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Biomedical subjects

B Hainque

Publications and source records attributed to B Hainque.

66 records · Page 4Linked to original sources
PubMed1988

Rabbit articular chondrocytes: an in vitro model for studying the effect of sodium aurothiopropanol sulfonate on proliferation kinetics, type II collagen phenotype and mitochondrial activity.

Despite the benefits of chrysotherapy the responsible mechanism of action of gold compounds remains unclear. At a concentration of 5 x 10(-4) M, sodium aurothiopropanol sulfonate (SAS) modified the in vitro proliferation kinetics of articular chondrocytes by reducing growth, viability and plating efficiency. Flow cytometry analysis, using propidium iodide DNA staining, revealed slight but significant cell arrest in G2+M which, in fact, represents an increase in the proportion of binucleate cells. SAS did not induce any variations in chondrocyte phenotype stability as far as the biosynthesis of type II collagen was concerned, and no appreciable changes in overall mitochondrial activity reflected by rhodamine 123 incorporation.

Animals↗
PubMed1987

Glucocorticoid binding during the differentiation of 3T3-F442A fibroblasts into adipocytes. A possible regulatory effect of insulin.

Until recently, few studies had been carried out on receptors for glucocorticoids in adipocytes, although the role of these steroids is considerable. In the present studies, we chose the pre-adipocyte line 3T3-F442A, which constitutes an excellent model for investigating the differentiation and function of adipocytes. Using a whole cell assay system, we showed the existence of a homogenous class of sites with the characteristics of glucocorticoid receptors, that is, high-affinity binding which is reversible, specific and saturable. Whatever the state of cellular differentiation, the affinity of the receptor for dexamethasone did not vary, although we observed an increase in the number of sites during differentiation. When cells were differentiated in the presence of insulin, there was a further increase in the binding capacity; moreover, insulin deprivation of such adipocytes caused a decrease in the number of sites. Our results therefore suggest that factors other than the glucocorticoids themselves influence dexamethasone binding. It is suggested that insulin plays a role in the regulation of the number of glucocorticoid receptors.

Adipose Tissue↗
PubMed1987

Effects of dexamethasone on the growth of cultured rabbit articular chondrocytes:relation with the nuclear glucocorticoid-receptor complex.

This study reports that dexamethasone at a high dose (10(-4) mol/l) induced slowing of the in vitro proliferation of rabbit articular chondrocytes in both monolayer and clonal culture. This effect is consistent with an inhibition of DNA and RNA synthesis and was characterised by an accumulation of cells in the G0G1 phase of the cell cycle, as shown by flow cytometric analysis. Therefore we determined the extent of nuclear localisation of dexamethasone-receptor complexes. The results showed a discrepancy between 50% growth inhibitory dose (10(-4) mol/l) and the apparent affinity, KD (1.4 (SD 0.2) X 10(-9) mol/l). Thus the growth inhibition of rabbit articular chondrocytes by dexamethasone did not seem to be related exclusively to an interaction with the glucocorticoid-receptor complexes.

Animals↗
PubMed1986

G2 arrest, binucleation, and single-parameter DNA flow cytometric analysis.

One important facet of flow cytometry involves the effects of pharmacological agents on cell cycle progression. Comparative G2 fraction perturbations were examined: effects of sodium butyrate on articular chondrocytes, effects of an antineoplastic agent (SOAZ) and an antirheumatic drug (D-penicillamine) on HeLa cells. Even though DNA flow cytometric analysis detects preferentially an induction of G2 arrest, the mode of action of these agents on the cell cycle is different. Sodium butyrate and D-penicillamine lead to an increase of binucleate cells due to cytokinesis perturbation. Because of similar fluorescence intensity, distinguishing G2 from binucleate GO/1 cells is not easily possible using DNA content measurement and reflects a failure of flow cytometry in the detection of binucleate cells. Rapid cell cycle analysis of single cells should contribute greatly to the study of pharmacological interactions, but DNA flow cytometric measurements obtained from cultured cells exposed to certain agents must be cautiously interpreted because those may interact on cytokinesis and induce artefacts in histogram interpretation.

Animals↗
PubMed1985

[Course of the concentration of serum free amino acids as a function of time and the method of preservation].

In order to determine the optimal conditions for storage in the case of deferred serum amino acid assay, the authors studied the effect of storing the sample for four hours at two different temperatures: melting ice and ambient temperature, as well as the effect of storing the amino acids for three months according to four methods of storage; freezing at -20 degrees C and -80 degrees C of deproteinised and non-deproteinised serum. When the analysis was performed within four hours of collection of the specimen, the best conditions are obtained when the sample is kept at the temperature of melting ice. The optimal conservation consists of immediately deproteinising the sample and storing it frozen for 14 days.

Adult↗
PubMedDate not supplied

Peroxisome proliferator activated receptor-gamma, leptin and tumor necrosis factor-alpha mRNA expression during very low calorie diet in subcutaneous adipose tissue in obese women.

BACKGROUND: PPAR gamma, leptin and TNF alpha are three major factors that play a key role in influencing adipocyte differentiation and both adipose tissue function and metabolism. However, the regulation of these three genes during a dynamic period of weight loss is unknown. We therefore investigated the concomitant regulation of the mRNA expression of PPAR gamma, leptin and TNF alpha in adipose tissue during a 21-day very low calorie diet (VLCD) in 12 non-diabetic obese women. METHODS: The mRNA levels of PPAR gamma, leptin and TNF alpha were quantified by quantitative RT-competitive PCR in abdominal subcutaneous adipose tissue before and during VLCD (940 kcal/day). RESULTS: VLCD induced weight loss (approximately 6 kg) and improved insulin sensitivity. Simultaneously, VLCD induced the reduction in the adipose tissue mRNA abundances of PPAR gamma (-13%, p < 0.05) and of leptin (-58%, p < 0.005), whereas TNF alpha mRNA levels increased (+78%, p < 0.005). PPAR gamma and leptin mRNA levels were correlated before (r = 0.778, p < 0.01) and after VLCD (r = 0.797, p < 0.01). Serum HDL-cholesterol concentrations were positively associated with PPAR gamma (r = 0.696, p < 0.03) and leptin (r = 0.806, p < 0.01) mRNA levels. CONCLUSIONS: The increase in TNF alpha mRNA levels suggested that a local increased expression of this cytokine in adipose tissue might play a role in the control of the fat mass during weight loss. PPAR gamma and leptin mRNA levels were positively associated both before and after VLCD, suggesting that common regulatory mechanism(s) might control their expression. More strikingly, we found strong positive correlations between circulating HDL-cholesterol and both PPAR gamma and leptin mRNA levels, suggesting the existence of physiological links between circulating lipoprotein metabolism and adipose tissue function.

Abdomen↗
PubMedDate not supplied

Relationship between plasma plasminogen activator inhibitor 1 and insulin resistance.

High plasminogen activator inhibitor 1 (PAI-1) levels are associated with an increased cardiovascular risk of atherothrombosis. Furthermore, increased plasma PAI-1 levels are associated with dyslipidemia, hyperinsulinemia and hypertension. This association between PAI-1 and metabolic components of the Metabolic Syndrome could explain the predisposition of insulin resistant patients to atherothrombosis. Recent studies have suggested that visceral adipose tissue might be the link between elevated plasma PAI-1 and insulin resistance in the Metabolic Syndrome. Indeed, visceral adipose tissue was proposed as a potentially important source of PAI-1 in humans. However, in light of recent studies, visceral adipose tissue appears to be involved in the increase of plasma PAI-1 via the metabolic disorders usually associated with central obesity, rather than directly. High plasma PAI-1 levels are undoubtedly related to insulin resistance, and the mechanisms which could explain such an increase in the Metabolic Syndrome appear to be multi-factorial and remain to be elucidated. These mechanisms may involve several metabolic disorders such as hyperinsulinemia, dyslipidemia, impaired glucose tolerance and hypertension, which would favor PAI-1 synthesis and release from different cell types.

Adipose Tissue↗
PubMedDate not supplied

Percutaneous adipose tissue biopsy by mini-liposuction for metabolic studies.

BACKGROUND: Large fat-biopsy samples are necessary for extensive simultaneous assays. Subcutaneous adipose tissue is generally obtained either by needle-biopsy aspiration (approximately 1 g) or by open surgical biopsy for larger samples. We propose a reliable technique that permits the removal of large adipose tissue samples percutaneously. METHODS: A vertical 5-mm slit was made in the abdominal skin of 15 obese patients, and a liposuction mini-cannula (10 cm x 3 mm) attached to a 20-mL sterile plastic syringe was inserted. A vacuum was created by withdrawing the syringe plunger and fixing it with a specially adapted clamp. RESULTS: We obtained 3- to 15-g samples of adipose tissue without conspicuous scarring or visible contour changes. A significant correlation between the body mass index and the amount of adipose tissue was observed (r = .50, p < .05). Isolated adipocytes remained highly viable, as evidenced by glucose transport under both basal and maximal insulin-stimulated conditions. The mini-liposuction procedure was fast, and once the mini-cannula was removed from the wound, patients could immediately move about freely. CONCLUSION: The new technique described here for fat biopsy using a mini-cannula is less traumatic than open surgical biopsy, and it permits the removal of larger adipose tissue samples than do the usual biopsy techniques. Furthermore, adipose cells are removed intact, and the simplicity of the method facilitates repeat biopsies.

Adipose Tissue↗
PubMedDate not supplied

[Influence of pyridoxal phosphate in measuring aminotransferases activities in patients with viral hepatitis].

Effect of a pyridoxal phosphate (PP) supplementation of reagents used for ALT and AST measurement was studied in serum of 20 patients suffering from viral hepatitis. Measurements of enzyme activities were carried out at 37 degrees C, using an automate (AU 600, Olympus). Significant differences (p < 0.0001) were observed both for ALT and AST, meanwhile they were more marked for ALT than for AST. This difference was associated with a strong interindividual variability regarding PP activation effect on ALT. In conclusion, aminotransferase measurements should be carried out with a reagent supplemented with PP, when the enzyme activity is used to evaluate a cytolysis. The same recommendation applies when ALT results are integrated into various combinations developed for the evaluation of liver status.

Adult↗
PubMedDate not supplied

[Results transferability on RXL, ARX, X-Pand, BN2 (Dade Behring) and modular DP (Roche Diagnostics) analysers: application to component assays of fibrotest and Actitest].

The follow up of patients with chronic liver diseases and the data from multicentric clinical studies are affected by the variability of assay results for the same parameter between the different laboratories. Today, the main objective in clinical chemistry throughout the world is to harmonise the assay results between the laboratories after the confirmation of their traceability, in relation to defined reference systems. In this context, the purpose of our study was to verify the homogeneity of haptoglobin, apolipoprotein A1, total bilirubin, GGT activity, ALAT activity results, which are combined in Fibrotest and Actitest, between Dimension Analysers RXL, ARX and X-PAND (Dade Behring Society). Moreover, we verified the transferability of Fibrotest and Actitest results between the RXL, and either the BN2 (haptoglobin and apolipoprotein A1) or the Modular DP (total bilirubin, GGT and ALAT activity concentrations). The serum samples from 150 hospitalised patients were analysed on the different analysers. Specific protein assays were calibrated using solutions standardised against reference material on Dimension and BN2 analysers. Total bilirubin assays were performed by a diazoreaction on Dimension and Modular DP analysers. The GGT and ALAT activity measurements on the Dimension analysers were performed in accordance with the reference methods defined by the International Federation of Clinical Chemisty and Laboratory Medicine (IFCC). On the Modular, enzyme activity measurements were performed according to the Szasz method (L-gamma- glutamyl-4-nitroanilide as substrate) modified by Persijn and van der Slik (L-gamma- glutamyl-3-carboxy- 4-nitroanilide as substrat) for GGT and according to the IFCC specifications for ALAT. The methods of enzymatic activity measurement were calibrated on the Modular only. Liver fibrosis and necroinflammatory activity indices were determined using calculation algorithms, after having adjusted each component's result of Fibrotest and Actitest for gender and age. Our study has shown, for each parameter, harmonious results between the Dimension analysers and between RXL and BN2- Modular DP. Disregarding alpha-2 macroglobulin which cannot be assayed on RXL, the results of Fibrotest and Actitest were similar as performed on BN2- Modular DP and RXL.

Blood Chemical Analysis↗
PubMedDate not supplied

[Alpha 2 macroglobulin immunoturbidimetric assays (DakoCytomation reagents) on Roche Diagnostic analysers (Modular P, Cobas Integra). Application to FibroTest-Actic-Test].

BACKGROUND: Alpha2Macroglobulin (A2M) measure showed a revival since it was introduced into FibroTest-ActiTest-Fibro (FT-AT-Fibro) algorithm. More often than not, this assay is performed in immunonephelemetry. Progresses in the comprehension of fibrosis dynamics and better treatment efficacy follow-up will increase FT-AT-Fibro prescriptions. Despite efforts to standardize methods of enzymatic activity measure and proteins measure, we still observe important interlaboratory and intersystem variability. AIM: The primary aim of the study is to validate immunoturbidimetric measure of A2M on Modular P and Cobas Integra analysers (Roche Diagnostics) in utility channel using DakoCytomation reagents in order to extend the analytical system range allowed to measure A2M. The secondary aim of the study is to verify transferability of the six FT-AT composants (A2M, haptoglobin, apolipoprotein A1, total bilirubin, GGT and ALT) to Roche Diagnostics equipment by comparing with results measured on the reference system. RESULTS: A2M measures (n = 146) showed linearity, repetitiveness and were reproducible. Readjustments to adapt A2M measures were required. A corrector factor of 0.84 for Modular P and of 0.87 for Cobas Integra was introduced in order to readjust the immunoturbidimetric method to the immunonephelemetric method. The rationale of proposed corrector factors is based on the use of Dade Behring and DakoCytomation reagents (antisera and calibrant). Biologist vigilance is required to point out modifications or variations in reagents that could be done by the company. The six parameters results transferability from the reference system to Roche Diagnostics was demonstrated by statistic analysis. FT-AT showed excellent correlations to the reference system for Modular P and Cobas Integra analysers. In this study no difference more than 0.11 was recorded and only few subjects had differences between 0.05 and 0.10. Therefore this very low inter-analysers variability has no significant clinical impact. CONCLUSION: This study showed that the analytical system made of Modular P, Cobas Integra, Roche Diagnostics and DakoCytomation reagents can be used for FibroTest-ActiTest-Fibro parameters assessment. Their statistical and clinical variability were acceptable compared to the reference system.

Automation↗