[Molecular mechanisms of lung cancer development at its different stages in nuclear industry workers].

OBJECTIVE: to assess mutational events in exons 5, 7, and 8 of the p53 gene and to reveal mutant p53 protein in verified cases of morphologically altered (proliferative and precancerous changes, lung cancer) and histologically unaltered, lung tissues in workers exposed to occupational radiation. MATERIAL AND METHODS: The investigation used formalin-fixed paraffin-embedded unaltered and altered lung tissue blocks (FFPBs) obtained from the human radiobiological tissue repository. The shelf-life of FFPBs was 5-31 years. An immunohistochemical technique using mouse antibodies against p53 protein (<>, Denmark), stained with diaminobenzidine (DAB) chromogen, was employed to determine p53 protein. DNA was isolated from lung tissue FFPBs with QIAmp DNA FFPE Tissue Kit, (<>, USA). Polymerase chain reaction (PCR) was performed to amplify the p53 gene exons 5, 7, and 8 selected for examination, by applying the sequences of genes and primers, the specificity of which was checked using the online resource (http://www.ncbi.nlm.nih.gov/blast). PCR products were detected by temporal temperature gradient gel-electrophoresis and the Sanger sequencing method. The obtained DNA fragments were analyzed on a sequencer ABI Prism 3100 Genetic Analizer (<>, USA). Computer-aided DNA analysis was made using the BLAST program. A package of applied Statistica 6.0 programs was employed for statistical data processing. Results. Immunohistochemical analysis showed that mutant p53 protein was absent in the cells of unaltered lung tissue and the number of cells with mutant p53 protein increased in all the patients with proliferative and precancerous changes and lung cancer, suggesting p53 protein dysfunction. The total number of p53 gene mutations in exons 5, 7, and 8, if there were proliferative and precancerous lung tissue changes and lung cancer, were 25, 20, and 40%, respectively. All the found mutations were transversions (the substitution of purine for pyrimidine or, conversely), indicating the action of exogenous mutagens. CONCLUSION: The results of this investigation have confirmed other investigators' data showing that p53 gene mutations in lung cancer are observed in 40-70% of cases. The differences in the number of cases of altered lung tissue with mutations in the p53 gene (not more than 40%) and in those of p53 protein expression were found in 100%, suggesting the regulation of p53 gene function in the cell at multiple levels.

[Possibility of DNA isolation from archived autopsy tissues for molecular genetic studies].

Optimal conditions were defined for DNA isolation from formalin-fixed and paraffin-embedded (FFPE) archived autopsy lung tissue slices. The quality of DNA preparations isolated from the FFPE archived slices was assessed and their suitability for further molecular genetic analysis estimated. DNA isolated from the FFPE slices stored less and more than 10 years was suitable for molecular genetic studies in 100 and 66.7% of cases, respectively.

[Radiobiological Human Tissue repository: progress and perspectives for solving the problems of radiation safety and health protection of personnel and population].

Radiobiological Human Tissue repository was established in order to obtain and store biological material from Mayak PA workers occupationally exposed to ionizing (α- and/or γ-) radiation in a wide dose range, from the residents exposed to long term radiation due to radiation accidents and transfer of the samples to scientists for the purpose of studying the effects of radiation for people and their offspring. The accumulated biomaterial is the informational and research potential that form the basis for the work of the scientists in different spheres of biology and medicine. The repository comprises 5 sections: tumor and non-tumor tissues obtained in the course of autopsies, biopsies, surgeries, samples of blood and its components, of DNA, induced sputum, saliva, and other from people exposed or unexposed (control) to radiation. The biomaterial is stored in formalin, in paraffin blocks, slides, as well as in the freezers under low temperatures. All the information on the samples and the registrants (medical, dosimetry, demographic, and occupational data) was obtained and entered into the electronic database. A constantly updated website of the repository was developed in order to provide a possibility to get acquainted with the material and proceed with application for biosamples for scientists from Russia and abroad. Some data obtained in the course of scientific research works on the basis of the biomaterial from the Repository are briefly introduced in the review.

X-ray fluorescence microscopy for investigation of archival tissues.

Several recent efforts in the radiation biology community worldwide have amassed records and archival tissues from animals exposed to different radionuclides and external beam irradiation. In most cases, these samples come from lifelong studies on large animal populations conducted in national laboratories and equivalent institutions throughout Europe, North America, and Japan. While many of these tissues were used for histopathological analyses, much more information may still be obtained from these samples. A new technique suitable for imaging of these tissues is x-ray fluorescence microscopy (XFM). Following development of third generation synchrotrons, XFM has emerged as an ideal technique for the study of metal content, speciation, and localization in cells, tissues, and organs. Here the authors review some of the recent XFM literature pertinent to tissue sample studies and present examples of XFM data obtained from tissue sections of beagle dog samples, which show that the quality of archival tissues allows XFM investigation.

[The significance of immunity disorders in the development of tumorous effects during the long-term action of tritium in mice]. / Znachenie narushenii immuniteta v razvitii opukholevykh éffektov pri dlitel'nom deistvii tritiia u myshei.

In experiments with CBA mice it was shown that the long-term influence of tritium oxide administered with drinking water daily during 180 days (370 kBq.g-1 of body mass, cumulative dose 8.7 Gy, dose rate 4.5 cGy.day-1) causes the development of immunity deficiency 90 days after the onset of the administration that persists for up to 270 days. There is a 34% decrease in the average life of irradiated animals and an increase in the number of malignant neoplasms (postmortem examination was performed after natural death or killing on days 250, 350 and 450). There is a direct relationship between the occurrence of malignant neoplasms and the immunity deficiency.

[An increase in the incidence of tritium-induced leukoses with the repeated administration of a bacterial lipopolysaccharide]. / Povyshenie chastoty indutsirovannykh tritiem leikozov pri povtornom vvedenii bakterial'nogo lipopolisakharida.

The incidence of leukoses upon long-term administration of tritium oxide to Wistar rats increased by 15.6% as compared to 2.1% in controls. The repeated injection of bacterial lipopolysaccharide caused a 2.3-fold increase in the occurrence of hemoblastosis induced by the radionuclide mainly due to the increase in the proliferative activity of haemopoietic precursors because of the increased demand for mature cells.

[Cytogenetic characteristics of H3 oxide induced myeloid leukemia in Wistar rats]. / Tsitogeneticheskaia kharakteristika mieloidnykh leikozov, indutsirovannykh okis'iu 3H u krys Wistar.

Frequency and spectrum of chromosomal aberrations were studied in hemopoietic cells of Wistar rats myeloid leukaemia induced by the prolonged exposure to tritium oxide. Differences in quantitative and structural damages of chromosomes were revealed by the cytogenetic assessment of eighteen rats with acute myeloid leukaemia (AML) and six rats with chronic myeloid leukaemia (CML). The most characteristic chromosomal aberrations for AML appeared to be rearrangements in chromosomes 1-3. In CML translocations by one of chromosomes 15-20 coupled with defects in the groups of chromosomes 5-10 seemed to be nonrandom.

[Hematopoietic precursor cells in Wistar rats with acute myeloblastic leukemia]. / Krovetvornye kletki-predshestvenniki u krys porody Wistar s ostrym mieloblastnym leikozom.

The studies were performed on Wistar rats with acute myeloblastic leukemia induced by a prolonged administration of tritiated water (370 MBq/kg/day-1; an absorbed dose--25.3 Gy) and by repeated pyrogenal injections. A reduction of the pool of polypotent hemopoietic cells (CFUs) with a simultaneous increase of their proliferative activity and a decline of their ability to self-supporting were observed in the leukemia-bearing animals. Populations of CFUdc in DC and of CFUc were reduced in all cases; their colony-forming ability was also found to decrease. All animals with leukemia showed a lowered amount of CFU-BL in the bone marrow and of CFU-TL in the thymus, although the content of these cells in the peripheral blood remained unchanged.

[Frequency of structural chromosome aberrations in the hepatocytes of rats exposed to polymeric 239Pu]. / Chastota strukturnykh povrezhdenii khromosom gepatitsitov krys pri deistvii polimernogo 239Pu.

Intravenous injection of polymeric 239Pu(IV) nitrate (166.5, 55.5 and 18.5 kBq/kg body mass) to Wistar rats was shown to produce biphase changes in the frequency of hepatocyte chromosome aberrations. The increase in the structural damages to chromosomes at later times of observation was a pronounced function of radiation dose. The absence of such a dependence at early times was evidently due to the elimination of damaged liver parenchyma cells.

[Comparative evaluation of the carcinogenic effects of chronic exposure to tritium oxide and external gamma-radiation]. / Sravnitel'naia otsenka blastomogennogo éffekta khronicheskogo deistviia okisi tritiia i vneshnego gamma-oblucheniia.

In experiments of on rats a study was made of late effects of chronic action of tritium oxide (3HOH) during 6 months (37 X 10(4) Bq/g/day) and external gamma-radiation (137Cs) which were delivered in comparable daily and cumulative doses. It was shown that 3HOH produced a more pronounced blastomogenic effect. The RBE coefficient of tritium oxide approximated 1, with a reference to the average life shortening, and 4.2 and 2.5, with a reference to the incidence of malignant tumors and leukoses, respectively.