IMPACT OF ASTAXANTHIN ON MANIFESTATIONS OF THE DIRECT AND RESCUE TUMOR-INDUCED BYSTANDER EFFECT.

Objective: to investigate the reciprocal impact on the genome of malignant and normal human peripheral bloodlymphocytes under their co-culture and the possibility to modify the effects by astaxanthin.

Methods: Separate and joint/separate culturing of peripheral blood lymphocytes (PBL) of the chronic lymphocyticleukemia (CLL) patients (n = 6) and conditionally healthy individuals (n = 6), Comet assay method, fluorescencemicroscopy with automated software for the analysis of results, statistical methods.

Results: Both direct and rescue tumour-induced bystander effects were observed under the joint/separate culturing of blood lymphocytes of conditionally healthy individuals (the bystander cells) and blood cells from CLL patients(the inducer cells).

ANALYSIS OF DNA METHYLATION CHANGES IN MANIFESTATION OF DIRECT AND RESCUE BYSTANDER EFFECTS.

Objective: to investigate changes in DNA methylation in bystander and inducer cells during the manifestation ofdirect and rescue bystander effects.

Methods: Separate and co-cultivation of peripheral blood lymphocytes (PBL) of 10 conditionally healthy individuals; γ-quantum irradiation (IBL-237C emitter); modified comet electrophoresis method (Comet assay) under neutralconditions using the methylation-sensitive restriction enzyme HpaII; fluorescence microscopy with an automatedcomputer software system for analyzing the results; statistical methods.

Results: The level of DNA methylation in PBL was quantitatively assessed using DNA migration parameters inagarose gel: the length of the comet tail (in μm), the percentage of DNA in the tail part of the comet, and TailMoment (TM), which simultaneously takes into account both the amount of DNA in the tail part of the comet andthe length of the tail.

MODIFICATION OF THE TUMOR/INDUCED BYSTANDER EFFECT BY IRRADIATION UNDER COCULTIVATION OF LYMPHOCYTES FROM PATIENTS WITH CHRONIC LYMPHOCYTIC LEUKEMIA AND LYMPHOCYTES FROM HEALTHY DONORS.

Objective: Study the tumor-induced bystander effect of blood cells from chronic lymphocytic leukemia (CLL)patients on non-transformed bystander cells (peripheral blood lymphocytes (PBL) of conditionally healthy individ-uals) and the possibility of its modification after the impact of ionizing radiation.

Materials And Methods: We carried out cocultivation and separate cultivation of blood samples from conditionallyhealthy volunteers and patients with CLL according to our technique. Using the Comet assay, the relative level ofDNA damage was evaluated.

THE IMPACT OF ASTAXANTHIN ON THE LEVEL OF DNA METHYLATION IN IRRADIATED IN VITRO HUMAN LYMPHOCYTES.

Objective: To investigate an effect of astaxanthin on global DNA methylation in human peripheral blood lympho-cytes exposed to γ-radiation in vitro.

Methods: Samples of whole blood were diluted with RPMI-1640 medium in proportion 1 : 10 and incubated at 37 °Cfor 3 h. Samples were exposed to γ-ray (emitter IBL-237C, dose-rate 2.

Study the impact of astaxanthin on developing of genomic instability in human peripheral blood lymphocytes irradiated in vitro on G2 phase of cell cycle.

Objective: To identify the possibility of modification by astaxanthin the level of genome damages induced by gamma quanta in the culture of human peripheral blood lymphocytes exposed in vitro on postsynthetic (G2) phase of the first mitotic cycle.

Materials And Methods: Peripheral blood lymphocytes from four apparently healthy volunteers 35-51 years old were cultivated using modified micromethod. To obtain genomic damages in G2 phase of the first mitotic cycle the part of cultures was irradiated by γ quanta in dose 1.