Host genetic variants associated with COVID-19 reconsidered in a Slovak cohort.

We present the results of an association study involving hospitalized coronavirus disease 2019 (COVID-19) patients with a clinical background during the 3rd pandemic wave of COVID-19 in Slovakia. Seventeen single nucleotide variants (SNVs) in the eleven most relevant genes, according to the COVID-19 Host Genetics Initiative, were investigated. Our study confirms the validity of the influence of LZTFL1 and 2'-5'-oligoadenylate synthetase (OAS)1/OAS3 genetic variants on the severity of COVID-19. For two LZTFL1 SNVs in complete linkage disequilibrium, rs17713054 and rs73064425, the odds ratios of baseline allelic associations and logistic regressions (LR) adjusted for age and sex ranged in the four tested designs from 2.04 to 2.41 and from 2.05 to 3.98, respectively. The OAS1/OAS3 haplotype 'gttg' carrying a functional allele G of splice-acceptor variant rs10774671 manifested its protective function in the Delta pandemic wave. Significant baseline allelic associations of two DPP9 variants in all tested designs and two IFNAR2 variants in the Omicron pandemic wave were not confirmed by adjusted LR. Nevertheless, adjusted LR showed significant associations of NOTCH4 rs3131294 and TYK2 rs2304256 variants with severity of COVID-19. Hospitalized patients' reported comorbidities were not correlated with genetic variants, except for obesity, smoking (IFNAR2), and hypertension (NOTCH4). The results of our study suggest that host genetic variations have an impact on the severity and duration of acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Considering the differences in allelic associations between pandemic waves, they support the hypothesis that every new SARS-CoV-2 variant may modify the host immune response by reconfiguring involved pathways.

The potential use of miRNAs in the diagnosis and prediction of metastatic lung carcinoma.

Lung carcinoma is the "top killer" of all malignancies in the world. Early diagnosis of lung carcinoma significantly improves patient survival. Screening with biomarkers from peripheral blood could detect more patients at an early stage of the disease. MicroRNAs (miRNAs) could be a possible biomarker. These are 21-23 nucleotide long single-stranded RNA molecules playing an important role in the post-transcriptional regulation of gene activity. Individual miRNAs have the potential to regulate genes responsible for cell proliferation, differentiation, apoptosis, regulate cell cycle in cooperation with pro-oncogenes and tumor suppressor genes. In our study, we determined miRNA expression levels in individual samples of lung carcinoma patients and in a healthy control group. We used the reverse transcription method followed by qRT-PCR. The expression levels of the investigated miRNAs were evaluated in the QIAGEN GeneGlobe Data center software. We demonstrated the significance of miR-126 and let-7g as biomarkers of lung carcinoma in all clinical stages studied. We also observed significantly increased expression of miR-143 and miR-145 at the distant metastasis stage, and significantly decreased expression of miR-133a in the N2 disease group of lung carcinoma patients (N2 disease represents disease with metastases in the ipsilateral mediastinal and/or subcarinal lymph nodes or node). The investigated miRNAs showed no clear potential for detecting potentially resectable (N0-N1), locally advanced (N2) and distant organ metastatic (M1) lung carcinoma.

Risk genetic polymorphism and haplotype associated with papillary thyroid cancer and their relation to associated diseases in Slovak population.

BACKGROUND: Risk for developing papillary thyroid carcinoma (PTC), the most common endocrine malignancy, is thought to be mediated by lifestyle, environmental exposures and genetic factors. Recent progress in the genome-wide association studies of thyroid cancer leads to the identification of several genetic variants conferring risk to this malignancy across different ethnicities. METHODS AND RESULTS: We set out to elucidate the impact of selected single nucleotide polymorphisms (SNPs) on papillary thyroid carcinoma risk and to evaluate the interactions of these genetic variants with associated diseases for the first time in the Slovak population. Six SNPs (rs966423, rs2439302, rs965513, rs116909374, rs1537424 and rs944289) were genotyped in 86 patients with PTC and 99 healthy control subjects. The association analysis and multivariable modelling of PTC risk by the genetic factors, supplemented with a rigorous statistical validation, were performed. One of the six SNPs rs966423 (DIRC3, OR=1.51, p=0.03) was significantly associated with PTC. Next two SNPs rs965513 (PTCSC2, OR=1.34) and rs116909374 (MBIP, OR=0.44) showed a suggestive association. Haplotype TTC (SNPs located on chromosome 14q13) showed a suggestive association with PTC (p=0.07, OR=1.55). In the PTC group, significant associations were observed between rs966423 (DIRC3) and ischemic heart diseases (p=0.009), rs965513 (PTCSC2) and diabetes mellitus (p=0.04) and haplotype 14q13 and musculoskeletal diseases. Next three associations rs966423 (DIRC3) and arterial hypertension; rs116909374 (MBIP) and other benign diseases; rs1537424 (MBIP) and disorder lipid metabolism, rs965513 (PTCSC2) and anti-Tg (thyroglobulin antibody) showed suggestive associations. CONCLUSION: These results indicate that germline variants not only predispose to PTC, but may also be related to other risk factors, including associated diseases. However, these associations were only moderate, and further multi-ethnic studies are required to evaluate the usefulness of these germline variants in the clinical stratification of PTC patients (Tab. 8, Ref. 37).

Overexpression of the miR-143/145 and reduced expression of the let-7 and miR-126 for early lung cancer diagnosis.

INTRODUCTION: Lung cancer is the leading cause of cancer-related deaths worldwide. For this reason, huge efforts are being invested in discovering suitable blood biomarkers that would allow early diagnosis and treatment. One of the possible promising candidates for this role are microRNA molecules (miRNAs). The aim of the study was to identify individual blood miRNAs that could be used as potential biomarkers for early diagnosis of lung cancer. METHODS: This prospective study analyzed blood samples of 60 patients with early-stage lung cancer, and blood samples of 60 healthy individuals. All study patients with lung cancer had undergone radical pulmonary resection at the University Hospital Ostrava within the study period (2015-2017). Definitive diagnosis of lung cancer was confirmed by histopathology examination of the resected pulmonary specimen. We investigated relative expressions in selected 13 blood miRNAs; the examined miRNAs were miR-126, miR-155, miR-221, miR-21, miR-143, miR-145, miR-133a, let-7a, miR-146a, miR-31, miR-182, let-7g and miR-19b. RESULTS: The outcome of this study showed that the levels of the majority of the tested circulating miRNA in lung cancer patients are significantly altered. The most significant serum miRNA biomarkers for the early detection of lung cancer are as follows: miR-143, let-7g, miR-126, let-7a, and miR-145 (miR-143 and miR-145 have oncogene functions, while miR-126, let-7g and let-7a have suppressor functions). CONCLUSIONS: We have demonstrated the excellent diagnostic value of several miRNAs (miR-126, miR-143, miR-145, let-7a and let7g). These have an estimated sensitivity and specificity of 75-85% and 0.90-0.93 AUC. However, these individual miRNA biomarkers require further validation in larger prospective cohorts.

Effect of Cold Atmospheric Plasma on Epigenetic Changes, DNA Damage, and Possibilities for Its Use in Synergistic Cancer Therapy.

Cold atmospheric plasma has great potential for use in modern medicine. It has been used in the clinical treatment of skin diseases and chronic wounds, and in laboratory settings it has shown effects on selective decrease in tumour-cell viability, reduced tumour mass in animal models and stem-cell proliferation. Many researchers are currently focusing on its application to internal structures and the use of plasma-activated liquids in tolerated and effective human treatment. There has also been analysis of plasma's beneficial synergy with standard pharmaceuticals to enhance their effect. Cold atmospheric plasma triggers various responses in tumour cells, and this can result in epigenetic changes in both DNA methylation levels and histone modification. The expression and activity of non-coding RNAs with their many important cell regulatory functions can also be altered by cold atmospheric plasma action. Finally, there is ongoing debate whether plasma-produced radicals can directly affect DNA damage in the nucleus or only initiate apoptosis or other forms of cell death. This article therefore summarises accepted knowledge of cold atmospheric plasma's influence on epigenetic changes, the expression and activity of non-coding RNAs, and DNA damage and its effect in synergistic treatment with routinely used pharmaceuticals.

Metabolomic profiling of blood plasma of patients with lung cancer and malignant tumors with metastasis in the lungs showed similar features and promising statistical discrimination against controls.

Targeting metabolomic pathways is a promising strategy for cancer treatment. Alterations in the metabolomic state have also an epigenetic impact, making the metabolomic studies even more interesting. We explored metabolomic changes in the blood plasma of patients with primary and secondary lung cancer and tried to explore their origin. We also applied a discrimination algorithm to the data. In the study, blood samples from 132 patients with primary lung cancer, 47 with secondary lung cancer, and 77 subjectively healthy subjects without any cancer history were used. The samples were measured by NMR spectroscopy. PCA and PLS-DA analyses did not distinguish between patients with primary and secondary lung tumors. Accordingly, no significantly changed levels of plasmatic metabolites were found between these groups. When comparing with healthy controls, significantly increased glucose, citrate, acetate, 3-hydroxybutyrate, and creatinine balanced with decreased pyruvate, lactate, alanine, tyrosine, and tryptophan were found as a common feature of both groups. Metabolomic analysis of blood plasma showed considerable proximity of patients with primary and secondary lung cancer. The changes observed can be partially explained as cancer-derived and also as changes showing ischemic nature. Random Forrest discrimination based on the relative concentration of metabolites in blood plasma performed very promising with AUC of 0.95 against controls; however noticeable parts of differencing metabolites are overlapping with those observed after ischemic injury in other studies.

Common gene haplotypes of gelatinases and their tissue inhibitors in abdominal aortic aneurysm.

Abdominal aortic aneurysm (AAA) involves complex dynamic remodeling processes in the aortic wall. Gelatinases (MMP2 and MMP9) and their respective tissue inhibitors (TIMP1 and TIMP2) play a crucial role during extracellular matrix (ECM) turnover in aortic tissue. In this study we characterized associations between the haplotypes of genes encoding gelatinase/inhibitor pairs and pathways involved in AAA, a total of 100 AAA patients and 192 controls were enrolled. For males, a significant decrease in the distribution of the minor G allele of the TIMP2 rs8082025 was observed in AAA patients (p = 0.01, 23.1% controls vs. 13.1% AAA). In addition, in males, the major TIMP2 GA haplotype was associated with AAA (86.9% AAA vs. 76.9% control; p = 0.009, OR = 1.997), whereas the TIMP2 GG haplotype (7.7% AAA vs. 13.9% control) was associated with protection against AAA (p = 0.046, OR = 0.518). The minor GAGC MMP9 haplotype was related to AAA for all study subjects as well as the males only subset (p = 0.011, OR = 2.202 and p = 0.025, OR = 2.156, respectively). Small differences in the distribution of gene haplotypes could be associated with different levels of gene expression and in turn influence gelatinases activity in AAA.

The Role of Interleukin-6 Polymorphism (rs1800795) in Prostate Cancer Development and Progression.

BACKGROUND/AIM: Interleukin-6 is an important modulator of inflammation, which is one of the factors involved in prostate cancer. The aim of the study was to evaluate the possible association of the IL-6 -174 polymorphism (rs1800795) with the risk of prostate cancer development and progression. MATERIALS AND METHODS: The study population consisted of 446 prostate cancer patients, 377 benign prostatic hyperplasia (BHP) patients and 276 healthy men. Genotyping was performed by PCR-RFLP analysis. IL-6 plasma levels were measured by the ELISA method. RESULTS: The GC genotype (OR=0.61, p=0.005) and C allele (OR=0.8, p=0.04) of the IL-6 -174 polymorphism were significantly associated with prostate cancer. No genotype was associated with BHP. IL-6 plasma levels were significantly increased in prostate cancer patients compared to both healthy men (p=0.02) and BHP patients (p=0.008). No significant differences were observed in IL-6 plasma levels in connection with IL-6 -174 genotypes. CONCLUSION: The IL-6 -174 polymorphism was significantly associated with prostate cancer in Slovak patients.

Low Variability and Stable Frequency of Common Haplotypes of the TP53 Gene Region in Colorectal Cancer Patients in a Slovak Population.

AIM: To test the association between common TP53 haplotypes and colorectal cancer (CRC) development. PATIENTS AND METHODS: A total of 277 CRC patients and 167 healthy volunteers were included in the study. Common TP53 haplotypes were estimated from eight single-nucleotide polymorphisms (SNPs) (rs1614984, rs77697176, rs12947788, rs1800372, rs2909430, rs1042522, rs17878362 and rs11652704). Stepwise haplotype trend regression showed the haplotype-regressor cccgaRDa as a possible predictive marker. RESULTS: The rare haplotype cccgaRDa was identified in 10 CRC cases and 3 controls. Although it is approximately twice as common in CRC (odds ratio (OR)=2.068; 95% confidence interval (CI)=0.471-9.069), the cccgaRDa haplotype frequency is low in the studied groups. Results of our study suggest that the common TP53 variability is relatively low (only 3 haplotypes occurred above 10%). CONCLUSION: The haplotype background of TP53 gene is relative stable and despite low haplotype-regressor cccgaRDa frequency it shows to be a possible predictive parameter for CRC development.

DPYD genotype and haplotype analysis and colorectal cancer susceptibility in a case-control study from Slovakia.

Dihydropyrimidine dehydrogenase (DPD) acts as the first-step enzyme catabolizing pyrimidines in vivo. DPYD gene mutations interfere with the breakdown of uracil and thymine. Genetic variations of DPYD can cause an enzyme deficiency state, which results in severe toxicity or other adverse side effects such as DNA damage or RNA damage caused by imbalance of the nucleotide pool. Our case-control study investigates the possible association between seven DPYD gene polymophisms (rs1801267, rs72547602, rs1801160, rs3918290, rs1801159, rs1801158, rs1801265) and risk of colorectal cancer (CRC). The association analysis for DPD was performed on 273 CRC patients and 187 healthy controls. There is significant allele association of SNP rs1801160 with colorectal cancer (p = 0.003, OR = 3.264, 95% CI = 1.425-7.475) in present analysis. Haplotype analysis of four DPYD polymorphisms showed significant difference in the distribution "IISt" haplotype between cases and controls. In comparison to the most common haplotype (VISt), the "IISt" haplotype was associated with increased risk for CRC (p = 0.038, OR = 2.733, 95% CI = 1.019-7.326). The present study suggests that the SNP rs1801160 and the "IISt" haplotype in the DPYD gene may also have a role in colorectal cancer risk.

Effect of CYP17 and PSA gene polymorphisms on prostate cancer risk and circulating PSA levels in the Slovak population.

Cytochrome P-450c17α (CYP17) and prostate-specific antigen (PSA) genes, which are involved in the androgen metabolism cascade, have been studied as possible candidates for genetic influences on prostate cancer development. Contradictory results prompted us to evaluate the frequencies of polymorphisms in the CYP17 and PSA genes as well as the association between these genetic variants and serum PSA levels in prostate cancer patients and men routinely screened for prostate cancer with PSA in the Slovak male population. The CYP17 and PSA polymorphisms were determined by the PCR-RFLP analysis in 197 Caucasian prostate cancer patients and 256 Caucasian controls. We did not find any association between the CYP17 and PSA genotypes and prostate cancer risk overall, or by grade. Also the total serum PSA levels in the cases with the AG or AA genotype were not significantly higher than in the men with the GG genotype (P > 0.05). Our study did not provide support for the hypothesized relationship between CYP17 and PSA gene polymorphisms and prostate cancer in the Slovak male population.

Evaluating chromosomal damage in workers exposed to hexavalent chromium and the modulating role of polymorphisms of DNA repair genes.

PURPOSE: Welders have been chronically exposed to hexavalent chromium with potential consequences on chromosomal integrity. Our study is focused on the extent of any such chromosomal aberrations with respect to chromium levels in the blood of welders as well as on the tentative modulating role of polymorphisms in DNA repair genes XPD Lys751Gln, XPG Asn114His, XPC Lys939Gln, hOGG1 Ser326Cys and XRCC1 Arg399Gln on chromosomal damage. METHODS: The study was conducted on 144 individuals consisting of 73 welders exposed to chromium for 10.2 ± 1.67 years and 71 control individuals without known exposures. Chromosomal aberrations, their chromatid-type and chromosome-type aberrations were detected by conventional cytogenetic analysis. XPD, XPG, XPC, hOGG1 and XRCC1 gene polymorphisms were assayed for by Taqman SNP genotyping assay ("Assay-by-Demand") using Real-Time allelic discrimination on AB 7500 equipment. Chromium concentration in the blood was determined by atomic absorption spectrophotometry. RESULTS: The level of chromium in the blood of welders ranged between 0.032 and 0.182 µmol l(-1) and was significantly higher than that in controls (0.07 ± 0.04 µmol l(-1) vs. 0.03 ± 0.007 µmol l(-1)). Parameters of chromosomal damage were similar in both the exposed and the control individuals (1.89% vs. 1.70% for total chromosomal aberrations, 0.97% vs. 0.88% for chromosome-type and 0.92% vs. 0.80% for chromatid-type, respectively). Chromatid-type of aberrations positively correlated with the level of chromium in the blood (r = 0.28; P = 0.02). Significantly higher total chromosomal aberrations were detected in individuals with homozygous variant polymorphism in XRCC1 Arg399Gln gene as compared to those with heterozygous and homozygous wild-type genotypes (2.20, 1.89 and 1.48%, respectively; P = 0.01). A similar tendency was found for chromatid-type aberrations (1.30% for homozygous variant genotype bearers, 0.94% for those with heterozygous genotype and 0.75% for carriers of homozygous wild-type genotype, respectively; P = 0.04). CONCLUSIONS: Although no apparent increase in chromosomal damage was recorded in chromium-exposed welders in comparison with controls, genetic make-up in DNA repair genes may increase susceptibility toward adverse effect of chromium.

Antitussive activity of an extracellular Rhodella grisea proteoglycan on the mechanically induced cough reflex.

A mucilagineous extracellular proteoglycan (EPG) composed of xylose and its 3-O-and 4-O-methyl-derivates (55%), glucuronic acids (17%), rhamnose (14%), galactose (8%), glucose (4%) and minor amounts of other sugars (∼2%) has been isolated from culture medium of Rhodella grisea. A white fluffy algal biopolymer of molecular mass over 8.1×105 contained protein (13%), methoxyl (6%), acetyl and succinyl groups. EPG was tested in vivo on mechanically induced cough in non-anaesthetized cats as a test system. The biopolymer showed a cough suppressing effect on laryngopharyngeal type of cough while the cough from tracheobronchial mucous area was slightly or not affected. Further, the intensity of maximal cough efforts from laryngopharyngeal and tracheobronchial parts in expirium and inspirium were influenced slightly only indicating that the expectoration effect was not suppressed by biopolymer application.

Human lung cancer and hexavalent chromium exposure.

OBJECTIVE: Workers chronically exposed to hexavalent chromium have higher incidence of lung cancer. Our study investigates incidence of lung cancer types, age at onset of the disease and survival time among chromium exposed workers (smelters, tapers, crane operators) in comparison to non-exposed persons. METHODS: 64 chromium exposed workers and 104 male controls with diagnosed lung cancer were analysed. The average exposure time among workers was 16.71 +/- 10.02 (S.D.) years (range 1- 41 years). RESULTS: Chromium exposure significantly decreases the age at the onset of the disease by 3.51 years (62.20 +/- 9.08 years in exposed group and 65.71 +/- 10.50 years in control; P=0.018). Small cell lung carcinoma (SCLC) forms 25.0 % of all cases in chromium exposed workers and 16.34% in non exposed individuals. No correlation was found between the age at the diseases onset and time of exposure. The mean survival time in exposed group was 9.03 +/- 12.73 month, in control 12.14 +/- 21.94 month, but this difference was not significant (P=0.473). CONCLUSION: Occupational exposure to chromium was identified as an important risk factor of lung cancer, decreasing the age at the diseases onset. Higher percentage of SCLC was found in chromium exposed individuals.

Polymorphisms of glutathione-S-transferase M1, T1, P1 and the risk of prostate cancer: a case-control study.

BACKGROUND: It has been suggested that polymorphisms in glutathione-S-transferases (GST) could predispose to prostate cancer through a heritable deficiency in detoxification pathways for environmental carcinogens. Yet, studies linking GST polymorphism and prostate cancer have so far failed to unambiguously establish this relation in patients. A retrospective study on healthy, unrelated subjects was conducted in order to estimate the population GST genotype frequencies in the Slovak population of men and compare our results with already published data (GSEC project-Genetic Susceptibility to Environmental Carcinogens). A further aim of the study was to evaluate polymorphisms in GST also in patients with prostate cancer in order to compare the evaluated proportions with those found in the control subjects. METHODS: We determined the GST genotypes in 228 healthy, unrelated subjects who attended regular prostate cancer screening between May 2005 and June 2007 and in 129 histologically verified prostate cancer patients. Analysis for the GST gene polymorphisms was performed by PCR and PCR-RFLP. RESULTS: We found that the GST frequencies are not significantly different from those estimated in a European multicentre study or from the results published by another group in Slovakia. Our results suggest that Val/Val genotype of GSTP1 gene could modulate the risk of prostate cancer, even if this association did not reach statistical significance. We did not observe significantly different crude rates of the GSTM1 and GSTT1 null genotypes in the men diagnosed with prostate cancer and those in the control group. CONCLUSION: Understanding the contribution of GST gene polymorphisms and their interactions with other relevant factors may improve screening diagnostic assays for prostate cancer. We therefore discuss issues of study feasibility, study design, and statistical power, which should be taken into account in planning further trials.

In vitro assays for the evaluation of drug resistance in tumor cells.

Oncologic diseases are among leading cause of mortality in developed countries. Despite significant progress, the use of standard cytotoxic chemotherapy has reached a therapeutical plateau. Currently, the process of selecting chemotherapy represents a trial and error method neglecting biological individuality of tumor and its bearer. The improvement of treatment results is expected from ex vivo drug sensitivity testing which may allow to choose the most effective drug for individual patient and to exclude agents to which the tumor cells exert resistance. New techniques and rapidly increasing knowledge about the molecular basis of malignant diseases provide important opportunities for the future of chemotherapy. This paper reviews current methods used to test the resistance of tumor cells to a panel of anticancer agents in vitro. In addition, we focused on the in vitro MTT assay which represents one of major technique for testing of tumor cell resistance to anticancer agents.

Comparison of chromosomal aberrations frequency and polymorphism of GSTs genes in workers occupationally exposed to cytostatics or anaesthetics.

Authors compared the incidence of chromosomal aberrations (CAs) of workers occupationally exposed to cytostatics (group EXP1) or anaesthetics (group EXP2) in relationship to polymorphism of GSTM1, GSTP1 and GSTT1 genes. The cytogenetic analysis for chromosomal aberrations frequency and for polymorphisms of genes the PCR and PCR-RFLP method were used. Statistically higher frequency of total CAs was detected in both exposed groups: group EXP1 1.90±1.34%; Mann-Whitney U-test, p=0.001; group EXP2 2.53±1.46%, p=0.0008) as compared to control (1.26±0.93%). In group EXP2 was detected statistically higher frequency of aberrations CSA-type as compared to CTA-type. In xenobiotic metabolizing genes for GST higher frequency of total CAs and constituent types chromatid-type aberrations (CTAs) and chromosome-type aberrations (CSAs) of genes GSTM1 and GSTT1 with null genotype was detected. Statistically significant difference was detected only in CSA-type of aberrations in GSTT1 gene. In gene GSTP1 was not detected any difference in frequency of aberrations in presence of the variant allele. Presented results point out importance of individual susceptibility in evaluation of genotoxic agents of anaesthetics or cytostatics.

Chromosomal damage and polymorphisms of DNA repair genes XRCC1 and XRCC3 in workers exposed to chromium.

OBJECTIVES: Welders, chronically exposed to hexavalent chromium, may exhibit disturbances in chromosomal integrity. Our study investigates chromosomal damage related to chromium exposure, considering the role of polymorphisms in relevant DNA repair genes. METHODS: 39 male welders exposed to chromium for 10.2+/-1.67 years and 31 male controls were assayed for structural chromosomal aberrations (conventional cytogenetic analysis). DNA repair gene polymorphisms were determined by Real-Time allelic discrimination assay. RESULTS: Total chromosomal aberrations were moderately, but statistically not significant, higher in exposed individuals (1.96%) than in controls (1.55%). Chromosomal type breaks were almost two-fold higher in exposed than in control individuals. The highest frequency of total chromosomal aberrations was recorded in individuals with homozygous variant Gln/Gln cariers (2.14%) in XRCC1* Arg-399Gln and the lowest in those with the wild-type Arg/Arg cariers (1.33%). Polymorphisms in XRCC3 gene did not modulate the frequencies of CAs, CTAs and CSAs. CONCLUSIONS: Understanding the effects of chromium on chromosomal integrity in relation to individual susceptibility may be a basis for preventive measures in working process.