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Abstract Background Diplomonads are anaerobic flagellates classified within Metamonada. They contain both host-associated commensals and parasites that reside in the intestinal tracts of animals, including humans (e.g., Giardia intestinalis), as well as free-living representatives that inhabit freshwater and marine anoxic sediments (e.g., Hexamita inflata). The evolutionary trajectories within this group are particularly unusual as the free-living taxa appear to be nested within a clade of host-associated species, suggesting a reversal from host-dependence to a secondarily free-living lifestyle. This is thought to be an exceedingly rare event as parasites often lose genes for metabolic pathways that are essential to a free-living life strategy, as they become increasingly reliant on their host for nutrients and metabolites. To revert to a free-living lifestyle would require the reconstruction of numerous metabolic pathways. All previous studies of diplomonad evolution suffered from either low taxon sampling, low gene sampling, or both, especially among free-living diplomonads, which has weakened the phylogenetic resolution and hindered evolutionary insights into this fascinating transition. Results We sequenced transcriptomes from 1 host-associated and 13 free-living diplomonad isolates; expanding the genome scale data sampling for diplomonads by roughly threefold. Phylogenomic analyses clearly show that free-living diplomonads form several branches nested within endobiotic species. Moreover, the phylogenetic distribution of genes related to an endobiotic lifestyle suggest their acquisition at the root of diplomonads, while traces of these genes have been identified in free-living diplomonads as well. Based on these results, we propose an evolutionary scenario of ancestral and derived lifestyle transitions across diplomonads. Conclusions Free-living taxa form several clades nested within endobiotic taxa in our phylogenomic analyses, implying multiple transitions between free-living and endobiotic lifestyles. The evolutionary history of numerous virulence factors corroborates the inference of an endobiotic ancestry of diplomonads, suggesting that there have been several reversals to a free-living lifestyle. Regaining host independence may have been facilitated by a subset of laterally transferred genes. We conclude that the extant diversity of diplomonads has evolved from a non-specialized endobiont, with some taxa becoming highly specialized parasites, others becoming free-living, and some becoming capable of both free-living and endobiotic lifestyles.

ObjectivesCardiac arrhythmias predict poor outcome after myocardial infarction (MI). We studied if arrhythmia monitoring with an insertable cardiac monitor (ICM) can improve treatment and outcome.DesignBIO|GUARD-MI was a randomized, international open-label study with blinded outcome assessment.SettingTertiary care facilities monitored the arrhythmias, while the follow-up remained with primary care physicians.ParticipantsPatients after ST-elevation (STEMI) or non-ST-elevation MI with an ejection fraction >35% and a CHA2DS2-VASc score ≥4 (men) or ≥5 (women).InterventionsPatients were randomly assigned to receive or not receive an ICM in addition to standard post-MI treatment. Device-detected arrhythmias triggered immediate guideline recommended therapy changes via remote monitoring.Main outcome measuresMACE, defined as a composite of cardiovascular death or acute unscheduled hospitalization for cardiovascular causes.Results790 patients (mean age 71 years, 72% male, 51% non-STEMI) of planned 1,400 pts were enrolled and followed for a median of 31.6 months. At 2 years, 39.4% of the device group and 6.7% of the control group had their therapy adapted for an arrhythmia [hazard ratio (HR) = 5.9, P

Background and aimsAnalysis of mortality from the national health registries and data from a specific central registry dealing with the implantation of pacemakers (REPACE) in Czech patients.Methods and resultsRetrospective observational analysis of pacemakers’ implantation in all Czech patients [n = 82,791; 47,070 (56.9%) men, 75.9 ± 10.4 years old] between 2010 and 2021. Almost 114,000 pacemakers were implanted between 2010 and 2021, of which 27.9% were single-chamber, 67.4% were dual-chamber and 4.6% were biventricular. The annual number of implantations has been steadily increasing with a 6% annual decline in 2020 with increased mortality and reductions in care provided, likely related to COVID-19. The observed 5-year relative survival was 88.6% (overall survival 60.6%) and the 10-year relative survival was 75.9% (overall survival 32.7%). Causes of death 5ary according to the age of the patient. The highest difference 1n the reported numbers in the REPACE Registry did not exceed 2% in comparison with the National Register of Reimbursed Health Services.ConclusionThis study followed all Czech patients with pacemaker’s implantation in between 2010 and 2021. The annual number of 1mplantations has been steadily 1ncreasing. Patients with implanted pacemakers had a significantly higher mortality than the average population. Number of patients in the registry corresponded almost perfectly with the National Register of Reimbursed Health Services.

Abstract Fetal intracranial hemorrhage represents a rare event with an estimated prevalence of 1:10 000 pregnancies. We report a patient diagnosed prenatally with intracranial hemorrhage and ventriculomegaly carrying a novel, previously unreported, likely pathogenic variant in COL4A1. At the gestational age of 27 weeks, dilation of lateral ventricles was detected during a routine prenatal ultrasound scan, confirmed by prenatal MRI at 30 + 3 weeks of gestation. Prenatal examinations included amniocentesis with conventional G‐band karyotyping and arrayCGH, and maternal testing for TORCH and parvovirus B19 infections. Virtual gene panel based on whole‐exome sequencing data was performed postnatally. At the age of 2.5 months, the patient manifested epileptic seizures that remain difficult to control. Postnatal MRI showed partial thalamic fusion and polymicrogyria, in addition to severe enlargement of lateral ventricles, multiple deposits of hemosiderin in cerebral and cerebellar hemispheres, and thin optic nerve and chiasma. Virtual gene panel based on whole‐exome sequencing data led to a detection of a de novo previously unreported in‐frame deletion NM_001845.5:c.4688_4711del in COL4A1 located in the highly conserved NC1 domain initiating collagen helix assembly. The presented case lies one a more severe end of the COL4A1 mutation‐related disease spectrum, manifesting as fetal intracranial bleeding, malformation of cortical development, drug‐resistant epilepsy, and developmental delay.

Abstract Aims Fabry disease (FD) is a rare X‐linked genetic disorder caused by α‐galactosidase A (AGALA) deficiency. Whereas ‘classic’ variant has multisystemic manifestation, the more recently described ‘later‐onset’ variant is characterized by predominant cardiac involvement that often mimics hypertrophic cardiomyopathy (HCM). Methods and results Consecutive unrelated patients with HCM were screened for FD in 16 (out of 17) cardiac centres in the Czech Republic covering specialized cardiology care from June 2017 to December 2018. AGALA activity and globotriaosylsphingosine (lyso‐Gb3) levels were measured in all subjects using the dry blood spot method. FD was suspected in male patients with AGALA activity 3.5 ng/mL. Positive screening results were confirmed by genetic testing. We evaluated 589 patients (390 males, 66%) with HCM (mean maximal myocardial thickness 19.1 ± 4.3 mm). The average age was 58.4 ± 14.7 years. In total, 17 patients (11 males, 6 females) had a positive screening result, and subsequently, six of them (four males and two females) had a genetically confirmed pathogenic GLA mutation (total prevalence of 1.02%). Five of these patients were carrying the p.N215S mutation known to cause a typical later‐onset cardiac FD. Conclusions We confirmed the prevalence of FD repeatedly reported in previous screening programmes (approximately 1% irrespective of gender) in a non‐selected HCM population in Central Europe. Our findings advocate a routine screening for FD in all adult patients with HCM phenotype including both genders. The dry blood spot method used led to identification of clearly pathogenic variants.

An association between congestive heart failure (CHF) and chronic kidney disease (CKD) results in extremely poor patient survival rates. Previous studies have shown that increasing kidney epoxyeicosatrienoic acids (EETs) by blocking soluble epoxide hydrolase (sEH), an enzyme responsible for EETs degradation, improves the survival rate in CHF induced by aorto-caval fistula (ACF) and attenuates CKD progression. This prompted us to examine if sEH inhibitor treatment would improve the outcome if both experimental conditions are combined. Fawn-hooded hypertensive (FHH) rats, a genetic model showing early CKD development was employed, and CHF was induced by ACF. Treatment with an sEH inhibitor was initiated 4 weeks after ACF creation, in FHH and in fawn-hooded low-pressure (FHL) rats, a control strain without renal damage. The follow-up period was 20 weeks. We found that ACF FHH rats exhibited substantially lower survival rates (all the animals died by week 14) as compared with the 64% survival rate observed in ACF FHL rats. The former group showed pronounced albuminuria (almost 30-fold higher than in FHL) and reduced intrarenal EET concentrations. The sEH inhibitor treatment improved survival rate and distinctly reduced increases in albuminuria in ACF FHH and in ACF FHL rats, however, all the beneficial actions were more pronounced in the hypertensive strain. These data indicate that pharmacological blockade of sEH could be a novel therapeutic approach for the treatment of CHF, particularly under conditions when it is associated with CKD.

Background: Association of congestive heart failure (CHF) and chronic kidney disease (CKD) worsens the patient’s prognosis and results in poor survival rate. The aim of this study was to examine if addition of endothelin type A (ETA) receptor antagonist to the angiotensin-converting enzyme inhibitor (ACEi) will bring additional beneficial effects in experimental rats. Methods: CKD was induced by 5/6 renal mass reduction (5/6 NX) and CHF was elicited by volume overload achieved by creation of aorto-caval fistula (ACF). The follow-up was 24 weeks after the first intervention (5/6 NX). The treatment regimens were initiated 6 weeks after 5/6 NX and 2 weeks after ACF creation. Results: The final survival in untreated group was 15%. The treatment with ETA receptor antagonist alone or ACEi alone and the combined treatment improved the survival rate to 64%, 71% and 75%, respectively, however, the difference between the combination and either single treatment regimen was not significant. The combined treatment exerted best renoprotection, causing additional reduction in albuminuria and reducing renal glomerular and tubulointerstitial injury as compared with ACE inhibition alone. Conclusions: Our results show that treatment with ETA receptor antagonist attenuates the CKD- and CHF-related mortality, and addition of ETA receptor antagonist to the standard blockade of RAS by ACEi exhibits additional renoprotective actions.

Abstract Aims A retrospective nationwide observational analysis of diagnoses, procedures, and treatment reported to the Czech National Registry of Reimbursed Health Services between 2012 and 2018. Methods and results Prevalence of heart failure (HF) patients increased from 176 496 (1679.4 per 100 000 population) in 2012 to 285 745 (2689.0 per 100 000 population) patients in 2018 (mean age 74.4 ± 12.8 years). In the last years, a stable incidence of HF patients was observed (544 per 100 000 population in 2016 vs. 551 per 100 000 population in 2018; P = 0.310). Mortality rate decreased from 20.55% in 2012 to 15.89% in 2018. The number of hospitalized patients remained similar (318.2 per 100 000 population in 2012 vs. 311.8 per 100 000 population in 2018; P = 0.479). The most used drugs were diuretics (173 295; 60.6%) and beta‐blockers (178 823; 62.6%), followed by angiotensin‐converting enzyme inhibitors/angiotensin II receptor blockers (angiotensin‐converting enzyme inhibitors 120.581; 42.2%; angiotensin II receptor blockers 47 216; 16.5%). Even though the whole number of implanted devices in HF patients increased steadily (from 25 205 in 2012 to 45 363 in 2018), the prevalence of all devices (pacemakers and defibrillators) in the HF patients remained about the same (14.3% in 2012; 15.9% in 2018). Conclusions The study included all patients with HF in the Czech Republic. These are the first nationwide data of HF epidemiology in the Eastern bloc. The incidence of HF remains stable in the last years. Due to aging of the population, the prevalence of HF significantly increased in the last 6 years. Despite a continuous increase in the prevalence of HF and a suboptimal utilization of its pharmacological therapy, mortality decreased, and the number of hospitalized patients remained the same.

We showed recently that increasing kidney epoxyeicosatrienoic acids (EETs) by blocking soluble epoxide hydrolase (sEH), an enzyme responsible for EETs degradation, retarded the development of renal dysfunction and progression of aorto-caval fistula(ACF)-induced congestive heart failure (CHF) in Ren-2 transgenic hypertensive rats (TGR). In that study the final survival rate of untreated ACF TGR was only 14 % but increased to 41 % after sEH blockade. Here we examined if sEH inhibition added to renin-angiotensin system (RAS) blockade would further enhance protection against ACF-induced CHF in TGR. The treatment regimens were started one week after ACF creation and the follow-up period was 50 weeks. RAS was blocked using angiotensin-converting enzyme inhibitor (ACEi, trandolapril, 6 mg/l) and sEH with an sEH inhibitor (sEHi, c-AUCB, 3 mg/l). Renal hemodynamics and excretory function were determined two weeks post-ACF, just before the onset of decompensated phase of CHF. 29 weeks post-ACF no untreated animal survived. ACEi treatment greatly improved the survival rate, to 84 % at the end of study. Surprisingly, combined treatment with ACEi and sEHi worsened the rate (53 %). Untreated ACF TGR exhibited marked impairment of renal function and the treatment with ACEi alone or combined with sEH inhibition did not prevent it. In conclusion, addition of sEHi to ACEi treatment does not provide better protection against CHF progression and does not increase the survival rate in ACF TGR: indeed, the rate decreases significantly. Thus, combined treatment with sEHi and ACEi is not a promising approach to further attenuate renal dysfunction and retard progression of CHF.

We collected a multi-centric retrospective dataset of patients (N = 213) who were admitted to ten hospitals in Czech Republic and tested positive for SARS-CoV-2 during the early phases of the pandemic in March-October 2020. The dataset contains baseline patient characteristics, breathing support required, pharmacological treatment received and multiple markers on daily resolution. Patients in the dataset were treated with hydroxychloroquine (N = 108), azithromycin (N = 72), favipiravir (N = 9), convalescent plasma (N = 7), dexamethasone (N = 4) and remdesivir (N = 3), often in combination. To explore association between treatments and patient outcomes we performed multiverse analysis, observing how the conclusions change between defensible choices of statistical model, predictors included in the model and other analytical degrees of freedom. Weak evidence to constrain the potential efficacy of azithromycin and favipiravir can be extracted from the data. Additionally, we performed external validation of several proposed prognostic models for Covid-19 severity showing that they mostly perform unsatisfactorily on our dataset.

By omitting materials subject to the labeling requirement in part 1 cold-box resin, Düker GmbH from Laufach can achieve an impressive reduction in emissions.Emission and workplace exposure limit values represent major challenges for foundries, and are becoming increasingly important due to every stricter statutory regulations. In particular, compliance with emission guidelines is essential in safeguarding company sites, and is now of equal significance as technical and economic issues. Modern foundries such as Düker GmbH from Laufachpursue a sustainable corporate policy in which the three pillars of cost-effectiveness, social responsibility and environmental protection are accorded equal importance. Düker’s example shows how the use of the new ECOCURE BLUE technology supports environmental protection and occupational health and safety, while also facilitating efficient, highly productive core production.

Berberine, the most known quaternary protoberberine alkaloid (QPA), has been reported to inhibit the SIK3 protein connected with breast cancer. Berberine also appears to reduce the bcl-2 and XIAP expression-proteins responsible for the inhibition of apoptosis. As some problems in the therapy with berberine arose, we studied the DNA binding properties of escholidine, another QPA alkaloid. CD, fluorescence, and NMR examined models of i-motif and G-quadruplex sequences present in the n-myc gene and the c-kit gene. We provide evidence that escholidine does not induce stabilization of the i-motif sequences, while the interaction with G-quadruplex structures appears to be more significant.

This study evaluates the effects of chronic treatment with EET-A, an orally active epoxyeicosatrienoic acid (EETs) analog, on the course of aorto-caval fistula (ACF)-induced heart failure (HF) in Ren-2 transgenic rats (TGR), a model characterized by hypertension and augmented activity of the renin-angiotensin system (RAS). The results were compared with standard pharmacological blockade of the RAS using angiotensin-converting enzyme inhibitor (ACEi). The rationale for employing EET-A as a new treatment approach is based on our findings that apart from increased RAS activity, untreated ACF TGR also shows kidney and left ventricle (LV) tissue deficiency of EETs. Untreated ACF TGR began to die 17 days after creating ACF and were all dead by day 84. The treatment with EET-A alone or ACEi alone improved the survival rate: in 156 days after ACF creation, it was 45.5% and 59.4%, respectively. The combined treatment with EET-A and ACEi appeared to improve the final survival to 71%; however, the difference from either single treatment regimen did not reach significance. Nevertheless, our findings support the notion that targeting the cytochrome P-450-dependent epoxygenase pathway of arachidonic acid metabolism should be considered for the treatment of HF.

Apple trees (Malus domestica Borgh) are a rich source of dihydrochalcones, phenolic acids and flavonoids. Considering the increasing demand for these phytochemicals with health-benefitting properties, the objective of this study was to evaluate the profile of the main bioactive compounds—phloridzin, phloretin, chlorogenic acid and rutin—in apple tree bark, leaves, flower buds and twigs. The variety in the phenolic profiles of four apple tree cultivars was monitored during the vegetation period from March to September using chromatography analysis. Phloridzin, the major glycoside of interest, reached the highest values in the bark of all the tested cultivars in May (up to 91.7 ± 4.4 mg g−1 of the dried weight (DW), cv. ‘Opal’). In the leaves, the highest levels of phloridzin were found in cv. ‘Opal’ in May (82.5 ± 22.0 mg g−1 of DW); in twigs, the highest levels were found in cv. ‘Rozela’ in September (52.4 ± 12.1 mg g−1 of DW). In the flower buds, the content of phloridzin was similar to that in the twigs. Aglycone phloretin was found only in the leaves in relatively low concentrations (max. value 2.8 ± 1.4 mg g−1 of DW). The highest values of rutin were found in the leaves of all the tested cultivars (10.5 ± 2.9 mg g−1 of DW, cv. ‘Opal’ in September); the concentrations in the bark and twigs were much lower. The highest content of chlorogenic acid was found in flower buds (3.3 ± 1.0 mg g−1 of DW, cv. ‘Rozela’). Whole apple fruits harvested in September were rich in chlorogenic acid and phloridzin. The statistical evaluation by Scheffe’s test confirmed the significant difference of cv. ‘Rozela’ from the other tested cultivars. In conclusion, apple tree bark, twigs, and leaves were found to be important renewable resources of bioactive phenolics, especially phloridzin and rutin. The simple availability of waste plant material can therefore be used as a rich source of phenolic compounds for cosmetics, nutraceuticals, and food supplement preparation.

Objectives: Suboptimal medication adherence is common among patients with hypertension. Measurements of plasma or urinary levels of antihypertensive drugs are useful, but not widely available. The aim of our study was to investigate the relation of patients’ heart rates to their serum beta-blocker levels. Methods: We correlated 220 measurements of serum beta-blocker levels in 106 patients with apparently resistant hypertension to their corresponding office heart rate. A significant proportion, 44.6% of patients, were non-adherent to beta-blocker treatment according to serum level measurement. Non-adherent patients had significantly higher heart rates (80.9 vs. 66.6 bpm, p

An association between congestive heart failure (CHF) and chronic kidney disease (CKD) results in extremely poor patient survival rates. Previous studies have shown that increasing kidney epoxyeicosatrienoic acids (EETs) by blocking soluble epoxide hydrolase (sEH), an enzyme responsible for EETs degradation, improves the survival rate in CHF induced by aorto-caval fistula (ACF) and attenuates CKD progression. This prompted us to examine if sEH inhibitor treatment would improve the outcome if both experimental conditions are combined. Fawn-hooded hypertensive (FHH) rats, a genetic model showing early CKD development was employed, and CHF was induced by ACF. Treatment with an sEH inhibitor was initiated 4 weeks after ACF creation, in FHH and in fawn-hooded low-pressure (FHL) rats, a control strain without renal damage. The follow-up period was 20 weeks. We found that ACF FHH rats exhibited substantially lower survival rates (all the animals died by week 14) as compared with the 64% survival rate observed in ACF FHL rats. The former group showed pronounced albuminuria (almost 30-fold higher than in FHL) and reduced intrarenal EET concentrations. The sEH inhibitor treatment improved survival rate and distinctly reduced increases in albuminuria in ACF FHH and in ACF FHL rats, however, all the beneficial actions were more pronounced in the hypertensive strain. These data indicate that pharmacological blockade of sEH could be a novel therapeutic approach for the treatment of CHF, particularly under conditions when it is associated with CKD.

Aim To verify and compare the accuracies of mortality predictions in the Intensive Care Unit (ICU) of the Internal Clinic of Central Military Hospital in Prague, Czech Republic, using model APACHE II and the newer systems of the APACHE IV, SAPS 3 and MPMoIII. Methods The data were collected retrospectively between 2011 and 2012, 1000 patients were evaluated. The assessment of the overall accuracy of the mortality predictions was performed using the standardized mortality ratio (SMR), and the calibration was assessed using the Lemeshow-Hosmer “goodness-of-fit” C statistic. Discrimination was evaluated using ROC curves based on calculations of the areas under the curve (AUCs). Results The APACHE II, SAPS 3, and MPMoIII systems significantly overestimated the expected mortality, whereas the APACHE IV model led to correct estimations of the overall mortality. The discrimination capabilities of the models assessed according to the constructions of the ROC curves were evaluated as good, only the APACHE II was evaluated as satisfactory. The calibrations of all models were evaluated as unsatisfactory. Conclusion The best mortality estimation for the investigated population sample was provided by the APACHE IV system. The discrimination capabilities of all models for the studied population were satisfactory, but the calibration of all of the systems was unsatisfactory. The conclusions of our study are limited by the relatively small size of the investigated sample and the fact that this study was conducted at only a single site.

This paper attempts to show how studying the history of consumer culture can help to reveal identity-forming processes in Czech society. The author first outlines how this topic was highlighted in the German environment by the wave of ‘ostalgia’ to encourage, unlike the Czech milieu, systematic research into the everyday life under a Communist dictatorship. Subsequently, this paper briefly introduces the differences between Communist and Capitalist consumer culture. In the third part of the text, the author offers a possible extension of the research identity-forming processes exploring the development of the distribution infrastructure, the concepts of lifestyle and consumer discursive structures, advertising images of the national community and ultimately, monitoring the production of symbolic meanings of nation-specific objects.

Biofuels have become an integral part of everyday life in modern society. Bioethanol and fatty acid methyl esters are a common part of both the production of gasoline and diesel fuels. Also, pressure on replacing fossil fuels with bio-components is constantly growing. Waste vegetable fats can replace biodiesel. Hydrotreated vegetable oil (HVO) seems to be a better alternative. This fuel has a higher oxidation stability for storage purposes, a lower temperature of loss of filterability for the winter time, a lower boiling point for cold starts, and more. Viscosity, density, cold filter plugging point of fuel blend, and flash point have been measured to confirm that a fuel from HVO is so close to a fuel standard that it is possible to use it in engines without modification. The objective of this article is to show the properties of different fuels with and without HVO admixtures and to prove the suitability of using HVO compared to FAME. HVO can also be prepared from waste materials, and no major modifications of existing refinery facilities are required. No technology in either investment or engine adaptation of fuel oils is needed in fuel processing.

Mezi 28. a 30. zářím roku 2015 proběhl v Bruselu již 30. ročník setkání evropských studentů a zároveň se uskutečnila druhá PASCL konference. Tématem byla role vzdělávání s důrazem na perspektivu inkluze skrze přístup učení orientovaného na studenta. Konference probíhala v atraktivních prostorách Evropského parlamentu a Vlámské galerie vědy a umění.

In the growing seasons from 2003 to 2008, 547 isolates of Phytophthora infestans from five regions in the Czech Republic were collected and examined for their sensitivity to the active ingredients (metalaxyl, dimethomorph and propamocarb-HCl) of frequently used fungicides. The response of the isolates to each of these substances was examined using the in vitro amended-agar method; in 352 of these isolates, the sensitivity to metalaxyl was also assessed by the floating leaf-disc assay. The majority of the isolates were sensitive (89.8%) to metalaxyl. Resistant isolates were found only in two of the sample years (2003 and 2008); they represented 58% of the samples in 2003 and only 29% in 2008. Four isolates from 2004 were found to be intermediate for their level of resistance. All the isolates that were tested were sensitive to dimethomorph and propamocarb-HCl; these particular substances completely suppressed mycelial growth at 1 µg a.i. per ml.

A total of 195 isolates of Phytophthora infestans were collected from late blight-diseased potatoes grown in several localities in the Czech Republic during the years 2007-2008. The isolates were analysed for mating type using the pairing test, CAPS marker assays and PCR assays. Of the 195 tested isolates, 28% were of the A1 mating type and 75% corresponded to the A2 mating type. Furthermore, oospores of P. infestans were microscopically detected in leaf samples from one locality.

Quaternary benzo[c]phenanthridine alkaloids are secondary metabolites of the plant families Papaveraceae, Rutaceae, and Ranunculaceae with anti-inflammatory, antifungal, antimicrobial and anticancer activities. Their spectral changes induced by the environment could be used to understand their interaction with biomolecules as well as for analytical purposes. Spectral shifts, quantum yield and changes in lifetime are presented for the free form of alkaloids in solvents of different polarity and for alkaloids bound to DNA. Quantum yields range from 0.098 to 0.345 for the alkanolamine form and are below 0.033 for the iminium form. Rise of fluorescence lifetimes (from 2-5 ns to 3-10 ns) and fluorescence intensity are observed after binding of the iminium form to the DNA for most studied alkaloids. The alkanolamine form does not bind to DNA. Acid-base equilibrium constant of macarpine is determined to be 8.2-8.3. Macarpine is found to have the highest increase of fluorescence upon DNA binding, even under unfavourable pH conditions. This is probably a result of its unique methoxy substitution at C12 a characteristic not shared with other studied alkaloids. Association constant for macarpine-DNA interaction is 700000 M(-1).

A total of 199 Phytophthora infestans isolates were obtained from leaves, tubers and fruits of infected crops of potato and tomato in different regions of the Czech Republic in 2003, 2004 and 2005. They were analysed for mating type using the conventional pairing assay and PCR markers; 107 isolates were of A1 and 92 of A2 mating type. No self-fertile isolate was found. Our study is the first report of the presence and distribution of the A2 mating type of P. infestans in the Czech Republic. The co-existence of the two mating types may enable the pathogen to reproduce sexually, thus enhancing the diversity of its population countrywide.

The acute and chronic toxicity to honeybees, Apis mellifera of water guttation and dew collected from winter rape plants treated with the insecticide Nurelle D® (a.i. chlorpyriphos + cypermethrin) was investigated. Caged bees were fed on sugar syrup containing water guttation and dew for 24 h (acute toxicity test) and for 10 days (chronic toxicity test). Bee mortality and food consumption were determined daily. A contact toxicity test was performed within 24 h on bees kept in Petri dishes (10 bees per dish) lined with filter paper saturated with the test solution. The acute Nurelle D® contact and oral toxicity tests showed that the mortality of bees treated with water guttation and dew collected from the treated plants did not exceed 10%. A chronic toxicity test showed that adding contaminated water guttation and dew to the syrup caused an insignificant increase in bee mortality and reduced the syrup consumption significantly. The chlorpyriphos residue found in contaminated water guttation and dew were below the limit of detection (0.8 µg/kg) and the cypermethrin residue was below the detection levels.

The effects of NeemAzalTM formulations: NeemAzalTM T/S (1% azadirachtin) and NeemAzalTM granules (1% azadirachtin) on honeybees, Apis mellifera L., were studied under semi-field conditions. Three plots at 15 m2 each were sown with spring rape seeds Brassica napus cultivar Likolly (Brassicaceae/Cruciferae). In the first plot NeemAzal granules were added with the seeds during sowing. The second plot was sprayed with NeemAzal T/S during full flowering; GreemaxTM was used as a wetting agent. The third one was sprayed with water only during full flowering as a control. For each treatment one tunnel tent (3 × 5 × 2 m) was used during the flowering period. Small bee colonies were exposed to the treated plants for 7 days. Evaluation was carried out by comparing the results in the treatments to the control and, furthermore, by comparing the pre- and post-application. The mortality in the tunnels and the flight activity were checked before, as well as after the treatment. The development of the bee brood was evaluated by using transparent acetate sheets to mark single cells in brood combs with their contents on different assessment dates. The time schedule of the assessment dates was chosen in order to check the bee brood at different expected stages during the development. The development of the bee brood was evaluated by calculation of brood termination rates in percentage and brood indices. The results show that residues of NeemAzal granules did not adversely affect bee mortality, foraging activity or brood development. By contrast, it was noticed that NeemAzal T/S caused some reduction in foraging activity and brood development.

In this review, the latest information is given about the reference materials for the determination of contaminants in milk and milk products. Certified reference materials for the determination of contaminating chemical elements, organochlorine pesticides, polychlorinated biphenyls, dibenzodioxins, dibenzofurans, and aflatoxin M1 are presently available. Properties of these reference materials and possibilities for their use are discussed together with the maximum tolerances of contaminants in milk and milk products. The basic information about the producers and distributors of the reference materials in this area is provided.

Pathogens with rapid reproductive cycles, producing large number wind-spread dispersed infection propagules like sporangiospores, are more likely to pose problems of resistance than slowly reproducing pathogens, which are dispersed over only short distances. Phytophthora infestans, is highly adaptable pathogen capable of exerting high disease pressure on crop, and develop resistance to phenylamid fungicide methalaxyl in the potato crop in Europe (1981-1984). Metalaxyl was then reintroduced for use in mixtures coformulated with multisite compounds, and the proportion of resistance strains "subpopulations" rose again, and albeit to a level lower than at the outset. The use of metalaxyl on seed crops, with the possibility of resistance being carried over the next season. Current recommendations are to use alternative multiside fungicides on potato crops. Our results illustrate the importance of understanding pathogen epidemiology in the management of fungicide resistance.

The results of spatial variability of plant-available soil nutrients (P, K, Mg) and soil pH are described in this paper. Experiment was realized on the field of area 72 ha (orthic luvisol), located in the area of Český Brod. The use of coefficient of variation as a criterion of variability of soil agrochemical properties and yield on the field showed the following: the highest variability was observed in available P, the second highest variability was in available K, and the lowest variability of main non-mobile nutrients was in the available Mg. Soil pH was the lowest of all measured soil properties. Although the highest correlation coefficient between the soil available P content and soil pH was established, the process of spatial dependence was not detected. Detailed field scouting and others data can be important elements, as can complex decision rules, taking into account additional factors such as the characteristics of crop protection agents and preferences of the farm manager. This paper illustrates, how to plant nutritions, crop protection, crop production might be integrated to support these diseases and weeds management decisions.