Strategies for the identification and purification of ligands for orphan biomolecules

The isolation of related genes with evolutionary conserved motifs by the application ofpolymerase chain reaction-based molecular biology techniques, or from database searchingstrategies, has facilitated the identification of new members of protein families. Many of theseprotein molecules will be involved in protein–protein interactions (e.g. growth factors,receptors, adhesion molecules), since such interactions are intrinsic to virtually every cellularprocess. However, the precise biological function and specific binding partners of these novelproteins are frequently unknown, hence they are known as orphan molecules.Complementary technologies are required for the identification of the specific ligands orreceptors for these and other orphan proteins (e.g., antibodies raised against crude biologicalextracts or whole cells). We describe herein several alternative strategies for the identification,purification and characterisation of orphan peptide and protein molecules, specifically thesynergistic use of micropreparative HPLC and biosensor techniques.     

绵羊粒细胞集落刺激因子原核表达与提纯及用于颗粒细胞培养的效果

文中旨在研究粒细胞集落刺激因子(Granule cell stimulating factor,GCSF)对绵羊颗粒细胞体外培养过程中细胞增殖和凋亡的影响,明确GCSF对绵羊颗粒细胞生存的调节作用,为今后该蛋白用于羊繁育方面的研究奠定基础。原核克隆表达纯化羊GCSF蛋白,纯化的蛋白用M-NSF60细胞检测生物学活性,将纯化的GCSF添加到颗粒细胞培养基中作为试验组,以培养基中未添加GCSF的细胞为对照,利用Alarmarblue检测细胞增殖情况,流式细胞仪检测细胞周期和凋亡的变化。结果表明,羊GCSF可以原核表达并纯化,并且具有生物学活性。在24 h和48 h时,在0.06–600 ng/mL的范围内随着加入GCSF的终浓度增加,细胞活力升高。试验组颗粒细胞体外培养24 h后,与阴性对照相比,细胞周期的分布显著改变,S期细胞比例显著减少(P<0.05),G2/M期细胞比例显著增多(P<0.05)。试验组凋亡率和对照组相比,48 h检测时凋亡率显著降低(P<0.05)。综上表明,GCSF在体外培养的绵羊颗粒细胞中,可调控绵羊颗粒细胞周期,促进细胞增殖,抑制细胞凋亡。     

In vitro biological evaluation of platinum(II) complexes with 1-(methoxy substituted benzyl) azetidine-3,3-dicarboxylato ligands

A number of platinum(II) complexes with ammine or 1R,2R-diaminocyclohexane as carrier ligands and 1-(methoxy-substituted benzyl) azetidine-3,3-dicarboxylate as leaving groups were synthesized and spectrally characterized. Biological evaluation in vitro showed that some of compounds showed positive antitumor activity. In particular, complex 3a, (1R,2R-diaminocyclohexane)[1-(3-methoxylbenzyl) azetidine-3,3-dicarboxylato)-O,O'] platinum(II), possessed a potent antitumor effect comparable to cisplatin and/or oxaliplatin, and very low toxicity in vivo. Preliminary antitumor mechanism of 3a has been investigated by cell apoptosis assays compared with cisplatin and oxaliplatin.     

Immunological features and efficacy of a multi-epitope vaccine CTB-UE against H. pylori in BALB/c mice model

Epitope vaccine is a promising option for prophylactic and therapeutic vaccination against Helicobacter pylori infection. Urease is an essential virulence factor and colonization factor for H. pylori. In this study, we constructed a multi-epitope vaccine named CTB-UE with mucosal adjuvant cholera toxin B subunit (CTB) and tandem copies of Th and B cell epitopes from H. pylori urease A and B subunits. The immunogenicity, specificity, ability to induce neutralizing antibodies against H. pylori urease, and prophylactic and therapeutic efficacy of the CTB-UE vaccine were evaluated in BALB/c mice model after purification. The experimental results indicated that CTB-UE could induce comparatively high levels of specific antibodies against native H. pylori urease, UreA, UreB, or the selected B cell epitopes UreA_183–203 and UreB_327–334 involved with the active site of urease and showed an effectively inhibitory effect on the enzymatic activity of urease. Besides, oral prophylactic or therapeutic immunization with CTB-UE significantly decreased H. pylori colonization compared with oral immunization with rUreB or PBS, and the protection was correlated with antigen-specific CD4⁺ T cells and IgG, IgA, and mucosal sIgA antibody responses. This CTB-UE vaccine may be a promising vaccine candidate for the control of H. pylori infection.     

露天煤矿排土场边坡植被组成特征及其群落稳定性评价

研究植被恢复过程中植物群落组成、结构及稳定性的变化,可进一步了解到植物群落的演替过程及规律。采用空间代时间的方法对内蒙古锡林郭勒盟北电胜利露天煤矿4个排土场边坡人工恢复植被进行群落调查,在此基础上采用多样性指数与优化后的M.Godron稳定性指数对植物物种组成、物种多样性及其群落稳定性进行分析,探究不同恢复年限排土场边坡植被在恢复过程中群落特征及稳定性变化特征。结果表明：（1）排土场边坡植物物种组成共有16科44属56种,植物群落组成较简单,其中禾本科种类最多,生长型以草本植物为主。（2）随着恢复年限的增加,植物生活型由一、二年生转变为多年生,坡面优势种由人工种植植物转变为本土植物。（3）整体看,随着恢复年限增加,物种多样性呈下降趋势,群落稳定性整体呈上升趋势。（4）由于不同坡向土壤微环境差异,北坡恢复效果及稳定性好于南坡。本研究在一定程度上可为露天煤矿排土场边坡人工修复过程中植物物种及合理配置模式的选择提供科学依据。     

Strategies for the identification and purification of ligands for orphan biomolecules

Summary The isolation of related genes with evolutionary conserved motifs by the application of polymerase chain reaction-based molecular biology techniques, or from database searching strategies, has facilitated the identification of new members of protein families. Many of these protein molecules will be involved in protein-protein interactions (e.g. growth factors, receptors, adhesion molecules), since such interactions are intrinsic to virtually every cellular process. However, the precise biological function and specific binding partners of these novel proteins are frequently unknown, hence they are known as ‘orphan’ molecules. Complementary technologies are required for the identification of the specific ligands or receptors for these and other orphan proteins (e.g., antibodies raised against crude biological extracts or whole cells). We describe herein several alternative strategies for the identification, purification and characterisation of orphan peptide and protein molecules, specifically the synergistic use of micropreparative HPLC and biosensor techniques. These authors made equivalent contributions.     

Does more mean less? The value of information for conservation planning under sea level rise

Many studies have explored the benefits of adopting more sophisticated modelling techniques or spatial data in terms of our ability to accurately predict ecosystem responses to global change. However, we currently know little about whether the improved predictions will actually lead to better conservation outcomes once the costs of gaining improved models or data are accounted for. This severely limits our ability to make strategic decisions for adaptation to global pressures, particularly in landscapes subject to dynamic change such as the coastal zone. In such landscapes, the global phenomenon of sea level rise is a critical consideration for preserving biodiversity. Here, we address this issue in the context of making decisions about where to locate a reserve system to preserve coastal biodiversity with a limited budget. Specifically, we determined the cost‐effectiveness of investing in high‐resolution elevation data and process‐based models for predicting wetland shifts in a coastal region of South East Queensland, Australia. We evaluated the resulting priority areas for reserve selection to quantify the cost‐effectiveness of investment in better quantifying biological and physical processes. We show that, in this case, it is considerably more cost effective to use a process‐based model and high‐resolution elevation data, even if this requires a substantial proportion of the project budget to be expended (up to 99% in one instance). The less accurate model and data set failed to identify areas of high conservation value, reducing the cost‐effectiveness of the resultant conservation plan. This suggests that when developing conservation plans in areas where sea level rise threatens biodiversity, investing in high‐resolution elevation data and process‐based models to predict shifts in coastal ecosystems may be highly cost effective. A future research priority is to determine how this cost‐effectiveness varies among different regions across the globe.     

Antitumor activities and interaction with DNA of oxaliplatin-type platinum complexes with linear or branched alkoxyacetates as leaving groups

Five oxaliplatin-typed platinum complexes containing trans-1R, 2R-diaminocyclohexane chelating platinum cores, characteristic of linear or branched alkoxycarboxylates as leaving groups, were biologically evaluated. These compounds showed higher antitumor activity, lower toxicity in vivo than cisplatin or oxaliplatin. And the results revealed that the antitumor activity and interaction with DNA of these compounds were highly related to the nature of leaving groups. Among these complexes, 5a, cis-(trans-1R, 2R-diaminocyclohexane) bis (2-tert-butoxyacetate) platinum(II), showed the highest antitumor activity and the lowest toxicity.