Lipid-bound sialic acid, prostaglandin E and histamine in head and neck cancer.

Blood concentration of lipid-bound sialic acid (LBSA), prostaglandin E (PGE) and histamine were determined in 37 patients with carcinoma of hypopharynx and larynx (supraglottic and glottic), in 12 non-cancer patients and in 10 healthy subjects. The concentration of LBSA was significantly increased in 94.4% cancer patients preoperatively and fell to somewhat lower levels within 1 month after tumour resection. In patients with complete tumour resection and no tumour recurrences within 2 years, it steadily decreased thereafter, reaching normal levels within 6-24 months after surgery, whereas in patients with tumour recurrences or incomplete tumour resection it rose again within 6 months after tumour resection. Similarly, the concentration of PGE was significantly increased in about two thirds of cancer patients (67.6%) preoperatively, dropped significantly within 1 month after tumour resection and rose again in patients with tumour recurrences. Preoperative histamine concentration was decreased in 24.3% of cancer patients and postoperatively it rose both in patients with or without tumour recurrences.

Membrane modifications of red blood cells in Alzheimer's disease.

Red blood cells (RBC) from 24 Alzheimer's disease (AD) patients, 18 age- and sex-matched nondemented (ND) patients, hospitalized in the same facility for orthopedic problems, and 18 healthy volunteers aged 30-52 years were studied in order to gain insight into the nature of RBC membrane modifications in AD. Significant differences were found between RBC from AD and ND patients or young controls respectively for annexin V-binding (45.5 +/- 18.0% vs 27.1 +/- 14.7 and 2.7 +/- 1.9, p = .003), fraction of glycerol resistant cells (30.8 +/- 11.1% vs 19.6 +/- 6.4 and 10.2 +/- 3.1, p = .026), cell electrophoretic mobility in polymer (1.028 +/- 0.022 microns sec-1 V-1 cm vs 1.046 +/- 0.022 and 1.053 +/- 0.021, p = .02) and only limited significance for the filterability (1.46 +/- 0.12 msec vs 1.58 +/- 0.11 and 1.54 +/- 0.11, p = 0.1). A logistic analysis, using simultaneously several features as independent variables, suggested the combined use of annexinV- binding, glycerol resistance, and cell filterability which allowed the assignment of 95% of patients from this cohort to the right group. A prospective analysis of a larger cohort is required for the estimation of the diagnostic value of this test battery. In addition, the high level of annexin binding is characteristic of a disruption of the phospholipid asymmetry in aged or damaged cells, while the high glycerol resistance combined with low electrophoretic mobility an rigidity characterize young RBC, thus indicating an enhanced turnover of RBC in Alzheimer's disease.

Ketoconazole inhibits acetaminophen-induced hepatotoxicity in mice.

OBJECTIVE: To investigate the effect of ketoconazole on acetaminophen (AAP)-induced hepatotoxicity in mice. MATERIALS AND METHODS: Mice were given AAP intragastrically (300 mg/kg) and treated with ketoconazole (100 mg/kg intraperitoneally) or saline either 30 min before or 2-3 h after AAP administration. Mortality was recorded for 48 h, during which all mice given saline either died or recovered fully. Serum alanine and aspartate transaminase levels were determined 24 h after administration of AAP. Prostaglandin E2, thromboxane A2 and leukotriene C4 production was determined 6 h after AAP administration in the supernatants from the short-term culture of liver fragments by radioimmunoassay. RESULTS: Ketoconazole significantly decreased mortality and transaminase levels when given to mice either 30 min before or 2 h after AAP. Liver fragments from mice with AAP hepatitis produced greater quantities of prostaglandin E2, thromboxane A2 and leukotriene C4 than fragments from normal liver. Pretreatment of mice with ketoconazole or its addition to liver fragments ex vivo further increased the production of prostaglandin E2 and reduced the production of thromboxane A2. The effect of ketoconazole on leukotriene C4 synthesis was different in vivo (synthesis stimulation) from in vitro (synthesis inhibition). CONCLUSION: The protective effect of ketoconazole in AAP hepatitis is most probably mediated by modulation of eicosanoid synthesis by liver cells.

Effect of fatty acid treatment in cerebral malaria-susceptible and nonsusceptible strains of mice.

Cerebral malaria-susceptible (C57BL/6) mice infected with Plasmodium berghei ANKA (PbA) developed low parasitemia and died from typical neurological symptoms between 8 to 10 days after infection. In contrast, nonsusceptible (BALB/c) mice developed high peripheral blood parasitemia and died 22-24 days later without neurological implications. Daily injections of fatty acids (FA) during the first 3 days after infection protected C57BL/6 mice from cerebral symptoms but had no effect on BALB/c mice. Thus, treated C57BL/6 mice developed hyperparasitemia and died 25 days after infection, like BALB/c mice. Red blood cells from C57BL/6 control mice were found to be more resistant to lysis by linoleic acid than those of BALB/c mice. Three days following infection with PbA, these differences disappeared. Treatment with FA prevented these changes. We concluded that the host's cells were altered soon after infection and that the nature and degree of alterations depended on the mouse strain, thus determining the eventual outcome of the infection. Likewise, the effects of FA might not be directed against the parasite but rather seem to act early after infection on these parasite-induced modifications of host cells.

[Histamine levels in the blood in patients with malignant tumors]. / Razina histamina u krvi kod bolesnika s malignim tumorom.

The concentration of histamine in blood was determined in 22 patients with solid malignant tumors, 16 hospitalized non-cancer patients and 9 healthy subjects. Patients with cancer (mainly carcinomas of the breast and gastrointestinal tract) were divided into two groups: patients with resected primary tumor without known metastases (group I) and patients with present primary tumor with or without metastases (group II). In comparison with the healthy subjects (histamine concentration 69.0 +/- 6.03 ng.ml-1), cancer patients in both groups had significantly decreased levels of histamine in blood. Also, the concentration of histamine in patients with present tumor (group II; 40.1 +/- 3.48 ng.ml-1) vas significantly lower than in patients with resected primary tumor (49.9 +/- 3.14 ng.ml-1). Furthermore, hospitalized non-cancer patients had lower, but not statistically significant, concentration of histamine (59.7 +/- 6.13 ng.ml-1) than healthy subjects. These findings, together with similar data of other authors, suggest that decreased level of histamine in blood might be a good nonspecific marker for onset and progression of solid malignant tumors.

T and B origin of human lymphocytic leukaemia.

A new immunological classification of leukaemia diseases in man was assessed by using electrophoretic mobility as a cell-marker. It was found that all acute lymphocytic leukaemia analysed in childhood were T-cell leukaemia, and the majority of chronic lymphocytic leukaemia in adults were also T-cell leukaemia. The remainder, chronic lymphocytic leukaemia in adults, were B-cell leukaemia. The above statements are only partially compatible with the classification given by the most commonly used T- and B-cell markers such as rosette formation or cell-surface immunoglobulins.

Immunopharmacological properties of a protein-bound histamine metabolite.

Histamine metabolite was prepared by incubating histamine dihydrochloride and diamino oxidase for 24 h at 37 degrees C. Radioactive histamine was used for monitoring the whole procedure and to select the best experimental protocol. Pharmacological activities of histamine are abolished by this procedure. Histamine metabolite was found to bind to the serum proteins, to inhibit the PHA response of mouse and human mononuclear cells and to accelerate mortality rates in tumor-bearing mice. Thin layer chromatography allowed separation of metabolite from histamine and from known imidazol-derived compounds. This is the first experimental evidence for immunological properties ascribed to a histamine metabolite.

Blood compatibility of particulate carriers investigated by cell electrophoresis and particle electrophoresis: a preliminary report.

Polyethylene glycol (PEG) and dextran were covalently coupled, or only adsorbed, to the surface of three kinds of inorganic particles in order to shield their surface and reduce their nonspecific binding to red blood cells. Surface modification as well as interaction of particles with red blood cells was followed up by particle electrophoresis. This allowed a quick evaluation of the efficiency of polymer coupling. Moreover, the nonspecific binding of particles to red blood cells was easily investigated with cell electrophoresis, showing the inhibitory effect of immobilized PEG-5000 or dextran. The electrophoretic mobility analysis presented here may be used for screening blood compatibility of particulate drug carriers and could be helpful in formulating long-living circulating particles.

Further studies on the H-2 linked dependence of the adjuvant action of Brucella abortus.

The B 19S strain of Brucella abortus is found to act as an adjuvant to the anti-sheep red blood cell (SRBC) reaction in some congenic strains of mice but not in others. If the recipient is H-2b, there is no adjuvant effect (B 19S); neither is there (thymus-dependent anti-B 19S reaction as measured by thymocyte activation and change of electrophoretic mobility. In contrast, there is a thymus-independent anti-B 19S reaction (production of haemagglutinins) as good in the H-2b mice as in the others. Experiments with T cell deprived mice show that the adjuvant action of B 19S is thymus-dependent. As the anti-SRBC reaction without adjuvant is also thymus-dependent, it is difficult to distinguish anti-SRBC and anti-B 19S reactions from the adjuvant action of B 19S on the anti-SRBC reaction. Several explanations are possible, all involving H-2 (Ir?) controlled thymus dependent mechanisms.

Particle electrophoresis of micrometric-sized superparamagnetic particles designed for magnetic purification of cells.

Measuring the electrophoretic mobility of superparamagnetic particles (0.5-4.5 microns mean size) was undertaken to probe the coupling of lectins and antibodies to their surface. Coupling was either noncovalent (antigen-antibody and biotin-streptavidin linkage) or covalent (tosyl-activated beads). The direct observation of the electrophoresis of single particles illuminated in dark field and processed by image analysis allowed the determination of their apparent electrophoretic mobility. Mobilities ranged from -0.5 micron s-1/CmV-1 to +1 micron s-1/CmV-1 when measured at 20 degrees C in 0.15 M NaCl and 30 mg/mL sorbitol, pH 7.4. The relative standard deviation was less than 0.1%. Surface immobilization of charged proteins onto the superparamagnetic beads shifted their electrophoretic mobility up to 200%; this was also quantitatively correlated with some specific properties (enzymatic activity, antigen-binding activity, lectin-binding activity). Although particle electrophoresis has mainly been reported for the study of surface adsorption phenomenon, it may be a versatile tool for controlling covalent modifications of particles designed for therapeutical targeting or chromatographic use and may also apply to a quantitative analysis of ligand-binding interactions.

Characterization of a subpopulation of mouse red blood cells as preferential target for malarial invasion.

Cell electrophoresis was used to study a distinct subpopulation of murine red blood cells (RBC). These RBC are normally found in the spleen and bone marrow. They appear in the peripheral blood when mice are mildly bled or sucked by mosquitos. These cells have been characterized as having larger size, light density, lower electrophoretic mobility, and being more resistant to lysis with unsaturated fatty acids and in glycerol-containing medium than mature erythrocytes. All their features suggest that their differentiation status represents an intermediate stage between reticulocytes and adult RBC. In vitro Plasmodium invasion tests showed their increased sensitivity to invasion by the parasites. The extent of their spreading in the blood was found to be strain-dependent, being more pronounced in C57B1/6 mice, highly susceptible to developing cerebral malaria after infection with Plasmodium berghei, as compared to Balb/c, a nonsusceptible strain of mice.

Influence of milk source on transplantability of histocompatible mammary tumours in mice.

It is confirmed that C3H mammary tumours are much more easily transplantable in histocompatible recipients when these have been reared on C3H milk, than when they have been reared on milk from the inbred Swiss/B strain. By contrast, A.CA mammary tumours transplanted in histocompatible hosts reared on A.CA or Swiss/B milk, grow almost equally well in both sorts of recipient. Thus, rearing on Swiss/B milk has different effects on the transplantability of mammary tumours of C3H and A.CA. On the other hand, recipients which were reared on C3H or A.CA milks accept grafts of C3H mammary tumours about equally, suggesting that milks from A.CA and C3H have the same effect on the transplantability of C3H mammary tumours. The different action of Swiss/B milk on tumours of C3H and A.CA seems best attributed to differences between C3H and A.CA tumours or between mouse strain genotypes. By contrast, the transplantability of C3H mammary tumours is significantly changed when the recipients were reared on milk from the RIII strain instead of C3H. These facts suggest that the milk from RIII has an action which differs from that of both C3H and A.CA in this respect. The data are discussed on the basis of a differential tollerance-inducing action of mammary tumour viruses (MTVs) which infect C3H, A.CA and RIII, and have an important role in tumour induction.

Effect of a protein bound polysaccharide (PS-K) on tumor development and infections in splenectomized rats and mice.

Daily oral administration of PS-K, an immunomodulating agent, was found to improve survival rate in tumor bearing mice and rats and to decrease susceptibility to infection in mice. Surprisingly, splenectomized tumor bearing rats and mice responded better to PS-K treatment as measured by survival rate, tumor size and immunosuppressive properties of serum. The mechanism of PS-K antitumor activity in both experimental systems might be due to the alteration of splenic immune suppressive mechanisms during tumor growth in the host. On the other hand, host resistance to acute infection was found to be less dependent upon the presence or absence of spleen suggesting a different mechanism of PS-K activity in acute infectious disease.

Cellular reaction induced by "nonantigenic" substances.

A previously designed isotopic in vivo method for measuring delayed hypersensitivity was used with nonantigenic (or weakly antigenic) phlogogenic substances. A strong reaction was observed in some circumstances, witnessing the fact that the method has a specific and a nonspecific component. The reaction is drastically augmented by a mild whole-body irradiation of the reacting animals. In the case of the studied substances (carbon, oil and bentonite) the reaction is bound by polymorphonuclear neutrophils (PMN), but it was found to be independent of the number of circulating PMN in the blood.

Spontaneous maze ambulation in two mouse strains.

A simple device is described, which permits us to quantify several parameters of spontaneous behaviour of small animals. Using this device with mice we obtained statistically satisfactory results, showing a strong genetic influence on the behavioural characteristics tested.

Cyclophosphamide-induced suppressor cells in nude mice.

Lymphocytes regenerated after treatment with a high dose of cyclophosphamide (CY) were characterized in nude mice. Ten days after a single injection of 200 mg/kg CY into nude mice, regenerated spleen cells suppressed in vitro primary and second antibody production against sheep red blood cells. The CY-treated spleen cells exhibited normal natural killer (NK) activity, very low B and T cell content, but increases in cell surface charge [electrophoretic mobility (EPM)] and histamine receptors. The suppressor cells could not be removed by treatment with anti-Thy-1 plus complement (C), or treatment with antiasialo GM1 (aGM1) plus C, which abrogated NK activity. It was concluded that CY-treated spleen cells, which exhibited high EPM and histamine receptors, comprise the natural suppressor cells which are Ig-, Thy-1- and aGM-1.

The action of various sera on colony formation and DNA synthesis of murine thymocytes in culture.

Murine thymus cells were caltivated up to 10 days in a semi-solid medium. Both macroscopically visible colonies and DNA synthesis (identified by 3H-thymidine incorporation) were counted with and without added sera. According to their origins the sera exerted different modifications of the cultivated cells behaviour. Changes of colony number and of thymidine incorporation were quite independent on each other.

Blood lymphocytes in acute lymphoid leukaemia in remission and in relapse. Predictive value of electrophoretic mobility and refringence.

Blood lymphocytes were studied during the evolution of acute lymphocytic leukaemia, including remissions and relapses, under chemotherapy and immunotherapy. Lymphoblasts migrate as a uniform population and have a mean electrophoretic mobility of 1.04 mum-sec-1. V-1cm. These lymphoblasts are also refringent. The relapse seems to be preceded by a slowing of the electrophoretic mobility of fast cells and a rise in the percentage of refringent lymphocytes.

T- and B-cell surface markers on rabbit lymphocytes.

Rabbit thymus, appendix, blood and lymph nodes were characterized using immunological tests: rosette-forming ability with homologous or heterologous red blood cells, surface immunoglobulins, analytical cell-electrophoresis, cell refringency and in vitro mitogen responsiveness. The experiments were conducted on normal rabbits and cyclophosphamide-treated rabbits. Moreover, we tried to separate slow and high mobility cells by free-flow electrophoresis into fractions. These experiments suggest that rabbit lymphocyte behaviour is exceptional among mammals in that the majority of their lymphocytes appeared as B cells and that, in comparison with mouse and man, the proportions of these cells are inversed.