P-glycoprotein (ABCB1), an ATP-binding-cassette efflux transporter, limits intestinal absorption of its substrates and is a common site of drug-drug interactions (DDIs). ABCB1 has been suggested to interact with many antivirals used to treat HIV and/or chronic hepatitis C virus (HCV) infections. Using bidirectional transport experiments in Caco-2 cells and a recently established ex vivo model of accumulation in precision-cut intestinal slices (PCIS) prepared from rat ileum or human jejunum, we evaluated the potential of anti-HIV and anti-HCV antivirals to inhibit intestinal ABCB1. Lopinavir, ritonavir, saquinavir, atazanavir, maraviroc, ledipasvir, and daclatasvir inhibited the efflux of a model ABCB1 substrate, rhodamine 123 (RHD123), in Caco-2 cells and rat-derived PCIS. Lopinavir, ritonavir, saquinavir, and atazanavir also significantly inhibited RHD123 efflux in human-derived PCIS, while possible interindividual variability was observed in the inhibition of intestinal ABCB1 by maraviroc, ledipasvir, and daclatasvir. Abacavir, zidovudine, tenofovir disoproxil fumarate, etravirine, and rilpivirine did not inhibit intestinal ABCB1. In conclusion, using recently established ex vivo methods for measuring drug accumulation in rat- and human-derived PCIS, we have demonstrated that some antivirals have a high potential for DDIs on intestinal ABCB1. Our data help clarify the molecular mechanisms responsible for reported increases in the bioavailability of ABCB1 substrates, including antivirals and drugs prescribed to treat comorbidity. These results could help guide the selection of combination pharmacotherapies and/or suitable dosing schemes for patients infected with HIV and/or HCV.
- Klíčová slova
- Caco-2 cells, P-glycoprotein, antiviral drugs, drug-drug interactions, intestinal absorption, precision-cut intestinal slices, rhodamine 123,
- MeSH
- antivirové látky farmakologie MeSH
- atazanavir sulfát farmakologie MeSH
- benzimidazoly farmakologie MeSH
- Caco-2 buňky účinky léků metabolismus MeSH
- fluoreny farmakologie MeSH
- hepatitida C komplikace farmakoterapie virologie MeSH
- HIV infekce komplikace farmakoterapie virologie MeSH
- imidazoly farmakologie MeSH
- karbamáty MeSH
- krysa rodu Rattus MeSH
- látky proti HIV farmakologie MeSH
- lékové interakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- Lopinavir farmakologie MeSH
- maravirok farmakologie MeSH
- P-glykoprotein antagonisté a inhibitory metabolismus MeSH
- potkani Wistar MeSH
- pyrrolidiny MeSH
- ritonavir farmakologie MeSH
- saquinavir farmakologie MeSH
- senioři MeSH
- střeva účinky léků MeSH
- valin analogy a deriváty MeSH
- zidovudin farmakologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antivirové látky MeSH
- atazanavir sulfát MeSH
- benzimidazoly MeSH
- daclatasvir MeSH Prohlížeč
- fluoreny MeSH
- imidazoly MeSH
- karbamáty MeSH
- látky proti HIV MeSH
- ledipasvir MeSH Prohlížeč
- Lopinavir MeSH
- maravirok MeSH
- P-glykoprotein MeSH
- pyrrolidiny MeSH
- ritonavir MeSH
- saquinavir MeSH
- valin MeSH
- zidovudin MeSH
- MeSH
- cílená molekulární terapie MeSH
- dospělí MeSH
- hepatitida C virologie MeSH
- imidazoly aplikace a dávkování MeSH
- lidé MeSH
- melanom farmakoterapie patologie MeSH
- metastázy nádorů MeSH
- nádory kůže farmakoterapie patologie MeSH
- oximy aplikace a dávkování MeSH
- poruchy spojené s užíváním amfetaminu rehabilitace MeSH
- protokoly antitumorózní kombinované chemoterapie aplikace a dávkování MeSH
- pyridony aplikace a dávkování MeSH
- pyrimidinony aplikace a dávkování MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- dopisy MeSH
- kazuistiky MeSH
- Názvy látek
- dabrafenib MeSH Prohlížeč
- imidazoly MeSH
- oximy MeSH
- pyridony MeSH
- pyrimidinony MeSH
- trametinib MeSH Prohlížeč
BACKGROUND: Treatment with direct acting antiviral agents (DAAs) has provided sustained virological response rates in >95% of patients with chronic hepatitis C virus (HCV) infection. However treatment is costly and market access, reimbursement and governmental restrictions differ among countries. We aimed to analyze these differences among European and Eurasian countries. METHODS: A survey including 20-item questionnaire was sent to experts in viral hepatitis. Countries were evaluated according to their income categories by the World Bank stratification. RESULTS: Experts from 26 countries responded to the survey. As of May 2016, HCV prevalence was reported as low (≤1%) in Croatia, Czech Republic, Denmark, France, Germany, Hungary, the Netherlands, Portugal, Slovenia, Spain, Sweden, UK; intermediate (1-4%) in Azerbaijan, Bosnia and Herzegovina, Italy, Kosovo, Greece, Kazakhstan, Romania, Russia, Serbia and high in Georgia (6.7%). All countries had national guidelines except Albania, Kosovo, Serbia, Tunisia, and UK. Transient elastography was available in all countries, but reimbursed in 61%. HCV-RNA was reimbursed in 81%. PegIFN/RBV was reimbursed in 54% of the countries. No DAAs were available in four countries: Kazakhstan, Kosovo, Serbia, and Tunisia. In others, at least one DAA combination with either PegIFN/RBV or another DAA was available. In Germany and the Netherlands all DAAs were reimbursed without restrictions: Sofosbuvir and sofosbuvir/ledipasvir were free of charge in Georgia. CONCLUSION: Prevalence of HCV is relatively higher in lower-middle and upper-middle income countries. DAAs are not available or reimbursed in many Eurasia and European countries. Effective screening and access to care are essential for reducing liver-related morbidity and mortality.
- Klíčová slova
- Availability of hepatitis C diagnostics, Therapeutics in European and Eurasia countries,
- MeSH
- antivirové látky terapeutické užití MeSH
- elastografie MeSH
- Hepacivirus * MeSH
- hepatitida C diagnóza farmakoterapie epidemiologie virologie MeSH
- lidé MeSH
- prevalence MeSH
- úhrada zdravotního pojištění MeSH
- virová nálož MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Asie epidemiologie MeSH
- Evropa epidemiologie MeSH
- Názvy látek
- antivirové látky MeSH
UNLABELLED: Alternations in the glycosylation of proteins have been described in connection with several cancers, including hepatocellular carcinoma (HCC) and colorectal cancer. Analytical tools, which use combination of liquid chromatography and mass spectrometry, allow precise and sensitive description of these changes. In this study, we use MRM and FT-ICR operating in full-MS scan, to determine ratios of intensities of specific glycopeptides in HCC, colorectal cancer, and liver metastasis of colorectal cancer. Haptoglobin, hemopexin and complement factor H were detected after albumin depletion and the N-linked glycopeptides with fucosylated glycans were compared with their non-fucosylated forms. In addition, sialylated forms of an O-linked glycopeptide of hemopexin were quantified in the same samples. We observe significant increase in fucosylation of all three proteins and increase in bi-sialylated O-glycopeptide of hemopexin in HCC of hepatitis C viral (HCV) etiology by both LC-MS methods. The results of the MRM and full-MS scan FT-ICR analyses provide comparable quantitative readouts in spite of chromatographic, mass spectrometric and data analysis differences. Our results suggest that both workflows allow adequate relative quantification of glycopeptides and suggest that HCC of HCV etiology differs in glycosylation from colorectal cancer and liver metastasis of colorectal cancer. SIGNIFICANCE: The article compares N- and O-glycosylation of several serum proteins in different diseases by a fast and easy sample preparation procedure in combination with high resolution Fourier transform ion cyclotron resonance mass spectrometry. The results show successful glycopeptides relative quantification in a complex peptide mixture by the high resolution instrument and the detection of glycan differences between the different types of cancer diseases. The presented method is comparable to conventional targeted MRM approach but allows additional curation of the data.
- Klíčová slova
- FT-ICR, Glycomics, Haptoglobin, Hemopexin, Hepatocellular carcinoma, MRM, Mass spectrometry, N-glycopeptides, O-glycopeptides, Quantification,
- MeSH
- cyklotrony MeSH
- diferenciální diagnóza MeSH
- Fourierova analýza MeSH
- glykopeptidy analýza MeSH
- glykosylace MeSH
- hepatitida C komplikace virologie MeSH
- hepatocelulární karcinom diagnóza etiologie sekundární MeSH
- hmotnostní spektrometrie přístrojové vybavení metody MeSH
- kolorektální nádory diagnóza etiologie patologie MeSH
- lidé MeSH
- nádory jater diagnóza etiologie sekundární MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- glykopeptidy MeSH
BACKGROUND: Sequence variability in the hepatitis C virus (HCV) genome has led to the development and classification of six genotypes and a number of subtypes. The HCV 5' untranslated region mainly comprises an internal ribosomal entry site (IRES) responsible for cap-independent synthesis of the viral polyprotein and is conserved among all HCV genotypes. DESCRIPTION: Considering the possible high impact of variations in HCV IRES on viral protein production and thus virus replication, we decided to collect the available data on known nucleotide variants in the HCV IRES and their impact on IRES function in translation initiation. The HCV IRES variation database (HCVIVdb) is a collection of naturally occurring and engineered mutation entries for the HCV IRES. Each entry contains contextual information pertaining to the entry such as the HCV genotypic background and links to the original publication. Where available, quantitative data on the IRES efficiency in translation have been collated along with details on the reporter system used to generate the data. Data are displayed both in a tabular and graphical formats and allow direct comparison of results from different experiments. Together the data provide a central resource for researchers in the IRES and hepatitis C-oriented fields. CONCLUSION: The collation of over 1900 mutations enables systematic analysis of the HCV IRES. The database is mainly dedicated to detailed comparative and functional analysis of all the HCV IRES domains, which can further lead to the development of site-specific drug designs and provide a guide for future experiments. HCVIVdb is available at http://www.hcvivdb.org .
- Klíčová slova
- Database, HCV, Hepatitis C, IRES, Internal ribosome entry site, Translation efficiency,
- MeSH
- 5' nepřekládaná oblast MeSH
- databáze genetické * MeSH
- genotyp MeSH
- Hepacivirus genetika metabolismus MeSH
- hepatitida C virologie MeSH
- IRES genetika MeSH
- lidé MeSH
- mutace MeSH
- proteosyntéza MeSH
- RNA virová genetika MeSH
- sběr dat MeSH
- sekvence nukleotidů MeSH
- virové proteiny biosyntéza genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 5' nepřekládaná oblast MeSH
- IRES MeSH
- RNA virová MeSH
- virové proteiny MeSH
OBJECTIVES: The aim was to introduce a diagnostic method for detecting variants of hepatitis C virus (HCV) with protease NS3 resistance primarily to simeprevir (Q80K mutation in HCV genotype 1a) and its subsequent use in routine practice. MATERIAL AND METHODS: The detection of HCV resistance-associated variants in the NS3 protease gene by sequence analysis was introduced in the molecular biology laboratory of University Hospital Hradec Kralove in 2015. The primers were designed by sequence analysis software Custom Primers - OligoPerfect™ Designer. The method was optimized for HCV genotype 1a. The search for variants was performed using two programs. RESULTS: A total of 16 patients with genotype 1a chronic hepatitis C have been examined since 2015. In five of them, the Q80K variant was detected. CONCLUSION: The development of resistance to antiviral therapy for chronic hepatitis C gained importance after the introduction of direct-acting antivirals. Given the relatively high prevalence of the Q80K mutation in HCV genotype 1a, it is crucial to confirm its presence or absence before the therapy is initiated. The reported method enables clear and early detection of the Q80K mutation.
- MeSH
- Hepacivirus genetika MeSH
- hepatitida C diagnóza virologie MeSH
- lidé MeSH
- mutace genetika MeSH
- virologie metody MeSH
- virová léková rezistence genetika MeSH
- virové nestrukturální proteiny genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- NS3 protein, hepatitis C virus MeSH Prohlížeč
- virové nestrukturální proteiny MeSH
The pathogenesis of hepatitis C virus (HCV) infection is regulated by the host immunity and several metabolic factors affecting liver metabolism, including oxidative stress, insulin resistance, and hepatic steatosis. Both innate and adaptive immunity play an important role in HCV infection. Cytotoxic lymphocytes have a crucial role in viral eradication or viral persistence. Major cause of viral persistence during HCV infection could be the development of a weak antiviral immune response to the viral antigens, with corresponding inability to eradicate infected cells.
- Klíčová slova
- hepatitis C virus - pathogenesis - immunity - Th1 immune response - Th2 immune response - regulatory T cells - Tregs/Th17 ratio - IFN-γ - TNF-α - IL-2 - IL-10 - TGF-β.,
- MeSH
- cytotoxické T-lymfocyty imunologie MeSH
- Hepacivirus genetika imunologie patogenita MeSH
- hepatitida C imunologie virologie MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
BACKGROUND: Liver stiffness determined by transient elastography is correlated with hepatic fibrosis stage and has high accuracy for detecting severe fibrosis and cirrhosis in chronic hepatitis C patients. We evaluated the clinical value of baseline FibroScan values for the prediction of safety and efficacy of telaprevir-based therapy in patients with advanced fibrosis and cirrhosis in the telaprevir Early Access Program HEP3002. METHODS: 1,772 patients with HCV-1 and bridging fibrosis or cirrhosis were treated with telaprevir plus pegylated interferon-α and ribavirin (PR) for 12 weeks followed by PR alone, the total treatment duration depending on virological response and previous response type. Liver fibrosis stage was determined either by liver biopsy or by non-invasive markers. 1,282 patients (72%) had disease stage assessed by FibroScan; among those 46% were classified as Metavir F3 at baseline and 54% as F4. RESULTS: Overall, 1,139 patients (64%) achieved a sustained virological response (SVR) by intention-to-treat analysis. Baseline FibroScan values were tested for association with SVR and the occurrence of adverse events. By univariate analysis, higher baseline FibroScan values were predictive of lower sustained virological response rates and treatment-related anemia. By multivariate analysis, FibroScan was no longer statistically significant as an independent predictor, but higher FibroScan values were correlated with the occurrence of infections and serious adverse events. CONCLUSIONS: FibroScan has a limited utility as a predictor of safety and efficacy in patients treated with telaprevir-based triple therapy. Nevertheless it can be used in association with other clinical and biological parameters to help determine patients who will benefit from the triple regiments. TRIAL REGISTRATION: ClinicalTrials.gov NCT01508286.
- MeSH
- analýza podle původního léčebného záměru MeSH
- anemie etiologie MeSH
- biomechanika účinky léků MeSH
- dospělí MeSH
- dostupnost zdravotnických služeb MeSH
- Hepacivirus účinky léků fyziologie MeSH
- hepatitida C komplikace farmakoterapie patofyziologie virologie MeSH
- jaterní cirhóza komplikace farmakoterapie patofyziologie MeSH
- játra účinky léků patologie patofyziologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- multivariační analýza MeSH
- oligopeptidy škodlivé účinky farmakologie terapeutické užití MeSH
- rizikové faktory MeSH
- senioři MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- práce podpořená grantem MeSH
- Názvy látek
- oligopeptidy MeSH
- telaprevir MeSH Prohlížeč
The Roche Cobas TaqScreen MPX Test v1 - multiplex reverse transcription-real time (MPX RT-Real Time) PCR, performed on Cobas s201 for HCV RNA, HBV DNA, HIV-1 RNA /group M and O/, and HIV-2 RNA was introduced as a supplement to the currently used imunoanalysis method for blood donor´s testing (Abbott CMIA - chemiluminescent microparticle imunoassay, performed on Architect i2000 for anti-HCV, hepatitis B surface antigen (HBsAg)), anti-HIV-1 /group M and O/, anti-HIV-2 and p24 HIV). The results of study could provide valuable arguments to support the discussion about the NAT implementation into the standards of blood donor´s testing in the Czech Republic. Two groups of samples were tested. In the first one, 5074 samples from consecutive blood donors, and in the second one, 5 repository preseroconverted samples from repeat blood donors, who were subsequently confirmed positive for Viral Hepatitis and/or HIV/AIDS by the National Reference Laboratory (NRL), were tested. One sample was found reactive by chemiluminescent microparticle immuno assay (CMIA) and nucleic acid test (NAT) (confirmed HBV-positive in NRL), 31 samples were CMIA-only reactive (15 anti-HCV, 4 HBsAg, 12 anti-HIV/p24, all confirmed negative in NRL) and one pool (6 samples) was found reactive (further individual NAT was negative for all samples) in the first group of samples. One sample was NAT-only reactive (confirmed HCV-positive in NRL) in the second group of samples. Our study confirmed that screening of infectious markers using NAT can reduce the risk of transmitting the monitored infections by blood transfusion in the Czech Republic, even as a country with currently good epidemiological situation.
- MeSH
- Hepacivirus genetika izolace a purifikace MeSH
- hepatitida B diagnóza epidemiologie virologie MeSH
- hepatitida C diagnóza epidemiologie virologie MeSH
- HIV infekce diagnóza epidemiologie virologie MeSH
- HIV-1 genetika izolace a purifikace MeSH
- HIV-2 genetika izolace a purifikace MeSH
- lidé MeSH
- virus hepatitidy B genetika izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
BACKGROUND AND AIMS: Hepatitis C virus (HCV) infection is associated with systemic oxidative stress. Since the heme catabolic pathway plays an important role in antioxidant protection, we attempted to assess the gene expression of key enzymes of heme catabolism, heme oxygenase 1 (HMOX1), heme oxygenase 2 (HMOX2), and biliverdin reductase A (BLVRA) in the liver and peripheral blood leukocytes (PBL) of patients chronically infected with HCV. METHODS: Gene expressions (HMOX1, HMOX2, BLVRA) and HCV RNA were analyzed in PBL of HCV treatment naïve patients (n = 58) and controls (n = 55), with a subset of HCV patients having data on hepatic gene expression (n = 35). Based upon the therapeutic outcome, HCV patients were classified as either responders (n = 38) or treatment-failure patients (n = 20). Blood samples in HCV patients were collected at day 0, and week 12, 24, 36, and 48 after the initiation of standard antiviral therapy. RESULTS: Compared to the controls, substantially increased BLVRA expression was detected in PBL (p<0.001) of therapeutically naïve HCV patients. mRNA levels of BLVRA in PBL closely correlated with those in liver tissue (r2 = 0.347,p = 0.03). A marked difference in BLVRA expression in PBL between the sustained responders and patients with treatment failure was detected at week 0 and during the follow-up (p<0.001). Multivariate analysis revealed that BLVRA basal expression in PBL was an independent predictor for sustained virological response (OR 15; 95% CI 1.05-214.2; P = 0.046). HMOX1/2 expression did not have any effect on the treatment outcome. CONCLUSION: Our results suggest that patients with chronic HCV infection significantly upregulate BLVRA expression in PBL. The lack of BLVRA overexpression is associated with non-responsiveness to standard antiviral therapy; whereas, HMOX1/2 does not seem to have any predictive potential.
- MeSH
- antivirové látky terapeutické užití MeSH
- dospělí MeSH
- exprese genu * MeSH
- hem metabolismus MeSH
- hemová oxygenasa (decyklizující) genetika metabolismus MeSH
- hemoxygenasa-1 genetika metabolismus MeSH
- Hepacivirus * MeSH
- hepatitida C farmakoterapie genetika virologie MeSH
- játra metabolismus virologie MeSH
- leukocyty metabolismus virologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- metabolické sítě a dráhy genetika MeSH
- oxidoreduktasy působící na CH-CH vazby genetika MeSH
- ROC křivka MeSH
- výsledek terapie 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
- biliverdin reductase MeSH Prohlížeč
- hem MeSH
- heme oxygenase-2 MeSH Prohlížeč
- hemová oxygenasa (decyklizující) MeSH
- hemoxygenasa-1 MeSH
- oxidoreduktasy působící na CH-CH vazby MeSH
