Production of somatic embryos and shoots from sugarbeet callus: Effects of abscisic acid, other growth regulators, nitrogen source, sucrose concentration and genotype

Summary Two sugarbeet (Beta vulgaris L.) genotypes, REL-1 and REL-2, were used to measure the level of somatic embryo and shoot production from hormone-autonomous callus plated under varied nutrient medium combinations of abscisic acid with the growth regulators 6-benzyladenine, 1-naphthaleneacetic acid, or 2,4-dichlorophenoxyacetic acid, with eight sole nitrogen sources, or with different sucrose concentrations. Clone REL-2 produced embryos up to 35-fold more frequently than clone REL-1. Inclusion of abscisic acid at some concentrations consistently improved embryo production in all experiments and was observed to stimulate shoot production. At some concentrations, 1-naphthaleneacetic acid as well as urea and glutamine stimulated greater embryo production over the control, but only for REL-1, for which there was greater room for improvement. Three and five percent sucrose were superior to 1, 7, and 9%. Higher initial 6-benzyladenine concentration [in the range 0, 0.1−1.0 mg/L (0.44−4.44 μM)] was associated with lower embryo production but greater shoot regeneration for both clones. REL-2 was significantly better than REL-1 in shoot regeneration. The range of embryo production was more than 35-fold between genotypes, whereas the range of physiological effects was no greater than 10-fold. REL-2 has been released to sugarbeet researchers because of its superior embryogenic and shoot regeneration abilities for application in biotechnology.     

A sequential expression system for high-throughput functional genomic analysis

A method employing sequential rounds of cell-free protein synthesis (CFPS) was developed to identify gene products influencing the complex metabolic systems that result in protein accumulation and folding in vitro. The first round of CFPS creates an array of cell extracts individually enriched with a single gene product expressed in-parallel from linear DNA expression templates (ETs). The cell extract is engineered to enhance template stability and to provide reaction conditions conducive for general protein activation. Following first-round expression, linear templates are selectively degraded and a plasmid template for a reporter enzyme is added to initiate a subsequent round of protein expression. Reporter concentration and activity identify first-round gene products that affect amino acid and nucleic acid stability, energy supply, protein expression, stability, and activation. This sequential CFPS system provides a unique format for the functional genomic identification of broadly diverse metabolic activities.     

Structural understanding of stabilization patterns in engineered bispecific Ig‐like antibody molecules

Bispecific immunoglobulin‐like antibodies capable of engaging multiple antigens represent a promising new class of therapeutic agents. Engineering of these molecules requires optimization of the molecular properties of one of the domain components. Here, we present a detailed crystallographic and computational characterization of the stabilization patterns in the lymphotoxin‐beta receptor (LTβR) binding Fv domain of an anti‐LTβR/anti‐TNF‐related apoptosis inducing ligand receptor‐2 (TRAIL‐R2) bispecific immunoglobulin‐like antibody. We further describe a new hierarchical structure‐guided approach toward engineering of antibody‐like molecules to enhance their thermal and chemical stability. Proteins 2009. © 2009 Wiley‐Liss, Inc.     

Quantifying the evidence for ecological synergies

There is increasing concern that multiple drivers of ecological change will interact synergistically to accelerate biodiversity loss. However, the prevalence and magnitude of these interactions remain one of the largest uncertainties in projections of future ecological change. We address this uncertainty by performing a meta-analysis of 112 published factorial experiments that evaluated the impacts of multiple stressors on animal mortality in freshwater, marine and terrestrial communities. We found that, on average, mortalities from the combined action of two stressors were not synergistic and this result was consistent across studies investigating different stressors, study organisms and life-history stages. Furthermore, only one-third of relevant experiments displayed truly synergistic effects, which does not support the prevailing ecological paradigm that synergies are rampant. However, in more than three-quarters of relevant experiments, the outcome of multiple stressor interactions was non-additive (i.e. synergies or antagonisms), suggesting that ecological surprises may be more common than simple additive effects.     

Biochemical and strain properties of CJD prions: complexity versus simplicity

Prions, the agents responsible for transmissible spongiform encephalopathies, are infectious proteins consisting primarily of scrapie prion protein (PrP(Sc)), a misfolded, β-sheet enriched and aggregated form of the host-encoded cellular prion protein (PrP(C)). Their propagation is based on an autocatalytic PrP conversion process. Despite the lack of a nucleic acid genome, different prion strains have been isolated from animal diseases. Increasing evidence supports the view that strain-specific properties may be enciphered within conformational variations of PrP(Sc). In humans, sporadic Creutzfeldt-Jakob disease (sCJD) is the most frequent form of prion diseases and has demonstrated a wide phenotypic and molecular spectrum. In contrast, variant Creutzfeldt-Jakob disease (vCJD), which results from oral exposure to the agent of bovine spongiform encephalopathy, is a highly stereotyped disease, that, until now, has only occurred in patients who are methionine homozygous at codon 129 of the PrP gene. Recent research has provided consistent evidence of strain diversity in sCJD and also, unexpectedly enough, in vCJD. Here, we discuss the puzzling biochemical/pathological diversity of human prion disorders and the relationship of that diversity to the biological properties of the agent as demonstrated by strain typing in experimental models.     

Flexible Solar Thermal Fuel Devices: Composites of Fabric and a Photoliquefiable Azobenzene Derivative

Controllable storage and release of solar energy has always been a highlighted scientific issue for its benefit of mankind. Solar thermal fuels (STFs) supply a closed cycle and renewable energy‐storage strategy by transforming solar energy into chemical energy stored in the conformation of molecular isomers, such as cis/trans‐azobenzene, and releasing it as heat under various stimuli. Although the potential high energy density of the STFs which are based on the hybrids of azobenzene derivatives and carbon nanomaterials has been reported the solvent‐assistant charging hinders their practicability. In this study, a solid‐state STF device is designed and fabricated by compositing one photoliquefiable azobenzene (PLAZ) derivative with a flexible fabric template. The photoinduced phase transition of the PLAZ derivative enables the charging of the flexible STFs to be totally solvent‐free. Interestingly, the energy‐storage capacity (energy density ≈201 J g^?1) of flexible PLAZ STFs has been improved by the soft fabric template. The exothermic situation is monitored with one infrared camera, which shows 4 °C temperature difference between charged and discharged samples under blue light stimulus. The flexible STFs are may be used in practice as heating equipment.     

蝉花虫草活性成分的抗惊厥作用

本研究采用活性追踪的方法,逐步从人工培养蝉花虫草分离获得A（50%乙醇回流提取）、B（膜分离）、C（大孔树脂洗脱）和D（Sephadex LH20柱纯化）等4种样品,单一化合物D纯度为98.62%,鉴定为N6‐(2‐羟乙基)腺苷[N6‐(2‐hydroxyethyl)‐adenosine,HEA]。研究各样品对戊四唑（pentylenetetrazol,PTZ）诱导的小鼠惊厥模型的影响,以及选择性腺苷A1受体（AA1R）拮抗剂DPCPX或选择性腺苷A2A受体（AA2AR）拮抗剂ZM241385对HEA作用的影响,并采用苏木精-伊红（hematoxylin-eosin,HE）染色、免疫组化（immunohistochemical,IHC）染色和蛋白质免疫印迹（Western blot,WB）等技术进一步探究HEA抗惊厥作用的机制。结果表明,各样品（i.p.）均有抗惊厥活性,HEA（40–60mg/kg,i.p.）能显著延长惊厥小鼠的存活时间和降低死亡率,DPCPX（2mg/kg,i.p.）能够拮抗HEA的抗惊厥作用,而ZM241385无此作用;HE、IHC和WB的结果进一步揭示DPCPX显著降低HEA的作用。综上所述,蝉花虫草的HEA具有抗惊厥作用,并且可能通过激活腺苷A1受体而起作用。     

Developing a vision for improved monitoring and reporting of riparian restoration projects

Representatives from agencies involved in natural resource management in the Murray‐Darling Basin gathered for a workshop in November 2010 to develop a vision for improved monitoring and reporting of riparian restoration projects. The resounding message from this workshop was that the effectiveness of riparian restoration depends on having sound, documented and agreed evidence on the ecological responses to restoration efforts. Improving our capacity to manage and restore riparian ecosystems is constrained by (i) a lack of ecological evidence on the effects of restoration efforts, and (ii) short‐termism in commitment to restoration efforts, in funding of monitoring and in expected time spans for ecosystem recovery. Restoration at the effective spatial scope will invariably require a long‐term commitment by researchers, funding agencies, management agencies and landholders. To address the knowledge gaps that constrain riparian restoration in the Basin, participants endorsed four major fields for future research: the importance of landscape context to restoration outcomes; spatio‐temporal scaling of restoration outcomes; functional effects of restoration efforts; and developing informative and effective indicators of restoration. To improve the monitoring and restoration of riparian zones throughout the Basin, participants advocated an integrated approach: a hierarchical adaptive management framework that incorporates long‐term ecological research.