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2.
Allergy ; 70(2): 180-6, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25394543

RESUMO

BACKGROUND: Allergen-specific serum immunoglobulin E detection and quantification have become an important step in allergy diagnosis and follow-up. In line with the current trend of laboratory test accreditation to international standards, we set out to design and assess an accreditation procedure for allergen-specific serum IgE. METHODS: Method validation according to the accreditation procedure under the EN ISO 15189 standard was carried out for allergen-specific immunoglobulin E determination using the fluoroimmunoenzymatic method ImmunoCAP(®) (ThermoFisher). Data were produced by 25 hospital laboratories in France. A total of 29 allergen specificities including mixes, extracts, and molecular allergens were assayed. Allergen-specific serum immunoglobulin E concentrations ranged from 0.1 to 100 kUA /l. RESULTS: Repeatability, reproducibility, and accuracy results fulfilled method validation criteria for automated laboratory tests and proved similar irrespective of the allergen specificity, allergen-specific serum immunoglobulin E concentration, or individual laboratory. CONCLUSION: Allergen-specific serum immunoglobulin E determination with the fluoroimmunoenzymatic method ImmunoCAP(®) is a highly repeatable, reproducible, and accurate method which may be considered as a single analyte assay in view of the EN ISO 15189 accreditation procedure.


Assuntos
Alérgenos/imunologia , Fluorimunoensaio/métodos , Fluorimunoensaio/normas , Hipersensibilidade/diagnóstico , Hipersensibilidade/epidemiologia , Imunoglobulina E/imunologia , Humanos , Hipersensibilidade/imunologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
3.
Clin Exp Allergy ; 44(2): 222-30, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24447084

RESUMO

BACKGROUND: Defining suitable markers to diagnose and monitor allergy and its severity is essential to correctly assign patients for specific immunotherapy. Circulating levels of specific IgE are good markers of sensitization, but not of clinically symptomatic allergy. OBJECTIVE: To quantify circulating interleukin (IL)-4- and IL-13-secreting T cells specific for house dust mite (HDM) in children presenting HDM-allergic asthma associated or not with rhinitis and correlate results with clinical symptoms. METHODS: We analysed 26 children with HDM respiratory disease (allergic rhinitis and asthma) together with six children with non-allergic asthma. Peripheral blood mononuclear cells were stimulated with HDM extract in a 24-h ELISpot assay to quantify the number of HDM-specific IL-4- and IL-13-secreting T cells. Asthma severity and control, and rhinitis severity were scored according to the Global Initiative for Asthma (GINA) and the Allergic Rhinitis and its Impact on Asthma (ARIA) Guidelines. RESULTS: The number of HDM-specific IL-4- and IL-13-secreting T cells was higher in patients with allergic asthma as compared to patients with non-allergic asthma. It varied with the season of blood sampling with two peaks in the fall and early spring. Independently of the season, the number of HDM-specific IL-4-secreting T cells correlated with rhinitis severity (OR = 2; 95% IC:1.1-3.8; P = 0.04). CONCLUSIONS AND CLINICAL RELEVANCE: Allergen-specific IL-4- and IL-13-producing T cells were only detected in HDM-allergic asthmatic children (not in patients with non-allergic asthma). Their numbers correlated with clinical severity of allergic rhinitis.


Assuntos
Antígenos de Dermatophagoides , Asma/sangue , Interleucina-13/sangue , Interleucina-4/sangue , Rinite Alérgica Perene/sangue , Estações do Ano , Linfócitos T/metabolismo , Animais , Asma/imunologia , Asma/patologia , Criança , Estudos Transversais , Humanos , Interleucina-13/imunologia , Interleucina-4/imunologia , Contagem de Linfócitos , Pyroglyphidae , Rinite Alérgica , Rinite Alérgica Perene/imunologia , Rinite Alérgica Perene/patologia , Índice de Gravidade de Doença
4.
Allergy ; 69(11): 1473-80, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25041079

RESUMO

BACKGROUND: Cow's milk allergy (CMA) is a frequent food allergy in young children. The oral food challenge is the gold standard for diagnosis, and there is currently no reliable biological test. Our aim was to evaluate the diagnostic potential of a functional assay quantifying allergen-specific Th2 cells in CMA children. METHODS: A total of 29 children aged 2.8-10.5 years underwent a double-blind, placebo-controlled food challenge (DBPCFC) to cow's milk. Blood was collected before performing the DBPCFC, and peripheral mononuclear cells were cultured in an 18-h ELISpot assay with casein, α-lactalbumin, or ß-lactoglobulin. Numbers of antigen-specific IL-4- and IL-13-secreting lymphocytes and serum-specific IgE, IgG4, and total IgE levels were assessed. Receiver operating characteristic (ROC) curves were generated. RESULTS: A total of 17 (59%) children reacted to cow's milk and were therefore considered as allergic to cow's milk (CMA). The mean number of casein-specific IL-4- and IL-13-secreting T cells was higher in CMA than in non-CMA children (P = 0.009, 0.004, respectively). Moreover, it was inversely correlated with the cumulative dose of cow's milk tolerated (P = 0.003, 0.0009, respectively). ROC curve of combined IL-4 and IL-13 analysis showed an area under the curve of 0.98 (95% CI 0.90-1.06). For a cutoff of 10 IL-4- and 12 IL-13-secreting T cells, sensitivity and negative predictive value were 100%. CONCLUSIONS: Enumeration of casein-specific IL-4- and IL-13-secreting T cells appears a promising tool to improve diagnosis and, if confirmed in larger studies, could permit less frequent use of the oral food challenge.


Assuntos
Caseínas/imunologia , Interleucina-13/metabolismo , Interleucina-4/metabolismo , Hipersensibilidade a Leite/imunologia , Hipersensibilidade a Leite/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Animais , Bovinos , Criança , Pré-Escolar , ELISPOT/métodos , ELISPOT/normas , Feminino , Humanos , Tolerância Imunológica , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Hipersensibilidade a Leite/diagnóstico , Estudos Prospectivos , Curva ROC , Reprodutibilidade dos Testes , Especificidade do Receptor de Antígeno de Linfócitos T/imunologia
6.
J Med Genet ; 46(7): 490-4, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19443464

RESUMO

BACKGROUND: Mutations in the surfactant protein C gene (SFTPC) have been recently associated with the development of diffuse lung disease, particularly sporadic and familial interstitial lung disease (ILD). OBJECTIVE: We have investigated the prevalence and the spectrum of SFTPC mutations in a large cohort of infants and children with diffuse lung disease and suspected with surfactant dysfunction. METHOD AND RESULTS: 121 children were first screened for the common SFTPC mutation, p.Ile73Thr (I73T). Ten unrelated patients were shown to carry this mutation. The I73T mutation was inherited in six cases, and appeared de novo in four. The 111 patients without the I73T mutation were screened for the entire coding sequence of SFTPC. Of these, eight (seven unrelated) subjects were shown to carry a novel mutant allele of SFTPC. All these seven new mutations are located in the BRICHOS domain except the p.Val39Ala (V39A) mutation, which is in the surfactant protein C (SP-C) mature peptide. CONCLUSIONS: Our results confirm that SFTPC mutations are a frequent cause of diffuse lung disease, and that I73T is the most frequent SFTPC mutation associated with diffuse lung disease.


Assuntos
Pneumopatias/genética , Mutação , Proteína C Associada a Surfactante Pulmonar/genética , Criança , Pré-Escolar , Estudos de Coortes , Análise Mutacional de DNA , Feminino , Humanos , Lactente , Masculino , Linhagem
7.
Eur J Neurol ; 16(7): 832-7, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19473356

RESUMO

BACKGROUND AND PURPOSE: The influence of apolipoprotein E (ApoE) polymorphism on clinical severity of multiple sclerosis (MS) is still controversial. Cigarette smoking has been suggested to influence the progression of disability in these patients. In this study, we aimed to investigate whether an interaction of smoking with the ApoE polymorphism influences the progression of disability in MS patients. METHODS: Smoking history from 205 female patients with MS was obtained. Clinical data collected include age at onset, disease duration, annual relapse rate, the Expanded Disability Status Scale (EDSS) and the Multiple Sclerosis Severity Score (MSSS). ApoE polymorphism was examined in all patients and stratified according to smoking status and associations with the clinical data investigated. RESULTS: There were no significant associations between cigarette smoking and any of the clinical characteristics in the whole group of patients. In women carrying the ApoE E4 isoform, smokers had a lower EDSS (P = 0.033) and MSSS (P = 0.023) in comparison with non-smokers. CONCLUSION: Our data suggest that in women with MS carrying the ApoE E4 isoform, cigarette smoking may have a protective influence on disease progression and accumulation of disability. These findings need to be confirmed by future large longitudinal studies.


Assuntos
Predisposição Genética para Doença , Esclerose Múltipla/genética , Esclerose Múltipla/fisiopatologia , Polimorfismo Genético/genética , Fumar/genética , Adulto , Apolipoproteínas E/genética , Avaliação da Deficiência , Progressão da Doença , Feminino , Humanos , Pessoa de Meia-Idade , Índice de Gravidade de Doença
8.
Int J Pediatr Otorhinolaryngol ; 70(4): 689-96, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16226319

RESUMO

OBJECTIVE: The purpose of the study is to present the results of cochlear implantation in case of deafness involving mutations in the OTOF gene. This form of deafness is characterized by the presence of transient evoked otoacoustic emissions (TEOAE). In cases of profound deafness with preserved TEOAE, two main etiologies should be considered: either an auditory neuropathy (a retrocochlear lesion) or an endocochlear lesion. It is essential to differentiate these two entities with regards to therapy and screening. PATIENTS: We report two children who presented with profound prelingual deafness, confirmed by the absence of detectable responses to auditory evoked potentials (AEP), associated with the presence of bilateral TEOAE. Genetic testing revealed mutations in OTOF, confirming DFNB9 deafness. Both patients have been successfully implanted (with a follow-up of 18 and 36 months, respectively). MAIN OUTCOME MEASURES: Clinical (oral production, closed and open-set words and sentences list, meaningful auditory integration scale), audiometric evaluation (TEOAE, AEP) before and after implantation, and neural response telemetry (NRT). RESULTS: Both patients present a good quality of clinical responses and electrophysiological tests after implantation, indicating satisfactory functioning of the auditory nerve. This confirms the endocochlear origin of DFNB9 and suggests that these mutations in OTOF lead to functional alteration of inner hair cells. CONCLUSION: In the absence of a context of neurological syndrome, the combination of absent AEP and positive TEOAE should lead to a genetic screening for mutations in OTOF, in order to undertake the appropriate management.


Assuntos
Implante Coclear , DNA/análise , Potenciais Evocados Auditivos do Tronco Encefálico/fisiologia , Perda Auditiva Neurossensorial/genética , Perda Auditiva Neurossensorial/cirurgia , Proteínas de Membrana/genética , Mutação , Emissões Otoacústicas Espontâneas , Audiometria , Pré-Escolar , Análise Mutacional de DNA , Feminino , Perda Auditiva Neurossensorial/congênito , Humanos , Lactente , Fenótipo
9.
J Leukoc Biol ; 50(1): 1-9, 1991 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-1647432

RESUMO

In this work, the in vitro effects of very-low-density lipoproteins (VLDL) on human polymorphonuclear leukocyte (PMN) oxidative metabolism and migration were studied. VLDL stimulated PMN superoxide generation in absence of other stimulating agents. The effect of VLDL from normotriglyceridemic subjects was more marked than with VLDL from hypertriglyceridemic subjects. VLDL reduced in a dose-dependent manner the luminol-dependent chemiluminescence of PMN stimulated by phorbol myristate acetate (PMA) and, to a lesser degree, by opsonized zymosan. This effect was observed with VLDL concentrations found in healthy and hypertriglyceridemic patients. Superoxide anion generation was also reduced by preincubation of PMN with VLDL before stimulation with PMA but not opsonized zymosan. VLDL were not cytotoxic for PMN. The above effects appear to be an intrinsic property of VLDL and might lead to reduced PMN-mediated non-specific host defences in hypertriglyceridemic subjects.


Assuntos
Lipoproteínas VLDL/farmacologia , Neutrófilos/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Humanos , Técnicas In Vitro , Lipossomos/farmacologia , Medições Luminescentes , Neutrófilos/metabolismo , Oxirredução , Superóxidos/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Zimosan/farmacologia
10.
Ann Biol Clin (Paris) ; 63(5): 443-55, 2005.
Artigo em Francês | MEDLINE | ID: mdl-16230278

RESUMO

Arginine, a semi-essential amino acid, plays a major nutritional and metabolic role. In particular, arginine is the precursor of nitric oxide which is involved in the endothelial function. Several factors, such as hypercholesterolemia, diabetes, ageing and hypertension are established risk factors for atherosclerosis, in particular by decreasing the availability of nitric oxide. Thus, endothelial nitric oxide synthase has a pivotal role against atherosclerosis. A suitable amount of cofactor and a sufficient intake of arginine have been shown to modulate nitric oxide-induced vasodilatation: despite the fact that the intracellular concentration of arginine is well above the Km of endothelial nitric oxide synthase, an arginine supplemented-diet is effective in increasing the production of nitric oxide. Several mechanisms have been proposed to explain this "arginine paradox": co-localization of the arginine transporter with endothelial nitric oxide synthase, intracellular arginine regeneration from citrulline, balance between endothelial arginase and nitric oxide synthase. Statins which are HMG-CoA reductase inhibitors inhibit the synthesis of mevalonate, and thus that of cholesterol. In addition, statins increase the stabilization of endothelial nitric oxide synthase mRNA. The co-operation between cholesterol synthesis and the upregulation of caveolin-1 on the one hand, and the activation of endothelial nitric oxide synthase on the other hand, is very tight. A depletion of cholesterol in the caveolae induces a decrease in caveolin-1 at the cell surface allowing NOS activation. Thus statins improve nitric oxide production and vasodilatation. In a recent work in the hypercholesterolemic Watanabe rabbit, we have demonstrated that the combination of arginine with a statin, namely atorvastatin, significantly hinders the spreading of atherosclerotic plaques as compared with monotherapies. Such association of a nutriment and a drug open a new area of therapeutic strategy.


Assuntos
Anticolesterolemiantes/uso terapêutico , Arginina/metabolismo , Arginina/uso terapêutico , Aterosclerose/fisiopatologia , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Óxido Nítrico/fisiologia , Envelhecimento , Arginina/administração & dosagem , Aterosclerose/epidemiologia , Suplementos Nutricionais , Humanos , Hipercolesterolemia/complicações , Hipertensão/complicações , Modelos Biológicos , Óxido Nítrico Sintase Tipo III/genética , RNA Mensageiro/genética , Fatores de Risco
11.
Ann Biol Clin (Paris) ; 63(1): 27-41, 2005.
Artigo em Francês | MEDLINE | ID: mdl-15689310

RESUMO

International guidelines emphasize the importance of LDL cholesterol (LDL-C) assay in the care and follow-up of patients with cardiovascular risk. Most studies and common practice use Friedewald's formula for LDL-C calculation. The accuracy of the result depends closely on the precision of the input parameters (total cholesterol, triglycerides (TG) and HDL cholesterol), and discrepancies between calculated LDL-C and measurement by reference methods appear when TG exceed 4.5 mmol/L, or in the presence of abnormal lipoproteins. These restrictions and uncertainties in calculations have prompted the recent development of direct and homogeneous methods that fit all analyzers. A multicenter evaluation of four direct assays of LDL-C (Daiichi, Denka Seiken, Kyowa, Wako) was carried out on 45 serum samples (TG below 3.1 mmol/L) in eight laboratories using different analyzers. For three methods (Daiichi, Kyowa, Wako), the interlaboratory reproducibility was markedly improved relative to that of calculation. A strong correlation was found for all new methods when compared with a beta-quantification assay. Average bias in Denka Seiken assays was greater than Kyowa's and Daiichi's (although less dispersed for the latter) and for Wako all bias were positive. The relationship between bias variations and the lipid parameters of the samples was studied. Three methods, Daiichi, Kyowa and Wako, revealed a significant positive correlation between bias and serum VLDL-C/TG ratio, clearly indicating that cholesterol enrichment of VLDL was a source of variability in these assays. Specificity of the four methods was tested in situation of dyslipidemia by spiking isolated lipoproteins (chylomicrons, VLDL and HDL). This experiment revealed differences in behavior, most evidently upon addition of VLDL. No method was truly specific, but up to 8 mmol/L of TG the variations were acceptable. In the presence of type III hyperlipoproteinemia, however, only the Denka Seiken method was reliable. Linearity up to 20 mmol/L (Daiichi, Denka Seiken) or 14 mmol/L (Kyowa, Wako) of LDL-C allows these tests to be used in main routine cases. New direct assays are an obvious technological advance in terms of analytical performance and conveniency. Their use for the diagnosis and follow-up of hyperlipidemic patients offers an alternative that overcomes the limitations of the Friedewald calculation.


Assuntos
LDL-Colesterol/sangue , Análise Química do Sangue/métodos , Colesterol/sangue , Humanos , Hiperlipoproteinemias/sangue , Laboratórios , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Triglicerídeos/sangue
12.
Artigo em Inglês | MEDLINE | ID: mdl-26319306

RESUMO

The main objective of this work was to evaluate a comprehensive two-dimensional gas chromatographic (GCxGC) coupled to quadrupole mass spectrometry (qMS) method in the field of biomarker candidates' discovery. To this purpose we developed a GCxGC-qMS method suitable for the separation of organic acids and other classes of compounds with silylable polar hydrogen such as sugars, amino-acids, and vitamins. As compared to those obtained by a widely used 1D-GC method, the urinary chromatographic profiles performed by the proposed 2D-GC method exhibit higher resolution and sensitivity, leading to the detection of up to 92 additional compounds in some urine samples including some well-known biomarkers. In order to validate the proposed method we focused on three metabolites of interest with various functional groups and polarities including CH3-malonic acid (MMA: biomarker of methylmalonic acidemia), 3-hydroxy-3-methyl-glutaric acid (3-OHMGA: biomarker of 3-hydroxy-3-methylglutaric acidemia), and phenylpiruvic acid (PhPA: marker of phenylketonuria). While these three metabolites can be considered as representative of organic acids classically determined by 1D-GC, they cannot be representative of new detected metabolites. Thus, we also focused on quinolic acid (QUIN), taken as an example of biomarker not detected at basal levels with the classical 1D GC-qMS method. In order to obtain sufficient recoveries for all tested compounds, we developed a sample preparation protocol including a step of urea removal followed by two extraction steps using two solvents of different polarity and selectivity. Recoveries with the proposed method reached more than 80% for all targeted compounds and the linearity was satisfactory up to 50µmol/L. The CVs of the within-run and within-laboratory precisions were less than 8% for all tested compounds. The limits of quantification (LOQs) were 0.6µmol/L for MMA, 0.4µmol/L for 3-OHMGA, 0.7µmol/L for PhPA, and 1µmol/L for QUIN. The LOQs of these metabolites obtained by a classical GC-MS method under the same chromatographic conditions were 5µmol/L for MMA, 4µmol/L for 3-OHMGA, 6µmol/L for PhPA while QUIN was below the limit of detection. As compared to 1D-GC, these results highlight the enhanced detectability of urine metabolites by the 2D-GC technique. Our results also show that for each new detected compound it is necessary to develop and validate an appropriate sample preparation procedure.


Assuntos
Cromatografia Gasosa/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Ácidos Quinolínicos/urina , Criança , Humanos
13.
Am J Med Genet ; 54(3): 286-8, 1994 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-7810590

RESUMO

Apolipoprotein E, type epsilon 4 allele (ApoE epsilon 4), is associated with late-onset sporadic Alzheimer's disease (AD) in French patients. The association is highly significant (0.45 AD versus 0.12 controls for epsilon 4 allele frequencies). These data support the involvement of ApoE epsilon 4 allele as a very important risk factor for the clinical expression of AD.


Assuntos
Alelos , Doença de Alzheimer/genética , Apolipoproteínas E/genética , Idoso , Sequência de Bases , DNA/análise , Genótipo , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Fatores de Tempo
14.
Am J Med Genet ; 102(3): 219-26, 2001 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-11484197

RESUMO

Pierre Robin sequence (PRS) consists of the nonrandom association of micrognathia, cleft palate (CP), and glossoptosis. It also includes respiratory and feeding difficulties that appear to be neurogenic rather than mechanical in causation. Genetic determinants are thought to underlie this functional and morphological entity, based on the existence of Mendelian syndromes with PRS, and the rare observations of familial nonsyndromic PRS, in which some of the affected individuals have isolated CP. We report the association of PRS with deletion 2q32.3-q33.2 due to an unbalanced reciprocal translocation 46,XX, t(2;21), del 2(q32.3q33.2), and we refine the deletion interval with regard to YAC probes and polymorphic DNA markers. The deletion was shown to be flanked by D2S369 (telomeric) and D2S315 (centromeric), thus it maps to a recently determined chromosomal region known to be nonrandomly associated with CP. This observation supports the hypothesis for the genetic bases of nonsyndromic PRS, strengthens its possible genetic association with isolated CP, and provides a candidate PRS locus, in chromosomal region 2q32.3-q33.2.


Assuntos
Deleção Cromossômica , Cromossomos Humanos Par 2/genética , Síndrome de Pierre Robin/genética , Bandeamento Cromossômico , Mapeamento Cromossômico , Feminino , Humanos , Hibridização in Situ Fluorescente , Lactente , Repetições de Microssatélites , Síndrome de Pierre Robin/patologia
15.
Am J Med Genet ; 104(1): 86-92, 2001 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-11746036

RESUMO

Cleft lip with or without cleft palate (CL/P) is one of the most common congenital malformations in humans occurring with a birth prevalence of approximately 1:1,000. CL/P may be part of a defined syndrome, sequence or association, although most individual or familial cases present as an isolated (nonsyndromic) malformation (NSCL/P). Inheritance is generally regarded as multigenic although, in some families, NSCL/P seemingly segregates as a monogenic trait. On the other hand, van der Woude syndrome (vWS) is a rare autosomal dominant with cardinal features of lower-lip pits (LLP) and CL/P or cleft palate (alone). Since none of these traits is present in all mutation carriers, some individual or familial vWS cases, especially those lacking LLP, are indiscernible from NSCL/P, raising the question whether allelic variation at the vWS locus could underlie NSCL/P. This question was addressed using parametric linkage (LOD score) analysis in 21 multiplex NSCL/P families based on a tightly linked microsatellite marker (D1S3753), and nonparametric analysis using the transmission/disequilibrium test (GTDT) in 106 NSCL/P triads and selecting markers D1S205, D1S491, and D1S3753. No evidence for linkage of NSCL/P to vWS was found on the 21 families using the LOD score approach. In contrast, TDT yielded a significant P value of 0.04 for D1S205, supporting involvement of vWS in NSCL/P in a complex, modifying/polygenic manner rather than as a monogenic/major disease locus.


Assuntos
Fenda Labial/genética , Fissura Palatina/genética , Alelos , Feminino , Genótipo , Heterozigoto , Humanos , Anormalidades Maxilomandibulares , Desequilíbrio de Ligação/genética , Escore Lod , Masculino , Repetições de Microssatélites/genética , Linhagem
16.
Diabetes Metab ; 25(4): 350-5, 1999 Sep.
Artigo em Francês | MEDLINE | ID: mdl-10566127

RESUMO

Analytical evaluation three models of blood glucose meters (Glucometer IV (Bayer), Glucotrend (Roche Diagnostics) and One touch (Life Scan, Johnson & Johnson)) were compared within a central laboratory to an analyzer (Hitachi 747, Boehringer-Mannheim, Grenoble, France) and to a photometric device (HemoCue B-glucose). Each glucose meters met the criteria of within-run precision defined by the Association Française d'Etudes du Diabète et des Maladies Métaboliques (ALFEDIAM). Furthermore, HemoCue B-glucose met criteria defined by the French Society of Biological Chemistry (SFBC). In the study realized within laboratory, 28 to 66% of glucose tests realized with meters differed by more than 10% from plasma values and 43% of glucose assays realized with different meters differed by more than 10% from HemoCue B-glucose value. Within clinical departments, 46 glucose meters from 5 different models were evaluated. Answers to a questionnaire stored in HemoCue B-glucose underlined the weak development of glucose meters quality assurance. Within clinical departments, 65% of glucose values realized with meters differed by more than 10% from reference values. Development in clinical departments of glucose meters quality assurance programs can be monitored by HemoCue B-glucose, and must allowed improvement of glucose meter quality.


Assuntos
Análise Química do Sangue/instrumentação , Glicemia/análise , Sistemas Automatizados de Assistência Junto ao Leito/normas , Estudos de Avaliação como Assunto , Humanos , Garantia da Qualidade dos Cuidados de Saúde , Reprodutibilidade dos Testes
17.
Clin Chim Acta ; 244(1): 1-15, 1996 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-8919198

RESUMO

We have characterized changes in lipoproteins from cholestatic individuals and reproduced them by incubating lipoproteins from healthy individuals with cholic acid. The cholestatic patients showed an increase in low density lipoprotein (LDL) (>85%), with a smaller proportion of esterified cholesterol, and a fall in high density lipoprotein (HDL) (<10%), with a larger proportion of phospholipids. The protein composition of cholestatic HDL1 was characterized by a smaller proportion of apo A (I, II) and a prominent apo E fraction (39% vs. 9%). These changes involved an increase in degree of molecular packing (order) of HDL1. The addition of cholic acid to serum from healthy individuals altered the lipoprotein distribution, with an increase in LDL, the disappearance of HDL2 and HDL3 and the appearance of HDL1. These HDL1 were characterized by increased phospholipid and reduced apo AI fractions. They also showed a lower density and appeared as spherical particles in contrast to cholestatic HDL. Incubation of healthy HDL with cholic acid in vitro reproduces some of the alteration observed in cholestatic HDL.


Assuntos
Colestase Extra-Hepática/sangue , Colestase Intra-Hepática/sangue , Lipoproteínas/sangue , Fenômenos Químicos , Físico-Química , Ácido Cólico , Ácidos Cólicos/farmacologia , Polarização de Fluorescência , Humanos , Imunodifusão , Lipoproteínas/química , Lipoproteínas/efeitos dos fármacos , Lipoproteínas HDL/química
18.
Clin Chim Acta ; 277(2): 141-52, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9853697

RESUMO

Delayed postprandial clearance of triglyceride-rich lipoproteins (TGRL) could induce a decrease of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) vitamin E content through its transfer into TGRL. We thus studied lipoprotein vitamin E content during postprandial hypertriglyceridemia induced by a high fat meal without vitamin E supplement. Venous blood was drawn from five healthy male volunteers following a 12 h fast at (t0) then 2 h (t2), 4 h (t4), 6 h (t6) and 8 h (t8) after a 80 g fat meal. In plasma, only TG significantly varied during the postprandial period with a large interindividual variability. Mean composition of lipoproteins in terms of mass was not significantly modified. The amount of vitamin E significantly increased in TGRL and decreased in LDL plus HDL at t4 and t6 relative to t0. Vitamin E content of TGRL and LDL but not of HDL decreased significantly at t4. The mean decrease was 20% (range 5%-54%) for the LDL. LDL- and HDL vitamin E content correlated inversely with plasma TGRL levels. Our data suggest that LDL from subjects with delayed chylomicron clearance could be less protected against oxidation.


Assuntos
Hipertrigliceridemia/sangue , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Período Pós-Prandial , Vitamina E/metabolismo , Gorduras na Dieta/administração & dosagem , Eritrócitos/metabolismo , Humanos , Lipoproteínas HDL/química , Lipoproteínas LDL/química , Masculino , Valores de Referência
19.
Clin Chim Acta ; 224(1): 65-71, 1994 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-8174279

RESUMO

The biological functions of alpha-1 acid glycoprotein (AGP) are poorly understood but appear to depend on glycan microheterogeneity. Variations of AGP glycan structure (in terms of concanavalin A (ConA) reactivity) have been observed during the inflammatory process. We studied these modifications in AGP from patients with chronic renal impairment and investigated the effects of AGP microheterogeneity on healthy polymorphonuclear leukocyte (PMN) chemotaxis and oxidative metabolism. AGP was extracted by a two-step procedure from sera from ten patients with various degrees of renal impairment, selected according to AGP glycan heterogeneity determined by crossed immunoaffinity electrophoresis with ConA. AGP (0.5 g/l) significantly inhibited the chemotactic response of PMN to formyl-methionyl-leucyl-phenylalanine (10(-7) mol/l) and complement fraction C5a, regardless of ConA reactivity. AGP also inhibited superoxide anion generation in response to phorbol myristate acetate (10(-7) mol/l). After stimulation by opsonized zymosan (1 g/l), the effect of AGP appeared to depend on its glycan structure (r = 0.70, P < 0.05), decreasing with ConA non-reactivity. These data suggest that AGP can down-regulate neutrophil responsiveness, an effect that depends in part on its glycan microheterogeneity. Alterations of AGP microheterogeneity in various pathological states, particularly renal failure, may be related to the inflammatory process.


Assuntos
Neutrófilos/efeitos dos fármacos , Orosomucoide/farmacologia , Polissacarídeos/farmacologia , Adulto , Quimiotaxia de Leucócito/efeitos dos fármacos , Complemento C5a/farmacologia , Concanavalina A/farmacologia , Humanos , Técnicas In Vitro , Nefropatias/metabolismo , Pessoa de Meia-Idade , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Orosomucoide/química , Oxirredução , Polissacarídeos/química , Superóxidos/metabolismo
20.
Clin Nephrol ; 54(3): 210-7, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11020019

RESUMO

AIMS: Patients with chronic renal failure treated by hemodialysis develop lipoprotein abnormalities that may contribute to their increased risk ofatherosclerosis. This study shows lipid parameter follow-up procedure according to the type of dialysis membrane in an unselected population of 33 hemodialysis patients. PATIENTS AND METHODS: The study included 33 patients with end-stage renal disease and 110 healthy blood bank donors of Tenon Hospital. Cholesterol and triglycerides were determined by enzymatic methods, apoA-I, apoB by immunoturbidimetry and Lp(a) by immunonephelemetry. Apo(a) phenotyping was performed by agarose gel electrophoresis followed by immunoblotting. Patients and controls subjects were estimated by Student's t- and chi2-tests. RESULTS: Patients dialyzed with low-flux membranes had Lp(a) concentrations higher than patients dialyzed with high-flux membranes. Patients dialyzed with polyacrylonitrile membranes (AN 69) had an apoA-I concentration significantly lower than patients dialyzed with hemophane or polysulfone membranes. We also confirmed some of the lipid abnormalities and high Lp(a) concentrations in ESRD patients. CONCLUSION: These results may contribute to a more rational choice of the dialysis membrane in hemodialysis patients.


Assuntos
Falência Renal Crônica/sangue , Falência Renal Crônica/terapia , Lipoproteína(a)/sangue , Diálise Renal , Adulto , Idoso , Feminino , Seguimentos , Humanos , Masculino , Membranas Artificiais , Pessoa de Meia-Idade , Fatores de Tempo
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