Tick salivary gland extract-activated transmission of Borrelia afzelii spirochaetes

Saliva-activated transmission of Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 was demonstrated using salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks and C3H mice. Injection of Borrelia spirochaetes together with SGE increased the level of bacteraemia and accelerated the appearance of bacteria in the urinary bladder, compared with the injection of spirochaetes alone. More I. ricinus nymphs became infected when feeding on mice inoculated with B. afzelii plus SGE. Analysis of cytokines produced by cells of draining lymph nodes from SGE-treated mice showed a suppression of proinflammatory cytokines IFN-gamma, IL-6 and GM-CSF following a transient upregulation in comparison with the control mice infected without SGE.     

Induction of glucokinase mRNA by dietary phenolic compounds in rat liver cells in vitro

Diabetes and its complications, including oxidative stress, are major reasons for medical intervention and one of the most frequent causes of death in developed countries. Several lines of data suggest that the use of certain dietary polyphenolic compounds may alter glucose metabolism, thus decreasing the risk for type 2 diabetes. In this paper, we present the effect of phenolic acids (caffeic, chlorogenic, rosmarinic, and ferulic) and extracts from Smallanthus sonchifolius and Prunella vulgaris on glucose production in rat hepatocytes and on glucokinase, glucose-6-phosphatase, and phosphoenol-pyruvate carboxykinase mRNA expression in rat hepatoma Fao cells. The phenolics at 500 microM and after 1 h incubation lowered glucose production via both gluconeogenesis (10 mM alanine or dihydroxyacetone as precursors) and glycogenolysis compared with metformin. Most of the phenolics increased the level of glucokinase mRNA after 24 h in the same way as insulin (10(-7) M).     

HPLC analysis of rosmarinic acid in feed enriched with aerial parts of Prunella vulgaris and its metabolites in pig plasma using dual-channel coulometric detection

This paper describes a sensitive isocratic HPLC/ECD method developed for the determination of rosmarinic acid (RA) in plant material, animal feed, and pig plasma. The plasma sample preparation only includes protein precipitation and adjustment of the pH. The applicability of the method was tested on plasma samples of pigs that were exposed to a 91-day oral intake of RA via feed enriched by aerial parts of Prunella vulgaris. The plasma was directly analyzed using the method described as well as after enzymatic hydrolysis. When no hydrolysis step was included, RA and caffeic acid (CA) were quantified in the plasma. In hydrolyzed plasma samples, several other metabolites were determined, including dihydrocaffeic, ferulic, and dihydroferulic acid. The dual-channel coulometric detection employed, as an alternative to mass spectrometry, offers good selectivity and sensitivity owing to the electrochemical properties of the phenolic constituents.     

Evaluation of isoflavone aglycon and glycoside distribution in soy plants and soybeans by fast column high-performance liquid chromatography coupled with a diode-array detector

An ultrafast HPLC/UV-vis DAD method working at 254 nm was applied for the determination of isoflavone aglycons and glycosides (genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin, and biochanin A) in roots, stems, leaves, and soy pods of soy plants and in soybeans of five varieties (Korada, Quito, Rita, OAC Erin, and OAC Vison). An Atlantis dC18 ultrafast RP chromatographic column (20 mm x 2.1 mm, 3 microm particle size) was applied for separation of the isoflavone aglycons and glycosides. A flow rate of the mobile phase (0.1% (v/v) acetic acid, pH 3.75-solvent A and methanol-solvent B) was 0.35 mL min(-1), and the column temperature was 36 degrees C. A linear gradient profile from 13 up to 22% B (v/v) from zero to 2.5 min, up to 30% B to 3.21 min, up to 35% B to 4 min, up to 40% B to 4.5 min, up to 50% B to 5.14 min, and followed by negative gradient up to 13% B to 7.71 min was used. The absolute limits of detection per sample injection (5 microL) were the highest for biochanin A (166.2 fmol) and the lowest for genistin (17.0 fmol), respectively. An accelerated solvent extraction (ASE) in combination with sonication was applied for isolation of biologically active compounds. A solid-phase extraction procedure was used to purify the extracts in the case of analysis of soy plants parts. The recoveries of 96-106% were obtained for the different concentrations of the isoflavone aglycons and glycosides and the different matrixes (overall RSDs 2-9%). The highest isoflavone concentrations were found in roots (12.5 microg g(-1) dry weight), while the amounts were about 3-1100 microg g(-1) fresh weight in different varieties of soybeans.     

The toxicity of selected acaricides against five stored product mites under laboratory assay

In laboratory tests, the toxicity of acaricides targeted against house dust mites was tested on five species of stored product mites (Acarus siro, Aleuroglyphus ovatus, Carpoglyphus lactis, Lepidoglyphus destructor, and Tyroborus lini). The formulations of benzyl-benzoate, benzyl-benzoate/permethrin/pyriproxyfen, and neem were diluted in water and applied to filter paper in an unventilated chamber. The mortality of mites was observed after 24 h of exposure to acaricide-impregnated filter paper. All of the tested acaricides were toxic to all of the mite species. There were significant differences in mortality among the species and the acaricides. Benzyl-benzoate/permethrin/pyriproxyfen was the most effective, followed by benzyl-benzoate and neem. L. destructor (LD₅₀ 0.01–0.11 μg) was the most sensitive mite species, followed by A. siro (LD₅₀ 0.04–0.12 μg), T. lini (LD₅₀ 2–21 μg), A. ovatus (LD₅₀ 3–18 μg), and C. lactis (LD₅₀ 4–64 μg). Based on the highly toxic effects of the tested acaricides against the stored product mites, the acaricides should be considered as a potential tool in the control of stored product mites, although next screening is necessary.     

Mineralization of carbon atoms of 14C-2,4-D side chain and degradation ability of bacteria in soil

The time-course of ¹⁴CO₂ formation in chernozem soil samples enriched with 1- or 2-¹⁴C-2, 4-dichlorophenoxyacetic acid (50 μg g g^?1 air-dry soil) was determined during incubation at 28°C. Except for the initial phase of decomposition, when the conversion of carboxyl carbon to ¹⁴CO₂ predominated over that of carbon in position 2, the rates of mineralization of the two carbon atoms of the side chain of the herbicide molecule exhibited no significant difference. The exponential phase of ¹⁴CO₂ evolution lasted from the 3rd to the 21st day of incubation; a semilogarithmic plot of its time dependence was strictly linear. The mineralization activity doubling time in this phase was 89.1 ± 3.6 h with 1-¹⁴CO-2, 4-D and 85.4 ± 5. l h with 2-¹⁴CO-2,4-D. An exponential decrease in mineralization activity was observed after 21 days, probably due to substrate exhaustion. The total proportion of radioactive carbon introduced into the soil in the form of 1- or 2-¹⁴CO-2,4-D and converted into ¹⁴CO₂ during 31 days of incubation was about 33%. Plate counts of bacteria increased during 35 days of incubation from 2.14 × 10⁸ to 2.8 × 10⁸ g^?1. The proportion of bacteria capable of producing ¹⁴CO₂ from the labelled herbicide increased in this period from 4.1 to 86.1%. This increase is probably directly responsible for the immediate onset of mineralization of the herbicide in soil treated previously with it or in soil inoculated with a suspension prepared from a sample previously incubated with the herbicide.     

Induction of heme oxygenase-1 by Macleaya cordata extract and its constituent sanguinarine in RAW264.7 cells

Macleaya cordata (plume poppy) is used in traditional Chinese medicine for its anti-inflammatory and antibacterial activities. In this study, we examined whether M. cordata extract and/or its major alkaloid constituents, protopine, allocryptopine, sanguinarine and chelerythrine activate the Nrf2 signalling pathway which regulates the expression of cytoprotective enzymes including heme oxygenase-1 (HO-1) and thioredoxin 1. In murine macrophage RAW264.7 cells, M. cordata extract increased both mRNA and protein levels of HO-1. Of the alkaloids examined, only sanguinarine appeared to be responsible for these effects. At the concentration of 2 μM, sanguinarine induced nuclear accumulation of Nrf2, increased the expression of Hmox1 gene encoding HO-1 and elevated HO-1 protein levels. Sanguinarine-induced Hmox1 mRNA expression was suppressed by SB203580, a pharmacologic inhibitor of p38 mitogen-activated protein kinases (p38 MAPKs). The upregulation of HO-1 in RAW264.7 cells by sanguinarine was, however, accompanied by decrease in cell viability. Nonetheless, sanguinarine at micromolar, non-cytotoxic concentrations elevated protein levels of HO-1 and thioredoxin 1 in primary cultures of human hepatocytes. We conclude that sanguinarine may, under appropriate conditions, increase the capacity of the enzymatic antioxidant defence system via activation of the p38 MAPK/Nrf2 pathway.