Endocytosis and Exocytosis Events Regulate Vesicle Traffic in Endothelial Cells

We used water-soluble styryl pyridinium dyes that fluoresce at the membrane-water interface to study vesicle traffic in endothelial cells. Cultured endothelial cells derived from bovine and human pulmonary microvessels were incubated in styryl probes, washed to remove dye from the plasmalemmal outer face, and observed by digital fluorescence microscopy. Vesicles that derived from plasmalemma by endocytosis were filled with the styryl dye. These vesicles were distributed throughout the cytosol as numerous particles of heterogeneous diameter and brightness. Vesicle formation was activated 2-fold following addition of extracellular albumin whereas a control protein, immunoglobulin G, had no effect. Dye uptake was abrogated by labeling at low temperatures and inhibitors of phosphoinositide-3-kinase (PI 3-kinase). Tyrosine kinase inhibitors (genistein and herbimycin A) prevented the albumin-induced vesicle formation. Cytochalasin B prevented vesicle redistribution indicating involvement of actin filaments in translocation of endosomes away from sites of vesicle formation. Styryl dye was lost from cells by exocytosis as evident by the disappearance of discrete fluorescent particles. N-ethylmaleimide and botulinum toxin types A and B caused cells to accumulate increased number of vesicles suggesting that exocytosis was regulated by NSF-dependent SNARE mechanism. The results suggest that phosphoinositide metabolism regulates endocytosis in endothelial cells and that extracellular albumin activates endocytosis by a mechanism involving tyrosine phosphorylation, whereas exocytosis is a distinct process regulated by the SNARE machinery. The results support the hypothesis that albumin regulates its internalization and release in vascular endothelial cells via activation of specific endocytic and exocytic pathways.     

Effect of Inositol Deficiency on Cellular Permeability of the Inositol Exacting Yeast

Inositol deficiency caused the abnormalities of permeability of the cell envelope of the inositol exacting yeasts. In the case of Schizosaccharomyces pombe, in which the marked leakage of cellular free-pool fraction was not detected, the uptake activity of glucose or methylglucoside decreased in inositol deficiency, especially in aerobic condition. Investigations on the compositions of lipids and fatty acids showed that the change in fatty acid composition was not so remarkable as that in phosphatides in inositol deficiency. One of the main causes of low transport activity may be due to the change in phosphatides, but not due to that in fatty acids, possibly. Intracellular contents of glucose was not less in inositol deficiency than in sufficiency. These results suggest that inositol deficiency caused the low activity of uptake, which might not be, however, the primary cause of low fermentative activity.

In the case of Saccharomyces cerevisiae Ino^– mutant A–21–20, the similar results about permeability and lipid analyses were obtained in inositol deficiency.     

Using digital image analysis to quantify the architectural parameters of roots grown in thin rhizotrons

Abstract

Roots grown in small chambers or rhizotrons combined with image analysis could be effective for screening traits of root morphology and architecture prior to field experimentation. The objective of this study was to evaluate and compare digital images of root systems with conventional root scans (RSCN) for several sorghum varieties grown in rhizotrons. Plants were grown in duplicate in slim (1.5‐cm thick) rhizotrons angled 15° to the vertical, filled with sandy topsoil and subsoil at field capacity (FC) or 50% available water content (AWC) in a split‐plot design in a glasshouse (16/24°C). Root growth visible on the glass surface was recorded weekly, and roots were washed out 42 and 90 days after sowing for digital imaging of the root systems on pin boards (PBI). The roots were then sub‐sampled to quantify the baseline root parameters from digital RSCN using WinRhizo® software with a flat‐bed scanner. Rhizotron images revealed little of the root system (1–2%) but PBI “recovered” up to 70% of the total root length (TRL) measured by RSCN. A high a priori image contrast, increased image resolution (9.1–12 megapixels) and optimized contrast threshold between roots and background were the key parameters in quantifying the roots from digital images. PBI revealed significant differences in rooting patterns, especially distribution of root length density (RLD) in the profile, which is useful for selecting varieties with improved resource capture. However, the root diameters estimated in the PBI were significantly larger than those measured by RSCN due to lack of contrast, root clustering and overlay.     

数字X线评价低温射频消融术治疗阻塞性睡眠呼吸暂停低通气综合征的 临床疗效

目的:探讨颅咽部数字X线指数对阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患者行低温射频消融手术治疗效果的评价作用。方法:选择2013年10月-2015年10月在我院接受低温射频消融术的OSAHS患者74例作为研究组,另选择同期在我院接受体检的健康志愿者44例作为对照组。分别于手术前后采用X线测量两组研究对象的上齿槽座角(SNA)、下齿槽座角(SNB)、上下齿槽座角(ANB)、颌上角(NA/PA);舌根后方气道前后径(PAS)、软腭后方气道横径(U-MPW)、会厌层气道前后径(V-LPW)、后鼻棘-咽顶点距(PNS-R)、舌骨最上缘至下颌骨垂直距离(H-MP)、软腭长度(SPL)、软腭厚度(SPT)以及舌骨至颈椎前平面距(H-CVP)。结果:研究组患者ANB,NA/PA,PAS,U-MPW,PNS-R及H-CVP均大于对照组,差异具有统计学意义(P0.05);两组SNA、SNB、V-LPW、H-MP、SPL及SPT比较,差异无统计学意义(P0.05);与术前相比,研究组患者术后ANB、PAS、U-MPW及H-CVP均明显改善,差异具有统计学意义(P0.05)。结论:颅咽部数字X线测量指数能够较好的评价OSAHS患者行射频消融术的治疗效果,值得临床参考。     

Long‐term trends in restoration and associated land treatments in the southwestern United States

Restoration treatments, such as revegetation with seeding or invasive species removal, have been applied on U.S. public lands for decades. Temporal trends in these management actions have not been extensively summarized previously, particularly in the southwestern United States where invasive plant species, drought, and fire have altered dryland ecosystems. We assessed long‐term (1940–2010) trends in restoration using approximately 4,000 vegetation treatments conducted on Bureau of Land Management lands across the southwestern United States. We found that since 1940, the proportions of seeding and vegetation/soil manipulation (e.g. vegetation removal or plowing) treatments have declined, while the proportions of prescribed burn and invasive species treatments have increased. Treatments in pinyon‐juniper and big sagebrush communities declined in comparison to treatments in desert scrub, creosote bush, and riparian woodland communities. Restoration‐focused treatment objectives increased relative to resource extraction objectives. Species richness and proportion of native species used in seeding treatments also increased. Inflation‐adjusted costs per area rose 750% for vegetation/soil manipulation, 600% for seeding, and 400% for prescribed burn treatments in the decades from 1981 to 2010. Seeding treatments were implemented in warmer and drier years when compared to the climate conditions of the entire study period and warmer and wetter years relative to several years before and after the treatment. These results suggest that treatments over a 70‐year period on public lands in the southwestern United States are shifting toward restoration practices that are increasingly large, expensive, and related to fire and invasive species control.     

数字化血管三维模型对烧伤患者面部修复术术后皮瓣存活的影响

目的:探讨数字化血管三维模型对烧伤患者面部修复术术后皮瓣存活的影响。方法:选择2012年4月至2017年4月在我院接受组织瓣转移面部修复术的烧伤患者146例,分为对照组(n=64)和研究组(n=82)。对照组术前采用超声多普勒探测患者供区皮瓣的血管位置,走向并标记探测结果,根据多普勒探测结果结合术前设计行皮瓣转移术。观察组术前3~5 d采用CTA技术检查患者供区皮瓣面部受区血管位置、血管走向等,将检查结果通过计算机进行血管三维重建,根据皮瓣血管与受区血管重建模型修订手术方案后行皮瓣转移术。观察和比较手术时间、术后感染发生率、皮瓣断蒂时间、皮瓣血运障碍发生率和术后皮瓣坏死率。结果:研究组手术时间显著短于对照组(P0.01),两组术后感染发生率分别为7.81%和1.21%,研究组明显低于对照组(P0.01)。研究组的皮瓣断蒂时间为17.1±2.5 d,明显短于对照组(21.3±2.8 d,P0.01);对照组皮瓣血运障碍发生率为17.19%,研究组并未发现血运障碍病例。对照组术后皮瓣坏死率为7.81%,而研究组未出现皮瓣坏死病例,存活率为100%,明显高于对照组(P0.05)。结论:术前数字化血管三维模型的建立用于烧伤患者面部修复术可缩短手术时间,提高手术效率,降低术后皮瓣供血障碍的发生率及皮瓣死亡率,有利于患者术后恢复。     

Dynamical stress characterization and energy evaluation of single cardiac myocyte actuating on flexible substrate

Contractility estimation of cardiac myocyte is important for power evaluation of the cell in heart performance. In this paper, we used digital image correlation (DIC) method to obtain dynamic deformation field of the flexible substrate distributively actuated by single cardiac myocyte, which resulted in the dynamic history of traction forces of the cell during contraction-relaxation cycles. Furthermore, the resultant work and power of the single neonatal cardiac myocyte was evaluated to show the energy characteristics of the cells and the responses to the stiffness variation of the substrate. The method provides a useful tool to study behaviors of the cardiac myocytes interacted with the substrates.     

Genotypic variation in seedling root architectural traits and implications for drought adaptation in wheat (Triticum aestivum L.)

Root system characteristics are of fundamental importance to soil exploration and below-ground resource acquisition. Root architectural traits determine the in situ space-filling properties of a root system or root architecture. The growth angle of root axes is a principal component of root system architecture that has been strongly associated with acquisition efficiency in many crop species. The aims of this study were to examine the extent of genotypic variability for the growth angle and number of seminal roots in 27 current Australian and 3 CIMMYT wheat (Triticum aestivum L.) genotypes, and to quantify using fractal analysis the root system architecture of a subset of wheat genotypes contrasting in drought tolerance and seminal root characteristics. The growth angle and number of seminal roots showed significant genotypic variation among the wheat genotypes with values ranging from 36 to 56 (degrees) and 3 to 5 (plant⁻¹), respectively. Cluster analysis of wheat genotypes based on similarity in their seminal root characteristics resulted in four groups. The group composition reflected to some extent the genetic background and environmental adaptation of genotypes. Wheat cultivars grown widely in the Mediterranean environments of southern and western Australia generally had wider growth angle and lower number of seminal axes. In contrast, cultivars with superior performance on deep clay soils in the northern cropping region, such as SeriM82, Baxter, Babax, and Dharwar Dry exhibited a narrower angle of seminal axes. The wheat genotypes also showed significant variation in fractal dimension (D). The D values calculated for the individual segments of each root system suggested that, compared to the standard cultivar Hartog, the drought-tolerant genotypes adapted to the northern region tended to distribute relatively more roots in the soil volume directly underneath the plant. These findings suggest that wheat root system architecture is closely linked to the angle of seminal root axes at the seedling stage. The implications of genotypic variation in the seminal root characteristics and fractal dimension for specific adaptation to drought environment types are discussed with emphasis on the possible exploitation of root architectural traits in breeding for improved wheat cultivars for water-limited environments.     

Disentangling complex fine‐scale ecological patterns by path modelling using GLMM and GIS

Question: How can statistical modelling tools (GLMM) and GIS be used as an aid in understanding complex ecological patterns? This general question was approached by using bryophyte demography data as an example. More specifically, we asked what is the contribution of terrain shape to explaining the performance and fate of plant individuals, controlling for all other known relationships? Location: Norway. Methods: Information on demography was obtained for 140 populations of the perennial clonal bryophyte Hylocomium splendens in Norway spruce forests during an 11‐year period (1992‐2002). Performance (size and branching pattern) was recorded for mature segments and fate was recorded for growing points. Positions of each of the more than 30 000 recorded bryophyte ramets were coupled with (micro‐) topographic characteristics (slope and convexity) derived from fine‐scale digital elevation models in a GIS framework. Carefully planned sequences of generalised linear mixed models (GLMM) were performed to test predictions from a conceptual path model. Results: We demonstrate strong dependence of size on branching, fate and on vertical position in the bryophyte carpet, and an effect of vertical position on branching pattern. Micro‐topography contributed to explaining plant performance by four different mechanisms: (1) a direct effect of slope on the segment's vertical position in the carpet; (2‐3) direct effects of both slope and convexity on fates of individuals via controls on risk of burial; and (4) an indirect effect of convexity on branching pattern via a direct effect on size. No indication of a direct effect of terrain on branching was found. Conclusions: Our study exemplifies the usefulness of GLMM for disentangling complex ecological relationships. Specifically, we recognise micro‐topography as a potentially important factor for plant demography in general and for performance and fate of individuals in particular.     

Land use legacies in post-agricultural forests in the Doupovské Mountains,Czech Republic

Questions: Do differences in previous land use cause long-lasting changes in soil chemistry? Is vegetation composition affected by the previous land use after 50 years of secondary succession? Is the effect of previous land use caused by pre-existing differences in environmental conditions or mediated through changes in soil chemistry? How important is the effect of previous land use in relation to other factors? Location: Doupovské Mountains, Czech Republic. Methods: A stratified random sampling design was used to collect 91 vegetation relevés with accompanying soil samples. The effects of previous land use (arable field, meadow, pasture) on soil pH, organic carbon (C), total nitrogen (N), C:N ratio and available phosphorus were tested by an analysis of covariance. A canonical correspondence analysis and variation partitioning procedure were used to reveal relationships among previous land use, environmental factors and species composition. Results: Organic C, total N and C:N ratio were significantly influenced by previous land use, while available phosphorus and soil pH were not. Previous land use explained a significant part of the variation in species composition and its effects only partly overlapped with the effects of soil chemistry and terrain attributes. However, the species composition of post-agricultural forests was mostly determined by environmental factors not modified by previous land use. Conclusions: Forest communities that originate on abandoned agricultural land are primarily determined by natural environmental conditions. Nevertheless, the type of previous land use also modifies the species assemblages of these forests and needs to be considered as an important determinant of their composition.