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A haematological and biochemical investigation was undertaken in 122 consecutive male patients with suspected organic solvent poisoning due to exposure to a mixture of organic solvents such as turpentine, toluene and xylene. Sixty-four healthy solvent exposed and 91 healthy non-exposed male volunteers were used as controls. The only statistically significant differences were that in the patients (B)-leucocytes and S-creatinine were lower than in the controls. (B)-monocytes were higher in all exposed groups than in the controls. S-creatine kinase was higher in patients actually exposed at the examination time, than in all other groups. B-hemoglobin, B-erythrocytes, (B)-Erc-reticulocytes, (B)-hematocrit, (B)-MCHC, (B)-MCV, (B)-Lkcs-differential count except monocytes, (B)-thrombocytes, B-sedimentation rate, P-orosomucoid, P-immunoglobulins (IgG, IgA and IgM), S-alanine amino transferase, S-urate, U-glucose, U-albumin and U-hemoglobin were unchanged. It is concluded that there was no characteristic haematological and biochemical pattern that could be of value in the individual diagnosis of organic solvent poisoning. The patients and the controls were seen as out-patients. In all groups studied, more than 10% of the results of reticulocytes, leucocytes, sedimentation rate, orosomucoid and creatine kinase exceeded the upper level of the reference interval, which is based on in-patients. New reference intervals for these 5 analyses, valid for ambulantly examined subjects, should be worked out.

We have used human male-specific 3.4 kb Hae III restriction endonuclease fragments to explore the evolutionary history of man's Y-chromosome. We have identified four sets of reiterated sequences on the basis of their relative sequence homology with autosomal DNA. The sequences account for approximately 40% of the human Y-chromosome, are interspersed within the same 3.4 kb Hae III fragments, are heterogeneous and contain all reiterated DNA previously demonstrated to be specific for the Y-chromosome (it-Y DNA). Y-specific 3.4 kb Hae III sequences do not reassociate with either human female or ape DNA at standard reassociation criteria. However, approximately half of it-Y DNA (cross reacting it-Y) reassociates with both human female and ape DNA at reduced reassociation criteria. The remaining half (Y-specific it-Y) retains its specificity for the human Y-chromosome. These two sets of it-Y DNA have distinct reiteration frequencies and thermal stabilities with their Y-chromosome homologs. Non-Y-specific 3.4 kb Hae III sequences reassociate with both human female and ape DNA at standard reassociation criteria. The abundance of these non-Y-specific sequences decreases as a function of their evolutionary distance for man. One subset of non-Y-specific 3.4 kb Hae III sequences forms stable duplexes with human Y-chromosome DNA and with human and ape autosomal DNA. No detectable base-mismatch occurs among these homologs suggesting complete conservation of these sequences during primate evolution. The second subset of Non-Y-specific Hae III sequences form stable duplexes with human Y-chromosome DNA but highly mismatched duplexes with human and ape autosomal DNA. The finding that homologs of 3.4 kb Hae III sequences are not found within the Y-chromosome of apes but are only present in autosomes suggests that 3.4 kb Hae III sequences are largely autosomal in origin. Since autosomal homologs of most 3.4 kb Hae III-sequences exhibit a greater degree of divergence that those localized to the Y-chromosome, their evolutionary history seems to be chromosome-dependent. Our findings are not easily correlated with the comparative morphology of primate Y-chromosomes and suggest that sequence rearrangement has been a major event in the evolution of the human Y-chromosome. The significance of the specific interspersion of four sets of reiterated sequences, with distinct evolutionary histories, within a repeating unit specific to the human Y-chromosome is not clear. The apparent conservation of at least some of these reiterated sequences suggests they may be of functional importance.

Enzymuria, urinary cell excretion and proteinuria were simultaneously determined in renal diseases of female Wistar rats in order to investigate the diagnostic value of urinary enzymes. Investigations were carried out on rats with E. coli-pyelonephritis facilitated by oestradiolundecylate, aminonucleoside nephrosis, aminoglycoside induced renal lesions and pyelonephritic animals treated with therapeutic and toxic doses of tobramycin. --From the results of these studies it was concluded that the main diagnostic value of urinary enzymes is detection of drug induced tubular lesions in individuals with preexisting renal diseases.

Blood cells proliferate extravascularly in the bone marrow and enter the circulation by migrating through endothelial cells of venous blood sinuses. This migration, or diapedesis, was suspected to involve actin. To test for the presence and distribution of actin, sections of rat bone marrow were examined by indirect immunocyto-chemistry. Affinity purified rabbit antichicken gizzard actin antibody, and goat-antirabbit IgG-FITC, or goat antirabbit IgG colloidal gold probes were used. The migrating cell contacts the endothelial cell and forms a podosome (a cortical bleb). Immunocytochemistry shows this region to contain actin. As diapedesis proceeds the podosome deforms, then breaches the endothelial cell. At this time the anterior portion of the leukocyte shows heavy labeling for actin. When the migratory cell traverses approximately half of its length through the endothelial cell, actin appears prominent in the caudal region of the cell. The immunocytochemical data suggest that actin is nonrandomly distributed in leukocytes undergoing diapedesis and may be a component of the force-generating mechanism responsible for this transcellular migratory event.

Cytosol proteins from normal lymphocytes, leukemic lymphocytes, and different cultured lymphoid cell lines were separated by two-dimensional mini gel electrophoresis. By staining with Coomassie blue, specific protein patterns were obtained. Very similar gel maps were produced by the cytosol proteins of chronic lymphocytic leukemia cells, hairy cells, and of in vitro grown B cells. Protein 36/6.2 (molecular weight/isoelectric point), consistently present in these cells, could not be demonstrated in normal lymphocytes. For the comparison of control Raji cells--an Epstein-Barr-Virus (EBV)-DNA carrying Burkitt's lymphoma cell line--with Raji cells induced for early antigen (EA) production, 35S-methionine-labelled total cell lysates were analyzed. In the induced cells, an additional protein (100/5.5) was found; this might be one of the immunologically defined EBV-associated antigens. These results demonstrate that two-dimensional mini gel electrophoresis can be useful for the characterization of leukemic cells in addition to the morphological, cytochemical, and surface marker analyses.

Using pulse cytophotometry, almost quantitative separation of the leucocyte fraction from DNA histograms was possible by means of an anticoincidence discrimination device, This modified technique was employed for biparametric DNA/protein measurements of voided urine samples, bladder washings, and tumour tissues. The results show a high degree of correlation between these samples so that, for tumour diagnosis from DNA histograms, voided urine specimens can be used rather than bladder washings. The criteria for the bladder tumour diagnosis are derived from DNA measurements of 32 controls and 35 tumour patients. The diagnostic sensitivity of this method is 0.91 and the specificity 0.75.

A selective staining of hemoglobin in erythroid cell series was achieved by use of Sudan Black B (modified method of Sheehan and Storey) if optimal amount of hydrogen peroxide was added to the staining mixture. The effect of some inhibitory agents (KCN, wet heat, pH) on this staining as well as on the Lepehne's pseudoperoxidase reaction for hemoglobin was similar. Both reactions were more resistant to these factors than the peroxidase reactions and sudanophilia in granulocytes in which both could be blocked by the pretreatment with absolute methanol. Moreover the effect of some extraction procedures for lipids on both myeloperoxidase reactions and sudanophilia was investigated. The results support the view that the sudanophilia in granulocytes is due to their peroxidase activity and for the staining of hemoglobin by use of Sudan Black B with H2O2 its pseudoperoxidase activity is responsible. In addition the effect of the substitution of phenolphosphate by dihydroxybenzenes on granulocyte sudanophilia is reported.

Evidence on the action mechanisms of gold salts in the treatment of rheumatoid arthritis is still inconclusive. The intracellular localization of the place of action is likely. Therefore not only the serum gold levels but also the intracellular concentration of gold are of special interest. We measured the gold concentration in the serum and in the blood cells after in vitro application of aurothiomalate (Tauredon), gold keratinate (Auro-Detoxin) and triethylphosphine-gold (Ridaura) and in blood samples of patients undergoing these gold salts treatments. Cell-bound concentrations were found to vary extensively as a function of the gold compound used. While no or very little gold was present intracellularly after administration of the 2 parenteral drugs, up to 40% of the circulating gold was found to bind to the cells after administration of the triethylphosphine compound for gastro-intestinal absorption. The red cell concentration was more or less the same as that in the extracellular compartment. Gold apparently accumulated in the white cells, because the cell-bound concentration relative to unit volume was up to 20 times higher than the plasma level. The method used did not offer any information on the actual binding site of gold in white cells, i.e. cytoplasm versus nucleus versus cell membrane.

A recently described immunoperoxidase method for the detection of nuclear human cytomegalovirus (HCMV) immediate early antigen (IEA) directly on peripheral blood leucocytes suffers from the drawback that the antigen is vulnerable to endogenous peroxidase inactivation procedures. To solve this problem a procedure is developed in which endogenous peroxidase is inactivated after binding and immobilization of the primary antibody with 4% formaldehyde. In combination with this procedure, three types of inactivation were investigated: glucose/glucose oxidase, hydrochloric acid and methanol/H2O2. Of these three, the first gives optimal results, especially in combination with methanol/acetic acid (20/1 v/v) as the primary fixative. This procedure results in preparations which allow for a more objective evaluation and enable automated examination using bright field microscopy. As a second improvement we developed a simple adherence method in order to diminish the risk of infection for the laboratory staff during processing of unknown blood samples. The protocol described shows great clinical potential for the diagnosis of HCMV infections.

A small unconventional, viroid-like aetiological agent might initiate Crohn's disease (CD). Electrophoresis of radiolabelled low molecular weight nucleic acids from mesenteric lymph nodes and leucocytes did not distinguish CD-specific sequences compatible with a viroid-like agent.

The localization of ferritin was studied in peripheral blood cells and variously fixed tissues with the antibodies against ferritins isolated from human heart and spleen. The unlabelled antibody enzyme method (PAP) was used to detect the binding sites of antibodies. In peripheral blood cell smears both antisera gave rise to strong staining of polymorphonuclear (PMN) cell cytoplasm, whereas the monocytes stained relatively weakly. There were no staining differences between the two antisera. In human spleen sections the spleen ferritin antiserum stained the PMN cells and sinusoidal lining cells, whereas the heart ferritin antiserum stained only PMN cells. Neither of the two antisera stained monocytes in the spleen sections. This finding was observed in specimens fixed in Bouin's fixative, Baker's fixative and neutral formalin. However, the immunoreactivity of ferritin was totally destroyed by some other fixatives (Carnoy's fixative, formol sucrose and glutaraldehyde). These results suggest that ferritin is more readily released from monocytes than from PMN cells, and that mature spleen macrophages contain antigenic determinants of ferritin that are recognized only by anti-spleen ferritin antiserum.

Human leukocyte-derived alpha interferon [HuIFN-alpha(Le)] has been purified and/or concentrated on Carboxymethyl derivatized Controlled Pore Glass (CML-CPG240) beads. These glass beads adsorb HuIFN-alpha(Le) efficiently at acid pH and at physiological ionic strengths. Elution of HuIFN-alpha(Le) may be accomplished by several methods. Using buffers at relatively high ionic strengths (approximately 0.6 M) and pH values ranging from 2.6 to 6.9 for elution, preparations with specific activities of 10(5)-10(6) IU/mg were obtained with approximately 90 percent recoveries. Alternatively, using elution buffers at the same high ionic strength and at pH values ranging from 7.0 to 8.0, five-fold or better concentration and complete recovery of crude HuIFN-alpha(Le) were achieved. The resulting preparations were suitable for direct application to an antibody affinity chromatography column.

A technique is described which permits the direct, infrared (IR) spectrophotometric observation of peripheral leukocytes by utilizing multiple internal reflectance (MIR) spectroscopy on zinc selenide prisms. The IR spectra of nitro-blue tetrazolium (NBT) and its derivative, nitro-blue diformazan (NBF), are described and examined and these spectra are compared with those of the intracellular NBT and NBF yielding several peaks which may prove to be analytically useful. Additionally, the presence of unreduced NBT in leukocytes is demonstrated, which microscopic and chemical "NBT tests" have not previously done.