Search

Copyright of Czecho-Slovak Pediatrics / Česko-Slovenská Pediatrie is the property of Czech Medical Association of JE Purkyne and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The PDCD1-encoded immune checkpoint receptor PD-1 is a key tumor suppressor in T cells that is recurrently inactivated in T cell non-Hodgkin lymphomas (T-NHLs). The highest frequencies of PDCD1 deletions are detected in advanced disease, predicting inferior prognosis. However, the tumor-suppressive mechanisms of PD-1 signaling remain unknown. Here, using tractable mouse models for T-NHL and primary patient samples, we demonstrate that PD-1 signaling suppresses T cell malignancy by restricting glycolytic energy and acetyl coenzyme A (CoA) production. In addition, PD-1 inactivation enforces ATP citrate lyase (ACLY) activity, which generates extramitochondrial acetyl-CoA for histone acetylation to enable hyperactivity of activating protein 1 (AP-1) transcription factors. Conversely, pharmacological ACLY inhibition impedes aberrant AP-1 signaling in PD-1-deficient T-NHLs and is toxic to these cancers. Our data uncover genotype-specific vulnerabilities in PDCD1-mutated T-NHL and identify PD-1 as regulator of AP-1 activity., (© 2023. The Author(s).)

In contrast to mono- or biallelic loss of tumor-suppressor function, effects of discrete gene dysregulations, as caused by non-coding (epi)genome alterations, are poorly understood. Here, by perturbing the regulatory genome in mice, we uncover pervasive roles of subtle gene expression variation in cancer evolution. Genome-wide screens characterizing 1,450 tumors revealed that such quasi-insufficiency is extensive across entities and displays diverse context dependencies, such as distinct cell-of-origin associations in T-ALL subtypes. We compile catalogs of non-coding regions linked to quasi-insufficiency, show their enrichment with human cancer risk variants, and provide functional insights by engineering regulatory alterations in mice. As such, kilo-/megabase deletions in a Bcl11b -linked non-coding region triggered aggressive malignancies, with allele-specific tumor spectra reflecting gradual gene dysregulations through modular and cell-type-specific enhancer activities. Our study constitutes a first survey toward a systems-level understanding of quasi-insufficiency in cancer and gives multifaceted insights into tumor evolution and the tissue-specific effects of non-coding mutations., Competing Interests: The authors declare no competing interests., (© 2023 The Authors.)

SNAIL is a key transcriptional regulator in embryonic development and cancer. Its effects in physiology and disease are believed to be linked to its role as a master regulator of epithelial-to-mesenchymal transition (EMT). Here, we report EMT-independent oncogenic SNAIL functions in cancer. Using genetic models, we systematically interrogated SNAIL effects in various oncogenic backgrounds and tissue types. SNAIL-related phenotypes displayed remarkable tissue- and genetic context-dependencies, ranging from protective effects as observed in KRAS- or WNT-driven intestinal cancers, to dramatic acceleration of tumorigenesis, as shown in KRAS-induced pancreatic cancer. Unexpectedly, SNAIL-driven oncogenesis was not associated with E-cadherin downregulation or induction of an overt EMT program. Instead, we show that SNAIL induces bypass of senescence and cell cycle progression through p16 INK4A -independent inactivation of the Retinoblastoma (RB)-restriction checkpoint. Collectively, our work identifies non-canonical EMT-independent functions of SNAIL and unravel its complex context-dependent role in cancer., (© 2023. The Author(s).)

Food insecurity and perinatal depression are significant public health concerns for perinatal services, however descriptive research examining their association is limited. The purpose of this study was to examine the views and perspectives of staff from the Special Supplemental Nutrition Program for Women, Infants and Children (WIC) program on the relationship between food insecurity and perinatal depression among their WIC clients. Four, semi-structured focus groups with WIC staff ( n = 24) were conducted across four diverse nonmetropolitan public health districts in Midwestern counties in the United States. WIC staff included social workers, nurses, nutritionists and ancillary staff. All interviews were audio-recorded, transcribed, and verified, and data were organized using NVivo 11.4.2. Thematic networking analysis was employed as the qualitative analysis to identify organizing themes. Three themes emerged including (1) depression experienced by clients; (2) food insecurity experienced by clients; and (3) barriers preventing clients from accessing services for themselves and their children. Research on food insecurity and perinatal depression is sparse, with fewer studies having included health staff of low-income women. Our findings suggest that the association between food insecurity and mental health needs among WIC clients is a significant public health issue to which policy change and interventions are required.

Enteroviruses (EVs) are associated with a wide spectrum of diseases involving various organs. Our aim was to give a historical overview of the genesis of clinical sample processing for EVs in the Slovak Republic (SR) during the 1958-2020 period, within the framework of the World Health Organization (WHO) polio program. Further, analyses were made of the data obtained from the archives of processed clinical sample surveillance using statistical methods. We used generalized additive models (GAM) with binomial distribution and logit link functions and an autoregressive moving average (ARMA) to analyze the data obtained during this 63-year period. Our results show trends in the composition of EV strains circulating in the population. Furthermore, statistically significant increasing trends of the non-polio enteroviruses (NPEVs) were observed over the studied time, represented by echoviruses (E) and coxsackieviruses A and B (CVA and CVB), with a cyclical pattern of occurrence. The most prevalent serotype over this period was CVB5, which became significantly more prevalent after 2000. While PVs, CVB1, and CVB3 were present in the second half of the studied period, CVA10, CVA16, E3, E25, and E30 appeared more frequently.

We describe the genesis of poliovirus (PV) and non-polio enterovirus (NPEV) surveillance program of sewage wastewaters from its inception to the present in the Slovak Republic (SR). Sampling procedures and evolution of the methodology used in the SR for the detection of PVs and NPEVs are presented chronologically. For statistical data processing, we divided our dataset into two periods, the first period from 1963 to 1998 (35 years), and the second period from 1999 to 2019 (21 years). Generalized additive models were used to assess temporal trends in the probability of occurrence of major EV serotypes during both periods. Canonical correspondence analysis on relative abundance data was used to test temporal changes in the composition of virus assemblages over the second period. The probability of occurrence of major viruses PV, coxsackieviruses (CVA, CVB), and Echoviruses (E)) significantly changed over time. We found that 1015 isolated PVs were of vaccine origin, called "Sabin-like" (isolates PV1, PV2, PV3). The composition of EV assemblages changed significantly during the second period. We conclude that during the whole period, CVB5, CVB4, and E3 were prominent NPEVS in the SR.

The mortality of patients with pancreatic ductal adenocarcinoma (PDAC) is strongly associated with metastasis, a multistep process that is incompletely understood in this disease. Although genetic drivers of PDAC metastasis have not been defined, transcriptional and epigenetic rewiring can contribute to the metastatic process. The epigenetic eraser histone deacetylase 2 (HDAC2) has been connected to less differentiated PDAC, but the function of HDAC2 in PDAC has not been comprehensively evaluated. Using genetically defined models, we show that HDAC2 is a cellular fitness factor that controls cell cycle in vitro and metastasis in vivo, particularly in undifferentiated, mesenchymal PDAC cells. Unbiased expression profiling detected a core set of HDAC2-regulated genes. HDAC2 controlled expression of several prosurvival receptor tyrosine kinases connected to mesenchymal PDAC, including PDGFRα, PDGFRβ, and EGFR. The HDAC2-maintained program disabled the tumor-suppressive arm of the TGFβ pathway, explaining impaired metastasis formation of HDAC2-deficient PDAC. These data identify HDAC2 as a tractable player in the PDAC metastatic cascade. The complexity of the function of epigenetic regulators like HDAC2 implicates that an increased understanding of these proteins is needed for implementation of effective epigenetic therapies., Significance: HDAC2 has a context-specific role in undifferentiated PDAC and the capacity to disseminate systemically, implicating HDAC2 as targetable protein to prevent metastasis., (©2021 The Authors; Published by the American Association for Cancer Research.)

Non-pharmaceutical interventions (NPIs) were the only viable choice to mitigate or suppress transmission of COVID-19 in the absence of efficient and safe vaccines. In this study, we examined the association between the stringency of containment measures and cumulative incidence of the COVID-19 cases in the first wave of the pandemic across 28 European countries. Our results support the effectiveness of containment measures in the mitigation or suppression of COVID-19 epidemics. Early adoption of stringent containment measures prior to detection of the first confirmed case, together with ramping up containment stringency during the early days of epidemics, was associated with a lower disease occurrence. The delayed adoption of stringent containment measures did not fully compensate for the lack of early response. Containment measures continue to play a significant role in the control of COVID-19 in the post-vaccination period, when limited vaccination coverage, the emergence of vaccine resistance, and/or increased mobility enabled further disease transmission (Tab. 4, Fig. 22, Ref. 50). Keywords: non-pharmaceutical interventions, containment, COVID-19, mobility, social distancing, Containment and Health Index, epidemiology, public health measures, SARS-CoV-2.

Numerous serotypes which belong to the genus Enterovirus (EV) show variability in their virulence and clinical manifestations. They are also known to undergo changes caused by mutations and recombination during their circulation in the environment and the population. Various EV serotypes are prevalent in groundwater, wastewater and surface waters. Our previous studies showed that oral infection induces pancreatitis depending on specific conditions, such as gravidity, in an outbred murine model. Our aim in the present study was to further explore the pancreatic histopathology in an outbred mouse model following oral infection with clinical isolates from a patient who had aseptic meningitis and an isolate from a treated-sewage sample recovered from the residential area of the patient. The isolates were identified as coxsackievirus B4 (CVB4) in tissue culture. The CVB4 sewage-isolate induced pancreatitis after oral infection. In contrast, pancreatitis was absent following infection with the clinical isolates. Comparison of polyprotein sequences showed that the treated-sewage strains differed from the patient's isolates by 9 and 11 amino acids. We conclude that the isolates of clinical and environmental origin differed in their pathogenic properties and showed genetic variation., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.)

In times of COVID-19 pandemics, the upcoming period of the year when influenza activity usually increases in the Northern Hemisphere brings new medical and public health challenges. These challenges include the risk of mixed infections and/or a possible collision of the two epidemics (“twindemia”) with a potentially serious impact on individual health and public health. In this report, we discuss the results of the published stu-dies and conclude that the catastrophic collision of the seasonal influenza and COVID-19 epidemics is unlikely when efficient non-pharmaceutical public health measures are applied to control or mitigate the spread of the COVID-19 epidemic. This conclusion is supported by several lines of evidence, including the extremely low seasonal influenza activity registered in the Southern Hemisphere in 2020. On the other hand, the existence of mixed SARS-CoV-2 and influenza virus infections has been demonstrated in humans. The continuing uncertainty about the occurrence and potential severity of these mixed infections emphasizes the importance of seasonal influenza vaccination in the current epidemiological situation and raises the need to: (i) ensure vaccine availability, (ii) facilitate access to safe seasonal influenza vaccination under the conditions of the ongoing COVID-19 epidemic, and (iii) promote the vaccine to the public.

Objective: In most indicators of the way of life, the Roma community is generally different from the majority population and dominant culture. The objective of the study was to describe factors affecting the health of the Roma living in Slovakia, with an emphasis on the sexual and reproductive health of Roma women, and report on the results of analysis of high-risk pregnancies of Roma women in the district of Rimavská Sobota, Slovakia., Methods: A retrospective study of medical documentation was used. The results were analyzed using the absolute and relative frequencies. Statistical methods were used., Results: A total of 1,256 high-risk pregnancies were analyzed, of which 622 (49.52%) were in Roma women. The average age of Roma respondents was lower by 5 years compared to non-Roma. The age of Roma women at the first pregnancy was statistically significantly lower compared to non-Roma (p < 0.001). The Roma respondents achieved statistically significantly lower levels of education than non-Roma. There was a demonstrably higher number of pregnancies as well as a higher number of artificial and spontaneous abortions per Roma woman. These results were statistically significant. For Roma women, pregnancy began to be risky demonstrably earlier than for non-Roma (p < 0.001). There was a statistically significant difference in attending prenatal counselling. Roma women attended prenatal counselling statistically significantly less frequently than non-Roma (p < 0.001). A significant statistical dependence was found between attending prenatal counselling and the onset of pregnancy problems in Roma women. There was no significant difference in the incidence of other diseases associated with high-risk pregnancy among Roma and non-Roma respondents., Conclusion: The findings indicate that Roma women are exposed to health problems in the area of sexual and reproductive health in Slovakia. In the approach to the Roma, it is essential to focus on improving accessibility to health care, prevention, knowledgeableness and effectively preventing, eradicating and strongly penalizing all forms of discrimination in access to health care, especially for Roma women, who are more likely to receive health care.

The number of outlaw bikers is growing globally. Despite this, little research exists on these groups and their alleged violent tendencies. To address this, the current paper uses unique data to examine whether gang violence causes outlaw biker violence. The period examined runs from mid-2008 until early 2012 during which violent clashes occurred between outlaw bikers and street gang members involved in an alleged conflict in Copenhagen, Denmark. A precise description of each individual act of violence would make it possible to identify whether specific acts were carried out in furtherance of the alleged conflict. This would allow one to determine whether outlaw bikers commit violence on behalf of their club. However, such knowledge is unavailable. The paper therefore takes a different approach by examining whether acts of violence committed by the two groups are statistically associated. In other words, it considers whether one or more acts can be described as retaliatory during the observation periods. The sample consists of 640 individuals involved with the Hells Angels Motorcycle Club or with non-biker street gangs-both of which are present in Copenhagen. Statistical models are used to predict 143 violent events committed by 196 outlaw bikers. The results suggest that violence committed by gang members predicts violence committed by outlaw bikers. This indicates that violent acts committed by outlaw bikers are at least partly a form of retaliation carried out on behalf of their club. The paper expands the literature on the kinds of inter-group, micro-level processes that can lead to reciprocal violence by including outlaw bikers in a literature that has previously focused on non-biker street gangs., Competing Interests: The author has declared that no competing interests exist.

A review is presented of achievements of Czechoslovak, Czech, and Slovak microbiologists in the identification of microorganisms named after geographical locations or their discoverers. Without a fortunate coincidence of circumstances and the rigorous work of microbiologists, there could be gaps in our scientific knowledge we use to our benefit and which will always remain part of the heritage in the area of microbiology of both nations - Czechs and Slovaks. . KEYWORDS: microbiology - history - geography - Czechoslovak microbiologists - bacteria - viruses.

A case of food-borne botulism occurred in Slovakia in 2015. Clostridium botulinum type A was isolated from three nearly empty commercial hummus tubes. The product, which was sold in Slovakia and the Czech Republic, was withdrawn from the market and a warning was issued immediately through the European Commission's Rapid Alert System for Food and Feed (RASFF). Further investigation revealed the presence of botulinum neurotoxin (BoNT) subtype BoNT/A3, a very rare subtype implicated in only one previous outbreak (Loch Maree in Scotland, 1922). It is the most divergent subtype of BoNT/A with 15.4% difference at the amino acid level compared with the prototype BoNT/A1. This makes it more prone to evading immunological and PCR-based detection. It is recommended that testing laboratories are advised that this subtype has been associated with food-borne botulism for the second time since the first outbreak almost 100 years ago, and to validate their immunological or PCR-based methods against this divergent subtype., (This article is copyright of The Authors, 2017.)

The lymphoid-myeloid transdifferentiation potentials of members of the C/EBP family (C/EBPα, β, δ, and ε) were compared in v-Abl-immortalized primary B cells. Conversion of B cells to macrophages was readily induced by the ectopic expression of any C/EBP, and enhanced by endogenous C/EBPα and β activation. High transgene expression of C/EBPβ or C/EBPε, but not of C/EBPα or C/EBPδ, also induced the formation of granulocytes. Granulocytes and macrophages emerged in a mutually exclusive manner. C/EBPβ-expressing B cells produced granulocyte-macrophage progenitor (GMP)-like progenitors when subjected to selective pressure to eliminate lymphoid cells. The GMP-like progenitors remained self-renewing and cytokine-independent, and continuously produced macrophages and granulocytes. In addition to their suitability to study myelomonocytic lineage bifurcation, lineage-switched GMP-like progenitors could reflect the features of the lympho-myeloid lineage switch observed in leukemic progression., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

Human foodborne botulism is an intoxication caused by ingestion of botulinum neurotoxins (BoNT) of serotypes A, B, E, and rarely also serotype F, produced in contaminated food by anaerobic bacteria Clostridium botulinum group I, group II, or by toxigenic strains of C. butyricum and C. baratii. BoNT-producing Clostridia are ubiquitously distributed in the environment and, under suitable conditions, they can enter the food chain, proliferate and produce BoNT in a variety of foods. In the past, the risk of foodborne botulism was primarily associated with homemade canned foods; however, the epidemiological importance of commercial and restaurant food is increasing nowadays. In this article, we review the public health aspects of foodborne botulism, including its clinical, epidemiological and laboratory diagnosis and discuss potential risks associated with minimally heated, vacuum or modified atmosphere-packed, ready-to-eat foods of extended durability.

Research has exposed cancer to be a heterogeneous disease with a high degree of inter-tumoral and intra-tumoral variability. Individual tumors have unique profiles, and these molecular signatures make the use of traditional histology-based treatments problematic. The conventional diagnostic categories, while necessary for care, thwart the use of molecular information for treatment as molecular characteristics cross tissue types.This is compounded by the struggle to keep abreast the scientific advances made in all fields of science, and by the enormous challenge to organize, cross-reference, and apply molecular data for patient benefit. In order to supplement the site-specific, histology-driven diagnosis with genomic, proteomic and metabolomics information, a paradigm shift in diagnosis and treatment of patients is required.While most physicians are open and keen to use the emerging data for therapy, even those versed in molecular therapeutics are overwhelmed with the amount of available data. It is not surprising that even though The Human Genome Project was completed thirteen years ago, our patients have not benefited from the information. Physicians cannot, and should not be asked to process the gigabytes of genomic and proteomic information on their own in order to provide patients with safe therapies. The following consensus summary identifies the needed for practice changes, proposes potential solutions to the present crisis of informational overload, suggests ways of providing physicians with the tools necessary for interpreting patient specific molecular profiles, and facilitates the implementation of quantitative precision medicine. It also provides two case studies where this approach has been used., Competing Interests: There is no conflict of interest.

Sex differences in glucocorticoid production are associated with increased responsiveness of the adrenal gland in females. However, the adrenal-intrinsic mechanisms that establish sexual dimorphic function remain ill defined. Glucocorticoid production is gated at the molecular level by the circadian clock, which may contribute to sexual dimorphic adrenal function. Here we examine sex differences in the adrenal gland using an optical reporter of circadian clock function. Adrenal glands were cultured from male and female Period2::Luciferase (PER2::LUC) mice to assess clock function in vitro in real time. We confirm that there is a pronounced sex difference in the intrinsic capacity to sustain PER2::LUC rhythms in vitro, with higher amplitude rhythms in adrenal glands collected from males than from females. Changes in adrenal PER2::LUC rhythms over the reproductive life span implicate T as an important factor in driving sex differences in adrenal clock function. By directly manipulating hormone levels in adult mice in vivo, we demonstrate that T increases the amplitude of PER2::LUC rhythms in adrenal glands of both male and female mice. In contrast, we find little evidence that ovarian hormones modify adrenal clock function. Lastly, we find that T in vitro can increase the amplitude of PER2::LUC rhythms in male adrenals but not female adrenals, which suggests the existence of sex differences in the mechanisms of T action in vivo. Collectively these results reveal that activational effects of T alter circadian timekeeping in the adrenal gland, which may have implications for sex differences in stress reactivity and stress-related disorders.

Influenza is one of the most common human infectious diseases, and has profound health and economic consequences. The laboratory diag- nosis of influenza virus infections plays an important role in the global surveillance of influenza. Therefore, there is a growing demand for highly sensitive and rapid methods for detecting influenza. The performance of particular diagnostic methods is affected by various factors. In this study, we assess the effects of patients' age and time to diagnosis on the probability of detecting influenza using four diagnostic methods (virus isolation, rapid test, RT-PCR and real-time RT-PCR). We examined 3,546 samples from central and eastern Slovakia during the influenza seasons from 2005-2006 to 2010-2011. In general, the probability of influenza detection significantly decreased with the time from onset of illness to sample collection (T1) as well as with patients' age (AGE). On the contrary, time from sample collection to delivery (T2) did not play a role in the prob- ability of influenza detection. As judged by odds ratios, the virus isolation method was most sensitive to T1, followed by the rapid test and RT-PCR methods. For the effect of AGE, the rapid test and virus isolation methods were more sensitive than PCR-based methods. The effects of T1 and AGE were independent of each other. Laboratories which participate in inifluenza surveillance should use several methods to enable rapid and accurate influenza A and B virus detection.

Unlabelled: The Department of Medical Microbiology of the Regional Authority of Public Health (RAPH) in Banská Bystrica serves as a catchment laboratory of virology for the Central Slovakia Region, and in the influenza season 2009/10, it also served as such for the East Slovakia Region., Material and Methods: Specimens (nasopharyngeal swabs and post-mortem specimens) from patients with suspected influenza were obtained from both sentinel and non-sentinel physicians. The specimens were analyzed by a rapid test, followed by real-time PCR (RT-PCR) for influenza A or B diagnosis. RT-PCR subtyping for pandemic influenza A/H1N1 was performed., Results: From May 2009 to June 2010, 2497 specimens were analyzed for the presence of influenza A and B viruses and in particular for the presence of pandemic influenza A/H1N1 virus. As many as 537 of 589 influenza A-positive specimens, i.e. 21.5% of all specimens analyzed and 91.2% of influenza A-positive specimens, were subtyped as pandemic influenza A/H1N1., Conclusion: In the influenza season 2009/10, the new pandemic influenza A/H1N1 clearly predominated in Central and Eastern Slovakia. PCR tests have played a key role in diagnosing patients with suspected pandemic influenza in the laboratory participating in the surveillance of influenza and influenza-like illness in the Slovak Republic.

Naegleria fowleri is a free-living amoeba that can cause primary amoebic meningoencephalitis (PAM). While, traditional methods for diagnosing PAM still rely on culture, more current laboratory diagnoses exist based on conventional PCR methods; however, only a few real-time PCR processes have been described as yet. Here, we describe a real-time PCR-based diagnostic method using hybridization fluorescent labelled probes, with a LightCycler instrument and accompanying software (Roche), targeting the Naegleria fowleriMp2Cl5 gene sequence. Using this method, no cross reactivity with other tested epidemiologically relevant prokaryotic and eukaryotic organisms was found. The reaction detection limit was 1 copy of the Mp2Cl5 DNA sequence. This assay could become useful in the rapid laboratory diagnostic assessment of the presence or absence of Naegleria fowleri., (Copyright 2009 Elsevier Inc. All rights reserved.)

Background: The aim of the present study was to monitor the nasopharyngeal presence of Streptococcus pneumoniae in different age groups (especially children) in Banská Bystrica, Slovakia. The purpose of this screening was to determine the prevalence of different serotypes and to follow up the presence of pneumococcus in these children after the vaccination with heptavalent protein-conjugate vaccine. A contribution of molecular biology techniques was the detection of S. pneumoniae DNA by PCR and also the typisation and comparison of pneumococcal strains by pulsed-field gel electrophoresis., Methods: S. pneumoniae in nasopharyngeal swabs was detected by cultivation on blood agar plates. Serotypisation was performed by standard Quellung reaction. The commercial diagnostic kit was used for PCR detection of S. pneumoniae DNA. Pulsed-field electrophoresis was performed by modified scheme according to literature., Results: The incidence of pneumococcus is decreasing and less significant with the increasing age. Among youngest children is relatively high prevalence of pneumococci and the relatedness of isolated strains is high as well. After the vaccination, the less invasive serotypes were detected, although the overall incidence of S. pneumoniae was similar., Conclusions: The monitoring of S. pneumoniae in population is important according to variability of this bacteria with respect to possible changes in pneumococcal types as a consequence of vaccination.

Acanthamoeba keratitis is rare corneal disease, its etiology is caused by amoebae of the Acanthamoeba spp. In this paper, the newest findings about the diagnostic and treatment procedures of the disease and epidemiology and preventive issues from the point of public health are presented. The article presents results of the water quality monitoring (according to the Acanthamoeba which is possible to cultivate at 36 degrees C and 44 degrees C) in man-made swimming pools during the period 2004-2008 at the Department of environmental biology of the Regional Public Health Institute in Banská Bystrica, Slovakia, E.U. The examination methods present the techniques of the clinical sampling and taking samples from the environment. The results underline the use of the new effective criteria in controlling of recreational resorts as well as changes of the legal criteria for the water quality used by the public. The results show that the presence of Acanthamoeba spp. in the environment is common, so the water monitoring is perceived as substantial preventive issue to prevent the disease to emerge. The collaboration between the public heath departments and ophthalmologists during the examination of the clinical and environmental samples may help to prevent and diagnose the Acanthamoeba keratitis.

Under Central European climatic conditions, toxoplasmosis is one of the most common human parasitic diseases. A wide range of methods for both direct and indirect detection of the causative agent are currently available for the laboratory diagnosis of toxoplasmosis. The purpose of the article is to review the history of the discovery of the causative agent of toxoplasmosis and how laboratory diagnostic methods were developed and improved. The main emphasis is placed on current options in the diagnosis of Toxoplasma gondii, more precisely on the serodiagnosis and new trends in molecular biology-based techniques.

Many severe diseases of the respiratory tract lead to hospitalisation. These diseases are often caused by viral infections and may cause increased mortality. The most common viral pathogens involved in these cases, which are also associated with significant morbidity and mortality during the influenza seasons are influenza viruses. Rapid differential diagnosis of influenza viruses is therefore of great importance. Classical diagnosis of these viruses involves virus cultures. Of the rapid diagnostic methodologies which have been developed are RT-PCR, multiplex PCR, real-time PCR. In the present study we have monitored clinical samples from patients of different age groups from selected regions in Slovakia and compared the effectiveness of the classical and molecular biological diagnostic methods. The molecular biological methods proved to be rapid, accurate and effective. Application of these techniques in diagnosis of the respiratory illnesses should help in the prevention, therapy and disease control.

Pulsed-field gel electrophoresis was developed for separating and analyzing of long DNA fragments in alternating electric field. In homogenous electric field, fragments longer than 50 kb run as a broad, unresolved band with high mobility. PFGE separated the DNA by periodicaly changing the direction of electric field. DNA molecules are moving "zig-zag" through the gel and they can be better separated. Fragments of several megabases can be resolved using this method. PFGE can be used for genome mapping of microorganisms as well as higher organisms. In microbiology, PFGE is a standard method for typization of bacteria. Comparison of electrophoresis profiles after digestion od DNA from bacterial isolates with restriction endonuclease is a very useful epidemiologists tool. Genetically identical organisms have the same PFGE profiles, different strains have different profiles. Related strains have also similar electrophoretic profiles. This enables to determine if the outbreaks are caused by the same strain of microorganism, to locate the source of outbreak and to monitor the spread of the microorganism. The most followed-up are nosocomial and the food-borne pathogens. PFGE can be also used for monitoring genetic evolution of the microorganism and the most prevalent types which circulate in population. This can be very useful for preparation of vaccines.

Objective: To assess the prevalence of hepatitis C in the general Slovak population without any further evaluation or risk group stratification., Material and Methods: A total of 3,608 remnant serum specimens from epidemiological surveys in 1997 (1,484 specimens) and 2002 (2,124) were analyzed. These were from randomly selected persons over 15 years of age from all over Slovakia. The anti-HCV antibodies were detected using the 4th generation ELISA test. In case of positive or borderline results, the presence of HCV RNA was determined qualitatively., Results: Of the 3,608 analyzed specimens, 55 (1.52 %) were anti-HCV-positive and 10 (0.28 %) were borderline positive. HCV RNA was detected in 24 cases (0.67 %). A comparison of the 1997 and 2002 results showed a statistically significant (p < 0.01) increase of anti-HCV-positive specimens. A similar--but not significant--increase was noted in HCV RNA-positive cases. Despite a slightly higher prevalence of HCV infection in females, no statistically significant gender differences were found. Whereas anti-HCV positivity increased slightly with age, most HCV RNA patients were from the middle age group, i.e. between 36 and 45 years of age. The geographic distribution of HCV cases across Slovakia was relatively even., Conclusions: The prevalence of anti-HCV antibodies in subjects representing the general population of Slovakia older than 15 years was 1.52 %; chronic HCV infection was confirmed in 0.67 % of cases.

By application of the real-time PCR we manage to confirm the diagnosis and occurrence of a disease, which is caused by Bordetella pertussis-pertussis. Using this method we have proven the presence of DNA of Bordetella pertussis in the biological materials (nasopharyngeal swabs). The presence of IS481 genome sequence of Bordetella pertussis was confirmed. This method of detection of pathogens seems to be very rapid, simple, and specific. In the case of adequate technical laboratory equipment it may become very suitable and important supporter in explanation and confirmation of the occurrence of bacterial infections.