Jiří Jarkovský

Addition of Spironolactone in Patients With Resistant Arterial Hypertension (ASPIRANT) A Randomized, Double-Blind, Placebo-Controlled Trial

There is currently limited data on which drug should be used to improve blood pressure (BP) control in patients with resistant hypertension. This study was designed to assess the effect of the addition of 25 mg of spironolactone on BP in patients with resistant arterial hypertension. Patients with office systolic BP >140 mm Hg or diastolic BP >90 mm Hg despite treatment with at least 3 antihypertensive drugs, including a diuretic, were enrolled in this double-blind, placebo-controlled, multicenter trial. One hundred seventeen patients were randomly assigned to receive spironolactone (n=59) or a placebo (n=58) as an add-on to their antihypertensive medication, by the method of simple randomization. Analyses were done with 111 patients (55 in the spironolactone and 56 in the placebo groups). At 8 weeks, the primary end points, a difference in mean fall of BP on daytime ambulatory BP monitoring (ABPM), between the groups was −5.4 mm Hg (95%CI −10.0; −0.8) for systolic BP (P=0.024) and −1.0 mm Hg (95% CI −4.0; 2.0) for diastolic BP (P=0.358). The APBM nighttime systolic, 24-hour ABPM systolic, and office systolic BP values were significantly decreased by spironolactone (difference of −8.6, −9.8, and −6.5 mm Hg; P=0.011, 0.004, and 0.011), whereas the fall of the respective diastolic BP values was not significant (−3.0, −1.0, and −2.5 mm Hg; P=0.079, 0.405, and 0.079). The adverse events in both groups were comparable. In conclusion, spironolactone is an effective drug for lowering systolic BP in patients with resistant arterial hypertension.

Soil burdens of persistent organic pollutants - their levels, fate and risk. Part I. Variation of concentration ranges according to different soil uses and locations.

Detailed soil screening data from the Czech Republic as a typical Central European country are presented here. Determination of a wide selection of organic and inorganic pollutants as well as an assessment of specific soil parameters allowed us to study the soil contamination in relation to the land use and soil properties. While HCHs and HCB were found at highest levels in arable soils, the higher concentrations of PCDDs/Fs, PCBs, PAHs and DDTs were observed in high altitude forest soils. Concentrations of these compounds strongly correlated with the soil organic carbon content. Several possible reasons have been suggested for the observed higher concentrations in mountain forest soils but the impact of each of these influencing factors remains to be identified. An inventory of the soil contamination is needed as a first step in our effort to estimate an extent to which the secondary sources contribute to the enhanced atmospheric levels of POPs.

Assessment of Organic Pollution Effect Considering Differences between Lotic and Lentic Stream Habitats

Based on the requirements of the Water Framework Directive, a macroinvertebrate-based assessment system to evaluate the ecological quality of streams has been developed by AQEM project consortium. In the Czech Republic the impact of organic pollution was principal pressure studied, but some morphological degradation of some sampling sites could not be avoided. A multimetric assessment system for three stream types was developed. Detrended Correspondence Analysis was used for the detection of the response of macroinvertebrate communities to the gradient of organic degradation. Significant relationships between abiotic (BOD, TOC, nutrients) and biotic (saprobic index, ASPT) indicators of organic enrichment/eutrophication were identified. Separate storage of the riffle and pool components of each multi-habitat sample allowed differences between these habitats to be com- pared in context of the metrics applied in the assessment system. Lotic and lentic habitats differed in taxonomic composition, ecological traits and biotic indices. The separate assessment of the riffle and pool parts of samples provides additional useful information when combined effects of organic pollution and morphological degradation are to be considered.

Trends in background levels of persistent organic pollutants at Kosetice observatory, Czech Republic.

A multimedia sampling of ambient air, wet deposition, surface water, sediment, soil and biota has been performed at Kosetice background observatory in the southern Czech Republic since 1988. An integrated monitoring approach was applied to assess the current state, anthropogenic impacts, and possible future changes of terrestrial and freshwater environments. Average PCB concentrations in the individual matrices calculated from ten years of sampling on multiple sites varied between 2 ng g(-1) in sediment and 7 ng g(-1) in soil or moss. DDT concentrations were lower in moss and needles (2 ng g(-1) and 4 ng g(-1), respectively) than in sediment (11 ng g(-1)) and soil (20 ng g(-1)), while the HCH level was higher in moss and needles (5 ng g(-1) and 6 ng g(-1), respectively) than in soil or sediment (1 ng g(-1) and 2 ng g(-1), respectively). The highest average level of PAHs was found in soil (600 ng g(-1)), while it was lower in needles (230 ng g(-1)), moss (210 ng g(-1)) or sediment (210 ng g(-1)). Time related trends of concentration levels of persistent organic pollutants in all matrices were investigated. Moss and needle trend patterns resembled those of the ambient air, showing a slight concentration decrease of all compounds, except for hexachlorobenzene. The soil, water and sediment concentrations showed a similar decrease of PAHs, PCBs, and HCHs, but there was no clear trend for DDTs and HCB.

The PERLA system in the Czech Republic: a multivariate approach for assessing the ecological status of running waters

The assessment of running water quality has a long tradition in the Czech Republic, but in the past it focused on the evaluation of organic pollution using the saprobic system. Considering the modern trends of stream ecological status evaluation in water management a new assessment system named PERLA was developed. The system is a complex of biological methods of ecological status assessment of running waters and connected activities in the Czech Republic. It involves 300 reference sites with respective biotic and abiotic data and a prediction model using a newly developed software HOBENT. The model generally follows the published mathematical principles of RIVPACS and represents the site specific and stressor non-specific approaches. The HOBENT software allows the prediction of the target assemblage of benthic macroinvertebrates for any site based on a set of environmental variables (latitude, longitude, distance from source, altitude, slope, catchment area, and stream order) which characterise the site. The predicted assemblage can be compared with the fauna observed at the same site. The comparison makes it possible to evaluate the extent of disturbance, expressed by index B. The model allows to evaluate spring, summer, and autumn seasonal data of the majority of wadable streams in the Czech Republic. The practical application of the PERLA system has started in 2001.

Parasite communities of freshwater fish under flood conditions.

Abstract. The parasite communities of the freshwater fish species roach (Rutilus rutilus), bitterling (Rhodeus sericeus amarus) and perch (Percafluviatilis) were investigated during a major flood. Differences in parasite community structure due to changes in the host environment were expected. We therefore tested for differences in parasite species abundance and diversity as a consequence of the flood. Potential changes in parasite community structure due to seasonal influences were also considered. We found differences in the composition of parasite species and the proportion of ecto- and endoparasites in the three host species. The parasite community of roach was richer in species than those of perch and bitterling. Roach harboured more ectoparasite than endoparasite species. Parasite communities of both perch and bitterling were richer in endoparasite species and had a lower number of ectoparasite species. Parasite communities of the three different host species responded in different ways to environmental change. Considering all parasite species, no effect of either season or flood was found on the species diversity in either perch or bitterling. The flood caused an increase in the parasite species diversity in roach. For roach and perch, higher parasite abundance were found before the flood compared to after flood. However, no difference was found between the different seasons after the flood. By contrast, bitterling had the highest parasite abundance in the season after the flood. When analysing ecto- and endoparasites separately, a decrease in ectoparasite abundance was found in roach and perch, probably related to the flood, and due mainly to a decrease in monogenean species. A significant increase in the abundance of endoparasites was found in perch. In bitterling, the post-flood increase in ectoparasites was due to a high abundance of Gyrodactylus spp. We conclude that the parasite communities of the three fish species responded in different ways to the flood. This may be due to changes in the host life history strategy or in the immune response in conditions of stress.

Effects of fungicides mancozeb and dinocap on carbon and nitrogen mineralization in soils.

In our study, effects of fungicides mancozeb and dinocap on C and N mineralization were measured in arable and grassland soil. The soils were treated with these fungicides at the application and 10 times lower doses and then incubated at 20 degrees C for 2 weeks. Carbon mineralization (basal and substrate-induced respiration) and nitrogen mineralization (potential ammonification and nitrification) were evaluated 1 and 14 days after the treatment. After 14 days, ammonification was decreased to 48% and 83% at dinocap application dose in arable and grassland soil, respectively. Application dose of mancozeb caused significant decrease of nitrification to 11.2% and 5.6% in arable and grassland soil, respectively. Basal respiration and substrate-induced growth were rather stimulated by fungicides, especially at lower application doses. To conclude, potential risk may exist to soil microorganisms and their activities in soils treated routinely by mancozeb or dinocap.

Variability of soil microbial properties: effects of sampling,handling and storage

We investigated the effect of soil spatial variability within the sampling site scale, the effects of sample sieving (1, 2 and 4mm), and storage conditions up to 32 weeks (wet at 4 degrees C, -20 degrees C and air dried) on microbial biomass C, respiration, ammonification and nitrification activities in arable, grassland and forest soil. In general, all results were dependent on soil type. Arable soil showed the highest spatial variability, followed by grassland and forest soil. Sieving did not cause large differences; however, higher biomass C and respiration activity were observed in the 1mm than in the 4mm fraction. Storage at 4 degrees C seemed to be the most appropriate up to 8 weeks showing only minor changes of microbial parameters. Freezing of soils resulted in large increase of respiration. Dried storage indicated disruption of microbial communities even after 2 weeks.

Uric acid, allopurinol therapy, and mortality in patients with acute heart failure—results of the Acute HEart FAilure Database registry

STUDY OBJECTIVE The aim of this study was to explore the prognostic role of serum uric acid (UA) measurement in the hospital and long-term mortality assessment in subjects with acute heart failure (AHF) from the Acute HEart FAilure Database registry (AHEAD). The AHEAD registry comprised 4153 patients with AHF syndromes hospitalized at the AHEAD participating centers. PATIENTS AND METHODS The study included 1255 patients who were admitted to the AHEAD participating centers with acute decompensated chronic heart failure, de novo heart failure, or cardiogenic shock between September 2006 and October 2009 and who had information about serum UA concentration available at the time of hospital admission. The hospital and long-term mortality was followed using the centralized database of the Ministry of Health, Czech Republic. The mean age of the cohort was 73.4 years, the female population represented 43%, the median hospital stay was 8 days, and the mean hospital mortality was 7.6%. RESULTS The median UA concentration of the patients with AHF was 432 μmol/L (7.26 mg/dL), the median estimated glomerular filtration rate (eGFR) was 49.0 mL/min, and N-terminal pro-brain natriuretic peptide level was 5510 pg/mL. Among other laboratory variables, UA concentration greater than 515 μmol/L (8.67 mg/dL) was associated with increased hospital mortality (P < .001), as well as eGFR less than 30 mL/min (P < .001), Na 135 mmol/L or less, and positive troponin. Uric acid concentration greater than 500 μmol/L (8.41 mg/dL) was associated with increased long-term mortality (P < .001), followed by eGFR less than 30 mL/min (P < .001), Na 135 mmol/L or less, and hemoglobin level lower than 130 g/L (P < .001). The 1-year survival rate of patients discharged from hospital (n = 1159) was 75.6%, and the 2-year rate was 66.8%. Survival of patients treated with allopurinol for hyperuricemia was significantly lower compared with untreated subjects (70.1 vs 77.2 for 1-year survival and 60.3 vs 68.5 for 2-year survival). CONCLUSION In patients with AHF, increased UA levels and documented allopurinol therapy for hyperuricemia were associated with increased hospital and long-term mortality. Allopurinol therapy is not a cause but the identifier of the subjects at risk.

Organic Pollutant Contamination of the River Tichá Orlice as Assessed by Biochemical Markers

·iroka Z. , J . Kri j t , T. Randak, Z. Svobodova, G. Pe‰kova, J . Fuksa, J . Haj‰ lova, J . Jarkovsk , M. Janska: Organic Pollutant Contamination of the River Elbe as Assessed by Biochemical Markers. Acta Vet Brno 2005, 74: 293-303. The aim of the study was to assess contamination of the River Elbe basin using selected biochemical markers. Biochemical markers selected were enzymes of the first stage of xenobiotic transformation, namely cytochrome P450 (CYP 450) and ethoxyresorufin-O-deethylase (EROD). The results were correlated with the most important inducers of the enzymes, i.e. polychlorinated biphenyls (PCB) concentrations in muscle tissue of fish, polyaromatic hydrocarbons (PAH) values in bottom sediments and 1-hydroxypyrene (1-OHPY) values in fish bile (terminal metabolite of PAH, or, rather, of one of them, i.e. pyrene), which were determined during the chemical monitoring of the River Elbe basin. The indicator species selected was chub (Leuciscus cephalus L.), which was captured at ten locations in the River Elbe basin. A comparison between the EROD activity and the CYP 450 content along the longitudinal profile of the Elbe showed a significant correlation at the level of significance of p < 0.05. The highest EROD activity levels in the liver were ascertained in Zelain (341 pmol·min-1·mg-1), Valy (263.2 pmol·min-1·mg-1) and Lysa nad Labem (179.17 pmol·min-1·mg-1). In Blanice (control location), EROD activity was significantly lower than in any of the other locations studied (p < 0.05). The study failed to produce an unambiguous proof of any correlations between detoxification enzyme activity (CYP 450 and EROD) in the liver and their two important inducers (PCB and PAHs). The possibility that other substances causing activation or inhibition of detoxification enzymes were in play is also discussed. Cytochrome P450, EROD, Leuciscus cephalus L., liver, PCB, PAH, 1-hydroxypyrene The River Elbe is one of the most important European rivers (total length 1103.5 km). Its extensive basin of a total of 148 268 km2 lies on the territory of two countries, i.e. the Czech Republic (51 336 km2) and Germany (96 932 km2). Intensive research of the Czech and German reaches of the Elbe started in 1991 under the Elbe I (1991 1994) project, and continued with the Elbe II (1995 1998) and Elbe III (1999 2002) projects. In those projects, large quantities of data regarding sources of pollution, chemical monitoring of hazardous substances in various components of the aquatic environment, water quality, etc. were collected and evaluated (Nesmurak 1994; BlaIkova et al. 1998; BlaIkova 2002). To enhance the relevance of results obtained by chemical monitoring, it is, however, also ACTA VET. BRNO 2005, 74: 293–303 Address for correspondence: ·iroka Zuzana Department of Public Veterinary Health and Toxicology University of Veterinary and Pharmaceutical Sciences Brno Palackeho 1-3, 612 42 Brno, Czech Republic Phone: +420 541 562 780 E-mail: sirokaz@vfu.cz http://www.vfu.cz/acta-vet/actavet.htm necessary to assess the effects of anthropogenic pollution of the aquatic environment on fish. One of possible ways of assessing such effects is the use of biochemical markers of contamination. They are measurable biochemical parameters responding usually to substances with the same mechanism of toxic effect. That means they are not, with some exceptions, specific for individual xenobiotic substances. Their advantage lies in their ability to provide comprehensive information on the effects of pollution, i.e. to reflect synergic or antagonistic effects of individual components contributing to pollution. In 2003, the Elbe IV Project was started. For reasons mentioned above, chemical monitoring in fish was complemented with assay of biochemical contamination markers. Attention focused primarily on the enzymes of the first phase of xenobiotics conversion, i.e. cytochrome P450 and ethoxyresorufin-O-deethylase (EROD). Cytochrome P450 is an important biochemical marker of surface water contamination with some industrial and agricultural pollutants (Stegeman and Lech 1991). It is now believed that the most useful is the 1A family of cytochrome P450 (Machala et al. 1997; Schlenk and Di Giul io 2002). The most potent inducers for that isoform are substances from the groups of polychlorinated biphenyls (PCB), polycyclic aromatic hydrocarbons (PAH), nitrated polycyclic aromatic hydrocarbons (NPAH) and dioxins (e.g. 2, 3, 7, 8 TCDD) (White et al. 1997; Nilsen et al. 1998; Jung et al. 2001; Schlenk and Di Giul io 2002). On the other hand, chronic exposure to these contaminants can cause a lack of CYP1A induction response (Brammell et al. 2004), and also assessment of CYP1A at the time of spawning can influence its level, because estrogens can decrease CYP1A induction (Elskus 2004). Male fish seem to be more sensitive to PAH and PCB than female fish (McArdle et al. 2004). The induction of the CYP1A family is mediated by the aryl hydrocarbon receptor (AhR) (Bil l iard 2002). Following its interaction with xenobiotic substances, it is carried to the nucleus where it is the cause of enhanced expression of genes for CYP1A and, subsequently, of increased synthesis of cytochrome proteins. The potential toxicity of pollutants depends on their affinity to the AhR. The CYP1As are also responsible for the metabolic activation of most of the known promutagens and procarcinogens, and its elevated levels are responsible for such negative effects as cocarcinogenesis, immunotoxicity and reproduction disorders (Lewis et al. 2003; Carlson et al. 2004). The model CYP1A activity is the enzyme ethoxyresorufin-O-deethylase (EROD), with its ability to convert substrates to products demonstrating fluorescence, which can then be measured. This enzyme is an important biochemical marker of contamination. The aim of the study presented here was to use the assessment of biochemical markers cytochrome P450 and EROD in the livers of the indicator fish species (Leuciscus cephalus L.) to evaluate contamination levels in various locations within the River Elbe basin. Results of chemical monitoring relevant for the above contamination markers are also outlined and correlated in the paper. They were PCB concentrations in chub muscle tissues, concentrations of 1-hydroxypyrene (1-OHPY) in chub bile samples (i.e. the final metabolite of PAHs, or rather of pyrene), and PAH concentrations in bottom sediments in the locations studied. Materials and Methods Animals and Sampling The chub (Leuciscus cephalus L.) was selected as the most suitable indicator species. The chub is a common freshwater cyprinid species that inhabits both clean and polluted rivers (Baru‰ et al. 1995). The fish were captured with the use of a diesel-electric generator in 10 locations in the River Elbe basin. The locations studied were Podoli and Zelain at the River Vltava, a tributary to the Elbe, and Verdek, Numaice, Valy, Lysa nad Labem, Obfiistvi, Duain and Hfiensko along the River Elbe. The control location was upstream of the Husinec water reservoir at the river Blanice in the Vltava basin (Fig. 1). The fish were captured in July 2003 (average water temperature 21.4 °C). In the control location, fish were captured in September 2003 (water temperature 15 °C). In each location, eight chub (both males and females) were captured (except in Lysa nad Labem where only 3 chub were captured). The chub 294