Assessment of Mouse Germinal Vesicle Stage Oocyte Quality by Evaluating the Cumulus Layer,Zona Pellucida,and Perivitelline Space

To improve the outcome of assisted reproductive technology (ART) for patients with ovulation problems, it is necessary to retrieve and select germinal vesicle (GV) stage oocytes with high developmental potential. Oocytes with high developmental potential are characterized by their ability to undergo proper maturation, fertilization, and embryo development. In this study, we analyzed morphological traits of GV stage mouse oocytes, including cumulus cell layer thickness, zona pellucida thickness, and perivitelline space width. Then, we assessed the corresponding developmental potential of each of these oocytes and found that it varies across the range measured for each morphological trait. Furthermore, by manipulating these morphological traits in vitro, we were able to determine the influence of morphological variation on oocyte developmental potential. Manually altering the thickness of the cumulus layer showed strong effects on the fertilization and embryo development potentials of oocytes, whereas manipulation of zona pellucida thickness effected the oocyte maturation potential. Our results provide a systematic detailed method for selecting GV stage oocytes based on a morphological assessment approach that would benefit for several downstream ART applications.     

Coarse-grained modeling of mucus barrier properties

We designed a simple coarse-grained model of the glycocalyx layer, or adhesive mucus layer (AML), covered by mucus gel (luminal mucus layer) using a polymer lattice model and stochastic sampling (replica exchange Monte Carlo) for canonical ensemble simulations. We assumed that mucin MUC16 is responsible for the structural properties of the AML. Other mucins that are much smaller in size and less relevant for layer structure formation were not included. We further assumed that the system was in quasi-equilibrium. For systems with surface coverage and concentrations of model mucins mimicking physiological conditions, we determined the equilibrium distribution of inert nanoparticles within the mucus layers using an efficient replica exchange Monte Carlo sampling procedure. The results show that the two mucus layers penetrate each other only marginally, and the bilayer imposes a strong barrier for nanoparticles, with the AML layer playing a crucial role in the mucus barrier.     

长期施肥对农田黑土微生物群落功能多样性的影响

为了研究长期不同施肥对农田黑土微生物群落功能多样性的影响,采用Biolog ECO微平板培养法,对已经连续施肥35年的农业部哈尔滨黑土生态环境重点野外科学观测试验站4种处理(CK、NPK、M、MNPK)的两个土层(0～20、20～40 cm)微生物群落功能多样性进行研究.结果表明: 在0～20 cm土层,有机无机肥(MNPK)配施能够显著提高土壤微生物对碳源的利用能力以及群落代谢功能的丰富度、多样性和优势度,而在20～40 cm土层则低于单施化肥(NPK)处理,两个土层单施化肥均降低土壤微生物群落代谢均匀度.不同施肥处理土壤微生物对6种碳源的利用率在0～20和20～40 cm两个土层间存在差异,而在每个土层的处理间差异显著(P<0.05),但各处理间的变化规律在两个土层中不一致.典范对应分析(CCA)结果表明,各处理土壤微生物群落代谢功能在0～20和20～40 cm土层间存在差异,且在两个土层的处理间也存在差异,而土壤养分含量对各处理土壤微生物群落代谢功能的影响规律在两个土层中较为相似.长期不同施肥会对耕层及以下土壤微生物群落功能多样性产生影响,在20～40 cm土层中单施化肥对微生物群落功能多样性的影响较大.     

长苞铁杉林乔木层优势种群种间关联及尺度效应研究

通过设定不同尺度的样方面积,根据2×2联列表,运用方差分析、联结系数AC、2统计量度量、 Ochiai指数系列技术,测定天宝岩国家级自然保护区长苞铁杉群落乔木层10个优势种群间的总体联结关系、 种对间联结显著性和关联系数。结果表明:长苞铁杉群落乔木层10个优势种群的总体种间关联性均呈正关 联,反映该群落处于稳定的顶极阶段;10个优势种群的种间联结关系中,有26个种对在不同尺度样方设计中 均未达到一般显著水平,种对间的相互独立性较强;样方设计为100 m2时,各项指标上规律性比较明显,它的 研究结果与实际情况相吻合。群落虽处于比较稳定的顶极阶段,但应适当调整群落结构,为长苞铁杉的生长 更新提供良好的生态环境。     

黑龙江省黄鼬冬季毛被分层结构及保温功能

为了解黄鼬毛被的保温机制,选取黑龙江省通河林区黄鼬东北亚种(Mustela sibirica manchurica)冬季皮张,通过观测雄性5个部位和雌性4个部位毛被的分层结构,并结合热物性测试比对,发现了毛被的各层结构具有不同的保温隔热功能且存在部位差异。结果表明:1)毛被由外及里表现出4层结构,各层毛被厚度分别为:(12.7±3.0)mm、(6.0±1.8)mm、(5.5±2.2)mm和(1.4±0.5)mm。2)4层毛被的颜色、结构及保温机制依次为:最外层毛被棕黄色或金黄色,仅由2种类型被毛构成,耐磨损能力较强,保护下层毛被,阻挡冷空气侵入并降低毛被的热量散失;第二层毛被淡黄色,此层开始均由4种类型被毛构成,毛干细度较小,被毛间形成细小空隙,滞留大量静止空气,增强保温功能;第三层毛被灰色或灰白色,4种类型被毛的毛干更细,绒针毛和绒毛弯曲程度更大,使被毛间滞留静止空气的能力更强、更稳定,保温能力更强;最内层毛被为白色,为近毛根处,4种类型被毛均较直,便于热量传递。3)4层毛被的对整体的贡献率分别为:16.11%、27.40%、44.40%和12.09%。4)背面毛被较腹面的颜色深,厚度大,保温能力强;沿吻端至尾基的体轴方向毛被厚度增加,保温性增强。5)雄性毛被较雌性的厚度大,保温能力强。以上,反映了黄鼬冬季多部位毛被由表及里不同的空间布局及各层毛呈现的不同的形态结构,从整体上兼顾保护、保温、散热等多种功能,以适应当地的寒冷环境。     

Reliability of Intra-Retinal Layer Thickness Estimates

Purpose

Measurement of intra-retinal layer thickness using optical coherence tomography (OCT) has become increasingly prominent in multiple sclerosis (MS) research. Nevertheless, the approaches used for determining the mean layer thicknesses vary greatly. Insufficient data exist on the reliability of different thickness estimates, which is crucial for their application in clinical studies. This study addresses this lack by evaluating the repeatability of different thickness estimates.

Methods

Studies that used intra-retinal layer segmentation of macular OCT scans in patients with MS were retrieved from PubMed. To investigate the repeatability of previously applied layer estimation approaches, we generated datasets of repeating measurements of 15 healthy subjects and 13 multiple sclerosis patients using two OCT devices (Cirrus HD-OCT and Spectralis SD-OCT). We calculated each thickness estimate in each repeated session and analyzed repeatability using intra-class correlation coefficients and coefficients of repeatability.

Results

We identified 27 articles, eleven of them used the Spectralis SD-OCT, nine Cirrus HD-OCT, two studies used both devices and two studies applied RTVue-100. Topcon OCT-1000, Stratus OCT and a research device were used in one study each. In the studies that used the Spectralis, ten different thickness estimates were identified, while thickness estimates of the Cirrus OCT were based on two different scan settings. In the simulation dataset, thickness estimates averaging larger areas showed an excellent repeatability for all retinal layers except the outer plexiform layer (OPL).

Conclusions

Given the good reliability, the thickness estimate of the 6mm-diameter area around the fovea should be favored when OCT is used in clinical research. Assessment of the OPL was weak in general and needs further investigation before OPL thickness can be used as a reliable parameter.     

Near-wall micro-PIV reveals a hydrodynamically relevant endothelial surface layer in venules in vivo

High-resolution near-wall fluorescent microparticle image velocimetry (micro-PIV) was used in mouse cremaster muscle venules in vivo to measure velocity profiles in the red cell-depleted plasma layer near the endothelial lining. micro-PIV data of the instantaneous translational speeds and radial positions of fluorescently labeled microspheres (0.47 microm) in an optical section through the midsagittal plane of each vessel were used to determine fluid particle translational speeds. Regression of a linear velocity distribution based on near-wall fluid-particle speeds consistently revealed a negative intercept when extrapolated to the vessel wall. Based on a detailed three-dimensional analysis of the local fluid dynamics, we estimate a mean effective thickness of approximately 0.33 micro m for an impermeable endothelial surface layer or approximately 0.44 micro m assuming the lowest hydraulic resistivity of the layer that is consistent with the observed particle motions. The extent of plasma flow retardation through the layer required to be consistent with our micro-PIV data results in near complete attenuation of fluid shear stress on the endothelial-cell surface. These findings confirm the presence of a hydrodynamically effective endothelial surface layer, and emphasize the need to revise previous concepts of leukocyte adhesion, stress transmission to vascular endothelium, permeability, and mechanotransduction mechanisms.     

天山中段南坡巴伦台地区不同海拔植物群落物种多样性与土壤因子的关系

为探讨天山中段南坡巴伦台植物群落物种多样性随海拔梯度的分布特征及其与土壤环境因子的关系,该研究采用野外调查的方法,在和静县巴伦台地区海拔范围内设置34个样地进行了植物多样性和土壤因子的调查及室内指标的统计分析。结果表明:(1)研究区共调查到30科75属134种植物,草本层为主要优势层。不同海拔高度上土壤理化指标具有异质性,土壤含水量、全盐、有机质、全氮、全钾、有效氮和有效钾差异性显著(P<0.05),其中除全钾以外,其他土壤因子的含量均表现为中海拔大于低、高海拔区域。随海拔的升高,植物群落在低、高海拔段Pielou均匀度指数较高; 灌木层物种Patrick丰富度指数较低; 草本层物种Shannon-Wiener指数、Simpson指数随海拔升高先增加后减小。(2)RDA分析表明,影响植物群落物种多样性的主要环境因子是海拔、土壤含水量、全盐、有机质、全氮和有效氮。海拔作为主导因子,与草本层各物种多样性指数呈正相关,与灌木层各物种多样性指数呈负相关关系; 全盐是抑制植物群落总体物种Simpson指数的主要土壤因子; 氮元素一定程度上限制灌木、半灌木物种的生长。该研究结果表明土壤因子对不同生活型物种多样性的形成具有一定的筛选作用及不同物种对环境变化的适应策略不同。     

家犬嗅球组织学结构的性别和年龄差异

用组织学方法研究家犬嗅球的结构,观察家犬嗅球内结构的性别和年龄差异,依据常规HE染色法及数理统计学原理对家犬嗅球各层宽度,主要细胞的数量进行比较统计学分析,探讨嗅球内部结构的发育过程以及性别差异对雌雄动物嗅觉差异的影响。结果表明：雌雄幼年家犬嗅球内各层结构差异不显著;成年家犬也表现出同样的结果,但是成年动物的僧帽细胞形态、数量差异极显著。分析发现,幼年家犬嗅球各层结构都已比较明显,成年家犬嗅球体积和重量明显增加,各层宽度明显变宽,各层细胞密度显著降低,说明嗅球也处在不断的发育完善过程之中。同时僧帽细胞的差异可能是造成雌雄动物嗅觉差别的原因之一。     

宁夏东部风沙区沙化草地土壤水分和植被的空间特征

土壤水分作为土壤-植被-大气连续体的关键因子,对沙化草地的演化过程具有重要作用。为探讨宁夏东部风沙区沙化草地土壤水分、物种丰富度指数及植被盖度的空间变异及其相互关系,以哈巴湖自然保护区沙化草地为对象,采用样线法自潜在沙化草地至重度沙化草地进行植被调查和土壤取样,通过经典统计学和地统计学分析,得出以下结果：0—100 m各土层土壤水分含量、植被盖度和物种丰富度指数的分布范围分别为0.82%—28.22%、41.00%—93.00%和0.82—2.80,变异系数范围为0.20—0.48,均属于中等变异。各土层土壤水分和物种丰富度指数表现为中等的空间自相关性,植被盖度则表现为强烈的空间自相关性。Kriging插值结果表明,0—100 cm各土层土壤水分和植被盖度的空间插值图呈条带状和斑块状的梯度变化,物种丰富度呈斑块分布,自潜在沙化草地至重度沙化草地,表现为逐渐降低的趋势。相关分析表明,植被盖度与0—40 cm各土层土壤水分呈显著正相关,与40—100 cm各土层土壤水分呈极显著正相关。宁夏东部风沙区沙化草地土壤水分含量总体较低,由于结构因素和随机因素的共同作用,随草地沙化程度的加重,表现为...     

北京小龙门森林生态系统定位站土壤螨类的垂直分布和季节变动

通过对小龙门森林生态系统定位站栎树林中所设立的5个土壤动物采样点,进行每月1次的土壤取样调查(从1993年4月到1994年1月),,对各土层中螨类的垂直分布和季节变动进行调查分析、结果显示土壤螨类无论是个体数量还是相对丰度,都具有明显的表聚性特点,85％的土壤螨类皆出现于枯枝落叶层和0～5cm的土层。土壤螨类的季节变动调查显示,在枯枝落叶层,土壤螨类有显著的季节变动,且高峰值出现在1993年10月,在10～15cm的土层中,土壤螨类也有相对显著的季节变动,但它的高峰值出现在1994年1月,而在0～5cm和5～10cm的2个土层中,土壤螨类的数量在各个月份之间没有显著的差异。     

华北地区土壤水分的时空变化特征

土壤水分是重要的水文参数,也是水循环、气候变化等研究的基本要素。本研究利用中国气象局新一代自动土壤水分观测网逐小时土壤水分观测数据,分析2013—2019年间华北地区土壤水分的时空分布和变化趋势及其与降水和温度的关系。结果表明: 研究期间,华北地区10~100 cm土层土壤水分整体呈波动下降趋势,尤以100 cm根区土壤水分减少最明显。华北地区不同深度土壤水分空间分布均呈东南高、西北低的特征。10 cm土层63%面积缺墒。不同深度土壤水分随季节变化明显,夏季各层土壤水分达到最高,墒情适宜,春季土壤水分处于低值。土壤水分与降水和温度均具有较好的响应关系,随着土壤深度的增加相关性逐渐减弱,且土壤水分对降水的响应比对气温的响应更显著。     

基于BP神经网络与ETM+遥感数据的盐城滨海自然湿地覆被分类

高效而精确的湿地遥感分类是大范围湿地资源动态监测与管理的必要保障。本研究使用ETM 遥感数据,借助Matlab神经网络工具箱,构建了基于BP神经网络的滨海湿地覆被分类模型,并将其应用于江苏盐城沿海湿地珍禽国家级自然保护区的核心区的自然湿地覆被分类研究中。本研究选择3、4、7、8波段作为输入层变量,单隐藏层设为10个节点,输出层变量对应待划分的8种覆被类型,构建三层式BP神经网络滨海湿地覆被分类模型。结果显示,BP分类总精度为85.91%,Kappa系数为0.8328,与最小距离法和极大似然法的分类总精度相比,分别提高了7.99%和6.08%,Kappa系数也相比提高。研究结果表明,BP神经网络分类法是一种较为有效的湿地遥感影像分类技术,能够提高分类精度。     

Self-organization of spiking neural network that generates autonomous behavior in a real mobile robot

In this paper, we propose self-organization algorithm of spiking neural network (SNN) applicable to autonomous robot for generation of adoptive and goal-directed behavior. First, we formulated a SNN model whose inputs and outputs were analog and the hidden unites are interconnected each other. Next, we implemented it into a miniature mobile robot Khepera. In order to see whether or not a solution(s) for the given task(s) exists with the SNN, the robot was evolved with the genetic algorithm in the environment. The robot acquired the obstacle avoidance and navigation task successfully, exhibiting the presence of the solution. After that, a self-organization algorithm based on a use-dependent synaptic potentiation and depotentiation at synapses of input layer to hidden layer and of hidden layer to output layer was formulated and implemented into the robot. In the environment, the robot incrementally organized the network and the given tasks were successfully performed. The time needed to acquire the desired adoptive and goal-directed behavior using the proposed self-organization method was much less than that with the genetic evolution, approximately one fifth.     

Quantifying Protein-Protein Interactions of Peripheral Membrane Proteins by Fluorescence Brightness Analysis

Fluorescently labeled proteins that are found both in the cytoplasm and at the plasma membrane, such as peripheral membrane proteins, create stratified fluorescent layers that present a challenging environment for brightness studies with fluorescence fluctuation spectroscopy. The geometry of each layer along with fluorescence and brightness contributions from adjacent layers generates a convoluted raw brightness that conceals the underlying brightness of each individual layer. Because the brightness at a layer establishes the oligomeric state of the fluorescently labeled protein at said layer, we developed a method that connects the experimental raw brightness with the physical brightness at each layered compartment. The technique determines the oligomerization in each compartment from an axial intensity scan through the sample, followed by a fluorescence fluctuation spectroscopy measurement at each layer. We experimentally verify the technique with H-Ras-EGFP as a model system and determine its oligomeric state at both the plasma membrane and in the cytoplasm. Furthermore, we study the oligomerization of the Gag matrix domain of Human T-lymphotropic virus Type 1. The matrix domain targets the Gag polyprotein to the plasma membrane where, subsequently, viral assembly occurs. We determine the oligomerization of matrix in the cytoplasm and observe the onset of protein-protein interactions at the membrane. These observations shed light on the early assembly steps of the retrovirus.     

城市公园植被特征对陆生鸟类集团的影响

城市公共绿地是城市生态系统中重要的鸟类栖息地,其植被特征对鸟类集团存在显著影响。在通过分析植被特征对陆生鸟类集团的作用,从而为公园合理配置植被来提高其作为野生动物栖息地的生态服务功能提供理论基础。2009年10月至2011年10月,采用样线法对上海滨江森林公园进行鸟类调查,利用主成分分析划分鸟类集团,用高度定义植被层次,用卡方检验分析鸟类行为在植被层次上的差异。结果表明,滨江森林公园陆生鸟类群落在乔木层的栖息行为和运动行为频次显著多于其在灌木层和地被层的行为频次,在地被层的取食行为频次显著多于其在乔木层和灌木层的行为频次。陆生鸟类可划分为8个鸟类集团,鸟类集团之间存在栖息、运动和取食空间生态位的重叠。食虫拾取集团、杂食拾取集团、食肉飞取集团和植食拾取集团在栖息、运动和取食空间生态位上均存在较高的重叠度,其通过食性分离各自空间生态位。食虫探取集团和食虫飞取集团互为栖息空间生态位重叠度最高集团,其通过取食方式的不同来实现生态位的分离。根据公园植被特征对鸟类集团的影响结果对上海市公园绿地植被配置提出了建议。     

Quantifying Protein-Protein Interactions of Peripheral Membrane Proteins by Fluorescence Brightness Analysis

Fluorescently labeled proteins that are found both in the cytoplasm and at the plasma membrane, such as peripheral membrane proteins, create stratified fluorescent layers that present a challenging environment for brightness studies with fluorescence fluctuation spectroscopy. The geometry of each layer along with fluorescence and brightness contributions from adjacent layers generates a convoluted raw brightness that conceals the underlying brightness of each individual layer. Because the brightness at a layer establishes the oligomeric state of the fluorescently labeled protein at said layer, we developed a method that connects the experimental raw brightness with the physical brightness at each layered compartment. The technique determines the oligomerization in each compartment from an axial intensity scan through the sample, followed by a fluorescence fluctuation spectroscopy measurement at each layer. We experimentally verify the technique with H-Ras-EGFP as a model system and determine its oligomeric state at both the plasma membrane and in the cytoplasm. Furthermore, we study the oligomerization of the Gag matrix domain of Human T-lymphotropic virus Type 1. The matrix domain targets the Gag polyprotein to the plasma membrane where, subsequently, viral assembly occurs. We determine the oligomerization of matrix in the cytoplasm and observe the onset of protein-protein interactions at the membrane. These observations shed light on the early assembly steps of the retrovirus.     

Identification of different isoforms of 14-3-3 protein family in human dermal and epidermal layers

We have previously demonstrated a high level of stratifin, also known as 14-3-3 sigma in differentiated keratinocyte cell lysate and conditioned medium (CM). In this study, we asked the question of whether other 14-3-3 isoforms are expressed in human dermal fibroblasts, keratinocytes, intact dermal and epidermal layers of skin. In order to address this question, total proteins extracted from cultured cells or skin layers were subjected to western blot analysis using seven different primary antibodies specific to well-known mammalian isoforms, beta, gamma, epsilon, eta, sigma, tau, and zeta of 14-3-3 protein family. The autoradiograms corresponding to each isoform were then quantified and compared. The results revealed the presence of very high levels of all seven isoforms in cultured keratinocyte and conditioned medium. With the exception of tau isoform, other 14-3-3 isoforms were also present in intact epidermal layer of normal skin. The profile of 14-3-3 proteins in whole skin was similar to that of epidermis. In contrast, only gamma 14-3-3 isoform, was present in dermal layer obtained from the same skin sample. On the other hand, cultured fibroblasts express a high level of beta, epsilon, gamma and eta and a low level of zeta and tau, but not sigma isoform. However, the levels of 14-3-3 epsilon, gamma and eta were barely detectable in fibroblast conditioned medium. Further, we also used immunohistochemical staining to identify the 14-3-3 isoform expressing cells in human skin sections. The finding revealed different expression profile for each of these isoforms mainly in differentiated keratinocytes located within the layer of lucidum. However, fibroblasts located within the dermal layer did not show any detectable levels of these proteins. In conclusion, all members of 14-3-3 proteins are expressed by cells of epidermal but not dermal layer of skins and that these proteins are mainly expressed by differentiated keratinocytes.     

A combined RSA and FE study of the implanted proximal tibia: correlation of the post-operative mechanical environment with implant migration

There is strong evidence to suggest that inducible displacements, migration and implant loosening are closely related to the initial mechanical environment of the implanted tibia. If this is true, then it should be possible to predict the likelihood of implant migration using patient-specific finite element models. Finite element models of the proximal implanted tibiae were analysed based on pre-operative quantitative computed tomography data of four patients entered into a radiographic migration study. These four patients were also part of an radiostereometric analysis (RSA) study. A variety of load cases were analysed and the risk of bone failure determined for a 2 mm layer of bone immediately beneath the tibial tray. The results were compared with the RSA data measured 1 year post-operatively for each patient. For each patient, an appropriate load case was selected based on patient weight and on the varus-valgus migrations observed in the migration study. The two patients with press-fit implants were predicted to have the highest risk of failure and were found to migrate the most. The two patients with bonded implants (one HA coated and one cemented) were found to have a low risk of failure and these implants migrated the least. This study suggests that the degree of implant migration is dependent on the initial mechanical environment and can be determined using patient-specific finite element analysis.     

The motion of the left ventricle—III

We use the concept of a layered wall, where each separate layer is to be homogeneous, isotropic, and incompressible, to derive stress-strain relations for the middle layer muscle ring at the transverse midsection of the left ventricle; a convenient method of formulation is that based on the elastic potential function. The hoop or circumferential stress in all three layers is found using dimensional and mechanical parameters derived earlier. The various parameters are expressed as Fourier series so that their behavior over a complete ventricular cycle is known analytically. The cases of simple elongation and what we termcurvilinear simple elongation are considered for the middle layer muscle ring strain, and the resulting stress-strain relations are derived. The results are compared with an incompressible rubber-like material known as a Mooney material.