A quantitative HPLC method for determining lactide content using hydrolytic kinetics

A new method for quantitative analysis of lactide has been developed by applying chemical kinetics to a HPLC system. The most important advance is its practical approach to the quantification of analytes that are unstable in the HPLC mobile phase. In HPLC analysis, anhydrous mobile phases cannot separate lactide from impurities, and only mixtures of water and organic solvent can achieve effective separation. By selecting conditions for testing and studying the kinetics of lactide hydrolysis, extensive experiments revealed that lactide degradation can be treated as a pseudo-first-order reaction under the given HPLC conditions, and lactide content or purity can be quantitatively determined. This method is practical for measuring the purity of the intermediate lactide in polylactic acid (PLA) production and the lactide content in PLA.

Analysis of norditerpenoid alkalods in processing Radix Aconiti Lateralis preparata with Radix Glycyrrhizae Preparata by electrospray ionization tandem mass spectrometry

Norditerpenoid alkaloids in the processing of Radix aconiti lateralis preparata with Radix glycyrrhizae preparata was studied by electrospray ionization tandem mass spectrometry. A method of ESI-MS couple with internal standard was applied for semi quantitative analysis of norditerpenoid alkaloids before and after processing. The combined-decoction-to single-decoction-ratios of the relative abundance of toxic hypaconitine, mesaconitine and aconitine were 5. 67%, 4. 05% and 4. 88%, respectively. The chemical changes of processing can been comprehensive observed by ESI-MS/MS analysis. The scientific basis of reducing the toxicity of Radix aconiti lateralis preparata with Radix glycyrrhizae preparata was enlightened by compareing with the single-decotion, the combined-decoction and residue for the compatibility of Radix aconiti lateralis preparata and Radix glycyrrhizae preparata.

Analysis of nephroloxic and carcinogenic aristolochic acids in Aristolochia plants by capillary electrophoresis with electrochemical detection at a carbon fiber microdisk electrode

Aristolochic acids (AAs) are the main bioactive ingredients in the most of Aristolochia plants, which are used to make dietary supplements, slimming pills and Traditional Chinese Medicines (TCMs). Excessive ingestion of AAs can lead to serious nephropathy. Therefore, quantitative analysis and quality control for the plants containing AAs is of great importance. In this paper, capillary electrophoresis (CE) with electrochemical detection (ED) at a 33 mu m carbon fiber microdisk electrode (CFE) has been applied to detect AA-I and AA-II in Aristolochia plants. Under the optimum conditions: detection potential at 1.20 V, 2.0 x 10(-2) mol L-1 phosphate buffer solution (PBS) (pH 10.0), injection time 25 s at a height of 17 cm and separation voltage at 12.5 kV, the AA-I and AA-II were baseline separated within 5 min. Low detection limits for AA-I and AA-II were 4.0 x 10(-8) mol L-1 and 1.0 x 10(-7) mol L-1, respectively. Wide linear ranges were from 4.0 x 10(-8) mol L-1 to 1.9 x 10(-5) mol L-1 and 1.0 X 10(-7) mol L-1 to 5.0 x 10(-5) mol L-1 for AA-I and AA-II, respectively. The proposed method has been successfully applied to analyze AAs contents in plant extracts. The results indicated that the contents of AAs in each part of Aristolochia debilis Sieb. Et Zucc.

Study of a new derivatizing reagent that improves the analysis of amino acids by HPLC with fluorescence detection: application to hydrolyzed rape bee pollen

A simple and sensitive method for evaluating the chemical compositions of protein amino acids, including cystine (Cys)(2) and tryptophane (Try) has been developed, based on the use of a sensitive labeling reagent 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl chloroformate (BCEC-Cl) along with fluorescence detection. The chromophore of the 1,2-benzo-3,4-dihydrocarbazole-ethyl chloroformate (BCEOC-Cl) molecule was replaced with the 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl functional group, yielding the sensitive fluorescence molecule BCEC-Cl. The new reagent BCEC-Cl could then be substituted for labeling reagents commonly used in amino acid derivatization. The BCEC-amino acid derivatives exhibited very high detection sensitivities, particularly in the cases of (Cys)(2) and Try, which cannot be determined using traditional labeling reagents such as 9-fluorenyl methylchloroformate (FMOC-Cl) and ortho-phthaldialdehyde (OPA). The fluorescence detection intensities for the BCEC derivatives were compared to those obtained when using FMOC-Cl and BCEOC-Cl as labeling reagents. The ratios I (BCEC)/I (BCEOC) = 1.17-3.57, I (BCEC)/I (FMOC) = 1.13-8.21, and UVBCEC/UVBCEOC = 1.67-4.90 (where I is the fluorescence intensity and UV is the ultraviolet absorbance). Derivative separation was optimized on a Hypersil BDS C-18 column. The detection limits calculated from 1.0 pmol injections, at a signal-to-noise ratio of 3, ranged from 7.2 fmol for Try to 8.4 fmol for (Cys)(2). Excellent linear responses were observed, with coefficients of > 0.9994. When coupled with high-performance liquid chromatography, the method established here allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids including (Cys)(2) and Try from bee-collected pollen (bee pollen) samples.

Column switching-back flushing technique for the analysis of aromatic compounds in gasoline

A simple method, based on the technique of capillary column switching-back flushing, has been developed for the detailed analysis of aromatic compounds in gasoline. The sample was first separated on a 30-m long OV-2330 polar precolumn and then backflushed onto a nonpolar analytical column. The early eluting components from the precolumn and the components of interest (aromatic compounds plus heavier compounds) eluting from the analytical column are all directed to the same flame ionization detection system through a T piece, which permits the quantitative analysis of aromatic hydrocarbons in gasoline by a normalization method using correcting factors. The switching time window of the method is +/-5 s, resulting in easier operation and higher reliability. The reproducibility of the quantitative analysis was less than or equal to3% RSD for real gasoline samples. (C) 2002 Elsevier Science B.V. All rights reserved.

Using oxidized carbon nanotubes as matrix for analysis of small molecules by MALDI-TOF MS

Oxidized carbon nanotubes are tested as a matrix for analysis of small molecules by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Compared with nonoxidized carbon nanotubes, oxidized carbon nanotubes facilitate sample preparation because of their higher solubility in water. The matrix layer of oxidized carbon nanotubes is much more homogeneous and compact than that of nonoxidized carbon nanotubes. The efficiency of desorption/ionization for analytes and the reproducibility of peak intensities within and between sample spots are greatly enhanced on the surface of oxidized carbon nanotubes. The advantage of the oxidized carbon nanotubes in comparison with alpha-cyano-4-hydroxycinnamic acid (CCA) and carbon nanotubes is demonstrated by MALDI-TOF-MS analysis of an amino acid mixture. The matrix is successfully used for analysis of synthetic hydroxypropyl P-cyclodextrin, suggesting a great potential for monitoring reactions and for product quality control. Reliable quantitative analysis of jatrorrhizine and palmatine with a wide linear range (1-100 ng/mL) and good reproducibility of relative peak areas (RSD less than 10 %) is achieved using this matrix. Concentrations of jatrorrhizine (8.65 mg/mL) and palmatine (10.4 mg/mL) in an extract of Coptis chinensis Franch are determined simultaneously using the matrix and a standard addition method. (c) 2005 American Society for Mass Spectrometry.

Quantitative determination of compounds in tobacco essential oils by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry

A quantitative analysis of the individual compounds in tobacco essential oils is performed by comprehensive two-dimensional gas chromatography (GC x GC) combined with flame ionization detector (FID). A time-of-flight mass spectrometer (TOF/MS) was coupled to GC x GC for the identification of the resolved peaks. The response of a flame ionization detector to different compound classes was calibrated using multiple internal standards. In total, 172 compounds were identified with good match and 61 compounds with high probability value were reliably quantified. For comparative purposes, the essential oil sample was also quantified by one-dimensional gas chromatography-mass spectrometry (GC/MS) with multiple internal standards method. The results showed that there was close agreement between the two analysis methods when the peak purity and match quality in one-dimensional GC/MS are high enough. (c) 2005 Elsevier B.V. All rights reserved.

Analytic expression of the diffraction of a circular aperture

Recurring to the characteristic of Bessel function, we give the analytic expression or the Fresnel diffraction by a circular aperture, thus the diffractions on the propagation axis and along the boundary of the geometrical shadow are discussed conveniently. Since it is difficult to embody intuitively the physical meaning from this series expression of the Fresnel diffraction, after weighing the diffractions on the axis and along the boundary of the geometrical shadow, we propose a simple approximate expression of the circular diffraction, which is equivalent to the rigorous solution in the further propagation distance. It is important for the measurement of the parameter or the beam, such as the quantitative analysis of the relationship of the wave error and the divergence of the beam, In this paper, the relationship of the fluctuation of the transverse diffraction profile and the position of the axial point is discussed too. (c) 2005 Elsevier GrnbH. All rights reserved.

Dynamic phase-mapping of domain nucleation on MgO : LiNbO3 crystal by digital holographic interferometry

The quantitative phase-mapping of the domain nucleation in MgO:LiNbO3 crystals is presented by using the digital holographic interferometry. An unexpected peak phase at the beginning of the domain nucleation is observed and it is lowered as the spreading of the domain nucleus. The existence of the nucleus changes the moving speed of the domain wall by pinning it for 3s. Such in-situ quantitative analysis of the domain nucleation process is a key to optimizing domain structure fabrication.

一批中国古玻璃化学成分的质子激发X射线荧光分析

质子激发X射线荧光(proton induced X-ray emission，PIXE)技术是一种高灵敏度、非破坏性、多元素定量测定的分析方法。采用外束PIXE技术对内蒙古地区和博山出土的一批古代玻璃的化学成分进行了定量测定。结果表明：内蒙古地区出土的玻璃中，西周时期的玻璃珠是含有少量K2O和CaO助熔剂的釉砂，其主要成分为SiO2；汉代的玻璃珠属于PbO—SiO2玻璃；多数元代的玻璃制品和部分北魏时期的玻璃珠属于K2O—CaO—SiO2玻璃。博山出土的元末明初的玻璃基本为KzO-CaO—SiO2系玻璃

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。

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.