Determination of zeranol and its metabolites in bovine muscle and liver by a chemiluminescence enzyme immunoassay: compared to an ultraperformance liquid chromatography tandem mass spectroscopy method

A chemiluminescent enzyme immunoassay (CLEIA) was compared to an ultraperformance liquid chromatography tandem mass spectroscopy (UPLC‐MS/MS) procedure for the analysis of zeranol and its metabolites in bovine tissue samples. Apparent recoveries from fortified samples by both methods were comparable at 0.5–4.0 µg/kg and a significant correlation was obtained. For CLEIA analysis, hapten mimicking the analyte was first synthesized and conjugated with the carrier protein bovine serum albumin as the immunogen to produce monoclonal antibody. The obtained antibody showed extensive cross‐reactivity toward zeranol metabolites (zearalanone). The limit of detection of CLEIA and UPLC‐MS/MS was 0.05 µg/kg and 0.5 µg/kg, respectively. Recoveries of both methods for fortified samples were higher than 75.0% with the coefficient of variation less than 15%. These results indicated that the combination of screening with CLEIA and confirmation with UPLC‐MS/MS for zeranol and its metabolites would be a reliable method for a large number of bovine samples. Copyright © 2013 John Wiley & Sons, Ltd.     

KNOCKDOWN OF THE IONOTROPIC γ‐AMINOBUTYRIC ACID RECEPTOR (GABAR) RDL GENE DECREASES FIPRONIL SUSCEPTIBILITY OF THE SMALL BROWN PLANTHOPPER,Laodelphax striatellus (HEMIPTERA: DELPHACIDAE)

Insect γ‐aminobutyric acid receptors (GABARs) are important molecular targets of cyclodiene and phenylpyrazole insecticides. Previously GABARs encoding rdl (resistant to dieldrin) genes responsible for dieldrin and fipronil resistance were identified in various economically important insect pests. In this study, we cloned the open reading frame cDNA sequence of rdl gene from fipronil‐susceptible and fipronil‐resistant strains of Laodelphax striatellus (Lsrdl). Sequence analysis confirmed the presence of a previously identified resistance‐conferring mutation. Different alternative splicing variants of Lsrdl were noted. Injection of dsLsrdl reduced the mRNA abundance of Lsrdl by 27–82%, and greatly decreased fipronil‐induced mortality of individuals from both susceptible and resistant strains. These data indicate that Lsrdl encodes a functional RDL subunit that mediates susceptibility to fipronil. Additionally, temporal and spatial expression analysis showed that Lsrdl was expressed at higher levels in eggs, fifth‐instar nymphs, and female adults than in third‐instar and fourth‐instar nymphs. Lsrdl was predominantly expressed in the heads of 2‐day‐old female adults. All these results provide useful background knowledge for better understanding of fipronil resistance related ionotropic GABA receptor rdl gene expressed variants and potential functional differences in insects.     

Developmentally regulated sphingolipid synthesis in African trypanosomes

Sphingolipids are essential components of eukaryotic membranes, and many unicellular eukaryotes, including kinetoplastid protozoa, are thought to synthesize exclusively inositol phosphorylceramide (IPC). Here we characterize sphingolipids from Trypanosoma brucei, and a trypanosome sphingolipid synthase gene family (TbSLS1-4) that is orthologous to Leishmania IPC synthase. Procyclic trypanosomes contain IPC, but also sphingomyelin, while surprisingly bloodstream-stage parasites contain sphingomyelin and ethanolamine phosphorylceramide (EPC), but no detectable IPC. In vivo fluorescent ceramide labelling confirmed stage-specific biosynthesis of both sphingomyelin and IPC. Expression of TbSLS4 in Leishmania resulted in production of sphingomyelin and EPC suggesting that the TbSLS gene family has bi-functional synthase activity. RNAi silencing of TbSLS1-4 in bloodstream trypanosomes led to rapid growth arrest and eventual cell death. Ceramide levels were increased more than threefold by silencing suggesting a toxic downstream effect mediated by this potent intracellular messenger. Topology predictions support a revised six-transmembrane domain model for the kinetoplastid sphingolipid synthases consistent with the proposed mammalian sphingomyelin synthase structure. This work reveals novel diversity and regulation in sphingolipid metabolism in this important group of human parasites.     

云南省爬行动物名录和地理区划更新

云南省作为中国生物多样性最高的省份, 其详实的物种本底资料对我国生物多样性研究和保护具有重要意义。本文在前期研究的基础上, 结合实体标本, 汇总编制了云南省现生、原生爬行动物更新名录。截至2021年12月31日, 云南省记录爬行动物25科82属235种, 其中龟鳖目4科12属16种, 有鳞目蜥蜴亚目6科20属72种, 蛇亚目15科50属147种。较《云南两栖爬行动物》确认新增82种, 存疑收录21种, 移除23种。基于先前云南省爬行动物区划和更新后的物种分布信息, 将云南省爬行动物地理分为6个动物地理区, 即滇西北横断山区、滇西山地区、滇南山地区、滇东南山地区、滇中高原区以及滇东北山地区; 其中滇西北横断山区、滇西山地区、滇中高原区和滇东南山地区的范围与先前研究相比有所调整。结合调整后的爬行动物地理区划, 对物种分布、物种特有性、受威胁状况等给出了统计结果。云南省爬行动物特有物种、国内仅见于云南的非特有物种数量较多, 受威胁等级高。建议今后继续加大分类学研究投入, 对滇西北、滇中特有爬行动物分布集中的区域积极开展栖息地保护工作, 同时在最新调整的《国家重点保护野生动物名录》基础上, 定期组织专家研讨, 对《云南省省级重点保护动物名录》提出更新建议。     

Experimental warming shifts coupling of carbon and nitrogen cycles in an alpine meadow

增温对高寒草甸生态系统碳氮循环耦合关系的影响 陆地生态系统碳吸收受土壤氮素可用性的调节。然而,全球变化背景下的不同生态系统组分的碳氮比及其所反映的碳氮循环耦合关系尚不十分清楚。本文运用数据同化的方法,将一个高寒草甸增温试验的14组数据同化到草地生态系统模型中,从而评估了增温如何影响陆地生态系统的碳氮循环耦合关系。研究结果表明,增温提高了土壤氮素的有效性,降低了土壤活性碳库的碳氮比,导致植物对土壤氮的吸收增加。但是由于植物叶片吸收的碳比吸收的氮增加更多,使得叶片中碳氮比增加,而根部的碳输入增加则低于氮的增加,导致根部的碳氮比减少。同时,增温降低了凋落物碳氮比,可能是在土壤高氮有效性的条件下,凋落物氮的固定得到增强;而且增温加速了凋落物的分解。同时增温还增加了慢速土壤有机质的碳氮比,使得该土壤碳库的碳固存潜力增大。由于大多数模型在不同的环境中通常使用相对固定的碳氮比,本研究所发现的气候变暖条件下碳氮比的差异变化可为模型参数化提供一个有效的参考,有利于模型对未来气候变化背景下生态系统碳氮耦合关系响应的预测。     

An apicoplast‐resident folate transporter is essential for sporogony of malaria parasites

Malaria parasites are fast replicating unicellular organisms and require substantial amounts of folate for DNA synthesis. Despite the central role of this critical co‐factor for parasite survival, only little is known about intraparasitic folate trafficking in Plasmodium. Here, we report on the expression, subcellular localisation and function of the parasite's folate transporter 2 (FT2) during life cycle progression in the murine malaria parasite Plasmodium berghei. Using live fluorescence microscopy of genetically engineered parasites, we demonstrate that FT2 localises to the apicoplast. In invasive P. berghei stages, a fraction of FT2 is also observed at the apical end. Upon genetic disruption of FT2, blood and liver infection, gametocyte production and mosquito colonisation remain unaltered. But in the Anopheles vector, FT2‐deficient parasites develop inflated oocysts with unusual pulp formation consisting of numerous single‐membrane vesicles, which ultimately fuse to form large cavities. Ultrastructural analysis suggests that this defect reflects aberrant sporoblast formation caused by abnormal vesicular traffic. Complete sporogony in FT2‐deficient oocysts is very rare, and mutant sporozoites fail to establish hepatocyte infection, resulting in a complete block of parasite transmission. Our findings reveal a previously unrecognised organellar folate transporter that exerts critical roles for pathogen maturation in the arthropod vector.     

Targeting IFN-γ-inducible lysosomal thiol reductase overcomes chemoresistance in AML through regulating the ROS-mediated mitochondrial damage

The persistence of leukemia stem cells (LSCs) is one of the leading causes of chemoresistance in acute myeloid leukemia (AML). To explore the factors important in LSC-mediated resistance, we use mass spectrometry to screen the factors related to LSC chemoresistance and defined IFN-γ-inducible lysosomal thiol reductase (GILT) as a candidate. We found that the GILT expression was upregulated in chemoresistant CD34+ AML cells. Loss of function studies demonstrated that silencing of GILT in AML cells sensitized them to Ara-C treatment both in vitro and in vivo. Further mechanistic findings revealed that the ROS-mediated mitochondrial damage plays a pivotal role in inducing apoptosis of GILT-inhibited AML cells after Ara-C treatment. The inactivation of PI3K/Akt/ nuclear factor erythroid 2-related factor 2 (NRF2) pathway, causing reduced generation of antioxidants such as SOD2 and leading to a shifted ratio of GSH/GSSG to the oxidized form, contributed to the over-physiological oxidative status in the absence of GILT. The prognostic value of GILT was also validated in AML patients. Taken together, our work demonstrated that the inhibition of GILT increases AML chemo-sensitivity through elevating ROS level and induce oxidative mitochondrial damage-mediated apoptosis, and inhibition of the PI3K/Akt/NRF2 pathway enhances the intracellular oxidative state by disrupting redox homeostasis, providing a potentially effective way to overcome chemoresistance of AML.     

胶孢炭疽菌生防放线菌SD-29的鉴定及抗菌活性评价

【背景】胶孢炭疽菌是引起橡胶炭疽病的一种重要病原菌,可导致橡胶树产胶量下降。【目的】从山东青岛一农田土壤中分离出一株胶孢炭疽菌生防放线菌SD-29,并对其进行鉴定及抗菌活性评价。【方法】采用对峙生长法及菌丝生长速率法对菌株SD-29的拮抗活性进行鉴定;利用乙酸乙酯萃取法提取菌株SD-29发酵液粗提物并进行活性评价;根据菌株SD-29的形态特征、生理生化及16S rRNA基因序列进行鉴定。【结果】菌株SD-29对胶孢炭疽菌具有较强的抑制活性,皿内抑制活性达到82.6%。发酵液粗提物对菌丝生长的EC50为13.6μg/mL,100μg/mL的粗提物对胶孢炭疽菌孢子萌发抑制率达到63.16%,其对感炭疽病橡胶叶片的防治效果达到48.96%。根据该菌的形态特征、生理生化特征及16S rRNA基因序列分析鉴定菌株SD-29为Streptomyces yatensis。【结论】菌株SD-29对胶孢炭疽菌有较强的防治效果,具有潜在的应用价值。