Sequential deposition of plant glycoproteins and polysaccharides at the host-parasite interface ofUromyces vignae andVigna sinensis

Summary Monokaryotic haustoria (M-haustoria) ofUromyces vignae inVigna sinensis cells are surrounded by an extrahaustorial matrix (ema) and the invaginated host plasmalemma, the extrahaustorial membrane (ehrn). The ema was characterized with antibodies against components of the plant cell wall; the ema contained hydroxyproline-rich glycoproteins and arabinogalactans/arabinogalactan proteins, both at a higher concentration close to the ehm. Haustoria with large vacuoles had the ema encased by additional layers. An electron-translucent inner layer deposited on top of the ema contained arabinogalactans/arabinogalactan proteins, hydroxyproline-rich glycoproteins, and callose. The inner layer was surrounded by an electron-translucent middle layer with numerous dark inclusions, rich in pectin and fucose bound to xyloglucans. Finally, a more electron-dense outer layer containing arabinogalactans/arabinogalactan proteins and hydroxyproline-rich glycoproteins encased the whole structure. These polysaccharides, with the exception of callose and un-esterified pectin, were also found in the plant Golgi apparatus. The polysaccharides were synthesized in the trans Golgi cisternae and secreted into the host-parasite interface. The secretory events seem to be coupled to endocytosis since numerous coated pits were found on the ehm too. The pits were elongated, sometimes formed tubules and the coat reacted with an antibody against plant clathrin. Our results suggest intensive membrane recycling around haustoria, together with the secretion of cell wall material, which in the case of more or less vacuolated haustoria seems to be responsible for encasementAbbreviations AG/AGP arabinogalactans and arabinogalactan proteins - BSA bovine serum albumin - ehm extrahaustorial membrane - ema extrahaustorial matrix - HRGP_2b hydroxyproline rich glycoproteins - M-haustorium monokaryotic haustorium - TBS tris buffered saline     

Restoration prospects of abandoned species‐rich sandy grassland in Hungary

Abstract. Secondary succession and seed bank formation was studied in a formerly grazed, abandoned, eastern Hungarian sandy steppe‐meadow (Pulsatillo‐Festucetum). The vegetation was sampled at different elevations of a sand dune which became partly invaded by the tree Robinia pseudo‐acacia ca. 10 yr ago. Pre‐abandonment vegetation records were used as historic references. Though composition of the non‐invaded grassland only changed moderately, dominance of tall grasses (Elymus hispidus, Poa angustifolia) increased significantly at the cost of annuals and low stature perennials. In the stand invaded by Robinia most grassland species were lost and replaced by nitrophytes. Vertical position influenced species abundance, but affected the composition only moderately. Fine‐scale zonation of the vegetation also changed with time. Species richness of the above‐ground vegetation and the seed density of soil samples at the lower elevation were slightly greater than at the higher sites. Seed banks of sensitive grassland specialists (e.g. Pulsatilla pratensis subsp. hungarica) disappeared during grass encroachment. Following extinction from above‐ground vegetation, restoration must rely on dispersal from adjacent areas. In contrast, several annuals and perennials, which survived this degradation stage in the above‐ground vegetation, possessed seed banks. Many of these species became extinct from the vegetation during the Robinia invasion but left viable persistent seeds. This fact is promising for restoration of the Potentillo‐Festucetum sandy pasture. Competitive weedy species and sprouting Robinia can, however, limit seedling establishment.     

沉水植物菹草在低温条件下对重金属Cu、Pb、Zn的吸附和富集

在10℃的较低温度条件下,研究了冬春季节生长旺盛的沉水植物菹草(Potamogeton crispus L.)对重金属离子Cu²⁺,Pb²⁺,Zn²⁺的生物吸附特征及解吸情况,对不同初始浓度重金属水体中的重金属离子去除率情况,以及在此过程中菹草各器官(叶、茎、根茎、根)对重金属离子的富集情况。结果表明,菹草对Cu²⁺,Zn²⁺的吸附在20 min内达到平衡,对Pb²⁺的吸附在50 min内达到平衡,吸附动力学结果符合伪二级动力学方程,决定系数分别达1,1,0.997 8。Freundlich等温线可较好地拟合菹草吸附Cu²⁺,Pb²⁺,Zn²⁺的过程,Cu²⁺,Pb²⁺,Zn²⁺的吸附容量分别达到66.900 6,26.543 0,30.371 8 mg·L^-1。以去离子水作洗脱剂,解吸液中3种重金属离子浓度均低于仪器检出限(0.01 mg·L^-1),解吸程度微弱。投放菹草后,随着初始处理浓度的升高,水体Cu²⁺的去除率先降低后升高,Pb²⁺的去除率的变化趋势与Cu²⁺类似。Zn²⁺去除率则随水体Zn²⁺初始浓度的升高而逐渐升高。菹草各器官对水体3种重金属离子的富集能力不同,排序为Cu²⁺＞Zn²⁺＞Pb²⁺。不同器官对同一种重金属离子的富集量差异显著,叶是富集重金属离子的主要器官。水体重金属离子的初始浓度会影响菹草各器官富集重金属离子的能力,一般随水体重金属初始浓度升高,菹草各器官的重金属离子富集量虽有不同程度的增加但富集系数持续减小。     

Order of steps in the cytochrome C folding pathway: evidence for a sequential stabilization mechanism

Previous work used hydrogen exchange (HX) experiments in kinetic and equilibrium modes to study the reversible unfolding and refolding of cytochrome c (Cyt c) under native conditions. Accumulated results now show that Cyt c is composed of five individually cooperative folding units, called foldons, which unfold and refold as concerted units in a stepwise pathway sequence. The first three steps of the folding pathway are linear and sequential. The ordering of the last two steps has been unclear because the fast HX of the amino acid residues in these foldons has made measurement difficult. New HX experiments done under slower exchange conditions show that the final two foldons do not unfold and refold in an obligatory sequence. They unfold separately and neither unfolding obligately contains the other, as indicated by their similar unfolding surface exposure and the specific effects of destabilizing and stabilizing mutations, pH change, and oxidation state. These results taken together support a sequential stabilization mechanism in which folding occurs in the native context with prior native-like structure serving to template the stepwise formation of subsequent native-like foldon units. Where the native structure of Cyt c requires sequential folding, in the first three steps, this is found. Where structural determination is ambiguous, in the final two steps, alternative parallel folding is found.     

Small proteins fold through transition states with native-like topologies

The folding pathway of common-type acyl phosphatase (ctAcP) is characterized using psi-analysis, which identifies specific chain-chain contacts using bi-histidine (biHis) metal-ion binding sites. In the transition state ensemble (TSE), the majority of the protein is structured with a near-native topology, only lacking one beta-strand and an alpha-helix. psi-Values are zero or unity for all sites except one at the amino terminus of helix H2. This fractional psi-value remains unchanged when three metal ions of differing coordination geometries are used, indicating this end of the helix experiences microscopic heterogeneity through fraying in the TSE. Ubiquitin, the other globular protein characterized using psi-analysis, also exhibits a single consensus TSE structure. Hence, the TSE of both proteins have converged to a single configuration, albeit one that contains some fraying at the periphery. Models of the TSE of both proteins are created using all-atom Langevin dynamics simulations using distance constraints derived from the experimental psi-values. For both proteins, the relative contact order of the TS models is approximately 80% of the native value. This shared value viewed in the context of the known correlation between contact order and folding rates, suggests that other proteins will have a similarly high fraction of the native contact order. This constraint greatly limits the range of possible configurations at the rate-limiting step.     

Carbohydrate derived‐pseudo‐lignin can retard cellulose biological conversion

Dilute acid as well as water only (hydrothermal) pretreatments often lead to a significant hemicellulose loss to soluble furans and insoluble degradation products, collectively termed as chars and/or pseudo‐lignin. In order to understand the factors contributing to reducing sugar yields from pretreated biomass and the possible influence of hemicellulose derived pseudo‐lignin on cellulose conversion at the moderate to low enzyme loadings necessary for favorable economics, dilute acid pretreatment of Avicel cellulose alone and mixed with beechwood xylan or xylose was performed at various severities. Following pretreatment, the solids were enzymatically hydrolyzed and characterized for chemical composition and physical properties by NMR, FT‐IR, and SEM imaging. It was found that hemicelluloses (xylan) derived‐pseudo‐lignin was formed at even moderate severities and that these insoluble degradation products can significantly retard cellulose hydrolysis. Furthermore, although low severity (CSF ～ 1.94) dilute acid pretreatment of a xylan–Avicel mixture hydrolyzed most of the xylan (98%) and produced negligible amounts of pseudo‐lignin, enzymatic conversion of cellulose dropped significantly (>25%) compared to cellulose pretreated alone at the same conditions. The drop in cellulose conversion was higher than realized for cellulase inhibition by xylooligomers reported previously. Plausible mechanisms are discussed to explain the observed reductions in cellulose conversions. Biotechnol. Bioeng. 2013; 110: 737–753. © 2012 Wiley Periodicals, Inc.     

Preparation and optimization of tablets containing a self-nano-emulsifying drug delivery system loaded with rosuvastatin

Background: Rosuvastatin (ROS) calcium is the latest synthetic drug in the statin group that has an anti-hyperlipidemic activity. It is available as tablets, and its poor aqueous solubility, slow dissolution rate and low-absorption extent result in less than 20% bioavailability and about 80% being excreted unchanged in the feces without absorption.

Objective: To utilize nanotechnology to reformulate ROS as a self-nano-emulsifying drug delivery system (SNEDDS), and utilizing design optimization to fabricate the SNEDDS as a tablet.

Methods: The solubility of ROS in different oils, surfactants and co-surfactants was tested. Pseudo-ternary phase diagrams were developed and various SNEDDS formulations were prepared and evaluated regarding globule size, self-emulsification, viscosity and transmittance. The optimized system was examined using transmission electron microscopy. The self-nano-emulsifying tablets were prepared using two types of nano-silica and different percentages of Avicel as a binder and Ac-Di-Sol as a disintegrant. The prepared tablets were evaluated for their physicochemical properties. Bioavailability in human volunteers was assessed.

Results: A SNEDDS system was successfully developed with a droplet size range of 15?nm and a composition of 10% Labrafac, 80% Cremophore RH40 and 10% Propylene glycol. The optimized tablet formula contained: hydrophilic nano-silica, 3% Ac-Di-Sol and 30% Avicel. The pharmacokinetic study revealed that the bioavailability was enhanced by more than 2.4-fold compared with the commercially available tablet.

Conclusions: Tablets containing SNEDDS loaded with ROS represent a promising novel formula that has higher gastrointestinal absorption and enhanced systemic bioavailability.     

Marphysa (Eunicidae,polychaete, Annelida) species of the Sanguinea group from Australia,with comments on pseudo‐cryptic species

The genus Marphysa is widely collected for bait by recreational fishermen in Australia, and yet the number of species remains unresolved. A new species is described together with additional information on a previously described species. This study reveals that while species of Marphysa superficially resemble one another, detailed studies on the distribution of different types of chaetae along the body, together with molecular studies, uncover species which may co‐occur. Correct identification is critical for management of these largely intertidal species. This study also highlights the increasing awareness of the presence of pseudo‐cryptic species in polychaetes.