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Biomedical subjects

F Lefèvre

Publications and source records attributed to F Lefèvre.

At least 37 records · Page 2Linked to original sources

[Upper urinary tract MRI].

New MRI modalities today available allow exploration of many kidney diseases. Main developments have been made in the diagnosis of small tumors and tumoral extension of huge masses. The other important way of application is MR urography which perfectly show the dilated urinary tract in few seconds. Their potential application are for renal insufficiency, allergic reaction to iodine contrast media and in cases of contra indication to ionised radiations. It is a complementary sequence to assess the urinary tract in pelvic exploration. The evaluation of kidney function will be possible in the near future.

Contraindications↗

Enhanced protective response and immuno-adjuvant effects of porcine GM-CSF on DNA vaccination of pigs against Aujeszky's disease virus.

This study was conducted to investigate whether the co-delivery of DNA encoding porcine cytokines would enhance a protective immune response in pigs to a Pseudorabies virus (PRV; or Aujeszky's disease virus) DNA vaccine. Aujeszky's disease in pigs results in respiratory and nervous symptoms with important economic losses. To evaluate cytokine effects, eukaryotic expression vectors were constructed for porcine GM-CSF, IL-2 and IFN-gamma. cDNA for each of these cytokines was inserted under the control of a CMV promoter in the pcDNA3 plasmid and cytokine expression was confirmed after DNA transfection in various mammalian cell cultures by bioassays (GM-CSF and IL2) and ELISA (IFN-gamma). Pigs were vaccinated by single intramuscular injection with plasmid DNA encoding PRV gB and gD along with various combinations of cytokine plasmid constructs. Pig serum was tested for the production of antibody by isotype specific anti-PRV ELISA. Pigs were then challenged with the highly virulent PRV strain NIA3 on day 21 after vaccination. The survival and growth rate of pigs were monitored for seven days after the viral challenge. The co-administration of GM-CSF plasmid increased the immune response induced by gB and gD PRV DNA vaccine. This immune response was characterized by an earlier appearance of anti-PRV IgG2, a significantly enhanced anti-PRV IgG1 and IgG2 antibody response, a significantly decreased and shortened viral excretion in nasal swabs and an improved protection to the viral challenge. In contrast, the co-administration of porcine IL-2 or IFN-gamma had no adjuvant effects. Our results thus demonstrate for the first time that the application of porcine GM-CSF gene in a DNA vaccine formulation can exert immuno-adjuvant and protective effects with single vaccination in the natural host pig against Aujeszky's disease.

Adjuvants, Immunologic↗

[Computed tomography study of the sigmoid colon: discriminating diagnostic criteria and interobserver correlations].

PURPOSE: To assess the value of pericolonic findings at CT in the evaluation of the sigmoid colon. MATERIALS AND METHODS: A total of 210 CT examinations were retrospectively reviewed by 3 blinded radiologists. Data was analyzed to determine the interobserver correlation and the value of pericolonic and colonic wall findings in diagnosis of sigmoid colon pathology. RESULTS: The interobserver correlation for pericolonic findings was equal to or superior to that for colonic wall findings. The presence of abnormal pericolonic fat was the most sensitive (88%) and specific (93%) sign to differentiate a diseased sigmoid colon from a normal one or from sigmoid diverticulosis. Wall-thickening was less sensitive (82%) and specific (76%). Findings suggesting malignancy over diverticulitis included acute zone of transition, focal fatty infiltration, and lymph nodes. Symmetrical and circumferential wall thickening, target-like enhancement, and local fatty proliferation were findings suggesting colitis over diverticulitis. Wall thickening more than 15 mm, involvement of 15 cm or less, asymmetrical involvement, acute zone of transition, and homogeneous or heterogeneous enhancement were findings suggesting malignancy over colitis. CONCLUSION: To render a diagnosis, the evaluation of the fat infiltration must prevail on the parietal thickening appreciation.

Adipose Tissue↗

[Marijuana, health, disease, and freedom: analysis of an Internet forum].

This paper takes a Health Education perspective to analyze a debate forum on the Brazilian Internet site entitled "Universo On-Line", in which the following questions were addressed: "Do you believe that marijuana is harmful to one's health?" "In your opinion, should marijuana use be decriminalized?" By applying qualitative discourse analysis techniques to responses from the forum, we were able to identify six main types of discourse, reflecting the opinions of six "collective subjects" concerning drugs, health, disease, and freedom and existing as social representations in the current Brazilian collective imagination. Research on these social representations allows one to establish criteria for intervention in the field of Health Education.

Alcoholism↗

Decreased number of nucleolar organizing regions in collagen lattice cultured fibroblasts.

Dermal fibroblasts cultivated in tridimensional matrices (lattices) of collagen exhibit a very low metabolic activity, and a low protein synthesis in particular. We have previously shown that ribosomal RNA content and half-life were decreased in collagen lattice cultured fibroblasts when compared to monolayer cultured fibroblasts. In this study, we seeded fibroblasts in collagen lattices and investigated the influence of matrix on the number of nucleolar organizing regions. We found that fibroblasts in fully retracted lattices exhibited a significant decrease of 45% (P < 0.001) in the number of nucleolar organizing regions when compared to monolayer cultured fibroblasts. This decrease was correlated to the decrease in ribosomal RNA content. These data suggest that extracellular matrix induces early alterations of synthesis and/or processing of ribosomal RNAs, explaining, at least partly, the resulting low metabolic activity.

Adult↗

Intrauterine infusion of high doses of pig trophoblast interferons has no antiluteolytic effect in cyclic gilts.

In the pig species, the preimplanting trophoblast is known to synthesize and secrete high amounts of interferon during early development. Previous experiments in cyclic gilts using total conceptus secretory proteins suggested that porcine trophoblastic interferons, unlike those of ruminants, exert no effect on the luteal cycle. In the present experiment, cyclic Meishan gilts were divided into two groups, cannulated on both uterine horns, and given daily injections of either a placebo or increasing doses of a mixture of recombinant interferon-gamma and interferon-delta, on Days 11-14 of the estrous cycle. In treated gilts, the injected doses were much higher than those previously found in uterine perfusates from pregnant gilts. However, no significant differences could be found between the control (n = 4) and the treated (n = 5) group concerning the days of the estrous cycle for mid-decrease of progesterone (control: Day 14.5+/-0.57 [mean+/-SD]; treated: Day 15+/-1.25), the day of estrus (control: Day 19+/-0.96; treated: Day 19.6+/-0.55), and the subsequent ovulation rate (control: 14+/-2.2 corpora lutea; treated: 13.1+/-1.1 corpora lutea). These data confirm that pig trophoblastic interferons, unlike those of ruminants, do not themselves exert an antiluteolytic effect. A possible synergistic effect of embryonic estrogens on the luteal functions of nonpregnant sows remains to be determined.

Animals↗

Long-term variability of serum lipoprotein(a) concentrations in healthy fertile women.

Lipoprotein(a) is a unique lipoprotein with atherothrombogenic properties. Although its blood concentration is mainly genetically determined, various factors exist which may cause variability. These may influence the clinical use of the results. We studied lipoprotein(a) biological variation by a rate nephelometric assay over a period of two years in a population of healthy fertile women. The study was performed in 12 volunteers, healthy subjects with various lipoprotein(a) concentrations, by monthly determinations during one year and a single determination one year later, together with measurements of total, high density lipoprotein and high density lipoprotein2 cholesterol, triglycerides and apolipoproteins A1 and B. The intra-individual variability of lipoprotein(a) ranged between 4 to 20%, with three subjects showing a coefficient of biological variation higher than 15%. In absolute terms, the difference between two determinations could represent 0.44 g/l or 50% of the mean value. This study suggests that physiological lipoprotein(a) variations should be taken into account for clinical purposes, especially in patients in need of thorough risk evaluation.

Adult↗

[Single shot fast spin echo sequence MRI cholangiopancreatography].

PURPOSE: To assess the value of single shot fast spin echo MR sequence (SS-FSE) in the morphological analysis of the biliary tree and pancreatic ducts and to compare its accuracy with other imaging methods. MATERIAL AND METHODS: 95 consecutive patients referred for clinical and/or biological suspicion of biliary obstruction were explored with MR cholangiopancreatography (MRCP). All patients were explored with a Signa 1.5 T GE MR unit, with High Gradient Field Strength and Torso Phased Array Coil. Biliary ducts were explored with SS-FSE sequence, coronal and oblique coronal 20 mm thick slices on a 256 x 256 matrix. Total acquisition time was 1 second. Native pictures were reviewed by two radiologists blinded to clinical information. In case of disagreement, a third radiologist's judgement was requested. In 88 cases, MRCP results were compared with direct biligraphy methods. RESULTS: In all cases, MRCP produced high quality images without MIP or other post-processing methods. For detection of biliary tree distensions, the concordance value of MRCP was over 91% (Kappa 0.82). For detection of biliary tree and/or pancreatic duct obstruction, MR sensitivity was 100% and specificity 91%. The overall diagnostic concordance value of MRCP was > or = 93%. Difficulties in MRCP were caused by functional diseases or benign stenosis. MRCP accurately diagnosed all lithiasic obstructions starting from a stone size of 3 mm. CONCLUSION: MRCP produces fastly high-quality images. As it is totally safe, it can be proposed as a first intention method in biliopancreatic duct explorations.

Adolescent↗

Alanine-stretch scanning mutagenesis: a simple and efficient method to probe protein structure and function.

We have developed a foolproof method to substitute a stretch of residues by alanines. After the introduction of aPstI site by IPCR, thus creating two alanine codons, additional codons are introduced at this site through the use of an 'alanine-stretch cartridge'. These cartridges comprise an antibiotic resistance gene flanked on both sides by alanine codons. Excision of the resistance gene byPvuII then yields the correct insertion of codons. The method is both highly reliable and flexible and should be of general use.

Alanine↗

Topographical and functional investigation of Escherichia coli penicillin-binding protein 1b by alanine stretch scanning mutagenesis.

Penicillin-binding proteins (PBPs) are the targets of beta-lactam antibiotics. We have used a systematic five-alanine substitution method (called ASS [alanine stretch scanning] mutagenesis) to investigate the functional or structural role of various stretches of amino acids in the PBP1b of Escherichia coli. To probe the specific activity of each variant, the antibiotic discs assay was used with strain QCB1 (delta ponB) in the presence of cefaloridine, which totally inhibits the complementing action of PBP1a. This in vivo test has been combined with a quick and efficient in vitro test of the penicillin-binding activity of each of these variants with fluorescent penicillin. This approach has enabled us to show an unexpected role of the N-terminal and C-terminal tails of PBP1b. Moreover, we have established the correct position in PBP1b of the SMN motif that, with the SXXK and the KTG motifs, constitutes the signature of the penicilloyl serine transferases family. Finally, we have shown that the transglycosylase and the transpeptidase domains are separated by an inert linker region, where substitutions and insertions can be made without hindering the in vivo and in vitro activity of the protein.

Alanine↗

Modulation of protein synthesis by extracellular matrix: potential involvement of two nucleolar proteins, nucleolin and fibrillarin.

Fibroblasts cultivated in a collagen matrix exhibit a large decrease in the synthesis of most proteins, depending on transcriptional and posttranscriptional controls. We have previously shown that ribosomal RNA content and half-life were decreased in collagen lattice cultures. Here, we cultivated human dermal fibroblasts in monolayers and in lattices and studied by competitive RT-PCR analysis the expression of the nucleolar proteins nucleolin and fibrillarin, two key factors in ribosome processing and association. Nucleolin expression was found increased, and fibrillarin expression decreased, in collagen-lattice vs monolayer-cultured fibroblasts, with some variability according to the strains (+25 to +250% and -40 to -60%, respectively). These data suggest that a possible trouble of the association between neosynthesized rRNA and nucleolar proteins is, at least partly, responsible for the inhibition of protein synthesis induced by the extracellular matrix.

Cells, Cultured↗

Molecular genetics of rust resistance in poplars (Melampsora larici-populina Kleb/Populus sp.) by bulked segregant analysis in a 2 x 2 factorial mating design.

With random amplified polymorphic DNA (RAPD) markers, we have tagged a genomic region in Populus sp. involved in qualitative resistance to Melampsora larici-populina. Our approach was based on three steps: use of RAPD markers that can be quickly and efficiently researched: application of "bulked segregant analysis" technique on individuals of one interspecific family P. trichocarpa x P. deltoides to search for RAPD markers linked to resistance; and validation of these markers in two other families linked with the first one in a 2 x 2 factorial mating design. Of five detected markers, only one marker M03/04_480 was polymorphic in the three segregating families, involving 89 individuals and four different parents. We have estimated the recombination value of 1 cM with 1 cM sampling error.

Basidiomycota↗

The asparagine to aspartic acid substitution at position 276 of TEM-35 and TEM-36 is involved in the beta-lactamase resistance to clavulanic acid.

TEM-35 (inhibitor resistant TEM (IRT)-4) and TEM-36 (IRT-7) clavulanic acid-resistant beta-lactamases have evolved from TEM-1 beta-lactamase by two substitutions: a methionine to a leucine or a valine at position 69 and an asparagine to an aspartic acid at position 276. The substitutions at position 69 have previously been shown to be responsible for the resistance to clavulanic acid, and they are the only mutations encountered in TEM-33 (IRT-5) and TEM-34 (IRT-6). However, the N276D substitution has never been found alone in inhibitor-resistant beta-lactamases, and its role in resistance to clavulanic acid was thus unclear. The N276D mutant was constructed, purified, and kinetically characterized. It was shown that the substitution has a direct effect on substrate affinities and leads to slightly decreased catalytic efficiencies and that clavulanic acid becomes a poor substrate of the enzyme. Electrospray mass spectrometry demonstrated the simultaneous presence of free and inhibited enzymes after incubation with clavulanic acid and showed that a cleaved moiety of clavulanic acid leads to the formation of the major inactive complex. The kinetic properties of the N276D mutant could be linked to a salt-bridge interaction of aspartic acid 276 with arginine 244 that alters the electrostatic properties in the substrate binding area.

Base Sequence↗