Analysis of pipecolic acid in biological fluids using capillary gas chromatography with electron-capture detection and [H11]pipecolic acid as internal standard

A sensitive and accurate stable isotope dilution assay was developed for the measurement of pipecolic acid in body fluids using capillary gas chromatography with electron-capture detection. The method utilizes [²H₁₁]pipecolic acid as the internal standard. Sample preparation consisted of derivatization in aqueous solution (pH 11.5) of the amine moiety with methyl chloroformate to the N-methylcarbamate, followed by acidic ethyl acetate extraction at pH ≤ 2 and further derivatization of the carboxyl moiety with pentafluorobenzyl bromide, the excess of which was removed by solid-phase extraction. Control values have been determined in the plasma of at-term infants, age > 1 week (n = 21, mean = 1.36 μM, range = 0.47–3.27 μM). The utility of the method was demonstrated by quantitating pipecolic acid in biological fluids derived from patients with peroxisomal disorders. The method was validated against an established electron-capture negative ion mass fragmentographic technique.     

Primary structure of pyruvate, orthophosphate dikinase in the dicotyledonous C4 plant Flaveria trinervia

We have isolated and characterized cDNA clones encoding the entire precursor for the leafspecific isoform of pyruvate, orthophosphate dikinase (PPDK) from the dicotyledonous C4 plant Flaveria trinervia. The deduced amino acid sequence reveals a high degree of similarity to the corresponding maize protein indicating a common evolutionary basis. However, no significant similarities are apparent upon comparison of the putative transit peptides. The implications of this divergence are discussed with respect to the evolution of PPDK genes.     

Correlation between adult transformation and aldehyde oxidase staining pattern in wing discs ofApterous genotypes inDrosophila melanogaster

Summary The aldehyde oxidase staining pattern in wing discs ofDrosophila melanogaster bearing the genotypesap ^blt /ap ^blt andap ^blt andap ^blt /ap ⁷³ⁿ showns changes from the wild-type pattern. Extensive areas of the presumptive dorsal posterior wing blade, which are normally unstained, have enzyme activity in these mutants. In wings of these genotypes, dorsal posterior structures are replaced by dorsal anterior wing structures. A strong correlation has been found between the frequencies of various staining patterns in the discs and the extent of transformation in the cuticular structures of the wing, which is consistent with the idea that aldehyde oxidase activity can be used as an indicator in the wing disc of this transformation. Unlike the homoeotic mutationengrailed, apterous has not been interpreted as a selector gene yet the work reported here shows thatapterous alleles can cause changes resembling those of theengrailed phenotype both in aldehyde oxidase staining behaviour and in the cuticular transformation.     

The Receptor-Mediated Activation of Tyrosine Hydroxylation in the Superior Cervical Ganglion of the Rat

The addition of carbachol to superior cervical ganglia causes a rapid increase in tyrosine hydroxylation in situ. The increase occurs in ganglia from both newborn and adult animals, and in ganglia from animals pretreated with reserpine. The increase is not due to increased transport of the substrate. The increase is dependent upon the presence of calcium, and is additive to the stimulation produced by dibutyryl cyclic AMP. The stimulation seems specific for tyrosine hydroxylation; dopamine beta-hydroxylation is not increased. Preincubation experiments suggest that the carbachol-induced stimulation is due to a change in the availability of, or the affinity of the enzyme for, reduced pterin cofactor. The stimulation is inhibited by atropine and also by low concentrations of phenoxybenzamine or haloperidol, which suggests that it is caused by an action of carbachol on the interneurons in the ganglia.     

Diterpene carboxylic acids and a heliangolide from Helianthus angustifolius

Two diterpene carboxylic acids, one a new kaurenoid derivative and one the previously characterized labdane, ()-cis-ozic acid, as well as a     

Stratigraphic occurrences and vegetational patterns of Pennsylvanian pteridosperms in Euramerican coal swamps

The vegetation of many Euramerican coal swamps is known from coal-ball peats in the tropical rainy zone across the paleocontinent of Laurussia in the Upper Carboniferous. The stratigraphic occurrences of coal balls in Europe (lower Westphalian) and in North America (upper Westphalian and lower Stephanian) are largely complementary and a revised correlation chart is given. Quantitative analyses of the vegetation from these autochthonous peats are combined with coal palynology to determine on overview of swamp vegetation patterns during the Westphalian and early Stephanian. In comparison to the dominance and great diversity of pteridosperms in the Upper Carboniferous compression floras, seed ferns were minor to subdominant elements in the coal-ball peats. The quantitative composition of organ assemblages of the principal pteridosperms is given with stratigraphic occurrences of Medullosa and Sutcliffia (Medullosaceae), Heterangium, Lyginopteris, Microspermopteris, and Schopfiastrum (Lyginopteridaceaea) and Callistophyton (Callistophytaceae). The major peat contributors among pteridosperms are Lyginopteris (?90%) in the Westphalian A and Medullosa (?95%) in the Westphalian D and Stephanian. Lyginopteris became extinct in the early Westphalian B and Medullosa became abundant during the Westphalian C–D transition and a subdominant in the Stephanian contributing 13–21% of the permineralized peats. Paleoecological interpretations indicate that medullosan trees were most abundant but patchy in distribution in drier swamp stages with enriched nutrients and near major channels in swamps during the Westphalian D. It is suggested that the swamp pteridosperms are probably quite similar to or identical with some of the representatives in the more diverse compression floras. The evolution and diversity noted in swamp pteridosperms are considered principally the product of adjacent lowland communities which repeatedly introduced seeds into the wetlands. The swamp pteridosperms exhibit the greatest fluctuation in abundance (biomass) from site to site in the same coal, probably the highest level of diversity and the only steady pattern of increase in importance during the Late Carboniferous with the possible exception of the Westphalian B–C interval. The close relationships inferred between swamp pteridosperms and their lowland seed sources are also compatible with the frequent association of tree ferns and seed ferns during the Westphalian D and in the Stephanian when these two kinds of frond-bearing trees formed most of the vegetation in both lowland and swamp environments.     

Carbohydrate metabolism during cold-induced sweetening of potato tubers

The aim of this work was to discover the effects of lowering the temperature from 25° to 2° on the metabolism of glucose [U-¹⁴C] by tubers of Solanum tuberosum. Isotope was applied to tubers via a 50-μl hole made with a capillary pipette. Tubers were incubated for 2 hr, the pulse; then the glucose- [U-¹⁴C] was replaced with glucose, and incubation was continued for 18 hr, the chase. The detailed distribution of ¹⁴C was determined at the end of the pulse and at the end of the chase at 2°, and compared with those found at 25°. Lowering the temperature reduced the proportion of metabolized ¹⁴C that entered the respiratory pathways. At 2°, but not at 25°, hexose phosphates were the most heavily labelled fraction after the pulse: during the chase at 2° much of this label was metabolized to sucrose. We conclude that lowering the temperature preferentially restricts glycolysis and diverts hexose phosphates to sucrose. We suggest that this is an important cause of cold-inducing sweetening of the tubers and is due to cold-lability of key glycolytic enzymes.     

Use of the smith degradation in the study of the branching pattern in the complex-type carbohydrate units of glycoproteins

The site of substitution of the peripheral branches on the core mannose-residues in the complex-type carbohydrate units of glycoproteins has been studied by Smith degradation. By using glycopeptides of known structure, it is shown that the site of attachment of the (1→4)-linked, third branch to the mannose core in tri- and tetra-antennary structures may be conveniently demonstrated by this technique. A critical factor was found to be the concentration of acid used for cleavage of the periodate-oxidized product. This finding could explain discrepancies concerning the previously published structures of fetuin glycopeptides. The technique was applied for study of the branching pattern in human transferrin, porcine thyroglobulin, and rat plasma and brain glycoproteins. The results support the concept that, in glycoproteins from most sources, the third (1→4)-linked branch is attached to the (1→3)-linked mannose residue of the core portion in the complex-type structures. An indication for a novel type of branching pattern in human transferrin was, however, also obtained.     

Induction of tyrosine aminotransferase by pyridoxine in Drosophila hydei

Salivary glands incubated in various concentrations of pyridoxine (Vitamin B₆) show increasing tyrosine aminotransferase (TAT) activity at concentrations up to 10^–5 M and then decreasing activity up to 10^–2 M but in all cases the activity is greater than that of the controls. This increase in activity is demonstrable for up to 6 h, the longest period tested, and is dependent on the synthesis of new mRNA. A similar increase in TAT activity is observed in salivary glands subjected to heat shock. Antibodies prepared against purified tyrosine aminotransferase precipitate a peptide of the same molecular weight (40 KD) as that induced by pyridoxine.     

A 13c-n.m.r. study of 1,6:2,3- and 1,6:3,4-dianhydro-β-D-hexo-pyranoses and their O-acetyl and deoxy derivatives

¹³C-N.m.r. spectra of all possible 1,6:2,3- and 1,6:3,4-dianhydro-β-D-hexo-pyranoses and their O-acetyl and deoxy derivatives are presented. Relations between chemical shifts of certain carbon atoms and the structure of the dianhydrides are outlined, and their application in structural analysis is discussed. Inversion of configuration of the oxirane ring from the endo to the exo position is associated with typical upfield-shifts for oxirane-ring carbon atoms C-2 or C-4, respectively. Possible inter-relationships between ¹³C-chemical shifts and steric and polar interactions in the dianhydro derivatives are discussed.