Development of a Blocking ELISA for Detection of Serum Neutralizing Antibodies against Newly Emerged Duck Tembusu Virus

Background

Since April 2010, domesticated ducks in China have been suffering from an emerging infectious disease characterized by retarded growth, high fever, loss of appetite, decline in egg production, and death. The causative agent was identified as a duck Tembusu virus (DTMUV), a member of the Ntaya virus (NTAV) group within the genus Flavivirus, family Flaviviridae. DTMUV is highly contagious and spreads rapidly in many species of ducks. More than 10 million shelducks have been infected and approximately 1 million died in 2010. The disease remains a constant threat to the duck industry; however, it is not known whether DTMUV can infect humans or other mammalians, despite the fact that the virus has spread widely in southeast China, one of the most densely populated areas in the world. The lack of reliable methods to detect the serum antibodies against DTMUV has limited our ability to conduct epidemiological investigations in various natural hosts and to evaluate the efficiency of vaccines to DTMUV.

Methodology/Principal Findings

A neutralizing monoclonal antibody (mAb) 1F5 binding specifically to the E protein was developed. Based on the mAb, a blocking enzyme-linked immunosorbent assay (ELISA) was developed for the detection of neutralizing antibodies against DTMUV. The average value of percent inhibition (PI) of 350 duck serum samples obtained from DTMUV-free farms was 1.0% ±5.8% (mean ± SD). The selected cut-off PI values for negative and positive sera were 12.6% (mean +2SD) and 18.4% (mean +3SD), respectively. When compared with a serum neutralizing antibody test (SNT) using chicken embryonated eggs, the rate of coincidence was 70.6% between the blocking ELISA and SNT, based on the titration of 20 duck DTMUV-positive serum samples.

Conclusions/Significance

The blocking ELISA based on a neutralizing mAb allowed rapid, sensitive, and specific detection of neutralization-related antibodies against DTMUV.     

Electrochemical detection of dopamine using porphyrin-functionalized graphene

A new type of porphyrin-functionalized graphene was synthesized and used for highly selective and sensitive detection of dopamine (DA). The aromatic π-π stacking and electrostatic attraction between positively-charged dopamine and negatively-charged porphyrin-modified graphene can accelerate the electron transfer whereas weakening ascorbic acid (AA) and uric acid (UA) oxidation on the porphyrin-functionalized graphene-modified electrode. Differential pulse voltammetry was used for electrochemical detection, the separation of the oxidation peak potentials for AA-DA, DA-UA and UA-AA is about 188 mV, 144 mV and 332 mV, which allows selectively determining DA. The detection limit of DA can be as low as 0.01 μM. More importantly, the sensor we presented can detect DA in the presence of large excess of ascorbic acid and uric acid. With good sensitivity and selectivity, the present method was applied to the determination of DA in real hydrochloride injection sample, human urine and serum samples, respectively, and the results was satisfactory.     

Comparative proteomic study reveals dynamic proteome changes between superhybrid rice LYP9 and its parents at different developmental stages

Heterosis is a common phenomenon in which the hybrids exhibit superior agronomic performance than either inbred parental lines. Although hybrid rice is one of the most successful apotheoses in crops utilizing heterosis, the molecular mechanisms underlying rice heterosis remain elusive. To gain a better understanding of the molecular mechanisms of rice heterosis, comparative leaf proteomic analysis between a superhybrid rice LYP9 and its parental cultivars 9311 and PA64s at tillering, flowering and grain-filling stages were carried out. A total of 384 differentially expressed proteins (DP) were detected and 297 DP were identified, corresponding to 222 unique proteins. As DP were divided into those between the parents (DP_PP) and between the hybrid and its parents (DP_HP), the comparative results demonstrate that proteins in the categories of photosynthesis, glycolysis, and disease/defense were mainly enriched in DP. Moreover, the number of identified DP_HP involved in photosynthesis, glycolysis, and disease/defense increased at flowering and grain-filling stages as compared to that at the tillering stage. Most of the up-regulated DP_HP involved in the three categories showed greater expression in LYP9 at flowering and grain-filling stages than at the tillering stage. In addition, CO₂ assimilation rate and apparent quantum yield of photosynthesis also showed a greater increase in LYP9 at flowering and grain-filling stages than at the tillering stage. These results suggest that the proteins involved in photosynthesis, glycolysis, and disease/defense as well as their dynamic regulation at different developmental stages may be responsible for heterosis in rice.     

A reversible DNA-silver nanoclusters-based molecular fluorescence switch and its use for logic gate operation

Molecule-like silver nanoclusters (AgNCs) with few to tens of atoms are highly sensitive to the sequence and structure of DNA stabilizers. In this paper, a novel pH-triggered reversible molecular fluorescence switch is developed by taking advantage of the DNA-dependent fluorescence pH response of AgNCs. The DNA-AgNCs fluorescence switch simultaneously addresses concerns of simple construction strategy, efficient design and organic-solvent-free operation. Moreover, the excellent photostability and biocompatibility of AgNCs provide great potential for application of the DNA-AgNCs fluorescence switch in the development of functional molecular devices. Specifically, we apply the DNA-AgNCs fluorescence switch combined with the DNA sequence-dependent pH response pattern of AgNCs for construction of molecular logic gates.     

敦煌阳关湿地芦苇克隆构件空间拓展策略及其对土壤环境因子的响应

空间拓展是克隆植物响应环境的生态适应和权衡维度,以内陆盐沼湿地典型代表敦煌阳关国家级自然保护区内的芦苇(Phragmites australis)为研究对象,基于水库距离远近和芦苇种群的密度与盖度,设置了近水区(500 m)、过渡区(1500 m)、荒漠区(2500 m)3个不同的采样梯度,研究了环境异质条件下芦苇克隆构件空间拓展策略及其对土壤环境因子的响应。结果表明:(1)芦苇克隆构件在异质环境条件下差异显著,初级根茎条数、根茎节间长、间隔子长、分枝角度呈现"同增同涨"的协同进化关系(P0.01),而与分株数呈现"此消彼长"的权衡关系(P0.01)。(2)且随着种群的密度降低,芦苇在空间拓展过程中呈现从"密集型"转"游击型"的生存策略。(3)冗余分析显示土壤水分、pH值、盐分是克隆构件空间拓展的主要驱动力,其中各层土壤水分、深层土壤pH值和盐分是芦苇空间拓展最重要的环境因子。明晰内陆河湿地芦苇克隆构件空间拓展策略及主要的环境驱动力,能为干旱脆弱敏感区湿地保护和生态恢复提供借鉴意义。     

“在哪里”和“有多少”? 中国雪豹调查与空缺

雪豹(Panthera uncia)分布广泛且调查难度较大, 全世界的雪豹研究面临的首要问题是雪豹基础数据的缺乏。本文通过检索1980至2018年已发表的含有中国境内雪豹分布和密度信息的中英文文章共35篇, 从中提取出雪豹的分布与密度信息, 其中含有密度估计的文献18篇。同时, 来自雪豹调查的15位一线成员通过填写表格和问卷的形式提供了28个地块上未发表的雪豹密度调查信息。基于此, 我们逐一分析了各省份已有的雪豹调查现状和存在的调查空缺, 发现雪豹分布调查的两大空白区域主要存在于与吉尔吉斯斯坦接壤的西天山地区和西藏南部的冈底斯-念青唐古拉山山脉和喜马拉雅山脉。相对我国雪豹栖息地总面积, 有过密度估算的面积仅占1.7%, 仍然处于刚刚起步的阶段, 并且已有的密度调查几乎都是在质量较好的雪豹栖息地内进行的。今后除了需要继续努力收集汇总已有的调查结果, 仍然需要在雪豹分布区(特别是空缺区域)内增加调查。     

不同生态条件下油松(Pinus tabulaeformis)菌根根际土壤微生物群落

利用BIOLOG代谢指纹方法分析了陕南商南和陕北安塞不同生态条件下油松菌根根际土壤微生物群落。结果表明,安塞油松和商南油松菌根根际微生物对糖类和氨基酸类碳源较易利用,商南油松菌根根际微生物总体上代谢碳源的种类和活性远大于安塞油松,而且对同类碳源的代谢商南油松的AWCD比安塞油松均高出2倍多。安塞油松菌根根际微生物以氨基酸类代谢群为优势类群,商南油松以糖类代谢群为优势类群。微生物群落多样性指数和微生物群落主成分分析（PCA）指标均表明商南油松和安塞油松菌根根际土壤微生物群落有明显不同,起分异作用的碳源主要为糖类,其次是羧酸类和氨基酸类。商南油松菌根根际土壤微生物群落AWCD极显著高于安塞油松（P<0.01）,细菌数量显著高于安塞油松（P<0.05）,Shannon指数和丰富度指数达极显著性差异（P<0.01）,商南油松和安塞油松菌根侵染率差异不显著,但菌根生物量差异达极显著水平（P<001）。相关性分析表明,菌根生物量与丰富度指数、AWCD呈极显著正相关,与Shannon指数呈显著正相关,但是与菌根侵染率相关性不显著。在商南温暖潮湿丘陵区油松菌根根际微生物活性、群落大小和多样性高于安塞油松,在安塞黄土高原干旱区微生物群落稳定性强于商南油松。     

重金属污染对红壤旱地小节肢类土壤动物群落结构的影响

土壤重金属污染对农田土壤节肢动物的影响是其环境风险评价和监测的重要方面。基于现有国家土壤环境质量二级标准(GB 15618—1995),设计2个浓度重金属Cd(0.3 mg/kg,0.6 mg/kg)、Pb(125 mg/kg,250 mg/kg)和As(30 mg/kg,60 mg/kg)污染的田间试验,分别研究Cd、Pb和As对红壤旱地小节肢类土壤动物群落组成和多样性的影响。结果表明,试验土壤小节肢类动物共11类群,其中符跳属、棘跳属、蜱螨目为本地区红壤旱地的主要类群,占全部捕获小节肢类土壤动物总数的78%。重金属Pb、As高浓度污染分别显著降低了土壤弹尾目的个体密度73.5%、55.2%,而Cd高浓度污染对其无显著影响,仅降低15.9%;As和Cd高浓度污染分别显著降低了土壤蜱螨目的个体密度74.3%、36.4%,而Pb高浓度污染对其无显著影响。Pb、As和Cd污染对小节肢类土壤动物群落多样性指数均无明显影响。因此,与丰度相比,多样性指数对重金属污染反应的灵敏度较差。主成分分析表明,前气门亚目、符跳属和棘跳属的得分值较高,为小节肢类土壤动物群落中的主导因子,可作为未来该地区重金属污染环境影响监测的重要指标生物和重金属污染阈值研究的重要生态受体。     

基于主成分分析的广西省干旱时空格局

广西省地处喀斯特地貌区,土壤保水效率低,且年降水时空分布不均,研究干旱时空分布尤为重要。基于广西省1981—2010年20个气象站实测和2011—2100年HadGEM2-ES模型模拟数据,利用标准化降水指数(SPI, Standardized Precipitation Index)和标准化降水蒸散指数(SPEI,Standardized Precipitation Evapotranspiration Index),分析广西省干旱的时空变化。对不同时间尺度的SPI和SPEI应用主成分分析(PCA,Principal Component Analysis)确定干旱的空间模式,结果揭示了3个空间分布明确的区域:桂东北地区(PC1),桂西北地区(PC2)和桂南地区(PC3)。各区域干旱的时空变化和频率分布差异显著。PC1和PC3的SPEI-12呈负增长趋势,PC2的SPEI-12呈正增长趋势,且PC1、PC2和PC3的SPI-12均大于SPEI-12。年尺度(SPEI-12)上PC1和PC3的干旱频率大于PC2,其干旱频率分别为34.24%和35.83%。SPI和SPEI在空间和时间尺度...     

Mitochondrial DNA associations with East Asian metabolic syndrome

Mitochondrial dysfunction has repeatedly been reported associated with type 2 diabetes mellitus (T2DM) and metabolic syndrome (MS), as have mitochondrial DNA (mtDNA) tRNA and duplication mutations and mtDNA haplogroup lineages. We identified 19 Taiwanese T2DM and MS pedigrees from Taiwan, with putative matrilineal transmission, one of which harbored the pathogenic mtDNA tRNA^Leu(UUR) nucleotide (nt) 3243A>G mutation on the N9a3 haplogroup background. We then recruited three independent Taiwanese cohorts, two from Taipei (N?=?498, mean age 52 and N?=?1002, mean age 44) and one from a non-urban environment (N?=?501, mean age 57). All three cohorts were assessed for an array of metabolic parameters, their mtDNA haplogroups determined, and the haplogroups correlated with T2DM/MS phenotypes. Logistic regression analysis revealed that mtDNA haplogroups D5, F4, and N9a conferred T2DM protection, while haplogroups F4 and N9a were risk factors for hypertension (HTN), and F4 was a risk factor for obesity (OB). Additionally, the 5263C>T (ND2 A165V) variant commonly associated with F4 was associated with hypertension (HTN). Cybrids were prepared with macro-haplogroup N (defined by variants m.ND3 10398A (114T) and m.ATP6 8701A (59T)) haplogroups B4 and F1 mtDNAs and from macro-haplogroup M (variants m.ND3 10398G (114A) and m.ATP6 8701G (59A)) haplogroup M9 mtDNAs. Additionally, haplogroup B4 and F1 cybrids were prepared with and without the mtDNA variant in ND1 3394T>C (Y30H) reported to be associated with T2DM. Assay of mitochondria complex I in these cybrids revealed that macro-haplogroup N cybrids had lower activity than M cybrids, that haplogroup F cybrids had lower activity than B4 cybrids, and that the ND1 3394T>C (Y30H) variant reduced complex I on both the B4 and F1 background but with very different cumulative effects. These data support the hypothesis that functional mtDNA variants may contribute to the risk of developing T2DM and MS.