Effects of drought stress and N supply on the growth, biomass partitioning and water-use efficiency of Sophora davidii seedlings

A greenhouse experiment was conducted in order to understand the adaptation responses to different water and N conditions, and further explore if additional N supply could improve the water-use efficiency (WUE) and adaptability of Sophora davidii seedlings under dry conditions. Two-month-old seedlings were subjected to a completely random design with three water (80, 40 and 20% water field capacity (FC)) and three N supply (N0: 0, Nl: 92 and Nh:184 mg N kg⁻¹ soil) regimes. Drought stress dramatically decreased seedlings height, basal diameter, leaf number, leaf area, root length, and biomass production. An increase in below-ground biomass was observed indicating a higher root/shoot ratio (R/S) under drought stress conditions, and drought further decreased relative water content (RWC) and WUE. On the other hand, S. davidii seedlings exhibited strong responses to N supply, but the responses were inconsistent with the various N supply levels. Low N supply (Nl) increased seedlings height, basal diameter, leaf number, leaf area, and biomass production, but decreased root length. In contrast, high N supply (Nh) decreased or had little effect on these growth characteristics. N supply increased leaf percentages, but decreased fine root percentages. In addition, Nl rather than the other two N treatments increased leaf area ratio (LAR), leaf/fine root mass ratio (L/FR), R/S and RWC under severe drought stress (20% FC), even though these parameters could increase with the Nh treatment under well-watered condition (80% FC). Moreover, Nl also increased WUE under three water conditions, but Nh had little effect on WUE under drought stress conditions (40% FC and 20% FC). The results suggested that water and N co-limited the growth of S. davidii seedlings, and the seedlings exhibited great positive responses to Nl in this study. Appropriate or low N supply, therefore, would be recommended to stimulate growth, enhance WUE, alleviate drought stress, and consequently contribute to S. davidii seedling establishment under dry condition, but excess N supply should be avoided.     

Human alpha-synemin interacts directly with vinculin and metavinculin

Synemin is a very large, unique member of the IF (intermediate filament) protein superfamily. Association of synemin with the major IF proteins, desmin and/or vimentin, within muscle cells forms heteropolymeric IFs. We have previously identified interactions of avian synemin with alpha-actinin and vinculin. Avian synemin, however, is expressed as only one form, whereas human synemin is expressed as two major splice variants, namely alpha- and beta-synemins. The larger alpha-synemin contains an additional 312-amino-acid insert (termed SNTIII) located near the end of the long C-terminal tail domain. Whether alpha- and beta-synemins have different cellular functions is unclear. In the present study we show, by in vitro protein-protein interaction assays, that SNTIII interacts directly with both vinculin and metavinculin. Furthermore, SNTIII interacts with vinculin in vivo, and this association is promoted by PtdIns(4,5)P(2). SNTIII also specifically co-localizes with vinculin within focal adhesions when transiently expressed in mammalian cells. In contrast, other regions of synemin show distinct localization patterns in comparison with those of SNTIII, without labelling focal adhesions. Our results indicate that alpha-synemin, but not beta-synemin, interacts with both vinculin and metavinculin, thereby linking the heteropolymeric IFs to adhesion-type junctions, such as the costameres located within human striated muscle cells.     

Comparative proteomics profile of osteoblasts cultured on dissimilar hydroxyapatite biomaterials: an iTRAQ-coupled 2-D LC-MS/MS analysis

Hydroxyapatite (HA) and its derived bioceramic materials have been widely used for skeletal implants and/or bone repair scaffolds. It has been reported that carbon nanotube (CNT) is able to enhance the brittle ceramic matrix without detrimental to the bioactivity. However, interaction between osteoblasts and these bioceramics, as well as the underlying mechanism of osteoblast proliferation on these bioceramic surfaces remain to be determined. Using iTRAQ-coupled 2-D LC-MS/MS analysis, we report the first comparative proteomics profiling of human osteoblast cells cultured on plane HA and CNT reinforced HA, respectively. Cytoskeletal proteins, metabolic enzymes, signaling, and cell growth proteins previous associated with cell adhesion and proliferation were found to be differentially expressed on these two surfaces. The level of these proteins was generally higher in cells adhered to HA surface, indicating a higher level of cellular proliferation in these cells. The significance of these findings was further assessed by Western blot analysis. The differential protein profile in HA and CNT strengthened HA established in our study should be valuable for future design of biocompatible ceramics.     

人SUMO-3基因原核表达及多克隆抗体制备

构建人SUMO-3基因的原核表达载体pET41a(+)-SUMO-3,表达重组GST-SUMO-3融合蛋白,制备人SUMO-3多克隆抗体。试验结果显示,通过PCR方法从重组质粒pEYFP-SUMO-3中克隆到的SUMO-3 N端93个氨基酸的基因序列与NCBI上提供的序列一致,重组质粒pET41a(+)-SUMO-3构建成功;重组pET41a(+)-SUMO-3在E.coli.BL21 (DE3) pLysS中表达GST-SUMO-3融合蛋白,分子量为44.0 kDa,与预期分子量一致;采用亲和层析纯化融合蛋白GST-SUMO-3并免疫家兔,获得人SUMO-3抗体;Western blot 检测显示该抗体可以特异性识别SUMO-3,ELISA检测结果成阳性,抗体效价约为1: 20000。实验结果为进一步研究人SUMO-3及SUMO第二类家族的功能提供了有用工具。     

Mapping of the nuclear localization signals in open reading frame 2 protein from porcine circovirus type 1

Porcine circovirus type 1 （PCV1） contains two major open reading frames encoding the replication-associated proteins and the major structural capsid （Cap） protein. PCV1 Cap has an N-terminus carrying several potential monopartite or bipartite nuclear localization signals （NLS）. The contribution of these partially overlapping motifs to nuclear importing was identified by expression of mutated PCVI Cap versions fused to enhanced green fluorescent protein （EGFP）. The Cterminus truncated PCV1 Cap-EGFP was localized in nuclei of PK-15 cells similar to the wild-type PCV1 Cap-EGFP, whereas truncation of the N-terminus rendered the fusion protein distributed into cytoplasm, indicating that the nuclear import of PCV1 Cap was efficiently mediated by its N-terminal region. Substitutions of basic residues in stretches 9RR- RR12 or the right part of 25RRPYLAHPAFRNRYRWRRK43 resulted in a diffused distribution of the fusion protein in both nuclei and cytoplasm, indicating that the two NLSs were responsible for restricted nuclear targeting of PCV1 Cap.     

椎体成形术的生物力学研究现状和进展

椎体成形术是通过向椎体病损部位注入骨粘合剂,达到缓减疼痛、增强椎体稳定性的目的。由于椎体成形术的治疗原理、远期疗效、并发症均与其生物力学密切相关,其生物力学引起广泛的关注。本文通过椎体强化的生物力学研究方法及椎体强化后的生物力学特性,总结影响椎体强化效果的因素及影响程度,并对椎体强化生物力学研究中存在的问题及研究前景进行展望。     

锌离子存在下EGCG对前列腺癌细胞生长的影响

本文研究了锌离子存在下EGCG对前列腺癌细胞PC-3生长的影响．研究发现Zn^2＋可以增强EGCG抗癌活性,Zn^2＋存在下。EGCG处理后前列腺癌细胞PC-3克隆形成率显著下降。以RT—PCR、免疫组化方法研究Zn^2＋、EGCG对67kD层粘连蛋白受体（67kD Laminin Receptor,67LR）表达调控,结果表明Znn可通过上调67LR的表达,为EGCG提供更多作用的靶位点,增强EGCG对前列腺癌细胞PC-3的毒性作用。MMP-9是肿瘤侵袭转移过程中关键的基质金属蛋白酶。MMP-9活性与癌细胞的转移潜能密切相关。本文研究发现Zn^2＋、EGCG处理可通过抑制MMP-9活性,降低前列腺癌细胞PC-3的迁移率．其中80umol／LEGCG＋80umol／L Zn^2＋处理24h后显著抑制了PC-3细胞的迁移率。     

Identification and functional characterization of an aggregation domain in long myosin light chain kinase

The functions of long smooth muscle myosin light chain kinase (L-MLCK), a molecule with multiple domains, are poorly understood. To examine the existence of further potentially functional domains in this molecule, we analyzed its amino acid sequence with a tango program and found a putative aggregation domain located at the 4Ig domain of the N-terminal extension. To verify its aggregation capability in vitro, expressible truncated L-MLCK variants driven by a cytomegalovirus promoter were transfected into cells. As anticipated, only the overexpression of the 4Ig fragment led to particle formation in Colon26 cells. These particles contained 4Ig polymers and actin. Analysis with detergents demonstrated that the particles shared features in common with aggregates. Thus, we conclude that the 4Ig domain has a potent aggregation ability. To further examine this aggregation domain in vivo, eight transgenic mouse lines expressing the 4Ig domain (4Ig lines) were generated. The results showed that the transgenic mice had typical aggregation in the thigh and diaphragm muscles. Histological examination showed that 7.70 +/- 1.86% of extensor digitorum longus myofibrils displayed aggregates with a 36.44% reduction in myofibril diameter, whereas 65.13 +/- 3.42% of diaphragm myofibrils displayed aggregates and the myofibril diameter was reduced by 43.08%. Electron microscopy examination suggested that the aggregates were deposited at the mitochondria, resulting in structural impairment. As a consequence, the oxygen consumption of mitochondria in the affected muscles was also reduced. Macrophenotypic analysis showed the presence of muscular degeneration characterized by a reduction in force development, faster fatigue, decreased myofibril diameters, and structural alterations. In summary, our study revealed the existence of a novel aggregation domain in L-MLCK and provided a direct link between L-MLCK and aggregation. The possible significance and mechanism underlying the aggregation-based pathological processes mediated by L-MLCK are also discussed.     

Permineralized rhizomes of the Osmundaceae (Filicales): Diversity and tempo-spatial distribution pattern

The family Osmundaceae is among the most primitive ferns of the Filicales, with an extensive fossil record dating back to the Late Paleozoic. Numerous fossil osmundaceous rhizomes have been documented in the geological history. However, the diversity, variation and distribution pattern of permineralized rhizomes remain poorly known. Here we intend to analyze the fossil records with regard to the diversity and distribution pattern of the osmundaceous rhizomes based on available data. To date, about 83 species ascribed to 14 genera of fossil osmundaceous rhizomes have been described worldwide, assigned to two subfamilies, namely, Thamnopteroideae and Osmundoideae. Geologically, two groups (i.e., Thamnopteroideae and Palaeosmunda) have been reported in the Permian. All the Triassic taxa are from the southern hemisphere. Jurassic osmundaceous rhizomes are abundant and widespread throughout the world, most dominant in the southern hemisphere. During the transition of Jurassic to Cretaceous, the diversity of osmundaceous rhizomes declined rapidly. In the Cretaceous, however, the osmundaceous rhizomes from the northern hemisphere surpass those from the southern hemisphere in generic level for the first time. The Cenozoic taxa diversified in the northern hemisphere with the rise of angiosperms. Geographically, the osmundaceous fossil rhizomes have been found in both hemispheres; the major localities include Ural area of the former USSR, Tasmania of Australia, southern Argentina, Antarctica, northern India, central and western part of North America and northern China. We discuss the origin, radiation, and development of the Osmundaceae based on rhizomes, to help further understand the systematic relation and evolutionary history of the family Osmundaceae.     

Receptor-Associated Protein (RAP) Plays a Central Role in Modulating Aβ Deposition in APP/PS1 Transgenic Mice

Background

Receptor associated protein (RAP) functions in the endoplasmic reticulum (ER) to assist in the maturation of several membrane receptor proteins, including low density lipoprotein receptor-related protein (LRP) and lipoprotein receptor 11 (SorLA/LR11). Previous studies in cell and mouse model systems have demonstrated that these proteins play roles in the metabolism of the amyloid precursor protein (APP), including processes involved in the generation, catabolism and deposition of β-amyloid (Aβ) peptides.

Methodology/Principal Findings

Mice transgenic for mutant APPswe and mutant presenilin 1 (PS1dE9) were mated to mice with homozygous deletion of RAP. Unexpectedly, mice that were homozygous null for RAP and transgenic for APPswe/PS1dE9 showed high post-natal mortality, necessitating a shift in focus to examine the levels of amyloid deposition in APPswe/PS1dE9 that were hemizygous null for RAP. Immunoblot analysis confirmed 50% reductions in the levels of RAP with modest reductions in the levels of proteins dependent upon RAP for maturation [LRP trend towards a 20% reduction ; SorLA/LR11 statistically significant 15% reduction (p<0.05)]. Changes in the levels of these proteins in the brains of [APPswe/PS1dE9](+/−)/RAP(+/−) mice correlated with 30–40% increases in amyloid deposition by 9 months of age.

Conclusions/Significance

Partial reductions in the ER chaperone RAP enhance amyloid deposition in the APPswe/PS1dE9 model of Alzheimer amyloidosis. Partial reductions in RAP also affect the maturation of LRP and SorLA/LR11, which are each involved in several different aspects of APP processing and Aβ catabolism. Together, these findings suggest a central role for RAP in Alzheimer amyloidogenesis.