Effect of milk protein intake and casein-to-whey ratio in breakfast meals on postprandial glucose,satiety ratings,and subsequent meal intake

Whey and casein proteins differentially affect postprandial blood glucose and satiety mechanisms, with relevance for type 2 diabetes and obesity. Therefore, the purpose of this work was to investigate the effect of the casein-to-whey protein ratio and total protein concentration of milks consumed with cereal on postprandial blood glucose, appetite ratings, and subsequent food intake in a randomized, controlled, double-blinded study with healthy young adults (n = 32, 23.4 ± 3.1 yr, body mass index = 22.2 ± 2.5 kg/m²). Fasted participants consumed milk (250 mL) with either 80:20 or 40:60 (modified) casein-to-whey protein ratios at commercially normal (3.1%, wt) or high protein (9.3%, wt) concentration, or control (water with whey permeate), each along with 2 servings of oat-based breakfast cereal. Blood glucose concentrations were determined from finger prick blood samples and appetite was assessed using visual analog scales. Participants consumed a measured ad libitum pizza lunch at 120 min and blood glucose determination and appetite assessment continued following the lunch meal (140–200 min) to observe the second meal effect. Pre-lunch (0–120 min) incremental area under the curve (iAUC) and mean change from baseline blood glucose were reduced with consumption of all milk treatments relative to control. However, we found no differences between all treatments on pre-lunch appetite change from baseline and total area under the curve (tAUC) or lunch meal food intake. In terms of protein concentration results, high protein (9.3%, wt) treatments contrasted to normal protein (3.1%, wt) treatments lowered blood glucose change from baseline and iAUC, and post-lunch appetite change from baseline and tAUC. Protein ratio showed a modest effect in that modified (40:60) protein ratio lowered pre-lunch blood glucose change from baseline but not iAUC, and normal (80:20) protein ratio lowered pre-lunch appetite change from baseline but not tAUC. Therefore, high-carbohydrate breakfast meals with increased protein concentration (9.3%, wt) could be a dietary strategy for the attenuation of blood glucose and improved satiety ratings after the second meal.     

脂肪酸通过神经肽和胃肠激素调节食欲的机制

脂肪酸是参与食欲调节和能量稳态的重要因子,包括改善机体组成、葡萄糖稳态、血脂谱和体重平衡。脂肪酸促食欲效应由中枢神经系统及胃肠激素联合介导。一方面脂肪酸通过影响食欲调控中枢（下丘脑）及其它中枢神经系统区域（包括中脑边缘多巴胺系统、纹状体、杏仁体等）构成的复杂神经网络参与调控食欲、摄食行为及能量稳态;另一方面,脂肪酸通过刺激内分泌激素（如瘦素、胰岛素、胃饥饿素）分泌将胃肠道信号上传至中枢神经系统,组成脑-肠肽轴调节食欲及摄食行为。不同链长脂肪酸在机体发挥的作用不同,这些效应结合在一起,对机体食欲产生总体影响。本文综述脂肪酸通过中枢和外周对食欲的调节机制,重点阐述不同链长脂肪酸的多种食欲调节作用和信号传导路径。     

Use of individual cow milk recording data at the start of lactation to predict the calving to conception interval

Milk recording data collected in 2,128 dairy herds in England and Wales between 2004 and 2006 were used to predict the calving to conception intervals. The average cumulative milk production was 8,200 kg. Conception (or not) within 5 intervals measured in days (20 to 60 d, 61 to 81 d, 82 to 102 d, 103 to 123 d, 124 to 144 d) was modeled as a function of milk yields and milk constituents at the start of lactation using multilevel discrete-time survival models. Milk yield, weight and percentage of fat, protein, and lactose, and somatic cell counts on the first 2 test-days of lactation were corrected for either stage of lactation alone or stage of lactation and time of year. Five hundred and 1,628 herds, respectively, were used for parameter estimation and cross-validation. Covariates were retained in the final model if their coefficient was at least twice its standard error and their inclusion resulted in a decrease in the deviance. Overall, 73% of cows recalved. The percentage of cows that had conceived by d 20, 61, 82, 103, 124, and 145 were 0.5, 7.3, 17.9, 29.3, 38.7, and 46, respectively. The probability of conception before 145 d in milk increased with lower milk production on the second test-day, higher percentage of protein on the second test-day, and higher percentage of lactose on the first test-day. Positive associations were of a limited magnitude but nonetheless significant with the percentage of protein on the first test-day, the percentage of butterfat on the first test-day, and somatic cell count on both test-days. The model predicted the probability of conception in the cross validation data set very well. Despite the common use of fat to protein ratio as a measure of energy balance, this parameter exhibited wide variation with stage of lactation and time of the year and had a much-reduced ability to predict an early conception compared with other combinations of milk quantity and constituents.     

Influence of proteolysis of milk on the whey protein to total protein ratio as determined by capillary electrophoresis

Capillary electrophoresis (CE) was used to determine the whey protein to total protein ratio in raw and UHT milk samples with different degrees of proteolysis caused by storage. In raw milks, the analysis of samples taken at regular times demonstrated the influence of proteolysis in the whey protein to total protein determination, which was overestimated after 4 d of storage. In UHT milks, the overestimation of the whey protein to total protein ratio took place after 30 or 60 d of storage. However, the ratios alphaS1-CN/beta-CN and alphas1-CN/kappa-CN permitted detection of the samples of raw or UHT milk with degraded proteins. The distorted capillary electrophoretic pattern obtained for UHT milks made necessary an integration of the electropherograms in a "valley-to-valley" way. Results for raw milk samples were identical when "valley-to-valley" was compared to standard integration techniques. This CE method could be considered an alternative method to derivative spectroscopy for the determination of the whey protein to total protein of milk and could be used to detect samples with proteolysis.     

乳及乳制品中稳定碳同位素比的研究

目的建立元素分析-稳定同位素比质谱法(elemental analyzer-isotope ratio mass spectrometry,EA-IRMS)测定乳及乳制品中δ~(13)C值的检测方法。方法本文研究了样品前处理、进样量、不同标样标定等对测定结果的影响,通过优化仪器工作条件,建立了稳定同位素比质谱仪测定乳及乳制品中碳同位素比(δ~(13)C)的方法,并用该方法对20余种品牌的乳及乳制品及3位哺乳期的母乳进行检测。结果研究数据表明,不同种类的乳汁δ~(13)C数值有明显差异:牛乳及其制品δ~(13)C值位于-16.50‰~-20.42‰,羊乳及其制品δ~(13)C值位于-22.38‰~-22.95‰,母乳δ~(13)C值为-23.17‰~-27.30‰。结论本方法根据羊乳和与牛乳δ~(13)C特征数值的差异,可以进行乳品纯正性判别。该方法精密度为0.04‰~0.24‰,小于常规0.3‰的要求,稳定性好、准确度高,操作简便,为乳及乳制品的鉴别提供了技术支持和理论基础。     

Varying the ratio of Lys:Met while maintaining the ratios of Thr:Phe,Lys:Thr,Lys:His,and Lys:Val alters mammary cellular metabolites,mammalian target of rapamycin signaling,and gene transcription

Amino acids are not only precursors for but also signaling molecules regulating protein synthesis. Regulation of protein synthesis via AA occurs at least in part by alterations in the phosphorylation status of mammalian target of rapamycin (mTOR) pathway proteins. Although the ideal profile of Lys:Met to promote milk protein synthesis during established lactation in dairy cows has been proposed to be 3:1, aside from being the most-limiting AA for milk protein synthesis, the role of Met in other key biologic pathways such as methylation is not well characterized in the bovine. The objective of this study was to determine the influence of increasing supplemental Met, based on the ideal 3:1 ratio of Lys to Met, on intracellular metabolism related to protein synthesis and mTOR pathway phosphorylation status. MAC-T cells, an immortalized bovine mammary epithelial cell line, were incubated (n = 5 replicates/treatment) for 12 h with 3 incremental doses of Met while holding Lys concentration constant to achieve the following: Lys:Met 2.9:1 (ideal AA ratio; IPAA), Lys:Met 2.5:1 (LM2.5), and Lys:Met 2.0:1 (LM2.0). The ratios of Thr:Phe (1.05:1), Lys:Thr (1.8:1), Lys:His (2.38:1), and Lys:Val (1.23:1) were the same across the 3 treatments. Applying gas chromatography–mass spectrometry metabolomics revealed distinct clusters of differentially concentrated metabolites in response to Lys:Met. Lower Phe, branched-chain AA, and putrescine concentrations were observed with LM2.5 compared with IPAA. Apart from greater intracellular Met concentrations, further elevations in Met level (LM2.0) led to greater intracellular concentrations of nonessential AA (Pro, Glu, Gln, and Gly) compared with IPAA and greater essential AA (EAA; Met, Ile, and Leu) and nonessential AA (Pro, Gly, Ala, Gln, and Glu) compared with LM2.5. However, compared with IPAA, mRNA expression of β-casein and AA transporters (SLC7A5, SLC36A1, SLC38A2, SLC38A9, and SLC43A1) and mTOR phosphorylation were lower in response to LM2.5 and LM2.0. Overall, the results of this study provide evidence that increasing Met while Lys and the ratios of Phe, Thr, His, and Val relative to Lys were held constant could increase the concentration and utilization of intracellular EAA, in particular branched-chain AA, potentially through improving the activity of AA transporters partly controlled by mTOR signaling. Because EAA likely are metabolized by other tissues upon absorption, a question for future in vivo studies is whether formulating diets for optimal ratios of EAA in the metabolizable protein is sufficient to provide the desired levels of these AA to the mammary cells.     

Effect of kappa-casein and beta-lactoglobulin loci on milk production traits and reproductive performance of Holstein cows

The effects of kappa-casein (kappa-CN) and beta-lactoglobulin (beta-LG) loci on milk production traits (milk, fat, protein, and lactose yield, fat, protein, and lactose content) and reproductive performance (gestation length, calving interval, age at first and second calving, number of services per conception) was estimated for 278 Holstein cows in the first 2 lactations. Genotypes of kappa-CN and beta-LG were determined by alkaline and acidic polyacrylamide gel electrophoresis. Milk production was recorded daily. Single-trait, mixed, linear models were used for the statistical analysis of the data. Results indicated that kappa-CN genotypes affected significantly protein yield and content (genotype AB > genotype AA). A tendency for increased milk and fat yield of animals having AB kappa-CN genotype was also found. Fat content and lactose yield and content were not affected. In the beta-LG system, significant differences were detected for milk yield (AB > AA), fat yield (BB and AB > AA), fat content (BB > AA and AB), and lactose yield (AB > AA). A tendency for higher protein yield was also observed (AB > AA). The beta-LG locus had no significant effect on protein and lactose content. No associations between polymorphisms at the kappa-CN locus and reproductive performance were found. There was a tendency, however, for cows with AB genotype to have older age at first and second calving. In the beta-LG system, cows with AA genotype had significantly shorter gestation length than did those with AB or BB genotype. No differences were detected between beta-LG polymorphisms for the other reproductive traits.     

水牛乳蛋白质的组成

分析了摩拉水牛（M）、尼里-拉菲水牛（N）、一代杂交水牛（F1）、二代杂交水牛（F2）和高代杂交水牛（Fh）5个品代水牛的乳蛋白主要组分的相对百分比含量．同时分析了总氨基酸组成及钙、磷含量。结果表明，水牛乳蛋白的主要组分有：α-乳清蛋白（α-LA）、β-乳球蛋白（β-LG）、免疫球蛋白轻链（IgG—L）和重链（IgG—H）、αs1-酪蛋白（αs1-CN）、αs2-酪蛋白（αs2-CN），β-酪蛋白（β-CN）、κ-酪蛋白（κ—CN）、血清白蛋白（SA）和乳铁蛋白（LF）等；CN在水牛乳蛋白中占优势，与荷斯坦牛乳相比，水牛乳中CN的质量分数稍低，而且各品代水牛乳中的CN有显著性差异（P〈0．05）；乳清蛋白中β-LG含量最高；杂交水牛乳蛋白高于纯种摩拉水牛和尼里一拉菲水牛，差异显著（P〈0．05）：各品代水牛乳的氨基酸比例比较接近；不同品代水牛乳中钙、磷含量没有显著性差异。     

Solubility of commercial milk protein concentrates and milk protein isolates

High-protein milk protein concentrate (MPC) and milk protein isolate (MPI) powders may have lower solubility than low-protein MPC powders, but information is limited on MPC solubility. Our objectives in this study were to (1) characterize the solubility of commercially available powder types with differing protein contents such as MPC40, MPC80, and MPI obtained from various manufacturers (sources), and (2) determine if such differences could be associated with differences in mineral, protein composition, and conformational changes of the powders. To examine possible predictors of solubility as measured by percent suspension stability (%SS), mineral analysis, Fourier transform infrared (FTIR) spectroscopy, and quantitative protein analysis by HPLC was performed. After accounting for overall differences between powder types, %SS was found to be strongly associated with the calcium, magnesium, phosphorus, and sodium content of the powders. The FTIR score plots were in agreement with %SS results. A principal component analysis of FTIR spectra clustered the highly soluble MPC40 separately from the rest of samples. Furthermore, 2 highly soluble MPI samples were clustered separately from the rest of the MPC80 and MPI samples. We found that the 900 to 1,200 cm⁻¹ region exhibited the highest discriminating power, with dominant bands at 1,173 and 968 cm⁻¹, associated with phosphate vibrations. The 2 highly soluble MPI powders were observed to have lower κ-casein and α-_S1-casein contents and slightly higher whey protein contents than the other powders. The differences in the solubility of MPC and MPI were associated with a difference in mineral composition, which may be attributed to differences in processing conditions. Additional studies on the role of minerals composition on MPC80 solubility are warranted. Such a study would provide a greater understanding of factors associated with differences in solubility and can provide insight on methods to improve solubility of high-protein milk protein concentrates.     

不补料酶膜耦合反应制备牛乳蛋白ACE抑制肽的研究

为了克服传统酶解技术的不足,提高酶解反应效率,采用不补料酶膜耦合反应制备牛乳蛋白ACE抑制肽。考察超滤膜对牛乳蛋白及酶的截留、膜通量及滤出液的ACE抑制率和IC₅₀的影响,确定出超滤膜最佳的截留分子量为5000 u;研究了反应时间、底物浓度、加酶量和循环泵转速对蛋白转化率的影响,确定最优酶解工艺条件:底物浓度7%（W/W）、加酶量2000 U/g、反应时间100 min、循环泵转速100 r/min,该条件下蛋白转化率、单位酶产肽量、ACE抑制率分别达到53.16%、39.59 g肽/g酶、76.81%,与对照组相比蛋白转化率、单位酶产肽量、ACE抑制率分别提高了16.99%、16.61%、18.55%;采用高效凝胶过滤色谱法测定超滤液中ACE抑制肽的分子量分布,发现样品中分子量≤2253 u组分质量分数为94.62%,超滤膜对截留分子量大小控制准确。因此不补料酶膜耦合反应可为酶法制备牛乳蛋白ACE抑制肽提供一种更为高效的方法。      

牛乳重要营养品质特征的研究进展

牛乳营养组成非常丰富,乳蛋白和乳脂肪是表征牛奶营养品质的重要指标,并受诸多因素的影响。本文综述遗传、环境和饲养管理等因素对乳蛋白和乳脂肪组分及含量的影响,以及牛乳蛋白和乳脂肪酸的特征性含量和比值。     

Effect of milk protein standardization using different methods on the composition and yields of Cheddar cheese

Two sets of Cheddar cheese were made in which the milk protein level (%, wt/wt) was increased from 3.3 (Control A, CA) to 3.6 (set A) or from 3.3 (control B, CB) to 4.0 (set B) by the addition of phosphocasein (PC), milk protein concentrate (MPC), or freshly prepared ultrafiltered milk retentate (UFR). The cheeses were denoted CA, PCA, MPCA, and UFRA from set A, and CB, PCB, MPCB, and UFRB, from set B, respectively. The level of cheese moisture decreased significantly on increasing milk protein level from 3.3 to 3.6 or 4.0% (wt/wt), but was not affected significantly by the method of protein standardization. The percentage milk fat recovered to cheese increased significantly on increasing the level of milk protein from 3.3 to 3.6% (wt/wt) with PC, and from 3.3 to 4.0% (wt/wt) with PC, MPC, and UFR. Increasing milk protein level from 3.3 to 4.0% (wt/wt) with PC significantly increased the percentage of milk protein recovered to cheese. Actual cheese yield increased significantly with milk protein level. The yield of cheese per 100 kg of milk normalized to reference levels of fat (3.4%, wt/wt) and casein (2.53%, wt/wt) indicated no significant effects of protein content or standardization treatment on yield. However, the moisture-adjusted yield per 100 kg of milk with reference levels of fat and casein increased significantly on increasing the protein content from 3.3 to 3.6% (wt/wt) with MPC and from 3.3 to 4.0% (wt/wt) with PC, MPC, and UFR.     

The effects of heat stress on protein metabolism in lactating Holstein cows

Heat stress (HS) decreases milk protein synthesis beyond what would be expected based on the concomitant reduction in feed intake. The aim of the present study was to evaluate the direct effects of HS on milk protein production. Four multiparous, lactating Holstein cows (101 ± 10 d in milk, 574 ± 36 kg of body weight, 38 ± 2 kg of milk/d) were individually housed in environmental chambers and randomly allocated to 1 of 2 groups in a crossover design. The study was divided into 2 periods with 2 identical experimental phases (control phase and trial phase) within each period. During phase 1 or control phase (9 d), all cows were housed in thermal neutral conditions (TN; 20°C, 55% humidity) and fed ad libitum. During phase 2 or treatment phase (9 d), group 1 was exposed to cyclical HS conditions (32 to 36°C, 40% humidity) and fed ad libitum, whereas group 2 remained in TN conditions but was pair-fed (PFTN) to their HS counterparts to eliminate the confounding effects of dissimilar feed intake. After a 30-d washout period in TN conditions, the study was repeated (period 2), inverting the environmental treatments of the groups relative to period 1: group 2 was exposed to HS and group 1 to PFTN conditions. Compared with PFTN conditions, HS decreased milk yield (17.0%), milk protein (4.1%), milk protein yield (19%), 4% fat-corrected milk (23%), and fat yield (19%). Apparent digestibility of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, crude protein, and ether extract was increased (11.1–42.9%) in HS cows, as well as rumen liquor ammonia (before feeding 33.2%; after feeding 29.5%) and volatile fatty acid concentration (45.3%) before feeding. In addition, ruminal pH was reduced (9.5 and 6% before and after feeding, respectively) during HS. Heat stress decreased plasma free amino acids (AA; 17.1%) and tended to increase and increased blood, urine, and milk urea nitrogen (17.2, 243, and 24.5%, respectively). Further, HS cows had reduced plasma glucose (8%) and nonesterified fatty acid (39.8%) concentrations compared with PFTN controls. These data suggest that HS increases systemic AA utilization (e.g., decreased plasma AA and increased nitrogen excretion), a scenario that limits the AA supply to the mammary gland for milk protein synthesis. Furthermore, the increase in AA requirements during HS might represent the increased need for gluconeogenic precursors, as HS is thought to prioritize glucose utilization as a fuel at the expense of nonesterified fatty acids.