Multienzyme inhibition assay for residue analysis of insecticidal organophosphates and carbamates

A recently developed spectrophotometric assay for the detection of organophosphorus and carbamate insecticides by means of cutinase inhibition has been successfully extended to two esterases derived from Bacillus subtilis (BS2) and rabbit liver. These esterases were selected because of their high sensitivity to the examined insecticide classes and their pronounced inhibition profile. With inhibition constants (ki) of 2.0x10(7) and 2.6x10(6) L/(mol.min) for rabbit liver esterase and BS2, respectively, chlorpyrifos oxon proved to be the strongest inhibitor directly followed by paraoxon. As compared to choline esterases and the recently studied cutinase, both esterases are surprisingly strongly inhibited by organophosphorus thions, showing k i in the range of 5.3x10(2) to 2.3x10(4) L/(mol.min). All tested insecticidal carbamates were also inhibitors of BS2 and rabbit liver esterase, albeit in a rather uniform manner. Generally, both enzymes were found to be about 2 orders of magnitude more sensitive on the studied insecticides than cutinase even with an enhanced sensitivity against plant matrix effects. Plant extracts, obtained according to the QuEChERS method, were subjected to solid-phase extraction (SPE) using a mixed mode strong anion exchanger/primary secondary amine sorbent and C18endcapped cartridges for superior cleanup. With spiked samples of apple juice, best recoveries of 73% (+/-61%), 94% (+/-25%), and 134% (+/-17%) were obtained for chlorpyrifos, parathion-methyl, and paraoxon, respectively. Results of exemplarily performed liquid chromatography-mass spectrometry control measurements were well in accordance with measurements obtained by enzyme inhibition.     

Synthesis and insecticidal activities of novel N-sulfenyl-N'-tert-butyl-N,N'-diacylhydrazines. 3. N-(alkyldithio), n-(aminothio), and N,N-dithio derivatives

A series of novel N-(alkyldithio) and N-aminothio derivatives of N'-tert-butyl-N,N'-diacylhydrazines were designed and synthesized as insect growth regulators via the key intermediates N-chlorosulfenyl-N'-tert-butyl-N,N'-diacylhydrazines. A series of novel N,N-dithio derivatives were also designed and synthesized with N'-tert-butyl-N,N'-diacylhydrazines and sulfur chloride in the presence of sodium hydride. Compared to the parent compounds, these derivatives displayed better solubility and improved hydrophobicities. The insecticidal activities of the target compounds were evaluated. The results of bioassays showed that the target compounds possessed comparable activities against oriental armyworm with the corresponding parent compounds. In particular, N-(morpholinosulfenyl)-N'-tert-butyl-N-4-ethylbenzoyl-N'-3,5-dimethylbenzoylhydrazide (IIIk) has higher toxicities against oriental armyworm and diamondback moth than the parent compound, RH-5992.     

Oxidation of ascorbate in raw milk induced by enzymes and transition metals.

The effect of xanthine oxidase, lactoperoxidase, and transition metals [Fe(III), Cu(II)] on the oxidation of ascorbate in raw milk was investigated. Data clearly showed that iron(III) (200 microM) does not accelerate ascorbate oxidation in raw milk in concentrations relevant for raw milk. In contrast, addition of copper(II) (10 microM) to the raw milk accelerated oxidation of ascorbate. Furthermore, both xanthine oxidase and peroxidase activity were found to accelerate ascorbate oxidation dramatically in raw milk, indicating that xanthine oxidase and lactoperoxidase might be some of the most obvious candidates for mediation of ascorbate oxidation in raw milk. The present data are discussed in relation to using the fate of ascorbate in raw milk as an indicator of the oxidative stability of the milk.     

Removal of Fatty Acids from Palm Oil Effluent by Combined Electro-Fenton and Biological Oxidation Process

The objective of this paper is to design a pilot plant electrochemical reactor and to prove the operational concept of the electrochemical production of ferrate in situ and its online application for sewage treatment. To that end, the first part of this paper focuses on the analysis of the main engineering aspects of the reactor and the electrochemical process that affect the ferrate production, using laboratory scale experiments such as the interelectrode gap, the space-time yield, the area/volume (A/V) ratio, the current efficiency, and the energy consumption. The second part focuses on the production of ferrate using a pilot plant scale to prove the operational concept of the electrochemical generation of ferrate in situ and its online application as a step towards its full scale application for water and wastewater treatment.     

Two inhibition targets by [Cr(2gb)(3)](3+) and [Co(2gb)(3)](3+) on redox enzymes of spinach thylakoids.

[Cr(2gb)(3)]Cl(ZnCl(4)), [Cr(2gb)(3)]Cl(3), and [Co(2gb)(3)]Cl(3) were synthesized and characterized. Their chemical structures and the oxidation states of their metal centers remained unchanged in solution. The effects of these compounds, CrCl(3) and [Co(NH(3))(6)]Cl(3), on photosynthesis were investigated. The coordination compounds inhibit ATP synthesis and electron flow (basal, phosphorylating, and uncoupled), behaving as Hill reaction inhibitors. The target for [Cr(2gb)(3)]Cl(ZnCl(4)) is located at the Q(B) level. In contrast, the interaction sites of [Cr(2gb)(3)]Cl(3) and [Co(2gb)(3)]Cl(3) are located in the span from P(680) to Q(A) and at the b(6)f complex. Neither CrCl(3) nor [Co(NH(3))(6)]Cl(3) inhibited photosynthesis. The 100% inhibition on PS II of [Cr(2gb)(3)]Cl(ZnCl(4)) is explained in terms of a synergystic effect between the 2gb-chromium(III) coordination compound and the ZnCl(4)(2)(-) anion.     

Organophosphate inhibition of an arylesterase from Lumbricus terrestris and reversal by pralidoxime

Several enzymes of xenobiotic metabolism have been identified in soil annelids. Their roles in the physiology and ecotoxicology of the organism are diverse and may parallel those found in mammalian species. Among these are the arylesterases. An arylesterase assayed by hydrolysis of p-nitrophenylacetate was identified in Lumbricus terrestris. This arylesterase appears to be a serine esterase, since it is sensitive to inhibition by metrifonate. This mechanism of inhibition is similar to that of cholinesterase, since pralidoxime can reverse the inhibition of arylesterase by metrifonate. The presence of this esterase in soil annelids may provide insight into the processing of environmental chemicals by soil annelids as well as in vitro commercial applications of esterases based upon their mechanism of catalysis and inhibition.     

超声波辅助制备米糠生物柴油及其燃料排放特性

对超声波辅助碱催化米糠油与醇酯交换反应进行了一些列试验研究,并与传统机械搅拌法比较,论证其用于制备生物柴油的可行性.结果显示超声波辅助方法可以缩短酯交换反应的时间15～20 min.超声频率40 kHz条件下反应速度较28 kHz快,但是由于产物水洗分离困难而使得产物得率反而有所下降.超声辅助酯交换法还可以减少催化剂的用量,当催化剂用量为质量分数0.5%时可得到最高的转化率和得率.轻型车排放试验结果显示燃用米糠生物柴油可以显著降低CO和HC排放,而Nox排放有少量增加.     

Rapid and complete extraction of phenols from olive oil and determination by means of a coulometric electrode array system

Extraction methods to determine olive oil phenols are not exhaustive. A procedure to test their effectiveness, based on the treatment of the extracted oil with 2 N HCl followed by analysis of phenols in the aqueous phase, has been developed. It was concluded, using this test, that 15-40% of phenols remained unextracted when the liquid/liquid extraction method with 80% methanol was applied. Solid phase extraction (C(18) cartridge) succeeded in retaining most of the phenols in the cartridge, but the recovery yield from the sorbent material was low. However, a new extraction method, based on the use of N,N-dimethylformamide (DMF) as an extraction solvent, achieved a complete extraction of phenols from oils. The proposed method requires a lower amount of oil, solvents, energy, and labor than the traditional ones. Because of the low concentration of phenols in the DMF extract, the highly sensitive electrochemical detector (EC) technique was studied. All of the phenols detected by the traditional UV detectors were also detected by EC using a coulometric array system. A rapid and complete analytical methodology of phenols in olive oil has been proposed based on coupling DMF extraction and EC detection.     

Abundance of Iron-Oxidizing Thiobacilli and Biological Sulfur Oxidation Potential from Soil Impacted by Coal and Coal Refuse Piles

This study was conducted to assess the abundance of iron-oxidizing bacteria and biological sulfur oxidation potential from soil impacted by coal and coal refuse from two coal-burning electric power facilities located at the U.S. Department of Energy's Savannah River Site (Aiken, S.C.) and the South Carolina Electric and Gas Site at Beech Island, S. C. Significantly higher MPN counts of iron-oxidizing bacteria were obtained from samples collected at the confluence of a coal storage runoff containment basin, a coal reject (refuse) pile, and an adjacent wetland at the Savannah River Site. Significant differences in pH, sulfate-S, ferrous- and ferric-iron were also obtained between sampling locations. No significant differences in ferric/ferrous ratios were determined. These ratios however, exceeded a value of 2.0 when sample pH values were less than 4.5. Under optimal conditions, biological thiosulfate-S oxidation potentials (in vitro) showed a 4- to 7-day lag in the appearance of sulfate-S, and a final pH (after twenty-four days of perfusion) of 1.97 to 3.90. These results indicate that contamination of subsurface water by acidic leachate derived from thionic bacterial activity will occur if coal and coal refuse piles are not confined by an impermeable surface or containment facility.     

Antioxidant levels and inhibition of cancer cell proliferation in vitro by extracts from organically and conventionally cultivated strawberries

The effects of extracts from five cultivars of strawberries on the proliferation of colon cancer cells HT29 and breast cancer cells MCF-7 were investigated, and possible correlations with the levels of several antioxidants were analyzed. In addition, the effects of organic cultivation compared to conventional cultivation on the content of antioxidants in the strawberries and strawberry extracts on the cancer cell proliferation were investigated. The ratio of ascorbate to dehydroascorbate was significantly higher in the organically cultivated strawberries. The strawberry extracts decreased the proliferation of both HT29 cells and MCF-7 cells in a dose-dependent way. The inhibitory effect for the highest concentration of the extracts was in the range of 41-63% (average 53%) inhibition compared to controls for the HT29 cells and 26-56% (average 43%) for MCF-7 cells. The extracts from organically grown strawberries had a higher antiproliferative activity for both cell types at the highest concentration than the conventionally grown, and this might indicate a higher content of secondary metabolites with anticarcinogenic properties in the organically grown strawberries. For HT29 cells, there was a negative correlation at the highest extract concentration between the content of ascorbate or vitamin C and cancer cell proliferation, whereas for MCF-7 cells, a high ratio of ascorbate to dehydroascorbate correlated with a higher inhibition of cell proliferation at the second highest concentration. The significance of the effect of ascorbate on cancer cell proliferation might lie in a synergistic action with other compounds.     

Specific and non-specific adsorption of inorganic ions I. Evaluation of specific adsorbability by means of minimum concentration for specific adsorption

The Minimum Concentration for Specific Adsorption (MCSA) was defined as a reversal of the charge concentration of the ion at the infinite dilution of sol (adsorbent), or practically at such concentration of sol that equilibrium of the ion Is but negligibly affected by adsorption of the ion.

The MCSAs of 15 species of cations were measured with respect to SiO₂ at pH 4,6,5, and 10; the MCSAs of 10 species of anions were measured with respect to Fe(OH)₃ at pH 4, and 6.5; and the MCSAs of 10 species of anions were measured with respect to Al-coated-SiO₂ at pH 6.5. Relative specificities of the inorganic ions based on the MCSAs at pH 6.5 were as follows:

The MCSAs of cations with respect to SiO₂ and of anions to Fe(OH)₃ and Al-coated-SiO₂ at pH 6.5 had close relationships to ionic potentials and general solubility of the elements in water. Based on these results, the authors concluded that the MCSA could be used u an index of specificity (i.e. covalent bond-forming tendency) of an Jon at adsorption reactions.

The values corresponding to the apparent stability constants of surface complexes formed by adsorption reaction with SiO₂ and cations at pH 4 were calculated from the values of the MCSAs.     

Growth inhibition and induction of apoptosis in NB4 promyelocytic leukemia cells by trypsin inhibitor from sweet potato storage roots

The objective of this study was to investigate the antiproliferative effect and the mechanism of trypsin inhibitor (TI) from sweet potato [Ipomoea batatas (L.) Lam. 'Tainong 57'] storage roots on NB4 promyelocytic leukemia cells. The results showed that TI inhibited cellular growth of NB4 promyelocytic leukemia cells in a time-dependent and dose-dependent manner, and treatment for 72 h induced a marked inhibition of cellular growth, showing an IC50 of 57.1 +/- 8.26 microg/mL. TI caused cell cycle arrest at the G1 phase as determined by flow cytometric analysis and apoptosis as shown by DNA laddering. TI-induced cell apoptosis involved p53, Bcl-2, Bax, and cytochrome c protein in NB4 cells. P53 and Bax proteins were accumulated, and antiapoptotic molecule Bcl-2 was decreased in the tested cells in a time-dependent manner during TI treatment. TI also induced a substantial release of cytochrome c from the mitochondria into the cytosol. Hence, TI induced apoptosis in NB4 cells through a mitochondria-dependent pathway, which was associated with the activation of caspase-3 and -8. These results demonstrated that TI induces NB4 cell apoptosis through the inhibition of cell growth and the activation of the pathway of caspase-3 and -8 cascades.     

Antioxidant activity and inhibition of alpha-glucosidase by trans-resveratrol, piceid, and a novel trans-stilbene from the roots of Israeli Rumex bucephalophorus L

The roots of Rumex bucephalophorus, collected in Israel, were analyzed for trans-stilbenes. Two stilbene-O-glycosyl derivatives were identified, in addition to 3,5,4'-trihydroxystilbene (1) (resveratrol). The stilbene-O-glycosyl derivatives were 5,4'-dihydroxystilbene-3-O-beta-d-glucopyranoside (2) (piceid) and the new 5,4'-dihydroxystilbene-3-O-alpha-arabinopyranoside (3), which is being named rumexoid. The structure of rumexoid was elucidated by using spectroscopic data. The antioxidant capacities of stilbenoids 1-3 were determined and expressed as trolox equivalent antioxidant capacity (TEAC). TEAC value for trans-resveratrol was highest (2.7) and for rumexoid lowest (1.5). In vitro, trans-resveratrol and rumexoid demonstrated a potent inhibitory effect on alpha-glucosidase activity (IC50 < 0.1 and < 0.5 mM, respectively). The commercial antidiabetic agent acarbose was shown to inhibit only 35% of the enzyme activity at 0.5 mM. The addition of piceid to the reaction mixture did not inhibit alpha-glucosidase in vitro in the range of concentrations used. These findings extend the range of reported beneficial effects of stilbene derivatives, and demonstrate the multifaceted activities that dietary polyphenols may exert in the intestine, where their concentrations are highest in the body.     

Hydrolysis of naptalam and structurally related amides: inhibition by dissolved metal ions and metal (hydr)oxide surfaces.

In metal ion-free solutions, the secondary amide naptalam hydrolyzes more rapidly as the pH is decreased; intramolecular nucleophilic attack by a carboxylate side group is very likely involved. Millimolar levels of dissolved Cu(II) and Zn(II) inhibit hydrolysis between pH 3.6 and pH 6.5. Metal ion-naptalam complex formation is important since addition of the competitive ligand citrate lessens the inhibitory effect. The metal (hydr)oxide surfaces Al(2)O(3) and FeOOH inhibit naptalam hydrolysis to a lesser degree; inhibition is proportional to the extent of naptalam adsorbed. Secondary amides (propanil, salicylanilide, and N-1-naphthylacetamide) and tertiary amides (N-methyl-N-1-naphthylacetamide, furalaxyl, and N, N-diethylsalicylamide) that lack carboxylate side groups do not hydrolyze within 45 days of reaction, even when millimolar Cu(II) concentrations are present. Tertiary amides possessing carboxylate side groups (N,N-diethyl-3,6-difluorophthalamic acid and N, N-dimethylsuccinamic acid) do hydrolyze but are insensitive to the presence or absence of Cu(II). The inhibitory effect is believed to occur via metal coordination of (1) the carbonyl group of naptalam, which induces deprotonation of the amide group and makes the substrate less reactive toward nucleophilic attack; (2) the free carboxylate group of naptalam, which blocks intramolecular nucleophilic attack; or (3) a combination of the two.