Degradation of coniferyl alcohol and other lignin-related aromatic compounds by Nocardia sp. DSM 1069

Nocardia sp. DSM 1069 was grown on mineral salt media with coniferyl alcohol, 4-methoxybenzoic acid, 3-methoxybenzoic acid or veratric acid as sole sources of carbon and energy. During incubation on coniferyl alcohol, the formation of coniferyl aldehyde, ferulic acid and quantitative accumulation of vanillic acid and proteocatechuic acid could be achieved with mutants. Washed cell suspensions of N. sp. grown on 4-methoxybenzoic acid, oxidized 4-hydroxybenzoic acid and protocatechuic acid. Cells grown on veratric acid, oxidized vanillic acid, isovanillic acid, and protocatechuic acid. Cell extracts were shown to cleave protocatechuic acid by ortho-fission.A mutant without protocatechuate 3,4-dioxygenase activity was not influenced in its growth on 3 methoxybenzoic acid. Cell free extracts of cells grown on 3-methoxybenzoic acid were shown to catalyze the oxidation of gentisic acid (2,5-dihydroxybenzoic acid). The resulting ring cleavage product was further metabolized by a glutathione dependent reaction.The specificity of the demethylation reactions has been investigated with a mutant unable to grow on vanillic acid. This mutant was not impaired in the degradation of isovanillic acid, 4-methoxy-, or 3-methoxybenzoic acid, whereas growth of this mutant on veratric acid (3,4-dimethoxybenzoic acid) was only half as much as that of the wild type. Concomitantly with growth on veratric acid this mutant accumulated vanillic acid with a yield of about 50%.A pathway for the catabolism of coniferyl alcohol, involving oxidation and shortening of the side chain, and of 4-methoxybenzoic acid and veratric acid with protocatechuic acid as intermediate is being proposed. A second one is proposed for the degradation of 3-methoxybenzoic acid with gentisic acid as intermediate.     

Two types of luliberin-immunoreactive perikarya in the preoptic area of the rat

Summary At the light microscopic level, following immunostaining with a single antiserum against luliberin (LRF), two types of hormone-producing perikarya in the preoptic area are demonstrated. The two cell types differ in their morphological features: a bipolar, smooth-contoured cell type can be differentiated from an irregularly contoured unipolar type. Intermediate forms between both cell types occurring in the same area are not observed. Electron microscopically, both cell types contain labeled granules of similar size and immunoreactivity. It is dicussed whether the uneven surface of the one cell type is due to areas of synaptic contacts, and whether both cell types are integrated in different neuronal and functional circuits. Moreover, at the ultrastructural level, from the irregularly contoured LRF-producing perikarya a further positively stained cell type, probably a glial cell, can be differentiated. The specific labeling of the latter is caused by its content of immunoreactive lysosomal bodies. Differentiation between the labeled glial cells and the irregularly contoured LRF-producing perikarya is not possible at the light microscopic level.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/3) and by the Stiftung Volkswagenwerk     

Presence of adrenocorticotropin and β-Endorphin immunoreactivities in the magnocellular neurosecretory system of the rat hypothalamus

Summary Antisera raised against ACTH (1–39), -endorphin and the 16K proopiocortin were used, in association with the immunoperoxidase reaction, to localize positively-staining cell bodies and nerve fibres in the hypothalamus of the rat. Antigens, cross-reactive against anti-ACTH (1–39) serum were detected in a fibre system in the rostro-dorsal hypothalamus situated between the optic chiasm and the third ventricle while immunoreactive 16K-like material was present in fibres localized in the caudal hypothalamus, dorso-lateral to the arcuate nucleus. This latter system was also associated with the appearance of ACTH (1–39) and ACTH (17–39) immunoreactivity.Cells of the arcuate nucleus stained positively for ACTH (1–39), 16K antigen and -endorphin, and on examining adjacent thin sections it was observed that cells that contained 16K antigen-like material, also gave a positive immunoreaction with ACTH (1–39) and -endorphin antisera. In the magnocellular system, cells of the supraoptic (SON) and paraventricular (PVN) nuclei also gave a positive immunoreaction with anti-ACTH (1–39), 16K antigen and -endorphin serum. As in the case of the arcuate nucleus, common cells stained for these three antigens.On the basis of the precursor theory for the synthesis of ACTH, 16K antigen and -endorphin, it was not unexpected to find these three fragments of pro-opiocortin localized together in cells of the arcuate nucleus. That ACTH (1–39), 16K antigen and -endorphin-like materials are present in the magnocellular neurosecretory system would suggest that cells of the SON and PVN are not only involved in the synthesis of neurophysin and the neurohypophysial hormones, but also of some products of the pro-opiocortin molecule. Whether the biochemical nature of the ACTH and -endorphin in cells of the SON and PVN is identical to that of anterior pituitary origin remains to be established, as does the biosynthetic relationship between neurophysin and oxytocin/ vasopressin and these fragments of pro-opiocortin.Drs. M.M. Wilkes, S.S.C. Yen, G. Pelletier, B.A. Eipper and R. Walter are thanked for supplying some of the antisera and antigens used in this study. Thanks also go to Ciba-Geigy Ltd. and Organon Inc. for supplies of ACTH (17–39) and ACTH (1–24) respectively. This work was financed by The Medical Research Council of New Zealand     

Inhibitory effects of different quantitative compositions of Hymenaea leaf resins on a generalist herbivore Spodoptera exigua

The hypotheses that quantitative variation in leaf resins in the leguminous genus Hymenaea may partially be a response to insect predation was tested in feeding experiments with the generalist herbivore, the beet armyworm, Spodoptera exigua Hübn (Noctuidae). Leaf resins of all Hymenaea species are comprised of essentially the same suite of sesquiterpene hydrocarbons, but discrete quantitative patterns have been classified as Compositional Types based on the amounts of major components. In this work pure extracts of leaf resins of Type I (high - and β-selinene), Type II (intermediate amounts of -and β-selinene and caryophyllene) and Type III (high caryophyllene) were incorporated into an artificial diet for the insects at 1 and 3.2% (dry wt). Resin Type treatments produced differential dose-dependent effects on growth rate (lower larval weights and increased time to pupation) and in mortality. More significant inhibitory effects occurred in Compositional Types with a predominance of a single compared (i.e. Types I and III). Significantly higher mortality occurring in Type III treatments suggests that caryophllene may have higher potential toxicity than - and β-selinene for S. exigua. These experiments indicate that feeding by generalist herbivores could be a factor determining quantitative compositional variation: (a) among populations; (b) during the development of leaves; (c) in the spatial distribution within the leaves; and (d) between parent tree and seedling progeny.     

RNA Polymerase B levels during the cell cycle ofPhysarum polycephalum

Summary Tritiated -amanitin has been used as a specific and sensitive probe to estimate the number of RNA polymerase B molecules in isolated nuclei, chromatin and nucleoids, obtained from macroplasmodia ofPhysarum polycephalum. During mitosis (metaphase±10 min) there is at least 10-fold less RNA polymerase B than at all phases of the cell cycle, even if DNA replication has been blocked in vivo. It is concluded that many of the RNA polymerase B molecules leave the chromatin during decondensation of the chromosomes in telophase of the synchronous nuclear division ofPhysarum.     

The maximum leaf surface temperatures of the higher plants observed in the Inland Sea area

The maximum leaf surface temperatures (MLSTs) of 126 species of higher plants were measured by means of an infrared thermometer, in the Inland Sea area, southwest of Honshu-Island, Japan, where plants suffered from severe environmental conditions due to an abnormally small amount of precipitation during the summer of 1978. The MLSTs of plants in the summer of 1978 were greater than or equal to those of 1979, when the environmental conditions were not so severe. The MLST measured in this study was 50.4 C for a non-succulent plant (Liriodendron), and 53.1 C for a succulent plant (Agave). Plants with different life forms appeared to have different MLSTs. The average of the MLSTs of conifers deciduous trees, and evergreens were 36.4, 37.7, and 40.3 C, respectively. This order corresponds to the distribution of forests from high to low, latitudes. Also the MLSTs were higher for woody plants than for herbaceous plants. Relatively high leaf temperatures were observed for climbing plants, both herbaceous and woody. Plants with narrow leaves had lower leaf surface temperatures than those with borad leaves. Herbaceous dicotyledonous plants actively growing at the end of the summer of 1978, in full sun at Hiroshima Castle were exclusively those with relatively high leaf temperatures.     

Relations between water content, potential and permeability in stems of conifers

Abstract. The influence of sapwood water content on the conductivity of sapwood to water was measured on stem sections of Pinus contorta. A reduction in relative water content from 100 to 90% caused permeability to fall to about 10% of the saturated value.
Pressure–volume curves of branchwood and stem sapwood of Pinus contorta and Picea sitchensis have been analysed to definè the tissue capacitance and the time constant and resistance for water movement between stored water and the functional xylem as functions of tissue water potential. Three phases in water loss were discernible. In the initial phase at high water potentials (> –0.5 MPa), the capacitance was large, the time constant long and the resistance to flow large in comparison with intermediate water potentials (−0.5 to −1.5 MPa). At still lower water potentials (−1.5 to −3.0 MPa), the time constant and resistance declined still further but the capacitance had a tendency to increase again, especially in the stemwood of Sitka spruce. Typical values in the second phase were for the time constant 5 s, for the resistance 4 × 10⁻¹³ N s m⁻⁵ and for the capacitance (change in relative water content per unit change in potential) 1×10⁻¹¹ m³ Pa⁻¹. These parameters define the availability of stored water and are being used in a dynamic model of water transport in trees.     

Study on the dehydrating effect of the red cell Na+/K+-pump in nystatin-treated cells with varying Na+ and water contents

Using the antibiotic Nystatin, we have developed a systematic method for the preparation of red blood cells with independently selected levels of intracellular Na⁺ concentrations and water content. Such cells provided an experimental model to study the effect of Na⁺/K⁺ pump stimulation on red cell water content. Even in initially dehydrated cells, stimulation of the Na⁺/K⁺ pump by elevated intracellular Na⁺ caused subsequent further loss of cell water. Cell water loss was reflected in decreased monovalent cation content per unit mass of hemoglobin and by a shift in the density distribution of the cell populations to higher densities on discontinuous Stractan gradients. We conclude that the 3 Na_out⁺ : 2 K_in⁺ stoichiometry of the Na⁺/K⁺ pump results in a net desalting effect with increased pump activity. Under the conditions of these experiments, the cell appears to have no effective mechanism to compensate for a net loss of ions and water.     

Extensive variation at the α-glycerophosphate dehydrogenase locus in species of waterstriders (Gerridae: Hemiptera)

Variation at the -glycerophosphate dehydrogenase (-Gpdh; EC 1.1.1.8) locus was surveyed in 11 species of waterstriders (Gerridae: Hemiptera) and five other species of aquatic Hemiptera. Species of waterstriders exhibited considerable inter- and intraspecific variation in degree of winglessness. Average heterozygosity (0.401±0.090) and average number of observed electromorphs (5.36±0.96) for the 11 gerrid species were well above values reported for nearly all other insect species surveyed to date. Wing-monomorphic and wing-polymorphic species did not differ in average -Gpdh heterozygosity. Of the three wing-polymorphic species surveyed geographically, two species exhibited marked variation in wing-morph frequencies but homogeneous -Gpdh allele frequencies. The third species exhibited geographically homogeneous -Gpdh and wing-morph frequencies, but no significant association between -Gpdh phenotype and wing morph was observed in any surveyed population. These results are consistent with hypotheses evoking either relaxed purifying selection at the -Gpdh locus in species of Gerridae due to the apparent reduced importance of flight, or selective maintenance of common -Gpdh electromorphs.This work was supported by NSF Grant DEB 76-20967 to Alan H. Brush, funds from the Research Foundation of the University of Connecticut to Carl W. Schaefer, and USPHS Grant GM 21133 to Richard K. Koehn.     

Mineral element content in Ulva lactuca L. with reference to eutrophication in Hong Kong coastal waters

The amounts of tissue nitrogen, phosphorus, potassium, sodium, calcium and iron were estimated in the green alga Ulva lactuca L. collected from 9 rural and 14 urban littoral sites in the waters around Hong Kong Island during 1978 and 1979. The mean levels of tissue nitrogen and phosphorus were respectively 65% and 87% more in urban sites than in rural ones. Very significant correlation (r = 0.920) was found between the logarithmic concentration of seawater inorganic nitrogen and that of tissue nitrogen. The same applied to soluble reactive phosphorus in seawater and tissue phosphorus (r = 0.886). The levels of potassium, sodium and calcium in the alga were relatively uniform. However considerable variation in the level of iron was detected. The potential use of Ulva as an indicator species for eutrophication is discussed.