Determination of chromium in whole blood by DRC-ICP-MS: spectral and non-spectral interferences.

Quantification of chromium in whole blood has been performed by ICP-quadrupole MS. The spectrometer was equipped with a dynamic reaction cell (DRC) with ammonia as reaction gas. The rejection parameter q (RPq) of the DRC and the flow rate of ammonia (NH3) were optimized and set at 0.7 and 0.6 mL min(-1), respectively. Blood was diluted 1:51 (v/v) with an aqueous solution containing 0.1 mg L(-1) NH4OH, 0.1 g L(-1) EDTA, 5 mg L(-1) n-butanol, and 0.1 per thousand Triton X100. Non-spectral matrix effects observed when using the DRC were confirmed by use of vanadium. External calibration with blank and standard solutions prepared in purified water led to biased results for quality control samples. Standard addition calibration was therefore used and its validity verified. By comparing the slopes and calculating residues, it was proved that the plot obtained with standard additions and the plot obtained from blood samples of different concentrations were aligned down to 0.05 microg L(-1) after dilution.

Comparative virulence between different strains of Listeria in zebrafish (Brachydanio rerio) and mice.

Listeriosis is a disease found in most animal species but is relatively uncommon in fish. We studied the relationship between Listeria and zebrafish by injecting Brachydanio rerio intraperitoneally with different Listeria strains having pathological or food-stuff origins. We then compared these results with those obtained in Swiss mice. Experimental Listeriosis in Zebrafish differs greatly from that observed in mice. The 50% lethal dose (LD50) previously determined was much higher than that observed in mice. In fish, a good correlation exists between infection found in renal tissue, an important lymphoïd organ and that present in whole fish (p < 0.001). Infection kinetics showed that, in contrast with mice, L. monocytogenes was unable to multiply in fish. Differential blood counts showed the development of an immune response in fish. The difference in the expression of Listeria virulence between Zebrafish and mice was also seen in their reactions to different wild strains inoculate i.p. Strains belonging the innocua, ivanovii, seeligeri and welshimeri were weakly or not virulent in mice but virulent in fish. Nevertheless, as in mice, differences in virulence existed between strains of L. monocytogenes belonging to serovars 4b, 1/2a, 1/2b and 1/2c.

Regulation of De-N-Glycosylation Enzymes in Germinating Radish Seeds.

The activities of the de-N-glycosylation enzymes endo-N-acetyl- [beta]-D-glucosaminidase (ENGase; EC 3.2.1.96) and peptide-N4- (N-acetyl-[beta]-D-glucosaminyl) asparagine amidase (PNGase; EC 3.5.1.52) were monitored during germination and postgerminative development in radish (Raphanus sativus L. cv Flamboyant). The ENGase activity was detected only during postgermination, whereas the PNGase activity was present at high levels in both stages. When germination was inhibited with abscisic acid or cycloheximide, PNGase activity was detected at a basic level and ENGase activity was not detected at all. PNGase is present as an active protein in dry seeds and is apparently synthesized during seed formation. Conversely, the absence of ENGase in dry seeds suggests that its activity is dependent on the protein synthesis that occurs during and after germination. Treatment with gibberellic acid confirmed the production of both de-N-glycosylation enzymes after germination, and demonstrated a temporal delay between the production of the two enzymes during this period. Our results suggest that the two de-N-glycosylation enzymes are differentially regulated during plant development.

Copper and zinc exposure of zebrafish, Brachydanio rerio (Hamilton-Buchaman): effects in experimental Listeria infection.

To investigate the effects of heavy metals on susceptibility of fish to Listeriosis, normal zebrafish, Brachydanio rerio (Hamilton-Buchanan), were exposed to varying concentrations of zinc (0.05, 0.15, and 0.25 mg/liter) and copper (0.05, 0.10, and 0.15 mg/liter). During copper exposure, this heavy metal did not accumulate in zebrafish kidney. Unlike copper, a small amount of zinc accumulated in kidneys of fish exposed at 0.25 mg/liter. To estimate the effects of this heavy metal on listerial infection, the mortality of fish and the number of viable bacteria in fish kidney were determined at various times (1, 4, 7, and 10 days) after ip challenge with Listeria monocytogenes (strain 31386, serotype 4b). The results indicate that the number of colony-forming units in zinc-exposed fish decreased at 4, 7, and 10 days after challenge with 0.2 x LD50 of viable bacteria. In contrast, copper-exposed fish indicated both decreases and increases in the number of colony-forming units depending on the concentration of L. monocytogenes used.

Endo-N-acetyl-beta-D-glucosaminidase and peptide-N4-(N-acetyl-glucosaminyl) asparagine amidase activities during germination of Raphanus sativus.

Endo-N-acetyl-beta-D-glucosaminidase (ENGase, EC 3.2.1.96) and peptide-N4-(N-acetyl-beta-D-glucosaminyl) asparagine amidase (PNGase, EC 3.5.1.52) activities were monitored during germination and postgerminative development in Raphanus sativus. The PNGase activity was found in dry seeds and its level was constant during germination and postgermination. The ENGase activity was first detected about 18 hr after the start of imbibition (HAI) and displayed a maximum level at 36 HAI. After 36 HAI the production of both enzymes was constant until days 4-5. Both enzymes displayed substrate specificities corresponding to the potential glycoprotein substrates found in plants. They are in agreement (i) with the hypothesis that ENGase and PNGase are at the origin of the production of 'unconjugated N-glycans' and (ii) with the possibility that protein activity could be regulated by the removal of N-glycans.

Do de-N-glycosylation enzymes have an important role in plant cells?

In this review de-N-glycosylation was defined as the removal of the glycan(s) from a N-glycosylprotein, by means of enzymes acting on the di-N-acetylchitobiosyl part of the invariant pentasaccharide inner-core of N-glycosylproteins. Peptide-N4-(N-acetyl-beta-D-glucosaminyl) asparagine amidases (PNGase) and endo-N-acetyl-beta-D-glucosaminidases (ENGase) were both considered as de-N-glycosylation enzymes. A detailed description of the characterization and the function of plant PNGases and ENGases is presented, together with a brief presentation on the occurrence and the current knowledge on the function of microbial and animal enzymes. De-N-glycosylation of plant glycoproteins was proposed as a possible mechanism for the release of oligosaccharides displaying biological activities and the removal of N-glycans could also explain the regulation of protein activity. Each enzyme seems to have a specific function during germination and post-germinative development. All the arguments concur that de-N-glycosylation enzymes have an important role in plant cells and confirm that the N-glycosylation/de-N-glycosylation system should occur more commonly than presently recognized in living organisms.

[Virulence of Listeria monocytogenes in pregnant mice]. / Virulence de Listeria monocytogenes sur des souris gestantes.

Serious food-borne outbreaks of listeriosis have been reported in North America and in Europe, during the past decade. The predominant risk groups appear to be immunocompromised adults, elderly people, newborn babies and pregnant women. In order to examine the relationship between alimentation, listeriosis and pregnant females, we developed an experimental model using Swiss mice fed ad libitum during 4 days with pellets containing a high concentration of Listeria monocytogenes serovar 4b (10(9) u.f.c/g). Samples were taken from many series of pregnant mice which had been infected respectively by L. monocytogenes from 6th, 10th, 14th and 18th day of pregnancy onwards. This was compared to non infected control series. The transmission of infection from mother to progeny and contamination of surviving progeny were evaluated by Listeria numeration in liver, brain and intestines. Females infected between day 6 and day 10, and between day 10 and day 14 after fertilization, aborted or died of encephalitis. Mice contaminated between day 14 and day 18, were the least prone to experimental listeriosis. On the other hand, some mice contaminated between day 18 and day 22, i.e. at the end of their pregnancy, may develop encephalitis a few days after parturition of a healthy litter. Series contaminated between day 6 and day 10, and between day 10 and day 14 turned out to be highly sensitive to the transmission of infection from mother to young. In two other series (day 14--day 18; day 18--day 22), the young mice contained generally no Listeria. Our experimental model shows the relationship between listeriosis and alimentation. In pregnant mice, sensitivity to infection depends on their gestational status with large individual variability.