Effects of different energy levels in low-protein diet on liver lipid metabolism in the late-phase laying hens through the gut-liver axis

Background The energy/protein imbalance in a low-protein diet induces lipid metabolism disorders in late-phase laying hens.Reducing energy levels in the low-protein diet to adjust the energy-to-protein ratio may improve fat deposition,but this also decreases the laying performance of hens.This study investigated the mechanism by which different energy levels in the low-protein diet influences liver lipid metabolism in late-phase laying hens through the enterohepatic axis to guide feed optimization and nutrition strategies.A total of 288 laying hens were randomly allocated to the normal-energy and normal-protein diet group(positive control:CK)or 1 of 3 groups:lowenergy and low-protein diet(LL),normal-energy and low-protein diet(NL),and high-energy and low-protein diet(HL)groups.The energy-to-protein ratios of the CK,LL,NL,and HL diets were 0.67,0.74,0.77,and 0.80,respectively.Results Compared with the CK group,egg quality deteriorated with increasing energy intake in late-phase laying hens fed low-protein diet.Hens fed LL,NL,and HL diets had significantly higher triglyceride,total cholesterol,acetylCo A carboxylase,and fatty acid synthase levels,but significantly lower hepatic lipase levels compared with the CK group.Liver transcriptome sequencing revealed that genes involved in fatty acid beta-oxidation(ACOX1,HADHA,EHHADH,and ACAA1)were downregulated,whereas genes related to fatty acid synthesis(SCD,FASN,and ACACA)were upregulated in LL group compared with the CK group.Comparison of the cecal microbiome showed that in hens fed an LL diet,Lactobacillus and Desulfovibrio were enriched,whereas riboflavin metabolism was suppressed.Cecal metabolites that were most significantly affected by the LL diet included several vitamins,such as riboflavin(vitamin B2),pantethine(vitamin B5 derivative),pyridoxine(vitamin B6),and 4-pyridoxic acid.Conclusion A lipid metabolism disorder due to deficiencies of vitamin B2 and pantethine originating from the metabolism of the cecal microbiome may be the underlying reason for fat accumulation in the liver of late-phase laying hens fed an LL diet.Based on the present study,we propose that targeting vitamin B2 and pantethine(vitamin B5 derivative)might be an effective strategy for improving lipid metabolism in late-phase laying hens fed a low-protein diet.

Effects of Different Protein Energy Ratios on Fattening Performance,Slaughter Performance and Meat Quality of Pigs

[Objectives]Protein energy ratio refers to the proportional relationship between protein and energy levels in animal diets,i.e.,the grams of crude protein corresponding to every megacalorie of energy,which is generally expressed as CP:ME or CP:DE.This study was conducted to investigate the effects of different diets on fattening and slaughter performance and meat quality traits for"L(Large Yorkshire)×L(Landrace)"crossbred pigs.[Methods]Eighteen piglets of L×L crossbred with similar body weights about 51 kg were selected.The piglets were divided into 3 groups randomly and each group was assigned to 3 replicates with 2 piglets in a replicate.Group A was fed diet Ⅰ (control diet),group B was fed the same diet of group A in the first month of the trial but fed diet Ⅱ in second month,and group C consumed diet Ⅲ.All the pigs were fed in the same feeding condition for two months except the different diets during the trial.One pig in similar body weight from each replicate was slaughtered for the determination of slaughter performance and meat quality traits in the end of the trial.[Results]The average daily feed intake(ADFI)of pigs from group B was only(2.32±0.52)kg and significantly lower than those from group A and group C(P<0.05),but no difference was found in average daily gain(ADG) and feed/gain(P>0.05).Also,no differences occurred in the carcass length,back fat thickness,longissimus muscle (LM) area and dressing percentage(P>0.05).In addition,no differences were found in the meat quality traits of shear force,meat color,pH_(45 min),pH_(24 h) and cooking loss(P>0.05).However,the water-holding capacity of meat from group C was(2.58±0.02)ms and significantly lower than that of(2.80±0.20)ms from group A(P<0.05).Although the contents of glutamic acid and cystine in LM from group B was significantly lower than those from group A and group C(P<0.05),no differences occurred in the contents of other amino acids,the total amino acid and total flavor amino acid among the three groups(P>0.05).However,the inosine monophosphate content of LM from group C was only(331.80±11.53)mg/100 g and significantly lower than those of(361.00±6.36)and(366.37±4.80)mg/100 g from group A and B(P<0.05).Even though no differences were found in the contents of DM and CP in LM among the three groups of pigs,the content of intramuscular fat(IMF)in LM from group B and group C was increased by 45.6%and 46.58%respectively from that of group A(P<0.05),but no difference occurred between group B and C(P>0.05).[Conclusions]DietsⅡ and Ⅲ in this study caused no differences in fattening and slaughter performance of L×L crossbred pigs,but the effects on some meat traits were still significant,especially on the improvement of intramuscular fat in experimental pigs.Therefore,they could improve the meat quality of crossbred pigs to a certain extent.

Effects of Astragalus membranaceus on Energy Metabolism and Expression of CNTF Protein in Skeletal Muscle of Exercise-induced Fatigue Rats

[Objectives]This study was conducted to investigate the effects of Astragalus membranaceus in different groups on energy metabolism and CNTF protein expression in skeletal muscle of exercise-induced fatigue rats.[Methods]Thirty-five clean male SD rats were randomly divided into a normal group,and low-,meddle-and high-dose groups of A.membranaceus aqueous solution,with 7 rats in each group.The low-dose,medium-dose and high-dose groups were given by gavage at 0.65,1.3 and 2.6 g/kg,respectively,while the normal group and the model group were given normal food and water.The weight of rats was observed.The contents of serum urea,lactate,muscle glycogen,liver glycogen and CNTF expression were detected.[Results]After modeling,compared with the normal group,the serum lactate and urea contents of rats in the model group significantly increased(P<0.01),while the muscle glycogen content(P<0.01)and liver glycogen content(P<0.05)of the skeletal muscle significantly decreased.Compared with the model group,the low-,meddle-and high-dose groups of A.membranaceus significantly reduced the levels of lactate and urea in serum(P<0.01),while the levels of muscle glycogen and liver glycogen in the skeletal muscle significantly increased(P<0.01,P<0.05).[Conclusions]This study provides a good research foundation for the treatment of exercise-induced fatigue using traditional Chinese herb A.membranaceus in modern clinical practice.

Targeted metabolomics reveals the aberrant energy status in diabetic peripheral neuropathy and the neuroprotective mechanism of traditional Chinese medicine JinMaiTong

Diabetic peripheral neuropathy (DPN) is a common and devastating complication of diabetes, for which effective therapies are currently lacking. Disturbed energy status plays a crucial role in DPN pathogenesis. However, the integrated profile of energy metabolism, especially the central carbohydrate metabolism, remains unclear in DPN. Here, we developed a metabolomics approach by targeting 56 metabolites using high-performance ion chromatography-tandem mass spectrometry (HPIC-MS/MS) to illustrate the integrative characteristics of central carbohydrate metabolism in patients with DPN and streptozotocin-induced DPN rats. Furthermore, JinMaiTong (JMT), a traditional Chinese medicine (TCM) formula, was found to be effective for DPN, improving the peripheral neurological function and alleviating the neuropathology of DPN rats even after demyelination and axonal degeneration. JMT ameliorated DPN by regulating the aberrant energy balance and mitochondrial functions, including excessive glycolysis restoration, tricarboxylic acid cycle improvement, and increased adenosine triphosphate (ATP) generation. Bioenergetic profile was aberrant in cultured rat Schwann cells under high-glucose conditions, which was remarkably corrected by JMT treatment. In-vivo and in-vitro studies revealed that these effects of JMT were mainly attributed to the activation of adenosine monophosphate (AMP)-activated protein kinase (AMPK) and downstream peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α). Our results expand the therapeutic framework for DPN and suggest the integrative modulation of energy metabolism using TCMs, such as JMT, as an effective strategy for its treatment.

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

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

Estimation of Digestible Energy Values of Plant Protein Supplement in Pig

[Objective] The aim of the study was to establish the effective and accurate formulas for estimating the digestible energy （DE） values of plant protein supplement in pig. [Method] By difference method with different amount of alternative feeds （20% -50%）, two4 x4 Latin- square-designed trials were taken on eight castrated male pigs [ Yorkshire x Landrace x Neijiang pig, initial body-weight： （46 ±2） kg ] to deter- mine the apparent digestible energy （ADE） of the eight kinds of plant protein supplement commonly used in China, that is, corn gluten meal （sol.）, soybean meal （ sol. ）, fababean, pea, rapeseed meal （ sol. ）, sesame meal （ sol. ）, rapeseed meal （ exp. ） and cotton seed meal （sol.）. [Resultl （1） Fiber was the most important factor to estimate the ADE of plant protein supplement in pigs, and ADF was the best one. （2） The most effective equations were as below： （ 1 ） OE （kJ/kg DM） = 14 741.86 - 185.01ADF＋54.01SCHO＋22.45CP （ R =0.988,RSD= 67.9,P〈0.01 ） ; （2） DE （kJ/kg DM） =22 223.26 -209.58ADF＋26.79SCHO-1.09GE （ Ff =0.989,RSD=66.9, P〈0.01 ） . [Conclusion] The accurate, practical and specific regression equations were established for DE prediction of plant protein supplement in pig.

Effects of dietary protein/energy ratio on growth performance,carcass trait,meat quality,and plasma metabolites in pigs of different genotypes

Background： The protein/energy ratio is important for the production performance and utilization of available feed resources by animals. Increased protein consumption by mammals leads to elevated feed costs and increased nitrogen release into the environment. This study aimed to evaluate the effects of dietary protein/energy ratio on the growth performance, carcass traits, meat quality, and plasma metabolites of pigs of different genotypes. Methods： Bama mini-pigs and Landrace pigs were randomly assigned to two dietary treatment groups （Chinese conventional diet with low protein/energy ratio or National Research Council diet with high protein/energy ratio; n = 24 per treatment） in a 2 x 2 factorial arrangement. Blood and muscle samples were collected at the end of the nursery, growing, and finishing phases. Results： We observed significant interactions （P 〈 0.05） between breed and diet for total fat percentage, intramuscular fat （IMF） content, protein content in biceps femoris （BF） muscle, and plasma urea nitrogen （UN） concentration in the nursery phase; for average daily gain （ADG）, average daily feed intake （ADFI）, dry matter, IMF content in psoas major （PM） muscle, and plasma total protein and albumin concentrations in the growing phase; and for drip loss and plasma UN concentration in the finishing phase. Breed influenced （P 〈 0.05） growth performance, carcass traits, and meat quality, but not plasma metabolites. Throughout the trial, Landrace pigs showed significantly higher （P 〈 0.0_5） ADG, ADFI, dressing percentage, lean mass rate, and loin-eye area than did Bama mini-pigs, but significantly lower （P 〈 0.0.5） feed/gain ratio, fat percentage, backfat thickness, and IMF content. Dietary protein/energy ratio influenced the pH value, chemical composition of BF and PM muscles, and plasma activities of glutamic-pyruvic transaminase and gamma-glutamyl transpeptidase, and plasma concentration of UN. Conclusions： Compared with Landrace pigs, Bama mini-pigs showed slower growth and lower carcass performance, but had better meat quality. Moreover, unlike Landrace pigs, the dietary protein/energy ratio did not affect the growth performance of Bama mini-pigs. These results suggest that, in swine production, low dietary protein/energy ratio may be useful for reducing feed costs and minimizing the adverse effects of ammonia release into the environment.

Energy and protein requirements for maintenance of Hu sheep during pregnancy

This study aimed to determine the effect of stage and level of feed intake on energy metabolism, carbon-nitrogen （C-N） balance, and methane emission to determine energy and protein requirements for maintenance of maternal body including pregnancy tissues during pregnancy using the method of C-N balance. Twenty-one ewes carrying twin fetuses were randomly divided into three groups of seven ewes each in the digestion and respirometry trial at d 40, 100, and 130 of gestation, respectively. Three groups were fed a mixed diet either for ad lib#urn intake, 70 or 50% of the ad libfum intake during pregnancy. The results showed that the apparent digestibility of C and N were increased as feeding levels decreased at each stage of gestation. The daily net energy requirements for maintenance （NEro） were 295.80, 310.09, and 323.59 kJ kg-1 BW0.75 （metabolic body weight） with a partial efficiency of metabolisable energy utilization for maintenance of 0.664, 0.644, and 0.620 at d 40, 100, and 130 of gestation, respectively. The daily net protein requirements for maintenance were 1.99, 2.35, and 2.99 g kg-1 BW0.75 at d 40, 100, and 130 of gestation, respectively. These results for the nutritional requirements of the net energy and protein may help to formulate more balanced diets for Hu sheep during pregnancy.

Alternative prediction methods of protein and energy evaluation of pig feeds

Precise knowledge of the actual nutritional value of individual feedstuffs and complete diets for pigs is important for efficient livestock production. Methods of assessment of protein and energy values in pig feeds have been briefly described. In vivo determination of protein and energy values of feeds in pigs are time-consuming,expensive and very often require the use of surgically-modified animals. There is a need for more simple, rapid,inexpensive and reproducible methods for routine feed evaluation. Protein and energy values of pig feeds can be estimated using the following alternative methods: 1) prediction equations based on chemical composition; 2)animal models as rats, cockerels and growing pigs for adult animals; 3) rapid methods, such as the mobile nylon bag technique and in vitro methods. Alternative methods developed for predicting the total tract and ileal digestibility of nutrients including amino acids in feedstuffs and diets for pigs have been reviewed. This article focuses on two in vitro methods that can be used for the routine evaluation of amino acid ileal digestibility and energy value of pig feeds and on factors affecting digestibility determined in vivo in pigs and by alternative methods. Validation of alternative methods has been carried out by comparing the results obtained using these methods with those acquired in vivo in pigs. In conclusion, energy and protein values of pig feeds may be estimated with satisfactory precision in rats and by the two-or three-step in vitro methods providing equations for the calculation of standardized ileal digestibility of amino acids and metabolizable energy content. The use of alternative methods of feed evaluation is an important way for reduction of stressful animal experiments.

Studies on Reducing Nitrogen Excretion: Ⅰ. Net Energy Requirement of Finishing Pigs Maximizing Performance and Carcass Quality Fed Low Crude Protein Diets Supplemented with Crystalline Amino Acids

Two experiments were conducted to in- vestigate the effects of net energy （NE） level on the performance and carcass traits of finishing pigs fed low crude protein （CP） diets supplemented with crystalline amino acids （CAA）. A total of 216 （Exp. 1） and 360 （Exp. 2 ） barrows （ Yorkshire × Landrace× Duroc ） were allotted to one of six treatments （n =6）. The experimental diets were based on corn and soybean meal, and consisted of a high-CP diet that contained approximately 16% CP and 2.50 Mcal/kg of NE as well as five low-CP diets in which the CP level of the diet was reduced by approximately four percentage units but was supplemented with crystalline lysine, methionine, threonine, and tryptophan. The low CP diets provided 2.64, 2.58, 2.50, 2.42, or 2.36 Mcal/kg in Exp. 1 as well as 2.45, 2.40, 2.35, 2.30, or 2.25 Mcai/kg in Exp.2. In Exp. 1, a linear （P=0.03） increase in weight gain was observed with decreasing NE level while the ratio of gain to feed was unaffect- ed （P 〉 0.05） by NE level. There was a linear in-crease （P = 0.01 ） in the percentage of fat-free lean and a linear decrease （ P = 0.03 ） in the percentage of total fat with decreasing NE levels. In Exp. 2, a sig- nificant quadratic （ P = 0.03 ） effect of NE level was observed for weight gain. The ratio of gain to feed demonstrated a significant （P 〈0.01 ） quadratic effect with pigs fed 2.35 and 2.40 Mcal/kg of NE, Pigs fed the diet containing 2.40 Mcal/kg of NE had the lowest percentage of total fat （36. 95% ） and the highest percentage of fat-free lean （49.36%）. The overall results of these experiments indicate that feed- ing either a surplus or a deficiency of NE is detrimen- tal to both pig performance and carcass composition when low CP diets supplemented with CAA are fed. Our results indicate that when the CP content of the diet is reduced by four percentage units and the diet is properly supplemented with CAA, maximum per- formance and carcass quality will be obtained if the diet provides approximately 2.42 Mcal/kg of NE.

Effects of different dietary energy and protein levels and sex on growth performance,carcass characteristics and meat quality of F1 Angus x Chinese Xiangxi yellow cattle

Background：The experiment evaluated the effect of nutrition levels and sex on the growth performance,carcass characteristics and meat quality of F1 Angus × Chinese Xiangxi yellow cattle.Methods：During the background period of 184 d,23 steers and 24 heifers were fed the same ration,then put into a2×2×2 factorial arrangement under two levels of- dietary energy（TON：70/80%DM）,protein（CP：11.9/14.3%DM）and sex（S：male/female） during the finishing phase of 146 d.The treatments were-（1） high energy/low protein（HELP）,（2） high energy/high protein（HEHP）,（3） low energy/low protein（LELP） and（4） low energy/high protein（LEHP）.Each treatment used 6 steers and 6 heifers,except for HELP- 5 steers and 6 heifers.Results：Growth rate and final carcass weight were unaffected by dietary energy and protein levels or by sex.Compared with the LE diet group,the HE group had significantly lower dry matter intake（DMI,6.76 vs.7.48 kg DM/d）,greater chest girth increments（46.1 vs.36.8 cm）,higher carcass fat（19.9 vs.16.3%） and intramuscular fat content（29.9 vs.22.8%DM）.The HE group also had improved yields of top and medium top grade commercial meat cuts（39.9 vs.36.5%）.The dressing percentage was higher for the HP group than the LP group（53.4 vs.54.9%）.Steers had a greater length increment（9.0 vs.8.3 cm）,but lower carcass fat content（16.8 vs.19.4%） than heifers.The meat quality traits（shear force value,drip loss,cooking loss and water holding capacity） were not affected by treatments or sex,averaging 3.14 kg,2.5,31.5 and 52.9%,respectively.The nutritive profiles（both fatty and amino acid composition） were not influenced by the energy or protein levels or by sex.Conclusions：The dietary energy and protein levels and sex significantly influenced the carcass characteristics and chemical composition of meat but not thegrowth performance,meat quality traits and nutritive profiles.

Reduced protein diet with near ideal amino acid profile improves energy efficiency and mitigate heat production associated with lactation in sows

Background:The study objective was to test the hypothesis that 1)lowering dietary crude protein(CP)increases dietary energetic efficiency and reduces metabolic heat associated with lactation,and 2)excessive dietary leucine(Leu)supplementation in a low CP diet decreases dietary energetic efficiency and increases metabolic heat associated with lactation.Methods:Fifty-four lactating multiparous Yorkshire sows were allotted to 1 of 3 isocaloric diets(10.80 MJ/kg net energy):1)control(CON;18.75%CP),2)reduced CP with a near ideal or optimal AA profile(OPT;13.75%CP)and 3)diet OPT with excessive Leu(OPTLEU;14.25%CP).Sow body weight and backfat were recorded on day 1 and 21 of lactation and piglets were weighed on day 1,4,8,14,18,and 21 of lactation.Energy balance was measured on sows during early(day 4 to 8)and peak(day 14 to18)lactation,and milk was sampled on day 8 and 18.Results:Over 21-day lactation,sows fed OPT lost body weight and body lipid(P<0.05).In peak lactation,sows fed OPT had higher milk energy output(P<0.05)than CON.Sows fed OPTLEU tended(P=0.07)to have less milk energy output than OPT and did not differ from CON.Maternal energy retention was lower(P<0.05)in OPT and OPTLEU compared to CON sows,and did not differ between OPTLEU and OPT sows.Sows fed OPT had higher(P<0.05)apparent energy efficiency for milk production compared to CON.Heat production associated with lactation was lower(P<0.05)or tended to be lower(P=0.082),respectively,in OPT and OPTLEU compared to CON sows.Conclusion:The OPT diet,in peak lactation,improved dietary energy utilization for lactation due to less urinary energy and metabolic heat loss,and triggered dietary energy deposition into milk at the expense of maternal lipid mobilization.Leucine supplementation above requirement may reduce dietary energy utilization for lactation by decreasing the energy partitioning towards milk,partially explaining the effectiveness of OPT diet over CON diets.

Effects of Different Dietary Energy and Protein Levels on Meat Performance and Meat Quality of Jinghai Yellow Chickens

To investigate the effect of different dietary energy and protein levels on meat performance and meat quality of Jinghai yellow chickens, 480 43-day old Jinghai yellow chickens with similar weight were randomly divided into four experimental groups： experimental group 1 （protein 15%, metabolic energy 9.95 MJ/kg）, experimental group 2 （protein 16%, metabolic energy 10.95 MJ/kg）, experimental group 3 （protein 17%, metabolic energy 12.65 MJ/kg） and experimental group 4 （ protein 18%, metabolic energy 13.95 MJ/kg）, respectively. All chickens were slaughtered at 112-day old. The breast and leg muscles of Jinghai yellow chickens were collected, to determine the slaughter performance, conventional meat quality and muscle chemical indicators. The results indicated that dressing-out percentage and eviscerated yield percentage in four experimental groups were above 87.27% and 67.00%, respectively; other slaughter performance indicators exhibited no significant differences among various groups （P 〉 0.05 ） ; breast muscle color of hens in experimental group 4 varied significantly from that in other three groups （ P 〈 0.05 ） ; leg muscle color of hens in experimental group 2 varied extremely significantly from that in other three groups （ P 〈 0.01 ） ; water-holding capacity of breast muscles of hens in experimental group 3 was significantly higher than that in experimental group 4 （P 〈 0.05 ） ; thiamine content of breast muscles of cocks in experimental group 3 was significandy higher than that in experimental group 2 （ P 〈 0.05 ） ; however, other properties exhibited no significant differenees among various groups （P 〉 0.05 ）.

Energy and Protein Requirements for 0-to 4-Week-Age Gushi Chicks

In this experiment, 540 male and female chicks of 1-day-age were selected respectively and reared separately. 3×3 factorial experiment was designed. Gushi chicks were fed with nine diets under different metabolizable energy (ME) and crude protein (CP). Effects of different diets on the performances and carcass ingredients of 0-to 4-week-age Gushi chicks were studied by rearing, comparative slaughter and metabolism experiments. Based on the results of three experiments, ME intake per kilogram metabolic weight was regarded as dependent variable (Y) ,and its corresponding net energy (NE) deposition per kilogram metabolic weight was regarded as independent variable (X) , respectively. Lineal regressive analysis was made according to the mathematic model: Y=a+bX. Maintenance ME( MEm) and the converted coefficient from net energy for gain( NEg) to ME for gain( MEg) were found out, then ME requirement was divided into MEm+ MEg, which is ME=359.14 W0.75+ 10.47△W; CP intake and corresponding metabolic weight were regarded as dependent variable and independent variable respectively, and then regressive analysis was made. The result was CP=0. 57 + 8. 21 W0.7S. Consequently, average daily gain (ADG), average daily feed intake and metabolic weight were put into the regressive equations, and then ME and CP requirements for 0- to 4-week-age Gushi chicks were obtained, 12.38 MJ kg-1 and 20.13%, respectively, the ratio of CP to ME was 16. 26 g MJ-1.

Distribution of Vibrational Energy Levels of Protein Molecular Chains

The distributions of the quantum vibrational energy levels of the protein molecular chain are found by the discretely nonlinear Schr?dinger equation appropriate to protein obtained from the Davydov theory. The results calculated by this method are basically consistent with the experimental values. Furthermore, the energy spectra at high excited states have also been obtained for the molecular chain which is helpful in researching the properties of infrared absorption and Raman scattering of the protein molecules.

Equilibrium folding and unfolding dynamics to reveal detailed free energy landscape of src SH3 protein by magnetic tweezers

Src SH3 protein domain is a typical two-state protein which has been confirmed by research of denaturant-induced unfolding dynamics.Force spectroscopy experiments by optical tweezers and atomic force microscopy have measured the force-dependent unfolding rates with different kinds of pulling geometry.However,the equilibrium folding and unfolding dynamics at constant forces has not been reported.Here,using stable magnetic tweezers,we performed equilibrium folding and unfolding dynamic measurement and force-jump measurement of src SH3 domain with tethering points at its N-and C-termini.From the obtained force-dependent transition rates,a detailed two-state free energy landscape of src SH3 protein is constructed with quantitative information of folding free energy,transition state barrier height and position,which exemplifies the capability of magnetic tweezers to study protein folding and unfolding dynamics.

Dietary lipid and gross energy affect protein utilization in the rare minnow Gobiocypris rarus

An 8-week feeding trial was conducted to detect the optimal dietary protein and energy, as well as the ef fects of protein to energy ratio on growth, for the rare minnow( Gobiocypris rarus), which are critical to nutrition standardization for model fi sh. Twenty-four diets were formulated to contain three gross energy(10, 12.5, 15 kJ/g), four protein(20%, 25%, 30%, 35%), and two lipid levels(3%, 6%). The results showed that optimal dietary E/P was 41.7–50 kJ/g for maximum growth in juvenile rare minnows at 6% dietary crude lipid. At 3% dietary lipid, specifi c growth rate(SGR) increased markedly when E/P decreased from 62.5 kJ/g to 35.7 kJ/g and gross energy was 12.5 kJ/g, and from 75 kJ/g to 42.9 kJ/g when gross energy was 15.0 kJ/g. The optimal gross energy was estimated at 12.5 kJ /g and excess energy decreased food intake and growth. Dietary lipid exhibited an apparent protein-sparing eff ect. Optimal protein decreased from 35% to 25%–30% with an increase in dietary lipid from 3% to 6% without adversely ef fecting growth. Dietary lipid level af fects the optimal dietary E/P ratio. In conclusion, recommended dietary protein and energy for rare minnow are 20%–35% and 10–12.5 k J/g, respectively.

Metabolomic changes in fatty liver can be modified by dietary protein and calcium during energy restriction

AIM： To characterise the effect of energy restriction （ER） on liver lipid and primary metabolite profile by using metabolomic approach. We also investigated whether the effect of energy restriction can be further enhanced by modification of dietary protein source and calcium. METHODS： Liver metabolomic profile of lean and obese C57BI/6J mice （n = 10/group） were compared with two groups of weight-reduced mice. ER was performed on control diet and whey protein-based high-calcium diet （whey ＋ Ca）. The metabolomic analyses were performed using the UPLC/MS based lipidomic platform and the HPLC/MS/MS based primary metabolite platform.RESULTS： ER on both diets significantly reduced hepatic lipid accumulation and lipid droplet size, while only whey ＋ Ca diet significantly decreased blood glucose （P 〈 0.001） and serum insulin （P 〈 0.01）. In hepatic lipid species the biggest reduction was in the level of triacylglycerols and cerarnides while the level of cholesterol esters was significantly increased during ER. Interestingly, diacylglycerol to phospholipid ratio, an indicator of relative amount of diabetogenic diglyceride species, was increased in the control ER group, but decreased in the whey ＋ Ca ER group （P 〈 0.001, vs obese）. ER on whey ＋ Ca diet also totally reversed the obesity induced increase in the relative level of lipotoxic cerarnides （P 〈 0.001, vs obese; P 〉 0.05, vs lean）. These changes were accompanied with up-regulated TCA cycle and pentose phosphate pathway rnetabolites. CONCLUSION： ER-induced changes on hepatic rnetabolornic profile can be significantly affected by dietary protein source. The therapeutic potential of whey protein and calcium should be further studied.

Screening and identification of bioactive compounds from citrus against non-structural protein 3 protease of hepatitis C virus genotype 3a by fluorescence resonance energy transfer assay and mass spectrometry

BACKGROUND Hepatitis C virus genotype 3a(HCV G3a)is highly prevalent in Pakistan.Due to the elevated cost of available Food and Drug Administration-approved drugs against HCV,medicinal natural products of potent antiviral activity should be screened for the cost-effective treatment of the disease.Furthermore,from natural products,active compounds against vital HCV proteins like non-structural protein 3(NS3)protease could be identified to prevent viral proliferation in the host.AIM To develop cost-effective HCV genotype 3a NS3 protease inhibitors from citrus fruit extracts.METHODS Full-length NS3 without co-factor non-structural protein 4A(NS4A)and codon optimized NS3 protease in fusion with NS4A were expressed in Escherichia coli.The expressed protein was purified by metal ion affinity chromatography and gel filtration.Citrus fruit extracts were screened using fluorescence resonance energy transfer(FRET)assay against the protease and polyphenols were identified as potential inhibitors using electrospray ionization-mass spectrometry(MS)/MS technique.Among different polyphenols,highly potent compounds were screened using molecular modeling approaches and consequently the most active compound was further evaluated against HCV NS4A-NS3 protease domain using FRET assay.RESULTS NS4A fused with NS3 protease domain gene was overexpressed and the purified protein yield was high in comparison to the lower yield of the full-length NS3 protein.Furthermore,in enzyme kinetic studies,NS4A fused with NS3 protease proved to be functionally active compared to full-length NS3.So it was concluded that co-factor NS4A fusion is essential for the purification of functionally active protease.FRET assay was developed and validated by the half maximal inhibitory concentration(IC50)values of commercially available inhibitors.Screening of citrus fruit extracts against the native purified fused NS4A-NS3 protease domain showed that the grapefruit mesocarp extract exhibits the highest percentage inhibition 91%of protease activity.Among the compounds identified by LCMS analysis,hesperidin showed strong binding affinity with the protease catalytic triad having S-score value of-10.98.CONCLUSION Fused NS4A-NS3 protease is functionally more active,which is effectively inhibited by hesperidin from the grapefruit mesocarp extract with an IC50 value of 23.32μmol/L.

Association of Protein Energy Wasting with Income in CKD Stage 3 Patients

Protein energy wasting (PEW) is a major challenge in CKD. Objective: To assess PEW in predialysis patients on their first visit to a nephrologist. Methods: Three day dietary intake of 484 CKD stage 3 patients was taken. Appetite was assessed with ADAT. Patients were divided into groups based on appetite and BMI. Results: Male and female parameters are serum albumin 3.7 ± 0.84/3.68.8 ± 0.81 g/dL, total protein 7.02 ± 1.27/6.94 ± 1.26 g/dL, creatinine 4.68 ± 4.19/3.74 ± 3.36 mg% creatinine clearance 33.22 ± 30.48/37.55 ± 33.87 ml/minute, BMI 22.60 ± 4.29/23.43 ± 4.77kg/m2 energy/kg 16.97 ± 0.65/16.8 ± 0.64, protein g/kg 0.65 ± 0.28/0.64 ± 0.30, carbohydrate g/kg 2.98 ± 1.54/2.98 ± 0.1.36, fat g/kg 2.98 ± 0.23/2.79 ± 0.22, respectively. As appetite decreased, dietary protein and energy intake decreased significantly. Appetite in males and females: Average 14.46%, 4.13%, poor 9.7%, 18.18%, anorexic 13.2%, 7.4%. Income had strong correlation with BMI (p 0.000), dietary protein (p 0.000), energy (p 0.000) and carbohydrate (p 0.000). Appetite correlated with creatinine (p 0.019), dietary energy, protein, carbohydrate and fat (p 0.000) intake. BMI correlated (p 0.000) with fat, carbohydrate, energy and creatinine clearance. ANOVA showed significant difference within and between appetite groups in energy, protein, fat, carbohydrate, creatinine clearance (p 0.000) and serum albumin (p 0.025). There was significant difference in protein (p 0.026), energy intake (p 0.000) and creatinine clearance (p 0.038) within and between BMI groups. Based on income, there was significant difference among groups in BMI (p 0.000), energy (p 0.019), protein (p 0.031) and albumin (0.001).