Decay study of pesticide residues in apple samples

A method for monitoring pesticides in apple samples, including Soxhlet extraction, an evaporation step and capillary gas chromatography with nitrogen-phosphorus detection, is applied to a decay study of the carbamate pesticide ethiofencarb. The evaporation step is carried out by a surface nitrogen flow and the recoveries of ethiofencarb measured in standard solutions of ethyl acetate and apple extracts. The results of the decay studies show that ethiofencarb is degraded in the apple peel faster than in the interior of the fruit. The methodology is then applied to decay studies in the laboratory of the organophosphorus pesticides, diclofluanid, fenitrothion and malathion.     

A carbamate analogue of amsacrine with activity against non-cycling cells stimulates topoisomerase II cleavage at DNA sites distinct from those of amsacrine

AMCA (methyl N-[4-(9-acridinylamino)-2-methoxyphenyl]carbamate hydrochloride), an amsacrine analogue containing a methylcarbamate rather than a methylsulphonamide side chain, contrasts with amsacrine, doxorubicin and etoposide in its relatively high cytotoxicity against non-cycling tumour cells. AMCA bound DNA more tightly than amsacrine, but the DNA base selectivity of binding, as measured by ethidium displacement from poly[dA-dT].[dA-dT] and poly[dG-dC].[dG-dC], was unchanged. AMCA-induced topoisomerase cleavage sites on pBR322, C-MYC and SV40 DNA were investigated using agarose or sequencing gels. DNA fragments were end-labelled, incubated with purified topoisomerase II from different mammalian sources and analysed after treatment with sodium dodecylsulphate/proteinase K. AMCA stimulated the cleavage activity of topoisomerase II, but the DNA sequence selectivity of cleavage was different from that of amsacrine and other topoisomerase inhibitors. It was similar to that of the methoxy derivative of AMCA, indicating that the changed specificity resulted from the carbamate group rather than from the methoxy group. The pattern of DNA cleavage induced by AMCA was similar for topoisomerase II alpha and II beta.     

Bioavailability and effect of pirimicarb on Daphnia magna in a laboratory freshwater/sediment system

In tests with newborn and one-week-old Daphnia magna, 48-h EC50 values of 21-24 microg/L and 16 microg/L pirimicarb, respectively, were found. Older animals thus were as sensitive to pirimicarb as newborn animals. In an experiment with sediment included in the test system, all mother animals survived for 72 h at 20 microg/L, and the number of offspring was not reduced relatively to the control. Addition of sediment thus reduced the toxicity of pirimicarb toward Daphnia magna. Pirimicarb was accumulated 12-16 times (5-7% of total) in the sediment, but the water concentrations of pirimicarb were not reduced significantly during the experiment, due to the small amount of sediment used. Accumulation in the sediment was found independent of the water concentration used. This was also the case with bioaccumulation in Daphnia magna, where a bioaccumulation factor of 31-37 was found on a dry weight basis. In water without sediment a BCF of 50 was found. Addition of sediment also reduced the accumulation of pirimicarb in the daphnids. The reduced bioavailability of pirimicarb may derive from humic acid and related compounds released from the sediment.     

Dynamic kinetic asymmetric cycloadditions of isocyanates to vinylaziridines

The first examples of the use of racemic vinylaziridines in a Pd-catalyzed dynamic kinetic asymmetric transformation have been examined. Optimization studies of the Pd-catalyzed addition of vinylaziridines to isocyanates revealed that the chiral ligand between trans-1,2-diaminocyclohexane and 2-diphenylphosphino-1-naphthoic acid is superior to that involving 2-diphenylphosphino benzoic acid. Surprisingly, high ee's required the use of an acid whose pKa was about 4.7 +/- 0.1 as a cocatalyst. Both acetic acid and hydroxybenzotriazole meet this requirement. Less electrophilic isocyanates (e.g., benzyl, p-methoxyphenyl) gave higher ee's than more electrophilic ones (phenyl or benzoyl). Both N-benzyl and N-arylaziridines react well to give good yields and ee's, whereas N-tosylaziridines gave lower ee's. A 1,1-disubstituted aziridine led to the formation of a tertiary C-N bond with ee's comparable to the formation of the secondary C-N bond. The products were easily reduced almost quantitatively to the sensitive imidazolidines which can be readily hydrolyzed to the vicinal diamines. The reactivity pattern is consistent with a Curtin-Hammett situation wherein the enantiodiscriminating event is the cyclization of a rapidly equilibrating dynamic pi-allyl palladium intermediate.     

Lack of relay toxicity in ferret hybrids fed carbaryl-treated prairie dogs

Carbaryl (1-napthol methylcarbamate) is being considered for control of fleas on prairie dogs (Cynomys spp.) used in black-footed ferret (Mustela nigripes) recovery in the western United States. The potential for relay toxicity in ferrets was determined by feeding carbaryl treated prairie dogs to black-footed ferret x Siberian polecat (M. eversmanni) hybrids. Adult prairie dogs were treated topically with 2.5 g of commercial 5% carbaryl dust sold as flea powder. After 14 days prairie dogs were killed and fed to ferrets. Potential for relay toxicity was evaluated by analyzing ferret blood cholinesterase (CHe), prairie dog brain Che, and hepatic carbamate concentration. There was no difference between pre- and post-exposure blood CHe activity, nor did treated prairie dog brain CHe differ significantly from controls. Post-exposure blood CHe did not exhibit reactivation after dilution in aqueous buffer. Hepatic carbaryl concentrations were less than detection limits (50 ppb). Based on these results, we conclude that short-term use of carbaryl for flea control on prairie dogs does not pose a hazard of relay toxicity in black-footed ferrets.     

Synthetic macromolecular inhibitors of human leukocyte elastase. 2. Effect of loading of a peptidyl carbamate inhibitor and molecular size of polymer backbone on its inhibitory capacity

Several macromolecular inhibitors of human leukocyte elastase (HLE) were prepared by covalently bonding a low molecular weight HLE inhibitor peptidyl carbamate, p-nitrophenyl-N-[succinyl-L-alanyl-L-ananyl-L-prolylmethyl]- N-isopropyl carbamate 1, with the neutral hydrophilic polymer, poly-alpha,beta-[N-(2-hydroxyethyl)-D,L-aspartamide], PHEA 2. These novel polymeric compounds differed in the molecular size of their PHEA polymer backbone and the extent of loading of the peptidyl carbamate, (PC). They were shown to efficiently inhibit HLE (Ki = 97 to 12.8 nM) as intact macromolecular entities and were found to be more stable to hydrolysis than the non-polymer bound low molecular weight inhibitor 1. The inhibition of HLE by the novel macromolecular inhibitors was found to be noncompetitive and reversible, proceeding via slow formation of inhibitor-enzyme complex. The effect of loading of 1 and molecular size of the PHEA 2 polymer on enzymatic parameters Ki, kon and koff is discussed and a possible mechanism of inhibition is presented.     

A tale of novel intoxication: seven cases of gamma-hydroxybutyric acid overdose

The chemical behaviours of 4-methyl-2-oxo-2H-benzopyran-7-yl oxoacetyl hydrazine (2) towards some different reagents such as anhydride compounds, aromatic aldehydes, carbon disulphide, and nitrous acid yielded the corresponding pathalazine derivatives (3, 4, 5), hydrazone derivative (6), 1,3,4-oxadiazole derivative (7, 8, 9) and acid azide (10) respectively. Treatment of 10 with absolute alcohols, amines and ethyl amino acid ester gave the corresponding carbamate derivative (11), substituted urea derivative (12) and ethyl substituted alkyl acetate (13) respectively. The biological activity of some synthesized compounds was evaluated.     

Snapshot of a phosphorylated substrate intermediate by kinetic crystallography

The ATP-dependent enzyme dethiobiotin synthetase from Escherichia coli catalyses the formation of dethiobiotin from CO2 and 7, 8-diaminopelargonic acid. The reaction is initiated by the formation of a carbamate and proceeds through a phosphorylated intermediate, a mixed carbamic phosphoric anhydride. Here, we report the crystal structures at 1.9- and 1.6-A resolution, respectively, of the enzyme-MgATP-diaminopelargonic acid and enzyme-MgADP-carbamic-phosphoric acid anhydride complexes, observed by using kinetic crystallography. Reaction initiation by addition of either NaHCO3 or diaminopelargonic acid to crystals already containing cosubstrates resulted in the accumulation of the phosphorylated intermediate at the active site. The phosphoryl transfer step shows inversion of the configuration at the phosphorus atom, consistent with an in-line attack by the carbamate oxygen onto the phosphorus atom of ATP. A key feature in the structure of the complex of the enzyme with the reaction intermediate is two magnesium ions, bridging the phosphates at the cleavage site. These magnesium ions compensate the negative charges at both phosphate groups after phosphoryl transfer and contribute to the stabilization of the reaction intermediate.     

Skin sensitization to linalyl hydroperoxide: support for radical intermediates

In order to better understand the skin sensitization mechanism of allylic hydroperoxides, linalyl hydroperoxide (1) and several of its potential rearrangement products-epoxylinalool (2), epoxynerol (3), epoxygeraniol (4), and furan (5) and pyran (6) derivatives-were synthesized. The sensitizing properties of these molecules have been screened on mice using the local lymph node assay (LLNA) and further evaluated on guinea pigs using the Freund's complete adjuvant test (FCAT). Linalyl hydroperoxide (1) and linalyl epoxide (2) were found to be sensitizers, while the other compounds were classified as mild sensitizers or nonsensitizers. In the guinea pigs, no cross-reactions were observed between skin sensitizers 1 and 2. Radical-trapping experiments were carried out on linalyl hydroperoxide (1) using TTBP as trapping agent and Fe(3+)-TPP as radical inducer. The major reaction taking place is the formation of a furan ring by intramolecular reaction of the oxygen-centered radical with the isoprenyl double bond with the formation of a tertiary radical. Reaction of this intermediate with radicals derived from TTBP gave compounds 10a,b in 25% yield. The second important reaction, accounting for 14%, is taking place on the allylic double bond with the formation of a less stable primary radical which is not trapped by a TTBP-derived radical but by a hydroxy radical to give a mixture of epoxides 3 and 4. These results are in favor of the formation of a carbon-centered reactive radical as intermediate in the skin sensitization to linalyl hydroperoxide.     

1997 Sir William Refshauge Lecture. Skeletal muscle glucose metabolism during exercise: implications for health and performance

The shear bond strengths of an amalgam (Permite C) and a gallium alloy (Galloy) to dentin, mediated by four dentin adhesives (Super-Bond D-Liner, Super-Bond D-Liner II, Paama 2, and Panavia 21), were investigated. Flat labial dentin surfaces were prepared from bovine lower incisor teeth. A 3 mm-in-diameter area of dentin was bonded according to each manufacturer's directions before placement of Permite C or Galloy. The bonds were stressed in shear at a crosshead speed of 1 mm/min. The mean shear bond strengths were analyzed using one-way ANOVA and Student's t-test, and fracture modes were assessed under X20 magnification and analyzed using Kruskal-Wallis and Mann-Whitney U tests. Scanning electron micrographs were taken of the bond interface of separate samples. The results showed no significant difference among the bond strengths of Super-Bond D-Liner (2.79 MPa, 2.69 MPa), Super-Bond D-Liner II (3.41 MPa, 2.65 MPa), and Paama 2 (0.70 MPa, 0.50 MPa) bonded to Permite C and Galloy (respective values in parentheses); however, Panavia gave a significantly better bond with Permite C (0.42 MPa) than with Galloy (0 MPa). Super-Bond D-Liner and Super-Bond D-Liner II gave stronger bonds than Paama 2 and Panavia with both Permite C and Galloy. For each dentin adhesive, there was no difference in fracture mode between Permite C and Galloy. It was concluded that, since all bond strengths were very low, none of the dentin adhesives tested would enhance the clinical retention of Permite C or Galloy. However, although the use of Paama 2 with Galloy was originally recommended by the manufacturer for dentin sealing purposes, no adhesion was claimed.     

Effects of N-methacryloyl amino acid applications on hybrid layer formation at the interface of intertubular dentin

To understand the role of NMAA in the bonding of composite resin to a dentin surface, we investigated the effects of N-methacryloyl amino acid (NMAA) application on the expansion of aggregated collagen fibers, formation of a hybrid layer, and the tensile bond strength between composite resin and dentin. Four NMAA derivatives--N-methacryloyl-alpha-glycine (NMGly), N-methacryloyl-gamma-amino n-butyric acid (NMBu), N-methacryloyl-alpha-hydroxyproline (NMHPro), and N-methacryloyl-alpha-glutamic acid (NMGlu)--were prepared and applied to dentin surfaces which had been etched with 40% by mass H3PO4 and air-blown. The shrunken collagenous layer expanded by approximately 50% to 70% by volume of the original collagenous layer thickness after application of the NMAA primers. Application of the bonding agent and composite resin after NMAA treatment resulted in the formation of a hybrid layer. The thickness of the hybrid layer was somewhat smaller than the collagenous layer formed by the NMAA treatment only, regardless of the type of NMAA used. The thickness of the hybrid layer was approximately ten times larger than that formed without NMAA treatment. Although all NMAA primers formed hybrid layers of similar thickness, higher tensile bond strengths, from 13 to 15 MPa, were obtained when etched and air-blown dentin was treated with NMBu, NMGly, or NMGlu. NMHPro gave only 6.6 MPa, a value similar to that obtained when no NMAA was used. We concluded, therefore, that formation of the hybrid layer is a necessary but insufficient condition for high bond strength.     

Dementia improvement with cytotoxic chemotherapy. A case of Alzheimer disease and multiple myeloma

Sperm transfer into the epididymis was completed without a blood meal, when newly emerged male cat fleas. Ctenocephalides felis (Bouché), were exposed to filter papers treated with juvenile hormone III or the juvenile hormone mimics fenoxycarb, methoprene, or pyriproxyfen. As the concentration of juvenile hormone or the time of flea exposure to juvenile hormone or the juvenile hormone mimics increased, the percentage of fleas that transferred sperm also increased. The percentage of pyriproxyfen-treated males that transferred sperm reached 100% after 3 d: whereas, 7 d exposure to juvenile hormone, fenoxycarb and methoprene was required for 100% of the males to transfer sperm. Although sperm were present in the epididymis of treated fleas, insemination of females did not take place off the host either on juvenile hormone-treated filter paper or on juvenile hormone-treated dog hair.     

Synthesis and biological activity of a 1 alpha,25-dihydroxyvitamin D2 analog bearing an amide group in the side-chain

Synthesis of a 1 alpha,25-dihydroxyvitamin D2 analog (3), in which the double bond in the side-chain is replaced by an amide group, is described. Condensation of a carboxylic acid (8) with an amine (6) gave an amide (9), which in turn led to 3 via several steps. The analog (3) could not bind to the chick cytosol vitamin D receptor, which indicated the importance of the hydrophobic interaction of the C(22)-C(23) double bond in 1 alpha,25-dihydroxyvitamin D2 (2) with the vitamin D receptor.     

Characteristic of Bond Zone between Powder Surfacing Layer and Base Metal

The characteristic of the bond zone between Ni-based alloy light beam surfacing layer(SL) and base metal (BM) was investigated by scanning electron microscope, energy dispersive spectrometer and X-ray diffraction. The results show that the bond zone, which consists of γ-Ni or γ- (Fe, Ni) planar crystal band close to SL and α-Fe bright band close to heat affected zone (HAZ), is actually the transition zone of composition and microstructure between SL and HAZ, and the metallurgical bond interface lies between the α-Fe bright band and HAZ. With the increase of light beam heat input from 2 kJ/mm to 4 kJ/mm, the width of the bond zone increases from 4μm to 15μm, and the morphology of bond interface changes from zigzag to straight. The formation of bond interface indicates the formation of reliable metallurgical bond between SL and BM.     

Jack bean urease (EC 3.5.1.5). V. On the mechanism of action of urease on urea, formamide, acetamide, N-methylurea, and related compounds

Acetamide and N-methylurea have been shown for the first time to be substrates for jack bean urease. In the enzymatic hydrolysis of urea, formamide, acetamide, and N-methylurea at pH 7.0 and 38 degrees C, kcat has the values 5870, 85, 0.55, and 0.075 s-1, respectively. The urease-catalyzed hydrolysis of all these substrates involves the active-site nickel ion(s). Enzymatic hydrolysis of the following compounds could not be detected: phenyl formate, p-nitroformanilide, trifluoroacetamide, p-nitrophenyl carbamate, thiourea, and O-methylisouronium ion. In the enzymatic hydrolysis of urea, the pH dependence of kcat between pH 3.4 and 7.8 indicates that at least two prototropic forms are active. Enzymatic hydrolysis of urea in the presence of methanol gave no detectable methyl carbamate. A mechanism of action for urease is proposed which involves initially an O-bonded complex between urea and an active-site Ni2+ ion and subsequently an O-bonded carbamato-enzyme intermediate.     

Mechanism of bond zone wave formation in explosion-clad metals

Results of experiments in which the collision variables were carefully controlled showed that bond zone wave formation during explosion cladding is analogous to the formation of vortex streets in fluid flow around an obstacle or in the collision of liquid streams. The fluid flow analogy explains the observed transition from a smooth metal-to-metal bond zone to a wavy bond zone above a critical collision velocity. This model is capable of predicting the minimum collision velocity necessary for bond zone wave formation in different metal systems and it also predicts correctly the strong dependence of wave size on collision angle. The magnitude of the wave size agrees with that predicted from fluid flow past a flat plate. Two other mechanisms of bond zone wave formation were explored experimentally and found to be invalid.     

Pre-steady-state kinetics of nucleotide insertion following 8-oxo-7,8-dihydroguanine base pair mismatches by bacteriophage T7 DNA polymerase exo-

8-Oxo-7,8-dihydroguanine (8-oxoGua) can base pair with either cytosine (C) or adenine (A) when replicated by DNA polymerases. The 8-oxoGua.A mismatch is extended in preference to the 8-oxoGua.C pair. Using a model 25-mer/36-mer DNA duplex containing either guanine (Gua).C, 8-oxoGua.C, or 8-oxoGua.A base pairs at the primer terminus and A at the standing start position, we found that the pre-steady-state addition of dTTP opposite A following all three base pairs by bacteriophage T7 DNA polymerase exo- showed burst kinetics, suggesting that extension of all three base pairs is controlled by the rate of a step at or before phosphodiester bond formation. Substitution of dTTP alpha S for dTTP yielded modest thio effects of 1-6, suggesting that extension of all three pairs is limited by the rate of the conformational change prior to phosphodiester bond formation. Pre-steady-state values for kpol (maximum polymerization rate) were 120, 12, and 28 s-1, and Kd values were 2, 75, and 22 microM for insertion of dTTP following Gua.C, 8-oxoGua.C, and 8-oxoGua.A base pairs, respectively. Additional analysis of extension was provided by substitution of A in the standing start position by 2-aminopurine (2-AP), a fluorescent base analogue. Comparison of rapid-quench gel-based assays with stopped-flow fluorescence quenching assays suggested that during addition of dTTP opposite 2-AP phosphodiester bond formation was rate-limiting when 8-oxoGua.C or 8-oxoGua.A were the preceding base pairs, while conformational change was rate-limiting when Gua.C was the preceding base pair. Furthermore, the difference in apparent conformational change rates for addition of dTTP opposite 2-AP following the 8-oxoGua base pairs was greater than the differences in their phosphodiester bond formation rates, suggesting that discrimination in extension may be influenced more by conformational change rates than the rates of phosphodiester bond formation in this mispaired system.     

Mass spectra of acetylenic fatty acid methyl esters and derivatives

A series of isomeric methyl octadecynoates was analyzed by mass spectrometry; each isomer gave a unique spectrum. The characteristic ions were those resulting from a McLafferty rearrangement of the allenic sites or of the already-rearranged allenic sites. The acetylenic esters were also subjected to oxymercuration whereupon a carbonyl group was formed at either of the original actylenic carbon atoms providing two oxostearates. Further reaction with NaBH4 formed hydroxy esters which, after silylation, gave diagnostic mass spectra indicative of the triple bond location. Applied to esters with both double and triple bonds, this procedure permitted differentiation between the two types of unsaturation. Methoxyl groups marked the original double bond locations and hydroxyls did so for triple bonds.     

An unusual case of diabetic cellulitis due to Pasturella multocida

Exposure of male Balb/C mice to mainstream cigarette smoke for 4 months, starting 10 or 30 days before the administration of ethyl carbamate (0.3% in drinking water for 3 weeks), resulted in an up to 57.6% (P < 0.05) decrease of lung adenoma multiplicity. However, the number of ethyl carbamate-induced lung tumors was not significantly affected by exposure to cigarette smoke when ethyl carbamate was injected i.p. in single doses of 0.5 or 1.0 g/kg, irrespective of the different treatment schedules used, i.e. (a) 10 days before and 4 days after the ethyl carbamate injection; (b) throughout the experiment starting 10 days before the ethyl carbamate injection, and (c) until the end of the experiment, starting 30 days after the ethyl carbamate injection. Disulfiram (500 mg/kg), given by gavage 24 h and 1 h before the ethyl carbamate injection, decreased by 88.5% (P < 0.001) the multiplicity of lung adenomas but had no effect on tumorigenesis when administered after the carcinogen injection. Proadifen (SKF-525 A, 50 mg/kg) injected i.p. 24 h and 1 h before and 24 h and 48 h after the injection with ethyl carbamate tended to decrease the multiplicity of lung adenomas, but not to a significant extent. Furthermore, disulfiram given 24 h and 1 h before the i.p. administration of ethyl carbamate completely prevented its clastogenicity in mouse bone marrow. On the other hand, cigarette smoke, which was per se a weak clastogen in bone marrow erythroblasts, synergistically potentiated the clastogenic response to ethyl carbamate in a more than additive fashion.