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Resume A l'aide de resultats experimentaux obtenus en allergologie, on definit quelle strategie experimentale doit etre utilisee, lors-qu'on etudie le lien pouvant exister entre une symptomatologie et un parametre biologique mesurable (test biologique), ou lorsqu'on desire evaluer si ce test biologique peut etre pratique dans un but diagnostique. Dans le premier cas, les etudes de comparaisons de moyennes correspondant a des groupes d'individus differents vont permettre de repondre a des questions sur la relation entre un type de pathologie et la valeur experimentale des parametres mesures, analyses qui peuvent etre ensuite approfondies par des etudes de correlation. Dans le second cas, sont definis les criteres retenus comme indispensables pour valider un test biologique vis-a-vis du diagnostic clinique, notamment la sensibilite et la specificite. A partir de ces deux criteres, elabores grâce a l'etude de populations bien definies (malades et temoins), on peut calculer les valeurs predictives positive et negative en tenant compte de la prevalence de la maladie. Alors, le resultat d'un test biologique pour un individu appartenant a un echantillonnage reel de consultants peut etre interprete comme normal ou pathologique en fonction de sa situation vis-a-vis de ces normes de reference.

Background Allergy to banana often occurs in patients sensitized to latex. The spectrum of IgE-mediated responses to latex allergens is more and more documented but banana allergens and epitopes shared by these two allergens have not yet been characterized, even though the existence of cross-reacting IgE antibodies has been demonstrated. Objective The purpose of this study was to assess the relationship between banana hypersensitivity and latex allergy in a population of 19 latex-allergic patients and to identify allergenic components in banana and the common structures shared with latex. Methods The in vivo study was conducted in our outpatient department in patients with well-documented latex allergy. Clinical histories were evaluated and skin prick tests were performed with banana and latex extracts. IgE responses from 19 patients were investigated by means of CAP RAST assay and SDS PAGE immunoblotting. Epitopes shared by banana and latex were investigated by means of immunoblotting experiments. Results Eight of 16 patients (50%) reported symptoms after eating bananas and banana skin prick tests were positive in 5 of 14 patients (36%). Banana RAST results were positive in 12 of the 19 patients (63%). In immunoblot experiments, 17 of the 19 patients (89%) exhibited specific banana IgE antibodies and 16 allergenic components were identified with molecular weights ranging from 17 to 128 kD. Two were considered as major allergens: 33 kD was detectable in 15 of 19 sera (88%) and 37 kD in 13 of 19 sera (76%). Inhibition studies by preincubation of two individual sera with banana or latex extract demonstrated the complete disappearance of IgE binding on banana blotted allergens. Conclusion This study confirms the "latex-fruit syndrome" already described by Blanco et al. Two major allergens were revealed in banana at 33 and 37 kD and the presence of more than ten common components with latex was observed.

Resume L'indication et l'interpretation du dosage serique des IgE et des IgG specifiques ainsi que du test d'histaminoliberation in vitro sont discutees comme elements de diagnostic de l'allergie aux venins d'hymenopteres. La place de ces tests dans le bilan conduisant a la mise en œuvre et au suivi d'une desensibilisation est evaluee.

Urokinase type plasminogen activator (uPA) and its inhibitors, plasminogen activator inhibitor type I (PAI-1) and type II (PAI-2), are supposed to be involved in the expression of the invasive and metastatic phenotype of cancer cells. However, clinical investigations on the prognostic significance of their levels in tumor tissue are difficult to realize because of the absence of a convenient method of measurement of these parameters. The aim of the present investigation was to set up a method allowing the measurement of these enzymes and of sex steroid receptor status in appropriate subcellular fraction(s) in conditions easily reproducible in routine. We found that a tissue homogenate prepared according to the method recommended [5] for current measurement of sex steroid receptors is appropriate for further distinct preparations. One aliquot is used for cytosol preparation; another can be treated by 2% Triton X-100 (vol/vol) and provide an extract containing the totality of uPA and PAI-1. The advantage of this procedure is that appropriate subcellular fractions can be derived from a unique homogenization step. Total uPA and PAI-1 are measured in a Triton extract with good performance as compared to previous investigations [4]. PAI-2 is measured in the same cytosol fraction used for sex steroid receptors and other parameters. Because of its simplicity and its high reliability, this method could be a useful tool in the investigation of uPA family proteases and analysis of their prognostic significance in early breast tumors.

Association between allergy to Ficus benjamina and Hevea brasiliensis, two botanically unrelated plants, was suspected in consequence of two clinical observations. Symptoms were rhinitis and asthma. This study was undertaken to assess the in vivo and in vitro cross-reactivity between Ficus benjamina and Hevea brasiliensis allergens.The two patients were asked about use and contact with latex devices and relationship between symptoms and Ficus benjamina exposure. Skin prick tests were performed with Ficus benjamina, Hevea brasiliensis extracts and common allergens. Double-blind nasal and bronchial challenge tests were done using the rinse fluid from a brand of latex gloves. Total and specific IgE antibodies to Ficus benjamina and Hevea brasiliensis were determined. In vitro cross-reactivity was investigated by means of CAP RAST and immunodot inhibition experiments.We observed that for the first patient the primary phenomenon is probably allergy to latex followed by allergy to Ficus benjamina. For the second patient, allergy to Ficus benjamina was diagnosed (improvement related to the avoidance of exposure to Ficus benjamina allergens) and positivity to latex skin prick tests may be due to the cross-reacting allergens. In vitro assays showed specific IgE antibodies to both allergens and cross-reactivity was confirmed in the two cases by reciprocal inhibition of the two extracts.The increasing risk of sensitization to widely used latex devices and extensive exposure to Ficus species in households and offices indicates increased allergenic risk from this newly recognized cross-reactivity.

Different methods are available for cobalamin determination in serum. Microbiological and radio ligand binding assays are the most commonly used. Kits involving non-isotopic competitive-binding assay have been recently commercialized. In the present work, cobalamins were determined in 146 patient sera, using four methods: a microbiological method, two no boil radio ligand binding assay kits (Magic B12 FOL (NB) from Ciba-Corning and SimulTRAC SNB No Boil from Becton Dickinson) and a non-isotopic kit with acridinium ester labelled cobalamin (Magic Lite from Ciba-Corning). Median (range) cobalamin concentrations in pmol l-1 were 317 (15-1291) using the microbiological method, 355 (25-3469) using the Magic Lite kit, 355 (35-2312) using the Magic B12 FOL (NB) kit and 380 (37-2021) using the SimulTRAC SNB No Boil kit. The ANOVA test indicated that differences between methods were statistically significant (p < 0.01). Competitive-binding methods gave higher results than the microbiological method. Although correlation coefficients were not excellent (0.88 < r < 0.96), the results obtained with the different methods were generally similar and confirmed that competitive methods are useful for detecting low serum concentration of vitamin B12.

Folates were determined in 148 patient sera, using four different methods: a microbiological assay (Reference Method), two radioassays (Magic B12 FOL [NB] and SimulTRAC SNB no boil kits) and a non isotopic competition method (Magic Lite kit). Folate mean varied according to the methods 10.0 nmol.l-1 (reference method), 12.2 nmol.l-1 (Magic Lite), 8.7 nmol.l-1 (Magic B12 FOL [NB]) and 10.8 nmol.l-1 (SimulTRAC SNB NO boil). Poor correlations were also noted (0.83r0.95, figures 1 and 2). Differences between methods were explained according to the literature.

Results of immunization against hepatitis B among Pasteur Institute staff members are reported. Prior to immunization, 439 subjects were tested for hepatitis B virus (HBV) markers, including HBs antigen, anti-HBs antibody, and anti-HBc antibody (Ausria, Ausab, Corab assays; Abbott). Forty-seven subjects tested positive for anti-HBs antibody. 317 subjects negative for all the HBs markers studied were given three intramuscular doses of Hevac B (Pasteur vaccins) at one-month intervals. Anti-HBs antibodies were assayed after the third injection with the following results: mean titer, 1,454 mIU/ml, standard deviation, 5,349 mIU/ml, and range, 4 to 41,100 mIU/ml. Anti-HBs titers above 10 mIU/ml were found in 879.4% of subjects. Non-responders and weak responders (anti-HBs titer under 10 mIU/ml) were given a fourth dose of vaccine. Ultimately, after the last (third of fourth) injection 97.6% of subjects had protective antibody titers. No case of HBV infection was seen during the seven-year follow-up period.

The aim of this study was to compare serum N-terminal peptide of type III procollagen to aminotransferases and gamma-globulins as a marker for histological activity in patients with chronic hepatitis and to assess the role of type I collagen, a new serum marker, as a marker of fibrosis in these patients. Sixty patients with biopsy-proven chronic hepatitis were included in this study. Liver disease was virus B-related in 29, autoimmune in five, drug-induced in five, and of unknown etiology in 21. Each biopsy was independently assessed by two liver pathologists. Two histological scores, a score of activity and a score of fibrosis, were established. Serum N-terminal peptide of type III procollagen and type I collagen were assayed by liquid phase RIA. Significant correlations were noted between serum N-terminal peptide of type III procollagen and scores of activity (r = 0.70, p less than 10(-4)) and fibrosis (r = 0.45, p = 0.0005), and between serum type I collagen and scores of activity (r = 0.46, p = 0.0004) and fibrosis (r = 0.67, p less than 10(-4)). When the correlation between scores of activity and fibrosis (r = 0.52, p = 10(-4)) was considered by partial correlation, serum N-terminal peptide of type III procollagen was correlated with the score of activity (r = 0.63, p less than 10(-3)) but not with the score of fibrosis, and serum type I collagen was correlated with the score of fibrosis (r = 0.58, p less than 10(-3)), but not with the score of activity.(ABSTRACT TRUNCATED AT 250 WORDS)

The serum concentration of aminoterminal propeptide of type III procollagen was measured in 44 Egyptian healthy controls and 29 patients with hepatosplenomegaly originating from endemic areas for schistosomiasis in Egypt. Patients were classified into two main groups according to the histopathological pattern of the liver biopsy: patients with active schistosomal liver fibrosis and patients with inactive schistosomal liver fibrosis. Serum aminoterminal propeptide of type III procollagen levels were elevated in most of patients with active fibrosis but not in those with inactive schistosomiasis. From the present work, it is suggested that aminoterminal propeptide of type III procollagen can be used as a marker for active fibrogenesis in patients with schistosomal liver fibrosis.

The analytical performances of the Abbott IMx for the detection of several markers of hepatitis A and B (HBs Ag, anti-HBc, IgM anti-HBc, IgM anti-HAV) were evaluated. The intra-assay and the inter-assay coefficients of variation were below 5 and 10% respectively. We obtained results comparable with our routine methods, except for sera containing rheumatoid factor which gave false positive reactions with the IMx HBs Ag.

This radioimmunoassay for type I collagen mainly detects degradation products of the molecule in human serum samples. Type I collagen antigenicity in serum can be separated into two peaks by gel-filtration chromatography. The larger form represents collagen molecules (as shown by immunoblotting experiments), and (or) type I collagen with aminoterminal propeptide or intact procollagen molecules. The smaller form, the exact nature of which is not known, is quantitatively the principal antigenic form and is derived from degradation of type I collagen. The concentration of type I collagen in serum is increased mainly in cirrhotic patients, with or without active liver disease, but also somewhat in alcoholic patients without cirrhosis.

Ruthenium (III) tris acetylacetonate is an efficient catalyst for the isomerization of O-deuterated alkylvinyl carbinols allowing thus a facile route to the title compounds.

Summary To detect a foreign collagen in a tissue on a substantially idemtied protein, a radio-tracer technique appeared ideal. The method is only valuable if the introduction of the tram element into the micro molecule does not modify its properties. The first collagen used was tagged by the introduction of C1* into the peptide chain from marked proline by biosynthesis. A 0.3% concentration in sorbic acid solution was applied to the back of the animal and the penetration followed by measuring the activity of tissue sections. No radioactivity was found in the interior of the deinnir after periods of 15, 30,90 and 120 min after application. The method of biosynthetic marking was abandoned as the activity of the protein was considered to be insufficient. The technique of the incorporation of 1(125) in the tyrosine was used to obtain a greater specific activity without modifying the active principle. This chemical treatment resulted from the oxidation of tagged iodide in the presence of collagen. The protein was purified by chromatography and successive dialyses and diluted in unlabelled collagen for subsequent manipulation. Skin applications were made in sorbic acid solution at 0.3% and at 10% in a cosmetic cream. Skin penetration forthe same periods as previously were followed by the same sectionning technique. The penetration reached a maximum at the end of 15 min and was greater for the cream than for the solution. Metabolic studies showed that 15 days after application the residual radioactivity was present mainly in the lymphatic glands and in the skin. Treated dcin showed more activity than untreated areas. At the end of 10 days the excretion of radioactivity had dropped to very low levels. Chromatography on molecular sieve of the collagen extracted from the deeper layers on the skin indicated that the largest part of the radio-activity originated in peptide chains.

Evidence is presented for the reversible formation of metal-alkyne complex in the pyrolysis of eta/sup 5/-cyclopentadienyl-eta/sup 4/-cyclobutadiene-cobalt complexes. (DLC)

Resume Phadiatop™ est un nouveau test biologique permettant le depistage de l'allergie respiratoire. Ce test, base sur le principe du RAST, utilise un disque de papier sur lequel est fixe un melange equilibre de pneumallergenes selectionnes. 105 enfants (4 mois — 15 ans) d'une consultation d'allergologie ont ete etudies. Il existe une bonne correlation entre ce nouveau test et le taux des IgE totales (70,5 %), des IgE specifiques (96,2 %) et les tests cutanes (95,2 %). La correlation entre le diagnostic clinique et les resultats du Phadiatop™ est hautement significative : efficacite 98,1 p. cent, sensibilite 96,1 p. cent et specificite 100 p. cent.

A solid-phase radioimmunoassay using 125I-protein A is described for the detection of antibodies to collagens of different types. The optimal conditions for the adsorption of collagen onto polystyrene microplates, then the incubations with the antiserum and finally with the 125I-protein A have been evaluated. The technique was applied successfully to antisera raised in rabbit, goat, guinea pig and mouse against human type I, II, III, IV, V and bovine type I, II, 1 alpha 2 alpha 3 alpha, X1-X7 collagens.

The authors discuss about the functions and the evolution of total specific IgG antibodies and specific antibodies of the IgG1 and IgG4 isotype during desensitization treatment.

The anterior cruciate ligament was severed in the right knee of 36 rabbits which were then killed 12 weeks after surgery. Changes in water content and fixed charge density were correlated with the surface area of induced cartilage fibrillation. Histological changes including cartilage hypertrophy, reduced cell density, matrix alterations, cystic lesions, and enlarged perichondrocytic lacunae were found to precede cartilage fibrillation. The observed histological changes are in agreement with an enzymatic degradation of articular cartilage in osteoarthritis.

The authors have developed a solid phase radioimmunoassay for the detection of antibodies to collagens. They give the technical data of this assay and some results mainly in rheumatic diseases.

In the present work a study has been made of the prevalence of allergy to two storage mites, Tyrophagus putrescentiae and Lepidoglyphus destructor in a population of 105 young adults in the east of France. Combination of skin tests and specific IgE measurements shows that 43.10% of the subjects were sensitive to Tyrophagus and 44.95% to Lepidoglyphus. Interpretation of skin tests remains uncertain, particularly concerning the source of the allergen used and it is most often also necessary to measure the specific serum IgE. As in many studies, most sensitization was found amongst farmers. In contrast, no difference was seen between rural and city dwellers, subjects who were in contact with small animals and those in contact with food for livestock if they were not farmers, not between declared asthmatics who lived in a humid place and those who lived in an environment that was drier. Certainly these mites appeared in the present study to be much more ubiquitous than supposed previously and the fact that they seemed to be carriers of allergens and antigens that were different from those of Dermatophagoides raises the question as to whether the need for hyposensitization treatment for them has been under-estimated.

A small percentage of patients treated with bovine collagen implants have adverse reactions involving both the cellular and humoral types of immune response. We report a clinical follow-up of 705 subjects treated with a new bovine collagen implant, Atelocollagen (Koken Co. Ltd., Tokyo, Japan). Sensitization to the implant was evaluated in all subjects by skin testing, and humoral response was monitored in 166 subjects by measuring the level of circulating antibodies directed against bovine collagen. Twenty-seven patients (3.8%) exhibited a positive response to a skin test, and of the remaining 656 patients, an adverse reaction to the implant developed in 2.3%. We found a strong correlation between the presence of antibodies to collagen and a positive response to skin testing (92%) or an adverse reaction (100%). In the case of a borderline clinical response to bovine collagen implantation, anticollagen serologic tests appeared to be a useful tool for the identification of clinically reactive patients.

We report a radio-immunoprecipitation method for the measurement of circulating insulin antibodies using porcine, bovine or human radiolabeled insulin and precipitation of the bound fraction by a combined polyethylene glycol--second antibody method. The coefficient of variation for intra- and inter-assay is below 8,5 p. cent and 17 p. cent respectively. 100 insulin treated patients sera were tested in the three systems: the results showed a great scattering; for the same serum, the values obtained with porcine and human insulins are below that with bovine insulin. The data correlated statistically significantly with those found with a commercial kit (r = 0,97, n = 170, t = -4,91, p greater than 0,001).

The present paper describes the optimisation of a liquid phase [125I] C1q binding assay for the detection of circulating immune complexes. Blood must be collected without any anticoagulant. The serum can be stored or transported as usual without damage. A 90 min incubation is needed to reach the equilibrium of the reaction; polyethylene glycol (2,5% final concentration) is added to precipitate the [125I] C1q-CIC complex. By its simplicity and reproducibility, this test can be currently employed, especially in the case of treatment by plasmapheresis.

Rapid, specific and reproducible radioimmunoprecipitation assay (RIP) for measuring IgG antibodies against Yellow Jacket Venom (YJV) has been developed. The YJV-I125 is added to serum sample and the immune complexes are precipitated by a second antibody with polyethylene glycol. IgG antibodies were measured by RIP in 24 patients receiving Yellow Jacket Venom immunotherapy. RIP assays were compared for 159 sera to a solid phase procedure (RAST IgG).

Fibronectin (by laser nephelometry) and collagen IV blood levels (by radioimmunoassay) were studied in 183 diabetics and compared with 101 non diabetic patients. Diabetics have more collagen IV and less fibronectin than non diabetics. Divergence has increased since 20 years old; fibronectin blood levels is always low in diabetics, even in the young. Collagen IV is higher in diabetics with angiopathy, and specially if insulin dependent. Diabetics which have a good control have normals levels. In the other hand when HBA1C greater than or equal to 9%, collagen IV blood level increases quickly and fibronectin level decreases. The importance of the antidiabetic treatment is underlined.

The authors present an experimental study of the rabbit femoral prosthesis compatibility : macroscopical aspect, proteoglycan content and metabolism of the acetabular cartilage.