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Biomedical subjects

P Gillery

Publications and source records attributed to P Gillery.

At least 55 records · Page 3Linked to original sources

Changes in serum lipoprotein(a) levels in children with corticosensitive nephrotic syndrome.

Variations in lipoprotein(a) [Lp(a)] levels were evaluated during the course of the nephrotic syndrome in 20 children (17 males, 3 females, aged 2-16 years), to evaluate the use of this parameter in the prognosis and monitoring of the disease. One patient was in relapse, 12 in remission, and 7 alternated between remission and relapse. Results were compared with those obtained in a control population of 100 age-matched children. Lp(a) was measured by a previously described immunonephelometric technique. Serum Lp(a) levels were increased in children with relapsing nephrotic syndrome compared with controls (median value of 419 mg/l vs. 86 mg/l). The median Lp(a) level in patients with nephrotic syndrome in remission was different from controls (270 mg/l under steroid therapy and 163 mg/l without steroid therapy), but remained within the reference range. Of the patients in relapse, 66% had Lp(a) levels above the generally accepted limit for cardiovascular risk of 300 mg/l, compared with 16% of controls, 44% of patients with nephrotic syndrome in remission under steroid therapy, and 18% of patients with nephrotic syndrome in remission without steroid therapy. In 2 patients, Lp(a) was measured repeatedly and was significantly higher during the acute phase of the disease (up to sixfold basal level). Changes in Lp(a) levels correlated with cholesterol levels, but the kinetics and the extent of variations were different. These data suggest that measurement of Lp(a) is useful for monitoring the nephrotic syndrome in children, particularly for detecting complications.

Adolescent↗

Alteration of matrix macromolecule synthesis by fibroblasts from a patient with pachydermoperiostosis.

Pachydermoperiostosis (primary hypertrophic osteoarthropathy) is a very rare genetic disease characterized by pachydermia, periostosis, arthralgia, and finger clubbing. Its pathophysiology is still unclear, but previous studies have reported connective tissue hypertrophy in the skin of these patients. We investigated the synthesis of collagen, fibronectin, and proteoglycans by fibroblasts from affected and unaffected skin from one patient with pachydermoperiostosis and four normal donors. We found that collagen synthesis was largely decreased in fibroblasts from the diseased skin, whereas the synthesis of the small dermatan-sulfate-containing proteoglycan decorin strongly increased. Fibroblasts from the unaffected skin of the patient exhibited syntheses of these macromolecules similar to control fibroblasts from healthy donors. Northern blot and dot blot analyses showed decreased pro alpha 1 (I) collagen in patient's affected and unaffected skin fibroblasts whereas increased decorin mRNA levels were found in fibroblasts from the patient's affected skin. No change in cell proliferation was observed. These data demonstrate an alteration of fibroblast biosynthetic activity in the skin lesions of pachydermoperiostosis, which may be responsible, at least in part, for the patient's phenotype.

Adult↗

Expression of interleukin-4 in scleroderma skin specimens and scleroderma fibroblast cultures. Potential role in fibrosis.

BACKGROUND: Scleroderma (systemic sclerosis) is a fibrotic disease characterized by an uncontrolled tissular accumulation of collagen. Several cytokines have been implicated in the fibroblast activation leading to fibrosis. For instance, we have previously demonstrated that interleukin-4 (IL-4) is a potent activator of collagen synthesis in fibroblast cultures. In this study, using immunocytochemical methods and in situ hybridization, we investigated the expression of IL-4 in normal and scleroderma skin and fibroblast cultures. OBSERVATIONS: Immunocytochemical studies with anti-IL-4 antibody were performed on biopsy specimens from 9 patients with normal skin and 11 patients with scleroderma. The label was intense or strong in 8 of the 11 scleroderma skin specimens, whereas it was negative or faint in 8 of the 9 normal skin specimens (P < .01). In situ hybridization demonstrated a significant increase of the number of IL-4 messenger RNA grains in scleroderma skin compared with normal skin (3.1 +/- 1.5 [mean +/- SD] vs 0.8 +/- 0.7; P < .001). A strongly positive labeling with the anti-IL-4 antibody was found in the 4 scleroderma fibroblast cultures, whereas it was negative in the 5 fibroblast control cultures (P < .05). CONCLUSIONS: Our results demonstrate that IL-4 is strongly expressed in the dermis of a large majority of patients with scleroderma and might be synthesized by scleroderma fibroblasts. We suggest that IL-4 is one of the cytokines implicated in the early steps of the fibrotic process.

Adult↗

Serum lipoprotein (a) concentrations in a population of 819 non-insulin-dependent diabetic patients.

Variations in serum Lp(a) concentrations were studied in a large population of non-insulin-dependent diabetic (NIDDM) patients in relation to long-term complications. Lp(a) concentrations were measured by immunonephelometry in 819 NIDDM subjects and compared with those of 128 controls. Correlations were investigated relative to plasma lipid and glycaemic parameters, body mass index (BMI) and macro- and microvascular complications. Mean absolute and relative variations of Lp(a) concentrations were studied in a subgroup of 245 patients over a one-year period. No significant differences were found between Lp(a) concentrations in NIDDM and control subjects. No relationship was evidenced with macrovascular and microvascular complications or glycaemic control. Mean relative Lp(a) variations were correlated with BMI and absolute and relative variations in triglyceridaemia. These results confirm the absence of any alterations of Lp(a) concentrations in a large cohort of NIDDM patients, either with or without micro- and macrovascular complications, but suggest a particular modulatory role for BMI and serum triglyceride variations.

Adult↗

Stimulation of sulphated glycosaminoglycan and decorin production in adult dermal fibroblasts by recombinant human interleukin-4.

Interleukin-4 (IL-4) is a pleiotropic cytokine expressed by inflammatory cells. Previous work from our laboratory has shown that it stimulates collagen synthesis in fibroblasts. Here we report the effects of recombinant human IL-4 on glycosaminoglycan (GAG) and proteoglycan synthesis in normal dermal fibroblasts from adult donors. IL-4 (10 and 100 units/ml) induced a dose-dependent increase of [3H]glucosamine and [35S]sulphate incorporation into total GAGs. The analysis of the different GAG fractions indicated the enhanced synthesis of dermatan/chondroitin sulphates. IL-4 had no effect on hyaluronan synthesis. The increase of sulphated GAG synthesis was correlated with an increase of proteoglycans in the culture medium. Decorin was identified as the major chondroitin/dermatan sulphate-containing proteoglycan in the culture medium of fibroblasts. Its synthesis was strongly stimulated by IL-4. Both the core-protein synthesis and mRNA expression were enhanced, indicating that the cytokine acted, at least in part, at the pre-translational level. These results indicate that IL-4 is able to modulate not only collagen, but also proteoglycan, production by human fibroblasts. Their implications in physiopathological processes such as wound healing or fibrosis is suggested.

Adult↗

Protein synthesis in collagen lattice-cultured fibroblasts is controlled at the ribosomal level.

Fibroblasts cultivated in three-dimensional lattices exhibit a large decrease of protein synthesis, mainly through transcriptional control. However, no previous work was devoted to a potential ribosomal regulation. We evaluated ribosomal ribonucleic acid (RNA) in monolayer- and collagen lattice-cultured fibroblasts. After one week of culture, total RNA was 60% lower in lattice-cultured fibroblasts than in monolayer-cultured cells. The decrease was identical for 18 S and 28 S rRNA subfractions. The half-life of RNA was much shorter in collagen lattice-cultured fibroblasts than in monolayers. These results suggest that protein synthesis in lattice-cultured fibroblasts is partly regulated at the ribosomal level.

Adult↗

Glycohemoglobin assays evaluated in a large-scale quality-control survey.

We report the results of a national quality-control survey on glycohemoglobin (GHb), monitored in France by the Société Française de Biologie Clinique on behalf of the authority of the "Agence du Médicament." A sample of lyophilized hemolysate was sent to 3109 laboratories. Results were obtained from 2770 laboratories. HbA1C, HbA1, and total GHb were measured by 50%, 24%, and 26% of the participants, respectively. Of these measurements, 79% of the HbA1C results and 76% of the total GHb results, but only 48% of the HbA1 results, were within the +/- 20% limits of the indicated target values. Mean values for the hemolysate ranged from 8% to 11% for HbA1C, from 7% to 12% for HbA1, and from 11% to 13% for total GHb. The interlaboratory CVs ranged from 3% to 20%, according to method used. So, methods used for GHb assay, which are based on various principles, exhibit very different analytical performances. Nonetheless, this large-scale study indicates that some techniques can support transferability of results from laboratory to laboratory.

Autoanalysis↗

[Respective value of glycated hemoglobin and fructosamine assays in the care of diabetes mellitus].

The monitoring of metabolic balance in diabetes mellitus involves the assay of cumulative markers of protein glycation. Glycated hemoglobin, particularly the major component HbA1c, and fructosamine, which reflects glycated plasma protein levels, are the most commonly used parameters. Nevertheless, their utilization is still under discussion with respect to methodologies used, as well as to their respective interest in clinical diabetology. This review shows current opinion concerning the analytical and physiopathological use of these biological indicators.

Chromatography, Ion Exchange↗

[Evaluation of assay methods of glycohemoglobin: reference method and parameter].

In this paper, the characteristics of the analytical procedures allowing the measurement of glycated hemoglobin products are presented. With respect to their respective performances, the authors recommend the specific measurement of HbA1c and the more reliable procedure, high pressure liquid chromatography (HPLC).

Chemistry Techniques, Analytical↗

The effect of different collagens and of proteoglycan on the retraction of collagen lattice.

The effect of various collagens and proteoglycan on the formation and retraction of collagen lattices was tested. The most rapid aggregation of collagen molecules was observed by the use of the least cross-linked collagen fractions (ie pepsin-digested calf skin collagen type I). Lattices formed with more cross-linked collagens (acid soluble collagen-ASC, type III) contracted slowly and less intensively. Unpurified pepsinized cartilage extract containing collagen types II, IX and XI, some glycoproteins and proteoglycans formed lattices rather well. On the contrary, purified collagen type II as well as polymeric collagen (solubilized by denaturing conditions) did not form lattices at all. The lattice formation and retraction was intensified by addition of proteoglycan into the culture medium. The authors suggest that the kinetics of the lattice formation and retraction depends on the amount of collagen cross-links and the concentration of proteoglycan in the culture medium.

Collagen↗

[Glycated hemoglobin: assay methods and problems of standardization].

Glycated hemoglobin can be assayed by methods using different principles, whose performances vary and are difficult to control. The most widely used techniques measure either specifically glycated A1c hemoglobin (HbA1c) or total glycated hemoglobin. They are reviewed in detail in this paper and standardization problems due to the absence of reference materials and methods are discussed. HbA1c seems to be considered the most significant parameter for monitoring diabetic control. Standardization of the assays, calibrated with reference to this parameter, could be performed provided there is strict control of the methods performance.

Biological Assay↗

[Interleukin-4: from B-lymphocyte to fibroblast].

Interleukin-4 (IL-4) has been characterized in 1982 by its ability to induce the proliferation of anti-IgM stimulated B-lymphocytes and IgG1 secretion by these same cells after activation by lipopolysaccharide. It's activity on B-cells explain that it received the former names of B-Cell Growth Factor-1 (BCGF-1) or B-Cell Stimulatory Factor-1 (BSF-1). More recent works have shown that IL-4 exerts pleiotropic effects on a large number of cells. Particularly, it is able to activate the production of extracellular matrix components by fibroblasts. The aim of the present paper is to review main data about this cytokine, with a particular emphasis on its role on the regulation of connective tissue cell activities and on its possible implication in pathophysiological events such as fibrosis, wound healing, and tumor invasion.

Arthritis, Rheumatoid↗

A fluorometric deoxyribonucleic acid assay for tridimensional lattice cultures of fibroblasts.

A fast and sensitive in situ assay of deoxyribonucleic acid in miniaturized lattice cultures of fibroblasts is described. Tridimensional collagen and fibrin lattices prepared in 24-well plates were seeded with 50,000 to 200,000 cells. Cultures were fixed with formaldehyde, rinced with isopropanol, and dried. DNA assay was performed directly in the wells by addition of 3,5-diaminobenzoic acid (DABA) reagent. A calibration curve was prepared with calf thymus DNA. Fluorescence of DNA-DABA was evaluated after 45 min incubation (excitation wavelength 420 nm, emission wavelength 490 nm). The method showed linear results from 0.5 to 10 micrograms DNA and proved sensitive for low cell numbers (50,000 per dish). DNA assay in monolayers and in different types of lattices showed that comparable results were obtained in the different models without interference of the extracellular matrix. This technique is regarded as a costless and efficient tool for evaluating the number of cells in lattices in basal conditions or under pharmacological stimulation.

Aminobenzoates↗

Fibroblastic rheumatism: clinical, histological, immunohistological, ultrastructural and biochemical study of a case.

We report a case of fibroblastic rheumatism (FR). Only eight other cases of this recently described entity have been reported previously. FR is characterized by polyarthralgia and joint stiffness without joint destruction, associated with cutaneous nodules and sclerodactyly. Histology shows an increase in the number of fibroblasts and marked dermal fibrosis. Rheumatological and skin manifestations may improve with corticosteroid therapy. In our patient, immunohistochemical studies of involved and uninvolved skin showed an increase in fibronectin and tenascin deposition. In the dermis, the hyperplastic cells had phenotypic features of muscle, suggesting myofibroblastic differentiation. Ultrastructural study showed an increase in active fibroblastic cells with features of myofibroblasts. A hyperproliferative capacity was observed in fibroblasts cultured from involved skin. Biochemical studies of the production of collagen and non-collagen proteins were performed on these cultured cells, and showed a reduction in collagen and non-collagen protein synthesis by FR fibroblasts. Thus, FR appears to differ from other fibrotic skin diseases such as scleroderma, in that dermal fibrosis may be due predominantly to fibroblast proliferation with myofibroblastic differentiation without any increase in collagen synthesis.

Cell Adhesion Molecules, Neuronal↗

In vivo stimulation of connective tissue accumulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ in rat experimental wounds.

The tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ (GHK-Cu) was first described as a growth factor for differentiated cells. Recent in vitro data showed that it possesses several properties of a potential activator of wound repair. We investigated the effects of GHK-Cu in vivo, using the wound chamber model described previously (Schilling, J.A., W. Joel, and M.T. Shurley, 1959. Surgery [St. Louis]. 46:702-710). Stainless steel wire mesh cylinders were implanted subcutaneously on the back of rats. The animals were divided into groups that received sequential injections into the wound chamber of either saline (control group) or various concentrations of GHK-Cu. At the end of the experiments, rats were killed, wound chambers were collected, and their content was analyzed for dry weight, total proteins, collagen, DNA, elastin, glycosaminoglycans, and specific mRNAs for collagens and TGF beta. In the GHK-Cu-injected wound chambers, a concentration-dependent increase of dry weight, DNA, total protein, collagen, and glycosaminoglycan contents was found. The stimulation of collagen synthesis was twice that of noncollagen proteins. Type I and type III collagen mRNAs were increased but not TGF beta mRNAs. An increase of the relative amount of dermatan sulfate was also found. A control tripeptide, L-glutamyl-L-histidyl-L-proline, had no significant effect. These results demonstrate that GHK-Cu is able to increase extracellular matrix accumulation in wounds in vivo.

Animals↗

Rate nephelometric assay of serum lipoprotein(a).

This nephelometric assay of serum lipoprotein(a) [Lp(a)] is characterized by the use of a specific antibody to generate a high rate of light-scatter formation and the elimination of nonspecific reactions from serum samples by diluting samples in phosphate-buffered saline containing polymer enhancer polyethylene glycol (PEG), 40 g/L, and detergent before the assay. We reacted 100 microL of sixfold-diluted serum in 500 microL of buffer containing PEG with 42 microL of pure polyclonal rabbit antiserum (Dakopatts) directed against human Lp(a) and monitored the reaction by rate nephelometry with the Array Protein System nephelometer (Beckman). The standard curve for the reaction was linear in the Lp(a) range 10-1280 mg/L; antigen excess occurred between 1300 and 1400 mg/L. Calibration was performed with serial dilutions of a standard serum. Precision studies showed within-run and between-run CVs of < 2.1% and 6.9%, respectively. The nephelometric results (y) for 100 serum samples were highly correlated with those obtained by radial immunodiffusion (x) calibrated with the same materials: y = 1.07 (+/- 0.03) x - 16.2 (+/- 8.1) mg/L (r = 0.974, P < 0.001). Storing serum for 3 weeks at 4 degrees C or 3 months at -80 degrees C did not affect the results.

Humans↗