Sequestration,Phytoreduction, and Phytooxidation of Halogenated Organic Chemicals by Aquatic and Terrestrial Plants

Laboratory data from plant-mediated transformation of chlorinated and brominated alkanes, alkenes, and chlorinated pesticides, including phytotransformation data from field plants currently used in phytoremediation of trichloroethylene (TCE), were reviewed for the purpose of identifying important phytoprocesses and their respective roles in phytoremediation of halogenated organic compounds (HOCs). The results of the laboratory experiments indicated that the initial very rapid removal of hydrophobic HOCs from water or the gas phase by aquatic and terrestrial plants is primarily due to sequestration. The amount of HOC sequestered is controlled by the plant species and the physicochemical properties (e.g., K_ow, aqueous solubility, volatility) of the contaminant. Phytodegradation studies conducted in both the gas and aqueous phases indicated that hexachloroethane (HCA) is dechlorinated to the same metabolites by sterilized and axenically cultivated aquatic plants and an isolated plant dehalogenase factor. Similar results were obtained in experiments conducted with o,p'-DDT and p,p'-DDT in aqueous solution. The sterilized and axenically cultivated aquatic plants also oxidized HCA to similar chloroacetic acids. The metabolism of HOCs to the corresponding oxidative and reductive transformation products identified in the plant rhizosphere, stems, and leaves suggested that more than one pathway, requiring different enzymes, may be involved in phytotransformation reactions. Four phytoprocesses (mechanisms) were found to be important in the removal of the probe HOCs from water by aquatic plants, namely, (1) rapid sequestration by partitioning to the lipophilic plant cuticles; (2) phytoreduction to less halogenated metabolites; (3) phytooxidation to haloethanols, haloacetic acids, and unidentified metabolites; and (4) assimilation into the plant tissues as nonphytotoxic products, presumably produced by covalent binding with the plant tissues. Laboratory and field data indicate that the distribution of metabolites of perchloroethylene (PCE) and TCE in cottonwood and willow trees is determined by the growth stage or age of these vascular plants, the plant species, and the duration of exposure to the compound. For terrestrial plants, the predominant phytoprocesses by which HOCs are attenuated in the environment include sequestration, rhizodegradation, uptake, phytodegradation, and phytovolatilization. Using PCE as a model chlorinated organic solvent, possible phytotransformation pathways are proposed to account for the different metabolites identified in the rhizosphere and tissues of laboratory and field plants. The proposed pathways also combine phytoreduction reactions that occur in plant tissues and are likely catalyzed by plant dehalogenase(s) for example, enzyme(s) such as glutathione-S-transferase and Fe-S clusters in chloroplast ferredoxin, with phytooxidation and covalent binding (phytoassimilation) reactions mediated by oxidative-enzymes (possibly cytochrome P-450 with monooxygenase activity, glutathione or laccase). Depending on the characteristics of the field site, the phytoprocesses identified in this study are vital in the design and implementation of phytoremediation of halogenated organic contaminants.     

Metabolic Effects of Mental Stress during Over- and Underfeeding in Healthy Women

Objective: To assess the short-term consequences of carbohydrate or fat overfeeding or of food restriction on the metabolic effects of mental stress in healthy lean women. Research Methods and Procedures: The effects of a sympathetic activation elicited by mental stress were evaluated in a group of healthy women after standardized isocaloric feeding (ISO) or after a 3-day overfeeding with 40% excess calories as either carbohydrate overfeeding (CHO OF) or fat overfeeding (FAT OF). Oxygen consumption rate (VO ₂) was measured as an index of energy expenditure, and subcutaneous glycerol concentrations were monitored with microdialysis. The same measurements were performed in another group of healthy women after ISO and after a 3-day period of underfeeding with a protein sparing modified fast (UF). Results: In all conditions, mental stress significantly increased heart rate, blood pressure, plasma norepinephrine and epinephrine concentrations, and VO ₂, and produced a nonsignificant increase in subcutaneous glycerol concentrations. CHO OF and FAT OF did not alter the effects of mental stress on VO ₂ and subcutaneous glycerol concentrations. In contrast, UF increased basal VO ₂ but significantly reduced its stimulation by mental stress. UF also enhanced the increase in subcutaneous glycerol concentrations during mental stress. Discussion: UF reduces the stimulation of energy expenditure and enhances lipolysis during sympathetic activation. These adaptations may be involved in mobilization of endogenous fat while limiting weight loss. In contrast, short-term overfeeding fails to alter the sympathetic control of energy expenditure and lipolysis.     

In vivo peroxidative activity of FALS-mutant human CuZnSODs expressed in yeast.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder leading to loss of motor neurons. We previously characterized the enhanced peroxidative activity of the human familial ALS (FALS) mutants of copper-zinc superoxide dismutase (CuZnSOD) A4V and G93A in vitro. Here, a similar activity is demonstrated for human FALS CuZnSOD mutants in an in vivo model system, the yeast Saccharomyces cerevisiae. Spin trap adducts of alpha-(pyridyl-4-N-oxide)-N-tert-butylnitrone (POBN) have been measured by electron paramagnetic resonance (EPR) in yeast expressing mutant (A4V, L38V, G93A, and G93C) and wild type CuZnSOD upon addition of hydrogen peroxide to the culture. The trapped radical is a hydroxyethyl adduct of POBN, identified by spectral parameters. Mutant CuZnSODs produced greater concentrations of the trapped adduct compared to the wild type enzyme. This observation provides evidence for an oxidative radical mechanism, whereby the mutants of CuZnSOD catalyze the formation of reactive oxygen species that may be related to the development or progression of FALS. This study also presents an in vivo model system to study free radical production in FALS-associated CuZnSOD mutations.     

One-kilobase direct repeats of plasmid pSa

One-kilobase, direct repeats were found on either side of the chloramphenicol resistance gene of plasmid pSa. The right repeat corresponded to the region coding for sulfanilamide resistance. The repeats were not identical as judged by distances between restriction enzyme sites, hybridization, and by the ability to confer resistance to sulfanilamide.     

Purification and characterization of cardiac tropomyosins.

A new procedure was developed to purify tropomyosin. The procedure was an adaptation of that described for purification of myosin. By eliminating troponin before precipitating with (NH4)2 SO4, it was possible to obtain pure tropomyosin from the same preparation from which myosin was purified. When tropomyosin was subjected to isoelectrofocusing two tropomyosins were present, having similar isoelectric points of pH 5.4 and 5.6; two tropomyosin subunits were resolved in the presence of 6 M urea. The two subunits had very similar isoelectric points, pH 4.7 and 5.0. According to Ouchterlony analyses the tropomyosins from canine skeletal and cardiac tissue were immunologically identical when incubated with goat gammaG antitropomyosin (cardiac).     

Dynamics of fd coat protein in the bacteriophage

The dynamics of the coat protein in fd bacteriophage are described with solid-state 15N and 2H NMR experiments. The virus particles and the coat protein subunits are immobile on the time scales of the 15N chemical shift anisotropy (10(3) Hz) and 2H quadrupole (10(6) Hz) interactions. Previously we have shown that the Trp-26 side chain is immobile, that the two Tyr and three Phe side chains undergo only rapid twofold jump motions about their C beta-C gamma bond axis [Gall, C. M., Cross, T. A., DiVerdi, J. A., & Opella, S. J. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 101-105], and that most of the backbone peptide linkages are highly constrained but do undergo rapid small amplitude motions [Cross, T. A., & Opella, S. J. (1982) J. Mol. Biol. 159, 543-549] in the coat protein subunits in the virus particles. In this paper, we demonstrate that the four N-terminal residues of the coat protein subunits are highly mobile, since both backbone and side-chain sites of these residues undergo large amplitude motions that are rapid on the time scales of the solid-state NMR experiments. In addition, the dynamics of the methyl-containing aliphatic residues Ala, Leu, Val, Thr, and Met are analyzed. Large amplitude jump motions are observed in nearly all of these side chains even though, with the exception of the N-terminal residue Ala-1, their backbone peptide linkages are highly constrained. The established information about the dynamics of the structural form of fd coat protein in the virus particle is summarized qualitatively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Benzodiazepine receptor mediated discriminative cues: Effects of GABA-ergic drugs and inverse agonists

Rats were exposed to a two-layer drug discrimination procedure using the benzodiazepine (BZ) receptor inverse agonists N′-methyl-β-carboline-3-carboxamide (FG 7142) or methyl-6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM). FG 7142 (30 mg/kg) failed to acquire discriminative stimulus control, although it did suppress responding. The same group of animals was trained successfully to discriminate diazepam (DZP, 2.5 mg/kg) from vehicle. The DZP cue was potentiated by the GABA agonist 4,5,6,7-tetrahydro-isoxazolo [5, 4-c] pyridin-3-ol (THIP, 1–3 mg/kg); THIP alone produced vehicle-appropriate responding. In addition, clonazepam (0.2 mg/kg) and chlordiazepoxide (5 mg/kg) substituted for DZP (with potencies of 7.5 and 0.25 times that of DZP, respectively). In antagonism tests, FG 7142 (5–17.5 mg/kg), methyl-β-carboline-3-carboxylate (β-CCM, 2.5 mg/kg), nicotine (0.3 mg/kg), harmaline (5 mg/kg) and naltrexone (10 mg/kg) did not effect, bicuculine (2 mg/kg) and DMCM (1 mg/kg) partially blocked, and the BZ receptor antagonist Ro 15–1788 (40 mg/kg) completely blocked the discriminative stimulus effects of DZP. In animals trained to discriminate DMCM (0.2 mg/kg) from vehicle, 95% substitution occured with bicuculline (2 mg/kg); DZP (1–5 mg/kg) completely antagonized DMCM. These results indicate that the DZP cue is mediated by GABA-coupled BZ receptors and that GABA may modulate the efficacy of a BZ at its receptor site. However, since inverse BZ receptor agonists (FG 7142, DMCM and β-CCM) were, at best, only marginally effective in antagonizing DZP, the DZP cue may be mediated by a distinct subclass of BZ receptors.