The structural basis of alpha-glucan recognition by a family 41 carbohydrate-binding module from Thermotoga maritima

Starch recognition by carbohydrate-binding modules (CBMs) is important for the activity of starch-degrading enzymes. The N-terminal family 41 CBM, TmCBM41 (from pullulanase PulA secreted by Thermotoga maritima) was shown to have alpha-glucan binding activity with specificity for alpha-1,4-glucans but was able to tolerate the alpha-1,6-linkages found roughly every three or four glucose units in pullulan. Using X-ray crystallography, the structures were solved for TmCBM41 in an uncomplexed form and in complex with maltotetraose and 6(3)-alpha-D-glucosyl-maltotriose (GM3). Ligand binding was facilitated by stacking interactions between the alpha-faces of the glucose residues and two tryptophan side-chains in the two main subsites of the carbohydrate-binding site. Overall, this mode of starch binding is quite well conserved by other starch-binding modules. The structure in complex with GM3 revealed a third binding subsite with the flexibility to accommodate an alpha-1,4- or an alpha-1,6-linked glucose.     

Crystal Structure of a Full-Length Autotransporter

The autotransporter (AT) secretion mechanism is the most common mechanism for the secretion of virulence factors across the outer membrane (OM) from pathogenic Gram-negative bacteria. In addition, ATs have attracted biotechnological and biomedical interest for protein display on bacterial cell surfaces. Despite their importance, the mechanism by which passenger domains of ATs pass the OM is still unclear. The classical view is that the β-barrel domain provides the conduit through which the unfolded passenger moves, with the energy provided by vectorial folding of the β-strand-rich passenger on the extracellular side of the OM. We present here the first structure of a full-length AT, the esterase EstA from Pseudomonas aeruginosa, at a resolution of 2.5 Å. EstA has a relatively narrow, 12-stranded β-barrel that is covalently attached to the passenger domain via a long, curved helix that occupies the lumen of the β-barrel. The passenger has a structure that is dramatically different from that of other known passengers, with a globular fold that is dominated by α-helices and loops. The arrangement of secondary-structure elements suggests that the passenger can fold sequentially, providing the driving force for passenger translocation. The esterase active-site residues are located at the apical surface of the passenger, at the entrance of a large hydrophobic pocket that contains a bound detergent molecule that likely mimics substrate. The EstA structure provides insight into AT mechanism and will facilitate the design of fusion proteins for cell surface display.     

退休教师骨密度的超声研究

目的：了解老年教师骨质疏松病的情况,用超声骨密度仪对813例老年教师做跟骨超声振幅衰减值研究。方法：超声测量是一种新技术,利用不同密度的物体衰减值不同及穿过不同密度物体声速不同等特性对跟骨进行成像,并计算最具说明性区域的一系列数据。参数中BUA值为最主要的参数,三根等距离曲线的形态和宽度能直观分析出年龄与骨量变化的关系。结果：本文男性平均BUA为63.87dB/MHz,得出回归公式：体重=189.1417 0.2062BUA-0.0224SOS。女性平均BUA为58.9dB/MHz,各年龄分组的BUA值与参考值很接近,统计学显示无意义。结论：调查发现男性和女性的骨质不同,男性BUA骨峰值比女性高。BUA值与其体重有密切关系。在60岁以后（或绝经1-5年后）年龄增长与BUA值无明显关系,说明随着年龄的增加骨量减少很小或不减少。利用这种关系,假如BUA值突然下降则提示近期发生骨质疏松。     

Electron Density Sharpening as a General Technique in Crystallographic Studies

Sharpening is a powerful method to restore the details from blurred electron density in crystals with high overall temperature factors (B-factors). This valuable technique is currently not optimally used because of the uncertainty in the scope of its application and ambiguities in practice. We performed an analysis of ~ 2000 crystal data sets deposited in the Protein Data Bank and show that sharpening improves the electron density map in many cases across all resolution ranges, often with dramatic enhancement for mid- and low-resolution structures. It is effective when used with either experimental or model phases without introducing additional bias. Our tests also provide a practical guide for optimal sharpening. We further show that anisotropic diffraction correction improves electron density in many cases but should be used with caution. Our study demonstrates that a routine practice of electron density sharpening may have a broad impact on the outcomes of structural biology studies.     

The Four Canonical TPR Subunits of Human APC/C Form Related Homo-Dimeric Structures and Stack in Parallel to Form a TPR Suprahelix

The anaphase-promoting complex or cyclosome (APC/C) is a large E3 RING-cullin ubiquitin ligase composed of between 14 and 15 individual proteins. A striking feature of the APC/C is that only four proteins are involved in directly recognizing target proteins and catalyzing the assembly of a polyubiquitin chain. All other subunits, which account for > 80% of the mass of the APC/C, provide scaffolding functions. A major proportion of these scaffolding subunits are structurally related. In metazoans, there are four canonical tetratricopeptide repeat (TPR) proteins that form homo-dimers (Apc3/Cdc27, Apc6/Cdc16, Apc7 and Apc8/Cdc23). Here, we describe the crystal structure of the N-terminal homo-dimerization domain of Schizosaccharomyces pombe Cdc23 (Cdc23^Nterm). Cdc23^Nterm is composed of seven contiguous TPR motifs that self-associate through a related mechanism to those of Cdc16 and Cdc27. Using the Cdc23^Nterm structure, we generated a model of full-length Cdc23. The resultant “V”-shaped molecule docks into the Cdc23-assigned density of the human APC/C structure determined using negative stain electron microscopy (EM). Based on sequence conservation, we propose that Apc7 forms a homo-dimeric structure equivalent to those of Cdc16, Cdc23 and Cdc27. The model is consistent with the Apc7-assigned density of the human APC/C EM structure. The four canonical homo-dimeric TPR proteins of human APC/C stack in parallel on one side of the complex. Remarkably, the uniform relative packing of neighboring TPR proteins generates a novel left-handed suprahelical TPR assembly. This finding has implications for understanding the assembly of other TPR-containing multimeric complexes.     

柠条抗旱的生理生态与土壤水分关系的研究

采用LI－1600气孔计和压力室测试了风沙土上生长的柠条的SPAC系统的水分关系和P－V水分特征曲线,结果表明：当沙土含水率（W）的水分特征曲线的回归方程为：ψs=-40．32196×0．427905W,沙土平均含水率大于45％,柠条固沙林生长正常,叶水势高。沙土含水率小于4．5％时,柠条生长速度显著下降,柠条清晨叶水势（ψm）与沙土含水率（W）的回归方程为：ψm=－0．35926·e6．31144／W,风沙土上生长的柠条的“经济水阈”大约在4．5％左右,“生命水阈”大约在3．5％左右。     

Proliferation and apoptosis in subcutaneous adipose tissue of lactating cows with different genetic merit for milk yield

The aim of this study was to investigate the adipocyte size and fate in subcutaneous fat (scAT) of cows diverging for genetic merit at mid lactation stage, when anabolic activity increases and animals are in a state of positive energy balance. Twenty mid lactation cows (180 ± 20 days in milk) grouped according to the Estimated Breeding Values (EBV) for milk yield in plus (EBVp) and minus (EBVm) variants were selected. Average of adipocytes area, proliferation and apoptotic labelling index as well as DLK-1 expression, a marker of pre-adipocytes, were immunohistochemically evaluated in scAT biopsies. In EBVp cows, the BCS was lower (P < 0.01) whereas milk yield, protein, fat yield (P < 0.001) and plasma free fatty acid concentration (P < 0.05) were higher. The scAT of EBVp cows showed a significantly (P < 0.001) higher frequency between 500 and 3000 μm² classes in comparison to EBVm cows, that showed a significantly (P < 0.01) higher apoptotic labeling index. The immunohistochemical reaction showed DLK-1 positivity in scAT of EBVp cows. Taking together, the data indicate a link between milk yield genetic merit of cows, scAT morphology and function, suggesting greater dynamics and metabolic flexibility in EBVp cows.     

Mean Figures and Mean Shapes Applied to Biological Figure and Shape Distributions in the Plane

Fundamental propositions for mean figures and mean shapes in the plane are proved by using complex numbers. A test of equality of two figure or shape distributions is introduced and applied to data from larvae of mussels and from saurian heads.     

Associations between genetic merit for milk production and animal parameters and the fertility performance of dairy cows

Relationships between genetic merit for milk production and animal parameters and various parameters of reproductive performance were examined using multilevel binary response analysis in a study of 19 dairy herds for three successive years, representing approximately 2500 cows per year. The proportion of cows intended for rebreeding that were back in-calf again within 100 days of calving (ICR-100) and the proportion of cows that reappeared again with 365 (RR-365) and 400 days (RR-400) of a previous calving were considered in addition to the traditional measures of reproductive performance. Each 100-kg increase in genetic merit for milk yield was associated with an increased interval to first service (IFS) and calving index (CI) of 1.4 ( P < 0.001) and 1.8 days ( P < 0.001), respectively, a 0.5% increase ( P < 0.05) in calving rate to first insemination (CR-1) and 0.8% increase in RR-400. Each £10 increase in £PIN (the economically weighted yield selection index used in the UK that takes account of butterfat and protein yields) was associated with an increased IFS and CI of 1.5 ( P < 0.001) and 3.0 days ( P < 0.001), respectively. Cows with increased genetic merit for milk yield and £PIN were more likely to re-calve (RR-overall; P < 0.001). Each 1000-kg increase in 305-day milk yield was associated with an increased IFS and CI of 3.2 ( P < 0.001) and 7.8 days ( P < 0.001), respectively, and a 13.6 ( P < 0.001), 22.4 ( P < 0.001), 19.9 ( P < 0.001) and 19.0% ( P < 0.001) decrease in CR-1, ICR-100, RR-365 and RR-400, respectively. A 10-kg increase in maximum yield was associated with a 6.6-day increase in CI ( P < 0.001) and a 14.9 ( P < 0.001), 18.3 ( P < 0.001), 9.6 ( P < 0.05) and 14.2% ( P < 0.001) decrease in CR-1, ICR-100, RR-365 and RR-400, respectively. Fertility performance was also associated with season of calving, lactation number and dystocia score. Level of production had a larger effect on fertility performance than genetic merit for milk production suggesting that infertility at an individual cow level is more likely to be associated with increased production and an inability to meet the nutritional requirements of the cow.     

Optimization of dose and image quality of paediatric cardiac catheterization procedure

The purpose of this study is to quantify the quality of the available imaging modes for various iodine-based contrast agent concentration in paediatric cardiology. The figure of merit (FOM) was defined as the squared signal to noise ratio divided by a patient dose related parameter. An in house constructed phantom simulated a series of vessel segments with iodine concentrations from 10% or 30 mg/cc to 16% or 48 mg/cc of iodine in a blood plasma solution, all within the dimensional constraints of a paediatric patient. The phantom also used test inserts of tin (Sn). Measurements of Entrance Surface Air Kerma (ESAK) and exit dose rate were performed along with calculations of the signal-to-noise ratio (SNR) of all the objects. A first result showed that it was favourable to employ low dose fluoroscopy mode and lower frame rate modes in cine acquisition if dynamic information is not critical. Normal fluoroscopy dose mode provided a considerably higher dose level (in comparison to low dose mode) with only a slight improvement in SNR. Higher frame rate cine modes should be used however when the clinical situation dictates so. This work also found that tin should not be intended as iodine replacement material for research purposes due to the mismatching SNR, particularly on small vessel sizes.