[Diagnosis of adult-onset Still's disease: contribution of biology]. / Apport du laboratoire dans le diagnostic de la maladie de Still.

We report one case of adult-onset Still's disease. The diagnosis of this disease currently requires application of Yamaguchi's criteria and then exclusion of infectious, haematological, autoimmune and neoplasic disorders. The place of biology is enhanced with the use of new markers like ferritin and glycosylated ferritin. Low percentage of glycosylated ferritin (<20%) in presence of very high level of ferritin, unexplained prolonged fever with leukocytosis can help to diagnosis. However, glycosylated ferritin is not yet of in routine use.

[Study of present possibilities of interlaboratory standardisation of the prothrombin time (author's transl)]. / Etude sur les possibilités actuelles de standardisation interlaboratoire du temps de Quick.

The graphs or tables proposed by the manufacturer of reagents, the Thivolle straight lines suggested by Raby or, since 1977, Loeliger's procedure, are three possibilities offered to biologists to transform the prothrombin time into prothrombin concentrations expressed as a percentage. We have studied the possibility of interlaboratory standardisation in the case of patients treated with coumarin derivatives: firstly with the same thromboplastin under various conditions of use; secondly with five different thromboplastins under identical conditions (choosing arbitrarily one of the reagents as reference). Loeliger's procedure is the one which permits the best standardisation, but its practical application seems difficult. It seems to us that before carrying out this standardisation between laboratories, one should indicate, with the result found for the patient, the area of therapeutic activity of the thromboplastin used, as suggested by Ducker and Marbet in 1977 (these authors determined this area of therapeutic activity for 13 different reagents used in Switzerland).

[Hereditary bisalbuminemia. Study of a new familial case in France (author's transl)]. / Bisalbuminémie héréditaire. Etude d'un nouveau cas familial français.

A case of bisalbuminemia of slow type was discovered in a 41 year old woman with obesity, diabetes mellitus and hypertension. This abnormality was discovered in six other members of the family and was thus hereditary. After a description of the laboratory tests used, the various forms of bisalbuminemia are described together with the circumstances of onset of the acquired forms, the biochemical and metabolic characteristics of the hereditary forms and their mode of transmission. Bisalbumin is present in all races. The mother of our case was of Italian origin, which permitted us to classify this case with others of Italian origin.

[Measurement of peroxidase activity with orthophenylenediamine as chromogen in immunoenzyme methods]. / Mesure de l'activité peroxydasique avec l'orthophénylènediamine comme chromogène dans les méthodes enzymo-immunologiques.

The authors studied orthophenylenediamine in order to determine the optimal conditions for its use in "sandwich" type of enzymo-immunology. They studied the kinetics of the reaction and the stopping of the enzymatic reaction in the presence of peroxidase-labeled antibody. Studies of the dissolution of orthophenylenediamine and the stability of the solutions obtained show that a solution of orthophenylenediamine can be stored at ambiant temperature and in darkness for 8 hours. Depending on the degree of sensitivity required, the concentration of orthophenylenediamine can vary between 10 to 30 mmol/l with 10 mmol/l of H2O2. Stopping the reaction with acid greatly increases the values of absorbance by 1.5 to 3 times, depending on the type of acid used. The tests demonstrated that the sensitivity and repeatability are better than can be obtained with 2,2' azino-di(3-ethylbenzthiazoline-6-sulfonic) acid. On the basis of these experiments, orthophenylenediamine was used in the assay of ferritin and total IgE: the increased sensitivity meant that the amount of antigen introduced into the reaction could be reduced to 1/5 of the amount usually required.

[Erythrocyte ferritin]. / La ferritine érythrocytaire.

Red cell ferritin is a residue of erythroblast ferritin. It reflects the balance between the iron supply to the erythroid marrow and the need for haemoglobin synthesis. Erythrocyte ferritin can be measured in haemolysates after discarding plasma and leucocytes by different methods. The decrease in erythrocyte ferritin content indicates manifest iron deficiency anaemia. Ferritin levels do not appear to be influenced by inflammation, infection, tissue necrosis or tumors and may be a reliable indicator of iron status in inflammatory diseases. Erythrocyte ferritin is markedly increased in patients with iron overload allowing early diagnosis of hereditary haemochromatosis and the monitoring of phlebotomy therapy. Finally, in some pathologies erythrocyte assay is a more reliable indicator of the iron status than serum ferritin.

Modification of the selected method for the determination of serum iron. Substitution of bathophenanthroline by ferene S.

The previously selected method (1977) for the measurement of iron in human serum has been modified by the working group on Iron and Iron-Transport Proteins (Sociéte Française de Biologie Clinique). Ferene S is used as the chromogen, it is more sensitive and cheaper than bathophenanthroline. The sample size has been reduced from 2 to 0.5 ml. No interference could be shown with high concentrations of bilirubin, copper or hemoglobin.

[Hemodialysis and ferritinemia]. / Hémodialyse et ferritinémie.

Iron-deficiency is a common phenomenon in chronic renal diseases and haemodialysis patients, a treatment with iron or transfusions is always provided in an early preventive way; yet, an overload may appear. Serum ferritin, in spite of analytic variability, remains at the present time a good witness to appreciate patients' iron stores. Authors report the results obtained with four commercial reagents in healthy population and in haemodialysed children or adults. The comparison of results for this parameter shows that haemodialysed sera, with or without treatment, have the same behaviour with reagents as those of healthy subjects.

[Reference values of erythrocyte ferritin in children and adults]. / Valeurs de référence de la ferritine érythrocytaire chez l'enfant et l'adulte.

The erythrocyte ferritin content was measured in 183 healthy subjects in age from 4 to 68 years; 80 were male and 103 were female. In children between the ages of 4 and 12 years there is no significant difference in the mean value between boys and girls. In females the erythrocyte ferritin concentration is independent of age. After 12 years of age the erythrocyte ferritin content is higher in men. Reference intervals were determined by the two quantiles 0.05 and 0.95. The reference interval is 3-24 age by cell in boys between 4 and 12 and females between 4 and 63; the reference interval is 5-38 age by cell in males between 13 and 68 years of age.

Comparison of six serum ferritin immunoassays and isoferritin spectrotypes in malignancies.

Serum ferritin was measured by six enzyme immunoassays in specimens from patients with digestive cancers (n = 30) and hematologic malignancies (n = 33). Most mean comparisons show significant differences in both groups of patients. In digestive cancers correlations between any two methods are very satisfactory (r > 0.99) but a proportional bias is often observed. In hematologic malignancies, correlations are bad (r < 0.80 in 8 out of 15 correlations) because of many discrepant values. Isoelectric focusing separation of isoferritins was performed in most specimens and the pattern of each serum was compared to the between kit CV. We conclude that an 'acid' spectrotype increases between-kit analytical variability. We try to explain the results taking into account the nature of the immunological systems and the cross-reactions with tissular isoferritins. In conclusion, our results indicate that large differences may be observed in sera from hematologic malignancies (leukemias, lymphomas ... ) We recommend that monitoring be achieved by the same method of measurement.

[Multicenter study of sialic acid deficient transferrin determined by two chromatographic techniques]. / Etude multicentrique du dosage de la transferrine déficiente en acide sialique par deux techniques chromatographiques.

Serum carbohydrate-deficient-transferrin (CDT) was measured by a micro anion-exchange chromatography/enzyme immunoassay. Results obtained on 245 sera analyzed in four laboratories were compared. Moreover, one laboratory used a commercial kit with ready-to-use microcolumns and a radioimmunoassay for measuring eluted CDT. Imprecision was judged to be satisfactory. Within-assay coefficients of variation ranged from 5 to 10%, between-assay coefficients of variation ranged from 9 to 18%. Between-laboratory results were compared for 110 sera from control subjects (daily alcohol intake < 40 g), for 57 sera from chronic ethylic subjects and for 78 sera from patients suffering from non-alcoholic liver diseases. There was a large between-laboratory variation, suggesting that the method is difficult to standardize and that results are not transferable. Results of enzyme and radioimmunoassays were compared on 325 sera. The best correlation was obtained in the groups of ethylic subjects and those with non-alcoholic hepatic diseases. Finally the performance of the CDT-test was evaluated by calculating sensitivity and specificity. With both methods specificity was very high (> 85%) but sensitivity was poor (< 50%).

[Multicenter evaluation of thr Abbott glycosylated hemoglobin assay on IMx]. / Evaluation multicentrique de test Abbott hémoglobine glyquée sur IMx.

The Abbott IMx glycated hemoglobin assay was evaluated in a multicentre study. This method utilizes boronate affinity chromatography, and ion-capture technology. This assay determines both total glycohemoglobin (% GHb) and percentage of hemoglobin Alc (% HbAlc). The precision of the assay was evaluated: the intra-assay and interassay coefficients of variation were judged to be satisfactory (< 6.5%). We determined the accuracy of the assay by comparison with a reference HPLC assay for 603 specimens; coefficients of correlation were between 0.88 and 0.96. We studied the interference of bilirubin and glucose and found no interference at usual concentrations. The presence of abnormal hemoglobins (HbF and some Hb structural variants HbS, HbC) was not detected with the Abbott IMx assay; however, this assay showed no significant interference from the hemoglobin variants tested for heterozygous hemoglobinopathies (percentage of abnormal hemoglobin < 60%). We also determined normal values for HbAlc with this technology (164 specimens): 4.1 to 6.1%.

[The coefficient of saturation of iron in transferrin]. / A propos du coefficient de saturation en fer de la transferrine.

Theoretical iron fixation capacity of transferrin (FCT) can be calculated on its immunochemical titration: (FCT (mumol/l = transferrin (g/l) x 25). Today, its reckoning is more advisable to serum total iron binding capacity measurement. The authors studied the effects of this new proceeding upon usual values interval of transferrin saturation (i.e. serum iron/FCT ratio). The mean value and the distribution of transferrin saturation appear displaced with regard to those achieved by chemical measurement of serum total iron binding capacity. We discuss interpretation of transferrin saturation related to its methods of determination and its semiological interest.

"Sandwich" enzyme immunoassay for serum ferritin with polypropylene test tubes as the solid phase.

A "sandwich" enzyme immunoassay for ferritin in human serum is described, in which horseradish peroxidase-labeled antibody and a very sensitive chromogen, 2,2'-azino-di(3-ethyl-benzthiazolin-6-sulfonate), are used. The solid phase is polypropylene test tubes treated with glutaraldehyde; immunologic reactions are developed during constant rotation of the tubes. The assay requires 20 microL of serum per assay and takes less than 3 h. The standard curve is linear between 0 and 400 micrograms of serum ferritin per liter. Analytical recoveries of various amounts of standard added to a serum sample ranged from 92 to 98.6%. The sensitivity is 2 micrograms/L, which translates to 0.4 ng per assay tube. Within-run CV ranges from 1.6 to 5.4% and between-run from 4.2 to 7.8% according to the ferritin concentration. Results by competitive radioimmunoassay correlated well (r = 0.98). No "hook effect" was observed for concentrations up to 13 mg/L. The reference interval is 40-240 micrograms/L for men, 8-100 micrograms/L for women.

[A new 1-step immunoenzyme sandwich technic. Application to the determination of total serum IgE]. / Nouvelle technique enzymoimmunologique "sandwich" en un temps. Application au dosage des IgE totales sériques.

A sandwich enzyme immunoassay by a one step procedure for total IgE in human serum is described. This method is a modification of a two steps sandwich procedure which uses glutaraldehyde-treated tubes and antibody conjugated with peroxidase. The procedure described is characterized by immune reactions performed in one step for one hour and by a substrate solution containing orthophenylene diamine. The standard-curve is linear between 0 and 12 Ul/tube. The coefficient of within-assay ranges from 11,3 to 5% according to the concentrations. The coefficient of between-assay is 7,7% for IgE concentration of 233 kUl/l. In 107 serums, the regression equation and coefficient of correlation to another enzyme immunoassay (Phadezym IgE Prist) are: y = 0,89 X + 6,72; r = 0,98.

[Carcinoembryonic antigen. Evaluation in intestinal lavage fluids and in the tissue of colorectal tumors]. / Antigène carcinoembryonnaire. Evaluation dans les liquides de lavage intestinal et dans le tissu des tumeurs colorectales.

Carcinoembryonic antigen in pre-operative or pre-endoscopic intestinal lavage rinsing fluids was assayed in 21 healthy subjects, 15 presenting polyps and 12 presenting colorectal cancer. Comparison of the results with those of histological and immunohistological studies showed that this assay did not allow detection of intestinal cancer or evaluation of its developmental stage.

Determination of serum ferritin by a one-step immunoenzymoassay, and comparison of four liver-ferritin standards.

We describe a one step "sandwich"-type immunoenzymoassay for ferritin in human serum. The solid-phase consists of glutaraldehyde-treated polypropylene tubes coated with rabbit antibody to human ferritin. Liver ferritin is the standard. Peroxidase-conjugated antiserum to ferritin and a sensitive chromogen, o-phenylenediamine, are used. The assay requires 90 min. The standard curve is linear up to 400 micrograms of ferritin per liter of serum. Within- and between-run CVs are less than 6% for low, high, and medium concentrations and are about 13.0% at the decision level for iron deficiency. Results by a two-step "sandwich" procedure (New England Immunology Associates kit) correlated well, r = 0.98. We assessed four liver ferritin standards from different manufacturers with the described method. The mean absorbance for the 40 micrograms/L ferritin standard was 1.5 for that from Diagnostics Biochem and National Institute for Biological Standards and Controls, 1.0 for that from Dako, and 0.4 for that from Sigma. Consequently, to standardize results, all liver ferritin standards should be calibrated vs the National Institute for Biological Standards and Controls reference standard.

Serum ferritin and isoferritins are tools for diagnosis of active adult Still's disease.

OBJECTIVE: Still's disease is an acute systemic inflammatory disorder. There are no pathognomonic symptoms or specific laboratory abnormalities. Serum ferritin concentration in rheumatoid arthritis together with some plasma glycoproteins such as alpha 2-glycoprotein and C-reactive protein are part of the response to inflammation. Ferritin in plasma is glycosylated and the sialoglycosylated forms increase its microheterogeneity. Our purpose was to confirm in a large series that high values of ferritin can be found in adult Still's disease (ASD) and to see if a specific isoferritin can be isolated in this disease compared with the other systemic diseases. METHOD: Thirty-one sera were investigated from 11 men and 9 women with ASD and compared with 27 sera from 27 patients with systemic diseases. We studied the course of one case of ASD for 15 months. Serum ferritin was determined by immunoenzymology (Abbott Ferrizin). The isoferritins were investigated by isoelectric focussing and the percentage of glycosylation by affinity for concanavalin A (Con-A). RESULTS: In patients with active ASD, the ferritin levels were higher than in patients with inactive ASD or other systemic diseases: p < 0.001. The glycoforms of ferritin were basic and the proportion of ferritin bound to Con-A was lower than other ASD: p < 0.001. CONCLUSIONS: Serum ferritin levels have a diagnostic value for acute ASD. The study of sialylation and abnormalities in the glycosylation of ferritin helps to discriminate ASD from arthritis or other systemic diseases. In conclusion, the glycoform of isoferritins and the percentage of glycosylation offers an additional tool for the diagnosis of Still's disease.

[Carcinoembryonic antigen in intestinal lavage fluids and the tissue of colorectal tumors]. / Etude de l'antigène carcinoembryonnaire dans les liquides de lavage intestinal et dans le tissu des tumeurs colorectales.

Determination of carcinoembryonic antigen (CEA) was carried out in 48 intestinal washing fluids (21 from healthy subjects, 15 from patients with polyps and 12 from patients with colorectal carcinoma). The histologic study of polyps and the histologic and immunohistochemical studies of tumors were realized. In the polyps's group, the mean of ACE is significantly higher than that of the volunteers's group, but 7 subjects have a CEA level below the mean and 8 subjects have a CEA level twice this mean. Furthermore, the CEA level seemed to be independent of histologic grade of polyps. The mean of CEA levels in the subjects with colorectal carcinoma is significantly higher than that of volunteers, but 4 of 12 patients have a CEA level below the mean of healthy subjects. The CEA levels are very variable for an identical stage of tumor's differentiation and is independent of Dukes stage (B-C or D). CEA determination in intestinal washing fluids allows neither the detection nor the appreciation of intestinal carcinoma's extend.