Distribution of gibberellin-like substances in light-and dark-grown seedlings of Phaseolus multiflorus

Summary Lettuce and barley endosperm bioassays of successive eluates from a phosphate-buffered celite column detected two gibberellin-like compunds, Phaseolus I and Phaseolus II, in extracts of both light-and dark-grown seedlings of Phaseolus multiforus. There were differences in the gibberellin content of light-and dark-grown seedlings, the former containing more Phaseolus I but less Phaseolus II than the latter. An examination of the gibberellin content of leaves and apical buds, stems, cotyledons, and roots of light-and dark-grown seedlings revealed distinct qualitative and quantitative differences in the distribution of Phaseolus I and Phaseolus II.     

Sulfation of hypertensive and hypotensive drugs by monkey brain phenol sulfotransferase

The substrate specificity and affinity of two forms of phenol sulfotransferase (PST) from Rhesus macaque brain cortex were studied. Catecholamines, their methylated metabolites (normetanephrine, metanephrine) and methylated precursor, -methylDOPA, were examined as substrates for both the cationic (PST I) and the anionic (PST II) forms of the enzyme. Sulfation of hypertensive drugs (phenylephrine, octopamine, metaraminol), hypotensive drugs (-methylDOPA, minoxidil), and related agents without a free hydroxy group on the benzene ring were also studied. Results indicated that both PST forms sulfated -methylDOPA and minoxidil, but only PST II transferred the sulfate group to catecholamines and most of the adrenergic agents examined.     

Biochemical characteristics of primary and passaged cultures of primate brain microvessel endothelial cells

Rhesus macaque monkey brain microvessel endothelial cells (BMECs) were isolated and grown in culture in an effort to establish an appropriate primate in vitro model of the endothelial component of the blood-brain barrier. The presence of Factor VIII antigen, alkaline phosphatase, -glutamyl transpeptidase, lactate dehydrogenanse, total protein, and the passive permeability properties was documented for both primary and passaged cultures. Primate BMECs were shown to exhibit similar morphological and biochemical properties described for other BMEC culture systems derived from other species. In addition, the passaged primate BMECs were particularly notable for the changes in enzyme activities and total protein that parallel age-dependent changes in brain capillary endothelia. This study provides further support for the possible application of BMEC culture systems in investigations of blood-brain barrier functions under normal, aging, and diseased conditions.To whom to address reprint requests. Phone (913)864-3609; FAX (913)864-3578.     

Gestational and smoking effects on peptidase activity in the placenta

Angiotensin converting enzyme (ACE) and leucine aminopeptidase (LAP) regulate fetally and maternally generated peptides in the placenta. In this study, ACE-like activity was found to be decreased and LAP-like activity increased with increasing days of gestation in rat placental tissues forming the fetal:maternal interface. Membrane-associated ACE-like and LAP-like activities in the placenta of smokers were also found to be significantly higher than their respective activities in placenta of nonsmokers. Our collective findings suggest that gestational and environmentally-induced changes in placental peptidase activities may account for variable peptide hormone and/or therapeutic peptide metabolism in the placenta.     

Catecholamine-Metabolizing Enzymes of Bovine Brain Microvessel Endothelial Cell Monolayers

The activities of monoamine oxidase (MAO), catechol-O-methyltransferase (COMT), phenol sulfotransferase (PST), alkaline phosphatase (AP), gamma-glutamyl transpeptidase (GT), and angiotensin converting enzyme (ACE) were quantitated in primary cultures of bovine brain microvessel endothelial cell monolayers and cerebral gray matter. Significant MAO-A and -B, cytosolic and membrane-bound COMT, PST, AP, GT, and ACE activities are demonstrated in bovine gray matter. By comparison, enzyme activities of the monolayers vary with the age of the monolayer and are generally higher in complete monolayers. Relative to gray matter enzyme activities, the monolayers are enriched with AP, GT, and ACE, enzymes considered to be markers for brain endothelium. Results also indicate that the activities of MAO-A and PST in the monolayers approach those found in the gray matter. Conversely, cytosolic COMT and MAO-B activities in the monolayers are negligible and much lower, respectively, compared to activities in gray matter. Additional studies with both tissues suggest that the PST of both tissues is the thermostable form of the enzyme.     

The biological detoxication of hormone herbicides in soil

Summary (1) The results of experiments on the continuous perfusion of aerated solutions of the herbicides 2,4-dichlorophenoxyacetic acid (2,4-D), 2, Methyl-4, chlorophenoxyacetic acid (MCPA) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) through garden soil indicate that the kinetics of their breakdown are essentially similar.(2) Three phases can be distinguised: (a) an immediate initial adsorption onto soil colloids amounting to 0.167±0.035 mg per g dry soil, (b) a lag phase of varying duration in which there is little or no disappearance of herbicide, (c) a final phase of rapid complete detoxication. Such enriched soils detoxicate subsequent perfusions with the same herbicide molecule at a constant high rate. Enrichment times for 2,4-D, MCPA and 2,4,5-T are roughly of the order or 14, 70 and 270 days respectively.(3) This indirect evidence, from kinetics of detoxication, of the development of a bacterial flora adapted to herbicide breakdown, is strongly supported by experiments in which the detoxicating activity of such enriched soils was completely destroyed by low concentrations (0.01%) of the bacterial poison, sodium azide.(4) Attempts to isolate in pure culture the 2,4-D-decomposing organism were successful. Preliminary classification work relegates the organism to the Bacterium globiforme group. Further confirmatory work is required.(5) Cross perfusion experiments in which 2,4-D enriched soils were perfused with MCPA and vice versa indicate that both can detoxicate the other analogue, although at a somewhat lower rate than the soil directly enriched with that analogue.(6) 2,4-D enriched soil will not detoxicate 2,4,5-T although some activity may be temporarily induced by an intermediate perfusion with MCPA.