Is a Proactive Approach to Controlling Legionella in the Environment Justified?

RESEARCH BACKGROUND: Legionella are Gram-negative bacteria that are ubiquitous in the natural environment. Contaminated water in man-made water systems is a potential source of transmission of legionnaires' disease. The aim of this study is to explore the prevalence of Legionella pneumophila in the drinking water distribution system (DWDS) of Primorje-Gorski Kotar (PGK) County, Croatia, for the period 2013-2019, coupled with the incidence of legionnaires' disease. A number of L. pneumophila-positive samples (>100 CFU/L), serogroup distribution and the degree of contamination of specific facilities (health and aged care, tourism, and sports) were assessed. Based on the obtained results, the reasoning for the implementation of a mandatory Legionella environmental surveillance program was assessed. EXPERIMENTAL APPROACH: Sample testing for Legionella was carried out according to the standard method for enumeration of this bacterium. A heterotrophic plate count (HPC) and Pseudomonas aeruginosa number were analysed along with the basic physicochemical indicators of drinking water quality. The research period was divided into two parts, namely, the 2013-2018 period (before implementation of the prevention program, after the outbreak of legionnaires' disease), and the year 2019 (proactive approach, no disease cases recorded). RESULTS AND CONCLUSION: During the 7-year observation period in PGK County, an increase in the number of samples tested for Legionella was found. An increase in Legionella-positive samples (particularly pronounced during the warmer part of the year) was recorded, along with a growing trend in the number of reported legionnaires' disease cases. In addition to hot water systems, the risk of Legionella colonisation also applies to cold water systems. Health and aged care facilities appear to be at highest risk. In addition to the higher proportion of positive samples and a higher degree of microbiological load in these facilities, the highest proportion of L. pneumophila SGs 2-14 was identified. Due to the diagnostic limitations of the applied tests, the number of legionnaires' disease cases is underdiagnosed. NOVELTY AND SCIENTIFIC CONTRIBUTION: The introduction of a mandatory preventive approach to monitoring Legionella in DWDS water samples, along with the definition of national criteria for the interpretation of the results will create the preconditions for diagnosis and adequate treatment of larger numbers of legionnaires' disease cases.

Impact of antibiotic consumption on the carriage of antibiotic-resistant bacteria by school children.

BACKGROUND: Antibiotic consumption in the paediatric population is one of the key drivers of the emergence and spread of antimicrobial resistance, which is a serious global threat to public health and clinical medicine. The aims of this study were to investigate systemic antibiotic consumption in school children and to assess the associations among antibiotic consumption, carriage rate and resistance of respiratory pathogens residing in the upper respiratory tract mucosa. METHODS: In this prospective study, throat and nasopharyngeal swabs from 450 school children, 6-15 years of age (225 healthy children and 225 patients who were ambulatory treated for upper respiratory tract infection), were processed in 2014 in Rijeka, Croatia, and clinical data were obtained via a questionnaire. RESULTS: In total, 17% of the children had consumed an antibiotic in the previous 6 months, including 7% of the healthy children and 27% of the acutely ill patients. The most commonly prescribed antibiotics were amoxicillin (26%), amoxicillin with clavulanic acid (26%) and macrolides (18%). Respiratory pathogens were more frequently isolated from children who had consumed an antibiotic in the previous 6 months [odds ratio (OR) 3.67, P < 0.001]. Antibiotic-resistant bacteria were also more frequent in children who had been exposed to antibiotics (OR 5.44, P < 0.001). CONCLUSIONS: Penicillins are the most frequently used antibiotics among school children. The results of this study demonstrate that antibiotic consumption is linked with higher carriage rates and resistance rates of respiratory tract pathogens. Therefore, rational use of antibiotics could prevent the emergence and spread of resistant bacteria.

Parental Knowledge and Awareness Linked to Antibiotic Use and Resistance: Comparison of Urban and Rural Population in Croatia.

Purpose: To investigate the differences in parental knowledge, attitudes, and practice about antibiotic use and resistance among the urban and rural populations in Croatia. Materials and Methods: A cross-sectional study based on a structured questionnaire was distributed to 1,000 parents of children attending 11 elementary schools of Primorsko-Goranska County in 2017. Results: The overall response rate was 65.1% (651/1,000)-50.6% (253/500) in urban and 79.6% (398/500) in rural population. Urban parents had a higher overall knowledge about antibiotics (p < 0.001), while urban and rural parents held mostly similar knowledge and attitudes related to antibiotic use when split into individual statements (p > 0.05). Age, education, income, work, and family member working in a health-related field were significantly related with the overall knowledge about antibiotics. In the previous year, 28.2% of children and 28.9% of parents reported using an antibiotic. Croatian parents had a high level of trust in doctors' antibiotic-prescribing practices (96.3% reported trusting the doctors' decision not to prescribe antibiotics, 93.5% to prescribe antibiotics) and high awareness of antimicrobial resistance (94.8%). The public's misconception regarding the terms "bacteria" and "virus" was found in 15.5% of parents. The source of information about antimicrobial resistance was television (60.4%), internet (57.1%), newspapers (44.2%), and medical professionals (30.9%). Conclusions: Although the knowledge about antibiotics was higher in urban parents, it was not reflected on the level of antibiotic consumption. There are indications that medical professionals should be more involved in communicating the problems of antibiotic use and resistance to patients.

Health Status, Lifestyle, Use of Health Services, Social Capital and Life Satisfaction As Predictors of Mental Health - Comparative Analysis of Women that Receive and Do not Receive Public Assistance in Croatia.

BACKGROUND: The connection between socio-economic status and health is documented, yet not fully understood. The goal of this research was to analyze the relationship between socio-economic status, lifestyle and health status, availability of health-care, social capital, and satisfaction with life. SUBJECTS AND METHODS: Subjects were 1117 women aged 25-65 years divided in two groups. Group 1 consisted of women who receive public assistance (N1=591), while Group 2 consisted of women who do not (N2=526). The sample was stratified by random choice into multiple stages based on six regions of Croatia, residential area size, and the age of respondents. Visiting nurses surveyed the deprived population, while in Group 2 self-interviewing was conducted. A questionnaire entitled "Inequalities in health" was used. The respondents participated in this research voluntarily and anonymously. RESULTS: Socially deprived women consume spirits and wine more often (p<0.001). There is no difference between groups regarding tobacco consummation. Working women perform significantly less strenuous physical tasks (p<0.001). Deprived women are significantly less engaged in physical activities (p<0.001). Health conditions in deprived women more commonly limit their physical activity (p<0.001). There is a significant difference in utilization of health-care among groups (p<0.001). Younger women who are married, with a higher number of household members, a larger income, and with higher education are generally more satisfied with life (p<0.001). Although deprived women are significantly less satisfied with their lives, feel less free, are less physically active, and less likely to consume spirits or beer, they are significantly happier than working women (p<0.001). CONCLUSIONS: Personal health status and lifestyle, access to health-care services, and life satisfaction have a high importance as predictors and protective factors of mental health in women - recipients of state-provided financial welfare.

Associations between Adherence to the Mediterranean Diet and Lifestyle Assessed with the MEDLIFE Index among the Working Population.

The adherence to the Mediterranean diet is beneficial for cardiovascular diseases prevention. The study aim is to use Mediterranean lifestyle (MEDLIFE) questionnaire for estimation of Mediterranean lifestyle habits among the working population and to establish MEDLIFE score correlation with the risk factors for cardiovascular diseases. In the study has participated 366 workers from Croatia, which fulfilled MEDLIFE and validated food frequency questionnaire (FFQ) questionnaire. The multivariate logistic regression was performed to evaluate the association between MEDLIFE index, workers' obesity and cardiovascular diseases risk. The lowest adherence to Mediterranean lifestyle was associated to younger, low education, body fat above acceptable ranges and unfavorable lipid profile. Significant association to Mediterranean lifestyle was more among women (p = 0.002), middle aged (p = 0.02), highly physically active (p = 0.009) and those who play collective sports >2 h/w (p = 0.001), having body fat within acceptable range (p = 0.003), total cholesterol less (p = 0.03) and high-density lipoproteins (HDL-C) (p = 0.04) more than recommended. Inverse significant association was for high educational level (p = 0.02). The Mediterranean lifestyle adherence is associated to lower risk factors for cardiovascular diseases among studied working population. MEDLIFE index revealed that physical activity and conviviality are better ingrained among younger population but not the Mediterranean diet.

Epidemiological study of visual functions--refractive errors, macular degeneration and glaucoma in children in the Karst area of Opatija.

Authors of earlier studies examined the epidemiological characteristics of certain eye diseases: age-related macular degeneration (AMD), refractive errors and glaucoma in the area of Primorsho-goranska County (the island of Rab, Novi Vinodolshi and Delnice). It was found that the occurrence of AMD is most common on the island of Rab, followed by Novi Vinodolski and it is least common in Gorshi Kotar. This fact is associated with the intensity of solar radiation in the UV-A and UV-B fields. The highest percentage of the occurrence of glaucoma was also identified on the island of Rab. In comparison to this study, it was found that in the karst area of Opatija (Mune, Brgud, Zejane, Bresca, Zvonece, Pasjak, Sapjane and Zaluki) there is a very high incidence of glaucoma (27% suspected and 7% diagnosed glaucoma) within the indigenous population. Glaucoma does not appear among children whose parents migrated to the karst area of Opatija. Refractive errors are far less common among children of indigenous population than among the children whose parents migrated to this area. The occurrence ofAMD was not found in any child that was born and lives in this area, regardless of whether their parents are indigenous or not. This statement is very important because it confirms author's earlier statement which claims that at low exposure to solar UV-A and UV-B there is no occurrence of AMD.

Perception of stress, depression, hypertension and myocardial infarction as predictors of adherence to hypertension drug treatment.

This survey was performed to determine the relationship between the adherence to hypertension drug treatment and the perception of stress, depression, hypertension, and myocardial infarction. 300 patients with uncomplicated hyperten- sion from Rijeka, Croatia, were included (131 women, 169 men, mean age 53.5 years). Adherence to hypertension drug treatment as criterion, and the perception of stress, depression hypertension and myocardial infarction as prediclors were determined by self-assessment. Collected data were analysed using factor analysis, regression analysis, Kolmogorov-Smirnov test, chi2-test and t-test. The statistical significance was set at a probability rate of less than 5% (p < 0.05). 45.09% of women (p=0.479), and 64.08% of men (p = 0.032) were motivated to take antihypertensives. 55.79% of women (p = 0.382) and 64.78% of men (p = 0.028) had sufficient knowledge about drug treatment of hypertension. The positive predictors of motivation for taking antihypertensives were physiological disturbances and perceived potency of hypertension and the negative were perceived helplessness in stress control and negative thoughts and emotions. The positive predictors of knowledge about taking antihypertensives were perceived helplessness in stress control, perceived potency of hypertension and myocardial infarction and the negative predictors were perceived self-efficacy in stress control, physiological disturbances and evaluation of hypertension. Both the motivation as well as the knowledge about taking antihypertensives should be improved, especially in women. The perception of stress, depression, hypertension and myocardial infarction can be used to predict adherence to hypertension drug treatment.

Refractive errors in children of Primorsko-Goranska County--epidemiological study.

Three institutions: Teaching Institute of Public Health of Primorsko-Goranska County, Croatian Association "Albert Einstein" and Eye Clinic "Dr B.Vojnikovid" Rijeka agreed a five-year project to study children's health status of vision at Primorsko-Goranska County. Main task was the study of damage of vision in children due to prolonged sun exposure. Examination were conducted on a three locations, with the assumption of varying insolation: Island of Rab, Novi Vinodolski and Delnice. The study included children aged between 9 to 14 years. The study included 189 children. Except routine ophthalmologic examinations in addition were preformed biomicroscopic examination of the anterior segment of the eye, intraocular pressure and eye fundus. In a certain number of children with suspect of a specific disease (macular degeneration, glaucoma and refractive error), additional tests were performed: glaucoma treatment, field of vision, optical coherence tomography (OCT) of the eye and detailed eye refraction. This study specifically addressed of refractive error at that three different regions. It was found that the most difficult situation was at the Novi Vinodolski where as many as 40% of children have a refractive error, followed by Island of Rab with 17% and Delnice with 9%. Especially like to mention that on the island of Rab, 50-60 years ago, it was a very rare occurrence of some refractive errors in children.

Evaluation of equivalence between different methods for enumeration of fecal indicator bacteria before and after adoption of the new Bathing Water Directive and risk assessment of pollution.

The quality of bathing water is of considerable public importance due to the possibility of fecal contamination. In 2009, Croatia implemented the new European Bathing Water Directive (BWD, 2006/7/EC) establishing stricter microbiological standards for new parameters with new reference methods. This study aims to evaluate the equivalence of different methods according to the old and revised BWD and to provide the possibility of data comparison. Furthermore, the directive requires the establishment of the bathing water profile (BWP) for pollution risk assessment. The estimation of consistency of pollution risk assessment with obtained microbiological results was also performed. Six marine beaches of the Municipality of Rijeka (Croatia) were examined during the 2009 season. Statistical analysis showed equivalence between determination methods for fecal contamination indicators. Based on the current water classification results, the need for correction of estimated pollution risks and recommendations for inclusion of historical microbiological data during BWP enactment was noticed.

The responses of the hepatosomatic index (HSI), 7-ethoxyresorufin-O-deethylase (EROD) activity and glutathione-S-transferase (GST) activity in sea bass (Dicentrarchus labrax, Linnaeus 1758) caged at a polluted site: implications for their use in environmental risk assessment.

The present study investigates the response of three hepatic biomarkers in adult sea bass (Dicentrarchus labrax, Linnaeus 1758) caged at a wastewater outlet of an oil refinery with fish caged at a pristine site used as controls. The biomarkers that were investigated were the hepatosomatic index (HSI), 7-ethoxyresorufin-O-deethylase (EROD) activity and glutathione-S-transferase (GST) activity. In addition, we have measured the levels of polycyclic aromatic hydrocarbons (PAHs) and selected heavy metals (lead, cadmium, mercury, copper and zinc) in sediment samples at the polluted site. Although the polluted site had high environmental levels of PAHs and heavy metals, there was no difference in hepatic EROD activity and HSI between fish caged at the polluted site and controls. On the other hand, GST activity was significantly lower in fish caged at the polluted site compared to controls. Our results point out that the studied biomarkers have limited use in environmental risk assessment studies, at least when caged adult sea bass is used as the sentinel species and complex toxicant mixtures are involved.

[Retrospection on Hygiene Congress organisation in Opatija 1950]. / Osvrt na organizaciju kongresa higijenicara u Opatiji 1950. godine.

In years following to the second world war the convention organisator was faced with major technical difficulties due to universal shortage of resources. During the 50's, two conventions of health workers with topics related to preventive medicine were held in Opatija. On both occasion Sanitary Bureau of Rijeka (Institute of Hygiene) was the main organisator. Reviewing the correspondance in Institute of Hygiene archive, one can monitor the preparations for the congress. In order to meet the participant's expectation, director of Institute of Hygiene, requested from the local entrepreneur adequate supply of postcard and cigarettes.

Residues of lindane in adipose tissue of lambs and sheep from islands Krk and Cres.

Concentrations of lindane from adipose tissue i.e. omentum of sheep from homesteads of island Krk that were treated with lindane, were compared with the concentrations of lindane measured in adipose tissue of non treated sheep from husbandry cooperative on island Cres. Evaluation of food safety for this samples were estimated by comparing obtained results and maximum tolerance concentration (MTC) for lindane (2.0 and 0.2 mg/kg). In treated lambs from Krk measured concentrations of lindane were in range from 0.0038 to 1.8644 mg/kg, and in treated sheep from 0.0094 to 1.646 mg/kg. In control group of lambs and sheep concentration of lindane was < or = 0.0051 mg/kg. Estimation of food safety based on the results from this research are discussable, regarding the fact that MTC prescribed before year 2007 was 2.0 mg/kg. When the new law Regulations of MTC for residues of pesticides in food and animal feed (NN 119/07) came into the force prescribed MTC was 0.02 mg/kg. If food safety of sheep meat from Krk would be estimated based on latter, then 73.4% of samples of lamb meat and 80% of sheep meat could be proclaimed unsafe for human consumption. It is encouraging that after completion of using lindane in year 2005, levels of lindane in adipose tissue of lamb from Krk in year 2006 were under the concentration set by new legislate (< 0.02 mg/kg).

Ecotoxicological characterization of marine sediment in Kostrena coastal area.

Samples of marine sediment were taken on 4 selected sites close to the shipyard industry in Kostrena coastal area. Concentration of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals (Cu, Pb, Zn, Hg, Fe) were analyzed from chemical-analytical and toxicological aspect. For toxicity detection, the bacterial bioluminescence test (Vibrio fisheri) was used. Concentration of total PAHs varied in the range from 697 to 7807 microg/kg dry weight in marine sediments. The concentration of PCBs in sediment was 1.1 mg/kg dry weight. The highest concentrations of heavy metals were found at the station within the shipyard. PAH toxicity was not correlated with the toxic potential of sediments. The obtained results indicate a high degree of environmental risk, especially at stations within the shipyard, with the 54% possibility of toxic effects. Chemical determination of the concentration of conventional pollutants is not sufficient for assessing the quality of the marine environment and it is necessary to use other approaches in order to evaluate the biological impact.

Cancer and cardiovascular diseases nutrition knowledge and dietary intake of medical students.

The aims of this study were to determine medical students' knowledge regarding the association between dietary factors and the risk of cancer and cardiovascular diseases and to investigate if this knowledge has an impact on their dietary intakes. Three hundred and ninety medical students (males and females) were included in a study and grouped according to their daily fibre and fat intakes. For diet-disease knowledge, questions from the General Nutrition Knowledge Questionnaire for Adults were used and dietary assessment was done with Food Frequency Questionnaire. The obtained results showed that the students' diet-disease knowledge was generally inadequate. Higher level of diet-disease knowledge was among those with high dietary fibre intake, with slightly better scores for dietary factors and risk for cardiovascular diseases than the risk for cancer. Better diet-disease knowledge positively correlated with higher intake of fish (p = 0.027, p = 0.001) and vegetables (p = 0.019, p = 0.001) in high fibre groups of both gender, and in females additionally with fruit intake (p = 0.038, p = 0.007). A higher dietary fibre intake among studied students seems to be a factor that ensures lower obesity rates, lower intake of energy and lower consumption of coffee, sweets and alcoholic drinks. On the basis of the results of this study, it is clear that medical schools should provide in their nutrition programs the opportunity for students to learn about their own dietary and lifestyle behaviours, in order to more knowledgably and convincingly counsel their future patients.

Perception of arterial hypertension and myocardial infarction in hypertensive and normotensive men and women.

The research aims were to test perception of arterial hypertension and myocardial infarction in hypertensive and normotensive men and women as well as to test perception of arterial hypertension and myocardial infarction as predictors of blood pressure control in hypertensives. In the research 470 subjects of 4 general practices from Rijeka, Croatia participated, hypertensive group from the list of hypertensive patients without cardiovascular complications and other major chronic conditions, normotensive group from the list of patients without chronic conditions. Each group had 235 subjects, 128 men and 107 women. Perception of hypertension and myocardial infarction was measured as the result on semantic differential questionnaire. Factor analysis extracted evaluation, potency and activity factor. Blood pressure control was interpreteted on the five degrees scale. Statistical significance was defined under 5% (p < 0.05). Hypertensive subjects perceived hypertension as less negative and more active, while myocardial infarction was perceived as more potent term than by normotensives. Women perceived myocardial infarction as less negative, and less potent term than men. Both groups perceived myocardial infarction as more negative, potent and active term than hypertension. Normotensive women evaluated hypertension as more negative, and perceived myocardial infarction as less potent than other subjects. Well-controlled hypertension was correlated with a lower potency of hypertension and lower activity of myocardial infarction. Both conditions are perceived as more "male" diseases. As perception of hypertension and myocardial infarction is correlated with blood pressure regulation in hypertensives, and hypertension is major risk factor for myocardial infarction, family doctors should put additional effort in changing perception of cardiovascular diseases in their patients, especially in women.

Genotoxicity of marine sediments in the fish hepatoma cell line PLHC-1 as assessed by the Comet assay.

The main goal of this study was to test the usefulness of the Comet assay in the PLHC-1 hepatoma fish cell line as a tool for detecting the presence of genotoxic compounds in contaminated marine sediments. The system has been tested using both model chemicals (benzo[a]pyrene (B[a]P) and ethyl methanesulfonate (EMS)) and extracts of sediment samples obtained with solvent dichloromethane/methanol. For all of the analysed sediment extracts as well as for the model chemicals a concentration dependent genotoxic effect was observed. The sediment with the highest observed genotoxic potential was additionally extracted using various solvents in order to test which class of compounds, according to their polarity, is most responsible for the observed genotoxic effect. Non-polar solvents (cyclohexane and dichloromethane) yielded stronger genotoxic effect but the highest level of DNA damage was determined after exposure to sediment extract obtained with the solvent mixture dichloromethane/methanol which extracts a wide range of contaminants. Our results indicate that the PLHC-1 cell line is a suitable in vitro model in sediment genotoxicity assessment and encourage the use of fish cell lines as versatile tools in ecogenotoxicology.

Polycyclic aromatic hydrocarbons and heavy metals in Kostrena coastal area.

The aim of this study was to determine pollution by polycyclic aromatic hydrocarbons (PAH) and heavy metals in seawater and sediment in Kostrena coastal area, as well as their toxicity using bioluminescence based tests. Total PAH concentration in seawater ranged 1.7-155.3 ng/L. The share of carcinogenetic PAH was relatively high, ranging 22-48.3%. Nickel concentrations in seawater were beyond detection limits (< 0.1 microg/L), vanadium concentrations ranged 0.66-1.96 microg/L, chrome concentrations were beyond detection limits, and copper concentrations were also beyond detection limits or extremely low (up to 0.32 microg/L). EC50 values in seawater ranged 23.80-90.90 ng/L. Correlation between total PAH concentration and toxicity of seawater showed strong connection between them (r = 0.9579). Total PAH concentration in marine sediment ranged 58.02-1116 microg/kg dry weight (d.w.). The share of carcinogenetic PAH was extremely high ranging 10-53%. Nickel concentrations in marine sediment ranged 8-24 mg/kg d.w., vanadium concentrations ranged 24-42 mg/kg d.w., chrome concentrations ranged 11-19 mg/kg d.w., and copper concentrations ranged 7-25 mg/kg d.w. EC50 values in marine sediment ranged 818-4596 microg/kg d.w. Correlation between total PAH concentration and toxicity of marine sediment showed weak connection between them (r = 0.2590). Previous studies of seawater samples from areas of the Adriatic sea under the direct influence of oil industry did not include concentrations of heavy metals, which makes our study the first to present such comprehensive results. Our results point out the need for further evaluations and following of marine environment pollution and its consequences on living organisms and marine ecosystem in whole.

Monitoring the presence of genetically modified food on the market of the Republic of Croatia.

From the beginning of the human race people have been applying different methods to change the genetic material of either plants or animals in order to increase their yield as well as to improve the quality and quantity of food. Genetically modified organism (GMO) means an organism in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination. Analysing the presence of GMO in food is done by detecting the presence of either specific DNA sequences inserted in the genome of transgenic organism, or detecting proteins as a result of the expression of the inserted DNA. In this work food testing for the presence of genetically modified organisms was conducted during the period from 2004 to 2007 in the GMO laboratory of the Croatian National Institute of Public Health. According to the regulations, among the samples in which the presence of GMO was detected, all those which had more than 0.9% of GMO content were either rejected from the border or removed from the market, because such GM food has to be appropriately labelled. Among the food samples which were analysed in 2004: 127 (2.37%) of a total of 1226 samples contained more than 0.9% of GMOs; in 2005 there was only one in 512 (0.20%) samples in total; in 2006 there were 4 out of 404 samples (0.99%), and in 2007: 7 of a total of 655 samples (1.07%) had GMO content above the allowed threshold of 0.9%.

Immunomodulation of cell-mediated cytotoxicity after chronic exposure to vapors.

Cytotoxic T lymphocytes and natural killer (NK) cells represent two major types of immune cytotoxic cells that have a crucial role in innate immunity. Urban air pollution represents a common and difficult problem in the majority of metropolises, which contain high levels of traffic congestion generating great amounts of genotoxic substances. Diesel exhaust particles act as adjuvant in the immune response and may lead to the enhancement of proinflammatory and proallergic response without exposure to allergen. The activities of diesel exhaust particles (DEPs) and their benzene extracts may cause the increasing respiratory mortality and morbidity. Since the lung inflammatory response to DEPs is compartmentalized, the importance of finding the suppressive substances, whose function will be to elucidate the DEPs effects, is fundamental. The objectives of this study were to examine the NK-mediated cytotoxicity against human NK-sensitive target, as well as the values of BTEX (benzene, toluene, ethylbenzene and xylene isomers) in urine in people from industrial area and those from rural fields. NK-mediated cytotoxicity of peripheral blood mononuclear cells was investigated against human NK-sensitive K-562 tumor cell line. BTEX were analyzed from urine by a HS-SPME method. All exposed individuals were demonstrated the significant decreased of NK-mediated cytotoxicity against human NK-sensitive target and increasing BTEX values. BTEX-analyze in urine and determination of NK-mediated cytotoxicity, as an important methods represent good markers in environmental monitoring, which contribute to the better protection of air pollutants and quicker diagnosis of environmental diseases.

Determination of enzyme matrix metalloproteinases-9 and immune status as indicators of development of the environmental diseases.

The majority of environmental diseases are multifactorial airway illnesses, including genetic background and exposure to different kind of airborne irritants and allergens. Altered lifestyle and changes in environmental exposures contribute to the occurring of these diseases. The term of environmental illnesses includes the disease primarily caused by pollution of air and water, chemical and physical agents, radiation, contaminated food and direct contact with the toxins we are exposed to natural and/or working environment. The members of the matrix metalloproteinase (MMP) family are involved in the pathogenesis of COPD. MMPs comprise a large family of structurally related zinc metalloendopeptidases with different substrate specificities and possibilities to degrade protein constituents of the extracellular matrix. We investigated immunological status and level of MMP-9 in workers occupationally exposed to volatile aromatic hydrocarbons compared to urban residents and rural areas. The phenotypic profiles of peripheral blood lymphocytes were done by flow cytometry. The method of enzyme immunoassay (ELISA) was used to determine enzyme expression of matrix metalloproteinase-9 (MMP-9). The occupationally exposed group had a significantly elevated level of enzyme MMP-9 in the urine, accompanied with augmentation of cells of innate immunity in peripheral blood, which could contribute to the monitoring, early detection of environmental diseases and consequent earlier and more effective treatment.