High-Resolution Performance of Polycaprolactone Functionalized with Guanidinium Ionic Liquid for Gas Chromatography

This work firstly reported a new polycaprolactone based material functionalized with guanidinium ionic liquid (PCL-GIL) as the stationary phase with high resolution performance for capillary gas chromatography (GC). It is composed of polycaprolactone (PCL) and guanidinium ionic liquid (GIL) with amphiphilic conformation. The PCL-GIL capillary column coated by static method exhibited high column efficiency of 3942 plates/m and moderate polarity. As a result, the PCL-GIL column exhibited high-resolution capability. For a mixture of 27 analytes with a wide ranging polarity and outperformed the PCL-2OH and HP-35 columns, showing its advantageous separation capability for analytes of diverse types. Moreover, the PCL-GIL column showed high resolving capability for various positional isomers and cis-/trans-isomers, including alkylbenzenes, chlorobenzenes, naphthalenes, bromonitrobenzenes, chloronitrobenzenes, benzaldehydes, phenols, alcohols, respectively. In a word, PCL derivatized by GIL units as a new type of stationary phase has a promising future in GC separations.     

牛蛙皮胶原蛋白酸提取条件优化及其对细胞生长和粘附性的影响

应用羟脯氨酸法测定了牛蛙皮胶原蛋白含量,通过正交实验对牛蛙皮胶原蛋白酸法提取条件进行了优化,并结合MTT法测定了胶原蛋白对细胞生长和粘附性的影响。结果显示,牛蛙皮胶原蛋白的含量约为45．1％;温度条件对提取率影响最大,其次依次为酸种、时间和浓度,最优组合为乙酸、1．5mol／L、37℃、24h;在低浓度时,牛蛙皮胶原蛋白对正常人类肝细胞的生长和粘附性无显著影响,当浓度升高到一定值时,对细胞生长和粘附性有显著促进作用。     

Subcellular Distribution and Chemical Forms of Cadmium in the Medicine Food Homology Plant <i>Platycodon grandiflorum</i> (Jacq.) A.DC.

Although Platycodon grandiflorum (Jacq.) A.DC. is a renowned medicine food homology plant, reports of excessive cadmium (Cd) levels are common, which affects its safety for clinical use and food consumption. To enable its Cd levels to be regulated or reduced, it is necessary to first elucidate the mechanism of Cd uptake and accumulation in the plant, in addition to its detoxification mechanisms. This present study used inductively couple plasma-mass-spectrometry to analyze the subcellular distribution and chemical forms of Cd in different tissues of P. grandiflorum. The experimental results showed that Cd was mainly accumulated in the roots [predominantly in the cell wall (50.96%–61.42%)], and it was found primarily in hypomobile and hypotoxic forms. The proportion of Cd in the soluble fraction increased after Cd exposure, and the proportion of insoluble phosphate Cd and oxalate Cd increased in roots and leaves, with a higher increase in oxalate Cd. Therefore, it is likely that root retention mechanisms, cell wall deposition, vacuole sequestration, and the formation of low mobility and low toxicity forms are tolerance strategies for Cd detoxification used by P. grandiflorum. The results of this study provide a theoretical grounding for the study of Cd accumulation and detoxification mechanisms in P. grandiflorum, and they can be used as a reference for developing Cd limits and standards for other medicine food homology plants.     

Crystal structure of the kinase and UBA domains of SNRK reveals a distinct UBA binding mode in the AMPK family

Sucrose non-fermenting (Snf1)-related kinase (SNRK) is a novel member of the AMP-activated protein kinase (AMPK) family and is involved in many metabolic processes. Here we report the crystal structure of an N-terminal SNRK fragment containing kinase and adjacent ubiquitin-associated (UBA) domains. This structure shows that the UBA domain binds between the N- and C-lobes of the kinase domain. The mode of UBA binding in SNRK largely resembles that in AMPK and brain specific kinase (BRSK), however, unique interactions play vital roles in stabilizing the KD-UBA interface of SNRK. We further propose a potential role of the UBA domain in the regulation of SNRK kinase activity. This study provides new insights into the structural diversities of the AMPK kinase family.     

Engineering of cell membrane to enhance heterologous production of hyaluronic acid in Bacillus subtilis

Hyaluronic acid (HA) is a high‐value biopolymer used in the biomedical, pharmaceutical, cosmetic, and food industries. Current methods of HA production, including extraction from animal sources and streptococcal cultivations, are associated with high costs and health risks. Accordingly, the development of bioprocesses for HA production centered on robust “Generally Recognized as Safe (GRAS)” organisms such as Bacillus subtilis is highly attractive. Here, we report the development of novel strains of B. subtilis in which the membrane cardiolipin (CL) content and distribution has been engineered to enhance the functional expression of heterologously expressed hyaluronan synthase (HAS) of Streptococcus equisimilis (SeHAS), in turn, improving the culture performance for HA production. Elevation of membrane CL levels via overexpressing components involved in the CL biosynthesis pathway, and redistribution of CL along the lateral membrane via repression of the cell division initiator protein FtsZ resulted in increases to the HA titer of up to 204% and peak molecular weight of up to 2.2 MDa. Moreover, removal of phosphatidylethanolamine and neutral glycolipids from the membrane of HA‐producing B. subtilis via inactivation of pssA and ugtP, respectively, has suggested the lipid dependence for functional expression of SeHAS. Our study demonstrates successful application of membrane engineering strategies to develop an effective platform for biomanufacturing of HA with B. subtilis strains expressing Class I streptococcal HAS.     

Loss of rice PARAQUAT TOLERANCE 3 confers enhanced resistance to abiotic stresses and increases grain yield in field

Plants frequently suffer from environmental stresses in nature and have evolved sophisticated and efficient mechanisms to cope with the stresses. To balance between growth and stress response, plants are equipped with efficient means to switch off the activated stress responses when stresses diminish. We previously revealed such an off-switch mechanism conferred by Arabidopsis PARAQUAT TOLERANCE 3 (AtPQT3) encoding an E3 ubiquitin ligase, knockout of which significantly enhances resistance to abiotic stresses. To explore whether the rice homologue OsPQT3 is functionally conserved, we generated three knockout mutants with CRISPR-Cas9 technology. The OsPQT3 knockout mutants (ospqt3) display enhanced resistance to oxidative and salt stress with elevated expression of OsGPX1, OsAPX1 and OsSOD1. More importantly, the ospqt3 mutants show significantly enhanced agronomic performance with higher yield compared with the wild type under salt stress in greenhouse as well as in field conditions. We further showed that OsPQT3 expression rapidly decreased in response to oxidative and other abiotic stresses as AtPQT3 does. Taken together, these results show that OsPQT3 is functionally well conserved in rice as an off-switch in stress response as AtPQT3 in Arabidopsis. Therefore, PQT3 locus provides a promising candidate for crop improvement with enhanced stress resistance by gene editing technology.     

利用免疫抑制小鼠模型检测真菌病毒对黄曲霉菌致病力影响

【目的】构建用于比较黄曲霉（Aspergillus flavus,A. flavus）菌株之间致病力差异的小鼠感染模型,并利用该模型评价真菌病毒AfPV1对宿主A. flavus致病力的影响。【方法】用不同浓度环磷酰胺腹腔注射Institute of Cancer Research （ICR）小鼠,根据白细胞的数量判断小鼠免疫抑制程度;通过滴鼻和尾静脉两种感染方法接种不同浓度的A. flavus孢子量,统计14 d以内小鼠的死亡率,确定A. flavus最佳的孢子接种量;通过小鼠组织的菌落负荷量以及肺部组织的病理观察,确定A. flavus感染是否成功;最后利用该小鼠模型评价真菌病毒AfPV1对寄主A. flavus致病力的影响。【结果】腹腔注射环磷酰胺的浓度为250 mg/kg时,能够达到免疫抑制水平;小鼠组织真菌负荷和病理组织切片观察显示A. flavus成功感染接种的ICR小鼠组织;在滴鼻接种模型中,A. flavus的孢子接种量为40μL（1×10⁶CFU/mL）时比较合适评价A. flavus菌株之间的差异;在尾静脉接种的模型中,A. flavus的孢子...     

动物对孢子植物的传播模式及进化意义

孢子植物物种多样性丰富, 是自然生态系统的重要组成部分。孢子植物的传播通常被认为主要依靠风、水、弹力等非生物媒介, 而动物的作用往往被忽略。本文主要概述了: (1)孢子植物对动物传播的适应: 一方面孢子植物可为动物提供食物、庇护所、繁殖场所等, 另一方面孢子植物也可产生视觉、嗅觉等方面的线索来吸引动物, 从而促进动物传播其繁殖体。(2)动物对孢子植物的传播模式: 包括体内传播(消化道和组织寄生)和体外传播两种, 这些模式都能对孢子植物繁殖体进行有效传播。由于动物间形态或生活习性的不同, 以致传播距离存在差异, 最短距离为0.1 cm, 最长距离可从北半球至南半球。(3)动物对孢子植物传播的生态与进化意义; 由于某些孢子植物繁殖体的结构特点或萌发的需求, 以致其繁殖体只能通过动物的传播才能得以定殖, 因此动物与孢子植物之间存在密不可分的关系。目前, 动物对孢子植物的传播研究主要是描述性的内容以及研究单方面的传播途径, 建议在今后的研究中考虑动物对孢子植物传播的有效性以及多途径同时传播对孢子植物定殖的影响, 同时应更加关注孢子植物和动物互惠关系的形成、维持机制及将来的进化趋势。     

Existence of memory in membrane channels: analysis of ion current through a voltage‐dependent potassium single channel

Ion current fluctuation of voltage‐dependent potassium channel in LβT2 cells has been investigated by autocorrelation function and DFA (detrended fluctuation analysis) methods. The calculation of the autocorrelation function exponent and DFA exponent of the sample was based on the digital signals or the 0–1 series corresponding to closing and opening of channels after routine evolution, rather than the sequence of sojourn times. The persistent character of the correlation of the time series was evident from the slow decay of the autocorrelation function. DFA exponent α was significantly greater than 0.5. The main outcome has been the demonstration of the existence of memory in this ion channel. Thus, the ion channel current fluctuation provided information about the kinetics of the channel protein. The result suggests the correlation character of the ion channel protein non‐linear kinetics indicates whether the channel is open or not.