不同土地覆被格局情景下多种生态系统服务的响应与权衡——以雅砻江二滩水利枢纽为例

在二滩水库集水区,按照当地生态政策发展以坡度为指标构建了10种未来土地覆被格局情景,研究生态系统减轻水库泥沙淤积、减轻水库面源污染、产水发电服务及价值对未来覆被格局的响应程度,并兼顾相关产业收益的变化,权衡各种情景格局的服务效益,优选利益相关方福祉提升幅度最大的情景格局。结果表明:1)现有土地覆被格局对减轻水库泥沙淤积功效的发挥不合理,而以退耕还林政策指导区域土地覆被格局变化,对集水区减轻河道和水库泥沙淤积具有较好功效。10种情景下,保沙价值的高低关系同入库泥沙量正好相反,遵循"随地表林地和草地面积比例的增加,保沙价值增加"的规律。随产沙总量的增加,高产沙强度像元呈现从水库周边逐渐向中上游蔓延的趋势。2)退耕还林政策单纯以坡度作为指标,指导土地覆被的转化,对集水区减轻水环境磷素非点源污染功效较差。除了全为未利用地覆被的情景10,余下9类情景模拟的入库磷素量及过滤磷素经济价值的高低关系完全一致,呈农田草地林地的规律。随入库磷素总量的增加,高强度磷素输出像元从雅砻江流域源区和大河湾区,逐渐蔓延扩散至集水区整个水网。3)随着集水区地表林地覆盖面积比例的减少,集水区生态系统总发电净利润值增加,且高净利润像元呈现从冕宁和喜德县的东部逐渐向整个集水区东南部乃至整个下游蔓延的趋势。4)从减轻对环境的负效应以及提升利益相关方福祉的角度,以坡度6°上下划分林地和耕地的情景5和以坡度15°和6°为阈值划分林草农的情景3为除极端的情景外,综合指标提升最高的两种情景。     

Cloning and constitutive expression of His-tagged xylanase GH 11 from Penicillium occitanis Pol6 in Pichia pastoris X33: purification and characterization

High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine residues at the N-terminal and further integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAP) constitutive promoter. Activity assay and SDS-PAGE demonstrate that the His-tagged xylanase was extracellularly expressed in P. pastoris and purified to homogeneity by a simple, one-step purification protocol using immobilized metal affinity chromatography (Ni-NTA resin). The purified PoXyn2 showed a single band on SDS-PAGE with an apparent molecular weight of 30 kDa. The xylanase activity was optimal at pH 3.0 and 50°C. The specific activity measured for Oat Spelt Xylan was 8549.85 U mg(-1). The apparent The K(M) and V(max) values were 8.33±0.7 mg ml(-1)and 58.82±0.9 μmol min(-1) ml(-1), respectively, as measured on Oat Spelt Xylan. This is the first report demonstrating the possibility of mass production of P. occitanis xylanase using P. pastoris.     

Expression patterns of Notch receptors and their ligands in human osteoarthritic and healthy articular cartilage

Notch pathway plays a pivotal role in cell fate determination. There is much interest surrounding its therapeutic potential, in osteoarthritis, but the expression profile of Notch-related molecules, as well as their relation with cartilage pathological parameters, remains unclear. The purpose of our study is to analyze the expression pattern of Notch family members, type II and type I collagen, in normal (healthy) and osteoarthritic human knee cartilage. Osteoarthritic cartilages were obtained from 3 patients undergoing a total knee replacement. Macroscopically normal cartilage was dissected from 3 human knees at the time of autopsy or surgery. Immunohistochemical staining was performed using Notch1,2,3 and 4, Delta, Jagged, type II collagen and type I collagen antibodies. In healthy cartilage, type II collagen was abundantly expressed while type I was absent. This latter increased proportionally to the osteoarthritic grade. Type II collagen expression remained intense in osteoarthritic cartilage. In healthy cartilage as well as in cartilage with minor lesions, Notch family member's proteins were not or just weakly expressed at the surface and in the cells. However, Notch molecules were over-expressed in osteoarthritic cartilage compared to healthy one. This expression pattern was different according to the cartilage zone and the severity of OA. Our data suggest that Notch signaling is activated in osteoarthritic cartilage, compared to healthy cartilage, with a much more abundant expression in the most damaged areas.     

First identification of small-molecule inhibitors of Pontin by combining virtual screening and enzymatic assay

The human protein Pontin, which belongs to the AAA+ (ATPases associated with various cellular activities) family, is overexpressed in several cancers and its silencing in vitro leads to tumour cell growth arrest and apoptosis, making it a good target for cancer therapy. In particular, high levels of expression were found in hepatic tumours for which the therapeutic arsenal is rather limited. The three-dimensional structure of Pontin has been resolved previously, revealing a hexameric assembly with one ADP molecule co-crystallized in each subunit. Using Vina, DrugScore and Xscore, structure-based virtual screening of 2200 commercial molecules was conducted into the ATP-binding site formed by a dimer of Pontin in order to prioritize the best candidates. Complementary to the in silico screening, a versatile and sensitive colorimetric assay was set up to measure the disruption of the ATPase activity of Pontin. This assay allowed the determination of inhibition curves for more than 20 top-scoring compounds, resulting in the identification of four ligands presenting an inhibition constant in the micromolar concentration range. Three of them inhibited tumour cell proliferation. The association of virtual screening and experimental assay thus proved successful for the discovery of the first small-molecule inhibitors of Pontin.     

Alkaline xylanases from Bacillus mojavensis A21: Production and generation of xylooligosaccharides

Medium composition and culture conditions for the xylanases production by Bacillus mojavensis A21 were optimized using two statistical methods: Plackett-Burman design applied to find the key ingredients and conditions for the best yield of enzyme production and Box-Behnken design used to optimize the value of the four significant variables: barley bran, NaCl, agitation, and cultivation time. The optimal conditions for higher production of xylanases were barley bran 18.66g/l, NaCl 1.04g/l, speed of agitation 176rpm and cultivation time 34.08h. Under these conditions, the xylanase experimental yield (7.45U/ml) closely matched the yield predicted by the statistical model (7.23U/ml) with R(2)=0.98. The medium optimization resulted in a 6.83-fold increase in xylanase production compared to that of the initial medium. Best xylanase activity was observed at the temperature of 50°C and at pH 8.0. The enzyme retained more 96% of its activity after 24h at pH ranges from 7.0 to 90.0. The enzyme preserved more 80% of its initial activity after 60min of pre-incubation from 30°C to 60°C. The main hydrolysis products yielded from corncob extracted xylan were xylobiose and xylotriose, suggesting the good potential of strain A21 in xylooligosaccharides production.     

Large gallanes and the PSEPT theory: a theoretical study of Ga n H n+2 clusters (n?=?7–9)

Do alanes Al(n)H(n+2) and gallanes Ga(n)H(n+2) satisfy the polyhedral skeletal electron pair theory (PSEPT)? Taking into account previous work on this question, this paper provides a convincing answer and proposes the reformulation of the (n + 1) electron pairs rule of Wade and Mingos (W-M) for such systems. Following recent studies of tetra-, penta-, hexa-, hepta-, octa-, and nonaalanes as well as valence-isoelectronic/related gallanes, in this paper we present an analysis of the hydrides of aluminum and gallium A(n)H(n+2) (A = Al, Ga and n = 7-9). The aim is still to examine the applicability of PSEPT, especially the W-M rule, to these clusters. Exploration of the total potential energy surfaces (PESs) of hepta-, octa-, and nonagallanes shows that the absolute minima have a nido-like polyhedron arrangement. Unlike the smaller Ga(n)H(n+2) (n = 4, 5, 6), it seems that the size of the cluster largely dictates its preferred geometry. Although none of them have closed (totally triangular) cages, these clusters exhibit significant compactness, comparable to borane double anions, B(n)H(n) (2-), which are the archetypes for the PSEPT theory.     

Muscle activation of the elbow flexor and extensor muscles during self-resistance exercises: comparison of unilateral maximal cocontraction and bilateral self-resistance

Muscle activation during self-resistance exercises was studied in 18 subjects performing (a) maximal unilateral isometric cocontractions of flexor and extensor muscles of the right elbow (UNI); (b) bilateral exercises consisting of maximal isometric extensions of the right elbow against the left elbow flexors (BiExt) and maximal isometric flexion of the right elbow against the left elbow extensors (BiFlex). Force production by the biceps brachii (BB), brachioradialis (BR), and triceps brachii (TB) during UNI, BiFlex, and BiExt were estimated by comparing the integrated surface electromyograms (iEMG) of BB, BR, and TB during UNI, BiExt, and BiFlex with the individual iEMG-force relationship determined from isometric contractions at 30, 60, and 100% maximal voluntary contraction during elbow flexion (MVCflex) or extension (MVCext) against a force transducer. During BiFlex for BB or BR and BiExt for TB, the values (mean ± SE) of BB-iEMG, BR-iEMG, and TB-iEMG were 74.0 ± 4.5, 76.6 ± 5.7, and 84.4 ± 4.5% iEMG at MVC (% iEMGmax). The forces were 86.0 ± 3.7% TB-Forcemax during BiExt, 74.1 ± 3.6% BB-Forcemax and 71.8 ± 4.0% BR-Forcemax during BiFlex. During UNI, BB-iEMG, BR-iEMG, and TB-iEMG were 59.9 ± 4.6, 53.4 ± 4.0, and 66.3 ± 4.7% iEMGmax, respectively. The forces during UNI (70.4 ± 4.0% TB-Forcemax, 60.4 ± 4.3% BB-Forcemax, and 49.2 ± 3.1% BR-Forcemax) were significantly lower than those during bilateral exercises. A 2-way analysis of variance (Muscle × Exercise) indicated that the effects of Muscle and Exercise upon % iEMGmax were significant (p < 0.05; p < 0.001, respectively). In conclusion, bilateral opposition exercises should be more effective in developing strength than cocontraction exercises, which correspond to a moderate activation level even for weak agonist muscle groups.     

Direct and Indirect Relationships Between Specific Leaf Area, Leaf Nitrogen and Leaf Gas Exchange. Effects of Irradiance and Nutrient Supply

We present a series of competing path models relating interspecificpatterns between specific leaf area, leaf nitrogen content,net photosynthesis and stomatal conductance and test these againstdata from 22 species of herbaceous plants grown under controlledconditions with contrasting irradiance and nutrient supply rates.We then compare these results with two previous data sets, onebased on field measures and one based on glasshouse measures,to determine the robustness of the results. Only one model wasable to account for the patterns of direct and indirect effectsbetween the four variables to all data sets. In this model specificleaf area is the forcing variable that directly affects bothleaf nitrogen levels and net photosynthetic rates. Leaf nitrogenthen directly affects net photosynthetic rates which in turnthen affect stomatal conductance to water. Copyright 2001 Annalsof Botany Company Comparative ecology, modelling, path analysis, photosynthesis, plant strategies, SLA, specific leaf area, stomatal conductance     

Involvement of Na(+)/Ca(2+) exchanger in endothelial NO production and endothelium-dependent relaxation

Endothelial nitric oxide (NO) synthase (eNOS) is controlled by Ca(2+)/calmodulin and caveolin-1 in caveolae. It has been recently suggested that Na(+)/Ca(2+) exchanger (NCX), also expressed in endothelial caveolae, is involved in eNOS activation. To investigate the role played by NCX in NO synthesis, we assessed the effects of Na(+) loading (induced by monensin) on rat aortic rings and cultured porcine aortic endothelial cells. Effect of monensin was evaluated by endothelium-dependent relaxation of rat aortic rings in response to acetylcholine and by real-time measurement of NO release from cultured endothelial cells stimulated by A-23187 and bradykinin. Na(+) loading shifted the acetylcholine concentration-response curve to the left. These effects were prevented by pretreatment with the NCX inhibitors benzamil and KB-R7943. Monensin potentiated Ca(2+)-dependent NO release in cultured cells, whereas benzamil and KB-R7943 totally blocked Na(+) loading-induced NO release. These findings confirm the key role of NCX in reverse mode on Ca(2+)-dependent NO production and endothelium-dependent relaxation.