Analysis of CD44-containing lipid rafts: Recruitment of annexin II and stabilization by the actin cytoskeleton.

CD44, the major cell surface receptor for hyaluronic acid (HA), was shown to localize to detergent-resistant cholesterol-rich microdomains, called lipid rafts, in fibroblasts and blood cells. Here, we have investigated the molecular environment of CD44 within the plane of the basolateral membrane of polarized mammary epithelial cells. We show that CD44 partitions into lipid rafts that contain annexin II at their cytoplasmic face. Both CD44 and annexin II were released from these lipid rafts by sequestration of plasma membrane cholesterol. Partition of annexin II and CD44 to the same type of lipid rafts was demonstrated by cross-linking experiments in living cells. First, when CD44 was clustered at the cell surface by anti-CD44 antibodies, annexin II was recruited into the cytoplasmic leaflet of CD44 clusters. Second, the formation of intracellular, submembranous annexin II-p11 aggregates caused by expression of a trans-dominant mutant of annexin II resulted in coclustering of CD44. Moreover, a frequent redirection of actin bundles to these clusters was observed. These basolateral CD44/annexin II-lipid raft complexes were stabilized by addition of GTPgammaS or phalloidin in a semipermeabilized and cholesterol-depleted cell system. The low lateral mobility of CD44 in the plasma membrane, as assessed with fluorescent recovery after photobleaching (FRAP), was dependent on the presence of plasma membrane cholesterol and an intact actin cytoskeleton. Disruption of the actin cytoskeleton dramatically increased the fraction of CD44 which could be recovered from the light detergent-insoluble membrane fraction. Taken together, our data indicate that in mammary epithelial cells the vast majority of CD44 interacts with annexin II in lipid rafts in a cholesterol-dependent manner. These CD44-containing lipid microdomains interact with the underlying actin cytoskeleton.     

Protochlorophyllide forms and energy transfer in dark-grown wheat leaves. Studies by conventional and laser excited fluorescence spectroscopy between 10 K–100 K

The fluorescence properties and role in energy transfer of protochlorophyllide (Pchlide) forms were studied in dark-grown wheat leaves by conventional and laser excited high resolution methods in the 10 K–100 K temperature range. The three major spectral bands, with emission maxima at 633, 657 (of highest intensity) and 670 nm as Bands I, II, and III were analyzed and interpreted as the contributions of six different structural forms. Band I is the envelope of three (0,0) emission bands with maxima at 628, 632 and 642 nm. Laser excitation studies in the range of Band II at 10 K reveal the presence of a spectrally close donor band besides the acceptor, Band II. The intensity in Band III originates mostly from being the vibronic satellite of Band II, but contains also a small (0,0) band with absorption maximum at 674 nm. Excitation spectra show that besides the Pchlides with absorption around 650 nm within Band II, another significant population of Band I with absorption around 640 nm is also coupled by energy transfer to the acceptor of Band II. The spectral difference between the two donor forms indicate different dipolar environments. Upon increasing the temperature, the intensity of Band II and its satellite, Band III decrease, while Band I remains unaffected. Band II shows also a broadening towards the blue side at higher temperatures. Both the quenching of fluorescence and the spectral change was explained by a thermally activated formation of a non-fluorescent intermediate state in the excited state of Pchlide acceptors.     

The Holdridge life zones of the conterminous United States in relation to ecosystem mapping

Aim Our main goals were to develop a map of the life zones for the conterminous United States, based on the Holdridge Life Zone system, as a tool for ecosystem mapping, and to compare the map of Holdridge life zones with other global vegetation classification and mapping efforts. Location The area of interest is the forty-eight contiguous states of the United States. Methods We wrote a PERL program for determining life zones from climatic data and linked it to the image processing workbench (IPW). The inputs were annual precipitation (Pann), biotemperature (T_bio), sea-level biotemperature (T₀bio), and the frost line. The spatial resolution chosen for this study (2.5 arc-minute for classification, 4-km for mapping) was driven by the availability of current state-of-the-art, accurate and reliable precipitation data. We used the Precipitation-elevation Regressions on Independent Slopes Model, or PRISM, output for the contiguous United States downloaded from the Internet. The accepted standard data for air temperature surfaces were obtained from the Vegetation/Ecosystem Modelling and Analysis Project (VEMAP). This data set along with station data obtained from the National Climatic Data Center for the US, were used to develop all temperature surfaces at the same resolution as the Pann. Results The US contains thirty-eight life zones (34% of the world's life zones and 85% of the temperate ones) including one boreal, twelve cool temperate, twenty warm temperate, four subtropical, and one tropical. Seventy-four percent of the US falls in the ‘basal belt’, 18% is montane, 8% is subalpine, 1% is alpine, and < 0.1% is nival. The US ranges from superarid to superhumid, and the humid province is the largest (45% of the US). The most extensive life zone is the warm temperate moist forest, which covers 23% of the country. We compared the Holdridge life zone map with output from the BIOME model, Bailey's ecoregions, Küchler potential vegetation, and land cover, all aggregated to four cover classes. Despite differences in the goals and methods for all these classification systems, there was a very good to excellent agreement among them for forests but poor for grasslands, shrublands, and nonvegetated lands. Main conclusions We consider the life zone approach to have many strengths for ecosystem mapping because it is based on climatic driving factors of ecosystem processes and recognizes ecophysiological responses of plants; it is hierarchical and allows for the use of other mapping criteria at the association and successional levels of analysis; it can be expanded or contracted without losing functional continuity among levels of ecological complexity; it is a relatively simple system based on few empirical data; and it uses objective mapping criteria.     

油菜双单倍体诱导育种技术研究进展

油菜是食用油、优质饲料蛋白的重要来源,杂种优势利用是油菜培育优势性状最重要的手段,且提高亲本的选育效率对优质品种的培育具有积极的推动作用。现有油菜育种技术存在效率低、周期长、盲目性大、应用范围有限等诸多问题,不适于油菜产业快速发展的需求。双单倍体诱导育种技术是近年来新兴的一种快速选育油菜新品种的技术方法。该技术以操作简便、应用范围广、效率高等优势被广泛应用于油菜新品种的选育过程中。从油菜双单倍体诱导技术创新研究的发现、作用表现、诱导机制、作用价值等方面系统地综述了油菜双单倍体诱导技术的研究进展,展望了油菜双单倍体诱导技术的应用前景,以期为未来油菜双单倍体诱导技术以及其他作物诱导系的研究和利用提供参考。     

Local ecological divergence of two closely related stag beetles based on genetic,morphological, and environmental analyses

The process of phenotypic adaptation to the environments is widely recognized. However, comprehensive studies integrating phylogenetic, phenotypic, and ecological approaches to assess this process are scarce. Our study aims to assess whether local adaptation may explain intraspecific differentiation by quantifying multidimensional differences among populations in closely related lucanid species, Platycerus delicatulus and Platycerus kawadai, which are endemic saproxylic beetles in Japan. First, we determined intraspecific analysis units based on nuclear and mitochondrial gene analyses of Platycerus delicatulus and Platycerus kawadai under sympatric and allopatric conditions. Then, we compared differences in morphology and environmental niche between populations (analysis units) within species. We examined the relationship between morphology and environmental niche via geographic distance. P. kawadai was subdivided into the “No introgression” and “Introgression” populations based on mitochondrial COI gene – nuclear ITS region discordance. P. delicatulus was subdivided into “Allopatric” and “Sympatric” populations. Body length differed significantly among the populations of each species. For P. delicatulus, character displacement was suggested. For P. kawadai, the morphological difference was likely caused by geographic distance or genetic divergence rather than environmental differences. The finding showed that the observed mitochondrial–nuclear discordance is likely due to historical mitochondrial introgression following a range of expansion. Our results show that morphological variation among populations of P. delicatulus and P. kawadai reflects an ecological adaptation process based on interspecific interactions, geographic distance, or genetic divergence. Our results will deepen understanding of ecological specialization processes across the distribution and adaptation of species in natural systems.     

Tempo of Degeneration Across Independently Evolved Nonrecombining Regions

Recombination is beneficial over the long term, allowing more effective selection. Despite long-term advantages of recombination, local recombination suppression can evolve and lead to genomic degeneration, in particular on sex chromosomes. Here, we investigated the tempo of degeneration in nonrecombining regions, that is, the function curve for the accumulation of deleterious mutations over time, leveraging on 22 independent events of recombination suppression identified on mating-type chromosomes of anther-smut fungi, including newly identified ones. Using previously available and newly generated high-quality genome assemblies of alternative mating types of 13 Microbotryum species, we estimated degeneration levels in terms of accumulation of nonoptimal codons and nonsynonymous substitutions in nonrecombining regions. We found a reduced frequency of optimal codons in the nonrecombining regions compared with autosomes, that was not due to less frequent GC-biased gene conversion or lower ancestral expression levels compared with recombining regions. The frequency of optimal codons rapidly decreased following recombination suppression and reached an asymptote after ca. 3 Ma. The strength of purifying selection remained virtually constant at d_N/d_S = 0.55, that is, at an intermediate level between purifying selection and neutral evolution. Accordingly, nonsynonymous differences between mating-type chromosomes increased linearly with stratum age, at a rate of 0.015 per My. We thus develop a method for disentangling effects of reduced selection efficacy from GC-biased gene conversion in the evolution of codon usage and we quantify the tempo of degeneration in nonrecombining regions, which is important for our knowledge on genomic evolution and on the maintenance of regions without recombination.     

群体遗传学下动物驯化研究进展

动物驯化是将野生动物改变为能够长期稳定饲养的家养动物的过程。作为新石器时代农业革命的内容,驯化是人类社会文明进步的重要标志之一。由于和人类的密切关系,驯化不仅改变了动物的野生状态,也改变了人类的生活习性和文明进程。动物驯化研究的关键问题包含驯化祖先是谁、驯化所产生的改变及驯化时间地点等。随着高通量基因组技术和对应分析方法的发展,目前研究动物驯化一般基于群体水平,在群体遗传学的框架下研究动物驯化过程中的重要事件。本文总结了群体遗传学下动物驯化研究的相关内容,包括群体动态历史、选择信号、基因交流等,着重介绍了基因选择初始时间和基因交流时间两个新的拓展内容及分析方法,概述了家猪(Sus scrofa f. domestica)、家鸡(Gallus gallus domesticus)、绵羊(Ovis aries)和山羊(Caprine hircus)等几种主要农业动物近期驯化研究的进展,以期为动物驯化研究提供了新的方向和视角。     

完达山东部欧亚水獭种群数量及出现频次影响因素

水獭作为淡水生态系统健康的指示种和旗舰物种,在维持水生生态系统平衡与稳定中发挥着重要作用。然而目前对于完达山东部地区水獭种群数量、分布及其生境选择的影响因素的研究较为匮乏,严重影响了对该物种的野外保护与管理工作。于2021年12月至2022年4月冬季河流封冻期,采用沿河随机样线调查和红外相机监测相结合的方法对完达山东部地区8条主河和23条支流内的欧亚水獭(Lutra lutra)种群数量、分布现状进行了调查,并利用广义可加模型探究水獭出现频次与环境因子的关系。研究结果表明:(1)欧亚水獭在完达山东部水獭种群数量为571-661只,水獭种群密度为沿河(0.5559±0.2898)只/km,呈现中间高,四周低的趋势;(2)环境因子对水獭出现频次影响分析表明,河流深度、距农田距离、距居民区距离和距道路距离是影响水獭出现频次的关键因素。水獭出现频次与河流深度(0-40 cm)呈线性正相关,当河流深度达到40-50 cm时,水獭出现频次最高,之后随着河流深度的增加,出现频次降低;水獭出现频次与距农田距离(0-1.5 km)呈非线性正相关,在距农田距离为1.5-2 km范围内,水獭出现频次最高,之后降低;水獭出现频次与距居民区距离呈线性正相关;水獭出现频次与距道路距离呈非线性关系,当距离>7 km时,二者之间呈现为正相关,反之呈现为负相关。因此,水獭选择栖息地偏向于河流深(40-50 cm),远离农田(1.5-2 km)、居民区和道路(>7 km)的水域。研究为完达山东部欧亚水獭物种保护提供了基础数据和理论依据,建议通过河岸土地覆盖类型的管理、在河岸周围建立森林缓冲区及加强水獭保护宣传力度等措施实现对水獭物种及其栖息地的维持和保护。     

Real‐time droplet size analysis using laser micrometer as a process analytical technology tool for continuous dripping process

Process analysis and monitoring during the manufacturing of the dripping pills are essential. However, research on developing sensor‐based technology or process analytical technology (PAT) tools to analyze and monitor the dripping process is minimal. The purpose of this work is to develop a fast and non‐destructive laser detection system for quantitative visualization of droplets, which involves detecting the size of the droplet and calculating the weight of the dripping pills during the dripping process. Several factors influencing the detection performance of the detection system and the detection system capability for quantitation of the pill weight were explored. The laser detection system accurately detects the weight of the dripping pills with the coefficients of determination (R²) higher than 0.99. It was also robust concerning the variation in critical process parameters and critical material attributes. Furthermore, the laser detection system was successfully applied to the production line of Ginkgo biloba leaf dripping pills to monitor the dripping pills weight. The proposed laser detection system can analyze and monitor the dripping process in dripping pill manufacturing with stable performance, high accuracy, and high efficiency.     

Liraglutide Exerts Protective Effects by Downregulation of PPARγ, ACSL1 and SREBP-1c in Huh7 Cell Culture Models of Non-Alcoholic Steatosis and Drug-Induced Steatosis

(1) Background: With the aging of the population and polypharmacy encountered in the elderly, drug-induced steatosis (DIS) has become frequent cause of non-alcoholic steatosis (NAS). Indeed, NAS and DIS may co-exist, making the ability to distinguish between the entities ever more important. The aim of our study was to study cell culture models of NAS and DIS and determine the effects of liraglutide (LIRA) in those models. (2) Methods: Huh7 cells were treated with oleic acid (OA), or amiodarone (AMD) to establish models of NAS and DIS, respectively. Cells were treated with LIRA and cell viability was assessed by MTT, lipid accumulation by Oil-Red-O staining and triglyceride assay, and intracellular signals involved in hepatosteatosis were quantitated by RT-PCR. (3) Results: After exposure to various OA and AMD concentrations, those that achieved 80% of cells viabilities were used in further experiments to establish NAS and DIS models using 0.5 mM OA and 20 µM AMD, respectively. In both models, LIRA increased cell viability (p < 0.01). Lipid accumulation was increased in both models, with microsteatotic pattern in DIS, and macrosteatotic pattern in NAS which corresponds to greater triglyceride accumulation in latter. LIRA ameliorated these changes (p < 0.001), and downregulated expression of lipogenic ACSL1, PPARγ, and SREBP-1c pathways in the liver (p < 0.01) (4) Conclusions: LIRA ameliorates hepatocyte steatosis in Huh7 cell culture models of NAS and DIS.