Proteomic analysis of methyl parathion-responsive proteins in zebrafish (Danio rerio) brain

Methyl parathion (MP), an organophosphorus pesticide used worldwide, has been associated with a wide spectrum of toxic effects on organisms in the environment. This study set out to analyze the alteration of protein profiles in MP-exposed zebrafish (Danio rerio) brain and find the proteins responsive to MP toxicity. Zebrafish were subjected to 1, 3 and 5 mg/L MP and the proteomic changes in their brains were revealed using two-dimensional gel electrophoresis. Six protein spots were observed to be significantly changed by MP exposure. Among these, 4 spots were down-regulated, while 2 spots were up-regulated. These altered spots were excised from the gels and identified by matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry and database searching. The results indicate that these proteins were involved in binding, catalysis, regulation of energy metabolism and cell structure. These data may provide novel biomarkers for the evaluation of MP contamination and useful insights for understanding the mechanisms of MP toxicity.     

Updated Risk/Benefit Analysis of Fish Consumption Effects on Neurodevelopment: Implications for Setting Advisories

Epidemiological studies indicate that fish consumption during pregnancy leads to improved neurodevelopment. This suggests that the beneficial nutrients in fish may offset the adverse effects of mercury in the case of the average fish meal. However, our previous risk/benefit model predicted a net neurodevelopmental risk for the majority of species analyzed. In this article the previous model is calibrated against fish benefit data and then compared to other fish risk/benefit models, including recent models from the World Health Organization and USFDA. Our calibrated model estimated greater benefit for low mercury species but greater risk for high mercury species than the other models. With respect to a commonly eaten high mercury fish, swordfish, the calibrated model yielded risks that are supportive of current fish advisories but, in contrast, the other models predicted net neurodevelopmental benefits. The calibrated model was used in a proposed 3 step framework for setting fish consumption advisories: (1) Set initial consumption level based upon mercury RfD; (2) Adjust consumption upward if risk/benefit model indicates a net benefit; (3) Cap fish consumption based upon saturation of O-3 benefit. The implications of this approach for 7 varieties of fish are used to illustrate the framework.     

Model studies on methyl amyloses: correlation between reaction conditions and primary structure

Methyl amyloses have been prepared under various conditions and studied as model compounds for the determination of the substitution pattern in the polymer chain. After permethylation with iodomethane-d₃ the glucosidic linkages were statistically cleaved by partial methanolysis or reductive cleavage. The distribution of substituents in the dimer-, trimer-, and tetramer fraction was determined by FAB-MS and MALDI-TOF-MS and compared with that calculated from the monomer composition. While a homogeneous methylation in water gave the expected random distribution, a reaction in Me₂SO solution with sodium hydroxide and iodomethane yielded a methyl amylose with a surprising bimodal substitution pattern. A third example indicates a ds gradient in the sample as a result of topochemical reaction control.     

Chemistry of methylcorrinoids related to their roles in bacterial C1 metabolism

Abstract Corrinoids are central cofactors in bacterial metabolism, where they participate in a series of organometallic and redoxprocesses. These depend on the unique coordination chemistry and reactivity of the corrin-bound cobalt centers to which, in the complete corrins, also a nucleotide function can coordinate intramolecularly. The roles of methylcorrinoids in bacterial C₁ metabolism focus around the unusual Co-C-bond.     

Isolation and characterization of the carbohydrate moiety bound to N-7-mercaptoheptanoyl-O-threonine phosphate

Abstract Component B ( N -7-mercaptoheptanoyl-threonine- O -3-phosphate) (HS-HTP) which is an absolute requirement in the methylcoenzyme M methylreductase reaction was found to be part of a complex UDP-disaccharide when isolated carefully from cell-free supernant of Methanobacterium thermoautotrophicum . The site of attachment of HS-HTP to the UDP-disaccharide was through a carboxylic-phosphoric anhydride linkage of the C-6 mannosaminuronic acid to the phosphate group in HS-HTP. This bond is quite labile and this may account for the fact that the intact molecule, called methyl reducing factor (MRF) was not isolated previously. The structure of MRF was determined by combined fast atom bombardment mass spectrometry and ¹H-, ¹³C-, and ³¹P-NMR spectroscopy and assigned as: uridine 5'-[ N -7-mercaptoheptanoyl- O -3-phosphothreonine(2-acetamido-2-deoxy- β -mannopyranuronosyl)acid anhydride]-(1 → 4)- O -2-acetamido-2-deoxy α -glucopyranosyl diphosphate.     

β-Mannanolytic system of Aureobasidium pullulans

A xylanolytic yeast strain Aureobasidium pullulans NRRL Y 2311-1, was found to produce all enzymes required for complete degradation of galactomannan and galactoglucomannan. The enzymes differed in function and cellular localization: endo-β-1,4-mannanase was secreted into the culture fluid, β-mannosidase was strictly intracellular, and α-galactosidase and β-glucosidase were found both extracellularly and intracellularly. Among these enzyme components, only extracellular β-mannanase and intracellular β-mannosidase were inducible. The production of β-mannanase and β-mannosidase was 10- to 100-fold higher in galactomannan medium than in medium with one of the other carbon sources. β-mannanase and β-mannosidase were coinduced in glucose-grown cells by galactomannan, galactoglucomannan, and β-1,4-manno-oligosaccharides. The natural inducer of extracellular β-mannanase and intracellular β-mannosidase appeared to be β-1,4-mannobiose. Synthesis of both enzymes was completely repressed by glucose, mannose, or galactose. The synthetic glycoside methyl β-d-mannopyranoside served as a nonmetabolizable inducer of both β-mannosidase and β-mannanase. Received: 24 June 1996 / Accepted: 26 September 1996     

Cost-effectiveness of five burrow fumigants for managing black-tailed prairie dogs

We evaluated the cost-effectiveness of two solid form burrow fumigants (aluminum phosphide and gas cartridges) and three pressurized gas–liquid burrow fumigants (methyl bromide, chloropicrin, and a methyl bromide–chloropicrin mixture) for managing black-tailed prairie dogs (Cynomys ludovicianus). Fifty-two variable-sized plots, including 25 treatment and 25 control burrows, were established within 13 prairie dog colonies in central Nebraska during spring 1989. Each group of 25 treatment burrows was fumigated with one of the five fumigants according to label directions or manufacturer recommendations. All five fumigants reduced burrow activity 95–98%, as measured by a plugged burrow technique. No significant differences in efficacy (P=0.453) were detected among the five treatments. Total costs for materials and labor for the aluminum phosphide and gas cartridges, excluding application equipment, were twice ($75.00 to $96.88 ha⁻¹) the cost of the pressurized gas–liquid fumigants ($37.67 to $41.76 ha⁻¹). Costs for the application equipment were considerably higher for the pressurized materials. Each treatment required labor for burrow plugging, which accounted for 50–75% of the total cost. None of the products tested met all requirements of a proposed selection criteria for fumigants.     

Alleviation of drought stress and the physiological mechanisms in Citrus cultivar (Huangguogan) treated with methyl jasmonate

ABSTRACT

The role of exogenous methyl jasmonate (MeJA) in alleviating drought stress was investigated on Huangguogan. Except for intercellular CO₂ concentration, MeJA had little effect on net photosynthetic rate, stomatal conductance, and transpiration rate under drought stress. Compared with drought stress, MeJA significantly alleviated the decrease of chlorophyll content. However, chlorophyll a/b ratio was significantly increased. MeJA significantly increased proline and soluble sugar contents, significantly decreased the O₂ ^?· and H₂O₂ levels, and increased SOD and POD activities. In addition, the MDA content of drought stress was the highest of all treatments. MeJA significantly reduced MDA content in drought-stressed Huangguogan leaves. Although the Ascorbic acid (AsA) contents of 500 and 1000 mg L^?1 MeJA treatments were lower than that of 250 mg L^?1 MeJA, but all concentration of MeJA treatments delayed the decline of AsA content. Therefore, MeJA could induce drought stress tolerance by increasing the osmotic adjustment substances and antioxidant activities.     

500 MHz 1H-NMR study of the N-terminal N-trimethylalanine residue of LC-1 and LC-2 light chains m rabbit fast skeletal myosin solutions

High-resolution proton NMR at 500MHz has been used to study the N-trimethyl terminals of LC-1 and LC-2 light chains in rabbit fast skeletal myosin solutions. The observed resonance is a sum of two Lorentzians withΔv = 5 ± 1 Hz, δ = 3.23 ppm and Δv = 12 ± l Hz, δ = 3.22ppm, respectively. By selective proteolytic modifications samples lacking either the N-terminal segment of LC-1 up to Lys-17 (papain modification) or the N-terminal segment of LC-2 up to Arg-7 (trypsin modification) were prepared. From the NMR spectra of the modified samples the narrower -N⁺(CH₃)₃ methyl resonance is shown to originate from LC-1 and the broader from LC-2. Thus in solution LC-1 and LC-2 N-terminals do not behave identically and there exists between both terminals no interaction reflected by linewidth or chemical shift variation when either N-terminal is removed. In intact as well as in modified myosins the number of resonating protons corresponds within experimental error to the expected values. The comparison of the present NMR results with the rates of proteolytic cleavage suggests that in solution these light chains could be folded back along S1 but without impeding the motion of the N-terminal residues, while in filaments the LC-1 and LC-2 light chains would expose their median sensitive part to proteolytic enzymes.