain accurate information about individual amino acid requirement data. The aim of the present study was to assess lysine (Lys) requirement of growing chicken both by c...Validated procedures play an important role to obtain accurate information about individual amino acid requirement data. The aim of the present study was to assess lysine (Lys) requirement of growing chicken both by classical supplementation technique and principles of diet dilution technique as applied with “Goettingen approach”. During the starter period (1 - 21 d), a growth study with male meat type chicken (Ross 308) was conducted making use of five graded dietary Lys-levels (3 repetition boxes with 3 birds/box). L-Lys×HCl was gradually added to a diet based on wheat, soybean protein concentrate, wheat gluten and fishmeal to yield 80%, 87.5%, 95%, 102.5% and 110% of the expected requirement level (13 g Lys/kg as fed). Diets were iso-energetic (12.8 MJME/kg) and iso-nitrogenous (21.65% crude protein). Birds were fed on free choice level also to assess the feed intake (FI) effects as important factor on traditional response criteria. Analyzed body composition at start and end of the growth study yielded N deposition (ND) data for further data assessment using exponential approximations depending on dietary Lys content or observed Lys intake. The results indicated significant differences (p < 0.05) in response on body weight gain (BWG) and observed dietary protein quality with unexpected consequences for the derived Lys requirement data. According to the independent variable (Lys in % of diet versus daily Lys intake) and aimed level of daily ND, the needed in-feed content of Lys varied between 1.24% and 1.46%. Application of the exponential modelling by “Goettingen approach” overcame these misleading conclusions by modelling the relationship between required Lys intake and observed response data (BWG, ND) taking also into account the expected real feed intake to formulate the needed in-feed concentration.展开更多
Nitrogen (N) balance studies were conducted with male growing broiler chickens to reevaluate the lysine (Lys) requirement of a modern broiler strain (Ross 308), making use of eight diets with graded crude protein (CP)...Nitrogen (N) balance studies were conducted with male growing broiler chickens to reevaluate the lysine (Lys) requirement of a modern broiler strain (Ross 308), making use of eight diets with graded crude protein (CP) supply (6% - 34% CP as-fed). Wheat, soy protein concentrate, wheat gluten, fishmeal and crystalline amino acids (AAs) were the protein sources in the experimental diets with Lys as limiting AA. Following an adaptation period of five days, two consecutive excreta collection periods (2 × 5 d) were conducted: 10 - 20 d of age (starter period) and 25 - 35 d of age (grower period). Statistical evaluation of N balance data utilized an exponential modelling approach. Based on different dietary Lys efficiency, Lys requirement data were derived by modelling depending on average body weight (BW) during starter and grower period and targeted body protein deposition (PD), respectively. In addition, the influence of graded feed intake was taken into account. For the starter period at 600 g BW and assumed 10 g daily body PD, Lys requirement data between 741 mg and 823 mg per day were observed. The corresponding Lys in-feed concentration was 1.06% and 1.18%, dependent on supposed Lys efficiency at 70 g daily feed intake. For the grower period (average BW 1800 g), 1272 mg to 1473 mg Lys per day was needed to yield 16.5 g daily PD. The corresponding required Lys in-feed concentration was between 0.85% and 0.94% Lys for 150 g daily feed intake.展开更多
Optimal dietary methionine (Met) to lysine (Lys) ratio in presence of elevated dietary cysteine (Cys) levels was derived for meat type growing chicken. Twelve averaged weighed Ross 308 birds (each 50% of male and fema...Optimal dietary methionine (Met) to lysine (Lys) ratio in presence of elevated dietary cysteine (Cys) levels was derived for meat type growing chicken. Twelve averaged weighed Ross 308 birds (each 50% of male and female per dietary treatment) were utilized in N balance trials. During starter (d10 - 20) and grower period (d25 - 35) five dietary treatments were used. Diets based on uniform mixtures of maize, wheat, soybean meal, potato protein and fish meal were supplemented with crystalline amino acids (AA). In diets 1 - 3, the dietary Cys to Met ratio was set as 85, 95 and 105 to 100, respectively. Diet 4, at a Cys to Met ratio of 105 to 100, was additionally supplemented with betaine (BET) as methyl group donor. Diets 1 - 4 were limiting in Met, diet 5 without L-Lys·HCl addition was limiting in Lys. Individual N-balance data per treatment were utilized for assessing protein quality and efficiency of dietary Met (Diets 1 - 4) or Lys (Diet 5) based on “Goettingen approach”. Elevated dietary Cys supply and supplemented BET failed to improve both dietary protein quality and Met efficiency. The established optimal Met to Lys ratio was on average 34 to 100 for growing chicken during starter and grower period, respectively.展开更多
BACKGROUND Myocardial remodeling is a key factor in the progression of cardiovascular disease to the end stage.In addition to myocardial infarction or stress overload,dietary factors have recently been considered asso...BACKGROUND Myocardial remodeling is a key factor in the progression of cardiovascular disease to the end stage.In addition to myocardial infarction or stress overload,dietary factors have recently been considered associated with myocardial remodeling.Nε-(carboxymethyl)lysine(CML)is a representative foodborne toxic product,which can be ingested via daily diet.Therefore,there is a marked need to explore the effects of dietary CML on the myocardium.AIM To explore the effects of dietary CML(dCML)on the heart.METHODS C57 BL/6 mice were divided into a control group and a dCML group.The control group and the dCML group were respectively fed a normal diet or diet supplemented with CML for 20 wk.Body weight and blood glucose were recorded every 4 wk.^(18)F-fluorodeoxyglucose(FDG)was used to trace the glucose uptake in mouse myocardium,followed by visualizing with micro-positron emission tomography(PET).Myocardial remodeling and glucose metabolism were also detected.In vitro,H9C2 cardiomyocytes were added to exogenous CML and cultured for 24 h.The effects of exogenous CML on glucose metabolism,collagen I expression,hypertrophy,and apoptosis of cardiomyocytes were analyzed.RESULTS Our results suggest that the levels of fasting blood glucose,fasting insulin,and serum CML were significantly increased after 20 wk of dCML.Micro-PET showed that ^(18)F-FDG accumulated more in the myocardium of the dCML group than in the control group.Histological staining revealed that dCML could lead to myocardial fibrosis and hypertrophy.The indexes of myocardial fibrosis,apoptosis,and hypertrophy were also increased in the dCML group,whereas the activities of glucose metabolism-related pathways and citrate synthase(CS)were significantly inhibited.In cardiomyocytes,collagen I expression and cellular size were significantly increased after the addition of exogenous CML.CML significantly promoted cellular hypertrophy and apoptosis,while pathways involved in glucose metabolism and level of Cs mRNA were significantly inhibited.CONCLUSION This study reveals that dCML alters myocardial glucose metabolism and promotes myocardial remodeling.展开更多
BACKGROUND Advanced glycation end products(AGEs)are diabetic metabolic toxic products that cannot be ignored.Nε-(carboxymethyl)lysine(CML),a component of AGEs,could increase macrophage lipid uptake,promote foam cell ...BACKGROUND Advanced glycation end products(AGEs)are diabetic metabolic toxic products that cannot be ignored.Nε-(carboxymethyl)lysine(CML),a component of AGEs,could increase macrophage lipid uptake,promote foam cell formation,and thereby accelerate atherosclerosis.The receptor for AGEs(RAGE)and cluster of differentiation 36(CD36)were the receptors of CML.However,it is still unknown whether RAGE and CD36 play key roles in CML-promoted lipid uptake.AIM Our study aimed to explore the role of RAGE and CD36 in CML-induced macrophage lipid uptake.METHODS In this study,we examined the effect of CML on lipid uptake by Raw264.7 macrophages.After adding 10 mmol/L CML,the lipid accumulation in macrophages was confirmed by oil red O staining.Expression changes of CD36 and RAGE were detected with immunoblotting and quantitative real-time polymerase chain reaction.The interaction between CML with CD36 and RAGE was verified by immunoprecipitation.We synthesized a novel N-succinimidyl-4-18Ffluorobenzoate-CML radioactive probe.Radioactive receptor-ligand binding assays were performed to test the binding affinity between CML with CD36 and RAGE.The effects of blocking CD36 or RAGE on CML-promoting lipid uptake were also detected.RESULTS The study revealed that CML significantly promoted lipid uptake by macrophages.Immunoprecipitation and radioactive receptor-ligand binding assays indicated that CML could specifically bind to both CD36 and RAGE.CML had a higher affinity for CD36 than RAGE.ARG82,ASN71,and THR70 were the potential interacting amino acids that CD36 binds to CML Anti-CD36 and anti-RAGE could block the uptake of CML by macrophages.The lipid uptake promotion effect of CML was significantly attenuated after blocking CD36 or RAGE.CONCLUSION Our results suggest that the binding of CML with CD36 and RAGE promotes macrophage lipid uptake.展开更多
Amphiphilic block copolymers poly(LysAA-b-DMS) consisting of a hydrophilic poly(N-α-acrylamide-L-lysine) [poly(LysAA)] segment with different molecular weights and a hydrophobic polydimethylsiloxane (PDMS) segment we...Amphiphilic block copolymers poly(LysAA-b-DMS) consisting of a hydrophilic poly(N-α-acrylamide-L-lysine) [poly(LysAA)] segment with different molecular weights and a hydrophobic polydimethylsiloxane (PDMS) segment were prepared as follows. The precursor copolymer poly(Boc-LysAA-OtBu-b-PDMS) was obtained from radical polymerization of N-α-acrylamide-N-ε-tert-butoxycarbonyl-L-lysine-tert-butylester (Boc-LysAA-OtBu) initiated with 4,4’-azobis(polydimethylsiloxane 4-cyanopentanoate) (azo-PDMS) with the molecular weight of PDMS Mw = 4.3 × 103 in the presence of 2-mercaptoethanol (2-ME) as a chain-transfer agent. Removal of the protecting groups of the precursor copolymer was carried out in 80% trifluoroacetic acid aqueous solution to give poly(LysAA-b-DMS)-1-3. The weight average molecular weight of poly(LysAA-b-DMS)-1-3 was Mw = 1.02 × 104 – 2.52 × 104. From the 1H-NMR and fluorescence spectra measurements, poly(LysAA-b-DMS)-1-3 was determined to self-organize and form core-shell micelles in water. The critical micelle concentration (CMC) increased to 1000 - 4000 mg·L–1 with increasing molar ratio of the poly(LysAA) segment from 0.42 to 0.65. From morphological analysis with a scanning probe microscope (SPM), poly(LysAA-b-DMS) has microphase-separated structures made up of hydrophilic and hydrophobic regions with the domain size ranging from several tens to several hundreds of nanometers. Inhibition of thrombin activity of poly(LysAA-b-DMS) was evaluated from the Michaelis constant (KM) and catalytic activity (kcat) for the enzymatic reaction of thrombin and synthetic substrate S-2238 in the presence of poly(LysAA-b-DMS). The KM and kcat were 0.10 - 0.11 mM and 4.04 × 105 – 4.26 × 105 min–1, respectively. Fibrinolytic activity was also verified from the transformation of plasminogen to plasmin by tissue plasminogen activator (t-PA) using synthetic substrate S-2251 in the presence of poly(LysAA-b-DMS). The KM and kcat were 0.07 mM and 5.73 × 106 –5.95 × 106 min–1, respectively.展开更多
Our previous study showed an association between advanced glycation end products (AGEs) and neural tube defects (NTDs). To understand the molecular mechanisms underlying the effect of AGEs on neural tube developme...Our previous study showed an association between advanced glycation end products (AGEs) and neural tube defects (NTDs). To understand the molecular mechanisms underlying the effect of AGEs on neural tube development, C57BL/6 female mice were fed for 4 weeks with com- mercial food containing 3% advanced glycation end product bovine serum albumin (AGE-BSA) or 3% bovine serum albumin (BSA) as a control. After mating mice, oxidative stress markers including malondialdehyde and H202 were measured at embryonic day 7.5 (E7.5) of ges- tation, and the level of intracellular reactive oxygen species (ROS) in embryonic cells was determined at E8.5. In addition to evaluating NTDs, an enzyme-linked immunosorbent assay was used to determine the effect of embryonic protein administration on the N-(carboxymethyl) lysine reactivity of acid and carboxyethyl lysine antibodies at E10.5. The results showed a remarkable increase in the incidence of NTDs at El0.5 in embryos of mice fed with AGE-BSA (no hyperglycemia) compared with control mice. Moreover, embryonic protein administration resulted in a noticeable increase in the reactivity of N-(carboxymethyl) lysine and N(ε)-(carboxyethyl) lysine antibodies. Malondialdehyde and H2O2 levels in embryonic cells were increased at E7.5, followed by increased intracellular ROS levels at E8.5. Vitamin E supplementation could partially recover these phenomena. Collectively, these results suggest that AGE-BSA could induce NTDs in the absence of hyperglycemia by an underlying mechanism that is at least partially associated with its capacity to increase embryonic oxidative stress levels.展开更多
文摘Validated procedures play an important role to obtain accurate information about individual amino acid requirement data. The aim of the present study was to assess lysine (Lys) requirement of growing chicken both by classical supplementation technique and principles of diet dilution technique as applied with “Goettingen approach”. During the starter period (1 - 21 d), a growth study with male meat type chicken (Ross 308) was conducted making use of five graded dietary Lys-levels (3 repetition boxes with 3 birds/box). L-Lys×HCl was gradually added to a diet based on wheat, soybean protein concentrate, wheat gluten and fishmeal to yield 80%, 87.5%, 95%, 102.5% and 110% of the expected requirement level (13 g Lys/kg as fed). Diets were iso-energetic (12.8 MJME/kg) and iso-nitrogenous (21.65% crude protein). Birds were fed on free choice level also to assess the feed intake (FI) effects as important factor on traditional response criteria. Analyzed body composition at start and end of the growth study yielded N deposition (ND) data for further data assessment using exponential approximations depending on dietary Lys content or observed Lys intake. The results indicated significant differences (p < 0.05) in response on body weight gain (BWG) and observed dietary protein quality with unexpected consequences for the derived Lys requirement data. According to the independent variable (Lys in % of diet versus daily Lys intake) and aimed level of daily ND, the needed in-feed content of Lys varied between 1.24% and 1.46%. Application of the exponential modelling by “Goettingen approach” overcame these misleading conclusions by modelling the relationship between required Lys intake and observed response data (BWG, ND) taking also into account the expected real feed intake to formulate the needed in-feed concentration.
文摘Nitrogen (N) balance studies were conducted with male growing broiler chickens to reevaluate the lysine (Lys) requirement of a modern broiler strain (Ross 308), making use of eight diets with graded crude protein (CP) supply (6% - 34% CP as-fed). Wheat, soy protein concentrate, wheat gluten, fishmeal and crystalline amino acids (AAs) were the protein sources in the experimental diets with Lys as limiting AA. Following an adaptation period of five days, two consecutive excreta collection periods (2 × 5 d) were conducted: 10 - 20 d of age (starter period) and 25 - 35 d of age (grower period). Statistical evaluation of N balance data utilized an exponential modelling approach. Based on different dietary Lys efficiency, Lys requirement data were derived by modelling depending on average body weight (BW) during starter and grower period and targeted body protein deposition (PD), respectively. In addition, the influence of graded feed intake was taken into account. For the starter period at 600 g BW and assumed 10 g daily body PD, Lys requirement data between 741 mg and 823 mg per day were observed. The corresponding Lys in-feed concentration was 1.06% and 1.18%, dependent on supposed Lys efficiency at 70 g daily feed intake. For the grower period (average BW 1800 g), 1272 mg to 1473 mg Lys per day was needed to yield 16.5 g daily PD. The corresponding required Lys in-feed concentration was between 0.85% and 0.94% Lys for 150 g daily feed intake.
文摘Optimal dietary methionine (Met) to lysine (Lys) ratio in presence of elevated dietary cysteine (Cys) levels was derived for meat type growing chicken. Twelve averaged weighed Ross 308 birds (each 50% of male and female per dietary treatment) were utilized in N balance trials. During starter (d10 - 20) and grower period (d25 - 35) five dietary treatments were used. Diets based on uniform mixtures of maize, wheat, soybean meal, potato protein and fish meal were supplemented with crystalline amino acids (AA). In diets 1 - 3, the dietary Cys to Met ratio was set as 85, 95 and 105 to 100, respectively. Diet 4, at a Cys to Met ratio of 105 to 100, was additionally supplemented with betaine (BET) as methyl group donor. Diets 1 - 4 were limiting in Met, diet 5 without L-Lys·HCl addition was limiting in Lys. Individual N-balance data per treatment were utilized for assessing protein quality and efficiency of dietary Met (Diets 1 - 4) or Lys (Diet 5) based on “Goettingen approach”. Elevated dietary Cys supply and supplemented BET failed to improve both dietary protein quality and Met efficiency. The established optimal Met to Lys ratio was on average 34 to 100 for growing chicken during starter and grower period, respectively.
基金Supported by the National Natural Science Foundation of China,No.82070455Natural Science Foundation of Jiangsu Province,No.BK20201225+1 种基金Medical Innovation Team Project of Jiangsu Province,No.CXTDA2017010Research and Innovation Funding Project for College Students in Experimental Animal Center of Jiangsu University。
文摘BACKGROUND Myocardial remodeling is a key factor in the progression of cardiovascular disease to the end stage.In addition to myocardial infarction or stress overload,dietary factors have recently been considered associated with myocardial remodeling.Nε-(carboxymethyl)lysine(CML)is a representative foodborne toxic product,which can be ingested via daily diet.Therefore,there is a marked need to explore the effects of dietary CML on the myocardium.AIM To explore the effects of dietary CML(dCML)on the heart.METHODS C57 BL/6 mice were divided into a control group and a dCML group.The control group and the dCML group were respectively fed a normal diet or diet supplemented with CML for 20 wk.Body weight and blood glucose were recorded every 4 wk.^(18)F-fluorodeoxyglucose(FDG)was used to trace the glucose uptake in mouse myocardium,followed by visualizing with micro-positron emission tomography(PET).Myocardial remodeling and glucose metabolism were also detected.In vitro,H9C2 cardiomyocytes were added to exogenous CML and cultured for 24 h.The effects of exogenous CML on glucose metabolism,collagen I expression,hypertrophy,and apoptosis of cardiomyocytes were analyzed.RESULTS Our results suggest that the levels of fasting blood glucose,fasting insulin,and serum CML were significantly increased after 20 wk of dCML.Micro-PET showed that ^(18)F-FDG accumulated more in the myocardium of the dCML group than in the control group.Histological staining revealed that dCML could lead to myocardial fibrosis and hypertrophy.The indexes of myocardial fibrosis,apoptosis,and hypertrophy were also increased in the dCML group,whereas the activities of glucose metabolism-related pathways and citrate synthase(CS)were significantly inhibited.In cardiomyocytes,collagen I expression and cellular size were significantly increased after the addition of exogenous CML.CML significantly promoted cellular hypertrophy and apoptosis,while pathways involved in glucose metabolism and level of Cs mRNA were significantly inhibited.CONCLUSION This study reveals that dCML alters myocardial glucose metabolism and promotes myocardial remodeling.
基金Supported by The National Natural Science Foundation of China,No.82070455Natural Science Foundation of Jiangsu Province,No.BK20201225Medical Innovation Team Project of Jiangsu Province,No.CXTDA2017010。
文摘BACKGROUND Advanced glycation end products(AGEs)are diabetic metabolic toxic products that cannot be ignored.Nε-(carboxymethyl)lysine(CML),a component of AGEs,could increase macrophage lipid uptake,promote foam cell formation,and thereby accelerate atherosclerosis.The receptor for AGEs(RAGE)and cluster of differentiation 36(CD36)were the receptors of CML.However,it is still unknown whether RAGE and CD36 play key roles in CML-promoted lipid uptake.AIM Our study aimed to explore the role of RAGE and CD36 in CML-induced macrophage lipid uptake.METHODS In this study,we examined the effect of CML on lipid uptake by Raw264.7 macrophages.After adding 10 mmol/L CML,the lipid accumulation in macrophages was confirmed by oil red O staining.Expression changes of CD36 and RAGE were detected with immunoblotting and quantitative real-time polymerase chain reaction.The interaction between CML with CD36 and RAGE was verified by immunoprecipitation.We synthesized a novel N-succinimidyl-4-18Ffluorobenzoate-CML radioactive probe.Radioactive receptor-ligand binding assays were performed to test the binding affinity between CML with CD36 and RAGE.The effects of blocking CD36 or RAGE on CML-promoting lipid uptake were also detected.RESULTS The study revealed that CML significantly promoted lipid uptake by macrophages.Immunoprecipitation and radioactive receptor-ligand binding assays indicated that CML could specifically bind to both CD36 and RAGE.CML had a higher affinity for CD36 than RAGE.ARG82,ASN71,and THR70 were the potential interacting amino acids that CD36 binds to CML Anti-CD36 and anti-RAGE could block the uptake of CML by macrophages.The lipid uptake promotion effect of CML was significantly attenuated after blocking CD36 or RAGE.CONCLUSION Our results suggest that the binding of CML with CD36 and RAGE promotes macrophage lipid uptake.
文摘Amphiphilic block copolymers poly(LysAA-b-DMS) consisting of a hydrophilic poly(N-α-acrylamide-L-lysine) [poly(LysAA)] segment with different molecular weights and a hydrophobic polydimethylsiloxane (PDMS) segment were prepared as follows. The precursor copolymer poly(Boc-LysAA-OtBu-b-PDMS) was obtained from radical polymerization of N-α-acrylamide-N-ε-tert-butoxycarbonyl-L-lysine-tert-butylester (Boc-LysAA-OtBu) initiated with 4,4’-azobis(polydimethylsiloxane 4-cyanopentanoate) (azo-PDMS) with the molecular weight of PDMS Mw = 4.3 × 103 in the presence of 2-mercaptoethanol (2-ME) as a chain-transfer agent. Removal of the protecting groups of the precursor copolymer was carried out in 80% trifluoroacetic acid aqueous solution to give poly(LysAA-b-DMS)-1-3. The weight average molecular weight of poly(LysAA-b-DMS)-1-3 was Mw = 1.02 × 104 – 2.52 × 104. From the 1H-NMR and fluorescence spectra measurements, poly(LysAA-b-DMS)-1-3 was determined to self-organize and form core-shell micelles in water. The critical micelle concentration (CMC) increased to 1000 - 4000 mg·L–1 with increasing molar ratio of the poly(LysAA) segment from 0.42 to 0.65. From morphological analysis with a scanning probe microscope (SPM), poly(LysAA-b-DMS) has microphase-separated structures made up of hydrophilic and hydrophobic regions with the domain size ranging from several tens to several hundreds of nanometers. Inhibition of thrombin activity of poly(LysAA-b-DMS) was evaluated from the Michaelis constant (KM) and catalytic activity (kcat) for the enzymatic reaction of thrombin and synthetic substrate S-2238 in the presence of poly(LysAA-b-DMS). The KM and kcat were 0.10 - 0.11 mM and 4.04 × 105 – 4.26 × 105 min–1, respectively. Fibrinolytic activity was also verified from the transformation of plasminogen to plasmin by tissue plasminogen activator (t-PA) using synthetic substrate S-2251 in the presence of poly(LysAA-b-DMS). The KM and kcat were 0.07 mM and 5.73 × 106 –5.95 × 106 min–1, respectively.
基金supported by the grant from Shaanxi Technology Committee of China,No.2013JM4001the China Scholarship Council(CSC)
文摘Our previous study showed an association between advanced glycation end products (AGEs) and neural tube defects (NTDs). To understand the molecular mechanisms underlying the effect of AGEs on neural tube development, C57BL/6 female mice were fed for 4 weeks with com- mercial food containing 3% advanced glycation end product bovine serum albumin (AGE-BSA) or 3% bovine serum albumin (BSA) as a control. After mating mice, oxidative stress markers including malondialdehyde and H202 were measured at embryonic day 7.5 (E7.5) of ges- tation, and the level of intracellular reactive oxygen species (ROS) in embryonic cells was determined at E8.5. In addition to evaluating NTDs, an enzyme-linked immunosorbent assay was used to determine the effect of embryonic protein administration on the N-(carboxymethyl) lysine reactivity of acid and carboxyethyl lysine antibodies at E10.5. The results showed a remarkable increase in the incidence of NTDs at El0.5 in embryos of mice fed with AGE-BSA (no hyperglycemia) compared with control mice. Moreover, embryonic protein administration resulted in a noticeable increase in the reactivity of N-(carboxymethyl) lysine and N(ε)-(carboxyethyl) lysine antibodies. Malondialdehyde and H2O2 levels in embryonic cells were increased at E7.5, followed by increased intracellular ROS levels at E8.5. Vitamin E supplementation could partially recover these phenomena. Collectively, these results suggest that AGE-BSA could induce NTDs in the absence of hyperglycemia by an underlying mechanism that is at least partially associated with its capacity to increase embryonic oxidative stress levels.