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Biomedical subjects

F Rousselet

Publications and source records attributed to F Rousselet.

At least 19 recordsLinked to original sources

Mitochondrial DNA polymorphisms: a study of 50 French Caucasian individuals and application to forensic casework.

The analysis of mitochondrial DNA polymorphisms has proved to be efficient on highly degraded samples or samples having little or no genomic DNA such as hair shafts. In order to use this very sensitive method, the authors first established a database by analysing the mitochondrial DNA polymorphism in 50 French white Caucasian individuals, applied the analysis to different types of samples that can be found in forensic investigations and finally performed this method on two forensic cases involving the discovery of highly putrefied unidentified remains.

Base Sequence↗

Cyclosporin-A inhibits human endothelial cells proliferation through interleukin-6-dependent mechanisms.

Cyclosporine A (CsA) is a widely used immunosuppressant which possesses significant side effects including nephrotoxicity and systemic arterial hypertension. In this work we evaluated the effects of CsA on human umbilical endothelial cell proliferation (HUVEC). Treatment of endothelial cells with CsA inhibited cell proliferation, and was correlated with an increase of interleukin-6 (IL-6) production and IL-6 mRNA expression. Addition of exogenous IL-6 also significantly altered cell proliferation. Furthermore, neutralization of endogenous IL-6 with a specific monoclonal antibody concomitantly with CsA treatment completely restored HUVEC proliferation. These results suggest that CsA-induced inhibition of HUVEC proliferation is mediated through an augmentation of IL-6 synthesis.

Antibodies↗

A pentaplex automated fluorescent typing system for forensic identification and French Caucasian population data.

The polymerase chain reaction (PCR) amplification of short tandem repeat (STR) loci has already proven to be a method of choice for large scale typing of DNA samples in which the conventional restriction fragment length polymorphism (RFLP) technique is ineffective. A quadruplex PCR including HUMvWFA31A, HUMF13A01, HUMTH01, and HUMFESFPS STR loci is used successfully for routine forensic applications in our laboratory. However, the need to increase the discrimination power of the PCR systems used prompted us to develop a second system of a pentaplex PCR for the analysis of 4 additional STR loci (HUMD8S1179, HUMD18S51, HUMD21S11, and HUMFIBRA) and the sex determination by amplification of a segment of the X-Y homologous Amelogenin gene. Allele and phenotype frequencies for these 4 STR systems were obtained by multiplex amplification, from approximately 200 randomly selected and unrelated French Caucasian individuals. Statistical calculations for these phenotype distributions met expectations for Hardy-Weinberg equilibrium. Furthermore, the French allelic frequencies of D18S51, D21S11, and HUMFIBRA loci were compared with the data obtained by the Forensic Science Service (UK) for the British Caucasian population and proved to be similar.

Alleles↗

Resistance of lipoprotein(a) to lipid peroxidation induced by oxygenated free radicals produced by gamma radiolysis: a comparison with low-density lipoprotein.

Lipid peroxidation of lipoprotein(a) [Lp(a)] by defined oxygen-centred free radicals (O2-/OH, O2-, O2-/HO2) produced by gamma radiolysis was compared with that of paired samples of low-density lipoprotein (LDL). Lp(a) appeared to be more resistant to oxidation than LDL, as indicated by the kinetic study of four markers of lipid peroxidation; decrease in vitamin E, formation of conjugated dienes and aldehydic products, and modification of electrophoretic mobility. In contrast, similar kinetics of lipid peroxidation were obtained for LDL and Lp(a-), which is the lipoparticle issued following the reductive cleavage of apolipoprotein(a) from Lp(a), thus suggesting that the greater resistance of Lp(a) to lipid peroxidation was due to the presence of apolipoprotein(a). Lipid peroxidation of Lp(a) and LDL induced by peroxyl radicals, which were produced by an azo compound [2,2'-azobis-(2-amidinopropane)dihydrochloride], confirmed both the resistance of Lp(a) to lipid peroxidation and the propensity of Lp(a-) to exhibit a greater susceptibility to oxidation than intact Lp(a). Our findings also indicated that the high content of apolipoprotein(a) in N-acetylneuraminic acid residues was only partly responsible for the resistance of Lp(a) to oxidation.

Amidines↗

French Caucasian population data obtained from fluorescently detected HUMvWFA31/A and HUMF13A01 short tandem repeat loci.

Allele and phenotype frequencies for two tetranucleotide STR (short tandem repeat or microsatellite) systems, HUMvWFA31/A and HUMF13A01, were obtained from a sample of approximately 240 unrelated individuals randomly selected from the French Caucasian population. PCR (polymerase chain reaction) products were analysed on 6% polyacrylamide denaturing gels and visualized using fluorescently labelled primers on the automated 373A ABI DNA sequencer (Applied Biosystems Inc.). French Caucasian allele frequencies were compared to other published Caucasian data. Conditions were optimised for the quadruplex PCR amplification of these two STR loci together with the HUMFESFPS and HUMTH01 loci and the quadruplex PCR was also performed on various forensic DNA samples.

Alleles↗

Enhanced susceptibility of low-density lipoprotein to in vitro oxidation in type 1 and type 2 diabetic patients.

Macrovascular disease represents a major cause of morbidity and mortality in patients with diabetes mellitus. Low-density lipoprotein (LDL) is involved in the pathogenesis of atherosclerotic lesions, through modifying processes such as oxidation. We examined the in vitro susceptibility to oxidation and the oxidizability of LDL isolated from the plasma of Type 1 and Type 2 diabetic patients. Two groups of diabetic patients (20 Type 1, 20 Type 2) were compared with sex- and age-matched non-diabetic control groups. In vitro oxidation of the purified LDL preparations was assessed by determination of the kinetics for the formation of conjugated dienes (lag phase duration, maximal rate and maximal dienes concentration) and by measurement of thiobarbituric acid-reacting substances (TBARS) in the presence of copper ions. LDL from both Type 1 and Type 2 diabetic patients exhibited a shorter lag phase duration for conjugated dienes formation (94 +/- 14 vs. 108 +/- 20 and 97 +/- 26 vs. 112 +/- 18 min for Type 1 and Type 2 diabetic groups vs. respective control groups, P < 0.05). We also observed an increase in maximal rate of conjugated dienes formation (2.21 +/- 0.55 vs. 1.52 +/- 0.31 and 2.02 +/- 0.55 vs. 1.52 +/- 0.31 nmol/mg LDL/min, P < 0.01) and of maximal production of TBARS (77.9 +/- 11.8 vs. 65.5 +/- 10.4 and 76.7 +/- 9.9 vs. 65.3 +/- 9.4 nmol/mg LDL protein, P < 0.05) in diabetic groups. Our results demonstrate both a higher susceptibility to oxidation and a higher oxidizability of LDL from diabetic patients, as much for Type 1 as Type 2 diabetic subjects with or without pre-existent vascular complications. This enhanced propensity of LDL oxidation in patients with diabetes mellitus could at least partly be attributable to quantitative and qualitative alterations in the chemical composition of LDL and to the glycoxidation process occurring on these lipoproteins.

Adult↗

Use of a renal tubule cell line (LLC-PK1) to study the nephrotoxic potential of a kappa-type Bence-Jones protein.

The cytotoxicity of a Bence-Jones protein was assessed using a porcine renal tubule cell line (LLC-PK1), with the aim of developing a model for studying the potential nephrotoxicity of these proteins. The effects of a kappa Bence-Jones protein on cell viability were studied by means of biochemical methods (supravital dye uptake and measurement of cellular enzyme activities) and morphological electron microscopy. After a 24-h-treatment with Bence-Jones protein, a moderate cytotoxicity (about 15%) was noted but only a minor difference compared to treatment with bovine albumin in the same conditions. The morphological study showed a few cells in the process of lysis, but their numbers were insufficient for the demonstration of a clear cytotoxic effect. Immunocytochemical studies showed Bence-Jones protein fixation on some cells, especially on the outer membrane. Labeling of the hyaloplasm and basal pole of a few cells pointed to internalization of protein by LLC-PK1 cells. Although the cytotoxicity of the Bence-Jones protein tested here was only moderate, the use of this model enabled its cytotoxic effect to be distinguished from that of beta-lactoglobulin. This isolate could serve as a "moderate control" for a later study with a BJP having caused acute renal failure.

Animals↗

Intestinal transport and processing of immunoglobulin G in the neonatal and adult rat.

The proximal intestine of neonatal rats expresses a specific receptor (RFcn) that binds immunoglobulin G (IgG) and is no longer expressed after weaning. The aim of this study was to quantify and compare the intestinal transport and processing of IgG in intestinal fragments with or without RFcn, with the fluid-phase transport of horseradish peroxidase (HRP). The mucosal to serosal transport and degradation of IgG and HRP were measured in neonatal and adult rats in vitro in Ussing chambers. IgG transcytosis occurred without degradation in the proximal intestine of neonatal rats, where RFcn is expressed, up to a luminal concentration of 300 micrograms/ml. At higher mucosal IgG concentrations, a degradative pathway was also involved. The immunoreactive IgG fluxes across the proximal intestine of neonatal rats were higher than those observed in the distal neonatal intestine or those in the proximal and distal adult intestine. The rate of HRP transcytosis was higher than that of IgG but it involved a mainly degradative pathway. These results suggest that in the proximal intestine of the neonatal rat, where RFcn is expressed, the transcytotic rate for IgG is not increased, but the nondegradative transport of immunoreactive IgG is favored, especially at low luminal concentrations.

Aging↗

Discrepancies between lipoprotein(a) concentrations in icteric sera measured by immunonephelometry and electroimmunodiffusion.

We compared the lipoprotein(a) [Lp(a)] levels in 32 icteric sera determined both by an electroimmunodiffusion assay (EIA), using the Hydragel Lp(a) kit (Sebia, France) and by two immunonephelometric assays, one on a Behring Nephelometer Analyzer (BNA), using antiserum from immunofrance, and the other on a Beckman analyzer (Array), using antiserum from Dako (Denmark). With the EIA assay, the Lp(a) level was 0.09 +/- 0.09 (mean +/- SD in g/L), with the BNA assay, 1.01 +/- 1.51 and with the Array assay, 0.05 +/- 0.05. Sample blanks values (0.76 +/- 1.28 g/L) demonstrated that the high Lp(a) levels obtained in the BNA assay are caused by nonspecific precipitation. Analysis of the precipitate indicated the presence of Lipoprotein X, an abnormal lipoprotein that appears in the serum of patients with obstructive jaundice or with lecithin-cholesterol acyltransferase deficiency. The precipitant seems to be polyethyleneglycol (PEG) that was added to the reaction medium in both the BNA and the Array assays to stabilize the Lp(a)-anti Lp(a) immune complex. In the Array assay, interference by this nonspecific precipitation is eliminated by preliminary centrifugation of the diluted sample. However, coprecipitation of Lp(a) could occur during this step. Consequently, the results of Lp(a) measurement in serum from patients with hepatobiliary diseases should be interpreted with caution when immunonephelometric assays are used with a medium containing PEG.

Humans↗

Electrophoretic study of the physico-chemical characteristics of Bence-Jones proteinuria and its association with kidney damage.

AIM: To identify a physico-chemical criterion, or set of criteria, explaining and possibly predicting the nephrotoxic behaviour of Bence-Jones proteins (BJP). METHODS: The electrophoretic mobility and isoelectric point (pI) of 92 BJP isolates were determined using various electrophoresis procedures on polyacrylamide gel. The proportions of monomers and dimers were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE) in 58 cases. PAGE data for 10 BJP isolates were used to construct Ferguson plots and titration curves. RESULTS: The distribution of electrophoretic mobility and pI values was bimodal and showed a positive correlation when the pI was above 6. The values of these two parameters in 22 patients with renal impairment were not significantly different from those in the patients without renal impairment, and the statistical analysis showed no predictive value for the onset of renal impairment. However, patients excreting the lambda light chain isotype had a 2.8-fold higher risk of developing renal impairment compared with the other patients. Studies of the charge variation of the protein with pH indicated three types of behaviour, suggesting that the charge of BJP is highly variable at physiological pH. CONCLUSION: It is important to study not only the positivity or negativity of the BJP charge at a given pH, but also its intensity. The study of the BJP titration curves in patients with renal impairment suggests that a low charge at physiological urinary pH could predict renal impairment.

Aged↗

A sequential and simple determination of zinc, copper and aluminium in blood samples by inductively coupled plasma atomic emission spectrometry.

A sequential method of measuring zinc, copper and aluminium in serum by inductively coupled plasma (ICP) spectrometry is described. It involves a 1/5 dilution of serum with a potassium chloride solution which enhances aluminium signal intensity and reduces variations between different matrix compositions. The method is as sensitive as atomic absorption for zinc (sensitivity: 0.11 mumol/l) and copper (sensitivity: 0.020 mumol/l) and can also be applied to monitor aluminium (sensitivity: 0.12 mumol/l) for patients receiving total nutrition therapy or hemodialysis. Its linearity extends at least to 200 mumol/l for copper and zinc and to 20 mumol/l for aluminium. The correlations with atomic absorption are satisfactory for the 3 parameters, as assessed by the correlation coefficients established for both methods. A reference interval was established with 34 sera of control subjects (19 men, 15 women) which showed an average zinc, copper and aluminium of 14.5 (S.D. 2.6), 17.3 (S.D. 2.1) and 0.32 (S.D. 0.12) mumol/l, respectively. This method does not require a simultaneous ICP spectrometer and can be performed with 1 ml of serum in a single tube, using a routine sequential ICP spectrometer.

Aluminum↗

Lipoprotein(a) in diabetic patients with and without chronic renal failure.

OBJECTIVE: To examine the distribution of Lp(a) plasma levels in patients with IDDM and NIDDM, and in nondiabetic and IDDM patients with chronic renal failure. RESEARCH DESIGN AND METHODS: Cross-sectional study of Lp(a) plasma levels in a population of diabetic patients with stable metabolic control, with simultaneous determination of plasma lipids, fasting plasma glucose, and HbA1. Thirty-six patients with IDDM, 90 with NIDDM, and 41 with chronic renal failure (20 IDDM, 21 nondiabetic) were compared with 78 control subjects. RESULTS: Lp(a) plasma levels were significantly higher in IDDM and NIDDM patients, as well as in nondiabetic and IDDM patients with chronic renal failure compared with control subjects. No correlation was observed between Lp(a) and lipid plasma levels, fasting plasma glucose, and HbA1. CONCLUSIONS: Lp(a) may contribute to the increased prevalence of atherosclerotic disease in diabetic patients and patients with chronic renal failure, especially in IDDM patients whose lipoprotein pattern was not different from that of the control group.

Adult↗

Comparison of HbA1 and fructosamine in diagnosis of glucose-tolerance abnormalities.

Total glycosylated hemoglobin (HbA1) and fructosamine were evaluated as screening tools for detection of glucose-tolerance abnormalities in 144 asymptomatic subjects undergoing a 75-g oral glucose tolerance test. Subjects were classified according to World Health Organization criteria as having normal (n = 78), impaired (n = 40), or diabetic (n = 26) glucose tolerance. We found good specificity for HbA1 and fructosamine (100 and 97%, respectively) but low sensitivity (15 and 19%, respectively). At the intersection of the curves of sensitivity and specificity drawn from various thresholds of normality, both sensitivity and specificity were 75% for HbA1 and 55% for fructosamine. Thus, neither HbA1 nor fructosamine seems to be suitable for the diagnosis of mild abnormalities in glucose tolerance.

Blood Glucose↗

Comparison of fructosamine with glycated hemoglobin as an index of glycemic control in diabetic patients.

Fasting and postprandial (or post-glucose load) plasma glucose, total HbA1 and fructosamine (F) were simultaneously assessed in 371 diabetic patients (125 insulin dependent and 246 non-insulin dependent) and in 122 nondiabetic subjects, (98 with normal glucose tolerance and 24 with impaired glucose tolerance). Fructosamine yielded nearly similar information as HbA1 about glycemic control, since similar relationships were observed between plasma glucose values and HbA1 or fructosamine levels in the different groups. A longitudinal study performed during a three-month follow-up in 74 diabetic patients and extended to six months in 19 of them, without any modification of treatment, indicated that reproducibility of HbA1 and fructosamine was nearly the same with a slight advantage for HbA1. The only clinically significant difference results from the longer half-life of hemoglobin when compared to serum proteins. Fructosamine assay should be proposed as a complement of HbA1 in the management of diabetic patients when detection of recent metabolic changes is needed.

Adult↗

Relationship between hair, serum and bone aluminium in hemodialyzed patients.

To establish the predictive value in osteodystrophy of hair aluminium content in patients on hemodialysis, we compared the aluminium levels in serum, cortical and trabecular bone and hair of 40 such patients with the levels in 23 healthy controls. In the patients, mean hair aluminium content was significantly higher than the controls (0.226 mumol/g, range: 0.09-0.500 mumol/g, n = 39 versus 0.126 mumol/g, range: 0.020-0.330 mumol/g, n = 49) but there was a large overlap between the two groups. The patients exhibited a significant correlation between serum and cortical bone aluminium measured by atomic absorption spectrometry (rs = 0.67, n = 37, p less than 0.05), but no correlation between aluminium in hair and in serum or bone, whether cortical or trabecular. Bone histomorphometric studies also indicated that unlike aluminium levels in cortical or trabecular bone, its levels in hair are not predictive of aluminium-induced osteomalacia. Consequently, hair aluminium cannot suitably replace bone and serum aluminium as a criterion of osteodystrophy in hemodialyzed patients.

Adult↗