Comparison of the metabolism and elimination of pyrilamine maleate in the rat,mouse and female rhesus monkey

Elimination and metabolic profiles of the O-glucuronide conjugated products of pyrilamine and their nonconjugated O-dealkylated and N-desmethyl pyrilamine products were determined after the oral administration of (¹⁴C)-pyrilamine maleate to Fischer 344 rats, B₆C₃F1 mice and female rhesus monkeys by stomach tube or i.v. The total cumulative urinary and fecal pyrilamine products were determined. The conjugated pyrilamine metabolites, isolated and identified were the glucuronide products of O-dealkylated pyrilamine and ring-hydroxylated pyrilamine, and the nonconjugated metabolites were predominately the N-desmethylpyrilamine and O-dealkylated pyrilamine and their ring-hydroxylated products. Statistically significant differences were observed in the percentages of the conjugated and nonconjugated metabolites of pyrilamine excreted by the three species studied.     

The influence of the rearing host on the response of the parasitoid Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) to odours from Ephestia kuehniella and Plodia interpunctella in a Y-tube olfactometer

Venturia canescens (Gravenhorst) is an ichneumonid generalist parasitoid that successfully attacks the larvae of different lepidopteran pests that infest stored products. These pest species include Plodia interpunctella and Ephestia kuehniella. In this study, we aimed to evaluate the influence of the rearing host on the parasitoid’s ability to detect and respond to a new host different from the rearing species. For this reason, the trials tested the preference of parasitoids reared on P. interpunctella or E. kuehniella for products that were or were not infested with larvae of these hosts. The trials were conducted in a Y-tube olfactometer. Regardless of the rearing host species, the parasitoids showed no preference for uninfested products. The parasitoids were attracted to products infested with larvae of their rearing host in preference to uninfested products. They also showed preferential attraction to products infested with the new host over uninfested products. E. kuehniella was the preferred host, irrespectively of the parasitoid host rearing species. The results are discussed to develop a better understanding of the ecology of V. canescens for its application in biological control.     

Auxin-induced changes in the population of translatable messenger RNA in elongating sections of soybean hypocotyl

In vitro translation products of polyadenylated RNA from untreated and auxin-treated elongating sections of soybean (Glycine max var. Wayne) hypocotyl were analyzed by two-dimensional polyacrylamide gel electrophoresis. The levels of translatable messenger RNA for at least ten in vitro translation products are increased by auxin treatment. The induction by auxin occurs rapidly (within 15 minutes), and the amounts of the induced in vitro translation products increase with time of auxin treatment. Indoleacetic acid has the same effect on the population of translatable messenger RNA as 2,4-dichlorophenoxyacetic acid. The auxin-induced in vitro translation products disappear rapidly when Actinomycin D is present during the last two hours of a three-hour auxin treatment.     

Alicyclobacillus Contamination in the Production Line of Kiwi Products in China

Alicyclobacillus are spoilage microbes of many juice products, but contamination of kiwi products by Alicyclobacillus is seldom reported. This study aims to investigate the whole production line of kiwi products in China to assess the potential risk of their contamination. A total of 401 samples from 18 commercial products, 1 processing plant and 16 raw material orchards were tested, and 76 samples were positive, from which 76 strains of microbes were isolated and identified as 4 species of Alicyclobacillus, including Alicyclobacillus acidoterrestris, Alicyclobacillus contaminans, Alicyclobacillus herbarius and Alicyclobacillus cycloheptanicus, and another 9 strains as 3 species of Bacillus by sequencing of their 16S rDNA. Through phylogenetic tree construction and RAPD-PCR amplification, it was found that there exist genotypic diversities to some extent among these isolates. Four test strains (each from one species of the 4 Alicyclobacillus species isolated in this study) could spoil pH adjusted kiwi fruit juice and some commercial kiwi fruit products with producing guaiacol (11–34 ppb).     

The simple and rapid detection of specific PCR products from bacterial genomes using Zn finger proteins

A novel method of rapid and specific detection of polymerase chain reaction (PCR) products from bacterial genomes using Zn finger proteins was developed. Zn finger proteins are DNA-binding proteins that can sequence specifically recognize PCR products. Since Zn finger proteins can directly detect PCR products without undergoing dehybridization, unlike probe DNA, and can double check the specific PCR amplification and sequence specificity of the PCR products, this novel method would be quick and highly accurate. In this study, we tried to detect Legionella pneumophila using Sp1. It was found that a 49 bp L. pneumophila-specific region containing the Sp1 recognition site is located on the flhA gene of the L. pneumophila genome. We succeeded in specifically detecting PCR products amplified from L. pneumophila in the presence of other bacterial genomes by ELISA, and demonstrated that Sp1 enables the discrimination of L. pneumophila-specific PCR products from others. By fluorescence depolarization measurement, these specific PCR products could be detected within 1 min. These results indicate that the rapid and simple detection of PCR products specific to L. pneumophila using a Zn finger protein was achieved. This methodology can be applied to the detection of other bacteria using various Zn finger proteins that have already been reported.     

Auxin- and ethylene-induced changes in the population of translatable messenger RNA in Basal sections and intact soybean hypocotyl

In vitro translation products of polyadenylated RNA from untreated and auxin-treated basal sections of soybean (Glycine max var. Wayne) hypocotyl were analyzed by two-dimensional polyacrylamide gel electrophoresis. Within one hour of 2,4-dichlorophenoxyacetic acid treatment, the translatable messenger RNAs for at least twelve in vitro translation products are modulated upward. In vitro translation products of polyadenylated RNA from untreated, auxin-treated and Ethephon-treated intact soybean hypocotyl were also analyzed. Within two hours of treatment with either 2,4-dichlorophenoxyacetic acid or Ethephon, the translatable messenger RNAs for a group of high molecular weight in vitro translation products are modulated upward. There is a particular set of translatable messenger RNA, encoding in vitro translation products in the 24,000 to 32,000 molecular weight range, that is specifically modulated upward by auxin treatment in intact soybean hypocotyl and in hypocotyl sections.     

Patterns of oligosaccharide production by polygalacturonases

The products of hydrolytic action of 18 enzyme preparations at pH 3·5 and 5·5 on pectate were analyzed by gel-filtration chromatography early in the course of reaction (8–15% hydrolysis), and at a time 10 times that required for 10% hydrolysis. The degree of hydrolysis at the latter time varied from 25 to 74%. Three patterns of oligosaccharide production could be distinguished: endo-hydrolysis, exo-hydrolysis, and that due to S-polygalacturonase. The initial products of endo-hydrolysis were mixed oligosaccharides 5–30 units long; monomer and dimer appeared early but represented less than 2% of the products until late in the reaction. exo-Polygalacturonase (not entirely free of endo-) showed predominant production of the monomer and was clearly evident when mixed with four parts of endo-polygalacturonase. The time course of reducing group production by highly purified S-polygalacturonase could be reproduced by the above mixture of exo- and endo-polygalacturonases, but the pattern of products and the pH relations could not. The initial products of S-polygalacturonase were monomer, dimer and pentamer with lesser amounts of trimer and tetramer. After the hydolysis of the polymer and large oligomers, the pentamer was attacked by S-polygalacturonase, in the same way that the accumulated hexamer, etc. were finally hydrolysed by the endo-polygalacturonase.     

Flavour biogenesis. Partial purification and properties of a fatty acid hydroperoxide cleaving enzyme from fruits of cucumber

A membrane-bound enzyme, which catalyses the cleavage of fatty acid hydroperoxides to carbonyl fragments, has been partially purified from cucumber fruit. The isomeric 9- and 13-hydroperoxydienes (but not the hydroxydienes) derived from both linoleic and linolenic acids are cleaved by the enzyme but a mixture of 9- and 10-hydroperoxymonoenoic derivatives of oleic acid was not attacked. No evidence was obtained for free intermediates between fatty acid hydroperoxides and the cleavage products. Major volatile products were: cis-3-nonenal and hexanal (from 9- and 13-hydroperoxides of linoleic acid respectively) or cis-3,cis-6-nonadienal and cis-3-hexenal (from 9- and 13-hydroperoxides of linolenic acid). The increase in the ratio of cis-3- to trans-2-enal products with enzyme purification indicated that cis-3-enals are the immediate cleavage products and that the trans-2- forms are produced by subsequent isomerization.     

Evaluation of persistence and relative toxicity of some pest control products to adults of two native trichogrammatid species in Kenya

The utilization of native trichogrammatids for biocontrol of Helicoverpa armigera (Hbn.) and their potential integration with pesticide use are currently receiving attention. In this study the interaction of adults of Trichogramma sp. nr. mwanzai and Trichogrammatoidea sp. nr. lutea with commonly used pesticide products was investigated. The toxicity of eight pest control products commonly used in vegetable crops in Kenya, were evaluated by exposing the adults of the two species to detached potted tomato leaves at different intervals after spraying. Two biologically derived products, Achook^? (neem-based) and Dipel^? (Bt ssp. kurstaki)—were found to be harmless and had no persistent toxicity on both the trichogrammatid species. Two organophosphate products tested, dimethoate (Rogor^?) and malathion (Malathion^?) were found to be ‘slightly harmful’ and ‘moderately persistent’ respectively. Three other synthetic insecticides, lambdacyhalothrin (Karate^?), bifenthrin (Brigade^?) and alpha-cypermentrin (Fastac^?) were found to be ‘moderately harmful’ and ‘persistent’ respectively. Polytrin^? (mixture of cypermethrin and profenofos) was also found to be persistent but only ‘slightly harmful’. On the basis of five concentrations tested (0.032, 0.016, 0.008, 0.004 and 0.002 of the recommended field rates) the LC₅₀ values for adult T. sp. nr. mwanzai were estimated as 285, 411, 435 and 1,916 (μg active ingredient (a.i) ml^?1) for dimethoate; malathion; lambdacyhalothrin as well as cypermethrin?+?profenofos respectively. The corresponding values for T. sp. nr. lutea were 247, 251, 278 and 697 respectively. Further, Karate^? appeared to be the least toxic among the four products, across all the respective concentrations. The study was an attempt to adopt the methodologies of the IOBC (International Organization for Biological Control) on non-target risk assessment for pest control products to cater for the local needs of integrating the use of the trichogrammatids along with other pest control products.     

Methylation of DNA by 3H-14C-methyl-labelled N-methyl-N-nitrosourea--evidence for transfer of the intact methyl group

The following products have been isolated from methyl-labelled N-methyl-N-nitrosourea (MNUA) and DNA after reaction at pH 8: 1-, 3-, 7-methyladenines, 3-methyldeoxycytidine, 3-, 7- and O⁶-methylguanines, 3- and O⁴-methylthymidines. Comparison of C³H₃ and ¹⁴CH₃ labelling showed that the ratio ${}^3\text{H}{}^{14}\text{C}$ in the products was the same and equal to that in the original reagents, in accord with the concept that the methyl group is transferred intact, and not via diazomethane. In some cases, most notably with O⁶-methyldeoxyguanosine, the ³H-labelled products were found to elute from Dowex 50 (NH₄⁺ form) slightly ahead of ¹⁴C-labelled or unlabelled products.     

An unexpected response of Torulopsis glabrata fusion products to x-irradiation

Intra-species fusion products of Saccharomyces cerevisiae, Saccharomyces unisporus and Torulopsis glabrata have been isolated following polyethylene glycol-induced fusion of protoplasts and selection for prototrophic colonies. Staining with lomofungin showed that all fusion products were uninucleate. Measurement of DNA content mostly gave values between haploid and diploid levels indicating that the majority of fusion products were aneuploid. Nevertheless fusion products of S. cerevisiae and S. unisporus were, as expected, more resistant to X-irradiation than their haploid parents. By contrast, the X-ray doze—response curve of all T. glabrata fusion products was indistinguishable from their progenitors despite the fact that mitotic segregants could be recovered amongst the survivors to X-rays. A possible explanation for the behaviour towards X-rays of T. glabrata fusion products is that this species lacks a DNA repair pathway involving recombination between homologous chromosomes. We conclude from this study that the shape of the X-ray dose—response curve should not be taken to indicate the ploidy of new yeast isolates without supporting data.     

Comparison of Fc N-Glycosylation of Pharmaceutical Products of Intravenous Immunoglobulin G

Intravenous immunoglobulin (IVIg) products from different pharmaceutical companies vary in composition, in part because of the selected blood donors and production process. N-glycosylation of the Fc-portion of IgG varies between blood donors and may influence both the side-effects and therapeutic effectiveness of IVIg. At present, the variation in Fc N-glycosylation between IVIg products has not been defined. Utilizing mass spectrometry, we performed relative quantitation of the Fc N-glycosylation of IgG, assessing a total of 154 unique lot numbers of IVIg. Seven products showed comparable Fc N-glycosylation, with only one product differing from the others in all glycosylation features (galactosylation, sialylation, fucosylation and bisecting N-acetylglucosamine). However, the mean difference did not exceed 3%. Within product variation was present to a minor degree, but largely indistinguishable from analytical variation. In conclusion, we expect that the minor variation in Fc N-glycosylation between IVIg products has a small effect, if any, on the biological activity.     

The Effects of Glucosinolates and Their Breakdown Products on Necrotrophic Fungi

Glucosinolates are a diverse class of S- and N-containing secondary metabolites that play a variety of roles in plant defense. In this study, we used Arabidopsis thaliana mutants that contain different amounts of glucosinolates and glucosinolate-breakdown products to study the effects of these phytochemicals on phytopathogenic fungi. We compared the fungus Botrytis cinerea, which infects a variety of hosts, with the Brassicaceae-specific fungus Alternaria brassicicola. B. cinerea isolates showed variable composition-dependent sensitivity to glucosinolates and their hydrolysis products, while A. brassicicola was more strongly affected by aliphatic glucosinolates and isothiocyanates as decomposition products. We also found that B. cinerea stimulates the accumulation of glucosinolates to a greater extent than A. brassicicola. In our work with A. brassicicola, we found that the type of glucosinolate-breakdown product is more important than the type of glucosinolate from which that product was derived, as demonstrated by the sensitivity of the Ler background and the sensitivity gained in Col-0 plants expressing epithiospecifier protein both of which accumulate simple nitrile and epithionitriles, but not isothiocyanates. Furthermore, in vivo, hydrolysis products of indole glucosinolates were found to be involved in defense against B. cinerea, but not in the host response to A. brassicicola. We suggest that the Brassicaceae-specialist A. brassicicola has adapted to the presence of indolic glucosinolates and can cope with their hydrolysis products. In contrast, some isolates of the generalist B. cinerea are more sensitive to these phytochemicals.     

Oxidative breakdown and conversion of urocanic acid isomers by hydroxyl radical generating systems

cis-Urocanic acid (cis-UCA), formed from trans-urocanic acid (trans-UCA) by photoisomerization, has been shown to mimic suppressive effects of UV on the immune system. It is our hypothesis that UCA oxidation products in the skin play a role in the process of immunosuppression. Recently, both UCA isomers were found to be good hydroxyl radical scavengers and in this context we investigated the formation of products resulting from the interaction of hydroxyl radicals with UCA. Hydroxyl radicals were generated by (1) UV/H₂O₂ (photooxidation), (2) ferrous ions/H₂O₂ (Fenton oxidation) and (3) cupric ions/ascorbic acid. Oxidation products were identified by spectrometric methods and assessed by reversed-phase HPLC analysis. The photooxidation of UCA was induced by UV-B and UV-C, but not by UV-A radiation. Photooxidation and Fenton oxidation of trans-UCA, as well as of cis-UCA yielded comparable chromatographic patterns of UCA oxidation products. Several of the formed products were identified. The formation of three identified imidazoles was shown in UV-B exposed corneal layer samples, derived from human skin.     

Proteomic Analysis of Excretory-Secretory Products of Mesocestoides corti Metacestodes Reveals Potential Suppressors of Dendritic Cell Functions

Accumulating evidences have assigned a central role to parasite-derived proteins in immunomodulation. Here, we report on the proteomic identification and characterization of immunomodulatory excretory-secretory (ES) products from the metacestode larva (tetrathyridium) of the tapeworm Mesocestoides corti (syn. M. vogae). We demonstrate that ES products but not larval homogenates inhibit the stimuli-driven release of the pro-inflammatory, Th1-inducing cytokine IL-12p70 by murine bone marrow-derived dendritic cells (BMDCs). Within the ES fraction, we biochemically narrowed down the immunosuppressive activity to glycoproteins since active components were lipid-free, but sensitive to heat- and carbohydrate-treatment. Finally, using bioassay-guided chromatographic analyses assisted by comparative proteomics of active and inactive fractions of the ES products, we defined a comprehensive list of candidate proteins released by M. corti tetrathyridia as potential suppressors of DC functions. Our study provides a comprehensive library of somatic and ES products and highlight some candidate parasite factors that might drive the subversion of DC functions to facilitate the persistence of M. corti tetrathyridia in their hosts.     

Assembly of bacteriophage T4 tail fibers. IV. Subunit composition of tail fibers and fiber precursors

Using a novel purification procedure, the protein composition of the tail fibers of bacteriophage T4 has been determined. Fibers contain four proteins whose molecular weights, as estimated by sodium dodecyl sulfate/acrylamide gel electrophoresis, are 150,000, 125,000, 40,000 and 24,000. The two largest proteins have been previously identified as the products of genes 34 (P34) and 37 (P37), respectively (King and Laemmli, 1971; Ward and Dickson, 1971). The two smaller proteins have now been identified as the products of genes 35 (P35) and 36 (P36), respectively. The products of the two other known phage genes required for fiber assembly, 38 and 57, have been identified as non-structural phage proteins with molecular weights of 26,000 and 10,000, respectively.     

Presence of pathogenic Vibrio Parahaemolyticus in waters and seafood from the Tunisian Sea

The occurrence of the hemolysin genes, tdh and trh, in Vibrio parahaemolyticus strains isolated from environmental samples collected from various exported seafood products comprising of fishes and shellfish (Mytilus edulis and Crassostrea gigas) or seawater, was studied. Eight strains were confirmed as V. parahaemolyticus by toxR -based polymerase chain reaction and only one strain out of these 8 strains was positive for tdh and trh genes. Toxigenic V. parahaemolyticus isolates are present in Tunisian coastal areas and they may also be present in Tunisian exported seafood products.     

3-Hydroxypropylglucosinolate,a new glucosinolate in seeds of Erysimum hieracifolium and Malcolmia maritima

Glucosinolates from seed meals of Erysimum hieracifolium and Malcolmia maritima were treated with thioglucosidase (EC 3.2.3.1), and the resultant aglucon products investigated. A major product from E. hieracifolium was 3-hydroxypropyl isothiocyanate from the novel precursor 3-hydroxypropylglucosinolate. Other aglucon products were 3-methylsulfonylpropyl, 3-methylsulfinylpropyl, 3-methylthiopropyl, and allyl isothiocyanates. The aglucon products from M. maritima seed meal included 3-hydroxypropyl isothiocyanate, in addition to the previously known 3-methylsulfonylpropyl and 3-benzoyloxypropyl isothiocyanantes.     

Mechanism of Reactive Orange 16 degradation with the white rot fungus Irpex lacteus

The study focuses on the Reactive Orange 16 (RO16) degradation products obtained by fungal treatment of the dye. Eighty percent of dye decolorization was achieved within 24 h by Irpex lacteus cultures immobilized on polyurethane foam (PUF). Dye degradation products were investigated using LC–MS analysis. Three compounds were identified as the dye intermediates: 6-acetamido-3,4-dioxo-3,4-dihydronaphthalene-2-sulfonate (m/z 294), (E)-2-(4-acetamidophenyl)-1-carboxyethenesulfonate (m/z 284), and 4-(2-hydroxyethylsulfonyl)phenolate (m/z 201). Despite significant laccase activities detected in the fungal cultures, no backward polymerization of the reaction products resulting in recurrent colorization was observed after fungal treatment of the dye solution.