着-聚赖氨酸/淀粉抑菌膜的制备及性能检测

对以淀粉为成膜基质,海藻酸钠和甘油为增塑剂,ε-聚赖氨酸为抑菌剂的淀粉基抑菌膜进行研究,并对其抑菌性和膜性能进行检测。结果表明,ε-聚赖氨酸对大肠杆菌、枯草芽孢杆菌和酵母菌的最低抑菌浓度分别为1、2、2μg/mL,而将ε-聚赖氨酸添加到淀粉膜后,最低抑菌浓度都提高到64 mg/g,对淀粉膜和抑菌膜的力学性能、红外光谱检测、透气性和透油性及膜液的流变性、接触角等比较,差异较小。     

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.     

食品中2株非常见耶尔森菌分离鉴定及耐药性分析

目的 对食品中两株Yersinia aleksiciae(Ya)进行分离鉴定及耐药性分析研究,为非常见耶尔森菌属菌株的分离鉴定提供技术储备。方法 2018年8月至2019年3月采集猪肉、牛肉、鸡肉食品样品,共300份。对分离到的疑似Ya菌株完成生化鉴定、耐药表型鉴定和全基因组测序分析。结果 食品样品中Ya的检出率为0.7%(2/300),生化鉴定中Ya与小肠结肠炎耶尔森菌(Yersinia enterocolitica,Ye)最大区别是Ye为赖氨酸脱羧酶阴性,Ya为赖氨酸脱羧酶阳性。Ya暂未发现与Ye类似的对氨苄西林、I代头孢菌素、替卡西林等抗生素的天然耐药性,两株Ya菌株的耐药性不同。全基因组测序比对16S rRNA和gyrB、rpoD基因三个片段序列,证实两株分离菌株为Ya。结论 分离到的两株Ya为国内首次发现,目前国外鲜有从食品中分离出该菌的报道。本研究提供的方法可用于其他非常见耶尔森菌的分离鉴定。     

Influence of ammonia and lysine supplementation on yeast growth and fermentation with respect to gluten-free type brewing using unmalted sorghum grain

Because gluten-free type brewing with unmalted sorghum does not provide adequate nitrogen for complete fermentation, wort supplementation with ammonia (as diammonium phosphate, DAP) or lysine on yeast performance was investigated. By Phenotype Microarray, under aerobic conditions, greater yeast growth was indicated with DAP than lysine both as a single source and combined with sorghum wort amino acids. With sorghum fermentation, both DAP and lysine improved maltose and maltotriose uptake. However, DAP supplementation also maintained yeast numbers (24.0–21.3 × 10⁶ cells mL⁻¹), whereas there was a decline with lysine supplementation. Lysine supplementation also resulted in adverse effects on yeast cell morphology. Neither DAP nor lysine supplementation resulted in evident genetic change to the yeast, but the change in substrate from barley malt wort to unmalted sorghum wort slightly altered the yeast genetically. Therefore, ammonia as DAP has potential as a nitrogen supplement for improving yeast fermentation performance in sorghum gluten-free brewing.     

Design,Synthesis, and Biological Evaluation of Lysine Demethylase 5 C Degraders

Lysine demethylase 5 C (KDM5C) controls epigenetic gene expression and is attracting great interest in the field of chemical epigenetics. KDM5C has emerged as a therapeutic target for anti-prostate cancer agents, and recently we identified triazole 1 as an inhibitor of KDM5C. Compound 1 exhibited highly potent KDM5C-inhibitory activity in in vitro enzyme assays, but did not show strong anticancer effects. Therefore, a different approach is needed for the development of anticancer agents targeting KDM5C. Here, we attempted to identify KDM5C degraders by focusing on a protein-knockdown strategy. Compound 3 b , which was designed based on compound 1 , degraded KDM5C and inhibited the growth of prostate cancer PC-3 cells more strongly than compound 1 . These findings suggest that KDM5C degraders are more effective as anticancer agents than compounds that only inhibit the catalytic activity of KDM5C.     

Effects of graded removal of lysine from an intravenously infused amino acid mixture on lactation performance and mammary amino acid metabolism in lactating goats

To investigate responses of milk protein synthesis and mammary AA metabolism to a graded decrease of postruminal Lys supply, 4 lactating goats fitted with jugular vein, mammary vein, and carotid artery catheters and transonic blood flow detectors on the external pudic artery were used in a 4 × 4 Latin square experiment. Goats were fasted for 24 h and then received a 9-h intravenous infusion of an AA mixture plus glucose. Milk yield was recorded and samples were taken in h 2 to 8 of the infusion period; a mammary biopsy was performed in the last hour. Treatments were graded decrease of lysine content in the infusate to 100 (complete), 60, 30, or 0% as in casein. Lysine-removed infusions linearly decreased milk yield, tended to decrease lactose yield, and tended to increase milk fat to protein ratio. Milk protein content and yield were linearly decreased by graded Lys deficiency. Mammary Lys uptake was concomitantly decreased, but linear regression analysis found no significant relationship between mammary Lys uptake and milk protein yield. Treatments had no effects on phosphorylation levels of the downstream proteins measured in the mammalian target or rapamycin pathway except for a tended quadratic effect on that of eukaryotic initiation factor 2, which was increased and then decreased by graded Lys deficiency. Removal of Lys from the infusate linearly increased circulating glucagon and glucose. Removal of Lys from the infusate linearly decreased arterial and venous concentrations of Lys. Treatments also had a significant quadratic effect on venous Lys, suggesting mechanisms to stabilize circulating Lys at a certain range. The 2 infusions partially removing Lys resulted in a similar 20% decrease, whereas the 0% Lys infusion resulted in an abrupt 70% decrease in mammary Lys uptake compared with that of the full-AA mixture infusion. Consistent with the abrupt decrease, mammary Lys uptake-to-output ratio decreased from 2.2 to 0.92, suggesting catabolism of Lys in the mammary gland could be completely prevented when the animal faced severe Lys deficiency. Mammary blood flow was linearly increased, consistent with the linearly increased circulating nitric oxide by graded Lys deficiency, indicating mechanisms to ensure the priority of the mammary gland in acquiring AA for milk protein synthesis. Infusions with Lys removed increased mammary clearance rate of Lys numerically by 2 to 3 fold. In conclusion, the decreased milk protein yield by graded Lys deficiency was mainly a result of the varied physiological status, as indicated by the elevated circulating glucagon and glucose, rather than a result of the decreased mammary Lys uptake or depressed signals in the mTOR pathway. Mechanisms of Lys deficiency to promote glucagon secretion and mammary blood flow and glucagon to depress milk protein synthesis need to be clarified by future studies.     

Quantitative Proteomic Approach Reveals Altered Metabolic Pathways in Response to the Inhibition of Lysine Deacetylases in A549 Cells under Normoxia and Hypoxia

Growing evidence is showing that acetylation plays an essential role in cancer, but studies on the impact of KDAC inhibition (KDACi) on the metabolic profile are still in their infancy. Here, we analyzed, by using an iTRAQ-based quantitative proteomics approach, the changes in the proteome of KRAS-mutated non-small cell lung cancer (NSCLC) A549 cells in response to trichostatin-A (TSA) and nicotinamide (NAM) under normoxia and hypoxia. Part of this response was further validated by molecular and biochemical analyses and correlated with the proliferation rates, apoptotic cell death, and activation of ROS scavenging mechanisms in opposition to the ROS production. Despite the differences among the KDAC inhibitors, up-regulation of glycolysis, TCA cycle, oxidative phosphorylation and fatty acid synthesis emerged as a common metabolic response underlying KDACi. We also observed that some of the KDACi effects at metabolic levels are enhanced under hypoxia. Furthermore, we used a drug repositioning machine learning approach to list candidate metabolic therapeutic agents for KRAS mutated NSCLC. Together, these results allow us to better understand the metabolic regulations underlying KDACi in NSCLC, taking into account the microenvironment of tumors related to hypoxia, and bring new insights for the future rational design of new therapies.     

Natural polysaccharides platforms for oral controlled release of ketoprofen lysine salt

Context: Ketoprofen lysinate (KL) is one of the most widely used non-steroidal anti-inflammatory drugs in the symptomatic treatment of some chronic inflammatory diseases. Compared to ketoprofen, KL shows better pharmacokinetics and tolerability. However, due to its short half-life of 1–2?h, a multiple dose regimen is required for oral administration. Thus, the present work deals with its encapsulation in a hydrogel-based system by prilling in order to prolong its activity.

Objective: In this paper, we propose alginate and pectin as carriers and release tailoring agent for the development of hydrogel-based beads for KL retarded and sustained release.

Materials and methods: Beads were produced by a Nisco Encapsulator® using alginate or pectin. Operative variables were optimized to produce beads with desired morphology and size. Solid state properties were analyzed by SEM and DSC. Drug release performance was studied by Pharmacopeia pH-change assay to simulate gastrointestinal environment.

Results and discussion: Prilling technique was successfully used to encapsulate high soluble drugs as KL in polysaccharides-based hydrogels. Pectin proved to be a proper polymer able to encapsulate ketoprofen lysine salt. Formulation (F8) showed good morphological properties and size, high drug content (15.6%) and encapsulation efficiency (93.5%) and promising drug release profiles. Hosting F8 in an acid-resistant capsule (DR^®caps) a delivery platform has been developed to control KL release in a delayed (90?min lag time) and prolonged way (270?min complete release).

Conclusion: The platform may be proposed as potentially useful in the oral administration of NSAIDs in chronic inflammatory diseases affected by circadian rhythm.     

Possible adducts formed between hydroxymethylfurfural and selected amino acids,and their release in simulated gastric model

This work aimed to investigate the reactivity of hydroxymethylfurfural (HMF) with selected amino acids, to identify the produced adducts and to clarify whether or not the adducts release HMF after their digestion under gastric conditions. Results showed that cysteine, glycine and lysine can deplete the added HMF, and their reactivity increased with increasing pH and temperature. Cysteine (25 μmol mL^?1) depleted 91.0% of the added HMF (315.3 μg mL^?1) at 40 °C in 15 min, lysine did not eliminate HMF until 80 °C, and glycine started to eliminate HMF at 100 °C. Four adducts for cysteine, three adducts for lysine and only one adduct for glycine were identified through HPLC–MS–MS after they reacted with HMF. The adducts formed from the reaction mixture of cysteine, lysine and glycine with HMF only released 1.7%, 2.6% and 10.5% of eliminated HMF, respectively, after their digestion in simulated gastric conditions.     

Effects of dietary rumen‐protected lysine on milk yield and composition in lactating cows fed diets containing double‐low rapeseed meal

This study investigated the effects of dietary rumen‐protected lysine (RPLys) on milk yield and composition in lactating cows fed diets containing double‐low rapeseed meal. Twenty‐four cows were divided into three groups and fed one of three dietary treatments: 10% soya bean meal (SBM) diet (control), 15% double‐low rapeseed meal (DLRSM) diet and 15% DLRSM + 32 g/d RPLys (DLRSML) diet. The results showed that no differences (P > 0.05) were observed in milk yield, 4% fat correction milk (FCM), energy correction milk (ECM), protein yield, milk fat yield, milk fat, lactose yield and lactose between control and DLRSM. Supplementation with DLRSML increased (P < 0.05) milk yield, 4% FCM, ECM, and protein yield compared with the SBM. The results indicated that DLRSM and DLRSML might be used to substitute for SBM as a protein source in lactating cows, and the latter might be more beneficial to improve the performance.