Surface active adrenoceptor compounds prevent the settlement of cyprid larvae of Balanus improvisus

The barnacle Balanus improvisus is the major fouling macroorganism in Swedish waters and it colonizes most man‐made surfaces submerged in the sea. New or impending legislation restricts the use of traditional, hazardous antifouling coatings based on heavy metals, mainly copper and tin. This calls for the development of new non‐toxic methods that prevent barnacle settlement. In this work several adrenoceptor compounds are shown to be very efficient in preventing the settlement of cyprid larvae of B. improvisus. The settlement rate of laboratory‐reared cyprids was studied in hydrophilised polystyrene dishes containing adrenoceptor antagonists and agonists dissolved in seawater. Two of these drugs, medetomidine and clonidine, repeatedly inhibited settlement at concentrations between 1 nM and 10 nM. In the vertebrate adrenoceptor classification system, which separates pharmacological substances according to their receptor affinity, both of these substances are classified as α₂ adrenoceptor agonists. An inhibiting effect on presyn‐aptic receptors is suggested, but the localization of the receptor effect requires futher studies. Experiments also revealed that the inhibiting effect of medetomidine was reversible. Cyprids incubated with medetomidine for 20 h attached and metamorphosed into juvenile barnacles after washing and transferrence to seawater. The antagonizing compound atipamezole reversed the effect of medetomidine. This observation supports the assumption that this substance acts at the receptor level. Studies of the surface affinity of medetomidine revealed a strong tendency to accumulate in solid/ liquid phase boundaries. This ability makes it particularly attractive as a candidate for the development of a slow‐release carrier in marine coatings. Panels coated with medetomidine in an acrylate polymer and exposed in the field reduced the recruitment of B. improvisus by 96% after 4 weeks and by 70% after 8 weeks.     

基因捕捉及其在植物基因分离和功能基因组学上的应用

基因捕捉是一种报告基因的随机整合技术。基因捕捉系统已成为分离基因、鉴定基因功能的重要手段。基因捕捉(gene traps)包括增强子捕捉(enhancer trap)、启动子捕捉(promoter trap)和基因捕捉(gene trap),通称为基因捕捉(gcne traps)。在增强子捕捉中,报告基因与一个基本启动子融合,这个启动子不能使报告基因表达,但可被临近的增强子激活。在启动子捕捉和基因捕捉中,报告基因的启动子被去除,融合基因只有以正确的方向插入到转录单元内才能表达。对基因捕捉系统的结构特征、构建方法、应用范围、研究现状和应用前景等作了系统论述,并对有关问题进行了讨论。     

絮凝颗粒粒度分布对自絮凝酵母SPSC01乙醇耐受能力的影响

利用激光聚焦反射式颗粒测量系统, 通过调节不同的搅拌速率, 得到了分批补料培养条件下粒度分布不同的四个絮凝酵母SPSC01颗粒群体, 进而对絮凝颗粒群体分布对乙醇耐受性进行了系统研究。经过6 h、20%乙醇的冲击, 颗粒粒度为100、200、300和400 mm的自絮凝酵母SPSC01的存活率分别为3.5%、26.7%、48.8%和37.6%。这表明不同粒度分布的絮凝颗粒群体乙醇耐受性具有明显差别, 在一定粒度范围内乙醇耐受性达到最高, 乙醇耐受性最高的酵母群体的乙醇得率系数85.5%, 比乙醇耐性最低的颗粒群体提高了7.2%。粒度为100、200和300 mm的自絮凝酵母颗粒群体总麦角固醇、游离麦角固醇及海藻糖含量与粒度大小成正相关, 但在粒度为400 mm的絮凝颗粒群体中总麦角固醇、游离麦角固醇及海藻糖含量呈下降趋势, 与其乙醇耐性低于300 mm絮凝颗粒的结果相一致。对细胞膜透性的研究表明, 颗粒粒度为300 mm的絮凝酵母颗粒细胞膜通透性(P′)最低, 分别仅为颗粒粒度为100 mm和200 mm颗粒群体的43%和52%, 表明粒度分布不同的絮凝颗粒群体乙醇耐性的差别与细胞膜透性密切相关。     

Ecological and evolutionary patterns of freshwater maturation in Pacific and Atlantic salmonines

Reproductive tactics and migratory strategies in Pacific and Atlantic salmonines are inextricably linked through the effects of migration (or lack thereof) on age and size at maturity. In this review, we focus on the ecological and evolutionary patterns of freshwater maturation in salmonines, a key process resulting in the diversification of their life histories. We demonstrate that the energetics of maturation and reproduction provides a unifying theme for understanding both the proximate and ultimate causes of variation in reproductive schedules among species, populations, and the sexes. We use probabilistic maturation reaction norms to illustrate how variation in individual condition, in terms of body size, growth rate, and lipid storage, influences the timing of maturation. This useful framework integrates both genetic and environmental contributions to conditional strategies for maturation and, in doing so, demonstrates how flexible life histories can be both heritable and subject to strong environmental influences. We review evidence that the propensity for freshwater maturation in partially anadromous species is predictable across environmental gradients at geographic and local spatial scales. We note that growth is commonly associated with the propensity for freshwater maturation, but that life-history responses to changes in growth caused by temperature may be strikingly different than changes caused by differences in food availability. We conclude by exploring how contemporary management actions can constrain or promote the diversity of maturation phenotypes in Pacific and Atlantic salmonines and caution against underestimating the role of freshwater maturing forms in maintaining the resiliency of these iconic species.     

小蓬NeMT2基因的克隆及其植物表达载体的构建

该研究利用RACE ( Rapid amplification of cDNA ends)技术从小蓬中成功分离编码金属硫蛋白( Metal-lothionein,MT)的cDNA序列,命名为NeMT2,在GenBank中登录号为KT835290。该基因全长590 bp,开放阅读框为237 bp,编码78个氨基酸,编码的氨基酸序列中含有14个半胱氨酸残基( Cys,C),呈C-C,C-X-C,C-X-X-C排列,集中分布在肽链的N端和C端,基因编码蛋白的分子量为7.6036 kD,等电点为4.71。系统发育分析表明,小蓬金属硫蛋白NeMT2与藜科的海蓬子( AEF01492)和盐穗木( AHI62953)同源性最高,其次是甜菜( XP 010667708.1)。生物信息学分析表明,金属硫蛋白NeMT2无信号肽结构,属于非跨膜亲水性蛋白;疏水性分析表明,NeMT2蛋白的35~45个氨基酸之间有较强的疏水性,其中第41位Asp具最强的疏水性(1.444);结构预测分析该蛋白质二级结构的主要元件是无规则卷曲。通过RT-PCR对NeMT2基因的表达分析发现, NeMT2基因在铜矿区和非铜矿区的小蓬叶片中均有表达,但该基因在铜矿区小蓬叶片的表达量明显高于非铜矿区。将小蓬NeMT2基因定向克隆到植物表达载体pCAMBIA1300的35S 启动子下游,构建该基因的植物超表达载体pCAMBIA1300＋NeMT2。该研究结果为进一步研究该基因的功能和小蓬响应重金属胁迫的分子机制提供了一定基础。     

Impairment of cytotype regulation of P-element activity in Drosophila melanogaster by mutations in the Su(var)205 gene

Cytotype regulation of transposable P elements in the germ line of Drosophila melanogaster is associated with maternal transmission of P elements inserted at the left telomere of the X chromosome. This regulation is impaired in long-term stocks heterozygous for mutations in Suppressor of variegation 205 [Su(var)205], a gene implicated in the control of telomere length. Regulation by TP5, a structurally incomplete P element at the X telomere, is more profoundly impaired than regulation by TP6, a different incomplete P element inserted at the same site in a TAS repeat at the X telomere. Genetic analysis with the TP5 element indicates that its regulatory ability is not impaired in flies whose fathers came directly from a stock heterozygous for a Su(var)205 mutation, even when the flies themselves carry this mutation. However, it is impaired in flies whose grandfathers came from such a stock. Furthermore, this impairment occurs even when the Su(var)205 mutation is not present in the flies themselves or in their mothers. The impaired regulatory ability of TP5 persists for at least several generations after TP5 X chromosomes extracted from a long-term mutant Su(var)205 stock are made homozygous in the absence of the Su(var)205 mutation. Impairment of TP5-mediated regulation is therefore not directly dependent on the Su(var)205 mutation. However, it is characteristic of the six mutant Su(var)205 stocks that were tested and may be related to the elongated telomeres that develop in these stocks. Impairment of regulation by TP5 is also seen in a stock derived from Gaiano, a wild-type strain that has elongated telomeres due to a dominant mutation in the Telomere elongation (Tel) gene. Regulation by TP6 is not impaired in the Gaiano genetic background. The regulatory abilities of the TP5 and TP6 elements are therefore not equally susceptible to the effects of elongated telomeres in the mutant Su(var)205 and Gaiano stocks.     

Isolation of early hematopoietic cells, including megakaryocyte progenitors, in the ALDH-bright cell population of cryopreserved, banked UC blood

BACKGROUND: ALDH-bright (ALDH(br)) cell populations sorted from freshly collected umbilical cord blood (UCB) on the basis of their high aldehyde dehydrogenase (ALDH) activity are highly enriched for HPC. HPC with low ALDH activity (ALDH(dim)) are primarily short-term progenitors, whereas progenitors that initiate long-term cultures or establish long-term grafts in xenograft models are ALDH(br). We examined the multilineage hematopoietic and platelet progenitor activities of ALDH(br) cells recovered from cryopreserved UCB units typically employed in the practice of clinical transplantation. METHODS: Frozen UCB units were thawed, washed, immunomagnetically depleted of cells expressing glycophorin A and CD14, reacted for flow cytometric detection of ALDH, and sorted to yield ALDH(br) and ALDH(dim) populations. We measured surface Ag expression and viability of cells in the ALDH(br) and ALDH(dim) populations by flow cytometry and hematopoietic (CFC-H) and megakaryocytic (CFC-Mk) colony-forming cells in each population. RESULTS: ALDH(br) populations isolated from thawed UCB cells were highly enriched for CD34(+) and CD133(+) cells. Flow-sorted ALDH(br) populations were enriched 1116-fold in CFC-H, 10-fold in multilineage GEMM colonies and 2015-fold in CFC-Mk compared with the ALDH(dim) population. All progenitors giving rise to large Mk colonies were derived from ALDH(br) populations. DISCUSSION: ALDH(br) populations recovered from thawed, banked UCB with the method we describe have HPC activity and may be useful in the clinic to facilitate reconstitution of erythroid, myeloid and megakaryocytic blood elements.     

三角鲂和长春鳊肌肉营养成分分析与品质评价

用常规方法测定、分析了三角鲂(Megalobrama tarminalis)和长春鳊(Parabramis pekinensis肌肉中营养成分组成与含量.结果显示,三角鲂肌肉蛋白质、脂肪含量分别为18.19％和3.06％,长春鳊肌肉蛋白质、脂肪含量分别为19.38％和2.89％.三角鲂和长春鳊肌肉中均检测出18种氨基酸,其中包括了8种人体必需氨基酸.三角鲂肌肉中氨基酸总量为76.27％,其中,8种人体必需氨基酸含量为32.17％,占氨基酸总量的42.18％;长春鳊肌肉中氨基酸总量为77.60％,其中,8种人体必需氨基酸含量为31.70％,占氨基酸总量的40.85％.必需氨基酸的构成比例基本符合FAO/WHO的标准.三角鲂肌肉中限制性氨基酸主要为甲硫氨酸加胱氨酸,必需氨基酸指数为63.55,4种呈味氨基酸为氨基酸总量的32.81％;长春鳊肌肉中限制性氨基酸主要为色氨酸,必需氨基酸指数为66.81,4种呈味氨基酸为氨基酸总量的33.80％.脂肪酸中二十碳五烯酸(EPA)与二十二碳六烯酸(DHA)含量均较高,三角鲂为7.96％,长春鳊为3.11％.矿物元素比值合理.以上分析表明,三角鲂和长春鳊均为营养价值、经济价值都较高的优质鱼类,相比而言,三角鲂肌肉脂肪、脂肪酸含量和质量更优,而长春鳊肌肉在蛋白质、氨基酸组成与含量方面更优.     

A photoactive phosphonamide derivative of GTP for the identification of the GTP-binding domain in beta-tubulin

A GTP photoaffinity probe (125I-APTG) was developed that incorporated an [125I]-N-(4-azidophenyl)-2-amino-3-(4-hydroxy-3-iodophenyl)propionamide group at the gamma-position of GTP through a phosphonamide linkage. A combination of saturation and GTP protection studies (90% protection at 25 microM GTP with an apparent Kd of 5 microM) validated the use of this new probe as a satisfactory GTP mimic. This probe offered the advantage of possessing an 125I radiolabel external to the GTP moiety, in contrast to the previously reported [gamma 32P]-8-N3GTP that possessed an internal 32P radiolabel. This novel feature accommodated the purification of photolabeled peptides using a combination of ion-exclusion, gel filtration, and HPLC techniques. [125I]APTG was used to identify a peptide (beta:65-79) in the exchangeable GTP-binding domain of the beta-subunit of tubulin.     

冷藏对美洲斑潜蝇蛹期发育和生存的影响

【目的】美洲斑潜蝇Liriomyza sativae Blanchard自20世纪90年代入侵我国后已经成为蔬菜和观赏植物的重要害虫。然而,近些年该昆虫已经逐渐成为研究植物和昆虫、昆虫抗寒性和寄主适应性等热点课题的模式种类。因此,为了实验室条件下更好保藏该虫的种群,本文研究了冷藏(世代发育起点温度8℃)对该斑潜蝇蛹发育和生存的影响。【方法】以化蛹后第3天的蛹为实验对象,8℃低温冷藏,并以冷藏时间(0、7、14、21、28和35 d)为参试因子研究了冷藏时间对美洲斑潜蝇蛹羽化率的影响;化蛹后每24 h解剖观察一次蛹并对蛹期发育阶段进行划分;根据划分的蛹期阶段,对处理中未羽化的蛹进行解剖比对,得到对低温最敏感的蛹期。【结果】随着冷藏时间的增长,美洲斑潜蝇的蛹羽化率逐渐降低。超过14 d后,羽化率明显下降。冷藏至35 d时羽化率为零。解剖正常发育的美洲斑潜蝇的蛹,将蛹期分为预蛹期、隐头蛹期、显头蛹初期、半透明眼期、浅黄色到琥珀色眼期、红褐色眼期、鬃毛蛹期。不同冷藏时间条件下,未羽化的蛹死亡时间主要为鬃毛蛹期,且鬃毛蛹期死亡的蛹的数量与贮藏时间呈正比例关系。【结论】在室内冷藏美洲斑潜蝇时,3日龄蛹8℃贮藏少于14 d为宜;鬃毛蛹期(10~11日龄蛹)对低温比较敏感,冷藏过程中,应避免鬃毛蛹期的蛹。