Solubilization and partial purification of protein kinase systems from brain membranes that phosphorylate calspectin: A spectrin-like calmodulin-binding protein (fodrin)

In brain tissue a spectrin-like calmodulin-binding protein calspectin, or fodrin, is concentrated in a synaptosome fraction, where most of the calspectin is associated with the synaptic membranes. This endogenous calspectin was phosphorylated by protein kinase system(s) associated with the membranes. Here, we report the solubilization and partial purification of the membrane-associated calspectin kinase activity. The activity was resolved on a gel filtration column into two fractions, peaks I and II having estimated M_r of 800 000 and 88 000. The activity of peak I was dependent on the presence of both Ca²⁺ and calmodulin. Peak II revealed a basal activity in the absence of Ca²⁺ and calmodulin, which was stimulated 2-fold by addition of Ca²⁺. Calmodulin had no effect on the peak II activity.     

Relative addition rate and external concentration; Driving variables used in plant nutrition research

Abstract An increasing literature accounting for various types of experiments indicates that far lower external nutrient concentrations are required by plants than is usually thought to be the case. It is concluded that the ion uptake capacity of the roots, as described by the carrier concept, is high compared to that required for maintenance of the internal concentration. Serious errors in experimental conclusions are associated with insufficient and constant nutrient addition rates. The main errors are caused by non-steady states of the plants both with regard to the internal nutrient concentrations and the relative growth rate. A dynamic concept has been proposed for direct use as the treatment variable within the range of sub-optimum nutrition. The nutritional factor is expressed as a flow, the relative nutrient addition rate in laboratory studies and the nutrient flux density in the field. The experimental use of the relative addition rate has led to steady-state nutrient status and relative growth rate and the interpretation of plant responses which differ fundamentally from accepted views. Thus, for instance, deficiency symptoms disappear, as in natural conditions, when the internal nitrogen concentration is stable, independent of level. The nutrition/growth relationships are very different from those observed when external concentration is varied. The regression line of relative growth rate on relative addition rate passes near to the origin at an angle close to 45 to the axes, which implies that the obtained relative growth rate approximates closely the treatment variable. A striking example of observed differences is the positive effect on nitrogen fixation exerted by high relative nitrogen addition rates compared to the well-known negative effect of increasing external nitrogen concentration. The application of fertilizer on the basis of the nutrient flux density concept provides the possibility of supplying fertilizers corresponding to the consumption potential of the vegetation and to the natural flux density resulting from mineralization in the soil. Nitrogen utilization is high under such conditions and the resulting feedback of nutrition on the mineralization rate suggests that there will be a long-term increase in fertility.     

Dérivés d'acides 3-(β-d-érythrofuranosyl)-pyrazolecarboxyliques

Treatment of 2,5-anhydro-1-bromo-1-deoxy-2,3-O-isopropylidene-1-p-nitrophenylhydrazono-d-ribose with methyl acetylenecarboxylate gave methyl 3-(2,3-O-isopropylidene-β-d-erythrofuranosyl)-1-p-nitrophenylpyrazole- (8) and 5-carboxylate (9). Amidification at C-5 of 8 was easier than at C-4 of 9. Similarly, dimethyl 3-(2,3-O-isopropylidene-β-d-erythrofuranosyl)- 1 - p-nitrophenylpyrazole-4,5-dicarboxylate gave specifically a 5-carbamoyl derivative, the structure of which was established by comparison of the ¹³C-n.m.r.spectrum with those of a series of glycosylpyrazoles. The correlation between the experimental values of the chemical shifts of the carbon atoms of the pyrazole ring and the values calculated by addition of the contributions of the various groups linked to the ring was better (R 0.98) than the correlations obtained by calculation by the CNDO/2 method of the total electron population (R 0.92) or of the π-electron population of each carbon atom (R 0.85).     

Effects of nitrogen supply on the anatomy and chemical composition of leaves of four grass species belonging to the genus Poa, as determined by image-processing analysis and pyrolysis–mass spectrometry

Previous experiments have shown that the anatomy and chemical composition of leaves of inherently fast- and slow-growing grass species, grown at non-limiting nitrogen supply, differ systematically. The present experiment was carried out to investigate whether these differences persist when the plants are grown at an intermediate or a very low nitrogen supply. To this end, the inherently fast-growing Poa annua L. and Poa trivialis L., and the inherently slow-growing Poa compressa L. and Poa pratensis (L.) Schreb. were grown hydroponically at three levels of nitrate supply: at optimum (RGR_max) and at relative addition rates of 100 and 50 mmol N (mol N)^?1 d^?1 (RAR₁₀₀ and RAR₅₀), respectively. As expected, at the lowest N supply, the potentially fast-growing species grew at the same rate as the inherently slow-growing ones. Similarly, the differences in leaf area ratio (LAR, leaf area:total dry mass), specific leaf area (SLA, leaf arear:leaf dry mass) and leaf mass ratio (LMR, leaf dry mass:total dry mass) disappeared. Under optimal conditions, the fast-growing species differed from the slow-growing ones in that they had a higher N concentration. There were no significant differences in C concentration. With decreasing N supply, the total N concentration decreased and the differences between the species disappeared. The total C concentration increased for the fast-growing species and decreased for the slow-growing ones, i.e. the small, but insignificant, difference in C concentration between the species at RGR_max increased with decreasing N supply. The chemical composition of the leaves at low N supply, analysed in more detail by pyrolysis–mass spectrometry, showed an increase in the relative amounts of guaiacyl lignin, cellulose and hemicellulose, whereas those of syringyl lignin and protein decreased. The anatomy and morphology of the leaves of the four grass species differing in RGR_max were analysed by image-processing analysis. The proportion of the total volume occupied by mesophyll plus intercellular spaces and epidermis did not correlate with the amount of leaf mass per unit leaf area (specific leaf mass, SLM) at different N supply. The higher SLM at low N supply was caused partly by a high proportion of non-veinal sclerenchymatic cells per cross-section and partly by the smaller volume of epidermal cells. We conclude that the decrease in relative growth rate (and increase in SLM) at decreasing N supply is partly due to chemical and anatomical changes. The differences between the fast- and slow-growing grass species at an optimum nutrient supply diminished when plants were growing at a limiting nitrogen supply.     

The identification of 7-[(R)-2-((1S,2S)-2-benzyloxycyclopentylamino)-1-hydroxyethyl]-4-hydroxybenzothiazolone as an inhaled long-acting β2-adrenoceptor agonist

The optimisation of two series of 4-hydroxybenzothiazolone derived β₂-adrenoceptor agonists, bearing α-substituted cyclopentyl and β-phenethyl amino-substituents, as inhaled long-acting bronchodilators is described. Analogues were selected for synthesis using a lipophilicity based hypothesis to achieve the targeted rapid onset of action in combination with a long duration of action. The profiling of the two series led to identification of the α-substituted cyclopentyl analogue 2 as the optimal compound with a comparable profile to the inhaled once-daily long-acting β₂-adrenoceptor agonist indacaterol. On the basis of these data 2 was promoted as the backup development candidate to indacaterol from the Novartis LABA project.     

On the Information Matrix of Exponential Mixture Models with Long-term Survivors

The aim of this paper is to study the properties of the asymptotic variances of the maximum likelihood estimators of the parameters of the exponential mixture model with long-term survivors for randomly censored data. In addition, we study the asymptotic relative efficiency of these estimators versus those which would be obtained with complete follow-up. It is shown that fixed censoring at time T produces higher precision as well as higher asymptotic relative efficiency than those obtainable under uniform and uniform-exponential censoring distributions over (0, T). The results are useful in planning the size and duration of survival experiments with long-term survivors under random censoring schemes.