Structural basis for GroEL-assisted protein folding from the crystal structure of (GroEL-KMgATP)14 at 2.0A resolution

Nucleotide regulates the affinity of the bacterial chaperonin GroEL for protein substrates. GroEL binds protein substrates with high affinity in the absence of ATP and with low affinity in its presence. We report the crystal structure of (GroEL-KMgATP)(14) refined to 2.0 A resolution in which the ATP triphosphate moiety is directly coordinated by both K(+) and Mg(2+). Upon the binding of KMgATP, we observe previously unnoticed domain rotations and a 102 degrees rotation of the apical domain surface helix I. Two major consequences are a large lateral displacement of, and a dramatic reduction of hydrophobicity in, the apical domain surface. These results provide a basis for the nucleotide-dependent regulation of protein substrate binding and suggest a mechanism for GroEL-assisted protein folding by forced unfolding.     

Tree size and flowering intensity as affected by nitrogen fertilization in non-bearing orange trees grown under Mediterranean conditions

A field experiment was conducted in the South of Portugal with young (0-3 years old) non-bearing ‘Lane Late’ orange trees. Five nitrogen (N) rates were used in a randomised block design with three replicates. The 180 g N tree⁻¹ over three years led to the greatest canopy width (176 cm) and volume (2,697 dm³). The greatest rate applied (720 g N tree⁻¹ in the three years) led to the largest flower yield. Nitrogen concentration in the flowers significantly increased with fertilizer N, and also with the flowering period up to the 23^rd day, declining thereafter. Flower yield was strongly correlated (r = 0.99, p < 0.001) with flower N concentration. Nutrient composition of flowers and of mature leaves from the spring flush was compared. Significant correlations were found for N (r = 0.47, p <0.01), P (r = -0.49, p <0.01), K (r = 0.44, p <0.05) and Ca (r = 0.87, p <0.001), suggesting that flowers can be used as a tool to diagnose the nutritional status of trees. Canonical analysis (with N treatment as dummy-variables) showed strong relationships between canopy width and N, which were greater at the larger rates of fertilizer application, and strong and inverse relationships between K and Mg, also with the greatest N rates.     

Nitrogen vs. phosphorus limitation across an ecotonal gradient in a mangrove forest

Mangrove forests are characterized by distinctive tree-height gradientsthat reflect complex spatial, within-stand differences in environmentalfactors,including nutrient dynamics, salinity, and tidal inundation, across narrowgradients. To determine patterns of nutrient limitation and the effects ofnutrient availability on plant growth and within-stand nutrient dynamics, weused a factorial experiment with three nutrient treatment levels (control, N,P)and three zones along a tree-height gradient (fringe, transition, dwarf) onoffshore islands in Belize. Transects were laid out perpendicular to theshoreline across a mangrove forest from a fringe stand along the seaward edge,through a stand of intermediate height, into a dwarf stand in the interior ofthe island. At three sites, three trees were fertilized per zone for 2yr. Although there was spatial variability in response, growth byR. mangle was generally nitrogen (N) -limited in thefringe zone;phosphorus (P) -limited in the dwarf zone; and, N- and/or P-limited in thetransition zone. Phosphorus-resorption efficiency decreased in all three zones,and N-resorption efficiency increased in the dwarf zone in response to Penrichment. The addition of N had no effect on either P or N resorptionefficiencies. Belowground decomposition was increased by P enrichment in allzones, whereas N enrichment had no effect. This study demonstrated thatessential nutrients are not uniformly distributed within mangrove ecosystems;that soil fertility can switch from conditions of N to P limitation acrossnarrow ecotonal gradients; and, that not all ecological processes respondsimilarly to, or are limited by, the same nutrient.     

小鼠巨核系祖细胞增殖,分化特性的研究

小鼠骨髓细胞经7d培养后进行细胞形态学观察,可见不同发育阶段的巨核细胞及不同大小的巨核细胞集落。通过计数每个集落中的细胞数,可确定相应祖细胞的有丝分裂能力。结果表明,具有不同有丝分裂能力的祖细胞的体外增殖动力学有所不同。祖细胞的数量与其有丝分裂次数呈负相关(r=-0.986)。进行0、1、2和3次有丝分裂的祖细胞的阿糖胞苷自杀率分别为48.9,58.7,48.0和41.2％;放射敏感性的D_O值(Gy)分别为1.71,1.24,1.03和0.77,D_O值的大小与有丝分裂次数呈负相关(r=-0.958)。经3Gy全身照射后CFU-Meg与CFU-GM的恢复动态过程具有不同特点。     

Oxidant and SDS-stable alkaline protease from Bacillus clausii I-52: production and some properties

AIMS: An investigation was carried out on an oxidative and SDS-stable alkaline protease secreted by Bacillus clausii of industrial significance. METHODS AND RESULTS: Maximum enzyme activity was produced when the bacterium was grown in the medium containing (g l-1): soyabean meal, 15; wheat flour, 10; liquid maltose, 25; K2HPO4, 4; Na2HPO4, 1; MgSO4.7H2O, 0.1; Na2CO3, 6. The enzyme has an optimum pH of around 11 and optimum temperature of 60 degrees C. The alkaline protease showed extreme stability towards SDS and oxidizing agents, which retained its activity above 75 and 110% on treatment for 72 h with 5% SDS and 10% H2O2, respectively. Inhibition profile exhibited by phenylmethylsulphonyl fluoride suggested that the protease from B. clausii belongs to the family of serine proteases. CONCLUSIONS: Bacillus clausii produced high levels of an extracellular protease having high stability towards SDS and H2O2. SIGNIFICANCE AND IMPACT OF THE STUDY: The alkaline protease from B. clausii I-52 is significant for an industrial perspective because of its ability to function in broad pH and temperature ranges in addition to its tolerance and stability in presence of an anionic surfactant, like SDS and oxidants like peroxides and perborates. The enzymatic properties of the protease also suggest its suitable application as additive in detergent formulations.     

Autism-Misregulated eIF4G Microexons Control Synaptic Translation and Higher Order Cognitive Functions

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肥沃耕层构建对东北黑土区旱地土壤肥力和玉米产量的影响

有机物料还田是提高土壤肥力、改善土壤结构和增加作物产量的重要农艺措施之一。本研究通过分析有机物料深混还田构建肥沃耕层后土壤的有机质、速效养分含量和玉米产量,明确了黑土区不同土壤类型旱地土壤肥力指标和玉米产量对肥沃耕层构建方式的响应特征,以期为实现东北黑土区旱地保护性利用和农业可持续发展提供科学依据。采用小区试验与大区示范相结合的方式,在黑龙江省、吉林省和辽宁省选取9个生态类型区作为试验点,土壤类型包括黑土(中厚黑土和薄层黑土)、草甸土、黑钙土、白浆土、棕壤、暗棕壤和褐土。每个试验点均设置了玉米秸秆深混构建肥沃耕层(CF_Ⅰ)、秸秆配合有机肥深混构建肥沃耕层(CF_Ⅱ)和无有机物料还田(CK)3个处理。其中,CF_Ⅰ、CF_Ⅱ处理的小区试验和大区示范的秸秆还田量分别为10000 kg·hm^-2和全量还田,CF_Ⅱ处理的有机肥施用量为30000 t·hm^-2;CF_Ⅰ和CF_Ⅱ处理中有机物料的还田深度均为0~35 cm。结果表明: 不同土壤类型旱地的土壤肥力差异较大,不同土层表现为亚耕层土壤肥力小于耕层土壤,其中暗棕壤和白浆土尤为突出;棕壤、褐土耕层和亚耕层的土壤肥力均偏低;黑土和草甸土的质地比较黏重和犁底层较厚。在试验时间为两年以上的5个试验点中,与CK相比,CF_Ⅰ和CF_Ⅱ处理耕层的土壤有机质、碱解氮、速效磷和速效钾含量平均增加1.85 g·kg^-1、20.16 mg·kg^-1、1.56 mg·kg^-1和17.2 mg·kg^-1,亚耕层较耕层增加了2.09 g·kg^-1、12.06 mg·kg^-1、2.18 mg·kg^-1和3.84 mg·kg^-1。与CK相比,CF_Ⅰ处理显著增加了耕作层和亚耕层土壤有机质和速效磷含量,CF_Ⅱ处理显著增加了耕作层和亚耕层的全部土壤肥力指标,说明肥沃耕层构建是提高土壤肥力的重要途径,其中玉米秸秆配施有机肥是快速提升土壤肥力的有效方法。受不同地区水热条件和土壤类型等的影响,不同试验区的玉米产量差异较大;不同处理间差异显著,表现为CF_Ⅱ>CF_Ⅰ>CK,说明肥沃耕层构建方式在不同生态类型区均能有效提高玉米产量。采用玉米秸秆或者玉米秸秆配合有机肥深混的肥沃耕层构建方式能够同步培肥耕层和亚耕层土壤,提高玉米产量。不同生态类型区应根据土壤类型、有机物料来源等采取相应的肥沃耕层构建方式,建议在有机肥源充足的区域,优先采用秸秆配合有机肥深混构建肥沃耕层。     

多囊卵巢综合征肥胖患者血清维生素D、铁蛋白水平、sICAM-1与胰岛素抵抗、糖脂代谢指标的相关性

摘要 目的：探讨多囊卵巢综合征（PCOS）肥胖患者血清维生素D、铁蛋白、可溶性细胞间粘附分子-1（sICAM-1）水平与胰岛素抵抗、糖脂代谢指标的相关性。方法：2018年8月到2021年11月,选择在本院妇科诊治的PCOS患者65例作为研究对象,分为PCOS肥胖组（n=30,体重指数＜28 kg/m²）和PCOS非肥胖组（n=35,体重指数<28 kg/m²）。检测与计算两组清维生素D、铁蛋白、sICAM-1、胰岛素抵抗、糖脂代谢指标并进行相关性分析。结果：两组的血清甲状腺素（T4）、促甲状腺激素（TSH）与泌乳素（PRL）对比差异无统计学意义（P>0.05）,肥胖组的血清促黄体生成素（LH）、促卵泡生成素（FSH）、睾酮（T）水平高于非肥胖组（P<0.05）。肥胖组的血清铁蛋白、sICAM-1水平高于非肥胖组,血清维生素D水平低于非肥胖组（P<0.05）。肥胖组的胰岛素抵抗指数（HOMA-IR）、胰岛素水平（FINS）、甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）较非肥胖组,高密度脂蛋白胆固醇（HDL-C）低于非肥胖组（P<0.05）。在PCOS肥胖患者中,Pearson分析显示血清维生素D、铁蛋白、sICAM-1与胰岛素抵抗、糖脂代谢指标都存在相关性（P<0.05）。结论：PCOS肥胖患者与非肥胖患者的血清维生素D、铁蛋白、sICAM-1、胰岛素抵抗、糖脂代谢指标水平存在差异,血清维生素D、铁蛋白、sICAM-1水平与胰岛素抵抗、糖脂代谢指标存在相关性。     

Alterations in mitochondrial dynamics with age‐related Sirtuin1/Sirtuin3 deficiency impair cardiomyocyte contractility

Sirtuin1 (SIRT1) and Sirtuin3 (SIRT3) protects cardiac function against ischemia/reperfusion (I/R) injury. Mitochondria are critical in response to myocardial I/R injury as disturbance of mitochondrial dynamics contributes to cardiac dysfunction. It is hypothesized that SIRT1 and SIRT3 are critical components to maintaining mitochondria homeostasis especially mitochondrial dynamics to exert cardioprotective actions under I/R stress. The results demonstrated that deficiency of SIRT1 and SIRT3 in aged (24–26 months) mice hearts led to the exacerbated cardiac dysfunction in terms of cardiac systolic dysfunction, cardiomyocytes contractile defection, and abnormal cardiomyocyte calcium flux during I/R stress. Moreover, the deletion of SIRT1 or SIRT3 in young (4–6 months) mice hearts impair cardiomyocyte contractility and shows aging‐like cardiac dysfunction upon I/R stress, indicating the crucial role of SIRT1 and SIRT3 in protecting myocardial contractility from I/R injury. The biochemical and seahorse analysis showed that the deficiency of SIRT1/SIRT3 leads to the inactivation of AMPK and alterations in mitochondrial oxidative phosphorylation (OXPHOS) that causes impaired mitochondrial respiration in response to I/R stress. Furthermore, the remodeling of the mitochondria network goes together with hypoxic stress, and mitochondria undergo the processes of fusion with the increasing elongated branches during hypoxia. The transmission electron microscope data showed that cardiac SIRT1/SIRT3 deficiency in aging alters mitochondrial morphology characterized by the impairment of mitochondria fusion under I/R stress. Thus, the age‐related deficiency of SIRT1/SIRT3 in the heart affects mitochondrial dynamics and respiration function that resulting in the impaired contractile function of cardiomyocytes in response to I/R.