Quantification of 4-methylimidazole in class III and IV caramel colors: validation of a new method based on heart-cutting two-dimensional liquid chromatography (LC-LC)

4-Methylimidazole (4MeI) is a nitrogen compound formed during the manufacture of class III and IV caramel colors. The European Commission has limited its content to 250 ppm. Two methods were compared to perform 4MeI quantification in caramels. The first one, currently used and considered to be the reference method, consists of a hot extraction of caramel color with dichloromethane and an analysis of the acetyl derivative of the extract by gas chromatography coupled to mass spectrometry (GC-MS). The second method is based on the heart-cutting two-dimensional liquid chromatography technique (LC-LC) to directly separate 4MeI from the other components present in caramel color sample (diluted in water) in <30 min. The accuracy profile validation method and the comparison between the results obtained with the two methods show that the new and completely automated LC-LC method is usable to quantify 4MeI in caramels.     

Contamination of pastures by polycyclic aromatic hydrocarbons (PAHs) in the vicinity of a highway

To assess PAH contamination pastures, grass and soil samples have been collected from 10 m (d1), 50 m (d2), and 150 m (d3) perpendicular to a French highway (70,000 vehicles per day) and at a control site in a rural area away from nearby contaminating sources. Total PAH concentration ranges from 767 ng/g dry weight to 3989 ng/g dry weight, according to the matrix and the distance from the highway. Distance is not a significant factor for PAH deposition on grass, while in soil it has an effect between d1 and d2 or d3. The total PAH concentration in highway samples is 8 times higher than in control site samples for grass and 7 to 4 times higher for soil. Fluoranthene, pyrene, and phenanthrene are the major PAHs in grass samples at the control site and the highway, but the concentrations are about 5 times higher near the highway. In soil samples collected near the highway, the values of concentrations between all compounds are not statistically different. PAH deposition on grass is linked to the physicochemical properties of the compounds, which lead to a specific distribution of each molecule (according to their volatility and the number of aromatic rings) while no specific behavior is revealed in soil.     

Detection of polycyclic aromatic hydrocarbon levels in milk collected near potential contamination sources

Polycyclic aromatic hydrocarbons (PAHs), mainly formed by incomplete anthropogenic organic matter combustion, are ubiquitous in the environment. To assess milk PAH contamination sources, milk samples were collected from the tank milk at farms located near potential contaminating emission sources such as cementworks, steelworks, and motorways. PAH analyses were carried out by gas chromatography coupled to mass spectrometry. Eight PAHs were identified in milk: naphthalene, acenaphthylene, acenaphthene, fluorene, anthracene, fluoranthene, pyrene, and benzo[a]anthracene. For all potential contaminating sources, these eight PAHs were detected with similar profiles and at low concentrations except for fluorene and naphthalene, for which source-molecule interaction is pointed out.     

The Pho regulon and the pathogenesis of Escherichia coli

During the course of infection, bacteria must coordinately regulate gene expression in response to environmental stimuli. The phosphate (Pho) regulon is controlled by the two component-regulatory system PhoBR. PhoBR is activated during starvation and regulates genes involved in phosphate homeostasis. Several studies have highlighted the importance of the Pho regulon in bacterial pathogenesis, showing how induction of PhoBR, in addition to regulating genes participating in phosphate metabolism, leads to modulation of many cellular processes. The pleiotropic effects of Pho regulon activation include attenuated virulence and alteration of many virulence traits, including adhesion to host cells and resistance to cationic antimicrobial peptides, acidity and oxidative stresses. This review provides an overview of the relationship between the Pho regulon and virulence in Escherichia coli and illustrates that, in addition to regulating phosphate homeostasis, the Pho regulon plays a key role in regulating stress responses and virulence.     

Zur Frage der Gefährdung von Weidevich bei Anwendung der Flächenbehandlung gegen Feldmäuse (Microtus arvalis PALLAS) auf Grünland

Zusammenfassung Die in eigenen Versuchen ermittelten Minimum - Dosierungen von Toxaphen und Endrin bei der Flächenbehandlung gegen Feldmäuse (Microtus arvalis PALLAS) werden an Hand der von anderer Seite veröffentlichten Untersuchungen zur Toxizität der Wirkstoffe für Weidevieh diskutiert. Die vorliegenden Versuche mit Futter, das unmittelbar nach der Applikation der Wirkstoffe verabreicht wurde, lassen jedoch keine Rückschlüsse auf die Verhältnisse bei der Feldmaus-Flächenbegiftung zu. Denn bei der Durchführung dieses Verfahrens während der Vegetationsruhe werden die Wirkstoffe in der zwangsläufig gegebenen Wartezeit bis zur Wiederbeweidung abgebaut, so daß es kaum zur Bildung von bedenklichen Rückstandsmengen in Fleisch, Fett und Milch der Weidetiere kommen kann. Eine einmalige Flächenbehandlung gegen Feldmäuse auf Grünland mit den jetzt erreichten Minimal-Dosierungen, die eine weitere Annäherung an den insektiziden Bereich bedeuten, ist augenscheinlich in toxikologischer Hinsicht nicht anders zu bewerten als z. B. eine zweimalige Behandlung von Kulturpflanzen mit insektiziden Konzentrationen.     

Effects of a standardized anesthetic protocol on hematologic variables in healthy cats

This study evaluated the effects of an anesthetic protocol using intravenous ketamine and midazolam, and intramuscular buprenorphine on hematologic variables in cats. Twelve healthy adult cats had blood collected for a complete blood count before and after the induction of anesthesia. There were significant decreases in red blood cell counts, hemoglobin concentrations and hematocrits after the induction of anesthesia. On average, red blood cell counts and hematocrits decreased by 25%, and hemoglobin concentrations decreased by 24%. Based on hematocrit, 3/12 samples (25%) taken while the cats were anesthetized would have been interpreted as belonging to anemic patients while none of the cats would have been considered anemic before anesthesia. This study suggests that a complete blood count performed on blood taken under anesthesia with this anesthetic protocol should be interpreted cautiously in order to not make a false diagnosis of anemia.     

Major biocontrol of plant tumors targets tRNA synthetase

Crops can be devastated by pathogenic strains of Agrobacterium tumefaciens that cause crown gall tumors. This devastation can be prevented by the nonpathogenic biocontrol agent A. radiobacter K84, which prevents disease by production of the "Trojan horse" toxin agrocin 84, which is specifically imported into tumorgenic A. tumefaciens strains to cause cell death. We demonstrate that this biocontrol agent targets A. tumefaciens leucyl-tRNA synthetase (LeuRS), an essential enzyme for cell viability, while the agent itself survives by having a second, self-protective copy of the synthetase. In principle, this strategy from nature could be applied to other crop diseases by direct intervention.     

An antifungal agent inhibits an aminoacyl-tRNA synthetase by trapping tRNA in the editing site

Aminoacyl-transfer RNA (tRNA) synthetases, which catalyze the attachment of the correct amino acid to its corresponding tRNA during translation of the genetic code, are proven antimicrobial drug targets. We show that the broad-spectrum antifungal 5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole (AN2690), in development for the treatment of onychomycosis, inhibits yeast cytoplasmic leucyl-tRNA synthetase by formation of a stable tRNA(Leu)-AN2690 adduct in the editing site of the enzyme. Adduct formation is mediated through the boron atom of AN2690 and the 2'- and 3'-oxygen atoms of tRNA's3'-terminal adenosine. The trapping of enzyme-bound tRNA(Leu) in the editing site prevents catalytic turnover, thus inhibiting synthesis of leucyl-tRNA(Leu) and consequentially blocking protein synthesis. This result establishes the editing site as a bona fide target for aminoacyl-tRNA synthetase inhibitors.