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Standardization of Dermatophagoides pteronyssinus extracts and tyrosine adsorbates.

Human skin test, inhibition of RAST and the radioimmunosorbent technique of Ceska have been shown to be useful assays for the standardization of D. pteronyssinus aqueous extracts. The development of these assay systems for the standardization of D. pteronyssinus tyrosine adsorbed formulations is described. Tyrosine solubilisation procedures are detailed together with the influence of these procedures on the three assays. Good correlation has been shown between the human skin test and the radioimmunosorbent technique. The inhibition of RAST appears to be more sensitive than the other techniques to mild conformational changes in the allergen, induced in this work by acid pH. This may limit its value for the standardization of D. pteronyssinus tyrosine adsorbates but it may prove useful in monitoring manufacturing processes.

Adsorption↗

Rapid non-invasive diagnosis of herpes encephalitis.

A sensitive and specific radioimmunosorbent assay for serum and cerebrospinal fluid (CSF) IgG antibodies against Herpesvirus simplex has been evaluated as an early diagnostic test for herpes encephalitis. When combined with the measurement of albumin in serum and CSF as accurate diagnostic index was obtained in twenty-four out of twenty-six patients. Out of ten patients tested before day 10 of neurological illness, five had positive diagnostic indices. Three patients with negative indices before day 10 had positive indices when re-tested later.

Antibodies, Viral↗

Serum carbonic anhydrase III in neuromuscular disorders and in healthy persons after a long-distance run.

A radioimmunosorbent technique was used for the assay of the skeletal muscle specific enzyme, carbonic anhydrase III (CA III). The usefulness of serum CA III determinations for detecting skeletal muscle damage was evaluated by comparing the serum levels of this enzyme and of myoglobin and creatine kinase in 64 patients with neuromuscular disorders and in 13 healthy volunteers before and after a long-distance run. Increased serum CA III levels were found in all patients with muscular dystrophy, chronic polymyositis and amyotrophic lateral sclerosis and in many with myasthenia gravis. In patients with polymyositis who were followed up with repeated blood sampling, the time courses of serum CA III levels, myoglobin levels and clinical symptoms were closely related. In all the runners the serum CA III level immediately after the run was increased. In the present study serum CA III and myoglobin seemed to be equally sensitive as biochemical markers of muscular damage and more sensitive than creatine kinase.

Adult↗

The interpretation of total IgE in patients with allergy.

IgE level was determined by radioimmunosorbent technique (RIST) using Phadebas IgE test in the sera of 37 patients with allergic manifestations to inhalant agents, foods, drugs and insect stings. High IgE level was found in 90% of the inhalant group and in 87.5% of the food group, whilst in the drug and in the insect sting groups the incidence was 9.1% and 0%, respectively.

Bites and Stings↗

[The effect of an isotonic solution of theophylline 300 on eosinophil cationic protein levels].

UNLABELLED: The aim of the study was to compare the influence of theophylline 300 on eosinophil cationic protein (ECP)level in serum in asthmatic patients Theophylline was given twice a day. Measurements of ECP in serum and lung function tests (PEF) were done before and at the first and fifth day after treatment Theophylline concentration were measured before and after treatment on the first injection and second injection at the first and the fifth day. ECP in serum was measured by radioimmunosorbent assay using ECP RIA kit Pharmacia. Concentration of theophylline by Emit Theophylline Assay Syva Company. The lung function test (PEF) were measured with Wright Peak flowmeter RESULT: ECP level had decreased in serum 37 + 15 micrograms/l before vs 30 + 12 micrograms/l on the first day and 17 + 9 micrograms/l on the fifth day of observation Lung function tests had improved during the observation.

Adult↗

[The effect of salmeterol on the number of hypodense eosinophils, eosinophilic cationic protein levels ECP in serum, bronchial hyperresponsiveness and lung function test in patients with bronchial asthma].

UNLABELLED: The aim of the study was to compare the influence of salmeterol xinofoate on blood hypodense eosinophil numbers ECP level in serum and sputum, and on lung function tests (VC, FEV1), bronchial hyperresponsiveness (PC20) Lung function tests (FEV1, VC), bronchial histamine challenge (PC20) and measurements of hypodense eosinophils number and ECP level in sputum and serum were done before two and six weeks after treatment with 50 micrograms salmeterol twice daily. Hypodense eosinophils were measured by Kuo et all, ECP by radioimmunosorbent assay using ECP RIA kit Pharmacia. Normal range of ECP is 2.3-15 micrograms/L Hyperreactivity by standard bronchial provocation techniques. The lung function tests (VC, FEV1, FEV1%VC) were measured with Pneumoscreen. RESULT: Eosinophil number decrease after treatment: 42.7 +/- 18.3 before vs 23.4 +/- 8.7 after. ECP level had decreased in serum 53 +/- 24 micrograms/L before and 32 +/- 12 micrograms/L after 2 weeks and 36 +/- 13 micrograms/L after 6 weeks: in sputum 2345 +/- 489 micrograms/g before and 1342 +/- 894 micrograms/g after 2 weeks 1741 micrograms/g after 6 weeks. Lung function tests had improved during the observation and hyperresponsiveness had decreased (0.08 +/- 0.1 microgram/ml before 0.24 +/- 0.2 microgram/ml after 2 week sand 0.4 +/- 0.16 after 6 weeks. CONCLUSION: ECP in serum and sputum in asthmatic patients is higher then in normal patients. Salmeterol improved lung function tests, decreased bronchial responsiveness, decreased hypodense eosinophils number and ECP concentration in serum and sputum. CLINICAL IMPLICATIONS: This date suggested that salmeterol modified bronchial responsiveness and protected the bronchial inflammation.

Adult↗

[Comparison of the bioavailability of beta-aescin after single oral administration of two different drug formulations containing an extract of horse-chestnut seeds].

The relative oral bioavailability of beta-escine (CAS 11072-93-8) from a sugar-coated tablet formulation was compared to a reference preparation available in capsule form in 18 healthy, male volunteers over a 48 h period. The study design was randomized, single-blind and cross-over. Both the test and the reference preparation contained 50 mg standardized horse chestnut seed extract; beta-escine was taken as the reference substance. By means of a newly developed, validated radioimmunosorbent assay (RIA), beta-escine in plasma was determined (blind samples) after oral intake of a single dose of each drug formulation. The confidence limits calculated for the AUC, Cmax and Tmax of the test preparation exceed the upper limit of the specified equivalence range of 80%--125%, but do never fall below the lower limit. Therefore, bioin-equivalence cannot be rejected statistically. All the bioavailability data for the test preparation--measured with the newly developed RIA--exceed the corresponding values for the reference preparation. As the rate of absorption of aesculetinic triterpene glycosides is low, the higher bioavailability of the test preparation is desirable from a therapeutical point of view. Since the reference preparation is classified as being clinically effective, the test preparation must also be estimated as being clinically effective. Adverse drug effects were not observed with either the test preparation or the reference preparation.

Adult↗

Purification of an acrosomal antigen recognized by a monoclonal antibody and antifertility effects of isoimmune serum.

A highly conserved acrosomal antigen reactive to a monoclonal antibody (HS-63), generated against human sperm, was purified to homogeneity with a combination of conventional procedures and immunoaffinity chromatography using a soluble extract of mouse and rabbit testes. The molecular weight of the purified antigen was 42-50 kD when analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis. The high specificity of the purified antigen to monoclonal antibody HS-63 was shown by indirect immunofluorescent inhibition assay, enzyme-linked immunosorbent assay, Western blot analysis and radioimmunosorbent assay. The purified antigen was used for isoimmunization of mice and rabbits. Following successive immunizations, antisera of high titres were raised and reacted specifically with antigen on the sperm acrosome and in testes of several mammalian species, but not with somatic tissues. These isoimmune sera exhibited strong inhibition on mouse in-vitro fertilization and human sperm penetration of zona-free hamster eggs. The results of this study suggest that the sperm-specific acrosomal antigen reacting with HS-63 could be a good candidate for the development of immunocontraceptive vaccines in humans and in other animals.

Acrosome↗

Beta-2-microglobulin as an index of renal function after cardiopulmonary bypass surgery in children.

In order to evaluate the clinical usefulness of serum and urinary beta-2-microglobulin (beta 2-m) determination as a marker of renal damage following cardiopulmonary bypass surgery (CPB), 37 children, with an age range of 6 months to 13 years undergoing CPB under hypothermia and cardioplegia for congenital malformations, were studied. Renal function was monitored at baseline and on the first and third day after CPB by traditional tests such as creatinine (Cr), endogenous creatinine clearance (Ccr), and fractional sodium excretion (FeNa), as well as by serum and urinary beta 2-m measured by radioimmunosorbent assay. Data were analyzed for the group as a whole and after stratification for the presence of acute renal failure (ARF) defined by an increase in Cr greater than 0.5 mg/dl and/or FeNa greater than 2% and of renal hypoperfusion which was considered in case of hypotension resistant to volume repletion and/or dopamine infusion and of oliguria (less than 1.0 ml/kg/h). The incidence of ARF was 18.9%, mortality 21.7% and among those who developed ARF, 71.4%. Although Cr and FeNa increased significantly on the first postoperative day, values returned to baseline thereafter, probably because of the high mortality rate observed among those who developed ARF and were therefore lost to follow-up. In contrast, Ccr was significantly decreased during the whole study. Even though serum beta 2-m displayed a similar profile to Cr and FeNa returning to baseline by the third day, urinary and fractional excretion of beta 2-m were significantly increased at any time of the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Kidney Injury↗

Immune recognition of human major histocompatibility antigens: localization by a comprehensive synthetic strategy of the continuous antigenic sites in the first domain of HLA-DR2 beta chain.

A comprehensive synthetic approach, previously developed in this laboratory, has been applied to screen the entire first domain (residues 1-95) of the HLA-DR2 beta chain for the full profile of continuous regions that are recognized by human allo-antisera and anti-DR or anti-beta chain antisera raised in other species. Nine consecutive peptides, that were 15 residues each and overlapped by 5 residues, covering the first domain of the DR2 beta chain were synthesized, purified and characterized. The antibody-binding activities of the peptides were determined by radioimmunosorbent titrations. This established the full profile of peptides having specific antibody-binding activity with these various antisera. Three continuous antigenic sites were localized in this domain by all of antisera tested. Peptide 81-95 appeared to be the most immunodominant (i.e. bound the highest amounts of antibodies) in most antisera tested. Although their immunodominance varied with the antisera and boundary shifts were present, the submolecular regions recognized on the first domain of DR2 beta appeared to be similar with antisera against the DR2 (alpha beta I) complex or against the isolated beta chain. Furthermore, recognition was independent of the host species from which the antisera were obtained.

Animals↗

Carbonic anhydrase in the human fetal lung.

Lung tissue from human fetuses, with gestational ages ranging between 14-26 wk, was studied by histochemical and biochemical methods. The findings were similar in all tissues tested, without apparent correlation to gestational age. Staining that indicated carbonic anhydrase activity was found in the capillary endothelium and in the epithelium of some segments of the peripheral airways. The ciliated epithelium of the central airways was unstained. The distribution of the enzyme in the human fetal lung differed clearly from that in the adult human lung, where little or no enzyme has been found in the airway epithelium. The mean carbonic anhydrase activity in whole homogenates of fetal lung tissue was 24 enzyme units per g wet weight of tissue. Ninety % of this activity was recovered in the supernatant fraction. Assay of this fraction by a radioimmunosorbent technique showed the presence of the carbonic anhydrase isoenzyme HCA-C corresponding to 380 ng enzyme per mg tissue protein. Small amounts of HCA-B were also found but are thought to be attributable to contaminating erythrocytes; thus, the data suggest that both the capillary endothelium and the lung epithelium contain HCA-C, an isoenzyme of carbonic anhydrase known to be involved in electrolyte transport in many tissues.

Bronchi↗

Variation in surface platelet glycoprotein Ib expression in Bernard-Soulier syndrome.

Seven patients with Bernard-Soulier syndrome (BSS) and 15 presumed heterozygotes of BSS from four families are presented. Evaluation of their platelet membranes was performed by an enzyme-linked radioimmunosorbent assay (ELISA) technique including monoclonal antibodies specific for glycoprotein (GP) Ib and GPIIb/IIIa. Analyses of platelets from the patients revealed 6-33% of the normal GPIb concentration; siblings showed nearly equal amounts of this component. One of the relatives had 44% of the normal level, while 85-98% (mean 92%) of the normal GPIb content was observed in the remaining relatives. Normal or slightly elevated levels of GPIIb/IIIa were detected in the patients as well as in the relatives. All relatives had normal platelet count, size distribution, bleeding time, and ristocetin-induced platelet aggregation. The patients and their relatives may account for a biological variation in GPIb expression or may represent a variant type of BSS.

Adolescent↗

Serum IgE in atopic dermatitis: relationship to severity of cutaneous involvement and course of disease as well as coexistence of atopic respiratory diseases.

Serum IgE concentrations were determined according to the radioimmunosorbent technique (RIST, Phadebas) on 116 adult patients with atopic dermatitis of varying severity and activity. Geometric mean IgE levels of patients with atopic dermatitis were significantly higher compared with the mean level of ninety-three non-atopic adult subjects without parasitic infestation. Severity of the atopic dermatitis was highly correlated to the levels of serum IgE. Severe chronic cases with ever-recurrent exacerbations show the most extreme values. In the moderate forms of atopic dermatitis, coexistent bronchial asthma causes a greater increase in the IgE values. Among the mild or abortive forms higher IgE levels were found in cases with allergic rhinitis than in the cases with 'pure' atopic dermatitis. Other findings in connection with IgE in atopic dermatitis are summarized. The pathogenetic significance of IgE in the cutaneous changes is briefly discussed.

Adolescent↗