Effect of Flexible Back on Energy Absorption during Landing in Cats： A Biomechanical Investigation

Cats are characterized by their excellent landing ability. During the landing, they extend and bend their flexible backs. This study was undertaken to examine the effect of flexible back on impact attenuation. We collected kinematic and ground reaction force data from cats performing self-initiated jump down at different heights. Based on these measurements, the mechanical energy and elastic back energy were calculated. Further, we derived a beam model to predict back stiffness from the morphology of the vertebral spines. We found that cat could actively modulate the bending level of flexible back and the landing angle at different heights, making some kinetic energy be stored briefly as elastic strain energy in the back. This mechanism allows cat to reduce the kinetic energy dissipated by limbs and improve the efficiency of energy absorption. These results can provide bio- logical inspiration for the design of a flexible spine on a landing robot, and we anticipated their use in the energy absorption equipments for planetary exploration.     

喀斯特地区农村劳动力转移对生态恢复的影响及其区域差异——基于酉阳和沿河的农户调查数据

喀斯特地区的生态恢复对于促进国土空间的生态修复、实现区域生态安全具有重要意义。基于酉阳县和沿河县的383份农户调查数据，从微观的农户尺度运用Tobit模型分析喀斯特地区农村劳动力转移对生态恢复的影响，并关注其对酉阳县和沿河县生态恢复的影响差异。研究结果表明：①研究区劳动力转移现象明显，47.67%的农村劳动力发生了转移；②研究区生态恢复情况总体较好，酉阳县的生态恢复好于沿河县；③农村劳动力转移通过农业劳动力的流失效应和非农收入的替代效应促进了喀斯特地区的生态恢复；④劳动力转移的程度不同，对生态恢复的作用强度也不同。沿河相对滞后的劳动力转移程度，使得其生态恢复稍弱于酉阳。为促进喀斯特地区生态恢复，政府应有序引导农村劳动力向非农就业转移，提高转移农户的非农化程度。     

Functional RelBE-Family Toxin-Antitoxin Pairs Affect Biofilm Maturation and Intestine Colonization in Vibrio cholerae

Toxin–antitoxin (TA) systems are small genetic elements that typically encode a stable toxin and its labile antitoxin. These cognate pairs are abundant in prokaryotes and have been shown to regulate various cellular functions. Vibrio cholerae, a human pathogen that is the causative agent of cholera, harbors at least thirteen TA loci. While functional HigBA, ParDE have been shown to stabilize plasmids and Phd/Doc to mediate cell death in V. cholerae, the function of seven RelBE-family TA systems is not understood. In this study we investigated the function of the RelBE TA systems in V. cholerae physiology and found that six of the seven relBE loci encoded functional toxins in E. coli. Deletion analyses of each relBE locus indicate that RelBE systems are involved in biofilm formation and reactive oxygen species (ROS) resistance. Interestingly, all seven relBE loci are induced under the standard virulence induction conditions and two of the relBE mutants displayed a colonization defect, which was not due to an effect on virulence gene expression. Although further studies are needed to characterize the mechanism of action, our study reveals that RelBE systems are important for V. cholerae physiology.     

H.pylori可塑区tfs3a分泌系统virB2基因功能

目的对virB2基因编码蛋白进行分析,为virB2基因及其编码蛋白功能提供实验依据。方法利用多种生物学软件以及网站对VirB2蛋白的结构和功能进行分析预测,VirB2蛋白序列通过基因推导获得并由生物公司合成,然后通过免疫动物实验制备鼠抗VirB2蛋白多克隆抗体,同时设计进行VirB2蛋白细胞毒试验(MTT法)。结果virB2基因编码蛋白属于疏水性蛋白,为鞭毛样结构,有较强的细胞毒作用。结论对VirB2蛋白的结构和功能进行了分析预测,证明VirB2蛋白在H.pylori相关的致病性特别是引起胃黏膜炎症方面起到一定的作用,能够为研究H.pylori致病机制提供帮助。     

Algorithmic co-optimization of genetic constructs and growth conditions: application to 6-ACA,a potential nylon-6 precursor

Optimizing bio-production involves strain and process improvements performed as discrete steps. However, environment impacts genotype and a strain that is optimal under one set of conditions may not be under different conditions. We present a methodology to simultaneously vary genetic and process factors, so that both can be guided by design of experiments (DOE). Advances in DNA assembly and gene insulation facilitate this approach by accelerating multi-gene pathway construction and the statistical interpretation of screening data. This is applied to a 6-aminocaproic acid (6-ACA) pathway in Escherichia coli consisting of six heterologous enzymes. A 32-member fraction factorial library is designed that simultaneously perturbs expression and media composition. This is compared to a 64-member full factorial library just varying expression (0.64 Mb of DNA assembly). Statistical analysis of the screening data from these libraries leads to different predictions as to whether the expression of enzymes needs to increase or decrease. Therefore, if genotype and media were varied separately this would lead to a suboptimal combination. This is applied to the design of a strain and media composition that increases 6-ACA from 9 to 48 mg/l in a single optimization step. This work introduces a generalizable platform to co-optimize genetic and non-genetic factors.     

肿瘤特异性蛋白B7-H4纳米抗体的筛选和鉴定

羊驼体内存在天然缺少轻链的重链抗体,克隆重链抗体可变区(VHH),即可构建单域抗体(single-domain antibodies,sdAbs),又称纳米抗体(nanobody,Nb)。利用非免疫羊驼噬菌体文库筛选肿瘤特异性蛋白B7-H4的纳米抗体,经过4轮淘选,ELASE鉴定阳性克隆噬菌体,测序获得其DNA序列后体外转录为mRNA,将修饰纯化后的mRNA转染到肿瘤细胞,利用细胞免疫荧光检测mRNA在肿瘤细胞内是否表达,Western印迹进一步验证其表达情况;通过CCK-8法鉴定其对肿瘤细胞的增殖抑制能力;划痕实验鉴定其对肿瘤细胞迁移能力的影响;Transwell法鉴定其对肿瘤细胞的侵袭能力的影响;裸鼠荷瘤模型瘤旁注射mRNA,鉴定其在体内实验对肿瘤组织的作用。结果显示,通过淘选获得1个高亲和性的噬菌体株菌H6;DNA测序并导出的氨基酸序列符合羊驼纳米抗体结构;将其mRNA导入肿瘤细胞,能有效表达出纳米抗体H6;转染H6-mRNA的肿瘤细胞(0.84±0.08)与未转染H6-mRNA的对照组(1.83±0.04)相比,其增殖能力明显受到抑制,P<0.01,其迁移和侵袭能力(78.60±5.36)明显低于空白对照组(197.80±21.04),效果优于B7-H4 mRNA的siRNA(95.40±16.56);在裸鼠乳腺癌模型中能有效抑制肿瘤生长,效果优于紫杉醇和B7-H4 mRNA的siRNA。这说明筛选所得抗B7-H4纳米抗体H6能特异结合B7-H4蛋白并封闭其功能,导致肿瘤细胞的增殖、迁移和侵袭受到抑制。该结果为利用B7-H4作为治疗癌症的靶点提供了实验基础。     

Influenza Virus Reassortment Is Enhanced by Semi-infectious Particles but Can Be Suppressed by Defective Interfering Particles

A high particle to infectivity ratio is a feature common to many RNA viruses, with ~90–99% of particles unable to initiate a productive infection under low multiplicity conditions. A recent publication by Brooke et al. revealed that, for influenza A virus (IAV), a proportion of these seemingly non-infectious particles are in fact semi-infectious. Semi-infectious (SI) particles deliver an incomplete set of viral genes to the cell, and therefore cannot support a full cycle of replication unless complemented through co-infection. In addition to SI particles, IAV populations often contain defective-interfering (DI) particles, which actively interfere with production of infectious progeny. With the aim of understanding the significance to viral evolution of these incomplete particles, we tested the hypothesis that SI and DI particles promote diversification through reassortment. Our approach combined computational simulations with experimental determination of infection, co-infection and reassortment levels following co-inoculation of cultured cells with two distinct influenza A/Panama/2007/99 (H3N2)-based viruses. Computational results predicted enhanced reassortment at a given % infection or multiplicity of infection with increasing semi-infectious particle content. Comparison of experimental data to the model indicated that the likelihood that a given segment is missing varies among the segments and that most particles fail to deliver ≥1 segment. To verify the prediction that SI particles augment reassortment, we performed co-infections using viruses exposed to low dose UV. As expected, the introduction of semi-infectious particles with UV-induced lesions enhanced reassortment. In contrast to SI particles, inclusion of DI particles in modeled virus populations could not account for observed reassortment outcomes. DI particles were furthermore found experimentally to suppress detectable reassortment, relative to that seen with standard virus stocks, most likely by interfering with production of infectious progeny from co-infected cells. These data indicate that semi-infectious particles increase the rate of reassortment and may therefore accelerate adaptive evolution of IAV.     

Inhibition of N-linked glycosylation affects organic cation transport across the brush border membrane of opossum kidney (OK) cells.

Many organic cations are transported across the apical membrane of the proximal tubule by specific saturable mechanisms. The goal of this study was to determine if the transporter for tetraethylammonium (TEA) in the brush border membrane of an established opossum kidney (OK) cell line is glycosylated and to elucidate the function of this glycosylation. The uptake of TEA was determined in OK cell monolayers treated with tunicamycin (TM), a compound that prevents synthesis of the core oligosaccharide precursor molecules. TM exposure significantly decreased the incorporation of [3H]mannose in OK cell proteins and significantly reduced TEA uptake in a time and a concentration dependent manner. No effect of TM exposure on cellular protein synthesis, DNA content, cell viability, or on [3H]proline uptake was observed. The transport of TEA in control cells was characterized by a Km of 26.9 +/- 16.4 microM and a Vmax of 378 +/- 39 pmol/mg of protein/min. TM treatment (1 microgram/ml for 21 h) significantly increased the Km by over 4-fold to 111.5 +/- 18.4 microM while not affecting the Vmax. The apparent KI values of other organic cations known to interact with this transport system were also significantly increased by TM exposure. Estimated KI values of N1-methylnicotinamide, cimetidine, and mepiperphenidol increased by 6-fold, 4-fold, and 2-fold, respectively, after exposure of OK cells to TM. An increased KI for protons was also observed. Additional inhibitors of the N-linked glycosylation pathway, castanospermine, deoxynojirimycin, and deoxymannojirimycin significantly decreased TEA transport, whereas swainsonine had no effect. Our results suggest that the organic cation transporter is glycosylated. The N-linked oligosaccharide side chain appears to be of the hybrid type, and it either directly or indirectly affects the binding site of the transporter for both organic cations and protons. This is the first report describing the importance of glycosylation in the function of the organic cation transporter in the apical membrane of OK cells.