Effect of density on larval development and female productivity ofTisbe holothuriae (Copepoda,Harpacticoida) under laboratory conditions

The harpacticoid copepodTisbe holothuriae has been cultivated in the Helgoland laboratory for more than 20 years. The effects of density on the larval development and the female productivity were studied by comparing two culture systems: (1) enclosed system, and (2) running-water system. In both systems, a nutritious mixed diet ofDunaliella tertiolecta, Skeletonema costatum, and granulatedMytilus edulis was offered. Larval mortality, larval development and female productivity are found to be significantly dependent on both the population density and specificity of the culture system. Increasing density causes higher larval mortality, longer larval development time, and a reduction in female productivity. In comparison with the enclosed system, the running-water system shows decisive advantages: larval mortality is about 20% lower, the rate of larval development is about two days shorter, and there is a very high rate of nauplii production. The sex ratio exhibits high variations, but in general, there is no clear relationship between sex ratio and population density. Nevertheless, when reared in the running-water system, a relatively high percentage of females (>45%) was found at lower densities.     

Rice ubiquitin-conjugating enzyme OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a

Ubc13 is required for Lys63-linked polyubiquitination and innate immune responses in mammals, but its functions in plant immunity still remain largely unknown. Here, we used molecular biological, pathological, biochemical, and genetic approaches to evaluate the roles of rice OsUbc13 in response to pathogens. The OsUbc13-RNA interference (RNAi) lines with lesion mimic phenotypes displayed a significant increase in the accumulation of flg22- and chitin-induced reactive oxygen species, and in defence-related genes expression or hormones as well as resistance to Magnaporthe oryzae and Xanthomonas oryzae pv oryzae. Strikingly, OsUbc13 directly interacts with OsSnRK1a, which is the α catalytic subunit of SnRK1 (sucrose non-fermenting-1-related protein kinase-1) and acts as a positive regulator of broad-spectrum disease resistance in rice. In the OsUbc13-RNAi plants, although the protein level of OsSnRK1a did not change, its activity and ABA sensitivity were obviously enhanced, and the K63-linked polyubiquitination was weaker than that of wild-type Dongjin (DJ). Overexpression of the deubiquitinase-encoding gene OsOTUB1.1 produced similar effects with inhibition of OsUbc13 in affecting immunity responses, M. oryzae resistance, OsSnRK1a ubiquitination, and OsSnRK1a activity. Furthermore, re-interfering with OsSnRK1a in one OsUbc13-RNAi line (Ri-3) partially restored its M. oryzae resistance to a level between those of Ri-3 and DJ. Our data demonstrate OsUbc13 negatively regulates immunity against pathogens by enhancing the activity of OsSnRK1a.     

谷氨酸棒杆菌代谢工程高效生产L-缬氨酸北大核心CSCD

L-缬氨酸作为一种支链氨基酸,广泛应用于医药和饲料等领域。本研究借助多种代谢工程策略相结合的方法,构建了生产L-缬氨酸的微生物细胞工厂,实现了L-缬氨酸的高效生产。首先,通过增强糖酵解途径、减弱副产物代谢途径相结合的方式,强化了L-缬氨酸合成前体丙酮酸的供给;其次,针对L-缬氨酸合成路径关键酶—乙酰羟酸合酶进行定点突变,提高了菌株的抗反馈抑制能力,并利用启动子工程策略,优化了路径关键酶的基因表达水平;最后,利用辅因子工程策略,改变了乙酰羟酸还原异构酶和支链氨基酸转氨酶的辅因子偏好性,由偏好NADPH转变为偏好NADH,从而提高了L-缬氨酸的合成能力。在5L发酵罐中,最优谷氨酸棒杆菌工程菌株Corynebacterium glutamicum K020的L-缬氨酸产量、得率和生产强度分别达到了110g/L、0.51g/g和2.29 g/(L·h)。     

-cyano-substituted analogoues of decarboxylated S-adenosylmethionine as enzyme activated, irreversible inhibitors of S-adenosylmethionine decarboxylase

A pair of -cyano analogues of decarboxylated S-adenosylmethionine (2a and 2b) were synthesized as potential enzyme activated, irreversible inhibitors of the[pyruvoyl enzyme S-adenosylmethionine decarboxylase (AdoMet-DC). Each of these analogues acts as an irreversible inactivator for ADoMet-DC from Escherichia coli (IC₅₀ values of 9 and 50 μM, respectively). These analogues also inactivate human AdoMet-DC, with K_I values of 246.6 and 7.2 μM, and k_inact values of 0.29 and 0.03 min⁻¹, respectively.     

论Paracarinachitids

研究证明Paracarinachitids的外骨骼都不是由8个骨板组成,骨片具有清楚的微层构造,表明它们是由叠覆的生长层组成,每个生长节上有一个小刺,在顶端下侧面前端都有肾形缺凹。同一标本上存在顶端部的融合和远顶端部的分节彼此递交的现象表明这种表象上的差异是后期次生磷灰石包裹或腐蚀的结果;其下侧缘融合和下侧面无缝隙的现象说明将Paracarinachites spinus的骨片分成头板、中间板和尾板是没有根据的。骨片表面存在的针状体是成岩作用的产物,它是粘土矿物和磷酸盐针状结晶体而不是多板纲的骨针。凡此强有力地说明Yangtzechiton是Paracarinachites的晚出同义名。根据形态功能、组织结构,特别是叠覆状排列的骨片和孤立的生长节等特征,paracarinachitids 可能与zhijinitids有密切关系,但两者是否同源尚缺少进一步的证据、据此,笔者认为将paracarinachitids 暂置于分类位置未定的具骨片后生动物较妥。     

Preparation of the pure diastereomeric forms of S- (5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2- cyclopentene and their evaluation as irreversible inhibitors of S-adenosylmethionine decarboxylase from Escherichia coli

The conformationally restricted S-adenosylmethionine analogue AdoMac (S-(5′-deoxy-5′-adenosyl)-1-ammonio-4-methylsulfonio-2-cyclopentene has been shown to act as an enzyme activated, irreversible inhibitor of theEscherichia coli form of the enzyme S-adenosylmethionine decarboxylase. Inactivation of the enzyme is presumably initiated by formation of an imine linkage between the inhibitor and the terminal pyruvate of the enzyme, followed by base-catalyzed elimination of methylthioadenosine and generation of a latent electrophile. Removal of the driving force for the elimination of methylthioadenosine resulted in a reversibly binding inhibitor. Thus, the thioether analogue corresponding to AdoMac, and the corresponding dihydro derivative (H₂-AdoMac), reversibly inhibit the enzyme. AdoMac was resolved into its four pure diastereomeric forms, and each diastereomer was evaluated as an irreversible inhibitor of the enzyme. The K_I values for the individual diastereomers range between 3.83 and 39.6 μM, with the cis-1S,4R diastereomer being the most potent inhibitor. However, the k_inact values for the four diastereomers are not significantly different, suggesting that the binding of each diastereomer to the enzyme is configuration-dependent, while the subsequent inactivation likely proceeds through a single intermediate which is formed from each of the four diastereomers. Since each pure diastereomer represents a distinct conformational mimic exhibiting restricted sidechain rotation, the data suggests that these and related analogues may be useful as conformational probes for the catalytic site of AdoMet-DC.     

大白菜NHX基因家族的鉴定与表达分析

Na+/H+逆向转运蛋白(Na+/H+antiporter,NHX)基因家族在植物响应盐胁迫中发挥重要作用。本研究鉴定了大白菜NHX基因家族成员,并分析了大白菜NHX基因(Brassica rapa ssp.Pekinensis NHX,BrNHXs)响应高温、低温、干旱和盐胁迫等非生物逆境的表达模式。结果表明,在大白菜中共鉴定到9个NHX基因家族成员,分布在大白菜的6条染色体上,其氨基酸数目在513–1154 aa之间,相对分子量集中在56804.22–127856.66 kDa,等电点位于5.35–7.68之间。该基因家族成员主要存在于液泡中,基因结构完整,外显子的数目介于11–22之间。大白菜NHX基因家族编码的蛋白质二级结构都具有α-螺旋、β-转角和不规则卷曲结构,其中α-螺旋发生频率较高。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)分析显示,该基因家族成员在高温、低温、干旱和盐胁迫下均有不同程度地响应,且在不同时间表达差异显著。以BrNHX02和BrNHX09对这4种胁迫的响应最为显著,表达量在处理72 h时均显著上调,可作为候选基因进一步验证其功能。     

Nonparametric and semiparametric estimation with sequentially truncated survival data

In observational cohort studies with complex sampling schemes, truncation arises when the time to event of interest is observed only when it falls below or exceeds another random time, that is, the truncation time. In more complex settings, observation may require a particular ordering of event times; we refer to this as sequential truncation. Estimators of the event time distribution have been developed for simple left-truncated or right-truncated data. However, these estimators may be inconsistent under sequential truncation. We propose nonparametric and semiparametric maximum likelihood estimators for the distribution of the event time of interest in the presence of sequential truncation, under two truncation models. We show the equivalence of an inverse probability weighted estimator and a product limit estimator under one of these models. We study the large sample properties of the proposed estimators and derive their asymptotic variance estimators. We evaluate the proposed methods through simulation studies and apply the methods to an Alzheimer's disease study. We have developed an R package, seqTrun , for implementation of our method.     

LEP and LEPR are possibly a double-edged sword for wound healing

Wound healing is a complex and error-prone process. Wound healing in adults often leads to the formation of scars, a type of fibrotic tissue that lacks skin appendages. Hypertrophic scars and keloids can also form when the wound-healing process goes wrong. Leptin (Lep) and leptin receptors (LepRs) have recently been shown to affect multiple stages of wound healing. This effect, however, is paradoxical for scarless wound healing. On the one hand, Lep exerts pro-inflammatory and profibrotic effects; on the other hand, Lep can regulate hair follicle growth. This paper summarises the role of Lep and LepRs on cells in different stages of wound healing, briefly introduces the process of wound healing and Lep and LepRs, and examines the possibility of promoting scarless wound healing through spatiotemporal, systemic, and local regulation of Lep levels and the binding of Lep and LepRs.