Mira Barak

Phagocytosis-induced mutagenesis in bacteria.

Dark mutants of the luminous bacteria Photobacterium fischeri reverted to hereditary stable luminescent forms when incubated with human polymorphonuclear neutrophils (PMN). The maximal mutagenic effect occurred during the first 15 min of phagocytosis, and was dependent on the phagocyte:bacterium ratio as well as on the integrity of the PMN cells. Heat-killed phagocytes or disintegrated phagocytes did not show any mutagenic activity, whereas the supernatant of the phagocytosis reaction exerted mutagenic activity. Scavengers of hydroxyl radical such as mannitol or benzoate and scavengers of singlet oxygen such as beta-carotene, as well as the presence of superoxide dismutase, prevented the mutations. The role of reactive oxygen metabolites in the phagocyte-mediated mutagenic process is discussed.

Neopterin augmentation of tumor necrosis factor production

Gamma interferon (gamma-IFN), lipopolysaccharide (LPS)-gamma or interleukin-2 (IL-2)-induced tumor necrosis factor alpha (TNF alpha) production by both macrophages and peripheral blood mononuclear cells (PBMC), was increased in the presence of neopterin. Addition of neopterin caused an increased level of TNF alpha, but did not affect the kinetics of the TNF alpha production, which showed peak levels of cytotoxic activity 4 h after stimulatory treatment. Using anticytokine antibodies, we concluded that the neopterin effect was mainly gamma-IFN mediated, and only slightly affected by anti IL-2 receptor antibodies. The neopterin augmented TNF alpha production can be attributed to an immunological role for neopterin in the enhancement of cell-mediated immune (CMI) response.

Binding of Serum Prostate Antigen to Concanavalin A in Patients with Cancer or Hyperplasia of the Prostate

The percentage of nonglycosylated prostate-specific antigen (PSA) was measured in the serum of 15 prostate cancer patients and 15 patients with benign hyperplasia of the prostate. The larger part of serum PSA in both groups was glycosylated, but while in carcinoma of the prostate the mean percentage of nonglycosylated PSA was 38.4 +/- 6.5, in benign prostate hyperplasia (BPH) only a mean of 14.2 +/- 4.3% of the PSA was nonglycosylated. These significantly higher results (p less than 0.001) suggest a different pattern of release of PSA from cancer cells and from hyperplastic or normal cells. Since in a part of the BPH we encounter elevations of PSA similar to the levels found in neoplasms, the degree of concanavalin A binding can provide an additional means in differentiating between benign and malignant lesions.

The effect of immunomodulators on PHA or γ-IFN induced release of neopterin from purified macrophages and peripheral blood mononuclear cells

Immunomodulators cause changes in neopterin-release from purified macrophages or peripheral blood mononuclear cells by affecting the macrophage and T cell subsets activity, the intracellular cGMP/cAMP balance, or the intracellular pteridines-related biochemical pathways. Increased neopterin release was achieved by gamma-IFN or its inducers (PHA, IL-2), by interfering with the biopterin production by increased levels of cGMP or by decreasing the activity of the T suppressor cells. The released neopterin levels decreased due to decreased macrophage and T-helper cell activity or due to increased levels of cAMP. The in vitro effect of the immunomodulators has to be taken into account when assessing the neopterin levels in immunomodulators-treated patients.

Bacterial bioluminescence as an early indication of marine fish spoilage

SummaryThe relationships between different microbiological and biochemical parameters and the development of bacterial luminescence associated with the spoilage of marine fish from the Mediterranean-Sea was studied during storage at different temperatures. The bioluminescence level of the bacterial suspensions that were taken from the fish skin increased during the storage; at 20°–25°C the growth and luminescence of the luminuous bacteria correlated well with the total bacterial count while at 5°C the bacterial proliferation was not accompanied by a parallel increase in luminescence.The shift in storage temperature from 25°C to 5°C stabilized the level of the luminescence of bacterial suspension taken from the winter fish which were comprised mainly by Photobacterium phosphoreum, and caused a drop in the luminescence of bacterial suspension taken from the fish caught in the summer which were comprised mainly by Beneckea barveyi. The increase in the bioluminescence level appeared earlier than the increase in trimethylamine level and occured approximately at the same time as the increase in the hypoxanthine concentration. The potential value of the use of bacterial bioluminescence as an early indication for marine fish spoilage is discussed.

The induction of bacterial bioluminescence system on solid medium

Induction of the luminescence system of the marine luminous bacteriumVibrio harveyi growing on a solid medium was studied. The induction occurred when the total number of cells per square centimeter of solid medium approached 104 cells (i.e., either 100 colonies consisting of 100 cells per each colony or 1 colony with about 104 cells). The preinduction period and number of cells per colony at the time of induction decreased as the number of colonies per cm2 increased. The ecological significance of the induction of the luminescence system on solid medium is discussed.

The use of luminous bacteria for determination of phagocytosis

The existing methods for phagocytosis evaluation are inadequate for assessing all the real events occurring during phagocytosis, or for continuously following the kinetics of the process. Our purpose is to establish the use of luminous bacteria as an object for phagocytosis. The bioluminescence test offers an easy and simple method to determine the kinetics of phagocytosis by following the luminescence of the bacteria. The terrestrial luminous bacteria Vibrio cholerae var. albensis are readily phagocytosed by polymorphonuclear (PMN) cells. The correlation coefficient between the decrease in luminescence and the decrease in viable count is 0.999. The rate of decrease in luminescence and the residual level of luminescence after 60 min of phagocytosis are proportional to the rate of increase in phagocytosis induced chemiluminescence, and to its maximal level, respectively. Opsonization requirements are comparable in both tests. Different inhibitors of the phagocytosis process caused similar changes in the rates of the bio- and chemiluminescence (correlation coefficient 0.974), and in the luminescence maximal level (correlation coefficient 0.804). The validity of the bioluminescence assay being proved, it is suggested as an alternative assay for phagocytosis assessment.