An ultrasensitive colorimetric assay for manganese

An ultrasensitive colorimetric assay for manganese is described. It is based upon the catalysis, by Mn(II), of the photochemical oxidation of o-dianisidine, sensitized by riboflavin. Catalase increases the Mn(II)-catalyzed rate of photosensitized oxidation of dianisidine to the bisazobiphenyl, while superoxide dismutase inhibits the rate. The mechanism appears to involve oxidation of Mn(II) by O2-, followed by oxidation of dianisidine by MnO2+ in equilibrium Mn(III). Cu(II) interferes, but Zn(II), Fe(II), Fe(III), Co(II), and Ni(II) do not. Chelating agents and thiol reductants also interfere. Interference by Cu(II) can be overcome by the addition of cyanide, while interference by organic compounds can be surmounted by wet ashing. This assay provides a linear response to Mn(II) over the range 10-2500 nM. The limit of detection was 5 nM Mn(II).     

II. Selective accumulation of MPP+ in the substantia nigra: A key to neurotoxicity?

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is rapidly metabolized to a 1-methyl-4-phenylpyridinium species (MPP+) in the squirrel monkey. After administration of toxic doses of MPTP, the concentration of MPP+ in the substantia nigra appears to increase during the first 72 hours, reaching the highest concentration of any central nervous system (CNS) tissue studied. In contrast, the concentration of this compound in other brain areas suggested time dependent elimination during the same period. Pretreatment of animals with the monoamine oxidase (MAO) inhibitor pargyline blocks both the neurotoxic action and the biotransformation of MPTP. In animals given pargyline and MPTP, initial MPTP levels are much higher in all brain regions than in those not receiving pargyline, but by 12 hours, MPTP levels had fallen rapidly in all regions except the substantia nigra and the eye. It may be that the selective toxicity of MPTP is related in some way to the accumulation of its oxidized metabolite in the substantia nigra.     

Effects of ultraviolet-B radiation on plants during mild water stress. IV. The insensitivity of soybean internal water relations to ultraviolet-B radiation

The combined effects of ultraviolet-B (UV-B, 280–320 nm) radiation and water stress were investigated on the water relations of greenhouse grown soybean [ Glycine max (L.) Merr. cv. Essex]. On a weighted (Caldwell 1971), total daily dose basis, plants received either 0 or 3 000 effective J m² UV-B_BE supplied by filtered FS-40 sunlamps. The latter dose simulated the solar UV-B radiation anticipated at College Park, Maryland, U.S.A. (39°N latitude) in the event that the global stratospheric ozone column is reduced by 25%. Plants were either well-watered or preconditioned by drought stress cycles. Diurnal measurements of water potential and stomatal conductance were made on the youngest fully expanded leaf. Various internal water relations parameters were determined for detached leaves. Plants were monitored before, during and after water stress. There were no significant differences in leaf water potential or stomatal conductance between treatments before plants were preconditioned to water stress. However, drought stress resulted in significantly lower midday and afternoon leaf water potentials and lower leaf conductances as compared to well-watered plants. UV-B radiation had no additional effect on leaf water potential; however, UV did result in lower leaf conductances in plants preconditioned to water stress. Turgid weight:dry weight ratio, elastic modulus, bound water and relative water content were unaffected by UV-B radiation. Osmotic potentials at full and zero turgor were significantly lower in the drought stressed treatments as compared to well-watered plants.     

Identification of the coding region for the putidaredoxin reductase gene from the plasmid of Pseudomonas putida

The first 12 NH₂-terminal amino acids of the Pseudomonas putida putidaredoxin reductase were shown to be Met-Asn-Ala-Asn-Asp-Asn-Val-Val-Ile-Val-Gly-Thr. Comparison of these data with the DNA sequence of the BamHI-HindIII 197-base fragment derived from the PstI 2.2-kb fragment obtained from the P. putida plasmid showed that the putidaredoxin reductase gene was downstream from the cytochrome P-450 gene and the intergenic region had the 24-nucleotide sequence TAAACACATGGGAGTGCGTGCTAA. The Shine-Dalgarno sequence GGAG was detected in this region. The initiating triplet for the reductase gene was GTG, which normally codes for valine, but in the initiating codon position codes for methionine. From the amino acid sequence and X-ray data comparisons with other flavoproteins, what appears to be the AMP binding region of the FAD can be recognized in the NH₂-terminal portion of the reductase involving residues 5–35.     

Evidence for sterol carrier protein2-like activity in hepatic, adrenal and ovarian cytosol

Purified sterol carrier protein2 (SCP2) from rat liver stimulated utilization of endogenous cholesterol for pregnenolone synthesis by adrenal mitochondria. Cytosolic preparations of rat liver, adrenal and luteinized ovary were also stimulatory in mitochondrial pregnenolone synthesis to different extents. Treatment of all preparations with rabbit anti-rat SCP2 IgG neutralized the stimulatory effects, and immunoprecipitated proteins gave similar patterns on SDS-gradient polyacrylamide gel electrophoresis. Treatment with rabbit pre-immune IgG had no effect on these parameters. Thus, proteins which are immunochemically compatible with hepatic SCP2 appear to be present in steroidogenic tissues and may play a role in control of mitochondrial cholesterol side chain cleavage activity.     

Immunofluorescent localization of a murine seminal vesicle proteinase inhibitor

The indirect immunofluorescent technique was used to localize a proteinase inhibitor isolated from murine seminal vesicles. The inhibitor was found in the lumen and in the apical epithelium of the seminal vesicle but not in the testes, epididymides, ductus deferens or Cowper's glands. It was also associated with the anterior acrosomal region of ejaculated sperm and sperm recovered from the female tract within 5 min of coitus. The inhibitor is removed from uterine sperm between 2 to 4 h postcoitus, however sperm recovered from the uterus 2 h postcoitus will rebind inhibitor. The inhibitor is not normally associated with epididymal or ductus sperm although these sperm will bind purified inhibitor in vitro.     

The effects of time of seed production on the germination response of Spergularia marina

Germination studies were carried out with seeds of Spergularia marina L. Griseb produced over an interval of six months (June-November). The response of the seeds to light and dark, various constant and alternating temperature regimes, and salinity were determined. In addition, the effects of soil moisture status at the time of seed production on the subsequent germination response of seeds were also determined. Light was an absolute requirement for germination. While a constant temperature regime did not generally favour germination of seed of any month, alternating temperature greatly enhanced germination with an optimum at 5/15°C in all seeds. When imbibed in solutions of different salinities, seeds collected in July and October behaved like true halophyte seeds whereas those collected in June. August, September and November behaved like glycophyte seeds.
High concentration of gibberellic acid (3 000 μ M ) stimulated dark germination in the June and November seed lots, but in light, low GA3 concentration (300 μ M ) stimulated germination most. The addition of kinetin (30 μ M ) plus gibberellic acid enhanced germination in the dark in contrast to GA3 alone; kinetin alone stimulated a very low percentage germination.
The moisture status of the soil at the time seeds were produced did not affect the germination response of an early seed crop (July) but affected that of the later seeds (August).
Judging from the different germination responses, it appears that the seeds belong to at least two physiological groups, one which appears to need either a dark-wet or cold-wet pretreatmem for high germination to occur; and the other group which does not need pretreatmem. The ecological significance of these varied responses is discussed in relation to the survival of the species in its habitat.     

CAM-idling in Hoya carnosa (Asclepiadaceae)

In the leaf succulent Asclepiad Hoya carnosa (L.) R. Br., CAM photosynthesis occurred under well-watered conditions, as characterized by diurnal gas exchange and changes in titratable acidity. Following 10–12 days of severe water stress, the plants shifted from CAM to a modified CAM-idling mode of metabolism. CAM-idling was characterized by complete or almost complete stomatal closure accompanied by CAM-like diurnal changes in titratable acidity. H. carnosa plants maintained this CAM-idling mode of photosynthesis for at least 8 weeks. Upon reirrigation, the plants returned to the original CAM mode within 1 week. These results suggested that CAM-idling is a reversible, intermediate form of sustained metabolism which enables plant survival under conditions of extended drought.This work was supported in part by NSF Grant PCM 8200366 and in part by the Science and Education Administration of the United States Department of Agriculture under Competitive Grant 5901-0420-8-0018-0.     

Lectins from<Emphasis Type="Italic">Griffonia simplicifolia</Emphasis> seeds

The physical-chemical and carbohydrate binding specificity ofGriffonia simplicifolia I (GS I) isolectins, one of the 4 lectins isolated fromGriffonia simplicifolia seeds, are described.Association constants for the binding of methyl α- and β-D-galactopyranoside and methyl 2-acetamido-2-deoxy-α-D-galactopyranoside to the A₄, A₂ B₂ and B₄ isolectins are reported.Precipitation reactions of theGriffonia simplicifolia isolectins with guaran and type B blood group substance are described.The hypothesis that subunit B is a precursor of subunit A, a process involving proteolytic cleavage of the B subunit, was tested by conducting structural studies on the 2 subunits. The results indicated that the A and B subunits are probably products of 2 separate but closely related, possibly contiguous genes.     

Anisotropic Absorption of Linearly Polarized Light by Cylindrical Molecules

A general theory relating the orientation of the transition moment in a chromophore to incident light, linearly polarized at an arbitrary angle, is discussed for cylindrical molecules. Experimental verification of this theory is presented for molecules of tobacco mosaic virus (TMV), T₂ DNA, and polyadenylic acid (poly-A).