Production of biopesticide azadirachtin using plant cell and hairy root cultures

The extensive use of nondegradable chemical pesticides for pest management has developed serious environmental hazards. This has necessitated the urgent need to switch over to an alternative mode of biopesticide development for mass agriculture and field crop protection. Azadirachta indica A. Juss (commonly known as neem) houses a plethora of bioactive secondary metabolites with azadirachtin being the most active constituent explored in the sector of ecofriendly and biodegradable biopesticides characterized by low toxicity toward nontarget organisms. It has been reported that the highest content of azadirachtin and related limonoids is present in the seeds, available once in a year. Moreover, the inconsistent content and purity of the metabolites in whole plant makes it imperative to tap the potential of in vitro plant tissue culture applications, which would allow for several controlled manipulations for better yield and productivities. This review gives a summarized literature of the applied research and achievements in plant cell/hairy cultures of A. indica A. Juss mainly in context with the biopesticide azadirachtin and applications thereof.     

在体单向肠灌流模型研究白芷中三种成分的吸收特性

目的:研究白芷中有效成分欧前胡素、异欧前胡素和花椒毒酚在大鼠小肠各段的吸收特征。方法:采用大鼠在体单向肠灌流模型结合HPLC法同时测定肠灌流液中欧前胡素、异欧前胡素和花椒毒酚含量的变化,考察高、中、低三组剂量下三种成分在十二指肠、空肠和回肠的吸收特性。结果:三种成分随着剂量的增大,吸收速率常数(Ka)和有效渗透系数(Peff)逐渐增大。三种成分相同剂量下各肠段间吸收均无显著性差异,其吸收大小顺序为:欧前胡素异欧前胡素花椒毒酚。结论:白芷三种主要成分在各肠段均吸收良好,吸收大小呈剂量相关性。     

Systems pharmacology and enhanced pharmacodynamic models for understanding antibody-based drug action and toxicity

Pharmacokinetic (PK) and pharmacodynamic (PD) models seek to describe the temporal pattern of drug exposures and their associated pharmacological effects produced at micro- and macro-scales of organization. Antibody-based drugs have been developed for a large variety of diseases, with effects exhibited through a comprehensive range of mechanisms of action. Mechanism-based PK/PD and systems pharmacology models can play a major role in elucidating and integrating complex antibody pharmacological properties, such as nonlinear disposition and dynamical intracellular signaling pathways triggered by ligation to their cognate targets. Such complexities can be addressed through the use of robust computational modeling techniques that have proven powerful tools for pragmatic characterization of experimental data and for theoretical exploration of antibody efficacy and adverse effects. The primary objectives of such multi-scale mathematical models are to generate and test competing hypotheses and to predict clinical outcomes. In this review, relevant systems pharmacology and enhanced PD (ePD) models that are used as predictive tools for antibody-based drug action are reported. Their common conceptual features are highlighted, along with approaches used for modeling preclinical and clinically available data. Key examples illustrate how systems pharmacology and ePD models codify the interplay among complex biology, drug concentrations, and pharmacological effects. New hybrid modeling concepts that bridge cutting-edge systems pharmacology models with established PK/ePD models will be needed to anticipate antibody effects on disease in subpopulations and individual patients.     

Generation of diverse neural cell types through direct conversion

A characteristic of neurological disorders is the loss of critical populations of cells that the body is unable to replace,thus there has been much interest in identifying methods of generating clinically relevant numbers of cells to replace those that have been damaged or lost.The process of neural direct conversion,in which cells of one lineage are converted into cells of a neural lineage without first inducing pluripotency,shows great potential,with evidence of the generation of a range of functional neural cell types both in vitro and in vivo,through viral and non-viral delivery of exogenous factors,as well as chemical induction methods.Induced neural cells have been proposed as an attractive alternative to neural cells derived from embryonic or induced pluripotent stem cells,with prospective roles in the investigation of neurological disorders,including neurodegenerative disease modelling,drug screening,and cellular replacement for regenerative medicine applications,however further investigations into improving the efficacy and safety of these methods need to be performed before neural direct conversion becomes a clinically viable option.In this review,we describe the generation of diverse neural cell types via direct conversion of somatic cells,with comparison against stem cell-based approaches,as well as discussion of their potential research and clinical applications.     

Plant species with extremely small populations (PSESP) in China: A seed and spore biology perspective

Approximately one fifth of the world's plants are at risk of extinction. Of these, a significant number exist as populations of few individuals, with limited distribution ranges and under enormous pressure due to habitat destruction. In China, these most-at-risk species are described as ‘plant species with extremely small populations’ (PSESP). Implementing conservation action for such listed species is urgent. Storing seeds is one of the main means of ex situ conservation for flowering plants. Spore storage could provide a simple and economical method for fern ex situ conservation. Seed and spore germination in nature is a critical step in species regeneration and thus in situ conservation. But what is known about the seed and spore biology (storage and germination) of at-risk species? We have used China's PSESP (the first group listing) as a case study to understand the gaps in knowledge on propagule biology of threatened plant species. We found that whilst germination information is available for 28 species (23% of PSESP), storage characteristics are only known for 8% of PSESP (10 species). Moreover, we estimate that 60% of the listed species may require cryopreservation for long-term storage. We conclude that comparative biology studies are urgently needed on the world's most threatened taxa so that conservation action can progress beyond species listing.     

Reproductive isolation between sympatric sister species,Mussaenda kwangtungensis and M. pubescens var. alba

Reproductive isolation de fi nes the biological species concept and plays a key role in the formation and maintenance of species. The relative contributions of different isolating stages has been suggested to be closely associated with phylogenetic relatedness. Few studies have focused on the relative contributions of pre- versus postzygotic mechanisms, and even fewer have been conducted under strict phylogenetic frameworks. Pre- and post-zygotic reproductive isolation stages have been investigated in the sister species Mussaenda kwangtungensis and M. pubescens var. alba. The two species have partly overlapping distribution ranges and fl owering times, while the principal pollinators differed strikingly for them, demonstrating strong pre-zygotic isolations. Natural hybrids were detected by simple sequence repeat markers and their maternal parents were identi fi ed based on chloroplast gene sequences. Five out of 81 individuals were suggested to be hybrids that fall into the categories F2, BC1, and BC2 by theNew Hybrids analysis. Interspeci fi c crossings resulted in signi fi cantly reduced fruit set and seed germination rates.Phylogenetic analysis revealed short Kimura-2-parameter distance between M. kwangtungensis and M. pubescens var.alba. These fi ndings strongly supported the hypothesis that for species with a closer phylogenetic relationship, prezygotic isolation plays an important part in limiting gene exchange in sympatric areas.     

Aegilops conservation and collection evaluation in the Czech Republic

Aegilops distribution was revised on localities in the territory of the Czech Republic and the former Czechoslovakia, based on herbarium data, literature and floristic databases. The only surviving species Ae. cylindrica forms the northern limit of distribution in the Danube basin, other sites are secondary. In situ conservation was proposed for two Czech sites where Aegilops have been naturalized. An ex situ gene bank collection has been maintained in the Gene Bank Prague since 1985. The collection includes 21 species and 1099 accessions. The whole collection was evaluated for breeding-valuable characters: resistance to leaf diseases, cereal aphids, viral diseases, and qualitative parameters. Recently, the collection was retested for the present races of leaf and stem rust including new introductions from Kazakhstan. Six different leaf rust races and five different stem rust races collected from the Czech Republic were applied in the older tests, and three races from each rust species in the recent tests. The highest number of accessions resistant both to wheat leaf rust and wheat stem rust and powdery mildew was found in Ae. speltoides (90%). Different reactions of individual accessions of the tested species to the three different pathotypes of each rust species indicates that several diverse genes for specific resistance are present in the accessions of the tested species.     

Taxonomic diversity within the Japanese green lacewing, Chrysoperla carnea (Neuroptera: Chrysopidae), identified by courtship song analyses and crossing tests

The green lacewing, Chrysoperla carnea (Stephens), is an important natural enemy of various crop pests, especially aphids. In the Japanese fauna, there are two types of larval forms, A and B, characterized by different head capsule markings. The Type A form is distributed throughout Japan, but the Type B form has a more limited distribution. Adults use abdominal vibration as a communication signal (courtship song) during mating. We analyzed oscillograms of these songs among several Japanese populations of C. carnea. The courtship songs of types A and B are distinctly different from one another. We then performed crossing tests between the two types. Copulation between same-type pairings was much more likely than between different-type pairings. We also analyzed courtship songs of European C. carnea sensu stricti, introduced to Japan as a natural enemy of crop pests. The song of these introduced green lacewings appeared to differ from either type of Japanese C. carnea. The two types of C. carnea are likely to be different species, and also distinct from C. carnea sensu stricti of Europe.     

Ex situ transportation of coral larvae for research,conservation, and aquaculture

Due to the lack of appropriate methods to transport high amounts of larvae ex situ over large distances, the availability of coral larvae was so far mainly limited to their place of origin. For a research project at Rotterdam Zoo, The Netherlands, we transported several thousand larvae of three broadcast spawners (Acropora tenuis, A. digitifera, Diploria strigosa) from the Indo Pacific and the Caribbean to Europe. Beside logistics and packing techniques, post-transport survival rates were mainly influenced by larvae density and transport duration. Our results indicate optimum survival rates of >90% at densities of 4 larvae ml⁻¹ when not exceeding a transportation time of 4 days. The ex situ transport of coral larvae over large distances might offer new possibilities for research, conservation, and aquaculture.     

肌肽对离体培养的鸡肝细胞的影响

研究了肌肽对离体培养的鸡肝细胞的影响。实验采用胶原酶原位二步灌流法获得鸡肝细胞,再用不同浓度的肌肽(①2 μmol L、②2 0 μmol L、③2 0 0 μmol L ,④2 0mmol L)分别进行处理。结果表明,1 )在各个时期,含2 0 μmol L肌肽的实验组肝细胞生长均好于对照组(0 μmol L肌肽) ,而且在48和72h时二者差异显著(P <0 0 5 ) ;2 ) 2 0mmol L肌肽能显著提高肝细胞的分泌白蛋白的功能(P <0 0 5 ) ;3 )添加肌肽的各实验组上清液中丙二醛(MDA)含量在2 4和48h时均低于对照组;4)添加2 0mmol L肌肽的实验组与对照组相比,能维持无血清培养肝细胞的形态达8d。表明了肌肽对鸡肝细胞增殖和分泌白蛋白有促进作用,同时可以保护肝细胞对抗过氧化,保护细胞形态。