In situ imaging highlights local structural changes during heating: the case of meat

Understanding and monitoring deformation and water content changes in meat during cooking is of prime importance. We show the possibilities offered by nuclear magnetic resonance imaging (MRI) for the in situ dynamic measurement of deformation fields and water content mapping during beef heating from 20 to 75 °C. MRIs were acquired during heating, and image registration was used to calculate the deformation field. The temperature distribution in the sample was simulated numerically to link structural modifications and water transfer to temperature values. During heating, proton density decreases because of a magnetic susceptibility drop with temperature and water expulsion due to muscle contraction. A positive relationship was found between local cumulative deformation and water content. This new approach makes it possible to identify the deformation field and water transfer simultaneously and to trace thermal history to build heuristic models linking these parameters.     

基于LF-NMR研究包装方式和温度对鲶鱼片保水性的影响

为了考察贮藏过程中包装方式和温度对鲶鱼片保水性的影响,该试验将新鲜鲶鱼片分别经空气包装(air-package,AP)、真空包装(vacuum package,VP)和气调包装(modified atmosphere package,MAP,60%CO_2+40%N_2)后贮藏于4℃和-0.7℃的冷库中,于贮藏第0、1、4、7、10、15、20、30天测定鲶鱼片的pH值、蒸煮损失率、离心失水率等指标变化,利用低场核磁共振(low-field nuclear magnetic resonance,LF-NMR)技术测定鲶鱼片的水分弛豫时间(T22)、弛豫面积(P22)和质子密度成像(magnetic resonance imaging,MRI),并利用扫描电镜(scanning electron microscopy,SEM)观察鲶鱼片肌纤维结构的变化,对鲶鱼片贮藏过程中保水性变化及原因进行综合判断。结果显示,随着贮藏时间的延长,各组鲶鱼片保水性均呈下降趋势,表现为蒸煮损失率和离心失水率比新鲜鲶鱼片增加,弛豫时间T22和弛豫面积P22降低。相比较而言,冰温气调(-0.7°C-MAP)样品的保水性下降最为缓慢,是鲶鱼片品质保持的较佳贮藏方法。皮尔逊相关系数分析表明弛豫面积P22与贮藏时间、蒸煮损失率及离心失水率显著相关(P0.05);MRI结果显示,鲶鱼片的水分在贮藏过程中逐渐从肌纤维内部渗出,集聚在肌束膜上;SEM表明肌纤维细胞结构发生改变,这可能是引起保水性下降的根本原因。综上所述,该研究证明利用LF-NMR技术可快速表征鲶鱼肉保水性的变化,研究结果为选择鲶鱼片的较佳贮藏条件提供了理论依据。     

Heat-induced changes in myofibrillar protein structures and myowater of two pork qualities. A combined FT-IR spectroscopy and low-field NMR relaxometry study

Low-field NMR T(2) and Fourier transform infrared (FT-IR) measurements were performed on meat samples of two qualities (normal and high ultimate pH) during cooking from 28 degrees C to 81 degrees C. Pronounced changes in both T(2) relaxation data and FT-IR spectroscopic data were observed during cooking, revealing severe changes in the water properties and structural organization of proteins. The FT-IR data revealed major changes in bands in the amide I region (1700-1600 cm(-)(1)), and a tentative assignment of these is discussed. Distributed NMR T(2) relaxation data and FT-IR spectra were compared by partial least-squares regression. This revealed a correlation between the FT-IR peaks reflecting beta-sheet and alpha-helix structures and the NMR relaxation populations reflecting hydration water (T(2B) approximately 0-10 ms), myofibrillar water (T(21) approximately 35-50 ms), and also expelled "bulk" water (T(2) relaxation times >1000 ms). Accordingly, the present study demonstrates that definite structural changes in proteins during cooking of meat are associated with simultaneous alterations in the chemical-physical properties of the water within the meat.     

Cooking temperature is a key determinant of in vitro meat protein digestion rate: investigation of underlying mechanisms

The present study aimed to evaluate the digestion rate and nutritional quality of pig muscle proteins in relation to different meat processes (aging, mincing, and cooking). Under our experimental conditions, aging and mincing had little impact on protein digestion. Heat treatments had different temperature-dependent effects on the meat protein digestion rate and degradation potential. At 70 °C, the proteins underwent denaturation that enhanced the speed of pepsin digestion by increasing enzyme accessibility to protein cleavage sites. Above 100 °C, oxidation-related protein aggregation slowed pepsin digestion but improved meat protein overall digestibility. The digestion parameters defined here open new insights on the dynamics governing the in vitro digestion of meat protein. However, the effect of cooking temperature on protein digestion observed in vitro needs to be confirmed in vivo.     

Influence of aging and salting on protein secondary structures and water distribution in uncooked and cooked pork. A combined FT-IR microspectroscopy and 1H NMR relaxometry study

Fourier transform infrared (FT-IR) microspectroscopy and low-field (LF) proton NMR transverse relaxation measurements were used to study the changes in protein secondary structure and water distribution as a consequence of aging (1 day and 14 days) followed by salting (3%, 6%, and 9% NaCl) and cooking (65 degrees C). An enhanced water uptake and increased proton NMR relaxation times after salting were observed in aged meat (14 days) compared with nonaged meat (1 day). FT-IR bands revealed that salting induced an increase in native beta-sheet structure while aging triggered an increase in native alpha-helical structure before cooking, which could explain the effects of aging and salting on water distribution and water uptake. Moreover, the decrease in T2 relaxation times and loss of water upon cooking were attributed to an increase in aggregated beta-sheet structures and a simultaneous decrease in native protein structures. Finally, aging increased the cooking loss and subsequently decreased the final yield, which corresponded to a further decrease in T2 relaxation times in aged meat upon cooking. However, salting weakened the effect of aging on the final yield, which is consistent with the increased T2 relaxation times upon salting for aged meat after cooking and the weaker effect of aging on protein secondary structural changes for samples treated with high salt concentration. The present study reveals that changes in water distribution during aging, salting, and cooking are not only due to the accepted causal connection, i.e., proteolytic degradation of myofibrillar structures, change in electrostatic repulsion, and dissolution and denaturation of proteins, but also dynamic changes in specific protein secondary structures.     

Monitoring of denaturation processes in aged beef loin by Fourier transform infrared microspectroscopy

We present the results of a Fourier transform infrared (FT-IR) microspectroscopic study using conventional FT-IR microscopy and FT-IR imaging to detect the denaturation process during four different heating temperatures (raw, 45, 60, and 70 degrees C) spatially resolved in bovine cryosections from longissimus dorsi muscle. FT-IR imaging, employing a focal plane array detector, which allowed the simultaneous collection of spectra at 4096 pixels, enabled the investigation of the heat-induced changes in the two major meat constituents, i.e., myofibrillar and connective tissue proteins, spatially resolved. The infrared spectra of both compounds revealed that the major spectral changes involved an increase in beta-sheet and a decrease in alpha-helical structures, which appeared to be much more pronounced for the myofibers than for the connective tissue. These conformational changes could be correlated to the denaturation of the major meat proteins, such as myosin, actin, and collagen.     

Myowater dynamics and protein secondary structural changes as affected by heating rate in three pork qualities: a combined FT-IR microspectroscopic and 1H NMR relaxometry study

The objective of this study was to investigate the influence of heating rate on myowater dynamics and protein secondary structures in three pork qualities by proton NMR T2 relaxation and Fourier transform infrared (FT-IR) microspectroscopy measurements. Two oven temperatures at 100 degrees C and 200 degrees C corresponding to slow and fast heating rates were applied on three pork qualities (DFD, PSE, and normal) to an internal center temperature of 65 degrees C. The fast heating induced a higher cooking loss, particularly for PSE meat. The water proton T21 distribution representing water entrapped within the myofibrillar network was influenced by heating rate and meat quality. Fast heating broadened the T21 distribution and decreased the relaxation times of the T21 peak position for three meat qualities. The changes in T21 relaxation times in meat can be interpreted in terms of chemical and diffusive exchange. FT-IR showed that fast heating caused a higher gain of random structures and aggregated beta-sheets at the expense of native alpha-helixes, and these changes dominate the fast-heating-induced broadening of T21 distribution and reduction in T21 times. Furthermore, of the three meat qualities, PSE meat had the broadest T21 distribution and the lowest T21 times for both heating rates, reflecting that the protein aggregation of PSE caused by heating is more extensive than those of DFD and normal, which is consistent with the IR data. The present study demonstrated that the changes in T2 relaxation times of water protons affected by heating rate and raw meat quality are well related to the protein secondary structural changes as probed by FT-IR microspectroscopy.     

Influence of the spatial organization of the perimysium on beef tenderness

The spatial distribution of the intramuscular connective tissue (IMCT) in four types of beef muscle (Biceps femoris, Infraspinatus, Longissimus thoracis, and Pectoralis profundus) was examined using histology and magnetic resonance imaging (MRI). The surface and the length of the IMCT and the surface of the myofiber bundles were evaluated by image analysis. The texture of the cooked meat from these muscles was measured both instrumentally by a compression test and by sensory analysis. The relationship between muscle structure and meat texture was studied by general discriminant analysis. The models obtained could assign correctly up to 87% of the samples to two tenderness classes. Histology and MRI provided complementary information about the microscopic and macroscopic IMCT structures, respectively. Both were necessary to predict sensory tenderness whereas only the MRI measurements were necessary to predict instrumental toughness. Tough muscles had smaller MRI myofiber bundles (0.7-1 mm radius) than tender muscles.     

Cooking effects on water distribution in potatoes using nuclear magnetic resonance relaxation

Continuous low-field (LF) (1)H NMR relaxometry was used to monitor the structural changes during cooking of potatoes with two different dry matter (DM) contents. A principal component analysis of the relaxation decay curves revealed major events related to water mobility during cooking, which occur at 53 and 60 degrees C for potatoes with medium and low DM contents, respectively. Exponential analysis of the relaxation decays reveals two major water populations in the potato: a slow-relaxing (assigned to water in cytoplasm and extracellular cavities) water component, T(22) ( approximately 350-550 ms), and a fast-relaxing component (primarily assigned to water associated with starch and cell walls), T(21) ( approximately 45-65 ms). Significant DM dependent shifts in both the T(21) and T(22) relaxation time constants were observed during cooking, indicating that starch gelatinizes between 53 and 70 degrees C with water exchanging with the hydroxyls of starch (transition in T(21)) and cells start to disrupt with an increase in diffusion volumes at approximately 60 degrees C (transition in T(22)). The study reveals that continuous LF NMR measurement is an excellent and highly sensitive method to study changes in water mobility and water populations during the cooking of potatoes.     

Salt diffusion and distribution in meat studied by 23Na nuclear magnetic resonance imaging and relaxometry

This study introduces the use of combined 23Na magnetic resonance imaging (MRI) and 23Na NMR relaxometry for the study of meat curing. The diffusion of sodium ions into the meat was measured using 23Na MRI on a 1 kg meat sample brined in 10% w/w NaCl for 3-100 h. Calculations revealed a diffusion coefficient of 1 x 10(-5) cm2/s after 3 h of curing and subsequently decreasing to 8 x 10(-6) cm2/s at longer curing times, suggesting that changes occur in the microscopic structure of the meat during curing. The microscopic mobility and distribution of sodium was measured using 23Na relaxometry. Two sodium populations were observed, and with increasing length of curing time the relaxation times of these changed, reflecting a salt-induced swelling and increase in myofibrillar pore sizes. Accordingly, the present study demonstrated that pore size and thereby salt-induced swelling in meat can be assessed using 23Na relaxometry.     

考虑收缩的爆炒热/质传递过程模拟与验证

为研究爆炒中食品多孔介质热/质传递机制及烹饪成熟和品质变化规律，考虑收缩-水分损失关系，基于多孔介质理论，结合傅里叶定律、牛顿冷却定律和达西定律，构建了爆炒中有蒸发、考虑收缩的食品含湿非饱和多孔介质热/质传递数学模型，开展了爆炒数值模拟。考虑收缩后模拟值与实测温度历史（Least Summation of the squared Temperature Difference for overall target，LSTD=4.40 ℃）、平均含水率（LSTD=1.42%）和体积收缩率（LSTD=1.05%）吻合更为良好。模拟分析了爆炒热/质传递对颗粒表面蒸发、收缩、内部压力、水分和温度分布的影响机理及火候控制手段的作用，结果表明：爆炒强对流传热使颗粒蒸发剧烈造成水分损失；收缩主要由水分损失引起，可以增大传热效率并影响颗粒内部压力变化，由成熟值理论研判，蒸发收缩对烹饪成熟起到促进作用，并有利于提高烹饪品质，是爆炒技术优势的一部分；火候控制手段通过改变颗粒特征尺寸、能量传递速率和流体-颗粒的换热时间/接触面积对颗粒成熟时间和含水率等烹饪品质产生极显著（P<0.01）影响。     

NO-AMPK通路对牛肉宰后成熟过程中蛋白特性与肉品质的影响

为研究宰后NO-AMPK通路对牛肉蛋白特性及肉品质的影响,以一氧化氮合成酶(NOS)抑制剂L-NAME、一磷酸腺苷活化蛋白激酶(AMPK)抑制剂Compound C处理的牛臀股四头肌为研究对象,生理盐水处理作为对照,将肉样与处理液在4℃条件下1:1(g:mL)混合浸泡12 h后,在成熟0、6、12、24、48、72、120 h测定肉样的蛋白功能特性、保水性、嫩度以及肌肉组织结构等相关指标。结果显示,L-NAME组中一氧化氮NO含量在6~72 h显著低于对照组(P<0.05);3组中AMPK活性先增大后减小,L-NAME组和Compound C组中AMPK活性在6 h分别比对照组降低14.78%和26.75%;L-NAME组中总蛋白溶解性在6、24 h低于Compound C组,高于对照组(P<0.05),表面疏水性在12、48 h显著低于对照组(P<0.05)并高于Compound C组;L-NAME组中剪切力仅在48 h显著高于对照组并低于Compound C组(P<0.05);对照组中肉样蒸煮损失最大,Compound C组中肉样蒸煮损失最小。以上结果表明,牛肉宰后成熟过程中,NO能够通过AMPK通路改善牛肉嫩度,但对其保水性产生了不利影响。本研究结果可为牛肉宰后能量代谢及肉品质控制相关研究提供一定的理论基础。     

Effects of Milling and Cooking Conditions of Rice on In Vitro Starch Digestibility and Blood Glucose Response

The objective of this study was to investigate the effects of milling and cooking conditions of cooked rice prepared from cultivar Koshihikari on in vitro starch digestibility and in vivo glucose response in humans. In addition, compression and adhesiveness tests were conducted for texture analysis of the cooked rice. Brown rice (BR) and surface‐abraded BR (SABR, ≥99.5% of the original weight) were digested more slowly than white rice (91% of the original weight) when cooked rice grain was used for the in vitro test, but they were digested more rapidly in the initial stage of the reaction when cooked rice ground by a meat grinder was used. The increase in water added for cooking significantly increased the extent of starch digestion with BR and SABR. The changes in blood glucose levels after the ingestion of cooked rice were dependent on the sample type. The cooking conditions dramatically influenced the glucose response after the ingestion of BR. A significant correlation was found between blood glucose levels at 45 min and the extent of starch digestion with ground samples, whereas no relationship was found with cooked rice grain samples for in vitro digestibility.     

Headspace oxygen in sample vials affects volatiles production of meat during the automated purge-and-Trap/GC analyses.

Headspace oxygen in sample vial for the purge-and-trap dynamic headspace/gas chromatography method oxidizes meat if held hours before purging, influences volatile profiles, and misrepresents the true composition of volatiles. Helium flush and helium flush plus oxygen absorber were used to eliminate residual oxygen and minimize oxidative changes in meat during sample holding time. Both helium flush and helium flush plus oxygen absorber treatments were effective in preventing an increase in 2-thiobarbituric acid reactive substances (TBARSs) and volatiles production in raw meat for up to 640 min of sample holding. With helium flush plus oxygen absorber, only 1-octen-3-ol increased during the 1280-min sample holding time. However, the hexanal peak in raw meat was interfered by 2,6-dimethyl heptane when oxygen absorber was added. Therefore, use of oxygen absorber was not appropriate for raw meat. Helium flush reduced oxidative changes in cooked meat during sample holding time but was not able to stop oxidative changes in meat after 160 min sample holding. A combination of helium flush and oxygen absorber was effective in preventing volatiles production in cooked meat for over 20 h of sample holding at 4 degrees C.     

Morphological examinations of oxidatively stressed pork muscle and myofibrils upon salt marination and cooking to elucidate the water-binding potential

Pork longissimus muscle samples were subjected to the following three marination conditions: (A) oxidation (40 min) in hydroxyl radical-generating solutions (HRGS; 10 μM FeCl(3)/100 μM ascorbate with 5 or 20 mM H(2)O(2), pH 6.2) containing 0.1 M NaCl and then marination (40 min) in 0.6 M NaCl with 15 mM pyrophosphate (PP); (B) simultaneous oxidation/marination (40 min) in HRGS containing 0.6 M NaCl and 15 mM PP; or (C) the same as condition B except that PP was omitted. Protein oxidation, measured by the carbonyl and tryptophan fluorescence changes, enhanced hydration but increased cooking loss of meat. Light microscopy revealed a dense muscle structure characterized by swollen fibers and reduced intercellular spacing in intermediately oxidized muscle samples marinated with 0.6 M NaCl and 15 mM PP. However, oxidized fibers were more susceptible to transverse shrinkage upon cooking than nonoxidized fibers, which was supported by the dynamic ultrastructural changes in myofibrils observed using phase contrast microscopy. These findings provide a further understanding of the complex impact of oxidation on meat hydration and water-binding.     

Effect of the polyunsaturated fatty acid composition of beef muscle on the profile of aroma volatiles

The effect of n-3 polyunsaturated fatty acids (PUFAs) in beef muscle on the composition of the aroma volatiles produced during cooking was measured. The meat was obtained from groups of steers fed different supplementary fats: (i) a palm-oil-based control; (ii) bruised whole linseed, which increased muscle levels of alpha-linolenic (C18:3 n-3) and eicosapentaenoic acid (EPA, C20:5 n-3); (iii) fish oil, which increased EPA and docosahexaenoic acid (C22:6 n-3); (iv) equal quantities of linseed and fish oil. Higher levels of lipid oxidation products were found in the aroma extracts of all of the steaks with increased PUFA content, after cooking. In particular, n-alkanals, 2-alkenals, 1-alkanols, and alkylfurans were increased up to 4-fold. Most of these compounds were derived from the autoxidation of the more abundant mono- and di-unsaturated fatty acids during cooking, and such autoxidation appeared to be promoted by increased levels of PUFAs.     

新铁炮百合鳞茎低温解除休眠过程中的形态和生理变化

以自繁的新铁炮百合鳞茎为材料,研究在4°C、8°C和12°C低温处理下鳞茎解除休眠过程中的形态和生理变化,结果表明,百合鳞茎在冷藏过程中其顶芽和新根均不断伸长,其伸长速度排序为12°C处理>8°C处理>4°C处理,在8°C和12°C下冷藏37 d其顶芽生长点距离鳞茎顶端约1 cm。在45 d的冷藏期内,各冷藏处理的鳞茎的淀粉含量明显下降;可溶性蛋白质含量呈下降趋势,但冷藏30~37 d有明显升高过程;总可溶性糖含量和还原糖含量都呈先上升后下降的趋势,冷藏37 d时两者均达到峰值,然后出现下降,糖分含量变化的转折点与鳞茎解除休眠有关。在本试验中,以冷藏37 d为新铁炮百合鳞茎解除休眠的合适时间。     

低压静电场下不同隔距冻结-解冻对牛肉品质的影响

为研究低压静电场(low voltage electrostatic field,LVEF)辅助冻结-解冻对牛肉品质的影响,采用牛背最长肌作为试验材料,探究与静电场发生板隔距分别为15、30、45、60 cm处(试验组,冻结温度为?18℃,解冻温度为4℃)和自然冻结-解冻(未施加静电场,对照组,条件同上)肉样冻结-解冻过程中的品质变化。对比分析了温度曲线、色泽、解冻汁液流失、汁液中蛋白含量、蒸煮损失及质构特性等指标变化,显微观察肌肉纤维组织中的冰晶形态和肌肉微观结构。结果表明:与对照相比,试验组牛肉在冻结过程中通过最大冰晶生成带时间较短,组织中的冰晶较对照组体积小、数目多、分布均匀;肉样解冻速度较快,解冻后肉色更新鲜,咀嚼性、嫩度较高,解冻汁液流失率、汁液中蛋白含量及蒸煮损失率均显著降低(P0.05);扫描、透射电镜结果表明,试验组肉样解冻后肌肉微观结构遭破坏程度较轻,肌纤维束排列相对紧密。其中,与静电板隔距为30 cm处冻结-解冻的肉样亮度值、红度值和色彩饱和度值分别为39.47、21.77和23.71,显著高于对照组的31.74、17.76和20.73(P0.05);解冻汁液流失率、蒸煮损失率及解冻汁液中蛋白质量分数较对照分别降低4.18、8.28、0.7个百分点,咀嚼性增加32.68 N,差异显著(P0.05)。试验结果表明低压静电场辅助能显著缓解牛肉在冻结和解冻过程中的品质劣变,提高解冻牛肉品质,肉样在与静电板隔距为30 cm处冻结-解冻较为适宜。     

Detection and localization of oxidized proteins in muscle cells by fluorescence microscopy

In meat, no detailed studies on the intracellular distribution of oxidized proteins during oxidative stress have been performed, to our knowledge. Therefore, we used fluorescence microscopy to detect and locate protein carbonyls, oxidation products of basic amino acids, generated in bovine M. Rectus abdominis during either exposition to a chemical free radical generating system, or refrigerated storage, or cooking. The technique consisted of an immunohistochemical detection of carbonyls by reaction with the specific probe DNPH (2,4-dinitrophenylhydrazine) followed by the sequential addition of a first antibody against DNPH-carbonylated proteins and a CY3-labeled secondary antibody. The fluorescence of the CY3 probe increased regularly with level of free radical generating system and storage time. Moreover, an important heterogeneity of carbonyl distribution was observed, with a higher oxidation level at the periphery than inside the muscle cells. Cooking induced fluorescence increase only at the periphery of cells. Specific coloration of collagen by Sirius red showed that collagen was not involved in fluorescence. We can deduce that accumulation of oxidized proteins observed in the cell periphery was linked to membrane protein oxidation and not to connective tissue oxidation. Biochemical assays were performed in parallel on membrane and myofibrillar proteins to provide complementary quantitative data on level of oxidized proteins.     

中式烹饪用时间温度积分器的构建与验证

蛋白质变性能够较广泛地表征烹饪加热品质变化,因而寻找到一种z值为7.36℃的耐高温α-淀粉酶,与蛋白质热变性z值5~10℃相近。以该酶溶液为指示剂,在玻璃毛细管中封装后置入烹饪耐受性高的、特定形状的魔芋凝胶(g-KGM)载体,从而构建了烹饪研究用时间温度积分器(time temperature integrators,TTIs)装置。随后,在模拟烹饪过程而设定的对流传热条件下,通过传热学试验结合非稳态传热以及酶失活动力学数学模型计算得到剩余酶活,与TTIs装置指示剂酶活实测值比较,两者误差小于2.24%。进一步,应用该TTIs装置测定了实际烹饪爆炒过程的表面换热系数。所构建的TTIs装置,结合数值模拟,可以分析测量常规试验传热学方法无法应用的激烈烹饪中流体-颗粒的传热过程,也可应用于其他领域的移动颗粒传热学研究。