Fiber-Optic Fluorometer for Microscale Mapping of Photosynthetic Pigments in Microbial Communities

Microscale fluorescence measurements were performed in photosynthetic biofilms at a spatial resolution of 100 to 200 μm with a new fiber-optic fluorometer which allowed four different excitation and emission wavelengths and was configured for measuring phycobiliproteins, chlorophylls, and bacteriochlorophylls. We present details of the measuring system and describe examples of applications in different microbial communities.     

How Should Thiopurine Treatment be Monitored?— Methodological Aspects

Monitoring of thiopurine metabolites is important due to a complex metabolism with large interindividual variation, but the suitability of currently used methods has been questioned. The drawbacks include poor reproducibility, the inability to differentiate between the different analytes, as well as the use of a nontarget matrix. Further research should be directed toward measuring thiopurine metabolites in mononuclear cells, measuring the different nucleotides specifically, as well as measuring the incorporation of thioguanine into DNA. The studies should not be limited to thioguanosine nucleotides but include methylthioinosine nucleotides as well.     

The influence of variations in the measuring procedure on quantitative nuclear image features in histologic sections of lung tissue

The qualitative and quantitative features of cell nuclei in tissue sections play an important role in diagnostic histopathology; variations in staining intensity and measuring procedures may interfere with their proper evaluation. To identify nuclear features that are relatively insensitive to these technical variables, the influence of critical steps in a scanning-stage densitometer measuring system was studied on 87 quantitative nuclear image (QNI) features in histologic sections of lung tissue. The influences of the following measuring variations were evaluated: interactive segmentation (with and without median filtering; with and without 5% uniform distributed noise added); scanning (with and without median filtering); calibration of the photomultiplier (different background localizations and different intensity levels); and time. In addition, the influence of artificially changed intensity variations was investigated. The results showed that, while the coefficient of variation (CV) induced by variations in the measuring system was usually low (below 10%), for some QNI features the CV can be high (up to 216%). The influence of artificial variations in intensity was restricted: only a minority of the QNI features showed a significant difference. Of the 87 QNI features, 35 had a CV of less than 10%, and 25 of these were significantly correlated with each other. Thus, only ten uncorrelated, low-CV QNI features remained; these belonged to all of the different QNI feature categories used. These features may be diagnostically important since they may best describe the morphologic properties of the nuclei. The results of this study should help in selecting quantitative nuclear image features that are less sensitive to variations in the measuring procedure and staining intensity.     

用于胃癌检测的漫反射光谱

胃癌正严重地威胁人类的健康,为了开发一种可以诊断早期胃癌的新型光学检测技术,建立了一套简便的组织体漫反射光谱检测装置.首先,根据组织体的光学特性,介绍光谱检测方法的基本原理.然后,利用组织癌变导致的组织体光学特性变化,建立漫反射光谱检测的实验装置.最后,利用该实验装置分别获取正常胃组织和胃癌组织的漫反射光谱.实验结果表明:癌变和正常胃组织的漫反射光谱在可见光区域有明显差别,特别是在波长为500 nm和630 nm处,但是漫反射光谱检测无法分辨来自组织不同层的光谱信息.这些结果说明漫反射光谱检测装置可用于胃癌的辅助检测.     

pH track of expanding bacterial populations

A method of pH distribution measurements in agar nutrient media containing expanding bacterial populations is described. It is based on measuring pH microsamples taken at different points of the media. The sample volume was 10 microliters. A pH sensitive field effect transistor was used as a measuring electrode. Acidification was found to occur in glucose media, while alkalization occurred in the media containing peptone.     

Measuring electrical impedance of organs--instrumental equipment for research and clinical use

An apparatus for measuring the impedance of intact biological organs or parts of organs in the frequency range of 10 Hz to 10 MHz is described. In this range impedance exhibits a large dispersion, which is dependent on tissue structures. The time course of alterations of electrical impedance such as occur during ischemia can be recorded with this equipment. Five specimens in five measuring chambers can be examined simultaneously at different temperatures. In the second part of the article, a portable impedance meter for measuring the modulus of impedance near 200 Hz, the phase of impedance at 5 kHz and the local temperature at the measuring point, is described. These parameters permit an intra-operative evaluation of the changing state of ischemic organs. Sterilizable probes with four surface electrodes and an integrated temperature sensor permit atraumatic measurements at the organ surface. The measurement itself is harmless to the tissue.     

Is the microwave irradiation a suitable method for measuring soil microbial biomass?

Soil microbial biomass (SMB), the living part of soil organic matter, is used to quantify the total biomass of microorganisms present in the soil. The importance of studies about SMB has emphasized the need to identify methods which can measure the size of SMB. Among the methods currently available, chloroform-fumigation extraction and incubation are the most commonly used for estimation of SMB. However, several studies have proposed the microwave (MW) irradiation as a quick, simple and safe alternate method. There are different opinions about suitability of this method for measuring SMB. There is a question to do “Is the microwave irradiation a suitable method for measuring soil microbial biomass?” Most of the published papers comparing MW and chloroform-fumigation showed strong relationship between both methods. Therefore, we consider MW a suitable method for measuring SMB; however, it is necessary to calibrate the MW methods in different soils with a range of properties, such as clay content, to find an appropriate conversion factor in order to generate correct values for SMB with MW method.     

Sucrose acceptance threshold: a way to measure sugar perception in ants

Variation in the perception of sweet taste is a well-known phenomenon in the animal kingdom. Well-established protocols for measuring sucrose responsiveness in non-social insects and honeybees have made it possible to understand many aspects of their biology and behaviour. Ants are also advanced social insects that present a plethora of life histories with diverse strategies and behaviours; however, a universal paradigm possible to measure this response in different ant species has not yet been developed. Here, we present a protocol for measuring the sucrose acceptance threshold (SAT) under controlled conditions in harnessed ants with different feeding habits. By testing the response to antennal and palp sucrose stimulation and using the occurrence of licking as the response, we developed a non-ambiguous evaluation that allowed easy detection of threshold changes. The results showed that the response to both antennal and palp stimulation varied widely among species. Some species licked in response to antennal stimulation while others did so in response to palp stimulation. Using the appropriate kind of stimulation, we tested the SAT protocol in ants of different genera and ants of the same species with different levels of sugar reserve. The differences detected in both cases imply that the protocol is appropriate for measuring and detecting variations in sugar perception in ants.     

Comparison of the dye method with the thermocouple psychrometer for measuring leaf water potentials

The dye method for measuring water potential was examined and compared with the thermocouple psychrometer method in order to evaluate its usefulness for measuring leaf water potentials of forest trees and common laboratory plants. Psychrometer measurements are assumed to represent the true leaf water potentials. Because of the contamination of test solutions by cell sap and leaf surface residues, dye method values of most species varied about 1 to 5 bars from psychrometer values over the leaf water potential range of 0 to −30 bars. The dye method is useful for measuring changes and relative values in leaf potential. Because of species differences in the relationships of dye method values to true leaf water potentials, dye method values should be interpreted with caution when comparing different species or the same species growing in widely different environments. Despite its limitations the dye method has a usefulness to many workers because it is simple, requires no elaborate equipment, and can be used in both the laboratory and field.     

Dream Recall Frequency, Attitude Towards Dreams and Openness to Experience

The question whether personality dimensions explain the interindividual differences in dream recall frequency has often been investigated by dream researchers. The present findings confirm previous research which has shown that traits such as openness-to-experience and thin boundaries correlate substantially with dream recall frequency. However, correlation coefficients are small and are much larger if attitude towards dreams or a scale measuring different aspects of dream recall are considered. Thus, future studies should consider the differentiation between items measuring dream recall and related aspects and items measuring attitudes towards dreams. Schonbar's life-style hypothesis should be revised slightly: not dream recall frequency but attitude towards dreams and the way to deal with dreams are part of a broader life style.     

用积分球系统测量大鼠组织在不同光阑孔径下对氦-氖激光的温反射特性

本文对大鼠组织在不同光阑孔径下对He-Ne激光的漫反射特性进行了测量。实验采用了不同光阑孔径的积分球系统及He-Ne激光,并根据测量数据计算和分析了大鼠组织对该波长激光的漫反射特性。结果表明,有些大鼠组织在632.8nm波长的He-Ne激光照射下对不同的光阑孔径其漫反射率没有显著性区别,而有些大鼠组织在632.8nm波长的He-Ne激光照射下对不同的光阑孔径其漫长反射率有显著性区别,提示在测定不同生物组织的光学特性时,应考虑组织样品的受光照射的面积的大小对测定结果的精确度的影响。     

Measurement of relative antioxidant activity of compounds: a methodological note

The relative activities of some hydrogen-donating antioxidants were assessed by comparing their activities with that of Trolox (Trolox equivalent antioxidant capacity, TEAC) for scavenging the ABTS radical cation (ABTS.+) generated in the aqueous phase. We have verified, however, that TEAC values may change with the concentration of compounds and with the measuring times used. Not withstanding, TEAC values do not differ significantly if the compounds have kinetic curves of ABTS.+ formation similar to that of Trolox. This is the case with ascorbic acid, whose TEAC values, determined by using five concentrations at three different measuring times, are very close. For the flavonoids studied (catechin, rutin, naringenin and silibinin) which have kinetic curves of ABTS.+ formation different from that of Trolox, the TEAC values decrease with increasing concentrations of the compounds for each measuring time, and increase with increasing measuring times for each concentration. In the present study, we conclude that, in order to evaluate relative antioxidant activities of compounds by the ABTS assay, it is essential to perform kinetic studies to assess scavenging of ABTS.+ by these compounds. Therefore, when the TEAC values of compounds are determined for more than one measuring time, we may be sure that all the antioxidant potential of compounds is being considered and whether or not it is possible to establish a hierarchy for their antioxidant activities.     

植物细胞凋亡的检测方法

细胞凋亡是一种在动、植物中普遍存在的现象,在动、植物的发育过程中起着非常重要的作用。从形态学、生化及分子生物学、免疫学、生理学等方面介绍了几种检测细胞凋亡的方法,以及各种方法在检测不同植物凋亡时的应用,并对植物细胞凋亡检测技术的前景进行了展望。     

Non-contact measuring of small translations and rotations in all degrees of displacement]

The investigation of biomechanical problems often makes it necessary to measure displacements and rotations in space. An easy-to-handle optical measuring system has been developed that permits non-contact simultaneous determination of small translations and rotations in all degrees of freedom. Three two-dimensional light-sensitive sensors (PSD [position sensing detector] elements) were arranged spatially to form a measuring sensor. After processing in a computer, the signals definitively determine relative shift and rotation. Thus it is possible, for example, to measure the relative movement between two adjacent spinal vertebrae under different loads.     

The light-addressable potentiometric sensor for multi-ion sensing and imaging

The light-addressable potentiometric sensor (LAPS) is a semiconductor-based chemical sensor with an electrolyte-insulator-semiconductor structure. The LAPS can have many measuring points integrated on the sensing surface, which are individually accessed by a light beam. By modifying the measuring points with different materials, a single sensor plate can be used as a multi-analyte sensor. In this paper, instrumentation and application of LAPS to multi-ion sensing and imaging are described. As a new application of LAPS, potentiometric imaging of a microfluidic channel is proposed.     

Temperature dependence of DNA persistence length

We have determined the temperature dependence of DNA persistence length, a, using two different methods. The first approach was based on measuring the j-factors of short DNA fragments at various temperatures. Fitting the measured j-factors by the theoretical equation allowed us to obtain the values of a for temperatures between 5°C and 42°C. The second approach was based on measuring the equilibrium distribution of the linking number between the strands of circular DNA at different temperatures. The major contribution into the distribution variance comes from the fluctuations of DNA writhe in the nicked circular molecules which are specified by the value of a. The computation-based analysis of the measured variances was used to obtain the values of a for temperatures up to 60°C. We found a good agreement between the results obtained by these two methods. Our data show that DNA persistence length strongly depends on temperature and accounting for this dependence is important in quantitative comparison between experimental results obtained at different temperatures.     

Caspase multiplexing: simultaneous homogeneous time-resolved quenching assay (TruPoint) for caspases 1, 3, and 6

Caspases are a group of cysteine proteases involved in apoptosis and inflammation. A multiparametric homogeneous assay capable of measuring activity of three different caspases in a single well of a microtiter plate is described. Different fluorescent europium, samarium, terbium, and dysprosium chelates were coupled to a caspase substrate peptide, their luminescence properties, were analyzed, and their function in a time-resolved fluorescence quenching-based caspase 3 assay was studied. Substrates for caspases 1, 2, 3, 6, and 8 and granzyme B were also synthesized and their specificities for different caspases were determined. By selecting suitable lanthanide chelates and substrates we developed a multiparametric homogeneous time-resolved fluorescence quenching-based assay for caspases 1, 3, and 6. The assay was capable of measuring the activity of both single caspases and a mixture of three caspases mixed in the same well.     

Kinetics of an enzyme-catalyzed reaction measured by electrospray ionization mass spectrometry using a simple rapid mixing attachment

Mass spectrometry offers a potential means of measuring virtually all enzyme-catalyzed reactions by simultaneously measuring the concentrations of substrates, products, and intermediates where there are differences in mass between them. To perform these measurements the reaction mixture must be aged for different times and then ionized. Electrospray ionization mass spectrometry provides the most direct means of measuring these reactions. Here we describe a simple reaction mixing and ageing attachment for an electrospray ionization mass spectrometer, built from commercially available components. We have employed this device to measure the kinetics of a model reaction, namely the hydrolysis of N2-(carbobenzyloxy)-L-lysine-p-nitrophenyl ester-catalyzed by trypsin. In this way we were able to measure the kinetics of substrate depletion, product formation, and changes in both free enzyme and acyl-enzyme intermediate concentration in the approach to steady state. With this device we were able to measure reaction times down to about 640 ms.     

An ancient relation between units of length and volume based on a sphere

The modern metric system defines units of volume based on the cube. We propose that the ancient Egyptian system of measuring capacity employed a similar concept, but used the sphere instead. When considered in ancient Egyptian units, the volume of a sphere, whose circumference is one royal cubit, equals half a hekat. Using the measurements of large sets of ancient containers as a database, the article demonstrates that this formula was characteristic of Egyptian and Egyptian-related pottery vessels but not of the ceramics of Mesopotamia, which had a different system of measuring length and volume units.