Scallops as a new source of food protein: high-intensity ultrasonication improved stability of oil-in-water emulsion stabilised by myofibrillar protein

In this study, the effect of high-intensity ultrasound (HIUS) (200 and 400 W for 0, 5, 10 and 15 min respectively) on conformational changes, physicochemical, rheological and emulsifying properties of scallop (Patinopecten yessoensis) myofibrillar protein (SMP) was investigated. HIUS-treated SMP had lower α-helix content and higher β-sheet content compared with the native SMP. HIUS treatment induced the unfolding of SMP and increased the surface hydrophobicity. The particle size of SMP decreased and the absolute zeta-potential increased after ultrasonication, which in turn increased the solubility of SMP. The conformational changes and the improvement of physicochemical properties of SMP increased the ability for SMP to lower the interfacial tension at the oil–water interface and increased the percentage of adsorbed protein. As a result, the emulsifying properties, rheological properties of SMP and storage stability of emulsions were also improved. In conclusion, HIUS treatment has future potential for improving the emulsifying properties of SMP.     

Evaluation of physicochemical properties and microbial counts of raw and cooked low-fat patties added with eggplant powder prepared with different drying methods

Effects of different drying methods and different addition levels of eggplant (EP) on product quality of low-fat patties (LFPs) were investigated during storage. EP was dried in an oven dryer at 60 °C or a freeze dryer at −50 °C. LFPs were prepared by replacing with 1.5% soy protein isolate (SPI). Six treatments were used in this study: (1) control (CTL), without addition of EP; (2) reference (REF), 0.1% ascorbic acid; (3) O1, 0.25% oven-dried (OD) EP; (4) O2, 0.5% ODEP; (5) F1, 0.25% freeze-dried (FD) EP; and (6) F2, 0.5% FDEP. Redness (a*) and lightness (L*) values in LFPs added with EP were lower than those of others (p < 0.05) and decreased with increasing storage time. Yellowness (b*) values of cooked patties were increased during storage time (p < 0.05), with control having the highest value. The addition of EP or ascorbic acid into LFPs lowered microbial counts than control (p < 0.05). Thiobarbituric acid reactive substances (TBARS) was increased during storage, with REF having the lowest value, and patties added with EP had lower TBARS values than control during storage. Volatile basic nitrogen (VBN, mg%) contents of all patties also increased during storage time with O2 and F2 having lower values than control. Therefore, EP might have potential as a natural antioxidant in meat products during storage.     

具有保护肠上皮细胞HT-29免受大肠杆菌和H2O2损伤潜力的植物乳杆菌的筛选

益生菌可在肠道定植从而发挥抗炎或抗氧化活性，有利于宿主肠道健康。本实验研究了从新疆传统发酵乳制品中分离得到的8?株植物乳杆菌对大肠杆菌侵袭和过氧化氢刺激肠上皮细胞HT-29的保护作用。结果表明：在8?株植物乳杆菌中，植物乳杆菌35具有最高的黏附能力。植物乳杆菌35可通过取代、竞争、排阻的方式抑制大肠杆菌对HT-29细胞的黏附，抑制率分别为42.60%、59.17%、60.19%。植物乳杆菌35及其多糖可抑制大肠杆菌刺激HT-29细胞产生白细胞介素-8；同时保护HT-29细胞免受过氧化氢的损伤，增加超氧化物歧化酶、谷胱甘肽过氧化物酶活力水平并降低丙二醛含量。结论：植物乳杆菌35及其粗胞外多糖具有抑制大肠杆菌O157诱导的炎症性肠病的潜力。     

Polysorbate identity and quantity dictate the extensional flow properties of protein-excipient solutions

While protein medications are promising for treatment of cancer and autoimmune diseases, challenges persist in terms of development and injection stability of high-concentration formulations. Here, the extensional flow properties of protein-excipient solutions are examined via dripping-onto-substrate extensional rheology, using a model ovalbumin (OVA) protein and biocompatible excipients polysorbate 20 (PS20) and 80 (PS80). Despite similar PS structures, differences in extensional flow are observed based on PS identity in two regimes: at moderate total concentrations where surface tension differences drive changes in extensional flow behavior, and at small PS:OVA ratios, which impact the onset of weakly elastic flow behavior. Undesirable elasticity is observed in ultra-concentrated formulations, independent of PS identity; higher PS contents are required to observe these effects than in analogous polymeric excipient solutions. These studies reveal novel extensional flow behaviors in protein-excipient solutions, and provide a straightforward methodology for assessing the extensional flow stability of new protein-excipient formulations.     

四氢姜黄素经PI3K/Akt信号通路对人血小板活化和聚集的调控作用

目的：探讨姜黄素的主要肠道代谢物四氢姜黄素（tetrahydrocurcumin，THC）对血小板活化和聚集的影响及其可能的分子机制。方法：在体外实验中，用不同浓度的THC（0、0.5、1、10 μmol/L）提前与健康人纯化血小板共同孵育40 min，然后加入凝血酶激活血小板2 min，用流式细胞术测定血小板表面CD62P和CD63的表达量，用酶联免疫吸附法测定血小板释放血小板因子-4（platelet factor-4，PF4）和趋化因子配体-5（chemokine ligand 5，CCL5）水平，用血小板聚集仪检测血小板释放ATP水平和血小板最大聚集率，用Western blot蛋白免疫印迹法检测血小板磷酸肌醇-3-激酶（phosphoinositide 3-kinase，PI3K）和Akt蛋白的磷酸化水平。结果：与模型组（血小板悬液中加入0.05%二甲基亚砜）相比，THC能抑制凝血酶诱导的血小板表面CD62P和CD63的表达，抑制PF4、CCL5和ATP的释放，降低血小板最大聚集率，下调PI3K和Akt蛋白的磷酸化水平，且呈浓度依赖效应，其中10 μmol/L的浓度下作用效果显著（P＜0.01、P＜0.001）。PI3K的特异性激动剂740 Y-P可部分逆转THC对PF4和CCL5释放和血小板聚集的抑制作用（P＜0.05、P＜0.01）。结论：THC具有显著抑制血小板活化和聚集的作用，其机制可能是THC可下调PI3K/Akt介导的信号通路。     

Effects of multiple freeze–thaw cycles on meat quality,nutrients, water distribution and microstructure in bovine rumen smooth muscle

This study investigated the effect of 5 freeze–thaw cycles (freezing at −18°C for 12 h and then thawing at 4°C for approximately 12 h) on the meat quality, proximate composition, water distribution and microstructure of bovine rumen smooth muscle (BSM). As the number of freeze–thaw cycles increased, BSM pH, shear force, water content and protein content decreased by 3.06%, 35.50%, 14.49% and 21.11%, respectively, whereas BSM thawing loss, cooking loss, pressing loss, total aerobic count (TAC), ash content and fat content increased by 108.12%, 47.75%, 78.33%, 90.99%, 105% and 35.20%, respectively. The freeze–thaw cycles resulted in greater protein and lipid oxidation, as evidenced by a 36.46% reduction in the sulfhydryl content and a 209.06% and 338.46% increase in the carbonyl and malondialdehyde contents, respectively. Ice crystal formation disrupted the structural integrity of the muscle tissue. Low-field nuclear magnetic resonance results showed that the freeze–thaw cycles prolonged the relaxation times (T_2b, T₂₁ and T₂₂), indicating that immobile water shifted to free water, and consequently, free water mobility increased. After 3 freeze–thaw cycles, the decline in shear force slowed, the increase in thawing loss became accelerated, and the TAC approached the domain value (6 log colony-forming units/g). Therefore, the number of freeze–thaw cycles of smooth muscle during transport, storage and distribution should be controlled to 3 or fewer. The current results provide a theoretical basis and data support for the further utilisation and culinary processing of smooth muscle.     

南美白对虾肝胰腺蛋白提取方法优化及无水保活环境胁迫对细胞凋亡蛋白的影响

为提高Western Blotting结果的可靠性，以蛋白溶出率和凝胶电泳蛋白条带完整性为指标，比较提取液种类、研磨方式、酶抑制剂种类及其体积分数和提取时间对南美白对虾肝胰腺蛋白提取效果的影响。采用优化后的提取方法获得高质量蛋白样品，并采用Western Blotting法分析无水环境胁迫后南美白对虾肝胰腺组织中细胞凋亡信号通路相关蛋白的表达水平。结果表明：RIPA裂解液作为提取溶剂所得的蛋白溶出率高于水提和磷酸盐缓冲液，电动匀浆和液氮研磨所得蛋白条带更完整，4%蛋白酶磷酸酶混合抑制剂能有效抑制肝胰腺内源酶引起的蛋白降解；采用Western Blotting法分析无水保活期间南美白对虾肝胰腺蛋白，发现低温诱导休眠的同时会引起细胞轻微凋亡，且凋亡水平呈应激时间依赖性增加，环境胁迫解除后有所回调。     

Assessment of dietary protein supplementation on milk productivity of commercial organic dairy farms during the grazing season

Variability of protein and energy supply from pasture during the grazing season is a primary factor that can influence milk production of grazing organic dairy herds in the Northeast United States. This study evaluated the effects of altering the crude protein (CP) content of dietary supplements included in dairy rations fed to grazing organic dairy herds, on milk production and composition. Six commercial organic farms participated in a 6-wk trial, consisting of a 2-wk baseline period and 4-wk experimental period. Farms were paired by their summer 2017 milk urea nitrogen profile, and farms within each pair were assigned by restricted randomization to (1) continuation of their regular supplements (n = 3, control group, CON), or (2) supplement with altered CP as percentage of dry matter, formulated using an organic barley and roasted soybean mix (n = 3, treatment group, TRT). Throughout the 6-wk trial, individual milk samples were collected at 2 consecutive milkings weekly, while pasture and supplement samples, pasture measurements, and management information were collected twice weekly per farm. Data were statistically analyzed using the MIXED procedure of SAS (version 9.4, SAS Institute Inc.) for all parameters, and effects of treatment, week, and their interaction (treatment × week) were determined. The supplement CP (percentage of dry matter) during the baseline period was 13.5% for CON and 15.3% for TRT and 14.8% for CON and 19.3% for TRT during the experimental period. Milk production was 21% higher during the experimental period for TRT compared with CON (24.1 vs. 19.9 kg of milk per day, respectively). Milk production decreased for CON from wk 1 to wk 6 (23.6 vs. 20.4 kg of milk per day), whereas TRT maintained milked production from wk 1 to wk 6 (22.8 vs. 22.7 kg of milk per day). Milk composition was different between groups, with CON having higher fat percent (4.21 vs. 3.73%, respectively) and protein percent (3.15 vs. 3.05%, respectively) compared with TRT for the 6 wk. The milk urea nitrogen concentrations were similar between TRT and CON for the baseline period (11.9 vs. 12.1 mg/dL) and the final week of the experimental period (14.5 vs. 14.2 mg/dL). Although the effects of different diet CP fractions, particularly rumen undegradable protein and soluble protein, must be further delineated, these results indicate that altering the CP content of dietary supplements fed to grazing organic dairy cattle during the summer period in the Northeast US could be a useful mechanism to maintain milk production.     

Adaptation to Endoplasmic Reticulum Stress Enhances Resistance of Oral Cancer Cells to Cisplatin by Up-Regulating Polymerase η and Increasing DNA Repair Efficiency

Endoplasmic reticulum (ER) stress response is an adaptive program to cope with cellular stress that disturbs the function and homeostasis of ER, which commonly occurs during cancer progression to late stage. Late-stage cancers, mostly requiring chemotherapy, often develop treatment resistance. Chemoresistance has been linked to ER stress response; however, most of the evidence has come from studies that correlate the expression of stress markers with poor prognosis or demonstrate proapoptosis by the knockdown of stress-responsive genes. Since ER stress in cancers usually persists and is essentially not induced by genetic manipulations, we used low doses of ER stress inducers at levels that allowed cell adaptation to occur in order to investigate the effect of stress response on chemoresistance. We found that prolonged tolerable ER stress promotes mesenchymal–epithelial transition, slows cell-cycle progression, and delays the S-phase exit. Consequently, cisplatin-induced apoptosis was significantly decreased in stress-adapted cells, implying their acquisition of cisplatin resistance. Molecularly, we found that proliferating cell nuclear antigen (PCNA) ubiquitination and the expression of polymerase η, the main polymerase responsible for translesion synthesis across cisplatin-DNA damage, were up-regulated in ER stress-adaptive cells, and their enhanced cisplatin resistance was abrogated by the knockout of polymerase η. We also found that a fraction of p53 in stress-adapted cells was translocated to the nucleus, and that these cells exhibited a significant decline in the level of cisplatin-DNA damage. Consistently, we showed that the nuclear p53 coincided with strong positivity of glucose-related protein 78 (GRP78) on immunostaining of clinical biopsies, and the cisplatin-based chemotherapy was less effective for patients with high levels of ER stress. Taken together, this study uncovers that adaptation to ER stress enhances DNA repair and damage tolerance, with which stressed cells gain resistance to chemotherapeutics.