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Biomedical subjects

S Aubert

Publications and source records attributed to S Aubert.

At least 37 records · Page 2Linked to original sources

The value of rRNA gene restriction site polymorphism analysis for delineating taxa in the genus Staphylococcus.

A total of 101 staphylococcal strains were ribotyped using EcoRI and HindIII as restriction enzymes and plasmid pBA2 as the rDNA probe. Isolates from 10 newly described staphylococcal taxa were among those examined. All the ribotypes were added to our database, Staph DB, which now contains the sizes of the bands of 135 EcoRI and 120 HindIII ribotypes from 408 strains belonging to 42 staphylococcal taxa. The relatedness of ribotypes was evaluated by using the Dice coefficient. The ribotypes, and thus the strains, were clustered by the unweighted pair group method with averages (UPGMA). Separation into clusters correlated well with the delineation of the staphylococcal species but not with that of the different subspecies. No discrimination was possible between Staphylococcus vitulinus and Staphylococcus pulvereri. Ecovar-specific groups were evident within Staphylococcus intermedius and Staphylococcus hyicus. The data increase the usefulness of rRNA gene restriction site polymorphism analysis for staphylococcal taxonomy.

Animals↗

Metabolism of methanol in plant cells. Carbon-13 nuclear magnetic resonance studies.

Using (13)C-NMR, we demonstrate that [(13)C]methanol readily entered sycamore (Acer pseudoplatanus L.) cells to be slowly metabolized to [3-(13)C]serine, [(13)CH(3)]methionine, and [(13)CH(3)]phosphatidylcholine. We conclude that the assimilation of [(13)C]methanol occurs through the formation of (13)CH(3)H(4)Pte-glutamate (Glu)(n) and S-adenosyl-methionine, because feeding plant cells with [3-(13)CH(3)]serine, the direct precursor of (13)CH(2)H(4)Pte-Glu(n), can perfectly mimic [(13)CH(3)]methanol for folate-mediated single-carbon metabolism. On the other hand, the metabolism of [(13)C]methanol in plant cells revealed assimilation of label into a new cellular product that was identified as [(13)CH(3)]methyl-beta-D-glucopyranoside. The de novo synthesis of methyl-beta-D-glucopyranoside induced by methanol did not require the formation of (13)CH(3)H(4)Pte-Glu(n) and was very likely catalyzed by a "transglycosylation" process.

Amino Acids↗

[Intubation using a submental approach].

Authors report a case of sub-mental approach for endotracheal intubation in maxillo-facial surgery. This unusual technique was simple to perform and remained uncomplicated.

Adult↗

Workplace monitoring of hydrogen peroxide using titanyl-coated sorbents.

The current methods of sampling hydrogen peroxide, based on bubbling in acid solutions (titanium tetrachloride or titanium oxysulfate), are unsuitable for personal sampling. An alternative medium has been developed: silica gel coated with titanium oxysulfate. Sample tubes of this new sampling medium are suitable for personal sampling. The sampling performance is the same as that of the methods based on bubbling, but the tubes must be desorbed as soon as possible, even if the spectrophotometric analysis can be postponed. Special emphasis has been placed on the possible interference by peracetic acid, which is currently used in conjunction with hydrogen peroxide in the food industry.

Absorption↗

Glycine and serine catabolism in non-photosynthetic higher plant cells: their role in C1 metabolism.

Glycine and serine are two interconvertible amino acids that play an important role in C1 metabolism. Using 13C NMR and various 13C-labelled substrates, we studied the catabolism of each of these amino acids in non-photosynthetic sycamore cambial cells. On one hand, we observed a rapid glycine catabolism that involved glycine oxidation by the mitochondrial glycine decarboxylase (GDC) system. The methylenetetra- hydrofolate (CH2-THF) produced during this reaction did not equilibrate with the overall CH2-THF pool, but was almost totally recycled by the mitochondrial serine hydroxymethyltransferase (SHMT) for the synthesis of one serine from a second molecule of glycine. Glycine, in contrast to serine, was a poor source of C1 units for the synthesis of methionine. On the other hand, catabolism of serine was about three times lower than catabolism of glycine. Part of this catabolism presumably involved the glycolytic pathway. However, the largest part (about two-thirds) involved serine-to-glycine conversion by cytosolic SHMT, then glycine oxidation by GDC. The availability of cytosolic THF for the initial SHMT reaction is possibly the limiting factor of this catabolic pathway. These data support the view that serine catabolism in plants is essentially connected to C1 metabolism. The glycine formed during this process is rapidly oxidized by the mitochondrial GDC-SHMT enzymatic system, which is therefore required in all plant tissues.

Journal Article↗

Transport, Compartmentation, and Metabolism of Homoserine in Higher Plant Cells. Carbon-13- and phosphorus-31-nuclear magnetic resonance studies Carbon-13- and Phosphorus-31-Nuclear Magnetic Resonance Studies

The transport, compartmentation, and metabolism of homoserine was characterized in two strains of meristematic higher plant cells, the dicotyledonous sycamore (Acer pseudoplatanus) and the monocotyledonous weed Echinochloa colonum. Homoserine is an intermediate in the synthesis of the aspartate-derived amino acids methionine, threonine (Thr), and isoleucine. Using 13C-nuclear magnetic resonance, we showed that homoserine actively entered the cells via a high-affinity proton-symport carrier (Km approximately 50-60 mum) at the maximum rate of 8 +/- 0.5 mumol h-1 g-1 cell wet weight, and in competition with serine or Thr. We could visualize the compartmentation of homoserine, and observed that it accumulated at a concentration 4 to 5 times higher in the cytoplasm than in the large vacuolar compartment. 31P-nuclear magnetic resonance permitted us to analyze the phosphorylation of homoserine. When sycamore cells were incubated with 100 mum homoserine, phosphohomoserine steadily accumulated in the cytoplasmic compartment over 24 h at the constant rate of 0.7 mumol h-1 g-1 cell wet weight, indicating that homoserine kinase was not inhibited in vivo by its product, phosphohomoserine. The rate of metabolism of phosphohomoserine was much lower (0.06 mumol h-1 g-1 cell wet weight) and essentially sustained Thr accumulation. Similarly, homoserine was actively incorporated by E. colonum cells. However, in contrast to what was seen in sycamore cells, large accumulations of Thr were observed, whereas the intracellular concentration of homoserine remained low, and phosphohomoserine did not accumulate. These differences with sycamore cells were attributed to the presence of a higher Thr synthase activity in this strain of monocot cells.

Journal Article↗

Analysis of two Staphylococcus epidermidis plasmids coding for resistance to streptogramin A.

The two Staphylococcus epidermidis plasmids pIP1629 (7.5 kb) and pIP1630 (14.4 kb) contain the vga gene conferring resistance to streptogramin A. All the sequences of pIP1629, except two of the four 22-nt iterons preceding the replication gene, were found in pIP1630. The additional 6.9-kb fragment of pIP1630 is similar to the mobilizable S. epidermidis plasmid pSK639, carrying the dfrA-thyE-orf140 operon and thought to replicate by an iteron controlled theta-type replication mechanism. The replication-mobilization elements of pIP1629 and pSK639 are very similar despite having been isolated in France and in Australia, respectively, showing that they are geographically widely dispersed in S. epidermidis. The gene thyE encoding thymidylate synthetase carried by pSK639 is not present in pIP1630. pIP1630 probably arose by the recombination of two homologous plasmids carrying distinct resistance determinants.

Australia↗

Exposure to chloramines in a green salad processing plant.

Acute eye and upper respiratory irritation was reported at an industrial facility that processes green salads in water containing hypochlorite. The irritant agents were chloramines resulting from the reaction of hypochlorite and nitrogen compounds coming from the sap proteins released when the vegetables were cut. Specific sampling and analytical methods were developed in order to assess the workers' exposure. Monochloramine, dichloramine and nitrogen trichloride were found in the atmosphere, which can explain the irritation phenomena of which the workers complained. In the washing room (where salads are processed), the total chloramine concentration ranged from 0.4 to 16 mg.m-3, depending on the day and the location of the area sampling points. The exposure of workers determined by personal sampling ranged from 0.2 to 5 mg.m-3. In a previous study, with swimming pool instructors exposed to the same pollutants, irritation phenomena generally appeared at chloramine values of around 0.5 mg.m-3.

Chloramines↗

Multiplicity of the genes and plasmids conferring resistance to pristinamycin in staphylococci selected in an Algerian hospital.

In an Algerian hospital where pristinamycin (Pt) was extensively used for the treatment of chronic osteomyelitis and for prophylaxis in bone surgery, the prevalence of pristinamycin-resistant (PtR) staphylococci during a five-month period (20%) was higher than that among staphylococci isolated elsewhere in Algeria (4.5%). Analysis of 13 PtR staphylococci isolated in this hospital revealed a diversity of plasmids and genes conferring resistance to Pt and to related antibiotics. Most of the PtR staphylococci were unrelated: they belonged to either different taxa or types. Nevertheless, some of the unrelated staphylococci harboured structurally related plasmids carrying streptogramin resistance genes. Thus, these plasmids may have contributed to the dispersion of these genes.

Algeria↗

Mapping the regions carrying the three contiguous antibiotic resistance genes aadE, sat4, and aphA-3 in the genomes of staphylococci.

Tn5405 (12 kb) is a staphylococcal composite transposon delimited by two inverted copies of IS1182, one of which contains IS1181. The internal part of this transposon carries three antibiotic resistance genes, aphA-3, aadE, and sat4, and three open reading frames (ORFs), orfx, orfy, and orfz, of unknown function. The dispersion of Tn5405 and the genes and ORFs included in this transposon were investigated in 50 epidemiologically unrelated staphylococci carrying aphA-3. Twenty-three maps, distinguishable by the presence or absence of the investigated genes or ORFs and/or by the sizes of the restriction fragments carrying them, were identified. Four isolates carried Tn5405, and 15 other isolates contained a Tn5405-related element. IS1182 was not detected in the aphA-3 regions mapped in 31 isolates which carried the following combinations: orfx, orfy, aadE, sat4, and aphA-3 +/- orfz; orfy, aadE, sat4, and aphA-3 +/- orfz; and aadE, sat4, aphA-3, and orfz. In all isolates, the genes and ORFs investigated were in relative positions similar to those in Tn5405. Thus, the internal part of Tn5405 appeared to be partially conserved with the maintenance, in all of the isolates, of at least the three antibiotic resistance genes.

DNA, Bacterial↗

Contribution of a typing method based on IS256 probing of SmaI-digested cellular DNA to discrimination of European phage type 77 methicillin-resistant Staphylococcus aureus strains.

The incidence of infections with phage type 77 methicillin-resistant Staphylococcus aureus (MRSA) strains increased in France in 1987. These strains are widespread in numerous European hospitals. The SmaI restriction profiles of total DNA extracted from 74 phage type 77 MRSA strains isolated from 1987 to 1994 in 10 hospitals in eight European cities (in France, Belgium, and Spain) were analyzed. Hybridization with a probe containing a 468-bp DNA fragment from within the transposase gene of the insertion sequence IS256 was also examined. Forty-three SmaI profiles were detected. Twenty major genotypes were identified, and each genotype contained strains with the same profile or profiles which differed by no more than three bands. Strains isolated in different countries and at several-year intervals were often grouped within the same genotype. A larger number of genotypes could be discriminated by analysis of the patterns of hybridization with the IS256 probe. SmaI restriction fragments with the same apparent electrophoretic mobility could, in some cases, be distinguished by the presence or the absence of nucleotide sequences hybridizing with IS256. The strains that grouped within the same genotype after hybridization with IS256 were mostly those isolated in the same hospital and at less than 12-month intervals. Consequently, the IS256 probe that we used improved restriction profile analysis for discrimination between the intrahospital, outbreak-related phage type 77 MRSA strains and the endemic strains disseminated in various cities and countries.

Bacterial Typing Techniques↗

Arrest of mitochondrial biogenesis in copper-treated sycamore cells.

Sycamore suspension cells (Acer pseudoplatanus L.) were grown in the presence of sublethal concentrations of copper (50 microM). During the first 5-6 days of treatment, growth was not affected, but cell respiration (coupled and uncoupled) declined to approximately 60% of its normal value. This decline of respiration was attributed to a progressive diminution of the number of mitochondria in copper-treated cells, based on the demonstration of the concomitant decline of (1) cardiolipin (diphosphatidylglycerol) and cytochrome aa3 (cytochrome oxidase), two specific markers of mitochondrial inner membrane, and (2) fumarase activity, a specific marker of mitochondrial matrix space. In addition, the mitochondria extracted from copper-treated cells presented the same properties as those from control cells, concerning substrate oxidation, cardiolipin and cytochrome aa3 contents, and fumarase activity. These results strongly suggest that copper triggered an arrest of mitochondrial biogenesis, which preceded cell division arrest.

Cardiolipins↗

Induction of beta-methylcrotonyl-coenzyme A carboxylase in higher plant cells during carbohydrate starvation: evidence for a role of MCCase in leucine catabolism.

Induction of beta-methylcrotonyl-coenzyme A carboxylase (MCCase) activity was observed during carbohydrate starvation in sycamore cells. In mitochondria isolated from starved cells, we noticed a marked accumulation of the biotinylated subunit of MCCase, of which the apparent molecular weight of 74000 was similar to that of the polypeptide from mitochondria of potato tubers. Our results provide evidence for a role of MCCase in the catabolic pathway of leucine, a branched-chain amino acid which transiently accumulates in carbon-starved cells in relation to a massive breakdown of proteins. Furthermore, when control sycamore cells were incubated in the presence of exogenous leucine, this amino acid accumulated in the cells and no induction or accumulation of MCCase was observed, indicating that leucine is not responsible for the induction of its catabolic machinery. Finally, MCCase is proposed as a new biochemical marker of the autophagic process triggered by carbohydrate starvation.

Carbon-Carbon Ligases↗

Ultrastructural and biochemical characterization of autophagy in higher plant cells subjected to carbon deprivation: control by the supply of mitochondria with respiratory substrates.

Autophagy triggered by carbohydrate starvation was characterized at both biochemical and structural levels, with the aim to identify reliable and easily detectable marker(s) and to investigate the factors controlling this process. Incubation of suspension cells in sucrose-free culture medium triggered a marked degradation of the membrane polar lipids, including phospholipids and galactolipids. In contrast, the total amounts of sterols, which are mainly associated with plasmalemma and tonoplast membranes, remained constant. In particular, phosphatidylcholine decreased, whereas phosphodiesters including glycerylphosphorylcholine transiently increased, and phosphorylcholine (P-Cho) steadily accumulated. P-Cho exhibits a remarkable metabolic inertness and therefore can be used as a reliable biochemical marker reflecting the extent of plant cell autophagy. Indeed, whenever P-Cho accumulated, a massive regression of cytoplasm was noticed using EM. Double membrane-bounded vacuoles were formed in the peripheral cytoplasm during sucrose starvation and were eventually expelled into the central vacuole, which increased in volume and squeezed the thin layer of cytoplasm spared by autophagy. The biochemical marker P-Cho was used to investigate the factors controlling autophagy. P-Cho did not accumulate when sucrose was replaced by glycerol or by pyruvate as carbon sources. Both compounds entered the cells and sustained normal rates of respiration. No recycling back to the hexose phosphates was observed, and cells were rapidly depleted in sugars and hexose phosphates, without any sign of autophagy. On the contrary, when pyruvate (or glycerol) was removed from the culture medium, P-Cho accumulated without a lag phase, in correlation with the formation of autophagic vacuoles. These results strongly suggest that the supply of mitochondria with respiratory substrates, and not the decrease of sucrose and hexose phosphates, controls the induction of autophagy in plant cells starved in carbohydrates.

Autophagy↗

Distribution of genes encoding resistance to streptogramin A and related compounds among staphylococci resistant to these antibiotics.

The levels of resistance to pristinamycin (Pt) and to its major constituents, pristinamycin IIA and IB (PIIA and PIB, respectively; classified as streptogramins A and B, respectively) were determined for 126 staphylococcal isolates. The results suggest tentative susceptibility breakpoints of < or = 2, < or = 8, and < or = 0.5 microgram/ml for PIIA, PIB, and Pt, respectively. Fifty-six isolates that were inhibited by > or = 4 micrograms of PIIA per ml were investigated for the presence of staphylococcal genes encoding resistance to PIIA (vga, vat, and vatB) and PIB (vgb). None of these genes was found in the 4 isolates inhibited by 4 micrograms of PIIA per ml or in 4 of the other 52 isolates tested. The remaining 48 isolates harbored plasmids carrying vatB and vga or combinations of genes (vga-vat-vgb or vga-vat). The absence of any known PIIA resistance gene from the four Staphylococcus aureus isolates inhibited by > or = 8 micrograms of PIIA per ml suggests that there is at least one PIIA resistance mechanism in staphylococci that has not yet been characterized.

Anti-Bacterial Agents↗

Multiple effects of glycerol on plant cell metabolism. Phosphorus-31 nuclear magnetic resonance studies.

The effects of glycerol on plant cell metabolism were studied with sycamore (Acer pseudoplatanus L.) cells using 31P nuclear magnetic resonance spectroscopy. After a long period of sucrose starvation, the addition of 50 mM glycerol to the medium did not restore the original glucose-6-P pool and led to a rapid accumulation of sn-glycerol-3-P in the cytoplasmic compartment. The synthesis of sn-glycerol-3-P was rapid and occurred first at the expense of cytoplasmic P(i). Accumulated sn-glycerol-3-P competitively inhibited glucose-6-phosphate isomerase activity when fructose-6-P was the varied substrate. Such a situation prevented the rapid recycling of triose phosphates back to hexose phosphates and led to an arrest of the functioning of the cytosolic and plastidial pentose phosphate pathways. Under these conditions, the flow of carbon to drive cell respiration derived almost exclusively from glycerol, and this polyalcohol was not used as a source of carbon skeletons for biosynthesis. Glycerol also induced the accumulation of O-phosphohomoserine in the cytoplasmic compartment as long as the cell culture medium contained sucrose. Finally glycerol added to sucrose-starved cells stopped the accumulation of phosphocholine (Roby, C., Martin J.-B., Bligny, R., and Douce, R. (1987) J. Biol. Chem. 262, 5000-5007) and prevented a further decline in the uncoupled rate of O2 consumption by the cells (Journet, E. P., Bligny, R., and Douce, R. (1986) J. Biol. Chem. 261, 3193-3199). These last observations strongly suggest that glycerol prevented the triggering of autophagy induced by sucrose starvation in sycamore cells.

Adenosine Triphosphate↗

Antimicrobial activity of shredded carrot extracts on food-borne bacteria and yeast.

Purified ethanolic extracts of peeled and shredded carrots showed an antimicrobial effect against a range of food-borne micro-organisms. The minimum inhibitory concentration, expressed as mg ml-1 dried carrot material used for the extraction were: Leuconostoc mesenteroides, 27; Listeria monocytogenes, > 27 < 55; Staphylococcus aureus, > 27 < 55; Pseudomonas fluorescens, > 55 < 110; Candida lambica, > 55 < 110; Escherichia coli, > 110 < 220. The antimicrobial activity was not linked to phenolic compounds but was presumably due to apolar components. Free saturated fatty acid (dodecanoic acid) and methyl esters of saturated fatty acids (of dodecanoic and pentadecanoic acids) were identified in purified active extracts of carrots by gas chromatography coupled to mass spectrometry and could be responsible for the antimicrobial activity. This effect did not seem to play a role in the resistance of shredded carrots to microbial spoilage, although the antimicrobial activity was present in fresh carrots at concentrations sufficient to inhibit spoilage bacteria.

Anti-Bacterial Agents↗