Signaling pathway of a photoactivable Rac1-GTPase in the early stages

In modern life‐ and medical‐sciences major efforts are currently concentrated on creating artificial photoenzymes, consisting of light‐ oxygen‐voltage‐sensitive (LOV) domains fused to a target enzyme. Such protein constructs possess great potential for controlling the cell metabolism as well as gene function upon light stimulus. This has recently been impressively demonstrated by designing a novel artificial fusion protein, connecting the AsLOV2‐Jα‐photosensor from Avena sativa with the Rac1‐GTPase (AsLOV2‐Jα‐Rac1), and by using it, to control the motility of cancer cells from the HeLa‐line. Although tremendous progress has been achieved on the generation of such protein constructs, a detailed understanding of their signaling pathway after photoexcitation is still in its infancy. Here, we show through computer simulations of the AsLOV2‐Jα‐Rac1‐photoenzyme that the early processes after formation of the Cys450‐FMN‐adduct involve the breakage of a H‐bond between the carbonyl oxygen FMN‐C4?O and the amino group of Gln513, followed by a rotational reorientation of its sidechain. This initial event is followed by successive events including β‐sheet tightening and transmission of torsional stress along the Iβ‐sheet, which leads to the disruption of the Jα‐helix from the N‐terminal end. Finally, this process triggers the detachment of the AsLOV2‐Jα‐photosensor from the Rac1‐GTPase, ultimately enabling the activation of Rac1 via binding of the effector protein PAK1. Proteins 2012; © 2012 Wiley Periodicals, Inc.     

毒蛇咬伤患者院前早期程序化急救处理方案的应用观察

目的探讨毒蛇咬伤患者院前早期程序化急救处理方案的应用价值。方法回顾性分析我院58例毒蛇咬伤患者应用院前早期程序化急救处理方案的临床资料。结果 58例毒蛇咬伤患者经院前早期程序化急救处理后,痊愈出院53例,好转3例,自动出院1例,院前死亡2例（眼镜王蛇咬伤1例,银环蛇咬伤1例）。结论院前急救是蛇伤治疗前期的一个关键性步骤,院前早期程序化急救处理方案应用于蛇伤患者,有利于提高抢救成功率。     

山东海域赤潮灾害特征及预警报管理

掌握赤潮灾害时空分布规律是科学设计赤潮监控方案、提高防灾管理效率的基础和依据。本文统计分析了1990—2010年间山东海域的76次赤潮灾害事件数据。结果表明:赤潮灾害频率显著地分为4个波段,且峰值呈递增趋势;灾害面积呈现周期性倒U型曲线变化;每年的5—10月是赤潮灾害的多发期,其中,8月的灾害次数和灾害面积均占全年总数的30%以上;灾害空间分布集中在莱州湾海域、青岛近海、庙岛群岛的北隍城海域,烟台四十里湾的赤潮灾害最频繁;夜光藻(Noctiluca scintillans)和球型棕囊藻(Phaeocystis globosa)引发的赤潮灾害面积最高,夜光藻和红色裸甲藻(Gymnodinium sanguineum)致灾次数最多。本文从赤潮监测与信息采集、赤潮预报方法以及赤潮灾害预警报管理机制3个方面分析总结了山东海域赤潮灾害预警报业务化管理现状,结合山东海域赤潮灾害特征和预警报管理实践,提出了以控制污染和修复生态环境为根本的赤潮防控建议。     

Exogenous selection shapes germination behaviour and seedling traits of populations at different altitudes in a Senecio hybrid zone

Background and Aims

The Senecio hybrid zone on Mt Etna, Sicily, is characterized by steep altitudinal clines in quantitative traits and genetic variation. Such clines are thought to be maintained by a combination of ‘endogenous’ selection arising from genetic incompatibilities and environment-dependent ‘exogenous’ selection leading to local adaptation. Here, the hypothesis was tested that local adaptation to the altitudinal temperature gradient contributes to maintaining divergence between the parental species, S. chrysanthemifolius and S. aethnensis.

Methods

Intra- and inter-population crosses were performed between five populations from across the hybrid zone and the germination and early seedling growth of the progeny were assessed.

Key Results

Seedlings from higher-altitude populations germinated better under low temperatures (9–13 °C) than those from lower altitude populations. Seedlings from higher-altitude populations had lower survival rates under warm conditions (25/15 °C) than those from lower altitude populations, but also attained greater biomass. There was no altitudinal variation in growth or survival under cold conditions (15/5 °C). Population-level plasticity increased with altitude. Germination, growth and survival of natural hybrids and experimentally generated F₁s generally exceeded the worse-performing parent.

Conclusions

Limited evidence was found for endogenous selection against hybrids but relatively clear evidence was found for divergence in seed and seedling traits, which is probably adaptive. The combination of low-temperature germination and faster growth in warm conditions might enable high-altitude S. aethnensis to maximize its growth during a shorter growing season, while the slower growth of S. chrysanthemifolius may be an adaptation to drought stress at low altitudes. This study indicates that temperature gradients are likely to be an important environmental factor generating and maintaining adaptive divergence across the Senecio hybrid zone on Mt Etna.     

A robust flow-cytometric protocol for assessing growth rate of hatchery-reared barramundi Lates calcarifer larvae

In this study, a flow-cytometric cell cycle analysis method to assess instantaneous growth rate of whole larvae of the Australian barramundi Lates calcarifer was developed and validated. High-resolution DNA measurements of either fresh, frozen or RNAlater-preserved larvae (gap0-gap1, G(0) -G(1), coefficient of variation (c.v.) < 3, 4 and 5%, respectively) enabled the deconvolution of the DNA histogram and assignment of the proportion of nuclei into cell cycle compartments G(0) -G(1), S (DNA synthesis) and G(2) -M (Gap2-Mitosis). This technique can be also used for individual fish tissues such as brain, liver, fin and muscle. For the first time, the combined proportion of replicating nuclei (into S and G(2) -M phases) of whole fish larvae and absolute growth rate in length (mm day(-1)) has been correlated in commercial aquaculture conditions. Fast growing L. calcarifer larvae had an overall hyperplasia advantage as indicated by a greater proportion of cells in the S+G(2) -M phase compared with slow growing larvae, which might explain the increasing differences in size during culture. In a fasting trial, larvae ceased growth while maintaining the constant initial rates of cell division throughout a 6 day period. For a highly fed fast growing control group, cell division rates significantly increased after day 4. Flow-cytometric cell cycle analysis of whole fish larvae may provide fish biologists and aquaculturists with a better understanding of how cell division rates influence early growth in natural and artificial environments.     

Protein transduction in human cells is enhanced by cell-penetrating peptides fused with an endosomolytic HA2 sequence

Endocytosis has been proposed as one of the primary mechanisms for cellular entry of cell-penetrating peptides (CPPs) and their cargoes. However, a major limitation of endocytic pathway is entrapment of the CPP-cargo in intracellular vesicles from which the cargo must escape into the cytoplasm to exert its biological activity. Here we demonstrate that a CPP tagged with an endosomolytic fusion peptide derived from the influenza virus hemagglutinin-2 (HA2) remarkably enhances the cytosolic delivery of proteins in human A549 cells. To determine the endosome-disruptive effects, recombinant DNA plasmids containing coding sequences of HA2, CPPs and red fluorescent proteins (RFPs) were constructed. The fusion proteins were purified from plasmid-transformed Escherichia coli, and their effects on protein transduction were examined using live cell imaging and flow cytometry. Our data indicate that endocytosis is the major route for cellular internalization of CPP-HA2-tagged RFP. Mechanistic studies revealed that the fusogenic HA2 peptide dramatically facilitates CPP-mediated protein entry through the release of endocytosed RFPs from endosomes into the cytoplasm. Furthermore, incorporating the HA2 fusion peptide of the CPP-HA2 fusion protein improved cytosolic uptake without causing cytotoxicity. These findings strongly suggest that the CPP-HA2 tag could be an efficient and safe carrier that overcomes endosomal entrapment of delivered therapeutic drugs.     

Arginine vasopressin 1a receptor gene and maternal behavior: evidence of association and moderation

This study examined associations among maternal sensitivity, mothers' early adversity and the Arginine Vasopressin 1a Receptor (AVPR1A) gene. Early adversity in mothers' background has been found to be associated with lower maternal sensitivity. Animal literature suggests that variation in the AVPR1A gene is associated with parenting quality. The goal of the study was to examine the role of the AVPR1A gene in maternal sensitivity, especially under conditions of high early adversity. Participants included 151 Caucasian women from a community sample. The women were videotaped in their home while interacting separately with two of their children (target child = 18 months, older sibling <6 years). Evidence was found for an association between the AVPR1A gene and maternal sensitivity. Mothers with two copies of the long RS3 alleles were less sensitive than mothers with one or zero copies of the long alleles. This association was strongest under conditions of high maternal early adversity.     

Temporal priority and intra-guild predation in temporary waters: an experimental study using Namibian desert dragonflies

Abstract. 1. Intra-guild predation between early larval instars of two co-existing dragonflies, Sympetrum fonscolombii and Trithemis kirbyi , was investigated with respect to temporal advantage and growth. Three situations were simulated experimentally: (1) Sympetrum fonscolombii began development 11 days before T. kirbyi . (2) Trithemis kirbyi began development 11 days before S. fonscolombii . (3) Both species began on the same day.
2. With a temporal advantage of 11 days to the second species, the resulting larval density of the respective first species was significantly higher than that of the second species.
3. Without a temporal advantage, the survival of S. fonscolombii was higher than that of T. kirbyi , and S. fonscolombii had a larger size due to faster growth than T. kirbyi . Hence, it is assumed that survival depended on early oviposition as well as on larval growth.
4. To test the relevance of the laboratory results, observations at artificial ponds in the Namibian semi-desert were conducted. Trithemis kirbyi was the first species colonising these ponds while S. fonscolombii arrived 15 days later. In field samples, many more Trithemis larvae than Sympetrum larvae were found, a pattern similar to the laboratory experiments in which T. kirbyi enjoyed a temporal advantage.     

Flood-mediated dispersal versus hatching: early recolonisation strategies of copepods in floodplain ponds

1. We explored the quantitative contribution of two alternative recolonisation strategies by a combination of experimental and field data from the Little Tallahatchie floodplain (Mississippi, U.S.A.). In this area, several floods of short duration occur in winter and spring. During floods, lentic and lotic waterbodies of different size are connected for a limited period. 2. Shortly before the onset of the flood season we collected sediment samples from six temporary ponds. The emergence of seven cyclopoid species was recorded from the experimentally flooded sediments and was quantified over a 4‐week period. Water samples were taken in the same ponds after flooding, from which eight further species were recorded and quantified. 3. The contribution of flood‐dispersed individuals was variable among flooded ponds (between 38 and 94%), but numbers of flood‐dispersed colonists did not differ significantly from those of hatching individuals in flooded ponds. Our results demonstrate the equal importance of both hydrological connections and hatching from dormant stages as pathways for recolonisation by copepods if ponds are hydrologically connected. 4. The significance of recolonisation by hatching individuals in disconnected waterbodies was apparent in an isolated temporary pond that was exclusively recolonised by cyclopoid copepods emerging from the sediment.     

Maternally expressed PGK-Cre transgene as a tool for early and uniform activation of the Cre site-specific recombinase

A transgenic mouse strain with early and uniform expres sion of the Cre site-specific recombinase is described. In this strain, PGK-Crem, Cre is driven by the early acting PGK-1 promoter, but, probably due to cis effects at the integration site, the recombinase is under dominant maternal control. When Cre is transmitted by PGK-Crem females mated to males that carry a reporter transgene flanked by loxP sites, even offspring that do not inherit PGK-Cre delete the target gene. It follows that in the PGK-Crem female Cre activity commences in the diploid phase of oogenesis. In PGK-Crem crosses complete recombination was observed in all organs, including testis and ovary. We prepared a mouse stock that is homozygous for PGK-Crem and at the albino (c) locus. This strain will be useful for the early and uniform induction of ectopic and dominant negative mutations, for the in vivo removal of selective elements from targeted mutations and in connection with the manipulation of targeted loci in 'knock in' and related technologies