Excitation energy transfer in the light-harvesting chlorophyll a/b.protein

The "light-harvesting chlorophyll a/b.protein" described by Thornber has been prepared electrophoretically from spinach chloroplasts. The optical properties relevant to energy transfer have been measured in the red region (i.e. 600-700 nm). Measurements of the absorption spectrum, fluorescence excitation spectrum and excitation dependence of the fluorescence emission spectrum of this protein confirm that energy transfer from chlorophyll b to chlorophyll a is highly efficient, as is the case in concentrated chlorophyll solutions and in vivo. The excitiation dependence of the fluorescence polarization shows a minimum polarization of 1.9% at 650 nm which is the absorption maximum of chlorophyll b in the protein and rises steadily to a maximum value of 13.8% at 695 nm, the red edge of the chlorophyll a absorption band. Analysis of these measurements shows that at least two unresolved components must be responsible for the chlorophyll a absorption maximum. Comparison of polarization measurements with those observed in vivo shows that most of the depolarization observed in vivo can take place within a single protein. Circular dichroism measurements show a double structure in the chlorophyll b absorption band which suggest an exciton splitting not resolved in absorption. Analysis of these data yields information about the relative orientation of the So leads to S1 transition moments of the chlorophyll molecules within the protein.     

The photochemical and fluorescence properties of whole cells, spheroplasts and spheroplast particles from the blue-green alga Phormidium luridum.

The photochemical activities and fluorescence properties of cells, spheroplasts and spheroplast particles from the blue-green alga Phormidium luridum were compared. The photochemical activities were measured in a whole range of wavelengths and expressed as quantum yield spectra (quantum yield vs. wavelength). The following reactions were measured. Photosynthesis (O2 evolution) in whole cells; Hill reaction (O2 evolution) with Fe(CN)63- and NADP as electron acceptors (Photosystem II and photosystem II + Photosystem I reactions); electron transfer from reduced 2,6-dichlorophenolindophenol to diquat (Photosystem I reaction). The fluorescence properties were emission spectra, quantum yield spectra and the induction pattern. On the basis of comparison between the quantum yield spectra and the pigments compositions the relative contribution of each pigment to each photosystem was estimated. In normal cells and spheroplasts it was found that Photosystem I (Photosystem II) contains about 90% (10%) of the chlorophyll a, 90% (10%) of the carotenoids and 15% (85%) of the phycocyanin. In spheroplast particles there is a reorganization of the pigments; they loose a certain fraction (about half) of the phycocyanin but the remaining phycocyanin attaches itself exclusively to Photosystem I (!). This is reflected by the loss of Photosystem II activity, a flat quantum yield vs. wavelength dependence and a loss of the fluorescence induction. The fluorescence quantum yield spectra conform qualitatively to the above conclusion. More quantitative estimation shows that only a fraction (20--40%) of the chlorophyll of Photosystem II is fluorescent. Total emission spectrum and the ratio of variable to constant fluorescence are in agreement with this conclusion. The fluorescence emission spectrum shows characteristic differences between the constant and variable components. The variable fluorescence comes exclusively from chlorophyll a; the constant fluorescence is contributed, in addition to chlorophyll a, by phycocyanine and an unidentified long wavelength component. The variable fluorescence does not change in the transition from whole cells to spheroplasts. However, the constant fluorescence increases considerably. This indicates the release of a small fraction of pigments from the photosynthetic photochemical apparatus which then become fluorescent.     

基于叶绿素荧光参数的小麦叶片叶绿素相对含量估算方法

叶绿素含量是植物学和农业相关研究领域常用的生理指标.叶绿素含量和叶片光合功能密切相关,但是现有的叶绿素含量的测定方法无法实现叶绿素含量和光合功能的同步测定和关联分析.为解决该问题,本研究通过测定35个小麦品种旗叶的SPAD值和叶绿素荧光诱导动力学曲线,分别使用不同时间的快速叶绿素荧光动力学曲线的荧光值,以及33个常用荧...     

Ratios as a size adjustment in morphometrics

Simple ratios in which a measurement variable is divided by a size variable are commonly used but known to be inadequate for eliminating size correlations from morphometric data. Deficiencies in the simple ratio can be alleviated by incorporating regression coefficients describing the bivariate relationship between the measurement and size variables. Recommendations have included: 1) subtracting the regression intercept to force the bivariate relationship through the origin (intercept-adjusted ratios); 2) exponentiating either the measurement or the size variable using an allometry coefficient to achieve linearity (allometrically adjusted ratios); or 3) both subtracting the intercept and exponentiating (fully adjusted ratios). These three strategies for deriving size-adjusted ratios imply different data models for describing the bivariate relationship between the measurement and size variables (i.e., the linear, simple allometric, and full allometric models, respectively). Algebraic rearrangement of the equation associated with each data model leads to a correctly formulated adjusted ratio whose expected value is constant (i.e., size correlation is eliminated). Alternatively, simple algebra can be used to derive an expected value function for assessing whether any proposed ratio formula is effective in eliminating size correlations. Some published ratio adjustments were incorrectly formulated as indicated by expected values that remain a function of size after ratio transformation. Regression coefficients incorporated into adjusted ratios must be estimated using least-squares regression of the measurement variable on the size variable. Use of parameters estimated by any other regression technique (e.g., major axis or reduced major axis) results in residual correlations between size and the adjusted measurement variable. Correctly formulated adjusted ratios, whose parameters are estimated by least-squares methods, do control for size correlations. The size-adjusted results are similar to those based on analysis of least-squares residuals from the regression of the measurement on the size variable. However, adjusted ratios introduce size-related changes in distributional characteristics (variances) that differentially alter relationships among animals in different size classes. © 1993 Wiley-Liss, Inc.     

基于挺水植物高光谱信息的再生水总氮含量估测——以北京市门城湖湿地公园为例

高光谱信息是探测植物体内氮素含量状况的重要手段,而植物体中的氮素与水体含氮量息息相关.本研究区为以再生水为主要补给水水源的北京门城湖湿地公园,通过获取区内典型的再生水氮净化挺水植物芦苇和香蒲叶片的高光谱数据,并在室内测定对应样点的水体总氮含量指标, 探讨基于典型湿地挺水植物高光谱数据对水体总氮进行遥感探测的可行性.采用4种高光谱参数(光谱指数、归一化差值指数、“三边”参数及吸收特征参数)分别建立一元线性模型、逐步多元回归模型和偏最小二乘模型,根据决定系数(R²)和均方根误差(RMSE)进行模型精度检验.结果表明: 逐步多元回归和偏最小二乘模型的预测精度高于一元线性模型. 3种模型对芦苇的拟合效果均优于香蒲.偏最小二乘模型对芦苇的拟合效果最优(R²=0.854,RMSE=0.647).500～700 nm是反映水氮含量的最佳波段范围,绿峰与红谷反射率的比值与水体总氮含量具有较强的相关性,尤其是吸收特征参数能够较好地预测水体总氮含量.     

The photochemical and fluorescence properties of whole cells,spheroplasts and spheroplast particles from the blue-green alga Phormidium luridum

The photochemical activities and fluorescence properties of cells, spheroplasts and spheroplast particles from the blue-green alga Phormidium luridum were compared. The photochemical activities were measured in a whole range of wavelengths and expressed as quantum yield spectra (quantum yield vs. wavelength). The following reactions were measured: Photosynthesis (O₂ evolution) in whole cells; Hill reaction (O₂ evolution) with Fe(CN)₆^3? and NADP as electron acceptors (Photosystem II and Photosystem II+Photosystem I reactions); electron transfer from reduced 2,6-dichlorophenolindophenol to diquat (Photosystem I reaction). The fluorescence properties were emission spectra, quantum yield spectra and the induction pattern.On the basis of comparison between the quantum yield spectra and the pigments compositions the relative contribution of each pigment to each photosystem was estimated. In normal cells and spheroplasts it was found that Photosystem I (Photosystem II) contains about 90 % (10 %) of the chlorophyll a, 90 % (10 %) of the carotenoids and 15 % (85 %) of the phycocyanin. In spheroplast particles there is a reorganization of the pigments: they loose a certain fraction (about half) of the phycocyanin but the remaining phycocyanin attaches itself exclusively to Photosystem I (!). This is reflected by the loss of Photosystem II activity, a flat quantum yield vs. wavelength dependence and a loss of the fluorescence induction.The fluorescence quantum yield spectra conform qualitatively to the above conclusion. More quantitative estimation shows that only a fraction (20–40 %) of the chlorophyll of Photosystem II is fluorescent. Total emission spectrum and the ratio of variable to constant fluorescence are in agreement with this conclusion.The fluorescence emission spectrum shows characteristic differences between the constant and variable components. The variable fluorescence comes exclusively from chlorophyll a; the constant fluorescence is contributed, in addition to chlorophyll a, by phycocyanine and an unidentified long wavelength component.The variable fluorescence does not change in the transition from whole cells to spheroplasts. However, the constant fluorescence increases considerably. This indicates the release of a small fraction of pigments from the photosynthetic photochemical apparatus which then become fluorescent.     

Simulation of PSII-operating efficiency from chlorophyll fluorescence in response to light and temperature in chrysanthemum (<Emphasis Type="Italic">Dendranthema grandiflora</Emphasis>) using a multilayer leaf model

Chlorophyll fluorescence serves as a proxy photosynthesis measure under different climatic conditions. The objective of the study was to predict PSII quantum yield using greenhouse microclimate data to monitor plant conditions under various climates. Multilayer leaf model was applied to model fluorescence emission from actinic light-adapted (F') leaves, maximum fluorescence from light-adapted (Fm') leaves, PSII-operating efficiency (F_q'/F_m'), and electron transport rate (ETR). A linear function was used to approximate F' from several measurements under constant and variable light conditions. Model performance was evaluated by comparing the differences between the root mean square error (RMSE) and mean square error (MSE) of observed and predicted values. The model exhibited predictive success for Fq'/Fm' and ETR under different temperature and light conditions with lower RMSE and MSE. However, prediction of F' and Fm' was poor due to a weak relationship under constant (R² = 0.48) and variable (R² = 0.35) light.     

Excitation energy transfer in the light-harvesting chlorophyll

The “light-harvesting chlorophyll a/b · protein” described by Thornber has been prepared electrophoretically from spinach chloroplasts. The optical properties relevant to energy transfer have been measured in the red region (i.e. 600–700 nm). Measurements of the absorption spectrum, fluorescence excitation spectrum and excitation dependence of the fluorescence emission spectrum of this protein confirm that energy transfer from chlorophyll b to chlorophyll a is highly efficient, as is the case in concentrated chlorophyll solutions and in vivo. The excitation dependence of the fluorescence polarization shows a minimum polarization of 1.9 % at 650 nm which is the absorption maximum of chlorophyll b in the protein and rises steadily to a maximum value of 13.8 % at 695 nm, the red edge of the chlorophyll a absorption band. Analysis of these measurements shows that at least two unresolved components must be responsible for the chlorophyll a absorption maximum. Comparison of polarization measurements with those observed in vivo shows that most of the depolarization observed in vivo can take place within a single protein. Circular dichroism measurements show a doublet structure in the chlorophyll b absorption band which suggests an exciton splitting not resolved in absorption. Analysis of these data yields information about the relative orientation of the S₀→S₁ transition moments of the chlorophyll molecules within the protein.     

Simultaneous determination of gemcitabine and its metabolite in human plasma by high-performance liquid chromatography

Gemcitabine (dFdC) is a pyrimidine antimetabolite with broad spectrum activity against tumors. In this paper, a normal-phase high-performance liquid chromatographic method was developed for the determination of the parent drug (dFdC) and its metabolite (dFdU) in human plasma. The described sample preparation procedure for determination of dFdC and dFdU is rapid, sensitive, reproducible and simple. The linear regression equations obtained by least square regression method, were area under the curve=0.0371 concentration (ng ml(-1))+192.53 and 1.05.10(-4) concentration (ng ml(-1))-1.2693 for dFdC and dFdU, respectively. The assay for dFdC and dFdU described in the present report has been applied to plasma samples from a bladder cancer patient.     

Use of a Microscope Photometer To Analyze In Vivo Fluorescence Intensity of Epilithic Microalgae Grown on Artificial Substrata

An epifluorescence microscope photometer was used to develop a new, in vivo fluorimetric method for analyzing fluorescence intensities of epilithic microalgae grown on clay tiles in the field. This enabled a nondestructive, direct quantification of algal biomass on the substratum surface. Measurements of a chlorophyll a standard in ethanol (90%) with our fluorimetric method (exitation at 546 nm; emission, >590 nm) correlated well with those from conventional spectrofluorimetric and spectrophotometric methods. Biofilms were analyzed with the microscope photometer by measuring the in vivo fluorescence intensity of 70 spots distributed randomly over the tile surface. They were then analyzed by the two in vitro methods after photopigment extraction. Chlorophyll a content and in vivo fluorescence intensity correlated well. The regression curves were linear up to 6 (mu)g cm(sup-2) but were quadratic or hyperbolic at higher concentrations of up to 28 (mu)g cm(sup-2). The degree of scatter among individual measurements was higher in biofilms than chlorophyll a standards. This in vivo analysis is well suited to ecological experiments and has the advantage of measuring on an extremely small scale, which enables direct analysis of the microdistribution of epilithic microalgae in live biofilms. We demonstrated this by comparing fluorescence intensities of the grazing tracks of the snail Ancylus fluviatilis with those of ungrazed areas. Our in vivo analysis is also unique in enabling biofilms on artificial substrata to be removed, analyzed, and then returned intact in field or laboratory experiments.     

Sensor combination and chemometric variable selection for online monitoring of Streptomyces coelicolor fed-batch cultivations

Fed-batch cultivations of Streptomyces coelicolor, producing the antibiotic actinorhodin, were monitored online by multiwavelength fluorescence spectroscopy and off-gas analysis. Partial least squares (PLS), locally weighted regression, and multilinear PLS (N-PLS) models were built for prediction of biomass and substrate (casamino acids) concentrations, respectively. The effect of combination of fluorescence and gas analyzer data as well as of different variable selection methods was investigated. Improved prediction models were obtained by combination of data from the two sensors and by variable selection using a genetic algorithm, interval PLS, and the principal variables method, respectively. A stepwise variable elimination method was applied to the three-way fluorescence data, resulting in simpler and more accurate N-PLS models. The prediction models were validated using leave-one-batch-out cross-validation, and the best models had root mean square error of cross-validation values of 1.02 g l⁻¹ biomass and 0.8 g l⁻¹ total amino acids, respectively. The fluorescence data were also explored by parallel factor analysis. The analysis revealed four spectral profiles present in the fluorescence data, three of which were identified as pyridoxine, NAD(P)H, and flavin nucleotides, respectively.     

A retrieval algorithm to evaluate the Photosystem I and Photosystem II spectral contributions to leaf chlorophyll fluorescence at physiological temperatures

A new computational procedure to resolve the contribution of Photosystem I (PSI) and Photosystem II (PSII) to the leaf chlorophyll fluorescence emission spectra at room temperature has been developed. It is based on the Principal Component Analysis (PCA) of the leaf fluorescence emission spectra measured during the OI photochemical phase of fluorescence induction kinetics. During this phase, we can assume that only two spectral components are present, one of which is constant (PSI) and the other variable in intensity (PSII). Application of the PCA method to the measured fluorescence emission spectra of Ficus benjamina L. evidences that the temporal variation in the spectra can be ascribed to a single spectral component (the first principal component extracted by PCA), which can be considered to be a good approximation of the PSII fluorescence emission spectrum. The PSI fluorescence emission spectrum was deduced by difference between measured spectra and the first principal component. A single-band spectrum for the PSI fluorescence emission, peaked at about 735?nm, and a 2-band spectrum with maxima at 685 and 740?nm for the PSII were obtained. A linear combination of only these two spectral shapes produced a good fit for any measured emission spectrum of the leaf under investigation and can be used to obtain the fluorescence emission contributions of photosystems under different conditions. With the use of our approach, the dynamics of energy distribution between the two photosystems, such as state transition, can be monitored in vivo, directly at physiological temperatures. Separation of the PSI and PSII emission components can improve the understanding of the fluorescence signal changes induced by environmental factors or stress conditions on plants.     

Fluorescence emission spectra of photosystem I, photosystem II and the light-harvesting chlorophyll a/b complex of higher plants

Fluorescence emission spectra excited at 514 and 633 nm were measured at -196 degrees C on dark-grown bean leaves which had been partially greened by a repetitive series of brief xenon flashes. Excitation at 514 nm resulted in a greater relative enrichment of the 730 nm emission band of Photosystem I than was obtained with 633 nm excitation. The difference spectrum between the 514 nm excited fluorescence and the 633 nm excited fluorescence was taken to be representative of a pure Photosystem I emission spectrum at -196 degrees C. It was estimated from an extrapolation of low temperature emission spectra taken from a series of flashed leaves of different chlorophyll content that the emission from Photosystem II at 730 nm was 12% of the peak emission at 694 nm. Using this estimate, the pure Photosystem I emission spectrum was subtracted from the measured emission spectrum of a flashed leaf to give an emission spectrum representative of pure Photosystem II fluorescence at -196 degrees C. Emission spectra were also measured on flashed leaves which had been illuminated for several hours in continuous light. Appreciable amounts of the light-harvesting chlorophyll a/b protein, which has a low temperature fluorescence emission maximum at 682 nm, accumulate during greening in continuous light. The emission spectra of Photosystem I and Photosystem II were subtracted from the measured emission spectrum of such a leaf to obtain the emission spectrum of the light-harvesting chlorophyll a/b protein at -196 degrees C.     

Estimation of muscle active state

A method is presented for the estimation of the complete time course of muscle active state. The method is based on the selection of a proper model for the muscle, consisting of linear and non-linear components, and on the estimation of its parameters from a simple experiment. The model's parameters are estimated, using the least square method, from measurements of a tetanized muscle's response to a change of its length. The time course of the active state is calculated from an isometric twitch tension response of the same muscle. The twitch tension response is taken as the system's output, and the active state as its input. The latter can be estimated since the system parameters have already been estimated from the tetanized muscle experiment. Experiments were performed on the gastrocnemius muscle of frogs and cats. Results are given for the whole active state time course of these muscles. The results show that the peak active state force does not reach tetanic value, and a negative force is generated during the relaxation period. Additional experiments were carried out with the purpose of verifying the existence of this force; however, no conclusive results were obtained.This research was supported by the Julius Silver Institute of Bio-Medical Engineering Sciences, Grant 050-304     

Comparing Analysis Methods in Functional Calcium Imaging of the Insect Brain

We investigate four different methods for background estimation in calcium imaging of the insect brain and evaluate their performance on six data sets consisting of data recorded from two sites in two species of moths. The calcium fluorescence decay curve outside the potential response is estimated using either a low-pass filter or constant, linear or polynomial regression, and is subsequently used to calculate the magnitude, latency and duration of the response. The magnitude and variance of the responses that are obtained by the different methods are compared, and, by computing the receiver operating characteristics of a classifier based on response magnitude, we evaluate the ability of each method to detect the stimulus type and conclude that a polynomial approximation of the background gives the overall best result.     

Estimation and comparison of changes in the presence of informative right censoring: conditional linear model

A general linear regression model for the usual least squares estimated rate of change (slope) on censoring time is described as an approximation to account for informative right censoring in estimating and comparing changes of a continuous variable in two groups. Two noniterative estimators for the group slope means, the linear minimum variance unbiased (LMVUB) estimator and the linear minimum mean squared error (LMMSE) estimator, are proposed under this conditional model. In realistic situations, we illustrate that the LMVUB and LMMSE estimators, derived under a simple linear regression model, are quite competitive compared to the pseudo maximum likelihood estimator (PMLE) derived by modeling the censoring probabilities. Generalizations to polynomial response curves and general linear models are also described.     

Estimation of the density of nymphs of the bush tick, Haemaphysalis longicornis (Acari: Ixodidae), by the catch effort method

The density of nymphs of the bush tick, Haemaphysalis longicornis, was investigated by the catch effort method with flagging. The spatial distribution of H. longicornis nymphs fit the model of contagiously distributed colonies by Iwao's m*-m analysis (Iwao 1968). A sequential sampling method was used to predict the theoretical point at which to stop sampling. Our analysis showed that five quadrats (4 m x 4 m) were sufficient to estimate the density of H. longicornis nymphs with a mean density of 5.39 per quadrat. We estimated the tick density by two methods with respect to the sampling interval. The estimated density of ticks based on ticks collected during short sampling intervals (within a half hour) was 511.34 in the 18 quadrats with the extrapolation of the linear regression equation. On the other hand, for the long interval sampling, the total number of ticks estimated by the linear regression equation was 635.47 in six quadrats in which ticks had been collected by long interval sampling. There was a significant difference between the slopes of the two linear regression equations, suggesting that the rate of reduction in the number of H. longicornis nymphs in the study area by the catch effort method differed between the two sampling methods.     

Fluorescence emission spectra of Photosystem I,Photosystem II and the light-harvesting chlorophyll a/b complex of higher plants

Fluorescence emission spectra excited at 514 and 633 nm were measured at ?196 °C on dark-grown bean leaves which had been partially greened by a repetitive series of brief xenon flashes. Excitation at 514 nm resulted in a greater relative enrichment of the 730 nm emission band of Photosystem I than was obtained with 633 nm excitation. The difference spectrum between the 514 nm excited fluorescence and the 633 nm excited fluorescence was taken to be representative of a pure Photosystem I emission spectrum at ?196 °C. It was estimated from an extrapolation of low temperature emission spectra taken from a series of flashed leaves of different chlorophyll content that the emission from Photosystem II at 730 nm was 12% of the peak emission at 694 nm. Using this estimate, the pure Photosystem I emission spectrum was subtracted from the measured emission spectrum of a flashed leaf to give an emission spectrum representative of pure Photosystem II fluorescence at ?196 °C. Emission spectra were also measured on flashed leaves which had been illuminated for several hours in continuous light. Appreciable amounts of the light-harvesting chlorophyll a/b protein, which has a low temperature fluorescence emission maximum at 682 nm, accumulate during greening in continuous light. The emission spectra of Photosystem I and Photosystem II were subtracted from the measured emission spectrum of such a leaf to obtain the emission spectrum of the light-harvesting chlorophyll a/b protein at ?196 °C.     

以小时为步长的大兴安岭典型林分地表死可燃物含水率模型预测及外推精度

地表死可燃物含水率是火险天气和火行为预报中的重要指标.本研究基于时滞平衡含水率法(Nelson和Simard方法)及气象要素回归方法,于2010年9—10月对黑龙江省大兴安岭地区盘古林场不同郁闭度的山杨-白桦混交林、红皮云杉纯林,以及采伐迹地(原1∶1樟子松-白桦混交林)地表死可燃物含水率进行以小时为步长的连续测定,建立其预测模型,得到预测误差,并使用相应的模型对其他林分地表死可燃物含水率进行外推精度分析.结果表明:采用Nelson平衡含水率法构建的地表死可燃物含水率变化模型的平均绝对误差、平均相对误差和均方误差根(0.0154、0.104和0.0226)低于Simard法(0.0185、0.117和0.0256)和气象要素回归法(0.0222、0.150和0.0331).在外推效果方面,气象要素回归法的平均绝对误差、平均相对误差和均方误差根(0.0410、0.0300和0.0740)低于Simard法(0.610、0.492和0.846),但前两者均高于Nelson法(0.034、0.021和0.0660),说明以小时为步长的时滞平衡含水率法,尤其是Nelson法适用于大兴安岭地区所测林分.外推虽不能降低误差,但有助于提高现有模型应用至不同林分条件或大尺度范围内的地表死可燃物含水率预测精度和利用率.模型建模和外推误差与不同树种和郁闭度条件差异有关,研究时应根据不同林分和地点选择合适的平衡含水率模型.