Maternal obesity exacerbates the responsiveness of offspring BALB/c mice to cow's milk protein-induced food allergy

Food allergy has become a significant public health problem affecting a large number of people worldwide.Maternal obesity causes inflammation and alters the immune system of offspring,which may exacerbate their food allergy.The aim of this study was to determine whether offspring mice born to obese mothers would have more serve reactions to cow's milk protein-induced food allergy,and further investigate the underlying mechanisms.Female offspring BALB/c mice of mothers with normal and high-fat diets were sensitized withβ-lactoglobulin(BLG),respectively.Maternal obesity increased the serum immunoglobulin E and mouse mast cell protease levels,though did not have significant influence on anaphylactic symptom score,core temperature and diarrhea rate of offspring mice after BLG sensitization.Furthermore,maternal obesity led to a lower level of occludin mRNA expression in BLG-sensitized mice.The mice born to obese mothers exhibited increased mRNA expression levels of GATA-3,interleukin(IL)-4 and IL-10 in jejunum after BLG sensitization,indicating maternal obesity intensified Th2-type biased immune responses.In conclusion,maternal obesity exerted exacerbating effects on the responsiveness of their offspring to cow's milk protein sensitization.

Use of fluorometry for determination of skim milk powder adulteration in fresh milk

A FAST (fluorescence of advanced Maillard products and Soluble Tryptophan) method for identification of recon- stituted milk made from skim milk powder in the fresh milk was developed. Considering milk and skim milk powders variations from different seasons and countries, milk was collected from different dairy farms in different seasons and skim milk powders were collected from different countries to measure the Tryptophan (Trp), advanced Maillard products (AMP) fluorescence values. The results showed that there were differences (P<0.01) between raw and reconstituted milk. The plot of values in each mixed level of raw and reconstituted milk had a correlation coefficient >0.97. The FAST method is a simple, rapid, low-cost and sensitive method enabling the detection of 5% reconstituted milk in fresh milk. The measurement of the Trp, AMP fluorescence values and calculation of the FAST index is a suitable method for large-scale monitoring of fresh milk samples.

Development and Optimization of an Alternative Methodology for Detection of Milk Adulteration by Water

This paper presents a process which allows the determination of milk adulteration by water. Considering the limitations of the reference method （cryoscopy）, the proposed methodology matches electrical conductivity and cryoscopy index measurements. Based on statistical procedures, a linear descriptive model is achieved to quantify adulteration of milk with water. It is also shown that electrical conductivity measurements allow the detection of adulteration with water and sodium chloride.

Study of Calcium and Sodium Behavior to Identify Milk Adulteration Using Flame Atomic Absorption Spectrometry

A fast and direct method for determination of milk adulteration by monitoring of calcium and sodium concentrations variations was described. Milk samples were furnished by a dairy company located at S?o Carlos (S?o Paulo State, Brazil) and and spiked with tap-water, whey, hydrogen peroxide, synthetic urine, urea and synthetic milk in the ranged from 5% to 50% (v/v), expect for caustic soda. Caustic soda was added in the milk until establish the original pH. The milk samples were analyzed by using flame atomic absorption spectrometry (FAAS) and no acid digestion process was required. Results showed a significant decrease in the Na and Ca concentrations with addition of synthetic milk and tap-water, a nonlinear variation with addition of synthetic urine, whey and hydrogen peroxide and a largest increase in the Na concentration with addition of NaOH. Correlation between Na and Ca concentrations in pure and adulterated milk were evaluated by paired t-test at a 95% confidence level. Results showed that the method proposed is efficient to identify samples adulterated with tap-water, caustic soda, synthetics milk and urine.

Validation of the Methods for Detection the Non-Milk Fat in a Mixture of Milk Fat and Palm Oil

Milk fat contains a variety of nutritive and health-promoting compounds that guard against some disease. In the current system of global competition, when the quality of milk and milk products is not an option but rather a requirement, therefore, determining the purity of milk fat is critical. This study aims to validate analytical methods for detecting palm oil in a mixture of milk fat and palm oil. Methods of this study was involved detection of non-milk fat in fat blinders by determining the saponification value, iodine number, refractive index, butyro refractometer reading, Gas chromatography, Reverse Phase High-performance liquid chromatography, and Fourier transforms Infrared. The results of this study revealed that the saponification value, Iodine number, refractive index, and Butyro Reading could be used to detect the addition of palm oil by a level of 10% - 20% or more to the milk. The level of some fatty acids in the milk as determined by GC, such as myristic acid (C14:0), palmitic acid (C16:0), and stearic acid (C18:0), is correlated well with the level of adding palm oil to milk fat. The determination of cholesterol and β-sito-sterol content by RP-HPLC could be used for the detection of the addition of palm oil to milk fat. The spectrum behavior produced by FTIR spectroscopy in this adulterated sample is almost the same, so this technique could not be used to detect the palm oil in milk fat.

Very early onset perinatal constipation:Can it be cow’s milk protein allergy?

Delayed passage of meconium or constipation during the perinatal period is traditionally regarded as a signal to initiate further work up to evaluate for serious diagnoses such as Hirschsprung’s disease(HD),meconium ileus due to Cystic Fibrosis,etc.The diagnosis of HD particularly warrants invasive testing to confirm the diagnosis,such as anorectal manometry or rectal suction biopsy.What if there was another etiology of perinatal constipation,that is far lesser known?Cow’s milk protein allergy(CMPA)is often diagnosed in infants within the first few weeks of life,however,there are studies that show that the CMPA allergen can be passed from mother to an infant in-utero,therefore allowing symptoms to show as early as day one of life.The presentation is more atypical,with perinatal constipation rather than with bloody stools,diarrhea,and vomiting.The diagnosis and management would be avoidance of cow's milk protein within the diet,with results and symptom improvement in patients immediately.Therefore,we discuss whether an alternative pathway to address perinatal constipation should be further discussed and implemented to potentially avoid invasive techniques in patients.This entails first ruling out CMPA with safe,noninvasive techniques with diet modification,and if unsuccessful,then moving forward with further diagnostic modalities.

婴幼儿奶粉中多种掺假物近红外高光谱图像检测方法

奶粉市场是食品掺假行为频发领域,其中婴幼儿配方奶粉价格高,其质量是消费者、生产企业和执法部门关注的重点。近红外高光谱成像(Near infrared-hyperspectral imaging,NIR-HSI)技术结合化学计量学和机器学习算法可以检测奶粉中单一掺假物含量。基于NIR-HSI技术研究了不同品牌婴幼儿奶粉中多掺假物(三聚氰胺、香兰素和淀粉)的定量预测。对基于像素点预处理后的高光谱图像划分感兴趣区域(Region of interest,ROI),提取ROI平均光谱。基于经典的过滤式特征选择算法拉普拉斯分数(Laplacian score)(无监督)和ReliefF(有监督)挑选建模关键变量,建立偏最小二乘回归模型(Partial least squares,PLS)。开发包含自定义选择层的一维卷积神经网络模型(One-dimensional convolutional neural networks,1DCNN)。自定义层根据权重系数绝对值,可确定重要波长变量。Laplacian score-PLS模型对预测集中奶粉、三聚氰胺、香兰素和淀粉质量分数预测结果均方根误差分别为0.1110%、0.0570%、0.0349%和0.3481%。ReliefF-PLS模型对预测集中奶粉、三聚氰胺、香兰素和淀粉预测结果均方根误差分别为0.1998%、0.0540%、0.0455%和0.1823%。1DCNN模型对预测集中奶粉、三聚氰胺、香兰素和淀粉质量分数预测结果均方根误差分别为0.8561%、0.0911%、0.0644%和0.2942%。对Laplacian score、ReliefF和自定义选择层挑选出的前15个重要波长进行对比分析,不同特征选择方法挑选的特征波长子集有所区别,但都选择1210、1474、1524、1680 nm等附近波长。基于ReliefF-PLS模型的可视化结果表明了其良好的预测能力。

基于近红外光谱技术的牦牛奶粉掺假检测与产地识别研究

牦牛奶粉的掺假检测和产地识别有助于保障食品安全、维护消费者权益,是促进乳制品市场健康发展的重要举措。传统的DNA检测方法和稳定同位素分析技术的检测周期长,难以满足快速、低成本现场分析的需求。针对以上问题,本研究建立了一种基于近红外光谱技术(Near-infrared Spectroscopy,NIRS)快速辨别牦牛奶粉掺假及产地的方法。收集了来自四川、甘肃、云南及青海的9个品牌的牦牛奶粉。在制备掺假样品之前,采用聚合酶链式反应(Polymerase Chain Reaction,PCR)技术和DNA凝胶电泳验证所收集的牦牛奶粉中是否掺杂了牛奶粉。完成验证后,进行掺假样品的制备以及近红外光谱数据的采集。采用K最邻近法(K-Nearest Neighbors,KNN)建立分类模型,偏最小二乘回归(Partial Least Squares Regression,PLSR)建立定量预测模型。通过优化光谱预处理方法和变量筛选方法进一步提升定量预测模型的预测能力。结果表明,KNN对牦牛奶粉掺假检测(纯牛奶粉、纯牦牛奶粉、掺杂着牛奶粉的牦牛奶粉)及产地识别(四川、甘肃、云南、青海)实现了100%的正确分类。掺假定量预测模型的校正集相关系数(R_(c))为0.9975,预测集相关系数(R_(p))为0.9913,预测集均方根误差(Root Mean Square Error of Prediction,RMSEP)为1.9823%,性能偏差比(Ratio of Performance to Deviation,RPD)为7.2522。本方法可快速、准确地预测牦牛奶粉中牛奶粉的掺杂以及牦牛奶粉产地的辨别,为牦牛奶粉的质量控制提供技术支持。

基于Real-time PCR法检测乳粉中牛源性成分定量研究

基于Real-timePCR建立了乳粉中牛源性成分相对定量检测方法,并对牛的特异性引物与探针进行了特异性、灵敏度和稳定性测试。通过模拟不同浓度牛乳粉与马乳粉混合样本,根据其△Ct值的函数关系进行线性拟合进而绘制标准曲线,建立乳粉中牛源性成分的相对定量检测。结果显示,该方法的最低检测限为0.00001 mg/mL,回收率为91.11%~119.2%,组间变异系数≤0.58%、组内变异系数≤1.44%。说明该方法在特异性与稳定性上适用于乳粉中牛源性成分及含量的掺假检测。

超高效液相色谱-串联质谱法鉴定驼乳及其制品中的动物源性成分

采用蛋白质组学技术建立了驼乳及其制品中的动物乳源性成分特异性肽生物标志物的鉴定方法。样品经脱脂、蛋白质提取、胰蛋白酶水解后,利用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱仪(UHPLC-Q/Exactive-HRMS)和Protein Pilot软件,实现了多肽生物标记物的鉴定;然后通过基本局部比对搜索工具(BLAST)与Uniprot数据库对比分析,筛选出了骆驼、家牛、水牛、牦牛、山羊、绵羊、驴和马共8个物种的22条肽生物标志物;最后利用超高效液相色谱-三重四极杆质谱(UHPLC-QqQ-MS)系统对这22条特征性多肽进行验证,采用多反应监测(MRM)模式建立了定量方法。实验优化了骆驼奶中酪蛋白的预处理方法,如冷冻脱脂、沉淀蛋白试剂和复溶液的选择等,并建立了基于生物标志物肽测定牛和山羊奶/奶粉掺假骆驼奶/奶粉的无标记定量方法。将牛、山羊和骆驼奶/奶粉等比例混合,分别对牛和山羊的特征肽段进行检测,结果显示,该方法对掺假液体乳/固体奶粉样品显示出良好的线性关系,抗干扰能力强,灵敏度高,重复性好,液态奶和奶粉的掺杂结果均与理论值接近。另外,该方法还被应用于11种骆驼奶和奶粉中家牛、水牛、牦牛、山羊、绵羊、驴和马乳源性成分的鉴别,为骆驼乳及乳制品中多种动物乳源性成分的鉴别提供了有效的技术平台。

近红外光谱结合siPLS法用于深度水解蛋白奶粉掺伪的快速检测

目的:建立深度水解蛋白奶粉中掺伪普通蛋白粉的快速检测方法。方法:向深度水解蛋白奶粉掺伪一定比例的牛乳清蛋白粉和植物蛋白粉,共制备171个掺伪样品,并采集近红外光谱;对采集的样品光谱使用SPXY法按3∶1比例划分为校正集和预测集,应用联合区间偏最小二乘法(siPLS)建立掺伪检测模型,并比较不同预处理方法下的建模效果。结果:SG一阶导预处理下建立的siPLS模型效果最好,其组合区间光谱范围为[1135~1239.5,1660~1764.5,2080~2184.5 nm],校正集相关系数R^(2)为0.9948,RMSECV值为0.0101,预测集相关系数R^(2)为0.9945,RMSEP值为0.0110,RPD值为13.5。结论:通过siPLS法筛选光谱区间建模,可提高模型的稳定性和预测精度,本方法操作简便,可用于深度水解蛋白奶粉中的掺伪蛋白粉的快速无损检测。

基于牛磺酸 叶酸和维A为特征指标羊乳掺假的 分析研究

为了初步准确判断羊乳是否掺假,通过建立高效液相色谱法和气相色谱法对纯牛羊乳及其乳粉制品中牛磺酸、叶酸和维A含量进行测定,总结不同样品中3种生长因子的含量变化规律,并结合主成分分析(PCA)建立研究方法。该方法简便且准确性高,可应用于羊乳的初步真伪鉴别,为建立健全乳品质量检验技术体系,保障乳品市场的规范化运行提供了依据。

同步荧光光谱技术结合支持向量机对掺杂牛奶智能判别研究

牛奶因其丰富的营养成分和易消化吸收的特点,受到消费者的青睐。牛奶掺杂行为的产生使得牛奶制品质量备受关注,快速、便捷地鉴别乳品质量对于乳制品行业经济的健康发展具有重要意义。利用同步荧光光谱对掺杂牛奶进行检测,寻求一种高效判别掺杂乳品方法。采用分子荧光分光光度计测定激发波长(Ex)为220~600nm,激发-发射间隔波长(Δλ)为10~180nm的纯牛奶、复原乳粉(全脂、脱脂)及其掺杂样本的牛奶样品的三维荧光光谱数据,利用平行因子分析方法(PARAFAC)降维获取特征光谱,通过支持向量机建立了掺杂牛奶的判别模型。结果所有乳品样品在激发波长为225~300nm范围内都有一个特征荧光峰,荧光峰在280nm附近,为色氨酸类物质,但该处纯牛奶荧光强度明显高于两种乳粉,且脱脂乳粉要强于全脂乳粉,这说明牛奶的主要成分都一样,发光基团一致,但由于处理方式不一致,使得其浓度存在差异。两种复原乳粉在350~400,450~500nm之外存在荧光物质,主要为维生素A和类胡萝卜素,且脱脂乳粉比全脂乳粉对应区域荧光强度要强,主要在于脂肪物质散射使得荧光强度增强。为更好获取乳品样本特征,通过PARAFAC对三维数据进行降维之后,显示当组分数为6,Δλ为40nm时载荷值最大,该处样本信息差异显著。提取Δλ为40nm特征波长和掺杂乳品品类值作为输入数据的支持向量机(SVM)分类器,采用了遗传算法(Ga-SVM)、粒子群优化算法(Pso-SVM)和网格搜索算法(Grid-SVM)三种SVM算法对掺杂牛奶进行分类识别。结果显示Grid-SVM模式交叉验证(CV)准确率为98.91%,其训练集和测试集的分类准确率均为100.00%,且模型运行时间仅6.724s,显著优于另两种分类器。结果表明荧光光谱与PARAFAC-SVM方法相结合,是一种简单且高效判别掺杂牛奶的方法。

二维相关近红外光谱的牦牛奶粉掺伪判别分析

目的:建立二维相关近红外光谱结合偏最小二乘判别分析方法(PLS-DA)对牦牛奶粉掺伪进行快速判别分析。方法:通过将牦牛奶粉与普通牛奶粉以一定比例混合,共制备56个掺伪样品,并以KS法按3∶1比例划分校正集和预测集;再采集所有样品一维近红外光谱,构建出以浓度为外扰的二维相关谱,基于同步谱与自相关谱的分析,优选出26个特征变量,建立牦牛奶粉掺伪PLS-DA判别模型。结果:通过二维相关光谱分析,优选到的26个特征波长,集中在光谱差异比较明显的区域,均与奶粉中的蛋白质脂肪等营养成分密切相关;建立的牦牛奶粉掺伪判别模型,校正集与预测集的决定系数R^(2)均超过0.97,校正集与预测集的分类准确率均为100%。结论:结果表明,二维相关光谱结合化学计量学方法可以识别掺伪牦牛奶粉的微小差异特征,并进行快速的判别,本方法为乳制品掺伪检测提供了新的思路和方法。

应用Milkoscan FT 120对牛奶掺假的研究

采用红外扫描乳品测定仪对原料乳中脂肪、蛋白、干物质、乳糖、柠檬酸密度、冰点、碳水化合物、全糖、游离脂肪酸等成分参数的检测建立原料乳光谱数据库,在具有代表性原料乳中加入植脂末模拟牛奶掺假,通过应用掺假模块鉴别生鲜牛奶是否掺假。

Kinetic Determination of Urea in Milk Powder by Nonlinear Chemical Fingerprint Technique

To detect urea content in milk powder, a nonlinear chemical fingerprint technique was developed. In our study, Belousov-Zhabotinsky oscillatory chemical reaction (B-Z) was performed using milk powder and malonic acid as main dissipative substances. The same dosage of milk powder with or without artificially added urea was introduced to “H+ + Ce4+ + + malonic acid” oscillating system, respectively, and nonlinear chemical fingerprints of different milk powder were thus obtained. The proposed method was based on a linear relationship between inductive time of non-linear chemical fingerprints and urea content in milk powder, which held when urea content in milk powder was in the range of 0 - 40 mg/g. A detection limit of 7.8 × 10-3mg/g was also obtained. Our results showed that the method could be used to certify urea content in milk powder without pretreatment of samples, which was more simple and economical compared with traditional methods.

Hypoallergenicity of a thickened hydrolyzed formula in children with cow’s milk allergy

BACKGROUND Allergy to cow’s milk is the most frequent allergy occurring in infants and young children.The dietary management of these patients consists of the elimination of any cow’s milk proteins from the diet,and for formula-fed infants,the substitution of the usual infant formula with an adapted formula that is generally based on extensively hydrolyzed cow’s milk proteins.The American Academy of Pediatrics has established specific criteria to confirm the hypoallergenicity of a formula intended for these children.AIM To assess the hypoallergenicity of a new thickened extensively hydrolyzed casein-based formula (TeHCF) in children with cow’s milk allergy (CMA).METHODS Children diagnosed with CMA through a double-blind placebo-controlled food challenge (DBPCFC) were randomly administered increased doses of a placebo formula or the TeHCF [Allernova,new thickener including fibres (Novalac)] under double-blind conditions and medical surveillance on two separate days.Otherwise,both of these formulas and a cow’s milk-based formula were randomly introduced to children who were highly suspected of having CMA on three separate days.Immediate and late reactions occurring after the introduction of any of these formulas were thoroughly recorded by the physician at the hospital and reported by parents to the physician after hospital discharge,respectively.If the children tolerated the TeHCF during the DBPCFC,they were exclusively fed this formula during a 3-mo period where potential allergic symptoms,anthropometric parameters,as secondary outcomes,and adverse events were registered.The Cow’s Milk-related Symptoms Score (CoMiSSTM) was assessed and anthropometric parameters were compared to World Health Organization (WHO) reference data.RESULTS Of the 30 children included in the study,the CMA diagnosis of 29 (mean age: 8.03 ± 7.43 mo) patients was confirmed by a DBPCFC.The children all tolerated the TeHCF during both the challenge and the subsequent 3-mo feeding period,which they all completed.During the latter period,the CoMiSSTM remained at a very low level,never exceeding its baseline value (1.4 ± 2.0),growth parameters were within WHO reference standards and no adverse event related to the TeHCF was reported.Over the first week of this period,the proportion of patients with digestive discomfort significantly decreased from 20.7%(6/29) to 3.4%(1/29),P = 0.025.The proportion of satisfaction with the overall effect of the formula reported by the parents and investigator was high,as was the formula acceptability by the child.CONCLUSION The new TeHCF meets the hypoallergenicity criteria according to the American Academy of Pediatrics standards,confirming that the tested TeHCF is adapted to the dietary management of children with CMA.Moreover,growth was adequate in the included population.

Detection of Glucomacropeptide in Raw Milk Adulterated with Cheese Whey in Ecuador

Milk is one of the products that can be adulterated in many ways affecting the quality of this and its derivatives. Glucomacropeptide (GMP) is a protein that is found only in the whey from the production of fresh cheese, enzymatically obtained from the coagulation of casein and which is commonly used to adulterate fresh or powdered milk. The aim of this study was to determine the adulteration of milk with cheese whey thought a molecular approach, where the glucomacropeptide was collected by sequential precipitation with trichloroacetic acid (ATC) and detected by polyacrylamidododecylsulfate gel electrophoresis (PAGE-SDS), using samples of fresh milk, intentionally adulterated with serum in the proportion of 0%, 1%, 5%, 10% and 15%. The results obtained showed that the detection of glucomacropeptide by electrophoresis was positive in all samples of adulterated milk, evidencing a band of 20.9 kDa in the reading, corresponding to the molecular weight of the GMP, showing that the technique used determines the adulteration in the milk, in a specific and sensitive way, also shows that in the evaluation of physical-chemical and microbiological parameters of milk, there are no significant differences between treatments, except for the pH that tends to decrease as the percentage of serum in the milk increases.

牛、羊乳粉的DSC热学性质比较及掺假分析

为明确羊乳粉、牛乳粉的热学特征,采用差示扫描量热法(differential scanning calorimetry,DSC)对真空冷冻干燥制备的全脂羊、牛乳粉样品以及高比例掺假(75%、50%、25%)和低比例掺假(10%、5%、3%、1%)牛乳粉的混合羊乳粉样品进行热力学分析。结果表明,全脂羊乳粉和全脂牛乳粉在DSC热学指标上存在差异,全脂牛乳粉相比全脂羊乳粉缺失一个脂肪特征熔融吸热峰b,蛋白质熔融吸热峰c峰值温度和热焓值较低,而乳糖熔化分解峰e热焓值较高。对于掺入不同比例牛乳粉的羊乳粉,通过检测是否存在吸热峰b及其热焓值,可判断样品掺假牛乳粉比例是否在25%以下及判断掺入牛乳粉的量;在不同比例掺假样品中检测乳糖熔化分解峰e的焓值可判断羊乳粉掺入牛乳粉的掺假量。因此,DSC技术可以实现对羊乳粉、牛乳粉热学性质的分析和评价,也可作为乳制品行业质量保证和真实性鉴别的潜在分析工具。

非变性聚丙烯酰胺凝胶电泳检测新疆特种乳中掺加的牛乳

新疆特种乳资源丰富,具有重要的商业价值。近年来乳制品掺假问题时有发生,准确鉴别特种乳中乳源动物成分,防止牛乳掺假具有重要的意义。本研究采用非变性聚丙烯酰胺凝胶电泳(native-PAGE)对驼乳、马乳、驴乳、山羊乳与牛乳等特种乳的乳清蛋白及酪蛋白进行乳间差异性分析,建立基于牛乳特征蛋白质多态性的检测方法,并研究不同热处理工艺对检测方法稳定性、灵敏度的影响。结果表明,牛β-乳球蛋白可作为特种乳中掺加牛乳的检测靶标,热处理会使样品中蛋白质发生变性,导致掺加牛乳检出限的升高。基于牛β-乳球蛋白的条带强度与牛乳掺加百分比建立回归方程,相关系数为0.9779~0.9998,定量范围在5%~100%之间。本研究为新疆特种乳中掺加牛乳的检测,提供了一种准确、灵敏和经济的方法。