Image analysis as a tool for quantitative phycology: a computational approach to cyanobacterial taxa identification

In the following work we discuss the application of image processing and pattern recognition to the field of quantitative phycology. We overview the area of image processing and review previously published literature pertaining to the image analysis of phycological images and, in particular, cyanobacterial image processing. We then discuss the main operations used to process images and quantify data contained within them. To demonstrate the utility of image processing to cyanobacteria classification, we present details of an image analysis system for automatically detecting and classifying several cyanobacterial taxa of Lake Biwa, Japan. Specifically, we initially target the genus Microcystis for detection and classification from among several species of Anabaena. We subsequently extend the system to classify a total of six cyanobacteria species. High-resolution microscope images containing a mix of the above species and other nontargeted objects are analyzed, and any detected objects are removed from the image for further analysis. Following image enhancement, we measure object properties and compare them to a previously compiled database of species characteristics. Classification of an object as belonging to a particular class membership (e.g., “Microcystis,”“A. smithii,”“Other,” etc.) is performed using parametric statistical methods. Leave-one-out classification results suggest a system error rate of approximately 3%. Received: September 6, 1999 / Accepted: February 6, 2000     

Reference ranges for urinary concentrations and ratios of endogenous steroids, which can be used as markers for steroid misuse, in a Caucasian population of athletes

The detection of misuse with naturally occuring steroids is a great challenge for doping control laboratories. Intake of natural anabolic steroids alters the steroid profile. Thus, screening for exogenous use of these steroids can be established by monitoring a range of endogenous steroids, which constitute the steroid profile, and evaluate their concentrations and ratios against reference ranges. Elevated values of the steroid profile constitute an atypical finding after which a confirmatory IRMS procedure is needed to unequivocally establish the exogenous origin of a natural steroid. However, the large inter-individual differences in urinary steroid concentrations and the recent availability of a whole range of natural steroids (e.g. dehydroepiandrosterone and androstenedione) which each exert a different effect on the monitored parameters in doping control complicate the interpretation of the current steroid profile.The screening of an extended steroid profile can provide additional parameters to support the atypical findings and can give specific information upon the steroids which have been administered.The natural concentrations of 29 endogenous steroids and 11 ratios in a predominantly Caucasian population of athletes were determined. The upper reference values at 97.5%, 99% and 99.9% levels were assessed for male (n = 2027) and female (n = 1004) populations. Monitoring minor metabolites and evaluation of concentration ratios with respect to their natural abundances could improve the interpretation of the steroid profile in doping analysis.     

Image analysis as a simple and useful tool for evaluating the constitutive heterochromatin (C-banding) in human chromosomes

Image analysis can contribute to those fields of cytogenetics that are influenced most by subjectivity, especially evaluation of chromosome regions that are histochemically polymorphic. C-banding of human or primate chromosomes may be used as a typical example of this concept. In addition, using it quantitatively it is useful in studies of population cytogenetics of man and Primates. Therefore, the aim of this study was to determine the best conditions for measurement by image analysis of C-positive regions in a sample of 29 normal subjects. We looked for relationships of chromosomal C-positive areas with the dimensions of the corresponding metaphases. Finally, we suggest some criteria for potential use of C-banding in population and comparative studies.     

The potential of cloud point system as a novel two-phase partitioning system for biotransformation

Although the extractive biotransformation in two-phase partitioning systems have been studied extensively, such as the water–organic solvent two-phase system, the aqueous two-phase system, the reverse micelle system, and the room temperature ionic liquid, etc., this has not yet resulted in a widespread industrial application. Based on the discussion of the main obstacles, an exploitation of a cloud point system, which has already been applied in a separation field known as a cloud point extraction, as a novel two-phase partitioning system for biotransformation, is reviewed by analysis of some topical examples. At the end of the review, the process control and downstream processing in the application of the novel two-phase partitioning system for biotransformation are also briefly discussed.     

The use of cultured Acer cells as a screening tool for mitotic inhibitors. I. Reference herbicides

Several commercially available mitosis inhibitors have been studied for their effects on an Acer pseudoplatanus cell suspension culture; these were colchicine, N-(1,1-dimethylpropynyl)3-chlorobenzamide, propyzamide and trifluralin. The last two compounds are known as preemergence herbicides. In the studied cultures, the concentrations necessary to arrest mitosis were between 50 and 100 M for colchicine, and between 10 and 100 M for propyzamide, N-(1,1-dimethylpropynyl)3-chlorobenzamide and trifluralin. However, protein synthesis, plastid and mitochondria division and dry weight increase occurred in the treated cells, leading to the formation of giant cells. These results suggest that mitosis inhibition is the fundamental effect of these compounds at the concentrations studied. At these concentrations, metabolic activities seem unaffected, in contrast with what was found previously in the case of chlorpropham. This phenylcarbamate inhibited mitosis in the same cells at 100 M, but also prevented protein synthesis and weight increase, probably as a result of its uncoupling properties.The effect of another compound, glyphosate, on Acer cell suspension cultures, was studied concurrently. At concentrations between 10 and 100 M, it only slightly decreased protein formation in the culture, and was unable to prevent mitosis from occurring.The formation of giant cells in Acer cell suspension cultures seems to be a good criterion to demonstrate that selective mitosis inhibition has taken place. This criterion might be used in the case of mitosis inhibitors acting on the G ₂-phase.

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Résumé Les effets de divers inhibiteurs de la division cellulaire ont été étudiés sur des suspensions cellulaires d' Acer pseudoplatanus. Ces produits sont la colchicine, le N-(1,1-diméthylpropynyl)3-chlorobenzamide, le propyzamide et la trifluraline. Les 2 derniers composés sont des herbicides de pré-levée. Les concentrations nécessaires pour bloquer la mitose dans les suspensions cellulaires se situent de 50 à 100 M pour la colchicine et de 10 à 100 M pour les 3 autres composés.Dans les cellules traitées la synthèse protéique, la multiplication des plastes et des mitochondries et l'augmentation de masse sèche ont lieu, entraînant la formation de cellules géantes. Ces résultats suggèrent que, aux concentrations étudiées, le blocage de la mitose est l'effet primaire de ces produits; les activités métaboliques des cellules, à ces concentrations, ne semble pas affecté contrairement à ce que l'on a observé précédemment avec le chlorprophame. Ce phénylcarbamate, en effet, inhibe la mitose des cellules d'Acer á 100 M mais aussi, et probablement du fait de ses propriétés découplantes, empêche la synthèse protéique et l'augmentation de masse sèche. Le glyphosate étudié parallèlement est incapable de bloquer la mitose entre 10 et 100 M; il freine légèrement la synthèse protéique.L'obtention de cellules géantes dans les suspensions cellulaires d'Acer pseudoplatanus semble un bon test pour mettre en évidence une inhibition sélective de la mitose. Ce critère pourrait être utilisé pour les composés antimitotiques agissant sur la phase G ₂.

     

The use of chromogenic bacteria as coloured substitutes for pathogens: A simple strategy during design and development of a new method for sample pretreatment

Aims: In the present study, chromogenic (red) bacteria were used to simulate actual target bacteria during set‐up and optimization of an isolation process of bacteria, designed for food samples. Isolation of bacteria from food in the context of molecular biological detection of food pathogens is a multistep process. Development of such a separation method requires continuous monitoring of the location of the presumable targets in the sample tubes. Therefore, red‐coloured pigmented bacteria were used as substitutes for the actual target bacteria, during the establishment of a new sample preparation technique. Methods and Results: The chromogenic bacteria Micrococcus roseus and Serratia marcescens were confirmed to withstand the physical (e.g. centrifugal forces) and chemical (e.g. lysis buffer composition) conditions required during establishment of the new technique. Furthermore, the suitability of these model bacteria to substitute for the actual target pathogens (Salmonella enterica subsp. enterica serovar Typhimurium and Listeria monocytogenes) was assured by testing the physical properties of the model bacteria with respect to the proposed separation methods. Conclusion: Visibility of the pigmented bacteria within the complex sample matrices served to allocate bacterial content during the various steps necessary for finalization of the method protocol. The presumptive bacterial targets can be allocated simply by visualization of their bright red colour silhouetted against the background sample matrix. Significance and Impact of Study: The use of pigmented bacteria as substitutes for actual colourless target bacteria during design and development of a bacterial isolation method is a simple and inexpensive application. It saves a huge amount of time and resources, as the proof of principle of new methods is possible in rapid succession.     

The microspore-derived embryo ofBrassica napus L. as a tool for studying embryo-specific lipid biogenesis and regulation of oil quality

Summary A time-course study of lipid accumulation in microspore-derived embryos and developing zygotic embryos of rapeseed (Brassica napus L. ssp.oleifera) is presented. Rapid storage fat (triacylglycerol) biosynthesis was induced in microspore-derived embryos of oilseed rape (cv Topas) when the embryos were transferred from standing cultures (10 ml) to fresh medium (75 ml) and shake cultured. Triacylglycerols accumulated, after a lag period of 7 days, at a linear rate of approximately twice that of the developing zygotic embryo. The fatty acid composition of triacylglycerols in microspore-derived embryos closely parallelled that of the developing zygotic embryos. In the microspore-derived embryos, the amount of phosphatidylcholine, the major substrate for the production of polyunsaturated fatty acids in oilseeds, remained constant during the linear phase of triacylglycerol production, whereas it increased steadily in the zygotic embryos. The fatty acid composition of individual cotyledons from microspore embryos shake cultured for 15 days was compared with that of individual mature seeds. Relative amounts of the major fatty acids, i.e. palmitic, oleic and linoleic acids, were essentially the same, whereas the microspore-derived embryos had about 35% less stearic acid and 35% more linolenic acid than the mature seeds. Variation in the amounts of oleic, linoleic and linolenic acids between seeds was similar to that found between cotyledons of microspore-derived embryos, whereas variation in palmitic and stearic acid levels was significantly lower between microsporederived cotyledons than between the seeds. The results indicate that microspore-derived embryos from shake cultures should be convenient for use in studying the regulation of oil biosynthesis and for rapidly screening for oil quality in genetically altered rapeseed.