Calcium-stimulated myofibrillar ATPase activity correlates with shortening velocity of muscle fibres in Xenopus laevis

Summary The iliofibularis muscle ofXenopus laevis is reported to contain five types of fibres which have different force—velocity relationships. Ten fibres of each type were selected on the basis of succinate dehydrogenase activity, cross-sectional area and location in the muscle, in order to assess the validity of the fibre type classification.Maximum calcium-stimulated myofibrillar ATPase activity (V _max) and apparent Michaelis constant (K _m) for ATP were determined for these 50 fibres from serial sections. The values obtained varied according to the type of fibre. Type 1 had the highest and type 5 the lowest values forK _m andV _max.In a separate experiment, single freeze-dried fibres were used to determine the relationship between their ATP content and apparentK _m for ATP. There was a tendency for high ATP concentrations in fibres with highK _m values.When myofibrillar ATPase activity was related to the maximum velocity of shortening of the five fibre types, a significant correlation was found. It is concluded that calcium-stimulated myofibrillar ATPase histochemistry allows an estimate of the maximum shortening velocity of muscle fibres fromXenopus laevis.     

Use of commercially available monoclonal antibodies for immunoenzyme double staining

Summary An immunoenzyme double-staining method for the simultaneous detection of two cellular epitopes, using commercially available mouse monoclonal antibodies, is described. The method employs a combination of the suppression of endogenous biotin and two successive indirect techniques with a blocking step in between. The first indirect method involves an unlabelled monoclonal antibody followed by an alkaline phosphatase-conjugated goat anti-mouse immunoglobulin. After a blocking step with normal mouse serum, the second indirect method is applied using a biotinylated monoclonal antibody followed by the visualization of this antibody by avidin-biotinylated peroxidase complex (ABC) or rabbit anti-biotin and peroxidase-conjugated swine anti-rabbit immunoglobulin in successive steps. Using these methods in combination with the introduction of dioctyl sodium sulphosuccinate and tetramethylbenzidine as chromogens for peroxidase activity, two cellular epitopes could be distinguished clearly in tissue sections by the green-and violet-stained peroxidase and alkaline phosphatase activities, respectively. The expression of two epitopes on the same cellular constituent is outlined by the coappearance of both enzyme activities as a bluish-purple colour. This method allows for the simultaneous identification, localization and enumeration of two cellular epitopes. These can serve as parameters for a number of pathological processes.     

Candida delta-aminovalerate: alpha-ketoglutarate aminotransferase: purification and enzymologic properties

A new enzyme that catalyzes the transamination of delta-aminovalerate with alpha-ketoglutarate was purified to homogeneity from adapted cells of Candida guilliermondii var. membranaefaciens. The relative molecular mass determined by gel filtration was estimated to be close to 118,000. The transaminase behaved as a dimer with two similar subunits in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme has a maximum activity in the pH range of 7.8-8.5 and at 40 degrees C. alpha-Ketoglutarate and to a lesser extent pyridoxal 5'-phosphate were effective protecting agents toward temperature raising. The enzyme exhibits absorption maximum at 330 and 410 nm. The enzyme catalyzes the transamination between omega-amino acids and alpha-ketoglutarate. delta-Aminovaleric acid is the best amino donor. The Km values for delta-aminovalerate, alpha-ketoglutarate, and pyridoxal 5'-phosphate determined from the Lineweaver-Burk plot were 4.9 mM, 3.6 mM, and 22.7 microM, respectively. The inhibitory effect of various amino acids analogues on the transamination reaction between delta-aminovalerate and alpha-ketoglutarate was studied, and Ki values were determined.     

Classical Analysis of Variance Methods and Nonparametric Counterparts

The present investigation is concerned with the use of classical and nonparametric techniques in the analysis of experiments. Occasionally biometricians are confronted with results arising from an experiment with data which do not necessarily satisfy the assumption of Normality, the basis of the classical analysis of variance methods. Sometimes the biometrician handles such a situation by transformation of the observations and applying the classical techniques. Although this attack may lead to a satisfactory analysis, the results may be questionable in a number of cases. In analyzing continuous data without the Normality assumption nonparametric methods provide realistic alternatives. In this paper a number of problems will be discussed with and without the Normality assumption resulting in the well-known classical analysis and its nonparametric counterparts. Most of the nonparametric procedures to be discussed are based on ranks.     

Initial decomposition ofNymphoides peltata (Gmel.) O. Kuntze (Menyanthaceae), as studied by the leaf-marking method

Summary In this paper the leaf-marking method as used for the study of the development and initial decomposition of floating leaves is described and the reliability of the various measurements is tested and/or discussed. Some general results obtained withNymphoides peltata (Gmel.) O. Kuntze in tanks and in the field are presented and crltically discussed. Autolysis followed by microbial decay was in all cases the most important factor by which leaves disintegrated. In the field plots animals were responsible for the disappearance of 22% of the total leaf area produced during a growth season. This is, however, the combined effect of consumption and damage succeeded by microbial decay. Real grazing can be estimated to be no more than 10% of the production of floating leaves. Fungi can have an important role in initial decomposition, especially after the flowering period, as is demonstrated forSeptoria villarsiae Desm. All damage types show temporal and, in the case of animals, also spatial distribution patterns.     

A caterpillar (Callopistria floridensis G. (Lepidoptera: Noctuidae)) accumulates arsenic from an arsenic‐hyperaccumulating fern (Pteris vittata L.).

1. The consumption of arsenic is toxic to most biota. However, a noctuid caterpillar was recently reported feeding on a plant known to hyperaccumulate arsenic. 2. The aim of this study was to investigate the effects of arsenic‐rich Pteris vittata L. consumption by Callopistria floridensis G., and measure differences in arsenic concentrations at various stages of development (larval and adult), and associated with exuviae and frass. 3. Callopistria floridensis accumulated extraordinary concentrations of arsenic. The relative accumulation of arsenic was highest in exuviae and larvae. Larvae invariably preferred P. vittata grown on low arsenic soil to P. vittata grown on higher soil arsenic concentrations, and appeared able to selectively forage on lower arsenic concentrations within each treatment. 4. These findings show that C. floridensis is tolerant of arsenic, and successfully develops to adulthood containing elevated concentrations of arsenic. Callopistria floridensis represents the only known terrestrial animal capable of accumulating arsenic, and may have developed novel physiological and behavioural adaptations to regulate the negative effects of arsenic.