Human serum alpha 2HS-glycoprotein modulates in vitro bone resorption.

We previously isolated a family of bone-resorbing proteins from human cancer ascites fluid and established that the three purified bone-resorbing proteins were chemically and immunochemically related to each other and to alpha-2HS glycoprotein (alpha 2HS). After this finding we purified the normal human serum counterpart of these ascites proteins and studied its effects on bone resorption. The bone-resorbing properties of normal human serum alpha 2HS were examined in vitro over a wide dose range. This normal human serum glycoprotein had a biphasic effect on 45Ca2+ release from bone. More specifically, this protein stimulated bone resorption at the lower concentrations tested, with a maximum effect [treated over control ratio of 2.5 +/- 0.30 (+/- SE); P less than 0.01] at 40 micrograms/mL. In contrast, at doses above 40 micrograms/mL, a sharp decline in calcium mobilization occurred, with a return to baseline occurring above 80 micrograms/mL. These results suggest that serum alpha 2HS may participate in the regulation of bone metabolism in vivo.

One-step sandwich enzyme immunoassay for insulin using monoclonal antibodies.

An enzyme-linked immunosorbent assay for the measurement of insulin in human serum has been developed. The test is based on the sandwich technique with two monoclonal antibodies directed against two different epitopes of insulin using coated plastic tubes as the solid phase and horse radish peroxidase as the label. The immunoreactions are completed in one step within 2 h. The horse radish peroxidase activity bound to the tube wall is measured photometrically after an additional 1-h incubation with the substrate. The standards used cover the range from 0 to 260 mU insulin/L. Employing the Enzymun-Test System ES 22 modular batch analyzer, the detection limit was found to be 3.7 mU insulin/L. Coefficients of variation (CV's) between 1.4-7.8% for intraassay precision and 5.6-10% for interassay precision were obtained over the concentration range of 17-107 mU Insulin/L. The correlation between the procedure described here (y) and a commercially available double antibody radioimmunoassay (x) is expressed by the following equation: y = 1.07x + 1.14 mU insulin/L.

Behavior of beta 2-microglobulin in patients with chronic renal failure undergoing hemodialysis, hemodiafiltration and continuous ambulatory peritoneal dialysis (CAPD).

beta 2-microglobulin (beta 2-m) is the major component of a new form of amyloid deposit found in carpal tunnel syndrome and dialysis arthropathy of long-term hemodialysis patients. In 52 patients on maintenance hemodialysis, serum beta 2-m concentration was elevated to 37.9 +/- 1.4 (normal 1.2 +/- 0.6) mg/l. It was correlated with the time on hemodialysis (r = 0.43, p less than 0.01) and was inversely correlated with residual renal function (r = 0.87, p less than 0.001). In 20 patients on CAPD, beta 2-m likewise was increased to 31.6 +/- 2.3 mg/l; daily elimination by dialysate was only 34 mg (normal 150 mg). Hemodialysis with a cuprophane membrane caused a rise in serum beta 2-m, whereas hemodiafiltration with a polysulfone membrane performed in 5 patients over 2 1/2 months was accompanied by a decrease in serum beta 2-m from 39.5 +/- 0.7 to 29.7 +/- 1.0 mg/l predialysis (19.1 +/- 1.1 postdialysis). On the other hand, beta 2-m elimination reached only approximately 100 mg per day in spite of markedly elevated serum levels. It is concluded that serum beta 2-m is massively elevated in long-term hemodialysis and CAPD patients; contrary to routine hemodialysis with cuprophane membranes, newer more permeable membranes will permit some elimination of beta 2-m. However, based on quantitative considerations it seems difficult to obtain beta 2-m concentrations in the high normal or moderately elevated range with present day techniques.

[The hematological laboratory of clinical practice]. / Das hämatologische Praxislaboratorium.

The spectrum of analyses of a hematological laboratory comprises the counts of erythrocytes, leukocytes and platelets, the determinations of the hemoglobin concentration and of the hematocrit, the calculation of the red cell indices and the microscopic examination of the blood film. The determination of white blood cells, platelets, hematocrit and MCV as well as staining of the blood film should be performed within two hours after blood sampling. The hemoglobin concentration, red-cell count and MCH value remain constant for five days, provided that the blood samples is stored at 4 degrees C. The recognition of morphologically 'minor' pathological findings is important in view of the possible clinical significance that they might have. In the case of thrombotic-thrombocytopenic purpura, for instance, it is very important that the fragmentocytes are recognized and recorded. The grading of pathological findings by use of +, ++, is based on strict limits of ranges. Quality control and quality assurance are integral parts of the daily laboratory performance; moreover, the importance of external quality control in morphological hematology is stressed as being a challenge to the hematological laboratory technologist.

[Platelet satellitosis]. / Dostickový satelitizmus.

Platelet satellitism (PS) is a rare phenomenon described sporadically in the literature. It involves adhesion of thrombocytes sensitized by antibodies to leucocytes and is observed in vitro in EDTA anticoagulated blood. The authors' contribution to the problem is the observation of special thrombocyte aggregates surrounding neutrophils resembling comet tails, as well as the fact that the authors observed the formation of aggregates surrounding also lymphocytes and eosinophil cells.

[Hemolysis-inducing substances in gastric secretion, incidence-rate of lysolecithin]. / Hämolysierende Substanzen im Magensekret, Häufigkeit des Lysolecithin-Nachweises.

Samples of gastric contents from 152 patients with pyloric reflex were taken during gastroscopy after an overnight fast and examined for hemolytic activity. Hemolysis was induced by 97 specimens (64%). The hemolysis test was positive in 45% of patients with a histologically normal gastric mucosa, in 76% of patients suffering from chronic atrophic gastritis with intestinal metaplasia, in 83% with gastric erosions and in 67% with gastric ulcer. No lysolecithin was found in 9 of the 97 positive specimens. The other aspirates contained widely differing values up to 320 mg/100 ml. The average quantities of lysolecithin in gastric contents varied in the different patient groups from 20 to 60 mg/100 ml. These values are much higher than the mean value of 0.9 mg/100 ml quantified in patients without pyloric reflex during an earlier investigation. It is now widely accepted that pyloric reflex promotes gastritis. Furthermore, it has been shown on several occasions that bile constituents exert a damaging effect on the gastric mucosa barrier. The same is true of lysolecithin, which promotes (for example) acute cholecystitis under experimental conditions. These findings, together with the results of our investigation, seem to afford evidence that lysolecithins may exert a pathogenic influence in the development of different gastric lesions.

Highly elevated content of free myo-inositol in the cerebrospinal fluid of neonates.

Free myo-inositol in cerebrospinal fluid was determined in 60 babies aged from newborn to 12 months. In the neonates, a high inositol concentration was found (18-81 mg/100 ml). With increasing age, the values gradually decreased. The finding is discussed in relation to the maturation processes in central nervous tissue.

[Dry chemistry in the clinical laboratory: how reliable is it?]. / Trockenchemie im Praxislaboratorium: wie zuverlässig?

The performance of dry chemistry analysis systems was evaluated using the results obtained from proficiency testing 1991 of practitioner's laboratories. 59% of the participants use dry chemistry (Ektachem DT 60 Kodak, Reflotron Boehringer, Cobas Ready Roche). The evaluation reveals the following: The declared values of the control sera for the dry chemistry systems agree well with those of the conventional wet chemistry, excepting enzyme assays. The precision of the dry chemistry parameters is substantially higher than that of the wet chemistry. The overall performance of the dry chemistry analysis systems was found to be superior as compared to the wet chemistry performance. A prerequisite for dry chemistry proficiency testing are control sera of human origin.

[Non-enzymatic marking of gene probes using digoxigenin for the diagnosis of sickle cell mutation and beta thalassemia]. / Nicht-enzymatische Markierung von Gen-Sonden mit Digoxigenin für die Diagnose der Sichelzellmutation und der beta-Thalassämie.

A method is described in which allele-specific oligonucleotides (ASO) are labeled non-enzymatically with digoxigenin (DIG). The ASOs were synthesized in a DNA synthesizer. A primary C6-alkylamine was incorporated at the 5'end of the ASO. The derivative was coupled to digoxigenin-3-0-methylcarbonyl-epsilon-aminocaproic acid-N-hydroxy-succinimide ester. The DIG-ASO probes were isolated from the crude preparation and hybridized with amplified genomic DNA. The annealed probes were detected with an ELISA test. The DIG-ASO probes were successfully used for the identification of sickle cell disease genotypes and the most common beta-thalassemia mutations.

[Isolation, identification and quantitative determination of lysolecithin in the human gastric juice]. / Isolierung, Identifizierung und quantitative Bestimmung von Lysolecithin in menschlichem Magensaft

Lysolecithin was isolated and identified from the gastric contents of 5 patients with gastric ulcer and erosive and atrophic gastritis. The methods used involved methanolic extration, column chromatography on silica gel and sephadex gel filtration, followed by preparative thin layer chromatography. Identification of lysolecithin was verified in one sample by thin layer chromatography and elementary analysis. A method for quantitation of lysolecithin in human gastric fluid for routine clinical analysis is described, based on the procedure used for identification of this phospholipid. Further, a hemolysis test is proposed for screening gastric contents for lysolecithin concentrations above approximately 10 mg/100 ml. With the quantitative method, an average concentration of 66.1 mg of lysolecithin in 100 ml gastric content was found in the group of 5 patients. The screening test was positive in all 5 samples, but was negative with native gastric juice in 25 persons serving as controls. When the gastric fluid of the controls was concentrated, however, 9 samples gave a positive hemolytic reaction. From these 9 specimens, the lysolecithin concentration was found to average 0.9 mg/100 ml. These findings afford evidence that lysolecithin may be involved as a factor in the pathogenesis of gastric ulceration.

[Elimination of fluoride in urine during fluoridation of salt and drinking water]. / Fluoridausscheidung im Urin unter Salz- und Trinkwasserfluoridierung.

From 552 ambulatory gynecological patients in different parts of Switzerland the urinary excretion of fluorine was measured. The patients were classified into 4 groups according to origin: (1) 84 women were from the Canton Basel-Stadt. In this city water has been fluoridated since 1962; (2) 139 women from Canton Glarus. In this area a pilot study was under way using table and baker's salt, to both of which 250 mg F/kg had been added; (3) 128 women from Canton Aargau who were using a low dose fluoridated salt (90 mg F/kg); (4) 201 patients from Cantons Aargau and Tessin respectively who were consuming neither fluoridated water or salt acted as controls. Quantitation of ionized fluorine in urine was performed by means of the fluoride ion sensitive electrode in afternoon urine samples, thus eliminating the influence of sex difference and diurnal rhythm in fluorine excretion. The molar urinary fluorine concentration was related to the corresponding urinary creatinine concentration and expressed as mumol F per mmol creatinine. The fluoridation of salt or water was considered ideal when the excretion factor amounted to 6.29 mumol X mmol-1. The most important finding was that the Glarus females excrete higher levels of fluorine than the patients from Basel, though the difference was no significant. The fluoridation of salt with 90 mg F/kg is followed by an increase of the excretion factor from 2.58 to 3.65 mumol X mmol-1. It could also be demonstrated that in Canton Glarus, where salt with higher fluorine content is used, the excretion coefficient remains below the level believed to be toxic in the long run. It is concluded that salt fluoridation with 250 gm F/kg is safe. Furthermore, the excretion of fluorine in the control group seems to confirm that fluorine is a trace element of ubiquitous occurrence even excreted in urine of individuals who deliberately avoid fluorine as an additive to table salt or water.

Myoinositol--a uremic neurotoxin?

In 28 patients predialysis plasma myoinositol was significantly elevated to 10.8 +/- 2.5 versus 0.75 +/- 0.15 mg/100 ml in normals (mean +/- 1 SD), and was weakly correlated with plasma creatinine concentrations (r = 0.42, p less than 0.05). Dialysis decreased plasma myoinositol concentrations to 3.5 +/- 1.1 mg/100 ml. In 16 patients studied more extensively plasma myoinositol was similarly elevated to 10.6 +/- 3.4 mg/100 ml, and spinal fluid myoinositol was increased to 9.4 +/- 2.3 mg/100 ml (versus 2.7 +/- 0.7 in normals, p less than 0.001). Nerve conduction velocities were reduced in 15 and EEG tracings were abnormal in 12 of the 16 patients. However, neither spinal fluid nor plasma myoinositol showed any correlation with nerve conduction velocities or EEG changes (r = 0.05-0.20, NS) Myoinositol is one of the substances retained in renal insufficiency but no indication was found that it is a toxin causing uremic neurological disturbances.

Automated blood count analysis by trimodal size distribution of leukocytes with the SYSMEX E-5000.

The automated haematology analyser, SYSMEX E-5000, measures and computes quantitative haematological parameters, and determines the size distribution of blood cells and platelets. After partial lysis, the analyser classifies the leukocytes into 3 populations: small cells (lymphocytes), intermediate sized cells (basophils, eosinophils, monocytes) and large cells (neutrophils, including band cells). One thousand blood samples from inpatients and outpatients were analysed automatically in the SYSMEX as well as being submitted to microscopic blood smear differentiation, and the results were compared. The trimodal size distribution of the automated analysis revealed 1.8% false normal results. Ten cases of eosinophilia of between 6.6 and 12.5% remained undetected by the automated method, which also failed to detect 7 cases of left shift with normal leukocyte count, as well as a single sample containing 2% of myelocytes. Both diagnostic sensitivity and diagnostic specificity were high, i.e. 97.1% and 81.8%, respectively. The predictive values were also high for both pathological and normal results. Since certain changes in blood cell morphology are not detected by the SYSMEX, certain clinical indications still call for a microscopic blood smear examination. With due regard to these limitations, the apparatus yields reliable results and economizes considerably the routine laboratory work load. In the present study, 31% of the microscopic blood cell differential counts were saved by using the SYSMEX E-5000.