A thermoresponsive Pluronic/alginate semisynthetic hydrogel is used to bioprint 3D hepatic constructs, with the aim to investigate liver-specific metabolic activity of the 3D constructs compared to traditional 2D adherent cultures. The bioprinting method relies on a bioinert hydrogel and is characterized by high-shape fidelity, mild depositing conditions and easily controllable gelation mechanism. Furthermore, the dissolution of the sacrificial Pluronic templating agent significantly ameliorates the diffusive properties of the printed hydrogel. The present findings demonstrate high viability and liver-specific metabolic activity, as assessed by synthesis of urea, albumin, and expression levels of the detoxifying CYP1A2 enzyme of cells embedded in the 3D hydrogel system. A markedly increased sensitivity to a well-known hepatotoxic drug (acetaminophen) is observed for cells in 3D constructs compared to 2D cultures. Therefore, the 3D model developed herein may represent an in vitro alternative to animal models for investigating drug-induced hepatotoxicity.

• Keywords
• 3D liver models, Pluronic/alginate thermogels, bioprinting, drug hepatotoxicity, hepatic constructs,
• MeSH
• Printing, Three-Dimensional MeSH
• Bioprinting * MeSH
• Hydrogels MeSH
• Chemical and Drug Induced Liver Injury * MeSH
• Tissue Engineering MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Hydrogels MeSH

Nephronophthisis-related ciliopathies (NPHP-RC) are recessive diseases characterized by renal dysplasia or degeneration. We here identify mutations of DCDC2 as causing a renal-hepatic ciliopathy. DCDC2 localizes to the ciliary axoneme and to mitotic spindle fibers in a cell-cycle-dependent manner. Knockdown of Dcdc2 in IMCD3 cells disrupts ciliogenesis, which is rescued by wild-type (WT) human DCDC2, but not by constructs that reflect human mutations. We show that DCDC2 interacts with DVL and DCDC2 overexpression inhibits β-catenin-dependent Wnt signaling in an effect additive to Wnt inhibitors. Mutations detected in human NPHP-RC lack these effects. A Wnt inhibitor likewise restores ciliogenesis in 3D IMCD3 cultures, emphasizing the importance of Wnt signaling for renal tubulogenesis. Knockdown of dcdc2 in zebrafish recapitulates NPHP-RC phenotypes, including renal cysts and hydrocephalus, which is rescued by a Wnt inhibitor and by WT, but not by mutant, DCDC2. We thus demonstrate a central role of Wnt signaling in the pathogenesis of NPHP-RC, suggesting an avenue for potential treatment of NPHP-RC.

• MeSH
• Adaptor Proteins, Signal Transducing genetics   metabolism   MeSH
• beta Catenin antagonists & inhibitors   metabolism   MeSH
• NIH 3T3 Cells MeSH
• Cilia genetics   pathology   MeSH
• Kidney Diseases, Cystic genetics   MeSH
• Zebrafish genetics   MeSH
• Exons MeSH
• Phenotype MeSH
• Phosphoproteins genetics   metabolism   MeSH
• HEK293 Cells MeSH
• Kidney pathology   MeSH
• Humans MeSH
• Mutation MeSH
• Mice MeSH
• Dishevelled Proteins MeSH
• Microtubule-Associated Proteins genetics   metabolism   MeSH
• Wnt Signaling Pathway genetics   MeSH
• Microscopy, Electron, Transmission MeSH
• Computational Biology MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Names of Substances
• Adaptor Proteins, Signal Transducing MeSH
• beta Catenin MeSH
• DCDC2 protein, human MeSH Browser
• DCDC2 protein, mouse MeSH Browser
• Phosphoproteins MeSH
• Dishevelled Proteins MeSH
• Microtubule-Associated Proteins MeSH

Expression system based on vaccinia virus (VV) is used both for recombinant protein production in vitro and as an alive vaccine. The article summarizes various strategies for recombinant VV construction, and describes preparation of recombinants expressing various forms of S and C genes of hepatitis B virus (HBV). It is shown, that the CV-1 cells infected with these recombinants synthesized the MS (middle) and the LS (large) polypeptides of the surface antigen (HBsAg) and the nucleocapsid antigen (HBcAg) polypeptide and the polypeptide HBeAg. Posttranslation modification of all expressed proteins was the same as at HBV infection.

• MeSH
• Hepatitis B genetics   MeSH
• Recombination, Genetic MeSH
• Recombinant Proteins biosynthesis   MeSH
• Vaccinia virus * MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH
• Names of Substances
• Recombinant Proteins MeSH

Chronic hepatitis B (CHB) is caused by the Hepatitis B virus (HBV) and affects millions of people worldwide. Developing an effective CHB therapy requires using in vivo screening methods, such as mouse models reflecting CHB based on hydrodynamic delivery of plasmid vectors containing a replication-competent HBV genome. However, long-term expression of HBV proteins is accompanied by production of progeny virions, thereby requiring a Biosafety Level (BSL) 3 animal facility. In the present study, we introduced a point mutation in the START codon of the HBV polymerase to develop a mouse model reflecting chronic hepatitis B infection without formation of viral progeny. We induced the mouse model by hydrodynamic injection of adeno-associated virus plasmid vector (pAAV) and minicircle plasmid (pMC) constructs into C57Bl/6 and C3H/HeN mouse strains, monitoring HBV antigens and antibodies in blood by enzyme-linked immunosorbent assay and analyzing liver expression of HBV core antigen by immunohistology. Persisting expression of viral antigens over 140 days (study endpoint) was observed only in the C3H/HeN mouse strain when using pAAV/1.2HBV-A and pMC/1.0HBV-D with pre-C and pre-S recombination sites. In addition, pAAV/1.2HBV-A in C3H/HeN sustained HBV core antigen positivity up to the study endpoint in C3H/HeN mice. Moreover, introducing the point mutation in the START codon of polymerase effectively prevented the formation of viral progeny. Our study establishes an accessible and affordable experimental paradigm for developing a robust mouse model reflecting CHB suitable for preclinical testing of anti-HBV therapeutics in a BSL2 animal facility.

• MeSH
• Hepatitis B, Chronic * genetics   MeSH
• Codon, Initiator MeSH
• Disease Models, Animal MeSH
• Mutation MeSH
• Mice, Inbred C3H MeSH
• Mice MeSH
• Hepatitis B virus genetics   MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Codon, Initiator MeSH
• P protein, Hepatitis B virus MeSH Browser

BACKGROUND: Following the introduction of direct-acting antiviral therapy in 2013, WHO launched the first Global Health Sector Strategy on Viral Hepatitis. We describe a hepatitis C virus (HCV) cascade of care in people with HIV (PWH) across Europe in terms of reaching the WHO elimination targets of diagnosing 90% and treating 80% of HCV-infected individuals. METHODS: HIV/HCV-coinfected participants in the EuroSIDA cohort under prospective follow-up at October 1, 2019, were described using a nine-stage cascade of care. Care cascades were constructed across Europe, on a regional (n = 5) and country (n = 21) level. RESULTS: Of 4773 anti-HCV positive PWH, 4446 [93.1%, 95% confidence interval (CI) 92.4-93.9)] were ever tested for HCV RNA, and 19.0% (95% CI 16.4-21.6) were currently HCV RNA positive, with the highest prevalence in Eastern and Central-Eastern Europe (33.7 and 29.6%, respectively). In Eastern Europe, 78.1% of the estimated number of chronic infections have been diagnosed, whereas this proportion was above 95% in the other four regions. Overall, 3116 persons have ever started treatment (72.5% of the ever chronically infected, 95% CI 70.9-74.0) and 2404 individuals (55.9% of the ever chronically infected, 95% CI 53.9-57.9) were cured. Cure proportion ranged from 11.2% in Belarus to 87.2% in Austria. CONCLUSION: In all regions except Eastern Europe, more than 90% of the study participants have been tested for HCV-RNA. In Southern and Central-Western regions, more than 80% ever chronically HCV-infected PWH received treatment. The proportion with cured HCV infection did not exceed 80% in any region, with significant heterogeneity between countries. SUMMARY: In a pan-European cohort of PWH, all regions except Eastern Europe achieved the WHO target of diagnosing 90% of chronic HCV infections, while the target of treating 80% of eligible persons was achieved in none of the five regions.

• MeSH
• Antiviral Agents therapeutic use   MeSH
• Hepatitis C, Chronic * complications   drug therapy   epidemiology   MeSH
• Hepacivirus genetics   MeSH
• Hepatitis C * complications   drug therapy   epidemiology   MeSH
• HIV Infections * complications   drug therapy   epidemiology   MeSH
• Coinfection * drug therapy   MeSH
• Humans MeSH
• Prospective Studies MeSH
• RNA therapeutic use   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Europe epidemiology   MeSH
• Names of Substances
• Antiviral Agents MeSH
• RNA MeSH

One of the therapeutic approaches in acute liver failure is the use of an artificial system replacing hepatic function--bioartificial liver. Its application is the most perspective in fulminant liver failure during preparation for transplantation of the liver (so-called bridge to transplantation) or in case of a non-functioning hepatic graft, and to reduce the mortality and morbidity of patients with acute liver failure where transplantation is not indicated or where a suitable graft was not found. The authors discuss briefly the construction of these systems, analyze different indications of treatment and its results, obscure questions and perspectives of further development.

• MeSH
• Liver Failure, Acute therapy   MeSH
• Liver, Artificial * MeSH
• Humans MeSH
• Check Tag
• Humans MeSH
• Publication type
• English Abstract MeSH
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Rapid and sensitive detection is a key step in the effective and early response to the global hazard of various viral diseases. In this study, an integrated isolation of hepatitis B virus (HBV)-specific DNA fragment by magnetic nanoparticles (MNPs) and its immediate analysis by microchip CGE was performed. Microfluidic CE chip was used to accommodate the complete process of viral DNA isolation by MNPs including hybridization and thermal denaturation followed by CE separation. Beforehand, calibration curves of HBV fragments were constructed. For isolation by MNPs, specific streptavidin-biotin interaction was used to bind complementary HBV fragment to magnetic particles. After analysis of isolated HBV by regular MNPs method, innovative approach was performed. The commercial CE chip (Bio-rad) was successfully used to execute HBV fragment isolation. Detection using LIF with detection limit of 1 ng/mL was accomplished.

• MeSH
• DNA, Viral isolation & purification   MeSH
• Electrophoresis, Microchip instrumentation   MeSH
• Hepatitis B diagnosis   MeSH
• Humans MeSH
• Limit of Detection MeSH
• Magnetite Nanoparticles chemistry   MeSH
• Oligonucleotides isolation & purification   MeSH
• Hepatitis B virus isolation & purification   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• DNA, Viral MeSH
• Magnetite Nanoparticles MeSH
• Oligonucleotides MeSH

Symptomatic hepatic porphyria was followed up in workers exposed to the risk of free SiO2. This disease was found in 12 out of 998 workers engaged in the construction of the underground and exposed to SiO2 (group I), and in 12 at-risk persons out of 440 patients from different workplaces who were hospitalized for SiO2 exposure (group II). The average age of the workers of group I was 40.6 +/- 7.7, that of the group II 54.9 +/- 13.3. In both these high-risk groups, porphyria was found to be significantly more frequent than in the control group not exposed to free SiO2. The authors discuss the long-lasting effects of free SiO2 on the occurrence of symptomatic hepatic porphyria.

• MeSH
• Adult MeSH
• Chemical and Drug Induced Liver Injury * MeSH
• Middle Aged MeSH
• Humans MeSH
• Occupational Diseases chemically induced   MeSH
• Silicon Dioxide adverse effects   MeSH
• Porphyrias chemically induced   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Silicon Dioxide MeSH

Five different hybridoma clones secreting anti-HBeAg antibody were constructed by fusing cells of mouse myeloma line SP2/0 with splenocytes from BALB/c mice immunized with recombinant HBeAg. The monoclonal antibodies obtained were characterized immunologically and one was used to develop ELISA for detection of HBeAg and anti-HBeAg antibody. These monoclonal assays enabled the detection of 3 U HBeAg/ml and 1 U anti-HBeAg/ml with reference to standards of the Paul Ehrlich Institute, Frankfurt, F.R.G. Both assays compared well with a commercially available kit (Abbott Laboratory) and were used for detection of HBeAg and anti-HBeAg antibody in clinical serum samples.

• MeSH
• Cell Line MeSH
• Hepatitis B e Antigens immunology   MeSH
• Hepatitis B Surface Antigens immunology   MeSH
• Hepatitis B Antibodies analysis   MeSH
• Hepatitis B diagnosis   MeSH
• Hybridomas immunology   MeSH
• Humans MeSH
• Antibodies, Monoclonal immunology   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Recombinant Proteins immunology   MeSH
• Hepatitis B virus immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Hepatitis B e Antigens MeSH
• Hepatitis B Surface Antigens MeSH
• Hepatitis B Antibodies MeSH
• Antibodies, Monoclonal MeSH
• Recombinant Proteins MeSH

Vaccinia virus (VV) recombinants expressing hepatitis B virus (HBV) surface (HBsAg) or core (HBcAg) antigens (Kunke et al., Virology 195, 132 - 139 (1993)] have been shown to raise specific antibodies in mice, nevertheless the levels of antibodies reactive with the preS2 and S antigens were low. In an attempt to enhance the immunogenicity of HBsAg-preS2, a fused C-preS2 gene was constructed. The fusion protein was expressed in E. coli and displayed both HBcAg and preS2 antigen as demonstrated by enzyme-linked immunosorbent assay (ELISA). The same gene was then expressed using recombinant VV and chimerical particles whose size and density were similar to those of native HBV core particles produced in CV-1 cells infected with recombinant VV. Unlike HBcAg, preS2 antigen could not be detected on these particles by ELISA but was revealed by immunoblot analysis only. The immunogenicity of the recombinant VV was evaluated in mice. Antibodies to HBcAg and VV antigen but not to preS2 antigen were found in sera of animals inoculated with 10(7) PFU of the recombinant VV. Presumably, HBcAg-preS2 particles produced in E. coli and in eukaryotic cells have a different conformation, and the presence of preS2 antigen on the surface of chimerical particle might be necessary for a pronounced antibody response.

• MeSH
• Cell Line MeSH
• Chlorocebus aethiops MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Escherichia coli MeSH
• Genetic Vectors MeSH
• Hepatitis B Core Antigens biosynthesis   genetics   immunology   MeSH
• Hepatitis B Surface Antigens biosynthesis   genetics   immunology   MeSH
• Hepatitis B Antigens biosynthesis   genetics   immunology   MeSH
• Immunoblotting MeSH
• Immunogenetics MeSH
• Rats MeSH
• Humans MeSH
• Mice, Inbred ICR MeSH
• Mice MeSH
• Protein Precursors biosynthesis   genetics   immunology   MeSH
• Recombinant Fusion Proteins biosynthesis   genetics   immunology   MeSH
• Vaccines, Synthetic immunology   MeSH
• Hepatitis B Vaccines immunology   MeSH
• Hepatitis B virus immunology   MeSH
• Vaccinia virus MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Hepatitis B Core Antigens MeSH
• Hepatitis B Surface Antigens MeSH
• Hepatitis B Antigens MeSH
• presurface protein 2, hepatitis B surface antigen MeSH Browser
• Protein Precursors MeSH
• Recombinant Fusion Proteins MeSH
• Vaccines, Synthetic MeSH
• Hepatitis B Vaccines MeSH