Production of δ-(l-α-aminoadipyl)-l-cysteinyl-d-valine by entrapped ACV-synthetase fromStreptomyces clavuligerus

Summary -(l--Aminoadipyl)-l-cysteinyl-d-valine (ACV)-synthetase fromStreptomyces clavuligerus was studied under conditions that enabled the reuse of the enzyme. Coupling of ACV-synthetase to DEAE-Trisacryl and aminopropyl-glass resulted in an immobilized enzyme product of little or no catalytic activity. However, an enzyme reactor was designed by physical confinement of partially-purified ACV-synthetase in an ultrafiltration cell. This system was stimulated by phosphoenolpyruvate at lower concentrations of ATP, an effect not observed with purified enzyme. Up to 30% conversion of the limiting substrate, cysteine, to ACV occurred under semi-continuous conditions. Reaction products were investigated as potential inhibitors: AMP was the most inhibitory, but only when used at concentrations in excess of those produced in reaction mixtures. Under a nitrogen atmosphere, both product and enzyme stabilities were greatly improved and the enzyme retained 45–46% of its initial activity after five uses at room temperature during a 24-h period. Extrapolations based on these data suggest that 1.3 g partially purified enzyme (0.13 U g^–1) would be capable of producing 411 mg of ACV in a 1-L reaction mixture in this period.     

Some improvements in the quality of NAA procedures and the reliability of the results

After considering the need for quality control in NAA, the concept of quality in NAA procedures themselves is discussed, and some important factors identified. Two approaches to improve quality are then described in more detail. The first concerns the unique ability of NAA using different isotopic reactions and different modes (INAA/RNAA) to provide independent data sets in the same laboratory, thus allowing internal validation or crosschecking. The second discusses the need for chemical yield measurements in RNAA and the advantages of the radioisotopic tracer technique. Some recent advances and further possibilities for this use of tracers are listed.     

Uptake,accumulation and metabolism of auxins in tobacco leaf protoplasts

Uptake and metabolism of exogenous naphthalene-1-acetic acid (NAA) and indole-3-acetic acid (IAA) have been studied in tobacco (Nicotiana tabacum L. cv. Xanthi) mesophyll protoplasts. Both auxins entered protoplasts by diffusion under the action of the transmembrane pH gradient without any detectable participation of an influx carrier. Molecules were accumulated by an anion-trapping mechanism and most of them were metabolized within hours, essentially as glucose-ester and amino-acid conjugates. Protoplasts were equipped with a functional auxin-efflux carrier as evidenced by the inhibitory effect of naphthylphtalamic acid on IAA efflux. Basically, similar mechanisms of NAA and IAA uptake occurred in protoplasts. However, the two auxins differed in their levels of accumulation, due to different membrane-transport characteristics, and the nature of the metabolites produced. This shows the need to estimate the accumulation and the metabolism of auxins when analyzing their effects in a given cell system. The internal auxin concentration could be modulated by changing the transmembrane pH gradient, giving an interesting perspective for discriminating between the effects of intra- and extracellular auxin on physiological processes.Abbreviations BA benzoic acid - C_i/C_e accumulation ratio of auxin - IAAasp N-[3-indolylacetyl]-dl-aspartic acid - NAA naphthalene-1-acetic acid - NAAasp N-[1-naphthylacetyl]-l-aspartic acid - NPA N-1-naphthylphthalamic acid The authors thank Dr. M. Caboche (I.N.R.A, Versailles, France) for his generous gifts of some amide derivatives of 1-NAA, Mr. P. Varennes and Dr. B. Das (I.C.S.N., C.N.R.S., Gif-sur-Yvette, France) for recording and interpreting the mass spectra of NAA glucose ester, and Prof. P. Manigault (Institut des Sciences Végétales, Gif-sur-Yvette) for microscopy measurements of protoplast dimensions. This work was supported by funds from the C.N.R.S, I.N.R.A, and E.E.C.     

Iron toxicity and other chemical soil constraints to rice in highland swamps of Burundi

Iron toxicity is suspected to be a major nutritional disorder in rice cropping systems established on flooded organic soils that contain reductible iron. A pot trial was carried out to assess Fe toxicity to rice in flooded Burundi highland swamp soils with a wide range of organic carbon contents. Soil and leaf analyses were performed and total grain weight was determined. Clear Fe toxicity was diagnosed, based on leaf Fe content at panicle differentiation. Leaf Fe contents higher than 250 g g^–1 dry matter induced lower Mg (and probably Mn) uptake, and a 50% total grain weight reduction. These features were associated with exchangeable Fe equivalent fractions higher than 86%. Besides, several non-Fe toxic soils exhibited an Mg-Mn imbalance.     

Simple electromyographic biofeedback treatment for chronic pediatric constipation/encopresis: Preliminary report

Pediatric constipation/encopresis is thought to be due, in part, to paradoxical constriction of the external anal sphincter (EAS) muscle during attempted defecation. This inappropriate contraction can lead to delayed, impacted, painful, and infrequent bowel movements. Standard Medical Care (SMC) involves disimpaction with enemas, followed by laxative therapy and diet modification, to maintain frequent soft stools. Using the case control method, the efficacy of SMC alone was compared with SMC plus EAS electromyographic biofeedback aimed at eliminating paradoxical contraction. Thirteen consecutive chronically constipated children received SMC plus biofeedback, and were compared with 13 age- and sex-matched children who received only SMC. Biofeedback subjects demonstrated post-treatment elimination of EAS paradoxical constriction. At 16 months follow-up parents of biofeedback children reported significantly greater improvement in constipation, encopresis, laxative use, and painful bowel movements compared to SMC.This research report was supported by the NIH under grant No. RO1 HD 28160.     

High-temperature gas chromatography and gas chromatography-mass spectrometry of glycoprotein and glycosphingolipid oligosaccharides

High-temperature gas chromatography and gas chromatography-inass spectrometry for the analyses of oligosaccharides derived from glycoproteins or glycosphingolipids has been developed. Pcrmethylatcd oligosaccharides with up to about 12 sugar residues and masses up to 2500 Daltons can be analyzed. This approach is discussed and exemplified.     

Evolution of aging: Testing the theory usingDrosophila

Evolutionary explanations of aging (or senescence) fall into two classes. First, organisms might have evolved the optimal life history, in which survival and fertility late in life are sacrificed for the sake of early reproduction or high pre-adult survival. Second, the life history might be depressed below this optimal compromise by the influx of deleterious mutations; since selection against late-acting mutations is weaker, deleterious mutations will impose a greater load on late life. We discuss ways in which these theories might be investigated and distinguished, with reference to experimental work withDrosophila. While genetic correlations between life history traits determine the immediate response to selection, they are hard to measure, and may not reflect the fundamental constraints on life history. Long term selection experiments are more likely to be informative. The third approach of using experimental manipulations suffers from some of the same problems as measures of genetic correlations; however, these two approaches may be fruitful when used together. The experimental results so far suggest that aging inDrosophila has evolved in part as a consequence of selection for an optimal life history, and in part as a result of accumulation of predominantly late-acting deleterious mutations. Quantification of these effects presents a major challenge for the future.     

Vegetative growth and productivity ofEichhornia azurea with special emphasis on leaf dynamics

The lifeform and the biological production of pure stands ofEichhornia azurea Kunth in three lakes in tropical Brazil were studied. The lifeform ofE. azurea is termed ‘semi-emergent’, because the plant has well developed trailing stems just under the water, and the aerial lamina emerges with the thick petiole. The density of shoot apices was 9.9, 17.2 and 17.1 m⁻² in Lake Dom Helvecio, Lake Jacaré and Lake Carioca, respectively. The mean daily increment of the apical shoot biomass was between 1.8 and 4.8 g m⁻² day⁻¹. The mean leaf life-span in Lake Dom Helvecio, Lake Jacaré and Lake Carioca was estimated to be 78, 49 and 64 days in the wet season and 73, 70 and 73 days in the dry season, respectively. The stem life-span was estimated to be about 28 months. Starch content in the current years' stem ranged from 24 to 118 mg g⁻¹ dry matter with fluctuations, the amplitude of which decreased with age. The differences for most of the growth parameters, such as density of shoot apices, daily increment of biomass and leaf life-span, between dry and wet season are smaller than those among the three lakes. Both the decrease in daily dry matter production and the increase in leaf life-span occurred in order from Lake Dom Helvecio to Lake Jacaré and Lake Carioca. The low productivity ofE. azurea is considered to be related to a low leaf area index, a long time interval for the emergence of new leaves, long leaf life-span and a low capacity for branching.     

SEASONAL PATTERNS OF MORPHOLOGICAL VARIABILITY IN SARGASSUM POLYCERATIUM (PHAEOPHYTA)1

Temporal variability in certain morphological and taxonomically important features was quantified for Sargassum polyceratium Mont. from a population in the Content Keys, Florida (U.S.A.). Patterns of blade development, senescence, and loss caused pronounced seasonal changes in blade length-width ratios. Blade length and width were maximal early in the growing season (August-November) and decreased as the annual stems matured. Early in the growing season, plants had broader blades with randomly distributed cryptostomata. Late in the growing season (February-April), plants had more linear blades with cryptostomata approximately arranged in two rows, one on each side of the midrib. The length-width ratio of blades increased acropetally along the stems and were directly correlated to the size of the cryptostomatal opening and inversely correlated with the number of cryptostomata. The branching pattern of the annual stems ranged from short spur branches to well-developed, lateral axillary branches. The frequency of bifurcated blades increased significantly late in the growing season. Vesicle shape and size and pedicel length were temporally stable. Alated pedicels and mucronate vesicles occurred in low frequencies. The variability of the morphological features used to delineate species within the genus Sargassum on the tropical eastern coasts of the Americas is poorly understood.     

Abundance of phytoseiid mites on Vitis species: effects of leaf hairs,domatia, prey abundance and plant phylogeny

We observed the number of predatory mites (Phytoseiidae:Typhlodromus caudiglans) on the foliage of 20 North American species of grapes (Vitis spp) plus the domesticated EuropeanVitis vinifera, all grown in a common garden. We found relatively few phytophagous mites. The numbers of phytophagous mites were not correlated with the plant characteristics that we measured. We found approximately five times as many predatory mites as phytophagous mites and the numbers of these phytoseiid predators were not affected by the availability of prey. Similarly, numbers of phytoseiids were unaffected by plant gender and, hence, the availability of pollen, another source of food. The numbers of phytoseiids were not clustered according to the taxonomic grouping of the tested plant species. Leaf surface characteristics explained over 25% of the variance in the numbers of phytoseiids. Numbers of phytoseiids were positively associated with the density of vein hairs, the density of bristles in leaf axils, and the presence of leaf domatia. These results suggest that sheltered habitats rather than food availability may limit the numbers of phytoseiid mites on grapevines.