BACKGROUND: Antiviral resistance development is a serious complication of human cytomegalovirus virostatic therapy caused by mutations in UL 97 and/or UL54 genes. OBJECTIVES: To determinate the presence of sensitive and resistant strains in patients developing antiviral resistance. STUDY DESIGN: We used three different molecular biological methods for mutation analysis-restriction fragment length polymorphism, sequencing and real-time PCR approach. RESULTS: We describe three allogeneic hematopoietic stem cell transplant patients developing the GCV resistant HCMV strains manifested by virostatic treatment failure. In these patients we identified UL97 mutations L595S, A594V and A594T and monitored the dynamics of coexisted sensitive/resistant strains. We confirmed the presence of mixed HCMV populations and in two patients a phenomenon of sensitive strain repopulation which occurred after 6.5 months and 1 month after removing GCV pressure. CONCLUSIONS: Our results show changes in proportions of sensitive/resistant subpopulations over time but other studies would be required to demonstrate the beneficial impact of their monitoring on clinical outcome.

• Klíčová slova
• HCMV resistance, Real-time PCR, Repopulation, Sensitive/resistant strain,
• MeSH
• antivirové látky terapeutické užití   MeSH
• cytomegalovirové infekce farmakoterapie   virologie   MeSH
• Cytomegalovirus účinky léků   genetika   izolace a purifikace   MeSH
• diagnostické techniky molekulární metody   MeSH
• dospělí MeSH
• homologní transplantace škodlivé účinky   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• missense mutace * MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• sekvenční analýza DNA MeSH
• terapie neúspěšná MeSH
• transplantace hematopoetických kmenových buněk škodlivé účinky   MeSH
• virová léková rezistence * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antivirové látky MeSH

Forests account for nearly 90 % of the world's terrestrial biomass in the form of carbon and they support 80 % of the global biodiversity. To understand the underlying forest dynamics, we need a long-term but also relatively high-frequency, networked monitoring system, as traditionally used in meteorology or hydrology. While there are numerous existing forest monitoring sites, particularly in temperate regions, the resulting data streams are rarely connected and do not provide information promptly, which hampers real-time assessments of forest responses to extreme climate events. The technology to build a better global forest monitoring network now exists. This white paper addresses the key structural components needed to achieve a novel meta-network. We propose to complement - rather than replace or unify - the existing heterogeneous infrastructure with standardized, quality-assured linking methods and interacting data processing centers to create an integrated forest monitoring network. These automated (research topic-dependent) linking methods in atmosphere, biosphere, and pedosphere play a key role in scaling site-specific results and processing them in a timely manner. To ensure broad participation from existing monitoring sites and to establish new sites, these linking methods must be as informative, reliable, affordable, and maintainable as possible, and should be supplemented by near real-time remote sensing data. The proposed novel meta-network will enable the detection of emergent patterns that would not be visible from isolated analyses of individual sites. In addition, the near real-time availability of data will facilitate predictions of current forest conditions (nowcasts), which are urgently needed for research and decision making in the face of rapid climate change. We call for international and interdisciplinary efforts in this direction.

• Klíčová slova
• Automated, standardized linking methods, Ecophysiology, Forest monitoring and observation infrastructure, Meta-network, Nowcasting and predictions in near real-time, Remote sensing,
• Publikační typ
• časopisecké články MeSH

The recent availability of genome information greatly facilitates the fundamental research on chicken. In different organs, gene expression patterns can provide clues to understanding the biological functions. For rapid and accurate quantification of gene expression, quantitative real-time PCR (qPCR) has become one of the most widely used methods. However, the success of qPCR data normalization depends on the use of a suitable reference gene and a single reference gene is not universally suitable for all the experiments. Therefore, reference gene validation is a crucial step for different organ tissues of chicken where suitable reference genes for qPCR analysis in varieties of tissues have not been investigated exhaustively so far. In this study, we have selected 30 Gallus gallus candidate reference genes from NCBI, amplified and studied their expression profiles by qPCR in different organ tissues (breast muscle, thigh muscle, heart, liver, spleen, gizzard, and bursa) of chicken. The result showed that, for breast muscle HSP10 and RPL23, thigh muscle RPL14 and RPL13, liver ALB and HSP70, spleen ALB and GAPDH, heart CYCS and TUBA8B, gizzard RPL5 and 18S rRNA, and bursa EEF1A1 and PGK2 are most stable genes respectively. The results also showed that for different organ tissues, individual or a combination of reference genes should be selected for data normalization. In this study, we have identified and validated 30 reference genes in seven different organ tissues to provide accurate transcript normalization and quantification, which can be useful for gene expression studies in other avian species.

• Klíčová slova
• chicken, normalization, organ tissues, quantitative real-time PCR, reference gene,
• MeSH
• exprese genu MeSH
• kosterní svaly MeSH
• kur domácí * genetika   MeSH
• kvantitativní polymerázová řetězová reakce veterinární   MeSH
• referenční standardy MeSH
• stanovení celkové genové exprese * veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The aim of this work was to identify reliable reference genes for expression studies in adult Haemonchus contortus. Eleven candidate genes were identified and the stability of their expression was assessed in adult males and females of two genetically divergent H. contortus isolates: drug-susceptible (ISE) and multi-drug-resistant (WR). Five genes with the most stable expression patterns were further assessed for suitability as reference genes in anthelmintic-treated H. contortus adults versus non-treated controls. We identified important differences in the expression of a number of candidate genes in anthelmintic-treated samples, confirming the need for careful validation of control genes for such experiments. We propose the use of multiple reference genes for expression studies in this species and found gpd, ama and far most suitable for adult H. contortus.

• Klíčová slova
• Anthelmintic, Haemonchus contortus, Reference gene, qPCR,
• MeSH
• anthelmintika farmakologie   MeSH
• Haemonchus účinky léků   genetika   MeSH
• kvantitativní polymerázová řetězová reakce metody   normy   MeSH
• referenční standardy * MeSH
• stanovení celkové genové exprese metody   normy   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• anthelmintika MeSH

Water quality monitoring using fish and crayfish as bio-indicators requires an understanding of the state of pollution of waters, choice of bio-indicators, physiological and behavioral endpoints of fish and crayfish, and principles of the methodology and their potential applications. Here, we discuss telemetry, acoustic monitoring, vision-based monitoring, measures of ventilatory activity, electrocardiography, and fiber-optic plethysmography. Assessment of water quality must be based, not only on physicochemical characteristics of the current environment as determined by chemical analyses, but also on observations of the physiology and behavior of its inhabitants. Real-time biomonitoring is suggested as the most reliable method, since it incorporates living organisms into the system to serve as biosensors. The potential application of the methods discussed includes use at water treatment plants and water supply stations for prevention of hazardous toxicological events, and, for aquaculture, in ponds, lakes, and aquariums for monitoring growth, population size, and behavior traits.

• MeSH
• chemické látky znečišťující vodu metabolismus   MeSH
• kvalita vody MeSH
• monitorování životního prostředí metody   MeSH
• ryby metabolismus   MeSH
• severní raci metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• chemické látky znečišťující vodu MeSH

• MeSH
• diabetes mellitus 1. typu * MeSH
• krevní glukóza MeSH
• lidé MeSH
• novorozenec nedonošený MeSH
• novorozenec MeSH
• selfmonitoring glykemie * MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• Publikační typ
• časopisecké články MeSH
• komentáře MeSH
• Názvy látek
• krevní glukóza MeSH

PURPOSE: Herpes stromal keratitis is a serious condition and the most frequent cause of unilateral blindness. The real-time PCR is an accurate and fast diagnostic method for an analysis of infectious agents causing keratitis and keratouveitis. The aim of the study was to assess the relationship between clinical symptoms, treatment efficacy monitoring and viral quantity in corneal swabs determined by quantitative real-time PCR method. The real-time PCR method was used as well for the detection of other viral eye pathogens. METHODS: A total of 212 patients (136 men and 76 women) suspect of having herpes simplex virus (HSV) keratitis or keratouveitis were included in the study. The detection and quantitative analysis of the viral DNA were performed using the EliGene HSV1 RT kit, and the result was correlated with the clinical picture of the disease. The patients were routinely treated with acyclovir applied locally or, alternatively, in systemic administration. In a case of acyclovir treatment resistant keratitis, the patients were treated with local ganciclovir (Virgan gel ophth 0.15%). RESULTS: A total of 636 analyses of the viral DNA were performed; 85 patients were positive for HSV1 (198 detected). There were 16 acyclovir resistant cases of keratitis (14%). CONCLUSIONS: The real-time PCR appears as a fast and accurate method for an exact identification of the viral DNA in patients with herpes stromal keratitis. The introduction of the quantification is important for the treatment evaluation and for the specification of a so-called acyclovir resistant keratitis. A long-term systemic administration in maintenance doses may lead to the resistance and repeated, frequent relapses of the disease.

• MeSH
• acyklovir terapeutické užití   MeSH
• antivirové látky terapeutické užití   MeSH
• dítě MeSH
• DNA virů genetika   MeSH
• dospělí MeSH
• ganciklovir terapeutické užití   MeSH
• keratitida herpetická diagnóza   farmakoterapie   virologie   MeSH
• kvantitativní polymerázová řetězová reakce * MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidský herpesvirus 1 genetika   izolace a purifikace   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• uveitida diagnóza   farmakoterapie   virologie   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acyklovir MeSH
• antivirové látky MeSH
• DNA virů MeSH
• ganciklovir MeSH

A new setup for direct microspectroscopic monitoring of singlet oxygen ((1)O2) has been developed in our laboratory using a novel near-infrared sensitive InGaAs 2D-array detector. An imaging spectrograph has been inserted in front of the 2D-array detector, which allows us to acquire spectral images where one dimension is spatial and the other is spectral. The work presents a detailed examination of sensitivity and noise characteristics of the setup and its ability to detect (1)O2. The (1)O2 phosphorescence-based images and near-infrared luminescence spectral images recorded from single TMPyP-containing fibroblast cells reflecting spectral changes during irradiation are demonstrated. The introduction of spectral images addresses the issue of a potential spectral overlap of (1)O2 phosphorescence with near-infrared-extended luminescence of the photosensitizer and provides a powerful tool for distinguishing and separating them, which can be applied to any photosensitizer manifesting near-infrared luminescence.

• MeSH
• analýza jednotlivých buněk přístrojové vybavení   metody   MeSH
• buňky 3T3 MeSH
• fibroblasty metabolismus   MeSH
• fotochemické procesy MeSH
• fotosenzibilizující látky MeSH
• luminiscence MeSH
• mikrospektrofotometrie přístrojové vybavení   metody   MeSH
• myši MeSH
• počítačové systémy MeSH
• porfyriny MeSH
• singletový kyslík metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fotosenzibilizující látky MeSH
• porfyriny MeSH
• singletový kyslík MeSH
• tetra(4-N-methylpyridyl)porphine MeSH Prohlížeč

Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, is able to survive extreme conditions like very low pH (stomach), high temperature (pasteurization) or low temperature (refrigerated storage). Cheese, infant powder milk, cream and other milk and dairy products might thus be considered as possible sources of MAP for humans. The aim of this study was to investigate the survival of two MAP field isolates during fermentation of three different types of soured milk products (SMP; yogurt, acidophilus milk and kefir) under laboratory conditions. Pasteurized MAP-free milk was artificially contaminated with 10(6)MAPcells/mL and survival and absolute numbers of MAP were monitored during fermentation (4 or 16 h) and after six weeks of storage at 4°C by culture and quantitative real time PCR (qPCR). Viability of MAP was determined by culture using Herrold's egg yolk medium and Middlebrook 7H10 with antibiotics, supplemented with Mycobactin J and incubated at 37°C for up to 12 weeks. The absolute numbers of MAP were quantified by previously published qPCR assays targeting F57 and IS900 loci in MAP genome. We herein confirm that MAP can survive pH reduction, however, longer exposure to pH below 4 in SMP seems to be critical because it inhibits growth. Therefore, it is suggested that probiotic cultures that can decrease pH below 4 during fermentation could provide better inactivation of MAP in SMP.

• MeSH
• fermentace MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• mléčné výrobky mikrobiologie   MeSH
• mléko mikrobiologie   MeSH
• Mycobacterium avium subsp. paratuberculosis genetika   růst a vývoj   izolace a purifikace   MeSH
• paratuberkulóza mikrobiologie   MeSH
• pasterizace MeSH
• probiotika MeSH
• sýr MeSH
• vysoká teplota MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A rapid method for detection, discrimination and quantification of wheat and barley strains of wheat dwarf virus (WDV) was successfully developed. The sensitivity of quantification of the wheat and barley strains of WDV ranged from an average of 1.2 × 10(7)-1.2 × 10(2) and from an average of 1.4 × 10(7)-1.4 × 10(4) copies of viral genome, respectively. These standard serial dilutions were applied to plant and vector tissues for virus titer calculations. Both strains of WDV were clearly discriminated by specific probes and melting curve analysis. Both TaqMan(®) and SYBR(®) Green technologies provided accurate and reliable methods for monitoring, detection, discrimination, and quantification of WDV.

• MeSH
• Geminiviridae klasifikace   genetika   izolace a purifikace   MeSH
• ječmen (rod) virologie   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• nemoci rostlin virologie   MeSH
• pšenice virologie   MeSH
• senzitivita a specificita MeSH
• virová nálož metody   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH