Formação de biofilmes por Enterococcus faecium sensível e multirresistente aos antimicrobianos: características à proteômica

Enterococcus faecium tem se destacado no cenário das infecções hospitalares, particularmente, amostras portadoras de características de multirresistência. Apesar de serem responsabilizados como agentes etiológicos em diferentes quadros clínicos, fatores associados a patogênese das infecções ainda não estão esclarecidos. Entretanto, sabe-se que a capacidade de formação de biofilmes pode ser responsabilizada como um dos atributos capazes de promover o papel patogênico desses microrganismos. Assim sendo, este estudo se propôs a investigar a capacidade de formação de biofilmes por duas amostras de E. faecium: SS-1274 (derivada da amostra tipo da espécie) e CL-6729 (multirresistente e pertencente a um complexo clonal globalmente disperso). As amostras foram caracterizadas fenotipica e genotipicamente para confirmação da identificação, determinação da susceptibilidade a 17 antimicrobianos, caracterização da concentração mínima inibitória para ampicilina, gentamicina e vancomicina, determinação do genótipo vanA e detecção de genes associados a expressão dos genes asa1, cylA, esp, gelE e hyl. A análise quantitativa da formação por 24h e 72h dos biofilmes foi realizada por metodologia do cristal violeta, em placas de microtitulação de poliestireno. Foram construídas curvas de formação pela avaliação da DO570nm das preparações coradas por cristal violeta em períodos de tempo de 2h a 74h. A influência de subCMIs de ampicilina, gentamicina e vancomicina na formação de biofilmes de E. faecium foi também caracterizada em ensaios de quantificação da biomassa. A presença de DNA e proteínas foi avaliada em ensaios de destacamento e por fluorimetria. A arquitetura, distribuição espacial e reação aos fluorocromos Syto9 e SYPRO Ruby Protein foram evidenciadas por microscopia confocal de varredura a laser. Análises proteômicas através da avaliação por SDS-PAGE e por ESI-Q-TOF também foram empregadas para avaliação de biofilmes versus crescimento planctônico. Nossos resultados demonstraram que a amostra CL-6729 apresentou uma maior biomassa nos tempos analisados. Apesar da menor quantidade de biomassa da amostra SS-1274, o perfil da curva de formação foi semelhante a CL-6729. As análises da matriz por ensaios quantitativos e microscopia confocal revelaram que proteína parece ser um importante constituinte para ambas as amostras, entretanto biofilmes formados na presença de da CMI e durante 24h, parecem sofrer alterações na constituição. Os resultados relativos às espectrometrias de massa sugerem que células de biofilmes de E. faecium podem também estar metabolicamente ativas, devido a identificação de um considerável número de proteínas relacionadas ao metabolismo e divisão celular, similarmente às células planctônicas. No entanto, investigações complementares são necessárias para quantificar as diferenças relacionadas a sua expressão. Foi observado que ambas as amostras independente das variações fenotípicas e genotípicas são capazes de formar biofilmes maduros exibindo constituição e arquitetura similares. Entretanto, nossos resultados sugeriram que cada amostra responde as diferentes situações de acordo com seus determinantes de virulência e resistência.

Avaliação das alterações macro-hemodinâmicas, microcirculatórias, gasométricas, metabólicas e inflamatórias secundárias à sedação com dexmedetomidina em um modelo experimental de endotoxemia em hamsters

Pela sua alta incidência, morbidade, mortalidade e custos ao sistema de saúde, a sepse se destaca entre as diversas indicações de internação em unidade de terapia intensiva (UTI). A disfunção da microcirculação tem papel central na gênese e manutenção da síndrome séptica, sendo um marco fisiopatológico desta síndrome. Pacientes críticos invariavelmente estão ansiosos, agitados, confusos, desconfortáveis e/ou com dor. Neste contexto, drogas sedativas são amplamente utilizadas na medicina intensiva. A dexmedetomidina, um agonista potente e altamente seletivo dos receptores alfa-2 adrenérgicos, vem conquistando espaço como o sedativo de escolha nas UTIs por seus efeitos de sedação consciente, redução da duração e incidência de delirium e preservação da ventilação espontânea. Apesar destas possíveis vantagens, a indicação de uso da dexmedetomidina na síndrome séptica ainda carece de conhecimentos sobre seus efeitos na microcirculação e perfusão orgânica. Com o intuito de caracterizar os efeitos microcirculatórios da dexmedetomidina em um modelo murino de endotoxemia que permite estudos in vivo da inflamação e disfunção da perfusão microvascular, hamsters Sírios dourados submetidos à endotoxemia induzida por administração intravenosa de lipopolissacarídeo de Escherichia coli (LPS, 1,0 mg.kg-1) foram sedados com dexmedetomidina (5,0 μg.kg.h-1). A microscopia intravital da preparação experimental (câmara dorsal) permitiu a realização de uma análise quantitativa das variáveis microvasculares e do rolamento e adesão de leucócitos à parede venular. Também foram analisados os parâmetros macro-hemodinâmicos e gasométricos (arterial e venoso portal), as concentrações de lactato arterial e venoso portal, a água pulmonar total e a sobrevivência do animal. Animais não-endotoxêmicos e/ou tratados com solução salina a 0,9% serviram como controles neste experimento. O LPS aumentou o rolamento e a adesão de leucócitos à parede venular, diminuiu a densidade capilar funcional e a velocidade das hemácias nos capilares e induziu acidose metabólica. O tratamento com dexmedetomidina atenuou significativamente estas respostas patológicas (p < 0,05). A frequência de pulso dos animais foi significativamente reduzida pela droga (p < 0,05). Outros resultados não foram tão expressivos (estatisticamente ou clinicamente). Estes resultados indicam que a utilização de dexmedetomidina produz um efeito protetor sobre a microcirculação da câmara dorsal de hamsters endotoxêmicos. Efeitos anti-inflamatórios da dexmedetomidina sobre os leucócitos e o endotélio poderiam melhorar a perfusão capilar e representar o mecanismo in vivo de ação da droga na microcirculação.

判别分析系统应运于生物标本鉴定的初步研究

计算机技术和数学方法、手段应用于生物标本鉴定的研究工作在国外开展已有多年，但在国内却并没有许多人涉猎，也没有受到足够的重视。 鉴于这一领域的重要性及其实际意义，本文综合、扩充、改进了多元统计判别分析和模糊模式识别中的多种定量化判别方法，初步在计算机上实现了一个可用于生物标本鉴定或其它与判别、识别有关方面的系统，并将其用于桔梗科沙参属三个种：泡沙参、多歧沙参和裂叶沙参及菖蒲科中两个种的标本鉴定上面，获得了比较满意的判别效果。同时，为了弥补数量化标本鉴定的不足，本文作者还设计开发了一个描述性的基于检索表的人机交互式的标本鉴定模块。另外，本系统还包括模糊系统聚类和典型相关分析等模块，可供生物分类及其它定量分析运算时选用。 对于以上各种判别、识别方法的差异及优劣，文中根据实例做出了综合分析和比较，并认为在所有方法当中，逐步判别法和模糊协方差识别法最适宜于生物标本鉴定之用。最后，作者展望了未来计算机用于生物标本鉴定的前景。

鄂尔多斯高原景观要素空间格局及植物与环境关系的定量解析

鄂尔多斯高原是一个多层次、复杂的生态过渡带，具有复杂多样的环境条件、生态特点，因此也就具有复杂多样的植物与环境关系。本文从群落和景观两个尺度水平上研究鄂尔多斯高原植物或植被与环境关系及景观空间格局。利用鄂尔多斯高原野外植物群落样方调坦数据、微生境环境数据、气候数据，以典范对应分析（Canonical Correspondence Analysis, CCA）的方法分析了鄂尔多斯高原植物分布空间格局与环境要素的关系并对秋类环境要素对鄂尔多斯高原植物空间分布格局的贡献进行了定量分解;利用1:500 000 鄂尔多斯高原植被、土壤、土地利用、土地沙漠化类型等专题地图在GIS支持下分析了鄂尔多斯高原景观空间格局、并利用上述专题图数据加上鄂尔多斯高原气候数据库分析了土壤、土地利用、土地利用、土地沙漠化和气候等对鄂尔多斯高原植被空间分布格局的作用。通过分析，得到了以下主要结论： 1 在分析方法上、利用典范对应分析的方法，把植物分布的空间因素与环境因素分离的方法发展为植物分布空间格局不同类型影响因素作用的定量分离，提出了相应的概念模型和实现方法。 2 影响鄂尔多斯高原植物分布空间格局的主要微生境环境要素是基质类型、地下水位、覆沙厚度等，而影响鄂尔多斯高原分布空间格局的主要气候要素中，降水和干湿指标的作用大于温度和热量指标的作用。 3 通过对鄂尔多斯高原植物分布空间格局与环境关系的研究，以植物对微生境环境要素的反应为根据，把鄂尔多斯高原主要植物划分为4个大类群：梁地植物、沙地植物、草甸植物和耐盐植物。根据它们对气候要素的反应，把鄂尔多斯高原主要植物划分为典型草原植物、荒漠草原植物和草原化荒漠植物3大类。进一步，根据鄂尔多劳动保护高原植物与环境关系的研究，进行了鄂尔多斯高原植被功能型划分的尝试，得到了鄂尔多斯高原的12种主要植被功能型。 4 对鄂尔多斯高原植物分布的空间格局的影响环境因素的贡献作了定量地分解。分析结果显示：鄂尔多斯高原植物分布空间格局中有27.02％可由已知环境变量得到解释，其中21.56％与微生境环境要素相关，7.51％与气候要素的作用有关，而气候与微生境环境要素的耦合作用的份额为2.05％。根据植物生长是否直接受到地下水的影响，鄂尔多斯高原存在两大类型生态特点差异明显的生境类型：中性立地和隐域生境，对两大类型生境上影响植物空间分布格局的环境要素的作用也进行了定量分解。分析结果表明：对于植物生长不直接受地下水影响的中性立地，已知环境要素的作用可以解释植物空间分布格局总信息的29.36％，稍大于对总体上鄂尔多斯高原植物分布空间格局的解释，其中9.23％与气候要素相关，22.08％与微生境环境要素相关，而两种类型环境要素的耦合作用则占1.95％。对于植物生长直接受到地下水影响的隐域生境，所有已知环境要素对植物分布空间格局的贡献率为72.28％，其中气候要素的作用为30.31％，微生境环境要素的作用为49.08％，两类环境要素的耦合作用为7.11％。 5 描述景观空间格局的指数多种多样，这些能数在描述特定区域的景观空间格局时是有信息冗余的。本文对利用FRAGSTATS所获得的鄂尔多斯高原植被、土壤、土地利用、土地沙漠化等景观分量的20个景观指数实施了因子分析。通过因子分析，我们可以把描述鄂尔多斯高原景观空间格局的景观指数归并为以下8类：多样性指数、斑块多度指数、斑块类型丰富度指数、斑块面积指数、斑块形状指数、分形维数、空间配置指数和斑块面积变异指数。通过因子分析，还得到了这些景观指数对描述鄂尔多斯景观格局的共性特征：在描述鄂尔多斯高原景观空间格局时，作用最大的是多样性指数、斑块多度指数、面积加权平均斑块形状指数和面积加权平均分形维数，其次是斑块类型丰富度指数、平均分维指数、平均形状指数和斑块面积指数，而空间配置指数（扩散与毗连指数）和斑块面积变异指数的作用则比较微弱。 6 对鄂尔多斯高原景观指数的因子分析是非常有效和成功的。因子分析对鄂尔多斯高原植被、土壤、土地利用、土地沙漠化景观指数的分析分别得到了5-6个主要因子，可以表达原有20个景观指数所表达信息的91.1-96.0％，即可以反应鄂尔多斯高原景观空间格局的大部分信息。本文所进行的因子分析对因子进行了方差最大化(Varimax)正交旋转的处理，因子分析得到的每一个主要因子都有一个或几个与之相关性非常高的景观指数与之对应，因此，就可以用与因子分析所得主要因子相关性最高的景观指数代替该主要因子来表达鄂尔多斯高原的景观空间格局。另外还因为有些景观指数之间具有极高的相关系数，所以对因子分析所得到的景观指数可以进一步精减，最后利用因子分析成功地把原有20个景观指数减少到了11个。最后被选来描述鄂尔多斯高原景观格局的景观指数有下列11个：MSIEI（修正的Simpson均匀度指数）、AWMPFD（面积加权平均斑块分形维数）、AWMSI（面积加权平均形状指数）、NP（斑块数目）、PR（斑块类型丰富度）、MSI（平均形状指数）、MPFD（平均斑块分形维数）、MPS（平均斑块面积）、PSCV（斑块面积变异系数）、DLFD（双对数分形维数）和IJI（扩散与毗连指数）。 7 鄂尔多斯高原植被、土壤、土地利用在景观组成结构上具有一个共同特点，就是各种类型的面积差异极大，少数类型占有极大比重，而其余面积则很小。产生这一情形的原因主要与人为活动的强烈影响有关，表现在地带性的植被与土壤面积所占的比重不高，沙地、沙生植被与风沙土则占有很大比重。 8 以地带性植被和滩地隐域性植被表示的鄂尔多斯高原的原生植被仅占高原面积的不足30％，而以地带性土壤和滩地隐域性土壤表示的原生性土壤占鄂尔多斯高原总面积的近40％，说明土壤退化不如植被退化严重，或滞后于植被退化。 9 鄂尔多斯高原各景观指数的空间变化曲线，植被与土壤很相近，具有非常相似的格局;土地利用景观格局空间变化特征与植被、土壤等明显不同;土地沙漠化的景观格局空间变化曲线介植被曲线、土壤曲线与土地利用曲线之间，说明土地沙漠化不仅是一个受人为活动影响的过程，而且与自然过程密切相关。 10 鄂尔多斯高原景观格局的空间梯度变化表现出了东西向和南北向的梯度，但总体上以东西向的变化比较明显。 11 通过鄂尔多斯高原土壤类型、土地利用、土地沙漠化等景观要素、气候、空间要素与鄂尔多斯高原植被空间分布格局的CCA分析，探讨了它们之间的相互关系。以对鄂尔多斯高原植被组成数据的总方差解释的百分率为标准，土壤对鄂尔多斯植被分布的空间格局的作用最大，其方差贡献率可达44.28％，其次是土地利用与鄂尔多斯高原植被的关系也很密切，土地利用对鄂尔多斯高原植被空间分布格局的方差贡献率为22.45％，空间因素对鄂尔多斯高原植被空间分布格局的贡献率为17.51％，土地沙漠化对鄂尔多斯高原植被空间分布格局的贡献为15.65％，排在第四位;气候因素对鄂尔多斯高原植被空间格局的贡献率为11.95％，居第五位。 12 在气候要素对鄂尔多斯高原植被空间分布格局的作用中，降水与干湿指标的作用大于温度与热量指标的作用。这一点与利用野外调查样方的群落数据植物与气候关系的分析是完全一致的。CCA分析还表明鄂尔多斯高原植被空间格局的东西向变化大于南北向分异。 13 在群落和景观水平上，鄂尔多斯高原植物空间分布或植被格局的影响因素的作用具有相似的格局，即气候因子的作用明显地小于地质、土壤、水文等微生境环境要素（群落水平）或土壤（景观水平）的作用，并在这两个尺度上气候要素对植物空间分布或植被格局的定量解释份额上也是非常相近的，都仅有10％左右。气候因子对鄂尔多斯高原植物空间分布格局的这种弱的解释能力，从侧面说明了人为活动等非自然因素对鄂尔多斯高原植物空间分布格局的强烈作用。 14 在鄂尔多斯高原生态系统管理上，应协调人与自然的关系;加强鄂尔多斯高原的生物多样性保育，对于本区生态和经济对非常重要的滩地，应协调好对其开发利用与保护的关系;在鄂尔多斯高原土地沙漠化防治方面，应把调整人地关系与自然生态背景与条件相结合，如使用“三圈”模式等生态系统管理模式等。

植物种群小尺度空间结构的数量分析

采用位置指数、分异指数、结构复杂性指数、Shannon-Wiener多样性指数以及Ripley的K-方程，探讨了华南海岸英罗港红树植物木榄种群的分布格局、胸围和树高分异以及冠层结构方面的空间异质性。结果表明，多数木榄种群呈现随机分布，其个体胸围和树高的分异程度较低;而少数木榄种群呈现集群分布，其个体胸围和树高的分异程度明显。采用地理信息系统对木榄种群冠层和空隙斑块进行多种水平和垂直尺度的分析，冠层与空隙斑块之间的镶嵌格局因种群而异，这种格局可基于树冠投影用Shannon-Wiener多样性指数进行定量描述。冠层结构的空问异质性随空间尺度而变化，但这种变化在一定尺度范围内保持相对的稳定。这一尺度范围可作为木榄红树林更新或生态管理单位的参考尺度。 采用多重分形理论和方法对华南海岸北海红树林区红树植物白骨壤幼苗种群的分布格局进行了分析。结果表明，白骨壤幼苗种群的分布格局具有多标量和多重分形行为。随着q值由-2增加到4，Dq值介于1.078-1.997，f(a)值介于0.402-1.678，a(q)值介于0.909-2.480。多重分形的参数值与幼苗种群的集聚强度密切相关。差值（D。-D1）和[f(-1) –D]。是度量格局空间异质性程度的有效指数。海洋水文周期性规律、不同面积大小的裸地、树冠的阻拦作用等是制约白骨壤幼苗种群多重分形格局形成的主要因子。 采用地统计学理论和方法，对北京东灵山落叶松和胡桃楸针阔混交林下的克隆植物绢毛匍匐委陵菜分株种群的统计变量和土壤变量的空间格局进行了分析和比较。结果表明，绢毛匍匐委陵菜分株种群各器官生物量以及分株数和叶数的空间格局具有较高程度的异质性。总生物量以及茎、叶和根生物量的半方差函数曲线为指数模型，叶柄生物量、分株数和叶数的为球状模型，叶片生物量的为线性无基台值模型，而匍匐茎生物量的为纯块金效应模型。除叶片和匍匐茎的生物量外，其它器官生物量以及分株数和叶数由空间自相关引起的空间异质性．SHA主要表现在120-3 50cm的尺度范围内，SHA占总空间异质性的比例在50%以上。生境中，各种土壤变量的空间格局也具有较高程度的异质性。土壤含水量、NO3-、NH4+、有机质、全磷和PO4(3-)的半方差函数曲线为球状模型，而全氮、K+和pH的为指数模型。土壤变量由空间自相关引起的空间异质性SHA主要表现在8-lOOcm的尺度范围内，SHA占总空间异质性的比例在68%以上。绢毛匍匐委陵菜分株种群空间格局的自相关尺度显著大于其生境中土壤变量空间格局的自相关尺度，表明绢毛匍匐委陵菜通过匍匐茎相互联结的克隆分株种群对异质性土壤资源表现出较大的缓冲能力。绢毛匍匐委陵菜分株数的空间格局与土壤的含水量和全氮空间格局之间在几乎所有尺度上的相关性都不显著，但与其它土壤变量空间格局之间都存在不同程度的相关性，其中与NH4+、pH、PO4(3-)和全磷空间格局之间的相关性尺度范围较为明显和较大。通过分株数、叶数和生物量分配的可塑性变化，绢毛匍匐委陵菜分株种群的空间格局与土壤资源的空间格局之间产生相互联系，并随空间尺度的变化会发生改变。

Studies on fish lipids. Pt. 2. Fatty acid composition of lipids of oil sardine (Sardinella longiceps)

Gas-liquid chromatography has been employed for the qualitative and quantitative analysis of the component fatty acids in lipids of oil sardine (Sardinella longiceps). Phospholipids and triglycerides of the lipids were previously separated by column chromatography before they were converted into the methyl esters of the fatty acids. The predominant acids present in the depot fat of the fish have been found to be C14:0=8.13%, C16:0=27.9%, C18:0=3.8%, C18:1=15.4%., C20:5=10.6% and C22:6=8.8%. Apart from the above acids the distribution of minor acids belonging to Cl8, C20 and C22 groups have also been worked out. The separated phospholipid fraction contained more than 70% polyunsaturated acids of which the important constituents were docosahexaenoic (C22:6=28%) and eicosapentaenoic (C20:5=10.6%). A marked reduction was found in the amounts of polyunsaturated acids in triglycerides, their total amount registering about 20%. This fraction recorded about 48% of C16 acids of which palmitic and palmitoleic acids amounted to 25.8% and 19.1% respectively. Occurrence of odd numbered fatty acids C15 and C17 has also been noted in the phospholipid and composite samples of the fish.

How to achieve ultra high photoconductive gain for transparent oxide semiconductor image sensors

We present quantitative analysis of the ultra-high photoconductivity in amorphous oxide semiconductor (AOS) thin film transistors (TFTs), taking into account the sub-gap optical absorption in oxygen deficiency defects. We analyze the basis of photoconductivity in AOSs, explained in terms of the extended electron lifetime due to retarded recombination as a result of hole localization. Also, photoconductive gain in AOS photo-TFTs can be maximized by reducing the transit time associated with short channel lengths, making device scaling favourable for high sensitivity operation. © 2012 IEEE.

Liquid fuel visualization using laser-induced fluoresence during cold start

The presence of liquid fuel inside the engine cylinder is believed to be a strong contributor to the high levels of hydrocarbon emissions from spark ignition (SI) engines during the warm-up period. Quantifying and determining the fate of the liquid fuel that enters the cylinder is the first step in understanding the process of emissions formation. This work uses planar laser induced fluorescence (PLIF) to visualize the liquid fuel present in the cylinder. The fluorescing compounds in indolene, and mixtures of iso-octane with dopants of different boiling points (acetone and 3-pentanone) were used to trace the behavior of different volatility components. Images were taken of three different planes through the engine intersecting the intake valve region. A closed valve fuel injection strategy was used, as this is the strategy most commonly used in practice. Background subtraction and masking were both performed to reduce the effect of any spurious fluorescence. The images were analyzed on both a time and crank angle (CA) basis, showing the time of maximum liquid fuel present in the cylinder and the effect of engine events on the inflow of liquid fuel. The results show details of the liquid fuel distribution as it enters the engine as a function of crankangle degree, volatility and location in the cylinder. A. semi-quantitative analysis based on the integration of the image intensities provides additional information on the temporal distribution of the liquid fuel flow. © 1998 Society of Automotive Engineers, Inc.

Joint experimental and numerical approach to three-dimensional shock control bump research

Previous studies of transonic shock control bumps have often been either numerical or experimental. Comparisons between the two have been hampered by the limitations of either approach. The present work aims to bridge the gap between computational fluid dynamics and experiment by planning a joint approach from the outset. This enables high-quality validation data to be produced and ensures that the conclusions of either aspect of the study are directly relevant to the application. Experiments conducted with bumps mounted on the floor of a blowdown tunnel were modified to include an additional postshock adverse pressure gradient through the use of a diffuser as well as introducing boundary-layer suction ahead of the test section to enable the in-flow boundary layer to be manipulated. This has the advantage of being an inexpensive and highly repeatable method. Computations were performed on a standard airfoil model, with the flight conditions as free parameters. The experimental and computational setups were then tuned to produce baseline conditions that agree well, enabling confidence that the experimental conclusions are relevant. The methods are then applied to two different shock control bumps: a smoothly contoured bump, representative of previous studies, and a novel extended geometry featuring a continuously widening tail, which spans the wind-tunnel width at the rear of the bump. Comparison between the computational and experimental results for the contour bump showed good agreement both with respect to the flow structures and quantitative analysis of the boundary-layer parameters. It was seen that combining the experimental and numerical data could provide valuable insight into the flow physics, which would not generally be possible for a one-sided approach. The experiments and computational fluid dynamics were also seen to agree well for the extended bump geometry, providing evidence that, even though thebumpinteracts directly with the wind-tunnel walls, it was still possible to observe the key flow physics. The joint approach is thus suitable even for wider bump geometries. Copyright © 2013 by S. P. Colliss, H. Babinsky, K. Nubler, and T. Lutz. Published by the American Institute of Aeronautics and Astronautics, Inc.

Subcellular localization and characterization of G protein-coupled receptor homolog from lymphocystis disease virus isolated in China

G protein-coupled receptors (GPCRs) constitute a large superfamily involved in various types of signal transduction pathways, and play an important role in coordinating the activation and migration of leukocytes to sites of infection and inflammation. Viral GPCRs, on the other hand, can help the virus to escape from host immune surveillance and contribute to viral pathogenesis. Lymphocystis disease virus isolated in China (LCDV-C) contains a putative homolog of cellular GPCRs, LCDV-C GPCR. In this paper, LCDV-C GPCR was cloned, and the subcellular localization and characterization of GPCR protein were investigated in fish cells. LCDV-C GPCR encoded a 325-amino acid peptide, containing a typical seven-transmembrane domain characteristic of the chemokine receptors and a conserved DRY motif that is usually essential for receptor activation. Transient transfection of GPCR-EGFP in fathead minnow (FHM) cells and epithelioma papulosum cyprini (EPC) cells indicated that LCDV-C GPCR was expressed abundantly in both the cytoplasm and nucleoplasm. Transient overexpression of GPCR in these two cells cannot induce obvious apoptosis. FHM cells stably expressing GPCR showed enhanced cell proliferation and significant anchorage-independent growth. The effects of GPCR protein on external apoptotic stimuli were examined. Few apoptotic bodies were observed in cells expressing GPCR treated with actinomycin D (ActD). Quantitative analysis of apoptotic cells indicated that a considerable decrease in the apoptotic fraction of cells expressing GPCR, compared with. the control cells, was detected after exposure to ActD and cycloheximide. These data suggest that LCDV-C GPCR may inhibit apoptosis as part of its potential mechanism in mediating cellular transformation.

Electron microscopic examination of the viromatrix of Rana grylio virus in a fish cell line

Rana grylio virus (RGV), a Ranavirus belonging to the family Iridoviridae, assembles in the viromatrix which is a factory for viral genome replication and particle assembly. Ultrastructural studies of the viromatrix will clarify the pathway of assembly. The viromatrix and quantitative changes in RGV infected epithelipma papulosum cyprini (EPC) cells, one of fish cell lines, were studied by electron microscopy. It was shown that viromatrices were adjacent to the nucleus, and the electron density was lower than that of the surrounding cytoplasm. The viromatrix contained virus particles with different forms, electron-dense materials and amorphous structures which included tubules and membranous materials. Tubules were often observed in direct continuity with empty capsids. Several bundles of intermediate filaments were seen alongside the viromatrix and crystalline aggregates. Large clusters of mitochondria occurred in proximity to viromatrix. A total of 990 cells profiles were examined. The results showed that 394 cells contained viromatrix: 89.3% contained one, and 10.7% contained two to four viromatrices. The number of viromatrices increased gradually and reached a peak at 16 h p.i. The viromatrix area at 24 h p.i. increased up to 7.4 +/- 0.69 mu m(2) which was three-times lower than that at 6 h p.i. The number of empty capsids within viromatrix was generally more than that of "full" particles at different time points, and there was a strong positive correlation between them. (c) 2005 Elsevier B.V. All rights reserved.

Rapid patterning of 1-D collagenous topography as an ECM protein fibril platform for image cytometry.

Cellular behavior is strongly influenced by the architecture and pattern of its interfacing extracellular matrix (ECM). For an artificial culture system which could eventually benefit the translation of scientific findings into therapeutic development, the system should capture the key characteristics of a physiological microenvironment. At the same time, it should also enable standardized, high throughput data acquisition. Since an ECM is composed of different fibrous proteins, studying cellular interaction with individual fibrils will be of physiological relevance. In this study, we employ near-field electrospinning to create ordered patterns of collagenous fibrils of gelatin, based on an acetic acid and ethyl acetate aqueous co-solvent system. Tunable conformations of micro-fibrils were directly deposited onto soft polymeric substrates in a single step. We observe that global topographical features of straight lines, beads-on-strings, and curls are dictated by solution conductivity; whereas the finer details such as the fiber cross-sectional profile are tuned by solution viscosity. Using these fibril constructs as cellular assays, we study EA.hy926 endothelial cells' response to ROCK inhibition, because of ROCK's key role in the regulation of cell shape. The fibril array was shown to modulate the cellular morphology towards a pre-capillary cord-like phenotype, which was otherwise not observed on a flat 2-D substrate. Further facilitated by quantitative analysis of morphological parameters, the fibril platform also provides better dissection in the cells' response to a H1152 ROCK inhibitor. In conclusion, the near-field electrospun fibril constructs provide a more physiologically-relevant platform compared to a featureless 2-D surface, and simultaneously permit statistical single-cell image cytometry using conventional microscopy systems. The patterning approach described here is also expected to form the basics for depositing other protein fibrils, seen among potential applications as culture platforms for drug screening.

1994-2006年《水土保持研究》载文被引定量分析

利用中国知识资源总库的中国引文数据库检索《水土保持研究》在1994-2006年所载论文的被引用情况,采用文献计量学方法对该刊1994-2006年载文被《中国期刊文献引证数据库》来源期刊引用的情况进行统计分析及评价。从文献引证的角度透视《水土保持研究》的学术水平。该刊1994-2006年共载文2098篇,被引用2018篇,占载文总数的96.62%。2001-2006年该刊影响因子、即年指标等文献计量指标逐年上升,分析表明:《水土保持研究》载文质量较高,在该学科有较大的学术影响力。

基于GIS的黄土丘陵区土壤水库库容组成及其定量分析——以陕北安塞县为例

以空间图形和数据库为基础,对土壤水库的相关技术指标、研究深度和静态库容组成等进行了描述、界定和计算。研究认为:安塞县5m深土层土壤水库总库容为1419.78mm/416156万m3,其中死库容占土壤总库容的21.08%,重力库容占土壤总库容的13.82%,有效库容占土壤总库容的65.10%,最大有效库容占总库容的78.92%;从土地利用类型方面来看,坡耕地和荒坡地总库容量最大,分别占研究区土壤水库总库容的37.65%和36.04%;从坡度分级方面来看,>25°和10°～15°坡度级别土壤总库容量最大,分别占研究区土壤水库总库容的41.50%和30.52%;峁坡和沟坡土壤水库库容组成基本相等。

县级尺度土地利用结构特征定量分析

基于土地利用数量和空间结构内涵理解,选取相关定量指标组合并结合GIS技术,以安徽省含山县为例,利用其2002年土地变更数据和2000年1︰50000土地利用现状图,探讨县级尺度土地利用结构特征定量分析方法。研究表明:定量分析结果与县域实际相一致,所选取的定量指标能够充分发挥各自作用,尤其是在表征数量结构总体特征上,多样化指数、集中化指数、洛伦兹曲线和组合系数4个指标之间优势互补,相互验证,最为显著,这说明定量分析具有可行性,其关键在于合理地选取具有明确意义的定量指标组合,包括指标选取以及各指标之间的优化组合。全县土地利用结构总体特征可概括为:以耕地-林地-未利用地-水域为组合类型,以耕地和林地为景观基质,土地利用程度不高,以农用地为主,县域地形地貌格局是土地利用结构形成的自然基础,单位面积土地GDP、人口密度与土地利用程度、建设用地区位优势在一定程度上呈正相关。该研究可为县级,尤其是类似地区土地利用结构特征定量分析提供借鉴和参考。