The immunological relationship of epitopes on major tree pollen allergens.

The major allergens of birch (Bet v I), alder (Aln g I), hazel (Cor a I) and hornbeam (Car b I) were investigated by means of high-resolution two-dimensional electrophoresis combined with immunoblotting. Eleven sera derived from patients allergic to birch pollen as well as mouse monoclonal antibodies BIP 1 and BIP 4, raised against Bet v I, were used as probes. Human IgE antibodies detected 10 spots in birch (Mr 17 kDa, pI 4.9-5.9); four spots in alder (Mr 18.5 kDa, pI 4.7-5.3); four spots in hazel (Mr 17 kDa, pI 5.0-5.8); and 12 + 7 spots in hornbeam (Mr 16.5 kDa, pI 4.9-6.6 and Mr 18 kDa, pI 5.2-6.7), respectively, representing major allergens. Each patient tested reacted in a similar fashion with the spot cluster(s) of a certain allergen. BIP 1 detected the same spot clusters as patients' IgE. BIP 4 reacted with the 17-, 18.5- and 18-kDa spots of birch, alder and hornbeam, but did not react with the 17-kDa spots of hazel and the 16.5-kDa spots of hornbeam. In inhibition experiments with birch pollen extract as inhibitor, IgE binding to Bet v I, as well as to Aln g I, Cor a I and Car b I was abolished, thus suggesting that IgE binding to major tree pollen allergens is confined to shared epitopes. These findings indicate that it might be sufficient to use only Bet v I for diagnostic procedures as well as for immunotherapy in patients with tree pollen allergy.

Allergen profiles of dog hair and dander, body fluids and tissues as defined by immunoblotting.

The sera from 25 patients with clinical type I allergy against dogs were investigated by means of immunoblotting, using extracts of dog hair/dander, skin, hair, saliva, salivary gland, serum and liver. 96% of the patients' sera showed IgE antibodies reactive with 19- and 23-kilodalton (kDa) proteins in the hair/dander extract. The 23-kDa IgE-binding protein was preferentially detected in the hair extract and saliva but not in skin, salivary gland, serum and liver extracts. The 19-kDa band was strongly expressed in skin, but not in hair, serum and liver. Inhibition experiments using the 23-kDa containing extract prepared from hair and the 19-kDa containing extract prepared from skin revealed that these two proteins are likely to be immunologically independent allergens.

Diagnostic use of an analysis of urinary proteins by a practicable sodium dodecyl sulfate-electrophoresis method and rapid two-dimensional electrophoresis.

Two methods suitable for routine clinical analyses of urinary proteins are presented and compared. The first is a horizontal sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique, suitable for simultaneous analysis of 20 native urinary samples. This method uses polyacrylamide gradient gels, prepared with a laboratory-built gel casting device. The second method is a rapid two-dimensional electrophoresis procedure, combining cellulose acetate electrophoresis and sodium dodecyl sulfate-electrophoresis. The first step uses a routine system (Chemetron), the second separation step followed by staining with Coomassie Brilliant Blue R is performed on the PhastSystem. The resulting two-dimensional patterns reveal urinary proteins distributed according to the 5-zone pattern of native proteins (albumin, alpha-1, alpha-2, beta, gamma-globulin) as well as to the logarithm of their molecular weights. Examples of (routine) diagnoses with a special interest in the monitoring of kidney transplant patients are shown.

[Total cholesterol findings in Austria--an overview of epidemiologic studies]. / Gesamtcholesterinbefunde in Osterreich--ein Uberblick epidemiologischer Studien.

Hyperlipoproteinaemia is the most important public health risk factor for ischaemic heart disease. This publication gives a review of six Austrian epidemiological studies covering more than 87,000 people in all. The results are not representative for Austria, but give a picture of the epidemiology in Austria. Mean values were calculated age-standardized (age-adjusted when ten-year age groups are registered) to the Austrian population of 1985, for the age group 20 to 79 years. This age range covers 99.77% of the adult male population aged 20 or more, and 95.70% of the women. Mean total cholesterol level for the six studies is 224.2 mg/dl in men and 227.3 mg/dl in women. These results are similar to those of a representative study in the province of Vorarlberg in western Austria (220.0 mg/dl, and 227.4 mg/dl respectively). Variation range is 20.4 mg/dl in men, and 22.5 mg/dl in women; the coefficient of variation is 3.9% in both, men and women.

[The prevalence of hyperlipidemia and obesity in Vienna health examinations 1986]. / Prävalenz der Hyperlipidämie und der Obesitas bei Wiener Gesundenuntersuchungen 1986.

A random sample of 20% of people taking part in the free health check-ups provided by the city of Vienna in 1986 (707 men and 1022 women), aged 20 to 79 years (mean age men 48.1 +/- 15.8, women 49.6 +/- 15.1 respectively) were retrospectively evaluated to determine coronary heart disease risk according to the recommendations of the Austrian Cholesterol Consensus Conference. Age-standardized mean total-cholesterol is 230.0 mg/dl in men and 235.8 mg/dl in women. Age-standardized percentage of normal lipid levels (i.e. total-cholesterol below 200 mg/dl) is 29.7% in men, 25.7% in women respectively. The high-risk group (i.e. total-cholesterol more than 250 mg/dl) has an age-standardized prevalence of 28.5% in men and 35.8% in women. The age-standardized percentage of obesity according to the guidelines of the National Institutes of Health is 14.2 in men, 17.2 in women respectively. The comparison of age-standardized men-values of total-cholesterol, LDL-cholesterol, triglycerides and Quetelet-index, as well as normal lipid levels according to the Austrian recommendations and percentage of obesity, in the age groups 20 to 39, 40 to 59 and 60 to 79 years shows a statistical significant difference of each parameter between age-group 20 to 39 and age-group 40 to 59 in men, but no significant difference in any parameter between age-group 40 to 59 and 60 to 79 in men. On the contrary in women differences are significant between age-group 20 to 39 and 40 to 59, as well as between age-group 40 to 59 and 60 to 79 in each parameter.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of two solid phase chemistry systems: Reflotron and Ektachem DT 60.

Solid phase chemistry can be used for clinical analysis at the bedside, and it is even applicable to whole blood. We compared precision, accuracy, method linearity, and practicability of two solid phase chemistry analysers. Reflotron (Boehringer Mannheim, W. Germany) and Ektachem DT 60 (Eastman Kodak Co, USA). Eight analytes, glucose, cholesterol, triacylglycerols, urea, uric acid, alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyltransferase were investigated. The precision of both instruments was good. Coefficients of variation for within run and day to day precision were below 10% for all analytes. Methods were compared by analysing 88 to 105 patients sera for each investigated analyte on Reflotron, on Ektachem DT 60 and on a wet chemistry reference instrument. Linear regression analysis showed good agreement between wet chemistry and solid phase chemistry results. Coefficients of correlation (r) ranged from 0.957 to 0.999. Reflotron and Ektachem are desk top analysers. Reflotron is the smaller instrument. Currently, it offers 9 analytes and rapid single test performance. Whole blood can be used for all tests. Test strips can be stored at room temperature. Ektachem DT 60 has a modular design, and 22 analytes are available. Series of up to 100 tests per hour are possible. Whole blood can be used for the preparation of glucose and haemoglobin test slides. The slides must be deep frozen for prolonged storage. Reflotron may be suitable for the physician's office, Ektachem for small laboratories. The problem of quality control has not yet been satisfactorily solved for either instrument, as only analyser-specific control specimens can be used. Reagent costs of solid phase chemistry tests are high, especially when large test series are performed. Operation of both instruments requires well trained personnel.

Incidence of anti HIV antibodies and viral antigen in standard and control sera.

Most material used for control and calibration in a clinical laboratory is based on pool sera of human origin, guaranteed to be HBsAg-free. Since little information is available on the potential infectivity of HIV, the causative agent of acquired immunodeficiency syndrome (AIDS), 54 control and calibration sera, in routine use, were investigated for the incidence of antibodies to HIV by means of Elisa. Sixteen test specimens ( = 30%) gave positive or borderline Elisa results and were further analysed by immunoblotting, resulting in 15 samples all recognizing gp 160 and partially the p 24, p 31, p 55, p 64 and gp 120 band. Only one sample with borderline Elisa result was negative by this assay. Furthermore, all sera were examined for the presence of viral antigen by a solid phase Elisa. All samples under investigation gave negative antigen Elisa results. Bearing in mind that the sensitivity of this assay is limited to 10 micrograms/l of viral antigen, no conclusion on infectivity should be drawn. The high incidence of HIV-antibodies in the sera investigated demands that this material should be handled with special care by laboratory personnel.

Monitoring of urinary proteins by SDS electrophoresis in kidney transplant patients.

Sodium dodecyl sulphate (SDS) electrophoresis of urinary proteins was used routinely for monitoring more than 80 kidney transplant recipients as out-patients during one year. Special attention was paid to the question of whether this method can help the clinician to differentiate between a graft rejection reaction and Cyclosporin A-induced nephrotoxic damage. Two cases are presented showing the time course of proteinuric patterns together with serum creatinine, total urinary protein and, in one case, the blood level of Cyclosporin A. Changes of proteinuric patterns signalled the start of rejection and/or nephrotoxicity. A homogeneous collective of 33 kidney recipients (first transplantation) with chronic glomerulonephritis as a basic disease was specially selected. In this collective, the distribution of proteinuric patterns among patients and its dependence on immunosuppressive therapy was investigated. During the observation period of 7 months, two proteinuric patterns, "tubular" and "mixed weak", were found exclusively in Cyclosporin A-treated patients. We ascribe this finding to nephrotoxic effect of Cyclosporin A. We found the SDS electrophoresis of urinary proteins to be a useful, non-invasive method for monitoring Cyclosporin A-treated kidney transplant patients.

A simple routine method for SDS-electrophoresis of urinary proteins in kidney transplant patients.

The molecular weight analysis of urinary proteins can provide useful diagnostic information. For this purpose a routine method of sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-electrophoresis) is described. The main problem in the introduction of this method into the clinical laboratory lies in the availability of "ready-to-use" electrophoresis gels with good and reproducible quality. A device for gel production is therefore described, which is easily constructed from plastic package materials. Polyacrylamide gels are made batch-wise and in advance. They are suitable for various types and techniques of horizontal electrophoresis. Further, a method for analysis of urinary proteins is described, which permits the simultaneous analysis of 22 unconcentrated urine samples within a 3-hour-electrophoresis run. A specially optimized Coomassie blue staining method (overnight) ensures detection of proteins in the concentration range of milligrams per litre. Electrophoretic analyses were documented by photocopying the finished electrophoresis gels.

[The ISE 2020 electrolyte analyzer: results of a multicenter evaluation]. / Elektrolytanalysator ISE 2020 Ergebnisse einer multizentrischen Evaluierung.

The ISE 2020 electrolyte analyzer has been investigated within the framework of a multicentre evaluation in 5 laboratories. The users quickly learnt how to operate the instrument, and changes of personnel in the course of the trial had no influence upon the quality of the results. Regardless of the sample material (blood, serum, plasma) and the manner of introduction of the sample (manual or with the sampler), the in-series precision averaged 0.8% for sodium and 1.0% for potassium. The day-to-day precision was only slightly worse, at 1% (Na) and 2% (K). The recover, referred to the nominal values of control sera, averaged 100.3%. Investigations of linearity for potassium in the range from 0 to 12 mmol/l and for sodium in the range from 80 to 180 mmol/l confirmed a linear course of the measurements in the range of electrolyte concentrations of diagnostic interest. In extensive investigations by the testers with serum and plasma samples, the results showed good comparability with those from flame emission photometry. The mean values obtained by the two methods differed from one another only minimally in the case of sodium and potassium. The results with whole blood and with plasma prepared from it were in very close agreement. The ISE 2020 is regarded as highly suitable for routine work and emergency investigations in the central laboratory as well as for wards and operating rooms.

[Is routine bone scanning justified during the after-care for breast cancer?]. / Ist die routinemässige Skelettszintigraphie in der Nachsorge des Mammakarzinoms gerechtfertigt?

In a retrospective study of 254 women with carcinoma of the breast (mean age 55.4 years) the occurrence of bone pain was compared with results of skeletal scanning, skeletal X-ray examinations and routine biochemical findings. Typical signs of skeletal metastases were found in bone scans of 119 patients, 88 (74%) of whom had bone pain. Alkaline phosphatase was elevated in 54 (45%), LDH in 32 (27%), and gamma-GT in 69 patients (58%). There was a statistical correlation between the number of affected skeletal parts and the absolute level of alkaline phosphatase (P less than 0.001) and of LDH (P less than 0.05). Skeletal scans gave no evidence of bone metastases in 36 patients who had bone pains. In this group of patients alkaline phosphatase was elevated in 4, LDH in 1 and gamma-GT in 12 patients. Routine scanning of 254 patients revealed skeletal metastases in 12% without any clinical symptoms. Bone pain and (or) increased activity of alkaline phosphatase occurred in 91% of patients with skeletal metastases. In our view, bone scan in the postoperative control of breast cancer is justified only after onset of clinical symptoms and (or) if there is an abnormally raised alkaline phosphatase activity.

[Diagnosis of disordered iron metabolism]. / Die Diagnostik des gestörten Eisenstoffwechsels.

By use of modern methods in clinical chemistry many parameters can be selected which aid in diagnosing disturbances in iron metabolism. Each single parameter must be critically evaluated. A correct interpretation of the results is only possible in connection with clinical examinations. Determination, interpretation of relevant iron parameters and their relationship to each other by different disturbances of iron metabolism is presented.

Two dimensional electrophoresis method for additional characterization of paraproteins in serum. Clinical application of two dimensional electrophoresis, I.

High resolution two dimensional electrophoresis, one of the most potent methods for the analytical separation of proteins, is still used only in specialized laboratories. The present paper describes a two dimensional electrophoresis method, which permits characterization of abnormal serum proteins (paraproteins) and is suitable for application in a clinical laboratory. Both steps of the electrophoresis are performed by the "flat bed technique". The critical transfer from the isoelectric focusing step to the gel of the second dimension is facilitated by use of specially prepared foil stencils. The practicality of the procedure was considerably increased by using commercially available materials such as gels for isoelectric focusing and the silver stain kit. The analysis can be calibrated by measurement of the pH gradient and by using molecular weight protein standards. Moreover, two different samples can be analysed simultaneously in the same gels of the first and the second dimension. This permits a direct comparison of the sample of interest with the sample of reference. Serum samples showing an abnormal band ("M gradient") in routine methods such as agarose gel electrophoresis, were analysed by the described method. Some "M gradients" consisted of single abnormal proteins. Another sample showed a "M gradient" composed of a number of abnormal protein spots in our method. Questions concerning pathobiological and clinical evaluation of the findings justify further studies using this method.

[Significance of laboratory chemical parameters for the detection of metastases in breast cancer]. / Bedeutung laborchemischer Parameter zur Erfassung von Metastasen beim Mammakarzinom.

The diagnostic value of 7 laboratory parameters for the detection of metastases was investigated in 136 patients with verified breast carcinoma after mastectomy. The post-operative interval was 6 to 80 months (means = 27.5). 61 patients had multiple metastases as determined by physical examination, X-rays, computertomography, sonographic and scan procedures, while the other 75 patients had no evidence of metastases. Carcinoembryonic antigen (CEA), alkaline phosphatase (AP) and lactate dehydrogenase (LDH) proved to be reliable parameters for the presence of metastases; the combination of these 3 parameters had a sensitivity of 73.0% and a specificity of 94.7% in the detection of metastases. The additional determination of gamma-glutamyltranspeptidase (gamma-GT), blood sedimentation rate (BSR), C-reactive protein (CRP) and serum iron (Fe) increased the sensitivity of metastases detection to 83.8%, but the specificity decreased to 46.2%.