Regulation of cell growth during serum starvation and bacterial survival in macrophages by the bifunctional enzyme SpoT in Helicobacter pylori

In Helicobacter pylori the stringent response is mediated solely by spoT. The spoT gene is known to encode (p)ppGpp synthetase activity and is required for H. pylori survival in the stationary phase. However, neither the hydrolase activity of the H. pylori SpoT protein nor the role of SpoT in the regulation of growth during serum starvation and intracellular survival of H. pylori in macrophages has been determined. In this study, we examined the effects of SpoT on these factors. Our results showed that the H. pylori spoT gene encodes a bifunctional enzyme with both a hydrolase activity and the previously described (p)ppGpp synthetase activity, as determined by introducing the gene into Escherichia coli relA and spoT defective strains. Also, we found that SpoT mediates a serum starvation response, which not only restricts the growth but also maintains the helical morphology of H. pylori. Strikingly, a spoT null mutant was able to grow to a higher density in serum-free medium than the wild-type strain, mimicking the “relaxed” growth phenotype of an E. coli relA mutant during amino acid starvation. Finally, SpoT was found to be important for intracellular survival in macrophages during phagocytosis. The unique role of (p)ppGpp in cell growth during serum starvation, in the stress response, and in the persistence of H. pylori is discussed.     

Synthesis and biological activities of 2-oxocycloalkylsulfonamides

A series of novel 2-oxocycloalkylsulfonamides (4) were synthesized and their structures confirmed by IR, (1)H NMR, and elemental analysis. The bioassay showed that they have fair to excellent fungicidal activities against Botrytis cinerea Pers and Sclerotinia sclerotiorum. Among them, compounds 4A(10), 4A(11), 4A(12), 4B(2), and 4B(3), the EC(50) values of which were 2.12, 3.66, 3.96, 2.38, and 2.43 microg/mL, respectively, displayed excellent fungicidal activity against B. cinerea Pers, and are comparable with commercial fungicide procymidone (the EC(50) value is 2.45 microg/mL). 3D QSAR against B. cinerea Pers was studied, a statistically significant and chemically meaningful CoMFA model was developed and some compounds which have a high predicted activity were forecasted. In addition, the bioassay also showed that the compounds have good inhibitory activities against human tumor cells HL-60, BGC-823, Bel-7402 and KB. It is interesting to point out that the antitumor activities of compounds 4 are in accordance with their fungicidal activity to a great extent: compounds having relatively best antitumor activities (4A(10), 4A(11), 4A(12), and 4B(3)) also displayed excellent fungicidal activity.     

Suppression of telomere-binding protein TPP1 resulted in telomere dysfunction and enhanced radiation sensitivity in telomerase-negative osteosarcoma cell line

Mammalian telomeres are protected by the shelterin complex that contains the six core proteins POT1, TPP1, TIN2, TRF1, TRF2 and RAP1. TPP1, formerly known as TINT1, PTOP, and PIP1, is a key factor that regulates telomerase recruitment and activity. In addition to this, TPP1 is required to mediate the shelterin assembly and stabilize telomere. Previous work has found that TPP1 expression was elevated in radioresistant cells and that overexpression of TPP1 led to radioresistance and telomere lengthening in telomerase-positive cells. However, the exact effects and mechanism of TPP1 on radiosensitivity are yet to be precisely defined in the ALT cells. Here we report on the phenotypes of the conditional deletion of TPP1 from the human osteosarcoma U2OS cells using ALT pathway to extend the telomeres.TPP1 deletion resulted in telomere shortening, increased apoptosis and radiation sensitivity enhancement. Together, our findings show that TPP1 plays a vital role in telomere maintenance and protection and establish an intimate relationship between TPP1, telomere and cellular response to ionizing radiation, but likely has the specific mechanism yet to be defined.     

退耕还林还草过程中的经济补偿问题探讨

经济补偿作一为项政策措施,对保障当前西部开发进程中退耕还林还草工作的顺利展开,发挥着十分重要的作用,然而,作为我国植被生态建设中一个全新的举措。目前经济补偿政策的实施刚刚开始,有关的理论研究尚处于初步的探讨阶段。循着“为什么要补偿？→补偿多少？→如何补偿？”的思路,建立了经济补偿的概念模型,着重探讨了补偿标准的确定以及多层次补偿制度建立等问题,并对目前处于试点示范中的经济补偿政策进行了评述。     

双载体转重链Tyr664Cys和轻链Thr1286Cys突变体BDD-FⅧ基因

用双载体转运凝血Ⅷ因子基因在甲型血友病基因治疗研究中可克服AAV毒载体容量限制,但存在重链分泌低效和链不均衡性问题。为探索重、轻链间二硫键形成对重链分泌的促进作用,该文用双载体转B结构域大部缺失型FⅧ(BDD-FVⅢ)的重链和轻链基因,将重链的Tyr664和轻链Thr1826突变为Cys,研究了HEK293细胞共转基因后的基因表达、分泌至培养上清的重链量和凝血生物活性。用Western blot检测细胞裂解液结果显示,非还原条件下有明显的二硫键交联的重、轻链蛋白;链特异性ELISA定量检测细胞分泌的重链为(125±29)ng/mL,明显高于共转野生型重链和轻链基因细胞的(75±23)ng/mL;Coatest法显示细胞分泌的凝血活性为(0.78±0.29)U/mL,也明显高于共转野生型重链和轻链基因细胞(0.34±0.12)U/mL。结果表明,重、轻链间的二硫键形成可提高双载体转FⅧ基因的功效,为进一步在动物体内转基因提供了实验依据。     

东北虎豹国家公园生态环境质量动态评价及其气候响应

人类活动及气候变化对东北虎豹国家公园生态环境产生深刻的影响,探究该地区生态环境质量的时空演变规律对园区可持续发展具有重要意义。在原有遥感生态指数基础上,选用增强型植被指数代替原绿度指标中的归一化植被指数,构建2001—2020年植被生长季遥感生态指数,探讨园区内核心保护区、虎豹潜在栖息地及主要人口聚集区三大管控区生态环境质量时空变化特征,并基于像元尺度分析其对气候因素的响应。结果表明：(1)近20年虎豹公园生态环境质量呈波动微弱下降趋势,2015年后生态环境质量逐步提升,2020年达到良好水平;(2)园区内各管控区生态环境质量具有明显的空间分异性,东部核心保护区中部及虎豹潜在栖息地中部以南地区生态环境质量较好,主要人口聚集区相对较差。(3)近20年虎豹公园生态环境质量稳定性较差。未来生态环境质量变化趋势为上升的面积占60.86%,表明未来生态环境趋势总体向好,但局部地区仍存在持续恶化态势。(4)虎豹公园生态环境质量主要受降水的影响,与降水呈显著正相关关系,与气温及日照的相关性较弱。     

利用树木年轮重建赣南地区1890年以来 2-3月份温度的变化

采用树木年轮气候学方法,利用江西赣南地区马尾松(Pinus massoniana)的年轮宽度资料,分析了马尾松径向生长与气候要素变化的相关及响应关系,结果表明赣南地区马尾松径向生长与当年2—3月份的平均温度相关性最高。在响应分析的基础上,重建了江西赣南地区1890年以来2—3月份温度的变化历史。重建序列显示在过去119a中研究区存在3个较为明显的冷期(1892—1906年、1918—1922年、1944—1957年)和3个明显的暖期(1909—1917年、1959—1968年和1998—2008年)。     

Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (<i>Eriobotrya japonica</i>)

Loquat (Eriobotrya japonica Lindl.) is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components. Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression. In this study, eight candidate reference genes were selected from our previously published RNA-seq data, and primers for each candidate reference gene were designed and evaluated. The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples, including 12 subgroups of developing or abiotic-stressed tissues. Different combinations of stable reference genes were screened according to a comprehensive rank, which was synthesized from the results of four algorithms, including the geNorm, NormFinder, BestKeeper and ΔCt methods. The screened reference genes were verified by normalizing EjLGA1 in each subgroup. The obtained suitable combinations of reference genes for accurate normalization were GAPDH, EF1α and ACT for floral development; GAPDH, UBCE and ACT for fruit setting; EF1α, GAPDH and eIF2B for fruit ripening; ACT, EF1α and UBCE for leaves under heat stress; eIF2B, UBCE and EF1α for leaves under freezing stress; EF1α, TUA and UBCE for leaves under salt stress; ACT, EF1α and eIF2B for immature pulp under freezing stress; ACT, UBCE and eIF2B for immature seeds under freezing stress; EF1α, eIF2B and UBCE for both immature pulp and seeds under freezing stress; UBCE, TUB and TUA for red-fleshed fruits under cold-storage stress; eIF2B, RPS3 and TUB for white-fleshed fruits under cold-storage stress; and eIF2B, UBCE and RPS3 for both red- and white-fleshed fruits under cold-storage stress. This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat. To our knowledge, this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E. japonica. The use of the three most stable reference genes could increase the reliability of future quantification experiments.