The objective of this study was to evaluate hen performance and eggshell quality response to genotype, housing system, and feed calcium (Ca) level. For this purpose, an experiment was conducted on 360 laying hens of ISA Brown, Bovans Brown (commercial hybrids), and Moravia BSL (traditional Czech hybrid). Laying hens were kept in enriched cages and on littered floor and fed similar feed mixtures with different Ca content (3.00% vs. 3.50%). In terms of hen performance, ISA Brown had the highest egg production (84.2%) compared to Moravia BSL (74.3%) and Bovans Brown (71.4%). Regarding eggshell quality, Bovans Brown showed the highest values of all eggshell quality parameters. Increasing feed Ca level augmented egg production (p ≤ 0.001) but had no effect on other performance parameters. Except eggshell thickness, all eggshell quality parameters were affected by the three-way interaction of genotype, housing, and Ca. Bovans Brown, which had the strongest eggshells (5089 g/cm2) when housed on a littered floor system and fed 3.00% Ca, while Moravia BSL housed on a littered floor had the weakest eggshells (4236 g/cm2) at 3.50% Ca. The study pointed out the importance of the interactions between studied factors on performance and eggshell quality compared to an individual factor effect.

• Klíčová slova
• Ca, eggshell, enriched cages, genotype, hen performance, littered floor,
• Publikační typ
• časopisecké články MeSH

Genotyping of hepatitis C virus (HCV) is of relevance to scheduling the treatment of patients with chronic hepatitis C (VHC), making their prognosis and monitoring the treatment efficacy. A set of 62 sera testing HCV RNA positive in Cobas Amplicor HCV 2.0 test (CA) were genotyped using Versant HCV Genotype Assay (LiPA) Bayer, i.e. the reverse hybridization method, with the CA amplified product being directly used in the assay. Fifty-six out of 57 samples reactive in reverse hybridization (92%) were genotyped. One sample showed a profile differing from any genotype, five samples were not reactive and one sample was not tested within this study design. Two out of five non-reactive sera and one non-tested serum could be genotyped by nested PCR based reverse hybridization. It can be concluded that the CA product resulting from one-step HCV RNA amplification is suitable for use in genotyping by reverse hybridization. The CA product based genotyping procedure is easier to perform, less time-consuming and less costly. The nested PCR based procedure could be used for typing of sera with lower HCV concentrations nontypeable with the combination of CA and Versant HCV Genotype Assay. Forty-eight selected samples were typed not only by reverse hybridization but also by a serological kit Murex HCV Serotyping 1-6 Assay (Abbot Murex). Thirty-seven (77%) of these sera, including all of three sera negative in reverse hybridization, appeared typeable by this kit. Although less sensitive, serotyping may be of relevance to typing of sera with low HCV levels or not containing detectable viral NA which are nontypeable by reverse hybridization. Thirty-three sera appeared genotypeable by both of the methods tested with the results being in good agreement. In two cases only the serotyping method revealed one more type of virus (mixed genotype) compared to the reverse hybridization.

• MeSH
• chronická hepatitida C virologie   MeSH
• genotyp MeSH
• Hepacivirus klasifikace   genetika   MeSH
• hybridizace nukleových kyselin MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• RNA virová analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Názvy látek
• RNA virová MeSH

In order to find new informative predictors of myocardial infarction, we performed an analysis of genotype frequencies of polymorphic markers of SELE (rs2076059, 3832T > C), SELP (rs6131, S290 N), SELL (rs1131498, F206L), ICAM1 (rs5498, K469E), VCAM1 (rs3917010, c.928 + 420A > C), PECAM1 (rs668, V125L), VEGFA (rs35569394, -2549(18)I/D), CCL2 (rs1024611, -2518A > G), NOS3 (rs1799983, E298D), and DDAH1 (rs669173, c.303 + 30998A > G) genes in the group of Russian men with myocardial infarction (N = 315) and the control group of corresponding ethnicity, gender, and age (N = 286). Using Markov chain Monte-Carlo method (APSampler), we found genotype combinations associated with increased and decreased risk of myocardial infarction. The most significant associations were detected for PECAM1*V/V + DDAH1*C (OR = 4.17 CI 1.56-11.15 Pperm = 0.005) SELE*C + VEGFA*I + CCL2*G + VCAM1*A + NOS3*D (OR = 2.74 CI 1.66-4.52 Pperm = 2.09 × 10(-5)), and VEGFA*D/D + CCL2*A + DDAH1*C (OR = 0.44 CI 0.28-0.7 Pperm = 7.89 × 10(-5)) genotype combinations.

• Klíčová slova
• Genetic testing, Myocardial infarction, Risk prediction,
• MeSH
• amidohydrolasy genetika   MeSH
• antigeny CD31 genetika   MeSH
• dospělí MeSH
• frekvence genu MeSH
• genetická predispozice k nemoci MeSH
• genetické asociační studie MeSH
• genetické testování MeSH
• infarkt myokardu etnologie   genetika   MeSH
• jednonukleotidový polymorfismus MeSH
• lidé středního věku MeSH
• lidé MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Rusko MeSH
• Názvy látek
• amidohydrolasy MeSH
• antigeny CD31 MeSH
• dimethylargininase MeSH Prohlížeč

Out of three genotypes of Encephalitozoon cuniculi (I-III) available for experimental studies, E. cuniculi genotype I remains the less characterized. This study describes for the first time individual phases of microsporidiosis caused by E. cuniculi genotype I and efficacy of albendazole treatment in immunocompetent BALB/c and C57Bl/6 mice and immunodeficient SCID, CD4-/- and CD8-/- mice using molecular detection and quantification methods. We demonstrate asymptomatic infection despite an intense dissemination of microsporidia into most organs within the first weeks post infection, followed by a chronic infection characterized by significant microsporidia persistence in immunocompetent, CD4-/- and CD8-/- mice and a lethal outcome for SCID mice. Albendazole application led to loss E. cuniculi genotype I infection in immunocompetent mouse strains, decreased spore burden by half in CD4-/- and CD8-/- mice, and prolongation of survival of SCID mice. These results showed Encephalitozoon cuniculi genotype I infection extend and albendazole sensitivity was comparable to E. cuniculi genotype II, but the infection onset speed and mortality rate was similar to E. cuniculi genotype III. These imply that differences in the course of infection and the response to treatment depend not only on immunological status of the host, but also on the genotype causing the infection.

• Klíčová slova
• Encephalitozoon cuniculi genotype I, Experimental infection, Laboratory mice, RT PCR quantification,
• MeSH
• albendazol aplikace a dávkování   MeSH
• antigeny CD4 genetika   MeSH
• antigeny CD8 genetika   MeSH
• antiinfekční látky aplikace a dávkování   MeSH
• Encephalitozoon cuniculi klasifikace   genetika   MeSH
• encephalitozoonóza imunologie   parazitologie   MeSH
• genotyp MeSH
• imunokompetence MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši SCID MeSH
• myši MeSH
• polymerázová řetězová reakce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• albendazol MeSH
• antigeny CD4 MeSH
• antigeny CD8 MeSH
• antiinfekční látky MeSH

This study deals with detecting the associations of atopic dermatitis' (AD) phenotypes in children: alone or combined with seasonal allergic rhino-conjunctivitis (SARC) and/or perennial allergic rhinitis (PAR), and/or with bronchial asthma (BA) with single nucleotide polymorphisms (SNP) of filaggrin (FLG), thymic stromal lymphopoietin (TSLP) and orsomucoid-like-1 protein 3 (ORMDL3) genes. Male and female pediatric patients aged from 3 to 18 years old were recruited into the main (AD in different combinations with SARC, PAR, BA) and control groups (disorders of digestives system, neither clinical nor laboratory signs of atopy). Patients were genotyped for SNP of rs_7927894 FLG, rs_11466749 TSLP, rs_7216389 ORMDL3 variants. Statistically significant associations of the increased risk were detected of AD combined with SARC and/or PAR and AD combined with BA (possibly, SARC and/or PAR) with C/T rs_7927894 FLG and T/T rs_7216389 ORMDL3 genotypes. Genotype C/C rs_7927894 FLG significantly decreases the risk of AD combined with SARC and/or PAR by 2.56 fold. Several genotypes' associations had a trend to significance: C/C rs_7216389 ORMDL3 decreases and C/T rs_7216389 ORMDL3 increases the risk for developing AD alone phenotype; A/G rs_11466749 TSLP decreases the risk of AD combined with BA (possibly, SARC and/or PAR) phenotype development.

• Klíčová slova
• associations, atopic dermatitis, children, filaggrin, genotype, orsomucoid1-like protein 3, phenotype, thymic stromal lymphopoietin,
• MeSH
• atopická dermatitida genetika   MeSH
• celoroční alergická rýma genetika   MeSH
• cytokiny genetika   MeSH
• dítě MeSH
• fenotyp MeSH
• filagriny MeSH
• genetická predispozice k nemoci MeSH
• genotyp MeSH
• jednonukleotidový polymorfismus * MeSH
• kojenec MeSH
• konjunktivitida genetika   MeSH
• lidé MeSH
• membránové proteiny genetika   MeSH
• mladiství MeSH
• předškolní dítě MeSH
• proteiny S100 genetika   MeSH
• riziko MeSH
• rýma genetika   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cytokiny MeSH
• filagriny MeSH
• FLG protein, human MeSH Prohlížeč
• membránové proteiny MeSH
• ORMDL3 protein, human MeSH Prohlížeč
• proteiny S100 MeSH
• TSLP protein, human MeSH Prohlížeč

Of four genotypes of Encephalitozoon cuniculi, E. cuniculi genotype II is considered to represent a parasite that occurs in many host species in a latent asymptomatic form, whereas E. cuniculi genotype III seems to be more aggressive, and infections caused by this strain can lead to the death of even immunocompetent hosts. Although albendazole has been considered suitable for treatment of Encephalitozoon species, its failure in control of E. cuniculi genotype III infection has been reported. This study determined the effect of a 100× recommended daily dose of albendazole on an Encephalitozoon cuniculi genotype III course of infection in immunocompetent and immunodeficient mice and compared the results with those from experiments performed with a lower dose of albendazole and E. cuniculi genotype II. The administration of the regular dose of abendazole during the acute phase of infection reduced the number of affected organs in all strains of mice and absolute counts of spores in screened organs. However, the effect on genotype III was minor. Surprisingly, no substantial effect was recorded after the use of a 100× dose of albendazole, with larger reductions seen only in the number of affected organs and absolute counts of spores in all strains of mice, implying variations in albendazole resistance between these Encephalitozoon cuniculi genotypes. These results imply that differences in the course of infection and the response to treatment depend not only on the immunological status of the host but also on the genotype causing the infection. Understanding how microsporidia survive in hosts despite targeted antimicrosporidial treatment could significantly contribute to research related to human health.

• Klíčová slova
• Encephalitozoon cuniculi, albendazole, genotype III, microsporidiosis, tolerance, treatment,
• MeSH
• albendazol aplikace a dávkování   farmakologie   MeSH
• antifungální látky aplikace a dávkování   farmakologie   MeSH
• antigeny CD4 genetika   MeSH
• antigeny CD8 genetika   MeSH
• buněčné linie MeSH
• Cercopithecus aethiops MeSH
• Encephalitozoon cuniculi účinky léků   genetika   izolace a purifikace   MeSH
• encephalitozoonóza farmakoterapie   MeSH
• genotyp MeSH
• imunokompromitovaný pacient imunologie   MeSH
• mikrobiální testy citlivosti MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední BALB C MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši SCID MeSH
• myši MeSH
• počet mikrobiálních kolonií MeSH
• Vero buňky MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• albendazol MeSH
• antifungální látky MeSH
• antigeny CD4 MeSH
• antigeny CD8 MeSH

Understanding of the diversity of species of Cryptosporidium Tyzzer, 1910 in tortoises remains incomplete due to the limited number of studies on these hosts. The aim of the present study was to characterise the genetic diversity and biology of cryptosporidia in tortoises of the family Testudinidae Batsch. Faecal samples were individually collected immediately after defecation and were screened for presence of cryptosporidia by microscopy using aniline-carbol-methyl violet staining, and by PCR amplification and sequence analysis targeting the small subunit rRNA (SSU), Cryptosporidium oocyst wall protein (COWP) and actin genes. Out of 387 faecal samples from 16 tortoise species belonging to 11 genera, 10 and 46 were positive for cryptosporidia by microscopy and PCR, respectively. All samples positive by microscopy were also PCR positive. Sequence analysis of amplified genes revealed the presence of the Cryptosporidium tortoise genotype I (n = 22), C. ducismarci Traversa, 2010 (n = 23) and tortoise genotype III (n = 1). Phylogenetic analyses of SSU, COWP and actin gene sequences revealed that Cryptosporidium tortoise genotype I and C. ducismarci are genetically distinct from previously described species of Cryptosporidium. Oocysts of Cryptosporidium tortoise genotype I, measuring 5.8-6.9 µm × 5.3-6.5 µm, are morphologically distinguishable from C. ducismarci, measuring 4.4-5.4 µm × 4.3-5.3 µm. Oocysts of Cryptosporidium tortoise genotype I and C. ducismarci obtained from naturally infected Russian tortoises (Testudo horsfieldii Gray) were infectious for the same tortoise but not for Reeve's turtles (Mauremys reevesii [Gray]), common garter snake (Thamnophis sirtalis [Linnaeus]), zebra finches (Taeniopygia guttata [Vieillot]) and SCID mice (Mus musculus Linnaeus). The prepatent period was 11 and 6 days post infection (DPI) for Cryptosporidium tortoise genotype I and C. ducismarci, respectively; the patent period was longer than 200 days for both cryptosporidia. Naturally or experimentally infected tortoises showed no clinical signs of disease. Our morphological, genetic, and biological data support the establishment of Cryptosporidium tortoise genotype I as a new species, Cryptosporidium testudinis sp. n., and confirm the validity of C. ducismarci as a separate species of the genus Cryptosporidium.

• Klíčová slova
• molecular phylogeny, morphology, new species, taxonomy, transmission studies,
• MeSH
• Cryptosporidium klasifikace   genetika   MeSH
• feces parazitologie   MeSH
• fylogeneze * MeSH
• genotyp MeSH
• kryptosporidióza parazitologie   MeSH
• myši SCID MeSH
• myši MeSH
• ribozomální DNA genetika   MeSH
• želvy parazitologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ribozomální DNA MeSH

Microsporidia are obligate intracellurar unicellular parasite of wide range of vertebrates. Although ingestion or inhalation of microsporidian spores is the main route of infection, assumed vertical transmission was described in some mammals. The present study was focused on proof of vertical transmission in mice under experimental conditions. Mice were infected with E. cuniculi genotype II intraperitoneally after mating, or perorally followed by mating in acute or chronic phase of infection. Fetuses were delivered by Caesarean section or mice were kept up to the parturition. Some of cubs were immediately after birth transferred to uninfected surrogate mothers. Group of cubs was immunosuppressed. All cubs were examined using polymerase chain reaction for the presence of Encephalitozoon after birth or in their age of 3 or 6 weeks, respectively. All fetuses delivered by Caesarean section, which were intraperitoneally or perorally infected were negative as well as all neonatal mice and youngsters tested in age of 6 weeks. Only immunosuppressed cubs and cubs of immunodeficient mice in age of 21 days were positive for Encephalitozoon cuniculi genotype II. Present results provided the evidence that transplacental transmission of Encephalitozoon cuniculi in mice occurs, but the mechanism of these transport is still unknown.

• Klíčová slova
• BABL/c, Encephalitozoon cuniculi, Microsporidia, SCID, Transplacental transmission,
• MeSH
• Cercopithecus aethiops MeSH
• DNA fungální chemie   izolace a purifikace   MeSH
• Encephalitozoon cuniculi klasifikace   genetika   MeSH
• encephalitozoonóza imunologie   mikrobiologie   přenos   MeSH
• genotyp MeSH
• imunokompromitovaný pacient MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• spory hub MeSH
• těhotenství MeSH
• Vero buňky MeSH
• vertikální přenos infekce * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA fungální MeSH

Encephalitozoon cuniculi is probably the most common microsporidia which infects a wide range of vertebrates, including human. So far, four genotypes of this parasite have been identified based on the rRNA internal transcribed spacer variations. The course of infection caused by E. cuniculi III had very massive onset in immunocompetent host characterized by the presence of this parasite in all organs and tissues within one week after peroral infection. Encephalitozoonosis caused by E. cuniculi III had very progressive spreading into all organs within first week post inoculation in immunocompromised SCID mice and led to the death of the host. The experimental treatment with albendazole of immunocompetent BALB/c mice infected with E. cuniculi III have shown very weak effect. Our findings clearly showed that the different course of infection and response to treatment depends not only on the immunological status of the host, but also on the genotype of microsporidia. It could be very important especially for individuals under chemotherapy and transplant recipients of organs originating from infected donors.

• Klíčová slova
• Albendazole, BALB/C, Encephalitozoon cuniculi genotypes II and III, Microsporidiosis, SCID,
• MeSH
• albendazol farmakologie   terapeutické užití   MeSH
• antifungální látky farmakologie   terapeutické užití   MeSH
• Encephalitozoon cuniculi účinky léků   genetika   imunologie   fyziologie   MeSH
• encephalitozoonóza farmakoterapie   imunologie   parazitologie   MeSH
• feces parazitologie   MeSH
• genotyp MeSH
• imunokompetence * MeSH
• imunokompromitovaný pacient * MeSH
• myši inbrední BALB C MeSH
• myši SCID MeSH
• myši MeSH
• spory hub MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• albendazol MeSH
• antifungální látky MeSH

Chronic hepatitis C virus infection (HCV) has a negative impact on the long-term survival of recipients of kidney transplants. HCV should be treated in hemodialyzed patients before their enlistment for kidney transplantation in order to avoid the reactivation of virus after transplantation. Direct-acting antivirals represent the current standard of care in hemodialyzed patients with HCV genotypes 1 and 4; in patients with genotypes 2 or 3, the optimal regimen is yet to be established. Sofosbuvir (SOF) and daclatasvir (DCV) represent an antiviral pangenotypic regimen with favorable pharmacokinetics in hemodialyzed patients. We retrospectively evaluated safety and efficacy of the combination of SOF and DCV in the treatment of genotype 3a chronic HCV in six male patients (mean age of 39 years, range 25-53 years) with end-stage renal disease on maintenance hemodialysis; these patients were treated with a reduced dose of SOF (one half of a 400 mg tablet) and 60 mg of DCV once daily. The anticipated treatment duration was 12 weeks. Initial HCV RNA ranged from 120,000 to 11,000,000 IU/mL. Two of the six patients had compensated liver cirrhosis based on shear-wave elastography result. All of the patients completed a 12-week treatment. Viremia became negative on treatment and remained negative 12 weeks after the end of therapy in all the patients. All of them (6/6, 100%) achieved sustained virological response, including two with cirrhosis and two with HCV RNA >6,000,000 IU/mL. The treatment was well tolerated: none of the patients presented with a serious adverse event requiring hospital admission and none had anemia or any significant changes in blood count. One patient had a short period of diarrhea, which was resolved with antibiotic treatment. The combination of reduced-dose SOF and full-dose DCV, daily, was a safe and effective treatment in our group of hemodialyzed patients infected with HCV genotype 3.

• Klíčová slova
• HCV infection, daclatasvir, end-stage renal disease, genotype 3, maintenance hemodialysis, sofosbuvir,
• Publikační typ
• časopisecké články MeSH