Analysis of Methylome of Different Forms of Basal Cell Hyperplasia and Squamous Cell Metaplasia of Bronchial Epithelium.

Squamous cell lung cancer (SCLC) occurs as a result of dysregenerative changes in the bronchial epithelium: basal cell hyperplasia (BCH), squamous cell metaplasia (SM), and dysplasia. We previously suggested that combinations of precancerous changes detected in the small bronchi of patients with SCLC may reflect various "scenarios" of the precancerous process: isolated BCH→stopping at the stage of hyperplasia, BCH+SM→progression of hyperplasia into metaplasia, SM+dysplasia→progression of metaplasia into dysplasia. In this study, DNA methylome of various forms of precancerous changes in the bronchial epithelium of SCLC patients was analyzed using the genome-wide bisulfite sequencing. In BCH combined with SM, in contrast to isolated BCH, differentially methylated regions were identified in genes of the pathogenetically significant MET signaling pathway (RNMT, HPN). Differentially methylated regions affecting genes involved in inflammation regulation (IL-23, IL-23R, IL12B, IL12RB1, and FIS1) were detected in SM combined with dysplasia in comparison with SM combined with BCH. The revealed changes in DNA methylation may underlie various "scenarios" of the precancerous process in the bronchial epithelium.

Genetic Features of Tongue Cancer Recurrence in Young Adults.

To identify genetic alterations associated with tongue cancer recurrence in young adults, whole exome sequencing of the primary tumor, recurrence, and whole blood samples from young patients with tongue cancer was performed. A frameshift mutation in the TP53 gene was detected in the primary tumor and recurrence tumor tissue. A mutation in the EPHB6 gene was detected in the recurrence and was absent in the primary tumor. In addition, the primary tumor and recurrence tongue cancer tissue harbored amplification of the 20p13 region containing C20orf96, DEFB125, DEFB126, DEFB127, DEFB128, DEFB129, DEFB132, and ZCCHC3 genes. Thus, genetic alterations have been identified that are associated with tongue cancer recurrence in young adults.

DNA methylome analysis reveals potential alterations contributing to the progression of bronchial hyperplasia.

BACKGROUND: Squamous cell lung cancer (SCLC) arises from bronchial changes: basal cell hyperplasia (BCH), squamous metaplasia (SM), and dysplasia. However, the premalignant process preceding SCLC is not inevitable; it can stop at any of the bronchial lesions. Previously, we hypothesized that combinations of premalignant lesions observed in the small bronchi of SCLC patients can reflect the different "scenarios" of the premalignant process: BCHi-the stoppage at the stage of hyperplasia and BCHSM-the progression of hyperplasia to metaplasia. METHODS AND RESULTS: In this study, using whole-genome bisulfite sequencing we analyzed the DNA methylome of two forms of BCH: isolated BCH (BCHi) and BCH co-occurred with SM (BCHSM) in the small bronchi of SCLC patients. It was shown that BCHi harbored differentially methylated regions (DMRs) affecting genes associated with regulating phosphatase activity. In BCHSM, DMRs were found in genes involved in PI3K-Akt and AMPK signaling pathways. DMRs were also found to affect specific miRNA genes: miR-34a and miR-3648 in BCHi and miR-924 and miR-100 in BCHSM. CONCLUSIONS: Thus, this study demonstrated the significant changes in DNA methylome between the isolated BCH and BCH combined with SM. The identified epigenetic alterations may underlie different "scenarios" of the premalignant process in the bronchial epithelium.

Intravasation as a Key Step in Cancer Metastasis.

Intravasation is a key step in cancer metastasis during which tumor cells penetrate the vessel wall and enter circulation, thereby becoming circulating tumor cells and potential metastatic seeds. Understanding the molecular mechanisms of intravasation is critically important for the development of therapeutic strategies to prevent metastasis. In this article, we review current data on the mechanisms of cancer cell intravasation into the blood and lymphatic vessels. The entry of mature thymocytes into the circulation and of dendritic cells into the regional lymph nodes is considered as example of intravasation under physiologically normal conditions. Intravasation in a pathophysiological state is illustrated by the reverse transendothelial migration of leukocytes into the bloodstream from the sites of inflammation mediated by the sphingosine 1-phosphate interaction with its receptors. Intravasation involves both invasion-dependent and independent mechanisms. In particular, mesenchymal and amoeboid cell invasion, as well as neoangiogenesis and vascular remodeling, are discussed to play a significant role in the entry of tumor cells to the circulation. Special attention is given to the contribution of macrophages to the intravasation via the CSF1/EGF (colony stimulating factor 1/epidermal growth factor) paracrine signaling pathway and the TMEM (tumor microenvironment of metastasis)-mediated mechanisms. Other mechanisms including intravasation of tumor cell clusters surrounded by the vessel wall elements, cooperative intravasation (entry of non-invasive tumor cells to the circulation following invasive tumor cells), and intravasation associated with the vasculogenic mimicry (formation of vascular channels by tumor cells) are also discussed. Novel intravasation-specific mechanisms that have not yet been described in the literature are suggested. The importance of targeted therapeutic strategies to prevent cancer intravasation is emphasized.

Types of Immune-Inflammatory Responses as a Reflection of Cell-Cell Interactions under Conditions of Tissue Regeneration and Tumor Growth.

Inflammatory infiltration of tumor stroma is an integral reflection of reactions that develop in response to any damage to tumor cells including immune responses to antigens or necrosis caused by vascular disorders. In this review, we use the term "immune-inflammatory response" (IIR) that allows us to give an integral assessment of the cellular composition of the tumor microenvironment. Two main types of IIRs are discussed: type 1 and 2 T-helper reactions (Th1 and Th2), as well as their inducers: immunosuppressive responses and reactions mediated by Th22 and Th17 lymphocytes and capable of modifying the main types of IIRs. Cellular and molecular manifestations of each IIR type are analyzed and their general characteristics and roles in tissue regeneration and tumor growth are presented. Since inflammatory responses in a tumor can also be initiated by innate immunity mechanisms, special attention is given to inflammation based on them. We emphasize that processes accompanying tissue regeneration are prototypes of processes underlying cancer progression, and these processes have the same cellular and molecular substrates. We focus on evidence that tumor progression is mainly contributed by processes specific for the second phase of "wound healing" that are based on the Th2-type IIR. We emphasize that the effect of various types of immune and stroma cells on tumor progression is determined by the ability of the cells and their cytokines to promote or prevent the development of Th1- or Th2-type of IIR. Finally, we supposed that the nonspecific influence on the tumor caused by the cytokine context of the Th1- or Th2-type microenvironment should play a decisive role for suppression or stimulation of tumor growth and metastasis.

Effect of small and radical surgical injury on the level of different populations of circulating tumor cells in the blood of breast cancer patients.

Circulating tumor cells (CTCs) constitute a heterogeneous population. Some tumor cells are cancer stem cells (CSCs), while others are in the process of the epithelial-mesenchymal transition (EMT); however, most CTCs are neither stem cells nor in the EMT. This prospective study of 22 patients with nonspecific-type invasive carcinoma of the breast identified different populations of CTCs by flow cytometry in the blood of patients before biopsy, after biopsy and after surgical tumor removal without neoadjuvant chemotherapy. The results showed that minor surgical injury (biopsy) was accompanied by a significant increase in the blood levels of CTCs without signs of the EMT or stemness (Epcam+CD45-CD44-CD24-Ncadh-) and CTCs with signs of stemness and without signs of the EMT (Epcam+CD45-CD44+CD24-Ncadh-). Our results suggest that minor surgical injury to a tumor contributes to the release of CTCs into the bloodstream, including a population of stem cells.

[Health and safety of worker (commentary to the draft ISo 45001)].

The standard ISO 45001 and poor translation of its title in Russian are discussed. The mortality rates from occupational diseases and injuries are compared as well as risk of sudden cardio-vascular death. The priority of occupational health as compared to safety is proven and some ways of amelioration of the system are suggested.

[FEDERAL CLINICAL RECOMMENDATIONS IN DIAGNOSIS, TREATMENT AND PREVENTION OF HEARING LOSS DUE TO NOISE].

Noise induced hearing loss is a slowly developing hearing impairment, caused by occupational exposure to excessive noise levels, constitutes a lesion of the auditory analyzer and clinically manifested as chronic bilateral sensorineural hearing loss. Currently, there is not a treatment that provide a cure of sensorineural hearing loss. Regular, individually tailored treatment should be directed to the pathogenic mechanisms and specific clinical symptoms of hearing loss, as well as the prevention of complications. We recommend using non-drug therapies that can improve blood flow in labyrinth, tissue and cellular metabolism.

Invasive and drug resistant expression profile of different morphological structures of breast tumors.

In order to understand invasive/adhesive and drug resistant properties of intratumor morphological heterogeneity of breast cancer, we compared the expression of genes responsible for the cell adhesion and for the drug resistance between distinct morphological structures of breast tumors. Tubular (hollow-like), alveolar (morula-like), trabecular, solid structures/patterns, and discrete (small) groups of tumor cells were isolated from invasive carcinoma of no special type (n=3) and invasive micropapillary carcinoma (n=1) of the breast using laser microdissection. The gene expression of cadherins, catenins, integrins, ABC transporters, GSTP1, and drug targets was analyzed using qRT-PCR. Expression of catenin genes was identified in almost all structures. In contrast, the expression of cadherin and integrin genes significantly varied depending on the morphological variant. Cadherin expression declined in the row: solid - alveolar and trabecular structures - discrete groups of tumor cells. Expression of integrins declined in the row: solid and alveolar - trabecular structures - discrete groups of tumor cells. For drug resistance genes, trabecular structures more often demonstrated activity of genes coding for ABC transporters compared to other morphological variants. These results indicate that intratumoral morphological heterogeneity in breast cancer correlates with expression profile of adhesion and drug resistance genes reflecting different patterns of invasive growth and responsiveness to chemotherapy.

[THE RISK OF DEVELOPING DISEASE IN FEMALE WORKERS INVOLVED IN MODERN SECTOR EMPLOYMENT AND THE HEALTH OF THEIR CHILDREN].

There was studied the state of health of females in modern sector employment: office workers (group 1) and managers (group 2), and the health of their newborns. There was established the increase of the risk for the development of extragenital disorders in groups 1 and 2 compared with the control. For adreno-genital syndrome OR = 3.83; (CI: 1.15-12.74) in group 1 and OR = 4,50 (CI: 1.24-16.28) Group 2; for respiratory diseases OR = 2.77-2.98 (CI: 1.82- 4.85) in groups 1 and 2; for digestive diseases OR = 2.45 (CI: 1.39-4.32) in group 1 and 2.77 (CI: 1.46-5.26)--in Group 2. The risk of infertility in the history of the group 2 was OR = 2.56 (CI: 1.33-4.92). The frequency of reproductive loss in the history is signficantly higher in group 1 (12,9 ? 1.28), is increasing in group 2 (20.19 ± 2.75) with 9.74 ± 2.12 in 100 females in the control. In complications of pregnancy the frequency of the threat of its interruption increases from 16.99 ± 2.31 cases per 100 females in the group 1 to 21.60 ± 2.82 in Group 2, with 11.79 ± 2.31 in the control (p < 0.01). In newborn of these female workers there was revealed an increase in the frequency of the congenital defects of development (CDD) of the circulatory system in group 1 by 1.5 times in the group 2-2.5 times if compared to the control (16.38 ± 1.41 and 24.88 ± 2.96, with 9.74 ± 2.12 cases per 100 infants (p < 0.01; p < 0.001), respectively, the frequency of congenital malformations of the urinary system in groups 1 and 2 (10.00 ± 1.14 and 10.80 ± 2.13), was significantly higher than in control--3.08 ± 1.24 per 100 births (p < 0.01); CDD of genitalia was significantly higher in group 1 (11.01 ± 1.19) with 5.13 ± 1.58 per 100 infants in the control (p < 0.01). Thus, disorders of reproductive health in females involved in modern sector employment depend on the degree of occupational hazard factors; can lead to congenital disease in their newborn, which requires the development of preventive measures.

[Bases of information hygiene: concepts and problems of innovations].

The rising flow of information affects the health and affords ground for strengthening measures of hygienists. There is considered the place of information hygiene in preventive medicine. There are discussed physical and hygienic aspects of measurement and evaluation of information, there are considered some of their negative consequences. International and local regulations and guidance documents in this area are presented. In the light of the strategy of innovative development of Russia there are considered hygienic aspects of innovation and the improvement of hygienic standards.

[Criteria for the assessment of the noise-induced occupational hearing loss: international and national standards].

The objective of the present paper was to review international and national standards for the criteria of assessment of the noise-induced occupational loss of hearing. The importance of healthy hearing for the occupational safety is emphasized which implies the necessity of the more rigorous criteria for hearing conservation in the workers engaged in the noisy technological environment compared with those for the general population. A rationale for the development of the standard program of hearing conservation in the workers engaged in the noisy technological environment is proposed including hygienic norms and rules or national state standards.

[Information hygiene and regulation of information for vulnerable groups of the population].

Development of information society engenders the problem of hygienic regulation of information load for the population, first of all for vulnerable groups. There are presented international and Russian normative legal documents and experience in this area, there are described the negative effects of information (such as stress, depression, suicidal ideations). There are considered social-psychological characteristics of vulnerable groups that requires their best protection from loads of information, doing harm, particularly in terms of reproductive health, family relationships, children, etc. There was noted the desirability of improvement of sanitary, legislation on the regulation of the information load on the population, especially in vulnerable groups, in terms of optimization of parameters of the signal-carriers on volume, brightness and the adequacy of the volume and content of information in radio and television broadcasting, in an urban environment and at the plant to preserve the health and well-being of the population.

[Occupational hearing loss--problem of health and safety].

On the basis of the literature review the critical analysis of the recommendations (the letter of Ministry of Health of Russia from 6/11/2012 N 14-1/10/2-3508) on occupation noise-induced hearing loss (HL) assessment is presented. Need of more strict criteria of HL assessment for workers, than for the general population according to ICF (WHO, 2001), in order to avoid growth of accidents and injury rate is proved. The illegitimacy of a deduction of statistical presbiacusia values from individual audiograms as human rights violation is stressed. Some terminological defects are noted. It is necessary to cancel recommendations and to develop the sanitary norms or state standard with the program of hearing conservation at work.

[Information as physical factor: problems of measurement, hygienic assessment and IT-automation].

The increasing flow of information, speeding up the progress of society, can impact the health that puts the task of its hygienic reglamentation. The physical aspects of information, parameters and units of quantities, aspects of measurement and evaluation with account of information quantity and quality as well as criteria of its permissible and optimal levels are considered. The results of measurements of quantity of text information produced per year on computer in 17 occupations of 10 economic sectors are presented. The principle of IT-automation of operator's work and of dynamic monitoring is proposed. On the basis of research performed the glossary of terms and guide on the problem with computer support are elaborated for the accumulation of experience and clarification of prospects.

EpCAM-CD24+ circulating cells associated with poor prognosis in breast cancer patients.

Following the discovery of circulating tumor cells (CTCs) in the peripheral blood of cancer patients, CTCs were initially postulated to hold promise as a valuable prognostic tool through liquid biopsy. However, a decade and a half of accumulated data have revealed significant complexities in the investigation of CTCs. A challenging aspect lies in the reduced expression or complete loss of key epithelial markers during the epithelial-mesenchymal transition (EMT). This likely hampers the identification of a pathogenetically significant subset of CTCs. Nevertheless, there is a growing body of evidence regarding the prognostic value of such molecules as CD24 expressing in the primary breast tumor. Herewith, the exact relevance of CD24 expression on CTCs remains unclear. We used two epithelial markers (EpCAM and cytokeratin 7/8) to assess the count of CTCs in 57 breast cancer patients, both with (M0mts) and without metastasis (M0) during the follow-up period, as well as in M1 breast cancer patients. However, the investigation of these epithelial markers proved ineffective in identifying cell population expressing different combinations of EpCAM and cytokeratin 7/8 with prognostic significance for breast cancer metastases. Surprisingly, we found CD24+ circulating cells (CCs) in peripheral blood of breast cancer patients which have no epithelial markers (EpCAM and cytokeratin 7/8) but was strongly associated with distant metastasis. Namely, the count of CD45-EpCAM-CK7/8-CD24+ N-cadherin-CCs was elevated in both groups of patients, those with existing metastasis and those who developed metastases during the follow-up period. Simultaneously, an elevation in these cell counts beyond the established threshold of 218.3 cells per 1 mL of blood in patients prior to any treatment predicted a 12-fold risk of metastases, along with a threefold decrease in distant metastasis-free survival over a 90-month follow-up period. The origin of CD45-EpCAM-CK7/8-CD24+ N-cadherin-CCs remains unclear. In our opinion their existence can be explained by two most probable hypotheses. These cells could exhibit a terminal EMT phenotype, or it might be immature cells originating from the bone marrow. Nonetheless, if this hypothesis holds true, it's worth noting that the mentioned CCs do not align with any of the recognized stages of monocyte or neutrophil maturation, primarily due to the presence of CD45 expression in the myeloid cells. The results suggest the presence in the peripheral blood of patients with metastasis (both during the follow-up period and prior to inclusion in the study) of a cell population with a currently unspecified origin, possibly arising from both myeloid and tumor sources, as confirmed by the presence of aneuploidy.

Intratumor heterogeneity: nature and biological significance.

Intratumor heterogeneity inherent in the majority of human cancers is a major obstacle for a highly efficient diagnosis and successful prognosis and treatment of these diseases. Being a result of clonal diversity within the same tumor, intratumor heterogeneity can be manifested in variability of genetic and epigenetic status, gene and protein expression, morphological structure, and other features of the tumor. It is most likely that the appearance of this diversity is a source for the adaptation of the tumor to changes in microenvironmental conditions and/or a tool for changing its malignant potential. In any case, both processes result in the appearance of cell clones with different undetermined sets of hallmarks. In this review, we describe the heterogeneity of molecular disorders in various human tumors and consider modern viewpoints of its development including genetic and non-genetic factors of heterogeneity origin and the role of cancer stem cells and clonal evolution. We also systematize data on the contribution of tumor diversity to progression of various tumors and the efficiency of their treatment. The main problems are indicated in the diagnosis and therapy of malignant tumors caused by intratumor heterogeneity and possible pathways for their solution. Moreover, we also suggest the key goals whose achievement promises to minimize the problem of intratumor heterogeneity and to identify new prognostic, predictive, and target markers for adequate and effective treatment of cancer.

[Correlation of metastasis-free survival in patients with breast cancer and changes in the direction of expression of multidrug resistance genes during neoadjuvant chemotherapy].

Previously, we showed the association of neoadjuvant chemotherapy (NAC) response with changing the expression vector (increase or decrease) of multidrug resistance genes (MDR) in breast tumors during chemotherapy. The aim of the present study was to evaluate the relation between changes in the expression vector of MDR genes and distant metastasis-free survival. Patients (n = 120) with breast cancer (T1-4N0-3M0) treated by 2-4 cycles of NAC (CAX, FAC, and taxane regimes) and 4 cycles of adjuvant chemotherapy (FAC) were included. TaqMan-based quantitative reverse transcriptase PCR (qRT-PCR) was used to estimate the expression of the following MDR genes: ABCB1, ABCC1, ABCC2, ABCC5, ABCG1, ABCG2, GSTP1, and MVP--in biopsies before NAC and in tumor samples after chemotherapy. Comparing the corresponding expression levels allowed us to identify the vector of expression change during NAC. The results showed that 5-year distant metastasis-free survival was 73-78% in patients with a decrease in ABCB1, ABCC2, and ABCG1 expression. The up-regulation of these genes during NAC was related to a significant decrease (up to 50-55%) in metastasis-free survival (Kaplan-Meier analysis: log-rank p value = 0.006-0.03). The association of changing the expression vector of MDR genes with metastasis-free survival did not depend on tumor size, lymph node involvement, histological form, receptor status, molecular subtype, and others clinicopathological parameters of breast cancer. The obtained data suggest that changing the expression vector of MDR genes in breast tumors during NAC may be used as a new potential prognostic marker of breast cancer. An increase in tumor expression of ABCB1, ABCC2, and ABCG1 during chemotherapy is a factor of poor prognosis, whereas down-regulation of these genes--a favorable prognostic marker.

[Reformation of occupational health in France: state of the problem and new law].

The state of the problem and new law of France No 2011-867 on occupational health organization and two decrees are described. These introduce services of health at work mainly with prophylactic tasks and whose functions are performed by interdisciplinary teams including doctors, assistants and specialists on occupational risk prevention. The documents are based on ILO Convention No 161 "Occupational health services".

[The real-world effectiveness of personal protective equipment and additional risks for workers' health].

The effectiveness of personal protective equipment (PPE) of hearing, respiratory organs and hands is considered. It is shown that real effect of PPE is twice lower than declared by supplier; this presumes some derating system. The aspects of discomfort and additional risks are analyzed. The hygienic and physiologic evaluation of PPE is required along with elaboration of an official document (OSH standard or sanitary regulation) on selection, personal fit, organization of use and individual training of workers and their motivation.