Genotypizace viru hepatitidy C /HCV)má význam zejména pro stanovení schématu léčby pacientů s chronickou VHC a pro prognózu úspěšnosti léčby. U souboru 62 sér pozitivních na HCV RNA v testu Cobas Amplicor HCV 2.0 (CA) byl zjišťován genotyp HCV metodou reverzní hybridizace (Versant HCV Genotype Assay (LiPA) Bayer), modifikovanou tím způsobem, že jako výchozí amplifikovaný materiál pro reakci byl použit přímo produkt CA. Z 57 vzorků (92 %) reaktivních v reverzní hybridizaci byl u 56 určen genotyp. U jednoho vzorku neodpovídal profil žádnému určitému genotypu, pět vzorků bylo nereaktivních a jeden nebyl v tomto uspořádání testován. U dvou z těchto 5 nereaktivních a u jednoho netestovaného séra byl určen genotyp reverzní hybridizací vycházející z nested PCR. Lze shrnout, že produkt získaný po jednostupňové amplifikaci HCV RNA v testu CA je pro většinu vzorků vhodným výchozím materiálem pro genotypizaci reverzní hybridizací. Pro laboratoře používající k detekci HCV RNA v séru Cobas Amplicor HCV 2.0 by tento postup vedl ke zjednodušení genotypizace a k časové a materiálové úspoře. U sér s nižší koncentrací viru, která nejsou typizovatelná touto kombinací metod, lze vycházet z nested PCR. Čtyřicet osm vybraných vzorků bylo typizováno vedle reverzní hybridizace také sérologicky soupravou Murex HCV Serotyping 1–6 Assay (Abbot Murex). Typ byl určen u 37 ze 48 testovaných sér (77 %) včetně všech tří sér negativních v reverzní hybridizaci. Přestože je sérologická typizace méně citlivá, může být významná při typizaci některých sér s nízkou hladinou HCV RNA a sér neobsahujících již detekovatelnou virovou NK, která nejsou typizovatelná reverzní hybridizací. U 33 sér, jež byla genotypovatelná oběma metodami, byly tyto metody ve vzájemné shodě. Pouze ve dvou případech zjistila sérologická metoda přítomnost jednoho typu viru navíc (smíšený genotyp) oproti reverzní hybridizaci.

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
• hepatitida C virologie   MeSH
• lidé MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   využití   MeSH
• sérotypizace metody   využití   MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH

• MeSH
• finanční podpora výzkumu jako téma MeSH
• genetické techniky MeSH
• hyperlipoproteinemie typ II diagnóza   genetika   terapie   MeSH
• lidé MeSH
• Check Tag
• lidé MeSH

Úvod: Infekce virem hepatitidy C (HCV) je celosvětově jednou z hlavních příčin chronického onemocnění jater. Cíl: Retrospektivně zhodnotit výsledky léčby pacientů s infekcí HCV genotypu 2 na Klinice infekčních chorob LF MU a FN Brno během uplynulých 15 let. Pacienti a metodika: V průběhu 15 let bylo na naší klinice léčeno 15 pacientů (9 mužů, 6 žen, průměrný věk 54,73 ? 13,83 roku, medián 54 let) chronicky infikovaných HCV genotypu 2. Jedná se o největší soubor pacientů s touto diagnózou léčených v České republice. Zastoupení jednotlivých subtypů genotypu 2 HCV bylo následující: 1krát subtyp 2a, 2krát subtypy 2a/2c, 5krát subtyp 2b, ve zbylých 7 případech byl stanoven jen genotyp 2 bez možnosti určit subtyp. Výsledky: V letech 2002–2015 bylo 9 pacientů léčeno kombinací peginterferonu α-2a (180 ?g 1krát týdně podkožně) a ribavirinu (800 mg denně) po dobu 24 týdnů, setrvalé virologické odpovědi (SVR) dosáhlo 6 z nich, tedy 67 %. Ve 2 případech došlo po skončení léčby k relapsu, v jednom případě byl zaznamenán breakthrough fenomén. V letech 2015–2016 bylo 7 pacientů léčeno kombinací sofosbuviru (400 mg) a ribavirinu (1 000 nebo 1 200 mg denně podle hmotnosti) – 6krát se jednalo o první léčbu infekce HCV, 1krát o léčbu po breakthrough fenoménu vzniklého při předchozí léčbě. Všech 7 pacientů dosáhlo SVR (100 %). Závěr: Léčba infekce HCV genotypu 2 zatím nepředstavuje závažný terapeutický problém. V řadě rozvinutých zemí světa v posledních letech přibývá pacientů obtížně léčitelných, zejména cirhotiků. Léčebné možnosti se však rychle rozvíjejí.

Introduction: Hepatitis C virus (HCV) infection is one of the main causes of chronic liver disease worldwide. Aims: Retrospectively evaluate the results of therapy of patients with genotype 2 HCV treated during last 15 years at the Department of Infectious Diseases University Hospital Brno and Faculty of Medicine Masaryk University Brno, Czech Republic. Patients and methods: 15 patients (9 men, 6 women, mean age 54.73 ? 13.83 years, median 54 years) with chronic genotype 2 infection were treated at in our department during last 15 years. It is the biggest group of patients with this diagnosis treated in the Czech Republic. Proportion of genotype 2 subtypes was following: 1 times subtype 2a, 2 times subtype 2a/2c, 5 times subtype 2b, in the rest 7 cases only genotype 2 was determined without possibility to determinate subtype. Results: 9 patients were treated with the combination of peginterferon α-2a (180 ?g once weekly subcutaneously) and ribavirin (1 000 or 1 200 mg daily according to the weight) for 24 weeks in the years 2002–2015, sustained virological response (SVR) achieved 6 of them (67 %). Two relapses and one breakthrough were recorded. 7 patients were treated with combination of sofosbuvir (400 mg daily) and ribavirin (1 000 or 1 200 mg daily according to the weight) for 12 weeks in the years 2015–2016 – 6 of them were treatment na?ve, one after breakthrough in previous therapy. All of these 7 patients achieved SVR (100 %). Conclusions: Genotype 2 therapy still not represents serious therapeutic problem. The number of difficultly treatable patients is gradually increasing in many well-developed countries during last several years. However the therapeutic possibilities are quickly developing.

• Klíčová slova
• pegylovaný interferon,
• MeSH
• antivirové látky terapeutické užití   MeSH
• chronická hepatitida C * diagnóza   farmakoterapie   MeSH
• genotyp MeSH
• interferony aplikace a dávkování   terapeutické užití   MeSH
• kombinovaná farmakoterapie MeSH
• lidé MeSH
• retrospektivní studie MeSH
• ribavirin aplikace a dávkování   terapeutické užití   MeSH
• sofosbuvir aplikace a dávkování   terapeutické užití   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• dieta aterogenní MeSH
• dítě MeSH
• hyperlipoproteinemie typ II dietoterapie   farmakoterapie   genetika   MeSH
• kardiovaskulární nemoci prevence a kontrola   MeSH
• lidé MeSH
• mladiství MeSH
• mutační analýza DNA metody   využití   MeSH
• polymerázová řetězová reakce metody   využití   MeSH
• životní styl MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH

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.

• 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

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.

• 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

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.

• 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

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.

• 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

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.

• 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

Antibody (rituximab) dependent cellular cytotoxicity is a key mechanism in killing CD20+ lymphoma cells. FcγRIIIA-158 V/F gene polymorphism results in expression of 3 variants of the FcγRIIIA receptor (FcγRIIIA) on cytotoxic lymphocytes with different receptor affinity. We studied 102 patients with newly diagnosed FL to assess whether the FcγRIIIA genotype influences outcome in patients treated with risk-adapted immunochemotherapy. The median age was 52 years (31-84); 90% of the patients had advanced (III/IV) clinical stages. The Follicular Lymphoma International Prognostic Index (FLIPI) scores were as follows: low 18.9%, intermediate 33.7% and high 47.4%. The front-line treatment was stratified according to the commonly used risk factors (FLIPI, beta-2-microglobuline and serum-Tyrosine-Kinase levels, bulky disease) into 3 treatment groups: (1) patients with FLIPI 0-1 treated with (R)-CHOP (51%), (2) patients under 60 (65) years of age with intermediate-risk disease (FLIPI 2) indicated for an intensive protocol (ProMACE-CytaBOM or sequential chemotherapy) (21%), and (3) patients under 60 (65) years with high-risk disease (FLIPI ≥3) treated with intensive chemotherapy plus autologous stem cell transplantation (28%). Rituximab was added to front-line chemotherapy in 59% of the patients. Generally, complete remission (CR) or unconfirmed CR was achieved in 85% of the patients, 11% had partial remission and 4% stable disease. Molecular CR (CRm) was achieved in 67.4% of 86 evaluable patients. Overall survival (OS) at 5 years reached 84% (95% CI 0.74-0.93); event-free survival (EFS) at 5 years was 58% (95% CI 0.45-0.71). The frequencies of FcγRIIIA-158 gene polymorphisms V/V, V/F and F/F were 8%, 50% and 42%, respectively. The FLIPI score distribution was not different in F/F patients as compared to V/F+V/V carriers (chi-square, P=0.7). The treatment modalities (treatment arm or rituximab administration) had the same distribution in V/V+V/F vs F/F patients (chi-square, P=0.16 and P=0.62, respectively). The CRm rates were similar in both subgroups of V/V+V/F vs F/F patients (chi-square, P=0.92). Survival curves for OS and EFS were not significantly different when comparing the subgroups of V/V+V/F vs F/F patients (P=0.28 and P=0.57, respectively). We found no difference in the quality of treatment response or survival after front-line immunochemotherapy between FcγRIIIA subgroups. FcγRIIIA polymorphism have no influence on the outcome of patients treated with risk-adapted chemotherapy with or without rituximab.

• MeSH
• autologní transplantace MeSH
• dospělí MeSH
• folikulární lymfom genetika   mortalita   terapie   MeSH
• genotyp MeSH
• kombinovaná terapie MeSH
• lidé středního věku MeSH
• lidé MeSH
• multivariační analýza MeSH
• protokoly protinádorové kombinované chemoterapie terapeutické užití   MeSH
• receptory IgG genetika   MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• transplantace hematopoetických kmenových buněk MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH