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Biomedical subjects

J Etienne

Publications and source records attributed to J Etienne.

At least 19 recordsLinked to original sources

Sequence of rat lipoprotein lipase-encoding cDNA.

A rat lipoprotein lipase (LPL)-encoding cDNA (LPL) has been entirely sequenced and compared to the sequences of all the LPL cDNAs reported in other species. As expected, high homology was found between the coding exons. The putative catalytic triad, Ser132, Asp156, His241, according to human numbering, is conserved in rat. As is the case in mouse, an Asn444 present in human LPL is also missing. The major divergences between human, mouse and rat LPLs were observed in the untranslated exon 10, where (i) the rat cDNA exhibits a 157-bp insertion and an 81-bp deletion relative to human; (ii) neither the B1 repeat nor the homopurine stretch reported in mouse can be recognized, and (iii) the rat cDNA displays several A+T-rich stretches.

Amino Acid Sequence

Cadmium-resistance plasmid in Staphylococcus lugdunensis.

Plasmids that confer resistance to cadmium (MIC > or = 125 microM), were found in 18 out of 30 independent Staphylococcus lugdunensis strains from clinical specimens. Variants that were cured of their plasmid were cadmium sensitive. Restriction endonuclease sites of a 3.2-kb cadmium-resistance plasmid of S. lugdunensis, designated pLUG10, were similar to those of pOX6, a S. aureus cadmium-resistance plasmid containing the cadB gene. Southern-blot hybridisation was performed with a probe intragenic to cadB. Hybridisation was found with the cadB probe in the cadmium-resistant S. lugdunensis isolates to the 2.9-, 3.2- and 3.7-kb plasmids. These findings suggest that cadmium-resistance in some S. lugdunensis strains is due to a gene sharing homology with the cadB gene of S. aureus.

Cadmium

Comparison of coagulase-negative staphylococci by pulsed-field electrophoresis.

Five pathogenic strains each of Staphylococcus epidermidis, S. haemolyticus, S. lugdunensis and S. schleiferi were analysed by conventional electrophoresis and field inversion gel electrophoresis. For these coagulase-negative staphylococci, the restriction endonuclease SmaI emerged as the most suitable enzyme for pulsed-field electrophoresis by providing an adequate number of clearly separated DNA fragments. Field inversion gel electrophoresis confirmed the differences among strains already discriminated by conventional electrophoresis, and furthermore, differentiated strains which had previously appeared identical. Among the species that were studied, S. epidermidis showed great genomic diversity with a few common bands. On the contrary, S. haemolyticus, S. lugdunensis and S. schleiferi showed less diversity. Although these minor variations may be epidemiologically significant, this question has to be investigated on a larger number of strains.

Coagulase

Treatment of streptococcal endocarditis with a single daily dose of ceftriaxone sodium for 4 weeks. Efficacy and outpatient treatment feasibility.

UNLABELLED: OBJECTIVE--To evaluate the efficacy and safety of ceftriaxone sodium in the treatment of streptococcal endocarditis. DESIGN--An open, multicenter, noncomparative study with a follow-up of patients for 4 months to 5 years. SETTING--Internal medicine wards and outpatient clinics of hospitals of various sizes in three European countries. PATIENTS--Fifty-nine patients with defined criteria for streptococcal endocarditis. INTERVENTION--Ceftriaxone sodium administered at a once-daily dose of 2 g for 4 weeks. MAIN OUTCOME MEASURES--Clinical outcome and microbiological cure rate. RESULTS--Among the 59 patients, 55 completed the treatment and were followed up for 4 months to 5 years. No patients showed evidence of relapse. Treatment was completely uneventful in 42 patients (71%). A cardiac valve was replaced in four patients (7%) receiving antimicrobial therapy and in six patients (10%) who had completed antimicrobial therapy. One of the 10 valves taken for culture at surgery was positive, but only for microorganisms that were different from the microorganism isolated before the treatment. The treatment had to be interrupted in four patients because of drug allergy. Other side effects were mild except for two cases of reversible neutropenia. The treatment was easy to administer: 27 patients (46%) had no permanent intravenous catheter at any time, seven patients (12%) had such a catheter for less than 4 days. Twenty-three patients (39%) were discharged from the hospital less than 2 weeks after admission. CONCLUSIONS: --Ceftriaxone sodium administered at a once-daily dose of 2 g appears to be an effective and safe treatment of streptococcal endocarditis. In hospitals, this agent may be more convenient to administer than penicillin G with or without aminoglycosides. Some patients may even be treated as outpatients.

Adult

An attempt to reconstruct the lithotriptor shock wave pulse in kidney: possible temperature effects.

Based on measurements carried out in water in two lithotriptor systems, the authors have made an attempt to reconstruct numerically amplitudes and shapes of shock wave pulses penetrating into kidney which differ from those in water. The difference between these pulses and those observed in water was analyzed and was also demonstrated experimentally. The amplitude and the steepness of the reconstructed pulse front were shown to be much lower than in water depending on the distance of the kidney stone from the patient's body surface. For a distance equal to 4 cm, the shock wave pulse amplitude of 40 MPa in water was estimated to decrease in the kidney by a factor of about two and the steepness of the positive shock pulse front to decrease several times. The analysis was carried out by considering the possible changes of absorption and attenuation in tissues which increase in an unknown way with the wave amplitude. It was shown that the temperature elevation caused by the increase of nonlinear high amplitude absorption is limited due to a corresponding increase in attenuation of the shock wave penetrating soft tissues. The temperature elevation was estimated on the basis of this work to be at most 1.8 times that one estimated in the case of two considered lithotripsy systems when assuming small amplitude absorption and attenuation coefficients.

Animals

Lipoprotein lipase in heart and myocytes: characteristics with intralipid as substrate.

1. Intralipid is a suitable substrate for measuring lipoprotein lipase activity in the presence of other triacylglycerol lipases in heart and myocytes. 2. Triacylglycerol lipase activity in heart and myocytes was increased 10-fold in the presence of serum at pH 7.4 and 8.1. The serum-stimulated activity in myocytes was 95% inhibited by saturating concentrations of antiserum to lipoprotein lipase. 3. Both heparin-releasable and non-releasable lipoprotein lipase fractions had similar Km values for Intralipid and a similar pattern of inhibition by high density lipoprotein but different responses to heparin. 4. Isoproterenol did not alter lipoprotein lipase activity in cardiac myocytes.

Animals

Chlamydial endocarditis: a report on ten cases.

Over the period 1983-1990, 10 cases of infective native-valve endocarditis as a result of Chlamydia were seen. All patients were men, with a mean age of 42 years, and none had a history of exposure to Chlamydia psittaci. Symptoms, such as weight loss and anorexia, with fever in eight cases, had persisted for at least 2 months before admission. Haemodynamic failure was present in seven patients, and neurological signs in four. The aortic valve was involved in seven cases, the mitral valve in one and both valves in two. Vegetations, often fingerlike, were observed by echocardiography in nine cases. All patients required valve replacement, and three died in the year following diagnosis. Blood cultures were consistently negative in all cases, and no antibiotics had been given before admission. Significant titres of complement fixing anti-chlamydial antibodies were present in six cases, and micro-immunofluorescence using the three chlamydial species showed cross-reacting antibodies in all nine cases studied, with transient IgM positivity in six cases. Histologically, the leaflets were fibrosed and infiltrated by large macrophages containing dark red granules after Machiavello staining. These granules were specifically stained by immunofluorescence using monoclonal antibody to Chlamydia common antigen, but not by that specific for C. pneumoniae. No organisms were seen after Gram staining, and no positive chlamydial immunofluorescence was seen on sections of valves from patients with staphylococcal or streptococcal endocarditis.

Adult

Induction of a putative laminin-binding protein of Streptococcus gordonii in human infective endocarditis.

There is evidence to suggest that the virulence of Streptococcus strains in infective endocarditis might be due to the expression of binding sites for the extracellular matrix proteins of damaged valves. In this communication, we draw attention to one laminin-binding protein from a strain of Streptococcus gordonii isolated from a patient with human endocarditis. This 145-kDa protein was found on the cell wall of the bacterium. The level of expression of this binding protein might be regulated by the presence of extracellular matrix proteins: the protein was lacking after in vitro selection of laminin, collagen I, and fibronectin nonbinding variants, and it was recovered after growth of the variants when laminin or collagen I was added to the growth medium. It was also missing after 10 subcultures in minimal medium, indicating some positive control. Furthermore, the 145-kDa protein was recognized as a major antigen by sera from patients treated for streptococcal infective endocarditis, while sera from patients with valvulopathies gave only slight recognition, suggesting an increase of the expression of this protein during infective endocarditis. It was also shown that the 145-kDa protein carried a collagen I-like determinant detected with anti-human collagen I antibodies.

Collagen

Description and evaluation of the semiautomated 4-hour rapid ID 32 Strep method for identification of streptococci and members of related genera.

The rapid ID 32 Strep system (bioMérieux, La Balme les Grottes, France) is a new system which allows the identification in 4 h of most streptococci and members of related genera encountered in medical and veterinary bacteriology. Four hundred thirty-three isolates first identified by conventional methods and belonging to the genera Streptococcus, Lactococcus, Enterococcus, Aerococcus, Gemella, Leuconostoc, Erysipelothrix, Gardnerella, and Listeria were tested. Overall, rapid ID 32 Strep correctly identified 413 (95.3%) of the strains, with 109 (25.1%) requiring extra tests for complete identification. Sixteen strains (3.7%) were not identified, and 4 (1.0%) were misidentified. The rapid ID 32 Strep system is a suitable alternative for rapid identification of members of the genus Streptococcus and of related genera.

Bacterial Typing Techniques

[Genes of the lipase family: comparison of nucleic and proteinic sequences].

Vertebrates' plasmatic apolipoproteins and a few number of lipases in their metabolism present sequence homologies. They are grouped in genes families. The four exons apolipoproteins gene family includes nine human genes: the divergence rate of their sequences allows to place the first ancestral gene very high in the phylogenetic tree of the evolution. However, a more recent duplication of apolipoprotein C-I gene dating from 40 millions years, may be a phylogenetic marker for the radiation of Monkeys. Pancreatic lipase and isoforms, lipoprotein-lipase and hepatic triacylglycerol-lipase form by their homologies a "superfamily" of genes, which also includes yolk proteins of Dipterians eggs. Sequence homologies of PL, LPL and HL are analysed and compared with multiple alignments of amino-acids and nucleotides on spreadsheets. From these comparisons we may characterize four classes of phylogenetic markers: 1) repetitive DNA sequence (Alu, B1, PRE-1) appeared during Mammals evolution, 2) short insertions or deletions (within N-terminal domain) and a gene conversion in guinea-pig lineage, 3) a progressive reduction of intron number during the lipases evolution, 4) several duplications of genes which have produced the five genes of this superfamily currently known in the human genome.

Amino Acid Sequence

[Lipoprotein lipase deficiencies].

Lipoprotein lipase (LPL) is an enzyme which plays a major role in the metabolism of circulating triglyceride-rich lipoproteins. It hydrolyzes chylomicron and VLDL triglycerides, thereby delivering fatty acids to tissues for storage or oxidation. In order to gain insight into the molecular basis of LPL deficiency, the structure of the LPL gene (ten exons and nine introns spanning about 30 kb) is first set out in relation to the different domains of the LPL protein. There is a high sequence homology between the aminoacids of LPL and of other lipases, such as hepatic triglyceride lipase (HL) and pancreatic lipase (PL). The PL catalytic triad Ser132, Asp156, His241 is also present in LPL. Absence of LPL activity can result from absence of LPL protein synthesis (Brunzell class I), or from the synthesis of an LPL protein devoid of enzymatic activity consequently to a mutation (class II). LPL can also be unable to bind to endothelial cells--a defect combined with deficient enzymatic activity--(class III). Among the known mutations of the LPL gene (such as nonsense, frameshift, abnormality in intron-exon junction, deletion, duplication) resulting in pathological cases, the most frequent are punctual mutations located mainly in exons 4, 5 and 6, leading to the substitution of an aminoacid for another in essential domains of LPL. The combined deficiency LPL + HL has also been described. The study of the abnormalities of the LPL gene, known only since the years 1990-1991, allows not only to better understand the pathology of LPL deficiencies, but also to point out which aminoacids play a major role in LPL activity.

DNA Mutational Analysis

[Infectious agents associated with exacerbations of chronic obstructive bronchopneumopathies and asthma attacks].

Infections of the respiratory airways are frequently responsible for exacerbations of chronic obstructive pulmonary disease (COPD) and attacks of asthma. However, the causal infectious agents in practice are rarely precisely identified. We have undertaken a prospective study with the aim of researching into the bacteria and viruses associated with these exacerbations. Forty-seven patients who were in hospital between 1987 and 1989 for attacks of asthma (13 episodes) or exacerbations of COPD (35 episodes) were included in this study. The microbiological analysis consisted of: 1) the bacteriology of expectorated material or the products aspirated by fibroscopy with direct examination, quantitative cytology and culture; 2) samples taken from the nasal airways to identify and isolate pneumotropic viruses and mycoplasma; 3) serial serology looking for antibodies against pneumotropic bacteria and viruses. One of more infectious agents were shown in 47% of the episode studies of which 57% were exacerbations of COPD and treated 23% attacks of asthma. In the cases COPD bacteria were identified in 13 cases including Haemophilus influenzae [3], Streptococcus pneumoniae [3], Pseudomonas aeruginosa [3]. Amongst the 14 viruses recovered, the influenza virus [8] and the respiratory syncytial virus (VRS) [4] predominated. In 14 cases of acute asthma only 4 infectious agents were shown; Mycoplasma pneumoniae, influenza A, VRS and parainfluenza virus. The influenza virus was the agent most frequently discovered (26%) during the course of exacerbation of COPD and of asthma.

Bacteria

Evaluation of a blood culture medium BHI-S-Lysis for BCB Release System Roche.

The new blood culture medium BHI-S Lysis for the BCB Release System (F. Hoffmann-La Roche AG., Basel, Switzerland), which lyses blood cells, was compared with the Hémoline performance Duo (BioMérieux, Marcy l'Etoile, France) including a diphasic medium and a modified Wilkins Chalgren broth. A total of 1998 sets were each inoculated with 7 ml of blood. In the laboratory, agar-coated paddles were attached to the BHI-S-Lysis. Overall, there was a statistically significant difference between the BHI-S-Lysis and the diphasic medium, or Wilkins Chalgren broth, in the recovery of clinically important microorganisms. The yield of pathogens with the BHI-S-Lysis, diphasic medium and Wilkins Chalgren broth was 115, 68 and 58, respectively. Staphylococci were detected by the BHI-S-Lysis significantly more frequently. Out of 50 bacteraemias, 12 were diagnosed with the BHI-S-Lysis only, three with the diphasic medium only, and three with the Wilkins Chalgren broth only. Contaminant isolates occurred with significantly greater frequency in the BHI-S-Lysis than in the two BioMérieux systems, in particular with coagulase-negative staphylococci. No speed advantage was present for all groups of micro-organisms and blood culture media. This good performance of the BHI-S-Lysis medium in detecting a larger number of pathogens should be confirmed by further evaluations.

Bacteremia

Characterization of staphylococcal plasmids hybridizing with the fosfomycin resistance gene fosB.

The distribution of the fosB gene, coding for fosfomycin resistance, in 105 fosfomycin-resistant isolates of Staphylococcus from various geographical areas, was studied by Southern blot hybridization. Nucleotide sequences related to fosB were detected in 36 strains belonging to five species. Plasmids bearing fosB were often of a size similar to that of pIP1842 (2.54 kb) in S. epidermidis, most often small (2.4 to 4.1 kb) in other species including S. aureus where a 2.7-kb plasmid was found in 16 out of the 18 strains studied. The fosB gene was geographically dispersed since it was present in six different locations in France and also in Japan. The weak hybridization observed with plasmid DNA of certain strains of S. aureus, S. epidermidis, S. haemolyticus, S. saprophyticus, and S. warneri may indicate gene heterogeneity for fosfomycin resistance in Staphylococcus spp.

Blotting, Southern

Long-acting chloramphenicol versus intravenous ampicillin for treatment of bacterial meningitis.

In most developing countries, bacterial meningitis (BM) is associated with a high case-fatality rate. The search for a simple, convenient, and inexpensive antibiotic treatment remains a priority. In this study, a non-blinded, multicentre, randomised clinical trial of 528 cases of BM was done in two hospitals in Mali and Niger, between March, 1989, and May, 1990, to see whether a double injection of long-acting chloramphenicol (on admission to hospital and 48 h later) is as effective as a course of intravenous ampicillin (8 days, 4 times a day). The cumulative case-fatality rate on day 4 (principal end-point) among the chloramphenicol (254 patients) and ampicillin (274) groups were, respectively, 28% and 24.5% (relative risk 1.14, 95% confidence interval 0.86-1.52). No outbreak occurred during the study period. The hospital case-fatality rate was 33.1%. Main risk factors for death were associated with clinical condition on admission--ie, altered consciousness, convulsions, or dehydration. The case-fatality rates were 13% (21/161) for Neisseria meningitidis, 36.1% (48/133) for Haemophilus influenzae, and 67% (77/115) for Streptococcus pneumoniae. In a multiple logistic regression model, controlling for the differential distribution of potential risk factors (including bacterial species), there was no difference between treatment groups. Our findings suggest that long-acting chloramphenicol is a useful first-line presumptive treatment for BM in high-incidence countries.

Ampicillin