WorldWideScience

Sample records for based recombinant hepatitis

  1. Immunogenicity and efficacy testing in chimpanzees of an oral hepatitis B vaccine based on live recombinant adenovirus.

    OpenAIRE

    Lubeck, M D; Davis, A R; Chengalvala, M; Natuk, R J; Morin, J E; Molnar-Kimber, K; Mason, B. B.; Bhat, B M; Mizutani, S; Hung, P P

    1989-01-01

    As a major cause of acute and chronic liver disease as well as hepatocellular carcinoma, hepatitis B virus (HBV) continues to pose significant health problems world-wide. Recombinant hepatitis B vaccines based on adenovirus vectors have been developed to address global needs for effective control of hepatitis B infection. Although considerable progress has been made in the construction of recombinant adenoviruses that express large amounts of HBV gene products, preclinical immunogenicity and ...

  2. Establishment of a simple assay in vitro for hepatitis C virus NS3 serine protease based on recombinant substrate and single—Chain protease

    Institute of Scientific and Technical Information of China (English)

    Rong-BinGuan; Yi-GangTong; Hai-TaoWang; Gui-XinDu; Li-HuaHou

    2002-01-01

    AIM:To establish a simple and convenient assay in vitro for the Hepatitis C virus NS3 serine prtease based on the recombinant protease and substrate,and to evaluate its feasibility in screening the enzyme inhibitors.

  3. Development of an infectious surrogate hepatitis C virus based on a recombinant vesicular stomatitis virus expressing hepatitis C virus envelope glycoproteins and green fluorescent protein.

    Science.gov (United States)

    Okuma, Kazu; Fukagawa, Koji; Tateyama, Seiji; Kohma, Takuya; Mochida, Keiko; Hiyoshi, Masateru; Takahama, Youichi; Hamaguchi, Yukio; Hirose, Kunitaka; Buonocore, Linda; Rose, John K; Mizuochi, Toshiaki; Hamaguchi, Isao

    2015-01-01

    To develop surrogate viruses for hepatitis C virus (HCV), we previously produced recombinant vesicular stomatitis viruses (rVSVs) lacking glycoprotein G but instead expressing chimeric HCV E1/E2 fused to G. These rVSVs were not infectious in HCV-susceptible hepatoma cells. In this study, to develop an infectious surrogate HCV based on an rVSV (vesicular stomatitis virus [VSV]/HCV), we generated a novel rVSV encoding the native E1/E2 (H77 strain) and green fluorescent protein (GFP) instead of G. Here, we showed that this VSV/HCV efficiently infected human hepatoma cells, including Huh7 human hepatoma cells, expressed GFP in these cells, and propagated, but did not do so in nonsusceptible BHK-21 cells. The infectivity of VSV/HCV, measured as the number of foci of GFP-positive cells, was specifically reduced by the addition of chimpanzee anti-HCV serum, anti-E2 antibody, or anti-CD81 antibody to the cultures. When sera obtained from HCV-infected or uninfected patients were added, infection was selectively inhibited only by the sera of HCV-infected patients. These data together suggest that this infectious GFP-expressing VSV/HCV could be a useful tool for studying the mechanisms of HCV entry into cells and for assessing potential inhibitors of viral entry, including neutralizing antibodies. PMID:25672345

  4. Epitope-based recombinant diagnostic antigen to distinguish natural infection from vaccination with hepatitis A virus vaccines.

    Science.gov (United States)

    Su, Qiudong; Guo, Minzhuo; Jia, Zhiyuan; Qiu, Feng; Lu, Xuexin; Gao, Yan; Meng, Qingling; Tian, Ruiguang; Bi, Shengli; Yi, Yao

    2016-07-01

    Hepatitis A virus (HAV) infection can stimulate the production of antibodies to structural and non-structural proteins of the virus. However, vaccination with an inactivated or attenuated HAV vaccine produces antibodies mainly against structural proteins, whereas no or very limited antibodies are produced against the non-structural proteins. Current diagnostic assays to determine exposure to HAV, such as the Abbott HAV AB test, detect antibodies only to the structural proteins and so are not able to distinguish a natural infection from vaccination with an inactivated or attenuated virus. Here, we constructed a recombinant tandem multi-epitope diagnostic antigen (designated 'H1') based on the immune-dominant epitopes of the non-structural proteins of HAV to distinguish the two situations. H1 protein expressed in Escherichia coli and purified by affinity and anion exchange chromatography was applied in a double-antigen sandwich ELISA for the detection of anti-non-structural HAV proteins, which was confirmed to distinguish a natural infection from vaccination with an inactivated or attenuated HAV vaccine. PMID:26994964

  5. Toolbox for Non-Intrusive Structural and Functional Analysis of Recombinant VLP Based Vaccines: A Case Study with Hepatitis B Vaccine

    OpenAIRE

    Mulder, Anke M.; Bridget Carragher; Victoria Towne; Yuan Meng; Yang Wang; Lance Dieter; Potter, Clinton S.; Washabaugh, Michael W.; Sitrin, Robert D; Qinjian Zhao

    2012-01-01

    BACKGROUND: Fundamental to vaccine development, manufacturing consistency, and product stability is an understanding of the vaccine structure-activity relationship. With the virus-like particle (VLP) approach for recombinant vaccines gaining popularity, there is growing demand for tools that define their key characteristics. We assessed a suite of non-intrusive VLP epitope structure and function characterization tools by application to the Hepatitis B surface antigen (rHBsAg) VLP-based vaccin...

  6. Development and validation of a genotype 3 recombinant protein-based immunoassay for hepatitis E virus serology in swine

    Directory of Open Access Journals (Sweden)

    W.H.M. van der Poel

    2014-04-01

    Full Text Available Hepatitis E virus (HEV is classified within the family Hepeviridae, genus Hepevirus. HEV genotype 3 (Gt3 infections are endemic in pigs in Western Europe and in North and South America and cause zoonotic infections in humans. Several serological assays to detect HEV antibodies in pigs have been developed, at first mainly based on HEV genotype 1 (Gt1 antigens. To develop a sensitive HEV Gt3 ELISA, a recombinant baculovirus expression product of HEV Gt3 open reading frame-2 was produced and coated onto polystyrene ELISA plates. After incubation of porcine sera, bound HEV antibodies were detected with anti-porcine anti-IgG and anti-IgM conjugates. For primary estimation of sensitivity and specificity of the assay, sets of sera were used from pigs experimentally infected with HEV Gt3. For further validation of the assay and to set the cutoff value, a batch of 1100 pig sera was used. All pig sera were tested using the developed HEV Gt3 assay and two other serologic assays based on HEV Gt1 antigens. Since there is no gold standard available for HEV antibody testing, further validation and a definite setting of the cutoff of the developed HEV Gt3 assay were performed using a statistical approach based on Bayes' theorem. The developed and validated HEV antibody assay showed effective detection of HEV-specific antibodies. This assay can contribute to an improved detection of HEV antibodies and enable more reliable estimates of the prevalence of HEV Gt3 in swine in different regions.

  7. Establishment of a simple assay in vitro for hepatitis C virus NS3 serine protease based on recombinant substrate and single-chain protease

    Institute of Scientific and Technical Information of China (English)

    Gui-Xin Du; Li-Hua Hou; Rong-Bin Guan; Yi-Gang Tong; Hai-Tao Wang

    2002-01-01

    AIM: To establish a simple and convenient assay in vitro for the Hepatitis C virus NS3 serine protease based on the recombinant protease and substrate, and to evaluate its feasibility in screening the enzyme inhibitors. METHODS: Based on the crystallographic structure of hepatitis C virus (HCV) serine protease, a novel single-chain serine protease was designed, in which the central sequence of cofactor NS4A was linked to the N-terminus of NS3 serine protease domain via a flexible linker GSGS. The fusion gene was obtained by two-step PCR that was carried out with three primers and then cloned into the prokaryotic expression vector pQE30, and the recombinant clone was verified by DNA sequencing. The single-chain recombinant protease was expressed when the E.coliwas induced with IPTG and the expression conditions were optimized to produce large amount of soluble protease. The recombinant substrate NS5ab that covers the cleavage point NS5A/B was also expressed in E.coli. Both of the protease and substrate were purified by using Ni-NTA agarose metal affinity resin, then they were mixed together in a specific buffer, and the mixture was analyzed by SDS-PAGE. The cleavage system was used to evaluate some compounds for their inhibitory activity on serine protease.RESULTS: The single-chain recombinant protease was overexpressed as soluble protein when the E. coliwas induced at a low dosage of IPTG (0.2 mM) and cultured at a low temperature (15℃). The protease was purified by using Ni-NTA agarose metal affinity resin (the purity is over 95 %).The recombinant substrate NS5ab was expressed in an insoluble form and could refold successfully after purification and dialysis. A simple and convenient assay in vitro was established, in which the purified single-chain serine protease could cleave the recombinant substrate NS5ab into two fragments that were visualized by SDS-PAGE. PMSF had an effect on inhibiting activity of serine protease, while EDTA had not.CONCLUSION: A simple

  8. Development of a Recombinant Cell-Based Indirect Immunofluorescence Assay for the Determination of Autoantibodies against Soluble Liver Antigen in Autoimmune Hepatitis

    Directory of Open Access Journals (Sweden)

    Christiane Radzimski

    2013-01-01

    Full Text Available Autoantibodies against soluble liver antigen (SLA are specific markers for autoimmune hepatitis (AIH type 1. In contrast to the determination of other AIH-associated autoantibodies by indirect immunofluorescence assay (IFA, detection of anti-SLA relied up to now on ELISA or immunoblot based on bacterially expressed recombinant protein. In order to develop a complementary IFA substrate, SLA isoform 1 was recombinantly produced in the human cell line HEK293 and controlled by a rabbit hyperimmune serum against SLA. The recombinant cells were used in IFA (RC-IFA to analyze sera from 20 AIH patients with anti-SLA positivity predetermined by ELISA together with 80 controls (20 anti-SLA negative AIH, 15 primary biliary cirrhosis, 15 HCV, and 30 healthy blood donors. Using RC-IFA, anti-SLA was detected in all ELISA positive AIH sera but in none of the controls. Furthermore, a cytosolic fraction of HEK293 containing SLA was able to neutralize the autoantibodies in all positive sera in a dose-dependent manner. HEK293 cells expressing SLA are a valid substrate for the serodiagnosis of AIH relevant autoantibodies by IFA. In concert with cryosections of primate liver, rat kidney, rat liver, rat stomach, and HEp-2 cells, they enable the parallel determination of all autoantibodies associated with autoimmune liver diseases.

  9. Serendipitous identification of natural intergenotypic recombinants of hepatitis C in Ireland.

    LENUS (Irish Health Repository)

    Moreau, Isabelle

    2006-01-01

    BACKGROUND: Recombination between hepatitis C single stranded RNA viruses is a rare event. Natural viable intragenotypic and intergenotypic recombinants between 1b-1a, 1a-1c and 2k-1b, 2i-6p, respectively, have been reported. Diagnostically recombinants represent an intriguing challenge. Hepatitis C genotype is defined by interrogation of the sequence composition of the 5\\' untranslated region [5\\'UTR]. Occasionally, ambiguous specimens require further investigation of the genome, usually by interrogation of the NS5B region. The original purpose of this study was to confirm the existence of a suspected mixed genotype infection of genotypes 2 and 4 by clonal analysis at the NS5B region of the genome in two specimens from two separate individuals. This initial identification of genotype was based on analysis of the 5\\'UTR of the genome by reverse line probe hybridisation [RLPH]. RESULTS: The original diagnosis of a mixed genotype infection was not confirmed by clonal analysis of the NS5B region of the genome. The phylogenetic analysis indicated that both specimens were natural intergenotypic recombinant forms of HCV. The recombination was between genotypes 2k and 1b for both specimens. The recombination break point was identified as occurring within the NS2 region of the genome. CONCLUSION: The viral recombinants identified here resemble the recombinant form originally identified in Russia. The RLPH pattern observed in this study may be a signature indicative of this particular type of intergenotype recombinant of hepatitis C meriting clonal analysis of NS2.

  10. Toolbox for non-intrusive structural and functional analysis of recombinant VLP based vaccines: a case study with hepatitis B vaccine.

    Directory of Open Access Journals (Sweden)

    Anke M Mulder

    Full Text Available BACKGROUND: Fundamental to vaccine development, manufacturing consistency, and product stability is an understanding of the vaccine structure-activity relationship. With the virus-like particle (VLP approach for recombinant vaccines gaining popularity, there is growing demand for tools that define their key characteristics. We assessed a suite of non-intrusive VLP epitope structure and function characterization tools by application to the Hepatitis B surface antigen (rHBsAg VLP-based vaccine. METHODOLOGY: The epitope-specific immune reactivity of rHBsAg epitopes to a given monoclonal antibody was monitored by surface plasmon resonance (SPR and quantitatively analyzed on rHBsAg VLPs in-solution or bound to adjuvant with a competitive enzyme-linked immunosorbent assay (ELISA. The structure of recombinant rHBsAg particles was examined by cryo transmission electron microscopy (cryoTEM and in-solution atomic force microscopy (AFM. PRINCIPAL FINDINGS: SPR and competitive ELISA determined relative antigenicity in solution, in real time, with rapid turn-around, and without the need of dissolving the particulate aluminum based adjuvant. These methods demonstrated the nature of the clinically relevant epitopes of HBsAg as being responsive to heat and/or redox treatment. In-solution AFM and cryoTEM determined vaccine particle size distribution, shape, and morphology. Redox-treated rHBsAg enabled 3D reconstruction from CryoTEM images--confirming the previously proposed octahedral structure and the established lipid-to-protein ratio of HBsAg particles. Results from these non-intrusive biophysical and immunochemical analyses coalesced into a comprehensive understanding of rHBsAg vaccine epitope structure and function that was important for assuring the desired epitope formation, determinants for vaccine potency, and particle stability during vaccine design, development, and manufacturing. SIGNIFICANCE: Together, the methods presented here comprise a novel

  11. Comparison of Two Recombinant Hepatitis B Vaccines

    OpenAIRE

    Hasan Nikui Nejad; Gholamali Ghorbani; Reza Razaghi; Hossain Akbari

    2009-01-01

    Background and Aims: Hepatitis B virus (HBV) infection is an important public health problem. Hepatitis B vaccine induces protective response in the majority of vaccinated persons. In our country, we do not have any evaluation for the efficacy of each type of vaccines used in adult and our objective was therefore to evaluate the efficacy of vaccines..Methods: In a randomized double-blind clinical trial 347 military personnel and their family in Kashan city, central Iran, were studied during A...

  12. Antibody responses to recombinant and plasma derived hepatitis B vaccines.

    OpenAIRE

    Brown, S E; Stanley, C.; Howard, C. R.; Zuckerman, A J; Steward, M W

    1986-01-01

    The antibody response to hepatitis B surface antigen (anti-HBs) induced in 25 recipients of a recombinant hepatitis B vaccine derived from yeast was compared with that induced in 25 recipients of a vaccine prepared from hepatitis B surface antigen (HBsAg) derived from plasma. Anti-HBs affinity and specificity were compared using assays of antibody affinity with two different antigens, a complex of the major polypeptide of HBsAg (p25; molecular weight 25 000 daltons) covalently linked to its g...

  13. Yeast-recombinant hepatitis B vaccine: efficacy with hepatitis B immune globulin in prevention of perinatal hepatitis B virus transmission

    International Nuclear Information System (INIS)

    A yeast-recombinant hepatitis B vaccine was licensed recently by the Food and Drug administration and is now available. To assess the efficacy of the yeast-recombinant vaccine, the authors administered the vaccine in combination with hepatitis B immune globulin to high-risk newborns. If infants whose mothers were positive for both hepatitis B surface antigen and the e antigen receive no immunoprophylaxis, 70% to 90% become infected with the virus, and almost all become chronic carriers. Among infants in this study who received hepatitis B immune globulin at birth and three 5-+g doses of yeast-recombinant hepatitis B vaccine, only 4.8% became chronic carriers, a better than 90% level of protection and a rate that is comparable with that seen with immune globulin and plasma-derived hepatitis B vaccine. Hepatitis surface antigen and antibodies were detected by radioimmunoassay. These data suggest that, in this high-risk setting, the yeast-recombinant vaccine is as effective as the plasma-derived vaccine in preventing hepatitis B virus infection and the chronic carrier state

  14. Bivalent inactivated hepatitis A and recombinant hepatitis B vaccine.

    Science.gov (United States)

    Beran, Jiri

    2007-12-01

    Hepatitis A and B remain serious global public health problems. Monovalent vaccines against hepatitis A and B have been available for many years. Since 1996, licenses have been gradually introduced for different formulations and immunization schedules of the first combined vaccines against both diseases. Twinrix Adult (with conventional and accelerated schedules) is available for the immunization of individuals aged 16 years or older in Europe and 18 years or older the USA. Twinrix Pediatric, with its three-dose schedule, and AmBirix, with its two-dose schedule, are licensed in Europe for ages 1-15 years. These vaccines offer a single injection for satisfactory protection against hepatitis A and B and an excellent safety and reactogenicity profile in comparison with monovalent vaccines. This article focuses on immunogenicity of the vaccines and proposes expert opinion and future directions in this field.

  15. Thrombocytopenic purpura as adverse reaction to recombinant hepatitis B vaccine

    OpenAIRE

    Ronchi, F; Cecchi, P; Falcioni, F.; Marsciani, A; Minak, G.; Muratori, G; Tazzari, P; Beverini, S

    1998-01-01

    Three cases of immune thrombocytopenic purpura after the first dose of recombinant hepatitis B vaccine occurred in infants under 6 months of age. Other possible causes of this condition were excluded. Antiplatelet antibodies were present. A defect in platelet production was excluded in two children. Corticosteroid treatment was effective. Subsequent administration of other vaccines (against polio, diphtheria, and tetanus) did not cause relapse of thrombocytopenia.



  16. A First Case of Hepatic Angiosarcoma Treated with Recombinant Interleukin-2

    OpenAIRE

    Mitsui, Fukiko; Aikata, Hiroshi; Hashimoto, Yoshimasa; Nagaoki, Yuko; KIMURA, Yuki; Katamura, Yoshio; Kawaoka, Tomokazu; Takaki, Shintaro; Hiraga, Nobuhiko; Tsuge, Masataka; Waki, Koji; Hiramatsu, Akira; Imamura, Michio; Kawakami, Yoshiiku; Takahashi, Shoichi

    2011-01-01

    A 60 year-old woman was admitted to our hospital because of management of multiple liver tumors. According to image findings and liver biopsy, she was diagnosed as having epithelioid hemangioendothelioma of the liver accompanied by metastases in the spleen, lungs and bones. Based on the spread of the liver tumors and the extensive systemic metastases, she was considered inoperable. Instead, she received hepatic arterial infusion therapy using recombinant interleukin-2. However, she died due t...

  17. Adaptive Mutations Enhance Assembly and Cell-to-Cell Transmission of a High-Titer Hepatitis C Virus Genotype 5a Core-NS2 JFH1-Based Recombinant

    DEFF Research Database (Denmark)

    Mathiesen, Christian K.; Prentoe, Jannick; Meredith, Luke W.;

    2015-01-01

    requiring high virus concentrations, such as studies of HCV particle composition and development of whole-virus vaccine antigens. IMPORTANCE: Hepatitis C virus (HCV) is a major global health care burden, affecting more than 150 million people worldwide. These individuals are at high risk of developing......UNLABELLED: Recombinant hepatitis C virus (HCV) clones propagated in human hepatoma cell cultures yield relatively low infectivity titers. Here, we adapted the JFH1-based Core-NS2 recombinant SA13/JFH1C3405G,A3696G (termed SA13/JFH1orig), of the poorly characterized genotype 5a, to Huh7.5 cells......, yielding a virus with greatly improved spread kinetics and an infectivity titer of 6.7 log10 focus-forming units (FFU)/ml. We identified several putative adaptive amino acid changes. In head-to-head infections at fixed multiplicities of infection, one SA13/JFH1orig mutant termed SA13/JFH1Core-NS5B...

  18. Vaccination against hepatitis B: comparison of intradermal and intramuscular administration of plasma derived and recombinant vaccines.

    OpenAIRE

    Payton, C. D.; Scarisbrick, D A; Sikotra, S.; Flower, A J

    1993-01-01

    A retrospective analysis of levels of antibody to hepatitis B surface antigen in 1419 health care workers was carried out to compare the efficacy of intramuscular and intradermal administration of plasma derived and recombinant hepatitis B vaccines. No significant difference was detected between the response to intradermal and intramuscular plasma derived vaccine. However of those who received intramuscular recombinant vaccine 81.6%, 13.8% and 4.7% were good (> or = 100 miu/ml), low (10-99 mi...

  19. Induced HBs antigenemia in healthy adults after immunization with two different hepatitis B recombinant vaccines

    OpenAIRE

    Masoud ZIAEE; Saádatjoo, Alireza; Mohamadpour, Malihe; Namaei, Mohammad Hasan

    2010-01-01

    Background and Aims Currently, vaccination is the most effective protective tool against hepatitis B virus infection. Some studies have shown that positive results for a hepatitis B virus surface antigen (HBsAg) test may be seen after vaccination. Materials and Methods In this clinical trial study, 62 healthy adult volunteers were randomly assigned to receive either the Engerix-B or the Hepavax-Gene hepatitis B recombinant vaccine. Blood samples were drawn 1, 3, and 5 days after vaccination a...

  20. Characterization of hepatitis C virus intergenotypic recombinant strains and associated virological response to sofosbuvir/ribavirin

    NARCIS (Netherlands)

    Hedskog, C.; Doehle, B.; Chodavarapu, K.; Gontcharova, V.; Crespo Garcia, J.; Knegt, R.; Drenth, J.P.H.; McHutchison, J.G.; Brainard, D.; Stamm, L.M.; Miller, M.D.; Svarovskaia, E.; Mo, H.

    2015-01-01

    To date, intergenotypic recombinant hepatitis C viruses (HCVs) and their treatment outcomes have not been well characterized. This study characterized 12 novel HCV recombinant strains and their response to sofosbuvir in combination with ribavirin (SOF/RBV) treatment. Across the phase II/III studies

  1. Comparative analysis of the molecular mechanisms of recombination in hepatitis C virus

    DEFF Research Database (Denmark)

    Galli, Andrea; Bukh, Jens

    2014-01-01

    Genetic recombination is an important evolutionary mechanism for RNA viruses. The significance of this phenomenon for hepatitis C virus (HCV) has recently become evident, with the identification of circulating recombinant forms in HCV-infected individuals and by novel data from studies permitted...... by advances in HCV cell culture systems and genotyping protocols. HCV is readily able to produce viable recombinants, using replicative and non-replicative molecular mechanisms. However, our knowledge of the required molecular mechanisms remains limited. Understanding how HCV recombines might be instrumental...... will focus on current data available on HCV recombination, also in relation to more detailed data from other RNA viruses....

  2. [Protection of health personnel against hepatitis B by DNA recombinant vaccine].

    Science.gov (United States)

    Navarrete-Navarro, S; Alvarez-Muñoz, M T; Bustamante-Calvillo, M E; Vallejo-Aguilar, O J; Muñoz, O; Santos-Preciado, J I; Avila-Figueroa, C

    1992-11-01

    Hepatitis B (HVB) is a worldwide spread health problem. It has been assessed that there are more than 300 millions of carriers. HVB has a special concern for health care workers (HCW's) due to the high risk among them of getting the infection in clinic-setting areas. According to some estimation, the risk for hepatitis B among HCW's is 2 to 10 times higher than the risk for general population. The risk is related to the degree of direct contact with blood and body fluids, as well as, with the frequency of traumatic exposure in the work place. The control of this infection is based on the observance of universal precautions and the vaccination, since there is not treatment against this disease. The results of an efficacy-evaluation of DNA recombinant vaccine against hepatitis B are reported; 174 HCW's were studied; three dosages of vaccine were administered (0.1st and 6th month) by I.M. via. In addition, three serum samples were collected at 0, 1st and 9th month after vaccine administration. We did not find carriers of surface antigen of hepatitis B. With regards to seroconverted individuals we observed the following results: there were a satisfactory response to the vaccine in 163 individuals (93.7%); however, 8 (4.6%) persons did not reach titles of protective antibodies and 3 (1.7%) did not show seroconversion at all. Therefore, 11 persons (6.3% of the total) did not result immunized. The secondary reactions to the vaccines were low in frequency and mainly of local presentation. Among the study population we did not find chronic carries of hepatitis B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1466772

  3. Productive homologous and non-homologous recombination of hepatitis C virus in cell culture.

    Directory of Open Access Journals (Sweden)

    Troels K H Scheel

    2013-03-01

    Full Text Available Genetic recombination is an important mechanism for increasing diversity of RNA viruses, and constitutes a viral escape mechanism to host immune responses and to treatment with antiviral compounds. Although rare, epidemiologically important hepatitis C virus (HCV recombinants have been reported. In addition, recombination is an important regulatory mechanism of cytopathogenicity for the related pestiviruses. Here we describe recombination of HCV RNA in cell culture leading to production of infectious virus. Initially, hepatoma cells were co-transfected with a replicating JFH1ΔE1E2 genome (genotype 2a lacking functional envelope genes and strain J6 (2a, which has functional envelope genes but does not replicate in culture. After an initial decrease in the number of HCV positive cells, infection spread after 13-36 days. Sequencing of recovered viruses revealed non-homologous recombinants with J6 sequence from the 5' end to the NS2-NS3 region followed by JFH1 sequence from Core to the 3' end. These recombinants carried duplicated sequence of up to 2400 nucleotides. HCV replication was not required for recombination, as recombinants were observed in most experiments even when two replication incompetent genomes were co-transfected. Reverse genetic studies verified the viability of representative recombinants. After serial passage, subsequent recombination events reducing or eliminating the duplicated region were observed for some but not all recombinants. Furthermore, we found that inter-genotypic recombination could occur, but at a lower frequency than intra-genotypic recombination. Productive recombination of attenuated HCV genomes depended on expression of all HCV proteins and tolerated duplicated sequence. In general, no strong site specificity was observed. Non-homologous recombination was observed in most cases, while few homologous events were identified. A better understanding of HCV recombination could help identification of natural

  4. Evaluation of immunogenicity and reactogenicity of recombinant DNA hepatitis B vaccine produced in India

    OpenAIRE

    Hussain, Zahid; Ali, Syed S.; Husain, Syed A.; Raish, Mohammad; Sharma, Deepika R; Kar, Premashis

    2005-01-01

    AIM: (1) To gain information on immune responses to an accelerated schedule of 0, 1, and 2 mo in paramedical staff and BDS students who are at an increased risk of getting hepatitis B infection and come under high risk groups. (2) To assess the efficacy and safety of Enivac-HB in different age groups, using genetically modified yeast strain Pichia pastoris, a new recombinant hepatitis B vaccine developed and manufactured in India.

  5. Multicenter study on the immunogenicity and safety of two recombinant vaccines against hepatitis B

    OpenAIRE

    Reinaldo de Menezes Martins; Gilberta Bensabath; Luiz Claudio Arraes; Maria de Lourdes Aguiar Oliveira; Juliana Custódio Miguel; Glayse Glayde Barbosa; Luiz Antonio Bastos Camacho

    2004-01-01

    The immunogenicity and safety of a new recombinant hepatitis B vaccine from the Instituto Butantan (Butang®) were evaluated in a multicenter, double-blind, prospective equivalence study in three centers in Brazil. Engerix B® was the standard vaccine. A total of 3937 subjects were recruited and 2754 (70%) met all protocol criteria at the end of the study. All the subjects were considered healthy and denied having received hepatitis B vaccine before the study. Study subjects who adhered to the ...

  6. Effectiveness of DNA-recombinant anti-hepatitis B vaccines in blood donors: a cohort study

    OpenAIRE

    Petry Andrea; de Souza Denise ER; Kupek Emil

    2007-01-01

    Abstract Background Although various studies have demonstrated efficacy of DNA-recombinant anti-hepatitis B vaccines, their effectiveness in health care settings has not been researched adequately. This gap is particularly visible for blood donors, a group of significant importance in the reduction of transfusion-transmitted hepatitis B. Methods This is a double cohort study of 1411 repeat blood donors during the period 1998–2002, involving a vaccinated and an unvaccinated cohort, with matchi...

  7. A novel complex A/C/G intergenotypic recombinant of hepatitis B virus isolated in southern China.

    Directory of Open Access Journals (Sweden)

    Heling Su

    Full Text Available Hepatitis B virus (HBV genotypes and subgenotypes may vary in geographical distribution and virological features. Previous investigations, including ours, showed that HBV genotypes B and C were respectively predominant in South and North China, while genotypes A and D were infrequently detected and genotype G was not found. In this study, a novel A/C/G intergenotype was identified in patients with chronic HBV infection in Guilin, a city in southern China. Initial phylogenetic analysis based on the S gene suggested the HBV recombinant to be genotype G. However, extended genotyping based on the entire HBV genome indicated it to be an A/C/G intergenotype with a closer relation to genotype C. Breakpoint analysis using the SIMPLOT program revealed that the recombinant had a recombination with a arrangement of genotypes A, G, A and C fragments. Compared with the HBV recombinants harboring one or two genotype G fragments found in Asian countries, this Guilin recombinant was highly similar to the Vietnam (98-99% and Long An recombinants (96-99%, but had a relatively low similarity to the Thailand one (89%. Unlike those with the typical genotype G of HBV, the patients with the Guilin recombinant were seropositive for HBeAg. Moreover, a relatively high HBV DNA viral load (>2 × 10(6 IU/ml was detected in the patients, and the analysis of viral replication capacity showed that the Guilin recombinant strains had a competent replication capacity similar to genotypes B and C strains. These findings can aid in not only the clarification of the phylogenetic origin of the HBV recombinants with the genotype G fragment found in Asian countries, but also the understanding of the virological properties of these complicated HBV recombinants.

  8. Recombinant Hepatitis A Virus Antigen: Improved Production and Utility in Diagnostic Immunoassays

    OpenAIRE

    LaBrecque, F. D.; LaBrecque, D. R.; Klinzman, D.; Perlman, S; Cederna, J. B.; Winokur, P. L.; Han, J.-Q.; Stapleton, J. T.

    1998-01-01

    Hepatitis A virus (HAV) immunoassays use cell culture-derived HAV antigen to detect HAV-specific antibodies. The current method of production of HAV antigen in tissue culture is time-consuming and expensive. We previously expressed the HAV open reading frame in recombinant vaccinia viruses (rV-ORF). The recombinant HAV polyprotein was accurately processed and was assembled into subviral particles. These particles were bound by HAV-neutralizing antibodies and were able to elicit antibodies whi...

  9. The role of recombinant IL-12 in enhancing immune responses induced by hepatitis B vaccine in mice

    International Nuclear Information System (INIS)

    Objective: To study the role played by recombinant IL-12 in enhancing the intensity and quality of the immune response to hepatitis B vaccine in mice, and investigate the possibility of adding recombinant IL-12 as adjuvants to hepatitis B therapeutic vaccine. Methods: Recombinant IL-12 was injected together with hepatitis B vaccine into mice and special anti-HBsAb in the mice and the cellular immune responses were examined. Results: Recombinant IL-12 can obviously enhance T lymphocyte multiplication activity, accelerate excretion of cytokines IFN-γ and IL-2, and increase the IgG2a antibody in mice. Conclusion: Recombinant IL-12 can remarkably strengthen the cellular immune responses induced by the hepatitis B vaccine, and modulate the immune responses toward Thl. (authors)

  10. Comparison of the effect of two different doses of recombinant hepatitis B vaccine on immunogenicity in healthy adults

    OpenAIRE

    Li, Jing; Yao, Jun; Shan, Huan; Chen, Yongdi; Jiang, Zheng-gang; Ren, Jing-jing; Xu, Kai-Jin; Ruan, Bing; Yang, Shi-gui; Wang, Bing; Xie, Tian-sheng; Li, Qian

    2015-01-01

    The aim of this study was to evaluate the one-month immune response to 2 different doses (10 and 20 μg) of recombinant hepatitis B vaccine in adults aged 20–46 y. Subjects who were negative for hepatitis B surface antigen (HBsAg), hepatitis B antibody (anti-HBs), and hepatitis B core antibody (anti-HBc) were recruited. The participants were divided into 2 groups: group I received 3 doses of 10 μg hepatitis B vaccine at 0, 1 and 3 months, and group II received 3 doses of 20 μg at the same time...

  11. Hepatitis B vaccines prepared from yeast by recombinant DNA techniques: Memorandum from a WHO Meeting

    OpenAIRE

    1985-01-01

    A meeting of experts was convened on 19-21 November 1984 in Geneva, Switzerland, to advise the World Health Organization on the production of hepatitis B vaccine prepared from yeast (Saccharomyces cerevisiae), using recombinant DNA technology. This vaccine development follows the advances in molecular genetics that have permitted genes coding for biologically active substances to be identified, analysed with fine precision, transferred within and between host organisms, and expressed under co...

  12. Evidence of recombination in Hepatitis C Virus populations infecting a hemophiliac patient

    Directory of Open Access Journals (Sweden)

    Cristina Juan

    2009-11-01

    Full Text Available Abstract Background/Aim Hepatitis C virus (HCV infection is an important cause of morbidity and mortality in patients affected by hereditary bleeding disorders. HCV, as others RNA virus, exploit all possible mechanisms of genetic variation to ensure their survival, such as recombination and mutation. In order to gain insight into the genetic variability of HCV virus strains circulating in hemophiliac patients, we have performed a phylogenetic analysis of HCV strains isolated from 10 patients with this kind of pathology. Methods Putative recombinant sequence was identified with the use of GARD program. Statistical support for the presence of a recombination event was done by the use of LARD program. Results A new intragenotypic recombinant strain (1b/1a was detected in 1 out of the 10 hemophiliac patient studied. The recombination event was located at position 387 of the HCV genome (relative to strain AF009606, sub-type 1a corresponding to the core gene region. Conclusion Although recombination may not appear to be common among natural populations of HCV it should be considered as a possible mechanism for generating genetic diversity in hemophiliacs patients.

  13. Algae-based oral recombinant vaccines

    OpenAIRE

    Specht, Elizabeth A.; Mayfield, Stephen P

    2014-01-01

    Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for “molecular pharming” in food crops has waned in the last decade due to difficulty in ...

  14. Response of booster dose of cuban recombinant hepatitis-B vaccine in nonresponder and hyporesponder children

    International Nuclear Information System (INIS)

    Acute hepatitis B infection can debilitate a patient for weeks and occasionally has a fatal outcome, while chronic infection is a major threat to the individual. To assess response of nonresponder and hyporesponder children to booster dose of Cuban recombinant hepatitis B vaccine. An interventional, descriptive study has been conducted on children who had been immunized with Cuban recombinant Hepatitis B vaccine and their antibody titers were <10mIU/ml (nonresponder) and 10-100mIU/ml (hyporesponder) administered booster dose of the same vaccine in their Deltoid muscles. The response of 141 children with the mean age of 1.9 years to booster dose of vaccine were 94.3% and 100% vaccines with the first and second booster dose of vaccination respectively. The anti-HBs titer in nonresponders and hyporesponders were 468+-346 and 783+-346mIU/ml respectively with significant differences between two groups (P=0.001). This study demonstrate moderately increase antibody production in the majority of vaccines with single supplementary vaccine. (author)

  15. The Last Ten Years of Advancements in Plant-Derived Recombinant Vaccines against Hepatitis B

    Science.gov (United States)

    Joung, Young Hee; Park, Se Hee; Moon, Ki-Beom; Jeon, Jae-Heung; Cho, Hye-Sun; Kim, Hyun-Soon

    2016-01-01

    Disease prevention through vaccination is considered to be the greatest contribution to public health over the past century. Every year more than 100 million children are vaccinated with the standard World Health Organization (WHO)-recommended vaccines including hepatitis B (HepB). HepB is the most serious type of liver infection caused by the hepatitis B virus (HBV), however, it can be prevented by currently available recombinant vaccine, which has an excellent record of safety and effectiveness. To date, recombinant vaccines are produced in many systems of bacteria, yeast, insect, and mammalian and plant cells. Among these platforms, the use of plant cells has received considerable attention in terms of intrinsic safety, scalability, and appropriate modification of target proteins. Research groups worldwide have attempted to develop more efficacious plant-derived vaccines for over 30 diseases, most frequently HepB and influenza. More inspiring, approximately 12 plant-made antigens have already been tested in clinical trials, with successful outcomes. In this study, the latest information from the last 10 years on plant-derived antigens, especially hepatitis B surface antigen, approaches are reviewed and breakthroughs regarding the weak points are also discussed. PMID:27754367

  16. A universal BMV-based RNA recombination system--how to search for general rules in RNA recombination.

    Science.gov (United States)

    Alejska, Magdalena; Figlerowicz, Magdalena; Malinowska, Nelli; Urbanowicz, Anna; Figlerowicz, Marek

    2005-07-07

    At present, there is no doubt that RNA recombination is one of the major factors responsible for the generation of new RNA viruses and retroviruses. Numerous experimental systems have been created to investigate this complex phenomenon. Consequently, specific RNA structural motifs mediating recombination have been identified in several viruses. Unfortunately, up till now a unified model of genetic RNA recombination has not been formulated, mainly due to difficulties with the direct comparison of data obtained for different RNA-based viruses. To solve this problem, we have attempted to construct a universal system in which the recombination activity of various RNA sequences could be tested. To this end, we have used brome mosaic virus, a model (+)RNA virus of plants, for which the structural requirements of RNA recombination are well defined. The effectiveness of the new homomolecular system has been proven in an experiment involving two RNA sequences derived from the hepatitis C virus genome. In addition, comparison of the data obtained with the homomolecular system with those generated earlier using the heteromolecular one has provided new evidence that the mechanisms of homologous and non-homologous recombination are different and depend on the virus' mode of replication.

  17. Thyroid dysfunction induced by recombinant interferon-alpha therapy for chronic active type C hepatitis

    International Nuclear Information System (INIS)

    The aim of this study was to assess the frequency and types of thyroid therapy in patients of Chronic dysfunction that develops during IFN- Hepatitis C. The study was carried out on a total of 50 patients of chronic therapy. In addition 50 patients with hepatitis C on recombinant IFN- chronic hepatitis C, not on any antiviral treatment, were included as controls. After informed consent, clinical history was obtained, physical examination was done and findings recorded on a pre-designed proforma. Blood sampling was done for thyroid profile at the beginning of interferon therapy, at 12 weeks and finally at 24 weeks. Thyroid dysfunction (TD) was observed in 14% (n=7) of the patients on antiviral therapy for CHC (n=50). Amongst these seven patients with TD, hypothyroidism was observed in 5 and hyperthyroidism in 2 patients. In contrast the frequency of thyroid dysfunction observed in control group (n=50) was 2%. The frequency of thyroid dysfunction in patients of chronic hepatitis C treated with interferon approaches 14%, with hypothyroidism being the more commonly observed pattern. (author)

  18. [Characterization of a panel of monoclonal antibodies to hepatitis C NS3 recombinant protein ].

    Science.gov (United States)

    Abdulmedzhidova, A G; Masalova, O V; Atanadze, S N; Ulanova, T I; Burkov, A N; Khudiakov, Iu E; Fields, H; Kushch, A A

    2002-01-01

    Recombinant protein rNS3 imitating helicase region (1356-1459 amino acid residues) of hepatitis C virus (HCV) was expressed in E. coli cells and used for BALB/c mice immunization. Seven hybrydoma clones producing monoclonal antibodies (MAbs) to rHS3 were obtained. All MAbs reacted in ELISA with NS3 protein from Murex anti-HCV Version III and in immunoblotting from RIBA 3. These MAbs detect 5 individual epitopes, 4 of which were conformational and 1 discontinuous. All MAbs could compete for rNS3 binding with serum antibodies from patients with chronic hepatitis C, which suggests that these MAbs can recognize the natural HCV NS3 protein.

  19. Effectiveness of DNA-recombinant anti-hepatitis B vaccines in blood donors: a cohort study

    Directory of Open Access Journals (Sweden)

    Petry Andrea

    2007-11-01

    Full Text Available Abstract Background Although various studies have demonstrated efficacy of DNA-recombinant anti-hepatitis B vaccines, their effectiveness in health care settings has not been researched adequately. This gap is particularly visible for blood donors, a group of significant importance in the reduction of transfusion-transmitted hepatitis B. Methods This is a double cohort study of 1411 repeat blood donors during the period 1998–2002, involving a vaccinated and an unvaccinated cohort, with matching of the two in terms of sex, age and residence. Average follow-up was 3.17 person-years. The outcome measure was infection with hepatitis B virus (HBV, defined by testing positive on serologic markers HBsAg or anti-HBC. All blood donors were from the blood bank in Joaçaba, federal state of Santa Catarina, Brazil. Results The cohorts did not differ significantly regarding sex, age and marital status but the vaccinated cohort had higher mean number of blood donations and higher proportion of those residing in the county capital Joaçaba. Hepatitis B incidences per 1000 person-years were zero among vaccinated and 2,33 among non-vaccinated, resulting in 100% vaccine effectiveness with 95% confidence interval from 30,1% to 100%. The number of vaccinated persons necessary to avoid one HBV infection in blood donors was estimated at 429 with 95% confidence interval from 217 to 21422. Conclusion The results showed very high effectiveness of DNA-recombinant anti-HBV vaccines in blood donors. Its considerable variation in this study is likely due to the limited follow-up and the influence of confounding factors normally balanced out in efficacy clinical trials.

  20. Effects of recombinant human adenovirus-p53 on the regression of hepatic fibrosis

    Science.gov (United States)

    Liu, Yehong; Yang, Puye; Chen, Na; Lin, Shumei; Liu, Min

    2016-01-01

    Hepatic fibrosis is a scarring process that may progress to hepatic cirrhosis and even hepatic carcinoma if left untreated. Hepatic stellate cells (HSCs) play essential roles in the development of hepatic fibrosis. The tumor suppressor protein p53 is a transcription factor that is involved in cell proliferation, cell cycle regulation, apoptosis and DNA repair. Recombinant human adenovirus-p53 (Ad-p53) has been demonstrated to act as a promising antitumor gene therapy in various types of cancer. However, there is limited infomration regarding the therapeutic effect of Ad-p53 on the regression of hepatic fibrosis. In order to examine the underlying molecular mechanism responsible for the effects of Ad-p53 on HSCs, a rat model of hepatic fibrosis was established and HSC-T6 cells were cultured under different conditions. The expression of p53, transforming growth factor (TGF-β1) and α-smooth muscle actin (α-SMA), which is a marker of activated HSCs, was detected by immunohistochemical assays and RT-qPCR. In vitro, five different concentrations (1×106, 5×106, 1×107, 2×107 and 5×107 PFU/ml) of Ad-p53 were selected for use in the MTT assay to analyze the proliferation of HSCs at 0, 24, 48 and 72 h. Flow cytometric analysis was applied to determine the effect of three different concentrations of Ad-p53 (5×106, 1×107 and 2×107 PFU/ml) on the cell cycle and the apoptosis of HSC-T6 cells at 24 and 48 h. The results of immunohistochemical studies and RT-qPCR showed that Ad-p53 upregulated the expression of p53, and downregulated the expression of TGF-β1 and α-SMA. The MTT assay revealed that when treated with various doses of Ad-p53, the proliferation of HSCs was inhibited within a certain range of concentrations and time periods. Analysis of flow cytometric data showed that Ad-p53 arrested the cell cycle in G1 phase and significantly induced apoptosis. Taken together, these findings suggest that Ad-p53 promotes apoptosis and inhibits the proliferation of HSCs in

  1. Algae-based oral recombinant vaccines.

    Science.gov (United States)

    Specht, Elizabeth A; Mayfield, Stephen P

    2014-01-01

    Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for "molecular pharming" in food crops has waned in the last decade due to difficulty in developing transgenic crop plants and concerns of contaminating the food supply. Microalgae could be poised to become the next candidate in recombinant subunit vaccine production, as they present several advantages over terrestrial crop plant-based platforms including scalable and contained growth, rapid transformation, easily obtained stable cell lines, and consistent transgene expression levels. Algae have been shown to accumulate and properly fold several vaccine antigens, and efforts are underway to create recombinant algal fusion proteins that can enhance antigenicity for effective orally delivered vaccines. These approaches have the potential to revolutionize the way subunit vaccines are made and delivered - from costly parenteral administration of purified protein, to an inexpensive oral algae tablet with effective mucosal and systemic immune reactivity. PMID:24596570

  2. Algae-based oral recombinant vaccines.

    Science.gov (United States)

    Specht, Elizabeth A; Mayfield, Stephen P

    2014-01-01

    Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for "molecular pharming" in food crops has waned in the last decade due to difficulty in developing transgenic crop plants and concerns of contaminating the food supply. Microalgae could be poised to become the next candidate in recombinant subunit vaccine production, as they present several advantages over terrestrial crop plant-based platforms including scalable and contained growth, rapid transformation, easily obtained stable cell lines, and consistent transgene expression levels. Algae have been shown to accumulate and properly fold several vaccine antigens, and efforts are underway to create recombinant algal fusion proteins that can enhance antigenicity for effective orally delivered vaccines. These approaches have the potential to revolutionize the way subunit vaccines are made and delivered - from costly parenteral administration of purified protein, to an inexpensive oral algae tablet with effective mucosal and systemic immune reactivity.

  3. Algae-based oral recombinant vaccines

    Directory of Open Access Journals (Sweden)

    Elizabeth A Specht

    2014-02-01

    Full Text Available Recombinant subunit vaccines are some of the safest and most effective vaccines available, but their high cost and the requirement of advanced medical infrastructure for administration make them impractical for many developing world diseases. Plant-based vaccines have shifted that paradigm by paving the way for recombinant vaccine production at agricultural scale using an edible host. However, enthusiasm for molecular pharming in food crops has waned in the last decade due to difficulty in developing transgenic crop plants and concerns of contaminating the food supply. Microalgae are poised to become the next candidate in recombinant subunit vaccine production, and they present several advantages over terrestrial crop plant-based platforms including scalable and contained growth, rapid transformation, easily obtained stable cell lines, and consistent transgene expression levels. Algae have been shown to accumulate and properly fold several vaccine antigens, and efforts are underway to create recombinant algal fusion proteins that can enhance antigenicity for effective orally-delivered vaccines. These approaches have the potential to revolutionize the way subunit vaccines are made and delivered – from costly parenteral administration of purified protein, to an inexpensive oral algae tablet with effective mucosal and system immune reactivity.

  4. A Quick and Simple Polarographic Method for Aluminum Measurement in Recombinant Hepatitis B Vaccine

    Directory of Open Access Journals (Sweden)

    Mahmoud Alebouyeh (PhD

    2016-01-01

    Full Text Available Background and Objective: Aluminum salts are among the most common useful additive compounds in preparation of human and animal vaccines. Aluminum phosphate and aluminum hydroxide are two additives that show good immunoadjuvant effects with many antigens. Aluminum-containing vaccines lead to a better and longer immune response compared to adjuvant-lacking vaccines. The Chromogenic methods used for determination of aluminum amounts in manufacturing centers are time-consuming and requires some experienced technicians to obtain accurate results. This study aimed to design and validate a simple polarographic method to measure aluminum in recombinant hepatitis B vaccine. Methods: In this study, the effects of temperature, pH, potential range and potential scan rate on the polarographic method of measuring aluminum in hepatitis B vaccine was evaluated and the optimal values for each of these factors were achieved. Results: In order to measure aluminum, temperature of 60 °C and pH of 4.5 were found as the optimal values. Implementation of polarographic method in the potential range of -0.25 to 0.1 volts had a better signal. Conclusion: Since the polarography method is more simple, accurate and faster than the chromogenic methods, it is suitable to be used for the measurement of aluminum in hepatitis B vaccine and it is recommended to be used in quality control laboratories for biological products.

  5. Crystallization and preliminary X-ray diffraction analysis of recombinant hepatitis E virus-like particle

    International Nuclear Information System (INIS)

    A recombinant virus-like particle that is a potential oral hepatitis E vaccine was crystallized. Diffraction data were collected to 8.3 Å resolution and the X-ray structure was phased with the aid of a low-resolution density map determined using cryo-electron microscopy data. Hepatitis E virus (HEV) accounts for the majority of enterically transmitted hepatitis infections worldwide. Currently, there is no specific treatment for or vaccine against HEV. The major structural protein is derived from open reading frame (ORF) 2 of the viral genome. A potential oral vaccine is provided by the virus-like particles formed by a protein construct of partial ORF3 protein (residue 70–123) fused to the N-terminus of the ORF2 protein (residues 112–608). Single crystals obtained by the hanging-drop vapour-diffusion method at 293 K diffract X-rays to 8.3 Å resolution. The crystals belong to space group P212121, with unit-cell parameters a = 337, b = 343, c = 346 Å, α = β = γ = 90°, and contain one particle per asymmetric unit

  6. Crystallization and preliminary X-ray diffraction analysis of recombinant hepatitis E virus-like particle

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Che-Yen [Molecular and Cellular Biology, University of California, Davis, CA 95616 (United States); Karolinska Institute Structural Virology, F68 Karolinska University Hospital, SE-14186 Stockholm (Sweden); Institute of Public Health, National Yang-Ming University, 112 Taipei,Taiwan (China); Miyazaki, Naoyuki [Molecular and Cellular Biology, University of California, Davis, CA 95616 (United States); Karolinska Institute Structural Virology, F68 Karolinska University Hospital, SE-14186 Stockholm (Sweden); Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Yamashita, Tetsuo [Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Institute for Microbial Diseases, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Higashiura, Akifumi; Nakagawa, Atsushi [Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Li, Tian-Cheng; Takeda, Naokazu [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Xing, Li [Molecular and Cellular Biology, University of California, Davis, CA 95616 (United States); Karolinska Institute Structural Virology, F68 Karolinska University Hospital, SE-14186 Stockholm (Sweden); Hjalmarsson, Erik; Friberg, Claes [Crystal Research AB, 22370 Lund (Sweden); Liou, Der-Ming [Institute of Public Health, National Yang-Ming University, 112 Taipei,Taiwan (China); Sung, Yen-Jen [Institute of Public Health, National Yang-Ming University, 112 Taipei,Taiwan (China); Institute of Anatomy and Cell Biology, National Yang-Ming University, 112 Taipei,Taiwan (China); Tsukihara, Tomitake [Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Matsuura, Yoshiharu [Institute for Microbial Diseases, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Miyamura, Tatsuo [Department of Virology II, National Institute of Infectious Diseases, Tokyo (Japan); Cheng, R. Holland, E-mail: rhch@ucdavis.edu [Molecular and Cellular Biology, University of California, Davis, CA 95616 (United States); Karolinska Institute Structural Virology, F68 Karolinska University Hospital, SE-14186 Stockholm (Sweden)

    2008-04-01

    A recombinant virus-like particle that is a potential oral hepatitis E vaccine was crystallized. Diffraction data were collected to 8.3 Å resolution and the X-ray structure was phased with the aid of a low-resolution density map determined using cryo-electron microscopy data. Hepatitis E virus (HEV) accounts for the majority of enterically transmitted hepatitis infections worldwide. Currently, there is no specific treatment for or vaccine against HEV. The major structural protein is derived from open reading frame (ORF) 2 of the viral genome. A potential oral vaccine is provided by the virus-like particles formed by a protein construct of partial ORF3 protein (residue 70–123) fused to the N-terminus of the ORF2 protein (residues 112–608). Single crystals obtained by the hanging-drop vapour-diffusion method at 293 K diffract X-rays to 8.3 Å resolution. The crystals belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 337, b = 343, c = 346 Å, α = β = γ = 90°, and contain one particle per asymmetric unit.

  7. Evaluation of immunogenicity and reactogenicity of recombinant DNA hepatitis B vaccine produced in India

    Institute of Scientific and Technical Information of China (English)

    Zahid Hussain; Syed S Ali; Syed A Husain; Mohammad Raish; Deepika R Sharma; Premashis Kar

    2005-01-01

    AIM: (1) To gain information on immune responses to an accelerated schedule of 0, 1, and 2 mo in paramedical staff and BDS students who are at an increased risk of getting hepatitis B infection and come under high risk groups. (2) To assess the efficacy and safety of EnivacHB in different age groups, using genetically modified yeast strain Pichia pastoris, a new recombinant hepatitis B vaccine developed and manufactured in India.METHODS: A prospective, comparative, and single blinded trial of rapid (0, 1, and 2 mo) hepatitis B immunization schedulewas reported. A total of three hundred and seven (212 females and 95 males) healthy volunteers divided into three age groups (18-29, 30-39,and 40-49) were enrolled after screening for markers of hepatitis B. All the volunteers received 20 mg of the vaccine intramuscularly at 0, 1, and 2 mo.RESULTS: Geometric mean titers were calculated pre and post vaccination. Before immunization the GMT was 0.0124 mIU/mL. One month after the administration of the third dose of recombinant vaccine 296/307 (96.5%)subjects achieved seroprotective levels of anti-HBs. The geometric mean anti-HBs titers achieved after one month of the third dose was 2 560.0 mIU/mL. The geometric mean anti-HBs titer of males was 2 029.0 mIU/mL, while that of the females was 2 759.0 mIU/mL. In the age group of 18-29 years, anti-HBs titer was 3 025.0 mIU/mL, while that in the age group of 30-39 years was 2096.0 mIU/mL. In third age group of 40-49 years, antiHBs titer was 1 592.0 mIU/mL. Hyper-responses (antiHBs≥100 mIU/mL) were shown in 88.0% (271/307) of subjects. Eleven (3.5%) subjects responded poorly to the vaccine in the age group of 40-49 years. There was only mild pain at the site of injection otherwise there were no other adverse drug reactions (ADRs).CONCLUSION: This vaccine (Enivac-HB) is safe and efficacious, providing significant protection after the third dose and rapid hepatitis B immunization schedule of 0, 1, and 2 mo can be recommended

  8. Construction of Recombinant Modified Vaccinia Ankara (MVA) Expressing Hepatitis B Virus Surface Antigen

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The T lymphocyte response has been shown to be the determinant in the clearance of many viral infections.Hence, therapeutic vaccine candidates against HBV are designed to enhance this response of the immune system.Vaccinia virus vector-based vaccines have been proposed as excellent candidates to elicit long-term and strong T lymphocyte mediated immune responses. In this study, the recombinant MVA expressing HBV surface antigen has been constructed, which can elicit a potent T cell mediated response. The ELISA results for the surface protein in the medium of the recombinant MVA, strongly indicate that the recombinant virus has been successfully obtained.

  9. Anti-inflammatory effect of recombinant thrombomodulin for fulminant hepatic failure.

    Science.gov (United States)

    Kurokohchi, Kazutaka; Imataki, Osamu; Kubo, Fumiyoshi

    2015-07-14

    Fulminant hepatic failure (FHF) is a critical illness that can be comorbid to primary liver damage. FHF shows a high mortality rate, and patients with FHF require intensive therapy, including plasma apheresis. However, intensive care at the present is not enough to restore the severe liver damage or promote hepatocellular reproduction, and a standard therapy for the treatment of FHF has not been established. An 86-year-old female with FHF was admitted to our hospital. Her manifestation demonstrated a clinical situation of systemic inflammatory response syndrome (SIRS) and disseminated intravascular coagulation. A diagnosis of fulminant hepatitis was made according to the definition given in the position paper of the American Association for the Study of Liver Diseases. Her serum hepatocyte growth factor (HGF) level had increased to 11.84 ng/mL. The HGF level indicated massive liver damage as seen in FHF. Recombinant thrombomodulin (rTM) was administered daily from the admission day for 1 wk at 380 U/kg. The patient's white blood cells and C-reactive protein responded to the rTM treatment within a few days. The HGF level and PT recovered to the normal range. The levels of proinflammatory cytokines (tumor necrosis factor-α and interleukin-1β) were suppressed by the administration of rTM. The patient's hepatic function (e.g., PT and albumin) completely recovered without plasma exchange. rTM may modulate the over-response of SIRS with the improvement of proinflammatory cytokines. The underlying mechanism is thought to be the inhibitory effect of rTM on high-mobility group box 1 (HMBG1). The pathogenesis of HMBG1 protein in fulminant hepatic failure has been already known. A novel favorable effect of rTM for SIRS would be promising for FHF, and the wide application of rTM for SIRS should be considered.

  10. Aluminum phosphate shows more adjuvanticity than Aluminum hydroxide in recombinant hepatitis –B vaccine formulation

    Directory of Open Access Journals (Sweden)

    2008-08-01

    Full Text Available Background: Although a number of investigation have been carried out to find alternative adjuvants to aluminum salts in vaccine formulations, they are still extensively used due to their good track record of safety, low cost and proper adjuvanticity with a variety of antigens. Adsorption of antigens onto aluminum compounds depends heavily on electrostatic forces between adjuvant and antigen. Commercial recombinant protein hepatitis B vaccines containing aluminum hydroxide as adjuvant is facing low induction of immunity in some sections of the vaccinated population. To follow the current global efforts in finding more potent hepatitis B vaccine formulation, adjuvanticity of aluminum phosphate has been compared to aluminum hydroxide. Materials and methods: The adjuvant properties of aluminum hydroxide and aluminum phosphate in a vaccine formulation containing a locally manufactured hepatitis B (HBs surface antigen was evaluated in Balb/C mice. The formulations were administered intra peritoneally (i.p. and the titers of antibody which was induced after 28 days were determined using ELISA technique. The geometric mean of antibody titer (GMT, seroconversion and seroprotection rates, ED50 and relative potency of different formulations were determined. Results: All the adjuvanicity markers obtained in aluminum phosphate formulation were significantly higher than aluminum hydroxide. The geometric mean of antibody titer of aluminum phosphate was approximately three folds more than aluminum hydroxide. Conclusion: Aluminum phosphate showed more adjuvanticity than aluminum hydroxide in hepatitis B vaccine. Therefore the use of aluminum phosphate as adjuvant in this vaccine may lead to higher immunity with longer duration of effects in vaccinated groups.

  11. Immunogenicity and reactogenicity of a recombinant hepatitis B vaccine in subjects over age of forty years and response of a booster dose among nonresponders

    Institute of Scientific and Technical Information of China (English)

    Kunal Das; R. K. Gupta; V. Kumar, P.Kar

    2003-01-01

    AIM: The study was initiated to evaluate the reactogenicity and immunogenicity of a recombinant hepatitis B vaccine in age group >40 years and to study the response of a single booster dose in primary non-responders to the hepatitis B vaccination.METHODS: A total of 102 volunteers without markers of hepatitis B infection (negative for HBsAg, anti-HBc antibody,HBeAg and anti-HBs antibody) received 20 pg of recombinant HB vaccine intramuscularly at 0, 1, and 6 months. Anti HBs titers were evaluated by a quantitative Elisa kit at 90 and 210 days. A booster dose of 20 pg HB vaccine was given after 6 months of the 3rd vaccine dose to the 15 nonresponders and anti-HBs titers were measured after 1 month.RESULTS: Seroprotection (anti-HBs GMT3 10 IU/L) was achieved in 85.3 % (87/102) volunteers. The mean GMT titers of the vaccine responders was 136.1 IU/L. Of the seroprotected individuals, there were 32.4 % (33/102) hyporesponders (antiHBs titers <10-99 mlU/ml) and 52.9 % (54/102) were responders (anti-HBs titers >100 IU/L). All the non-responders (15/15) responded to a single dose of the booster dose of recombinant HB vaccine and their mean anti-HBs antibody titers were more than 100.5 mIU/ml after the booster dose.CONCLUSION: Recombinant hepatitis B vaccine offers good seroprotection in the age group >40 years and has a good safety profile. A single booster dose after 6 months in primary non-responders leads to good seroprotective anti-HBs antibody titers. However, larger population based studies are needed to evaluate the role of a booster dose in selected group of non-responders and whether such an approach will be cost effective.

  12. High-Level Production of a Functional Recombinant Hepatitis B Virus Polymerase in Insect Cells with a Baculovirus Expression System

    Institute of Scientific and Technical Information of China (English)

    WANG Xiaoyan; GAO Linlin; DENG Fei; ZHANG Yanfang; LI Yan; LIN Jusheng

    2007-01-01

    HBV polymerase has intrinsic RNA-dependent reverse transcriptase, DNA-dependent DNA polymerase as well as RNaseH activity. Analysis of HBV polymerase has been hampered for many years due to the inability to express functional enzyme in a recombinant system. To obtain active polymerase at a high level, we have taken advantage of baculovirus expression system. The gene of HBV polymerase was amplified by PCR and cloned into pFastBac Dual to construct the recombinant plasmid pFastbac Dual-pol. The recombinant donor plasmid, pFastbac Dual-pol, was constructed by inserting HBV polymerase gene into EcoRI and PstI sites controlled by polyhedrin promoter. The recombinant donor plasmid was transformed into DH10Bac competent cells for transposition. Recombinant bacmid was constructed by inserting of the mini-Tn7 element from the donor plasmid into the mini-attTn7 attachment site on the bacmid. The recombinant bacmid DNA was isolated and transfected into the Sf9 cells to produce the recombinant virus, and healthy insect Sf9 cells were infected with the recombinant virus containing HBV polymerse gene to express the target protein. HBV polymerse expressed in insect cells was analyzed by SDS-PAGE. PCR results showed recombinant donor plasmid, pFastbac Dual-pol, was constructed successfully. The recombinant hepatitis B virus polymerase was expressed in insect cells at high level. The recombinant hepatitis B virus polymerase should facilitate the analysis of HBV polymerase biological characteristics, allow the investigation for new anti-HBV drugs specifically blocking HBV polymerase.

  13. Immunogenicity of two different dosages (10 and 5 μg) of recombinant DNA hepatitis B vaccine in healthy neonates

    NARCIS (Netherlands)

    R. Del Cancho (R.); P.M. Grosheie (P.); M. Voogd-Schotanus (M.); W. Huisman (Willem); R.A. Heijtink; S.W. Schalm (Solko)

    1994-01-01

    textabstractThe immunogenicity of a half (5 μg) and a full (10 μg) dosage of recombinant DNA yeast-derived hepatitis B vaccine (HB-Vax-DNA) in healthy neonates was assessed in order to compare two candidate dosages of vaccine. After randomization 174 newborns of HBsAg-negative mothers entered the st

  14. Evidence of intratypic recombination in natural populations of hepatitis C virus

    International Nuclear Information System (INIS)

    Hepatitis C virus (HCV) has high genomic variability and, since its discovery, at least six different types and an increasing number of subtypes have been reported. Genotype 1 is the most prevalent genotype found in South America. In the present study, three different genomic regions (5 UTR, core and NS5B) of four HCV strains isolated from Peruvian patients were sequenced in order to investigate the congruence of HCV genotyping for these three genomic regions. Phylogenetic analysis using 5 UTR-core sequences found strain PE22 to be related to subtype 1a. To test the possibility of genetic recombination, phylogenetic studies were carried out, revealing that a crossover event had taken place in the NS5B protein. We discuss the consequences of this observation on HCV genotype classification, laboratory diagnosis and treatment of HCV infection

  15. IMMUNOGENICITY OF LOW-DOSE INTRAMUSCULAR AND INTRADERMAL VACCINATION WITH RECOMBINANT HEPATITIS B VACCINE

    Directory of Open Access Journals (Sweden)

    TURCHI Marília Dalva

    1997-01-01

    Full Text Available The study is a randomized trial using recombinant DNA vaccine to determine whether an intramuscular 10 µg dose or intradermal 2 µg induces satisfactory anti-HBs levels compared to the standard dose of intramuscular 20 µg. participants were 359 healthy medical and nurse students randomly allocated to one of the three groups: Group I - IM 20 µg; Group II - IM 10 µg; Group III - ID 2 µg at 0, 1 and 6 months. Anti-HBs titres were measured after complete vaccine schedule by ELISA/Pasteur. Baseline variables were similar among groups and side effects were mild after any dose. Vaccinees in the IM-10 µg group had seroconversion rate and geometric mean titre (GMT 2344 IU L-1, not significant different from the IM-20 µg group (GMT 4570 IU L-1. On the contrary, 21.4% of the ID - 2 µg recipients mount antibody concentration below 10 IU L1 and GMT of 91 IU L-1, a statiscally significant difference compared with the standard schedule IM-20 µg (p < 0.001. A three dose regimen of half dosse IM could be considered an appropriate schedule to prevent hepatitis B in young health adults which is of relevance to the expansion of hepatitis B vaccine programme

  16. Inflammatory response of endothelial cells to hepatitis C virus recombinant envelope glycoprotein 2 protein exposure

    Directory of Open Access Journals (Sweden)

    Ana Carolina Urbaczek

    2014-09-01

    Full Text Available The hepatitis C virus (HCV encodes approximately 10 different structural and non-structural proteins, including the envelope glycoprotein 2 (E2. HCV proteins, especially the envelope proteins, bind to cell receptors and can damage tissues. Endothelial inflammation is the most important determinant of fibrosis progression and, consequently, cirrhosis. The aim of this study was to evaluate and compare the inflammatory response of endothelial cells to two recombinant forms of the HCV E2 protein produced in different expression systems (Escherichia coli and Pichia pastoris. We observed the induction of cell death and the production of nitric oxide, hydrogen peroxide, interleukin-8 and vascular endothelial growth factor A in human umbilical vein endothelial cells (HUVECs stimulated by the two recombinant E2 proteins. The E2-induced apoptosis of HUVECs was confirmed using the molecular marker PARP. The apoptosis rescue observed when the antioxidant N-acetylcysteine was used suggests that reactive oxygen species are involved in E2-induced apoptosis. We propose that these proteins are involved in the chronic inflammation caused by HCV.

  17. Effects of augmentation of liver regeneration recombinant plasmid on rat hepatic fibrosis

    Institute of Scientific and Technical Information of China (English)

    Qing Li; Dian-Wu Liu; Li-Mei Zhang; Bing Zhu; Yu-Tong He; Yong-Hong Xiao

    2005-01-01

    AIM: To investigate the effects of eukaryotic expression of plasmid on augmentation of liver regeneration (ALR) in rat hepatic fibrosis and to explore their mechanisms. METHODS: Ten rats were randomly selected from 50Wistar rats as normal control group. The rest were administered intraperitoneally with porcine serum twice weekly. After 8 wk, they were randomly divided into:model control group, colchicine group (Col), first ALR group (ALR1), second ALR group (ALR2). Then colchicine ALR recombinant plasmid were used to treat them respectively. At the end of the 4th wk, rats were killed.Serum indicators were detected and histopathological changes were graded. Expression of type Ⅰ, Ⅲ, collagen and TIMP-1 were detected by immunohisto-chemistry and expression of TIMP-1 mRNA was detected by semiquantified RT-PCR.RESULTS: The histologic examination showed that the degree of the rat hepatic fibrosis in two ALR groups was lower than those in model control group. Compared with model group, ALR significantly reduced the serum levels of ALT,AST, HA, LN, PCⅢ and Ⅳ (P<0.05). Immunohistochemical staining showed that expression of type Ⅰ, Ⅲ, collagen and TIMP-1 in two ALR groups was ameliorated dramatically compared with model group (Ⅰ collagen: 6.94±1.42, 5.80±1.66and 10.83±3.58 in ALR1, ALR2 and model groups, respectively;Ⅲ collagen: 7.18±1.95, 4.50±1.67 and 10.25±2.61,respectively; TIMP-1: 0.39±0.05, 0.20±0.06 and 0.53±0.12,respectively, P<0.05 or P<0.01). The expression level of TIMP-1 mRNA in the liver tissues was markedly decreased in two ALR groups compared with model group (TIMP-1mRNA/β-actin: 0.89±0.08, 0.65±0.11 and 1.36±0.11 in ALR1, ALR2 and model groups respectively, P<0.01).CONCLUSION: ALR recombinant plasmid has beneficial effects on rat hepatic fibrosis by enhancing regeneration of injured liver cells and inhibiting TIMP-1 expressions.

  18. Immunogenicity of two different dosages (10 and 5 μg) of recombinant DNA hepatitis B vaccine in healthy neonates

    OpenAIRE

    Del Cancho, R.; Grosheie, P.; Voogd-Schotanus, M.; Huisman, Willem; Heijtink, R.A.; Schalm, Solko

    1994-01-01

    textabstractThe immunogenicity of a half (5 μg) and a full (10 μg) dosage of recombinant DNA yeast-derived hepatitis B vaccine (HB-Vax-DNA) in healthy neonates was assessed in order to compare two candidate dosages of vaccine. After randomization 174 newborns of HBsAg-negative mothers entered the study. Neonates received four doses of either 10 or 5 μg hepatitis B vaccine, according to the DTP polio immunization schedule at months 3, 4, 5 and 11. No serious adverse reactions were observed; 15...

  19. Characterization of Hepatitis C Virus Recombinants with Chimeric E1/E2 Envelope Proteins and Identification of Single Amino Acids in the E2 Stem Region Important for Entry

    OpenAIRE

    Carlsen, Thomas H. R.; Scheel, Troels K. H.; Ramirez, Santseharay; Foung, Steven K. H.; Bukh, Jens

    2013-01-01

    The hepatitis C virus (HCV) envelope proteins E1 and E2 play a key role in host cell entry and represent important targets for vaccine and drug development. Here, we characterized HCV recombinants with chimeric E1/E2 complexes in vitro. Using genotype 1a/2a JFH1-based recombinants expressing 1a core-NS2, we exchanged E2 with functional isolate sequences of genotypes 1a (alternative isolate), 1b, and 2a. While the 1a-E2 exchange did not impact virus viability, the 2a-E2 recombinant was nonviab...

  20. Significance of Response to Hepatitis B Recombinant Vaccine in Subjects with Isolated Antibody to Hepatitis B Core Antigen

    OpenAIRE

    Bahari, Ali; Izadi, Shahrokh; Bari, Zohreh; Khosravi, Soheyla; Baghaei, Bita; Saneimoghadam, Esmaeil; Firouzi, Farzad; Espiari, Ali; Esmaeilzadeh, Abbas; Mokhtarifar, Ali; Bakhshipour, Alireza; Ganji, Azita

    2015-01-01

    BACKGROUND It is important to differentiate whether isolated anti-HBc is due to false positive results or the prior exposure to hepatitis B virus, because individuals with false-positive anti-HBc can benefit from vaccination and their blood can be safely transfused. To distinguish between these two conditions, we evaluated the serologic response to hepatitis B vaccine. METHODS Ninety subjects with isolated anti-HBc (cases) and 100 subjects with totally negative hepatitis B serologic markers (...

  1. TH1 and TH2 responses are influenced by HLA antigens in healthy neonates vaccinated with recombinant hepatitis B vaccine.

    OpenAIRE

    Abdollah Jafarzadeh; Fazel Shokri

    2012-01-01

    The immune response to hepatitis B surface antigen (HBsAg) is influenced by several factors, of which HLA antigens and balanced secretion of Th1/Th2 cytokines play important roles. The aim of this study was to evaluate the influence of HLA antigens on cytokine secretion by HBsAg-stimulated peripheral blood mononuclear cells (PBMC) from healthy neonates vaccinated with recombinant HBsAg. PBMCs were isolated from 48 Iranian neonates vaccinated with a recombinant HBV vaccine. The cells were stim...

  2. Comparison of immunogenicity of Aluminum salts as adjuvant for recombinant Hepatitis-B vaccine

    Directory of Open Access Journals (Sweden)

    Fazeli MR

    2007-05-01

    Full Text Available Background: Aluminum salts are common adjuvants in human and animal vaccine preparations. The two adjuvants aluminum phosphate and aluminum hydroxide show acceptable immunoadjuvant properties with many antigens. These two salts have different physicochemical characteristics that make each one suitable for certain antigens. The surface antigen of Hepatitis B (HBsAg has several antigenic epitopes that bind to aluminum adjuvants by a ligand exchange mechanism. Although HBV vaccines using an aluminum hydroxide adjuvant are available, higher antigenicity is needed for the subgroup of people who do not respond sufficiently to the currently available vaccines. Methods: A solution of recombinant HBsAg for making different formulations of vaccines with aluminum phosphate (Adju-Phos® and aluminum hydroxide (Alhydrogel® adjuvants was obtained from Darupakhsh Pharmaceutical Company. The total protein content, antigenicity, and purity of HBsAg solution were determined using BCA, ELISA, and SDS-PAGE methods, respectively. The different formulations were prepared in the lab and administered i.p. to two test groups of Balb/C mice and a third test group received the Engerix vaccine, which is currently available on the market and uses an aluminum hydroxide adjuvant. The control group of animals received the solution without antigen. After 28 days, heart blood samples were collected and serum was separated to determine the antibody titer against HBsAg using an ELISA kit. Results: This study shows that the vaccine formulated with aluminum phosphate exerted more immunogenicity than both the aluminum hydroxide laboratory formulation and the Engerix vaccines. Conclusion: Although the results of our study indicate higher immunogenic properties of the vaccine formulated with the aluminum phosphate adjuvant, complementary experiments are needed to further evaluate the biological properties with respect to effectiveness, adverse effects, product stability and finally

  3. Multicenter study on the immunogenicity and safety of two recombinant vaccines against hepatitis B

    Directory of Open Access Journals (Sweden)

    Reinaldo Menezes Martins

    2004-12-01

    Full Text Available The immunogenicity and safety of a new recombinant hepatitis B vaccine from the Instituto Butantan (Butang® were evaluated in a multicenter, double-blind, prospective equivalence study in three centers in Brazil. Engerix B® was the standard vaccine. A total of 3937 subjects were recruited and 2754 (70% met all protocol criteria at the end of the study. All the subjects were considered healthy and denied having received hepatitis B vaccine before the study. Study subjects who adhered to the protocol were newborn infants (566, children 1 to 10 years old (484, adolescents from 11 to 19 years (740, adults from 20 to 30 years (568, and adults from 31 to 40 years (396. Vaccine was administered in three doses on the schedule 0, 1, and 6 months (newborn infants, adolescents, and adults or 0, 1, and 7 months (children. Vaccine dose was intramuscular 10 µg (infants, children, and adolescents or 20 µg (adults. Percent seroprotection (assumed when anti-HBs titers were > 10mIU/ml and geometric mean titer (mIU/ml were: newborn infants, 93.7% and 351.1 (Butang® and 97.5% and 1530.6 (Engerix B®; children, 100% and 3600.0 (Butang® and 97.7% and 2753.1 (Engerix B®; adolescents, 95.1% and 746.3 (Butang® and 96% and 1284.3 (Engerix B®; adults 20-30 years old, 91.8% and 453.5 (Butang® and 95.5% and 1369.0 (Engerix B®; and adults 31-40 years old, 79.8% and 122.7 (Butang® and 92.4% and 686.2 (Engerix B®. There were no severe adverse events following either vaccine. The study concluded that Butang® was equivalent to Engerix B® in children, and less immunogenic but acceptable for use in newborn infants, adolescents, and young adults.

  4. Recombinant Hepatitis B Vaccine Adjuvanted With AS04 in Dialysis Patients: A Prospective Cohort Study

    Directory of Open Access Journals (Sweden)

    Fabrizio Fabrizi

    2015-11-01

    Full Text Available Background/Aims: Patients undergoing maintenance dialysis have an unsatisfactory response to vaccination, including to hepatitis B vaccine. A recombinant HB vaccine containing a new adjuvant system AS04 (HBV-AS04 has been recently developed; a few data exist on the immunogenicity and safety of HBV-AS04 among patients undergoing regular dialysis. All hepatitis B virus-seronegative patients with undetectable antibody against HBsAg undergoing maintenance dialysis at two units were prospectively included. Methods: Patients received four 20-mcg doses of HBV-AS04 by intramuscular route (deltoid muscle at months 0,1,2, and 3. Anti-HB surface antibody concentrations were measured at intervals of 1, 2, 3, 4, and 12 months. Univariate and multivariate analyses determined which parameters predicted immunologic response to HBV-AS04 vaccine. Results: 102 patients were enrolled and 91 completed the study. At completion of the vaccination schedule, using per-protocol analysis, 76 of 91 (84% had antibody titers ≥10 mIU/mL with anti-HBs geometric antibody concentrations (GMCs of 385.25 mIU/mL. The sero-protection rate at month 12 was 84% (48/57 with lower GMCs (62.74 mIU/mL, PPConclusions: HBV-AS04 vaccine was highly immunogenic in our cohort of patients on maintenance dialysis even if a significant number of non-responders is still present. Prospective studies with HBV-AS04 on larger study groups and with longer follow-ups are under way.

  5. Inhibition of hepatitis B virus DNA replicative intermediate forms by recombinant interferon-γ

    Institute of Scientific and Technical Information of China (English)

    Mohammad Khalid Parvez; Deepak Sehgal; Shiv Kumar Sarin; Seemi Farhat Basir; Shahid Jameel

    2006-01-01

    AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-γ, alone and in combination with lamivudine.METHODS: A recombinant baculovirus-HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization.RESULTS: IFN-γ at 0.1 to 5 μg/L efficiently down regulated HBsAg expression in transduced HepG2 cells.At 5 μg/L, IFN-γ also suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-γ showed no additive effect,sequential treatment first with lamivudine and then IFN-γ was found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2 μmol/L lamivudine for two days, followed by 1 μg/L IFN-γ for another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2 μmol/L lamivudine for two days, followed by 5 μg/L IFN-γ for six days showed a 72% reduction in HBV cccDNA pool.CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-γ and lamivudine,especially in IFN-α non-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants.

  6. Evidence for Protection against Chronic Hepatitis C Virus Infection in Chimpanzees by Immunization with Replicating Recombinant Vaccinia Virus▿

    OpenAIRE

    Youn, Jin-Won; Hu, Yu-Wen; Tricoche, Nancy; Pfahler, Wolfram; Shata, Mohamed Tarek; Dreux, Marlene; Cosset, François-Loic; Folgori, Antonella; Lee, Dong-Hun; Brotman, Betsy; Prince, Alfred M.

    2008-01-01

    Given the failures of nonreplicating vaccines against chronic hepatitis C virus (HCV) infection, we hypothesized that a replicating viral vector may provide protective immunity. Four chimpanzees were immunized transdermally twice with recombinant vaccinia viruses (rVV) expressing HCV genes. After challenge with 24 50% chimpanzee infective doses of homologous HCV, the two control animals that had received only the parental VV developed chronic HCV infection. All four immunized animals resolved...

  7. PRELIMINARY REPORT OF THE USE ON ADULTS OF A RECOMBINANT YEAST-DERIVED HEPATITIS B VACCINE MANUFACTURED BY INSTITUTO BUTANTAN

    OpenAIRE

    Angela Aparecida COSTA; Marta INENAMI; Edmundo JUAREZ; LLACEN Pedro Dorlhiac; Raw, Isaias

    1997-01-01

    Three 10 µg doses of the recombinant hepatitis B vaccine, manufactured by Instituto Butantan by original technology, were administered in a adult population, mean age 30 years old, following the 0, 1 and 6 months schedule immunization. The clinical trial was considered satisfactory in terms of immunogenicity (anti-HBs titers between 17.5-29500 IU/l, seroconversion 95.3%) and reactogenicity (no incapacitating side effects)Três doses de 10 µg da vacina recombinante contra hepatite B, produzida ...

  8. Immunogenicity and reactogenicity of a recombinant hepatitis B vaccine in subjects over age of forty years and response of a booster dose among nonresponders

    OpenAIRE

    Das, Kunal; Gupta, R.K.; Kumar, V.; Kar, P.

    2003-01-01

    AIM: The study was initiated to evaluate the reactogenicity and immunogenicity of a recombinant hepatitis B vaccine in age group > 40 years and to study the response of a single booster dose in primary non-responders to the hepatitis B vaccination.

  9. HCV prototype vaccine based on hepatitis B core virus-like particles

    OpenAIRE

    Marija Mihailova

    2008-01-01

    HCV prototype vaccine based on hepatitis B core virus-like particles Abstract In the current study the C-terminally truncated HBc expression vectors were used for exposure of different hepatitis C virus (HCV) protein (core, E2, and NS3) fragments. All created chimeric constructs directed high level of recombinant protein synthesis in E.coli. However, not all chimeric proteins were able to self-assemble into virus-like particles (VLPs). HBcCterm/HVR1tetramer VLPs turned ou...

  10. Simple high-cell density fed-batch technique for high-level recombinant protein production with Pichia pastoris: Application to intracellular production of Hepatitis B surface antigen

    Directory of Open Access Journals (Sweden)

    Ross Anton

    2009-02-01

    Full Text Available Abstract Background Hepatitis B is a serious global public health concern. Though a safe and efficacious recombinant vaccine is available, its use in several resource-poor countries is limited by cost. We have investigated the production of Hepatitis B virus surface antigen (HBsAg using the yeast Pichia pastoris GS115 by inserting the HBsAg gene into the alcohol oxidase 1 locus. Results Large-scale production was optimized by developing a simple fed-batch process leading to enhanced product titers. Cells were first grown rapidly to high-cell density in a batch process using a simple defined medium with low salt and high glycerol concentrations. Induction of recombinant product synthesis was carried out using rather drastic conditions, namely through the addition of methanol to a final concentration of 6 g L-1. This methanol concentration was kept constant for the remainder of the cultivation through continuous methanol feeding based on the on-line signal of a flame ionization detector employed as methanol analyzer in the off-gas stream. Using this robust feeding protocol, maximum concentrations of ~7 grams HBsAg per liter culture broth were obtained. The amount of soluble HBsAg, competent for assembly into characteristic virus-like particles (VLPs, an attribute critical to its immunogenicity and efficacy as a hepatitis B vaccine, reached 2.3 grams per liter of culture broth. Conclusion In comparison to the highest yields reported so far, our simple cultivation process resulted in an ~7 fold enhancement in total HBsAg production with more than 30% of soluble protein competent for assembly into VLPs. This work opens up the possibility of significantly reducing the cost of vaccine production with implications for expanding hepatitis B vaccination in resource-poor countries.

  11. Efficacy of Double Dose Recombinant Hepatitis B Vaccination in Chronic Hepatitis C Patients, Compared to Standard Dose Vaccination

    OpenAIRE

    Mohammad Minakari; Afshin Tahmasebi; Mahyar Hosseini Motlagh; Behrooz Ataei; Majid Yaran; Hamid Kalantari; Hamid Tavakkoli

    2014-01-01

    Background: Hepatitis B virus (HBV) vaccination is a well-known, safe and effective way for protection against HBV infection; however, non-responders remain susceptible to infection with HBV. This is so important in patients with any kind of chronic liver disease, especially chronic hepatitis C virus (HCV) patients in whom acute HBV infection may lead to decompensation of liver disease. Some of the studies have shown that immunogenicity of HBV vaccination is decreased in these patients. The a...

  12. Purification of hepatitis B surface antigen virus-like particles from recombinant Pichia pastoris and in vivo analysis of their immunogenic properties.

    OpenAIRE

    Gurramkonda, Chandrasekhar; Zahid, Maria; Nemani, Satish Kumar; Adnan, Ahmad; Gudi, Satheesh Kumar; Khanna, Navin; Ebensen, Thomas; Lünsdorf, Heinrich; Guzmán, Carlos A.; Rinas, Ursula

    2013-01-01

    Following earlier studies on high-level intracellular production of hepatitis B surface antigen (HBsAg) using recombinant Pichia pastoris, we present here in detail an enhanced method for the purification of recombinant HBsAg virus-like particles (VLPs). We have screened various detergents for their ability to promote the solubilization of recombinant intracellular HBsAg. In addition, we have analyzed the effect of cell disruption and extraction regarding their impact on the release of HBsAg....

  13. Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors

    Directory of Open Access Journals (Sweden)

    Ana Carolina Urbaczek

    2015-06-01

    Full Text Available Hepatitis C virus (HCV envelope protein 2 (E2 is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr and CD81 in human umbilical vein endothelial cells (HUVEC and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients.

  14. Productive homologous and non-homologous recombination of hepatitis C virus in cell culture

    DEFF Research Database (Denmark)

    Scheel, Troels K H; Galli, Andrea; Li, Yi-Ping;

    2013-01-01

    . In addition, recombination is an important regulatory mechanism of cytopathogenicity for the related pestiviruses. Here we describe recombination of HCV RNA in cell culture leading to production of infectious virus. Initially, hepatoma cells were co-transfected with a replicating JFH1ΔE1E2 genome (genotype 2a...... incompetent genomes were co-transfected. Reverse genetic studies verified the viability of representative recombinants. After serial passage, subsequent recombination events reducing or eliminating the duplicated region were observed for some but not all recombinants. Furthermore, we found that inter......-genotypic recombination could occur, but at a lower frequency than intra-genotypic recombination. Productive recombination of attenuated HCV genomes depended on expression of all HCV proteins and tolerated duplicated sequence. In general, no strong site specificity was observed. Non-homologous recombination was observed...

  15. Characterization of hepatitis C virus envelope glycoprotein complexes expressed by recombinant vaccinia viruses.

    Science.gov (United States)

    Ralston, R; Thudium, K; Berger, K; Kuo, C; Gervase, B; Hall, J; Selby, M; Kuo, G; Houghton, M; Choo, Q L

    1993-11-01

    We constructed recombinant vaccinia virus vectors for expression of the structural region of hepatitis C virus (HCV). Infection of mammalian cells with a vector (vv/HCV1-906) encoding C-E1-E2-NS2 generated major protein species of 22 kDa (C), 33 to 35 kDa (E1), and 70 to 72 kDa (E2), as observed previously with other mammalian expression systems. The bulk of the E1 and E2 expressed by vv/HCV1-906 was found integrated into endoplasmic reticulum membranes as core-glycosylated species, suggesting that these E1 and E2 species represent intracellular forms of the HCV envelope proteins. HCV E1 and E2 formed E1-E2 complexes which were precipitated by either anti-E1 or anti-E2 serum and which sedimented at approximately 15 S on glycerol density gradients. No evidence of intermolecular disulfide bonding between E1 and E2 was detected. E1 and E2 were copurified to approximately 90% purity by mild detergent extraction followed by chromatography on Galanthus nivalus lectin-agarose and DEAE-Fractogel. Immunization of chimpanzees with purified E1-E2 generated high titers of anti-E1 and anti-E2 antibodies. Further studies, to be reported separately, demonstrated that purified E1-E2 complexes were recognized at high frequency by HCV+ human sera (D. Y. Chien, Q.-L. Choo, R. Ralston, R. Spaete, M. Tong, M. Houghton, and G. Kuo, Lancet, in press) and generated protective immunity in chimpanzees (Q.-L. Choo, G. Kuo, R. Ralston, A. Weiner, D. Chien, G. Van Nest, J. Han, K. Berger, K. Thudium, J. Kansopon, J. McFarland, A. Tabrizi, K. Ching, B. Mass, L. B. Cummins, E. Muchmore, and M. Houghton, submitted for publication), suggesting that these purified HCV envelope proteins display native HCV epitopes. PMID:8411378

  16. Near-Full Genome Characterisation of Two Natural Intergenotypic 2k/1b Recombinant Hepatitis C Virus Isolates

    Directory of Open Access Journals (Sweden)

    Victoria L. Demetriou

    2011-01-01

    Full Text Available Few natural intergenotypic hepatitis C virus (HCV recombinants have been characterised, and only RF1_2k/1b has demonstrated widespread transmission. The near-full length genome sequences for two cases of 2k/1b recombinants (CYHCV037 and CYHCV093 sampled in Cyprus were obtained using strain-specific RT-PCR amplification and sequencing protocols. Sequence analysis confirmed their similarity with the original RF1_2k/1b strain from St. Petersburg, N687. These two isolates significantly contribute to the sequence data available on this recombinant and confirm its increasing spread among individuals from Eastern Europe, and its association with transmission through intravenous drug use. Phylogenetic analyses reveal clustering of the sequence 3′ to the recombination point, not seen in the topology of the 5′ sequences, implying a more complicated evolutionary history than that held to date. The increasing cases of HCV recombinant strains underline the requirement of their contribution to the standardised rules of HCV classification and nomenclature, molecular epidemiology, diagnosis, and treatment.

  17. Central nervous system demyelinating diseases and recombinant hepatitis B vaccination: a critical systematic review of scientific production.

    Science.gov (United States)

    Martínez-Sernández, V; Figueiras, A

    2013-08-01

    The etiology of multiple sclerosis has not yet been fully described. A potential link between the recombinant hepatitis B vaccine and an increased risk of onset or exacerbation of multiple sclerosis emerged in the mid-1990s, leading to several spontaneous reports and studies investigating this association. We conducted a critical systematic review aimed at assessing whether hepatitis B vaccination increases the risk of onset or relapse of multiple sclerosis and other central nervous system demyelinating diseases. MEDLINE and EMBASE were used as data sources, and the search covered the period between 1981 and 2011. Twelve references met the inclusion criteria. No significant increased risk of onset or relapse of the diseases considered was associated with hepatitis B vaccination, except in one study. Most studies included in this review displayed methodological limitations and heterogeneity among them, which rendered it impossible to draw robust conclusions about the safety of hepatitis B vaccination in healthy subjects and patients with multiple sclerosis. Therefore, on the basis of current data there is no need to modify the vaccination recommendations; however, there is a need to improve the quality of observational studies with emphasis on certain considerations that are discussed in this review. PMID:23086181

  18. Epidermal growth factor receptor antibody plus recombinant human endostatin in treatment of hepatic metastases after remnant gastric cancer resection

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    We report a 55-year-old male who developed advanced hepatic metastasis and peritoneal carcinomatosis after resection of remnant gastric cancer resection 3 mo ago. The patient only received epidermal growth factor (EGF) receptor antibody (Cetuximab) plus recombinant human endostatin (Endostar).Anti-tumor activity was assessed by 18F-fluorodeoxyglucose (18F-FDG)positron emission tomography/computer tomography (PET/CT) at baseline and then every 4 wk. The case illustrates that 18FDG-PET/CT could make an early prediction of the response to Cetuximab plus Endostar in such clinical situations. 18FDG-PET/CT is a useful molecular imaging modality to evaluate the biological response advanced hepatic metastasis and peritoneal carcinomatosis to Cetuximab plus Endostar in patients after remnant gastric cancer resection.

  19. Significance of Response to Hepatitis B Recombinant Vaccine in Subjects with Isolated Antibody to Hepatitis B Core Antigen.

    Science.gov (United States)

    Bahari, Ali; Izadi, Shahrokh; Bari, Zohreh; Khosravi, Soheyla; Baghaei, Bita; Saneimoghadam, Esmaeil; Firouzi, Farzad; Espiari, Ali; Esmaeilzadeh, Abbas; Mokhtarifar, Ali; Bakhshipour, Alireza; Ganji, Azita

    2015-10-01

    BACKGROUND It is important to differentiate whether isolated anti-HBc is due to false positive results or the prior exposure to hepatitis B virus, because individuals with false-positive anti-HBc can benefit from vaccination and their blood can be safely transfused. To distinguish between these two conditions, we evaluated the serologic response to hepatitis B vaccine. METHODS Ninety subjects with isolated anti-HBc (cases) and 100 subjects with totally negative hepatitis B serologic markers (controls) were recruited to receive three doses of hepatitis-B (HB) vaccine. Thirty days after the first dose of the vaccine, anti-HBs titers were checked and individuals with anti-HBs titer >50 mIU/mL did not receive additional doses of the vaccine. However, others completed the vaccination course, and another blood sample was collected 30 days after the third dose to measure anti-HBs level. RESULTS Nineteen (21.1%) cases and three (3%) controls had no sero-conversion (anti-HBs titers vaccine, but the rate was significantly lower (5%) in the control group (pvaccination. CONCLUSION More than 75% of individuals with positive isolated anti-HBc can benefit from vaccination and can be included in donor pool. Also, one fifth seemed to have occult HBV infection. So HB vaccination may be used as a diagnostic tool for clarifying the situation of the subjects with isolated anti-HBc. PMID:26609352

  20. Small-angle neutron scattering study of recombinant yeast-derived human hepatitis B virus surface antigen vaccine particle

    Science.gov (United States)

    Sato, M.; Ito, Y.; Kameyama, K.; Imai, M.; Ishikawa, N.; Takagi, T.

    1995-02-01

    The overall and internal structure of recombinant yeast-derived human hepatitis B virus surface antigen vaccine particles was investigated by small-angle neutron scattering using the contrast variation method. The vaccine is a nearly spherical particle, and its contrast-matching point was determined to be at about 24% D 2O content, indicating that a large part of the vaccine particle is occupied by lipids and carbohydrates from the yeast. The Stuhrmann plot suggests that the surface antigens exist predominantly in the peripheral region of the particle, which is favorable to the induction of anti-virus antibodies.

  1. A recombinant antibody-interleukin 2 fusion protein suppresses growth of hepatic human neuroblastoma metastases in severe combined immunodeficiency mice.

    OpenAIRE

    Sabzevari, H; Gillies, S D; Mueller, B M; Pancook, J D; Reisfeld, R A

    1994-01-01

    A genetically engineered fusion protein consisting of a human/mouse chimeric anti-ganglioside GD2 antibody (ch14.18) and recombinant human interleukin 2 (rhIL-2) was tested for its ability to target rhIL-2 to tumor sites and stimulate immune effector cells sufficiently to achieve effective tumor cell lysis in vivo. The ch14.18-IL-2 fusion protein proved more effective than equivalent doses of rhIL-2 in suppressing dissemination and growth of human neuroblastoma in an experimental hepatic meta...

  2. Long-term persistence of immunity after vaccination of pre-adolescents with low doses of a recombinant hepatitis B vaccine

    OpenAIRE

    Gilca, Vladimir; De Serres, Gaston; Boulianne, Nicole; Murphy, Donald; Ouakki, Manale; De Wals, Phillipe; Trudeau, Gisele; Massé, Richard; Dionne, Marc

    2013-01-01

    Background and aims: Recent studies have shown no detectable antibodies and no response to a challenge dose of vaccine 10–20 y after receiving low doses (2.5–5 µg) of recombinant hepatitis B vaccine during first months of life. Little information is available on long-term persistence of immunity after vaccinating pre-adolescents with low doses of hepatitis B vaccine.

  3. The effect of zinc sulfate on immunologic response to recombinant hepatitis B vaccine in elderly

    OpenAIRE

    Afsharian, Mandana; Vaziri, Siavash; Janbakhsh, Ali Reza; Sayad, Babak; Mansouri, Feizollah; Nourbakhsh, Javad; Qadiri, Keyghobad; Najafi, Farid; Shirvanii, Maria

    2011-01-01

    Background Hepatitis B is the major cause of chronic hepatitis and cirrhosis in Iran. Sanitation and immunization is one of the most effective measures for prevention of the disease which is now widely used in developing countries. However, the immune response to the vaccine varies by age. Objectives To determine the effect of zinc sulfate on immune response to hepatitis-B vaccine in elderly. Patients and Methods In a clinical trial on 140 subjects aged ?40 years with a body mass index (BMI)

  4. Recombinant Human Hepatitis B Vaccine Initiating Alopecia Areata: Testing the Hypothesis Using the C3H/HeJ Mouse Model

    OpenAIRE

    Sundberg, John P; Silva, Kathleen A.; Zhang, Weidong; Sundberg, Beth A.; Edwards, Kathryn; King, Lloyd E.; Davis, Robert L; Black, Steven

    2009-01-01

    Untoward effects of human vaccines suggest that recombinant hepatitis B vaccine may induce alopecia areata (AA) in some patients. Similar untoward immunological effects may also account for AA-like diseases in domestic species. In this study the C3H/HeJ spontaneous adult onset AA mouse model was used to test the role, if any, of recombinant hepatitis B vaccine on the initiation or activation of AA. Initial experiments demonstrated no effect on induction of AA in young adult female C3H/HeJ mic...

  5. [Serologic response to a DNA recombinant vaccine against hepatitis B in natives of the Peruvian Amazonian jungle].

    Science.gov (United States)

    Colichón, A; Vildósola, H; Sjogren, M; Cantella, R; Rojas, C

    1990-01-01

    Large areas of the Amazon basin in Brazil, Colombia, Ecuador, and in the nonoriental region of the peruvian jungle have been found to be hyperendemic to Hepatitis B with high prevalence of asymptomatic carriers (11 to 25%) and, in more selected areas, Hepatitis Delta has been also reported. In the present report, we have studied 108 volunteers from six different Jivaroes communities living in a hyperendemic Hepatitis B area. They received 2 doses of DNA recombinant yeast derivated HBV vaccine. All the selected persons were HBsAb negatives, but many (80%) had antibodies to HBc. Following immunization schedule, 80% responded with the formation of HBsAb; a better seroconversion was achieved in those negatives to anticore IgG compared with those having HBcAb. We obtained 90% of seroconversion in spite of the fact that our vaccination schedule was prolonged up to 10 months from the one recommended by the manufacturer. The vaccination schedule 0,4, 14 months, and the schedule 0,4 months, had 76 and 29% of seroconversion, respectively. We want to point out three observations: 1) It is quite possible that many of the Anti-core positives, that did not respond to vaccination were carriers of HBsAg undetectable by the conventional EIA test carried out; 2) The seroconversion rate in these natives was low (up to six months after the vaccination schedule); and 3) Many of the HBcAb were false positives and many of them were recently infected. We conclude: A) It is highly important to assess the anti-HBs hyperendemic areas before attempting vaccinations; B) All persons negative to anti-HBs should be vaccinated in spite to anticore antibodies; C) Areas with difficult access could be vaccinated even until 10 months without affecting good results, and D) DNA recombinant vaccine (ENGERIX B) was well tolerated. No side effects were observed.

  6. A candidate therapeutic vaccine against hepatitis C virus infection: Use of a recombinant alphavirus vector

    OpenAIRE

    Ip, Peng

    2014-01-01

    Peng Peng Ip beschrijft in dit proefschrift haar onderzoek naar de ontwikkeling van een immuuntherapie gericht tegen hepatitis C virus infecties. Wereldwijd zijn ongeveer 150 miljoen mensen chronisch besmet met hepatitis C virus (HCV) en jaarlijks sterven 350.000 mensen aan een HCV-gerelateerde leverziekte zoals cirrose of leverkanker. In Nederland zijn ongeveer 60.000 mensen besmet met HCV, vaak nog zonder het te weten. Het doel van het promotie onderzoek was om een vaccin tegen HCV infectie...

  7. Transient facial nerve paralysis (Bell's palsy) following administration of hepatitis B recombinant vaccine: a case report.

    Science.gov (United States)

    Paul, R; Stassen, L F A

    2014-01-01

    Bell's palsy is the sudden onset of unilateral transient paralysis of facial muscles resulting from dysfunction of the seventh cranial nerve. Presented here is a 26-year-old female patient with right lower motor neurone facial palsy following hepatitis B vaccination. Readers' attention is drawn to an uncommon cause of Bell's palsy, as a possible rare complication of hepatitis B vaccination, and steps taken to manage such a presentation.

  8. Set up of analytical methods for evaluation of specifications of recombinant Hepatitis-B vaccine

    OpenAIRE

    Daram M; Akbari M; Norouzi M; Ajdary S; Mahmoudi M; Pakzad SR; Karimzadeh H; Jazayeri SM

    2009-01-01

    "nBackground: Hepatitis B vaccination has been included in routine immunization of all individuals according to WHO recommendations since 1991. Despite successful coverage, 3-5% of recipients fail to mount a desirable protection level of Ab. Vaccine failure results from: emergence of mutation, immune failure of individuals, decrease in vaccine potency, and etc. The quality of Hepatitis B vaccine should be evaluated by a reliable method. "n"nMethods: The amount of vaccine antige...

  9. "PERSISTENCE OF ANTI-HBs ANTIBODIES IN HEALTHY IRANIAN CHILDREN VACCINATED WITH RECOMBINANT HEPATITIS B VACCINE AND RESPONSE TO A BOOSTER DOSE"

    OpenAIRE

    Jafarzadeh, A; S. M. A. Sajjadi

    2005-01-01

    Long-term protection against hepatitis B virus (HBV) is dependent on persistence of anti-HBs antibodies and/or strong immunological memory. In this study we evaluated the persistence of anti-HBs antibodies in healthy Iranian children 5 years after primary vaccination and the response to a booster dose using recombinant hepatitis B vaccine. Totally, 81 children who had received primary course of hepatitis B vaccine at 0, 1.5 and 9 months of age were included in this study. A booster dose of he...

  10. Psychiatric adverse effects induced by recombinant interferon alfa in patients with chronic hepatitis C

    Directory of Open Access Journals (Sweden)

    Nešić Zorica I.

    2004-01-01

    Full Text Available Introduction Hepatitis C virus infection is a slowly progressive chronic disease and the most common cause of chronic liver disease. Presently, interferon alfa based therapies, with or without ribavirin, are standard treatment for chronic hepatitis C virus infection. The most troublesome psychiatric side effects of interferon therapy in our patients are: insomnia, irritability, anxiety, mood changes, short temper, emotional and affective lability, impaired cognitive function, apathy, loss of motivation and the most serious depression with or without suicidal ideas. Material and methods In our study we treated 82 patients chronically infected with HCV divided into 3 groups: the first group of 31 patients (20 male and 11 female received IFN-alfa in standard doses of 3 MU three times a week (t.i.w during 24 weeks; the second group of 36 patients (25 male and 11 female received IFN-alfa, 3 MU t.i.w plus ribavirin 1000-1200 mg per day during 24 weeks; the third group of 15 patients (11 male and 4 female received IFN-alfa, 3 MU t.i.w plus ribavirin 1000-1200 mg per day during 48 weeks. The follow-up period after therapy in all groups lasted 24 weeks. Results During treatment, we observed at least one psychiatric side effect in 21 (26% patients: insomnia in 11 (13%, emotional and affective lability in 9 (11%, anxiety, irritability and short temper in 8 (10%, impaired cognitive function in 7 (8% apathia and loss of motivation in 6 (7% treated patients. Depression, the most serious side effect, was established in 8 (10% patients. All of these side effects were observed during later stages of treatment (between 5th and 22nd weeks of treatment. The incidence of all psychiatric side effects was significantly higher in women than in men (p < 0,01. We observed higher prevalence of depression among patients with history of alcohol and drug abuse. Treatment was temporarily discontinued (from 2 to 4 weeks in all patients with depression, but it was not

  11. Neutralization resistance of hepatitis C virus can be overcome by recombinant human monoclonal antibodies

    DEFF Research Database (Denmark)

    Pedersen, Jannie L; Carlsen, Thomas H R; Prentoe, Jannick;

    2013-01-01

    Immunotherapy and vaccine development for hepatitis C virus (HCV) will depend on broadly reactive neutralizing antibodies (NAbs). However, studies in infectious strain JFH1-based culture systems expressing patient-derived Core-NS2 proteins have suggested neutralization resistance for specific HCV......-derived genotype 2a (strain T9), 2b (strains DH8 and DH10), and 2c (strain S83) consensus sequences, were viable in Huh7.5 hepatoma cells without requirement for adaptive mutations, reaching HCV infectivity titers of 3.9-4.5 log10 focus-forming units per milliliter. In in vitro neutralization assays, we...... demonstrated that the novel genotype 2 viruses as well as prototype strains J6/JFH1(2a) and J8/JFH1(2b), all with authentic envelope proteins, were resistant to neutralization by genotype 2a, 2b, 2c, 2j, 2i, and 2q patient sera. However, these patient sera had high titers of HCV-specific NAbs, because...

  12. Differential sensitivity of 5'UTR-NS5A recombinants of hepatitis C virus genotypes 1-6 to protease and NS5A inhibitors

    DEFF Research Database (Denmark)

    Li, Yi-Ping; Ramirez, Santseharay; Humes, Daryl;

    2014-01-01

    BACKGROUND & AIMS: Hepatitis C virus (HCV) therapy will benefit from the preclinical evaluation of direct-acting antiviral (DAA) agents in infectious culture systems that test the effects on different virus genotypes. We developed HCV recombinants comprising the 5' untranslated region-NS5A (5-5A)...

  13. Set up of analytical methods for evaluation of specifications of recombinant Hepatitis-B vaccine

    Directory of Open Access Journals (Sweden)

    Daram M

    2009-06-01

    Full Text Available "nBackground: Hepatitis B vaccination has been included in routine immunization of all individuals according to WHO recommendations since 1991. Despite successful coverage, 3-5% of recipients fail to mount a desirable protection level of Ab. Vaccine failure results from: emergence of mutation, immune failure of individuals, decrease in vaccine potency, and etc. The quality of Hepatitis B vaccine should be evaluated by a reliable method. "n"nMethods: The amount of vaccine antigen was measured through the in vitro assay of Hepatitis B vaccines which consists of multiple dilutions of the reference material and samples. The preparations were evaluated by Elisa to determine the amount of HBsAg. The data were analyzed by parallel-line analysis software. The in vivo assay was performed by inoculating multiple doses of the reference and sample preparations in Balb/c mice. A control group was also inoculated with vaccine matrix. Four weeks later, the mice sera were evaluated to determine the presence of antibodies against Hepatitis B by Elisa method. The data were analyzed by Probit analysis software. "n"nResults: Both methods were set up in our laboratory by which different batches of Hepatitis B vaccine were evaluated. It was observed that In vivo and In vitro methods provide comparable results. Therefore we can use the in vitro method for routine testing of HB vaccine quality control. "n"nConclusion: In vitro method can be used in place of In vivo method because of its time and cost-effectiveness. Moreover, since no animals are used in in vitro method, it complies well with the 3R concept (Reduction, Refinement, and Replacement of animal testing and the current tendency to use alternative method.

  14. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    2010349 Relationships between serum hepatitis B virus load in mothers,free maternal DNA in peripheral blood of newborns and hepatitis B virus infection of newborns. WEI Junni(魏俊妮),et al. Dept Epidemiol,Shanxi Med Univ,Taiyuan 030001. Chin J Infect Dis 2010;28(5):297-300. Objective To study the relationships between serum hepatitis B virus (HBV) DNA level

  15. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    1993-01-01

    930140 Hepatocyte stimulator peptide and itsclinical significance in viral hepatitis.ZHOUWeiping(周卫平),et al.Instit Viral Hepatitis,Chongqing Med Univ,630010.Chin J InternMed 1992;31(10):626-628.Hepatocyte stimulator peptide(HSP)is anewly developed hepatic stimulator substance.Its monoclonal antibodies have been obtained inour laboratory.In this study,HSP was deter-mined in the sera of 315 subjects including pa-

  16. Immune responses in patients with HIV infection after vaccination with recombinant Hepatitis B virus vaccine

    Directory of Open Access Journals (Sweden)

    Singh Haqeeqat

    2006-03-01

    Full Text Available Abstract Background Patients with HIV infection are at risk of co-infection with HBV, as the routes of transmission are shared and thus immunization with HBV vaccine could be protective in them. The aim of the present study was to assess the efficacy of recombinant vaccine in treatment-naive HIV positive patients and healthy controls, and to dissect out differences if any, in different limbs of immune response. Methods Forty HIV positive patients and 20 HIV negative controls, negative for HBsAg, HBsAbs and HBcAbs were vaccinated with three doses of 40μg and 20μg of vaccine respectively. Patients were divided into high CD4 and low CD4 group based on CD4+ lymphocytes of 200 and Results After vaccination, CD4+, CD8+ and CD3+ cells increased significantly in all the groups. There was no increase in NK cell activity in patients with high CD4+ lymphocytes and only a marginal increase in patients with low CD4+ lymphocytes (170 to 293/mm3 whereas a marked increase was observed in controls (252 to 490/mm3. After vaccination, although an increase in memory cells was observed in HIV positive patients, yet HBsAb levels were significantly lower than controls (P 4+ group: 8834 mIU/ml, low CD4+ group: 462 mIU/ml Vs. Controls: 16,906 mIU/ml. IL-4 and IL-10 were low in patients. Conclusion Despite a double dose in patients, IL-4 and IL-10, which regulate antibody response, were also lower in patients, and this together with low CD4+ counts and lack of T help, accounted for low HBsAb levels. Vaccination in patients with CD4+ lymphocytes 3 was ineffective. Thus early immunization is advocated in all HIV positive patients at a stage when they are still capable of mounting an adequate immune response

  17. Improving Antigenicity of the Recombinant Hepatitis C Virus Core Protein via Random Mutagenesis

    OpenAIRE

    Chen-Ji Huang; Hwei-Ling Peng; Chih-Yu Cheng

    2011-01-01

    In order to enhance the sensitivity of diagnosis, a recombinant clone containing domain I of HCV core (amino acid residues 1 to 123) was subjected to random mutagenesis. Five mutants with higher sensitivity were obtained by colony screening of 616 mutants using reverse ELISA. Sequence analysis of these mutants revealed alterations focusing on W84, P95, P110, or V129. The inclusion bodies of these recombinant proteins overexpressed in E. coli BL21(DE3) were subsequently dissolved using 6 M ure...

  18. Evaluation of Serum Anti-Hbs Concentration in Children Vaccinated with Recombinant Hepatitis B Vaccine at Birth

    Directory of Open Access Journals (Sweden)

    M Nejad-Ghaderi

    2006-07-01

    Full Text Available Introduction: Vaccination with the major surface antigen of hepatitis B virus (HBsAg induces anti-HBs antibody production and level of 10 IU/L is considered protective. It has been shown that the level of anti-HBs antibody does wane after vaccination. The aim of this study was to evaluate the persistence of anti-HBs antibodies in healthy Iranian children 10 years after primary vaccination. Methods: Blood samples were collected from 146 children, 10 years after completion of primary hepatitis B vaccination course at birth. The sera were tested for anti-HBs, antibody to hepatitis B core antigen (anti-HBc and HBsAg by use of ELISA technique. Results: At 10 years after primary vaccination, 70 (47.9% children had protective levels of antibody (anti-HBs> 10 IU/L with mean titer of 68.1 IU/ml. Moreover, 45 (30.82% children were negative for anti-HBs antibody. Distribution of children according to anti-HBs concentration revealed that the proportion of subjects with antibody titer of 0-10 IU/L, 10-100 IU/L, 100-500 IU/L and 500-1000 IU/L was 52.1%, 24.6%, 20.5% and 2.7%, respectively. All children were negative for HBsAg, although anti-HBc was positive in 11 (7.5% children. There was no difference in the seroprotection rates of males and females. Conclusion: The results of present study show that after 10 years after primary vaccination with recombinant HB vaccine, 47.9% of the children had protective levels of anti-HBs antibody. On basis of the HBsAg and anti-HBc results, it seems that effective immunological memory exists in children. Additional follow-up studies need to be conducted to determine the duration of protection.

  19. Immunogenicity of recombinant hepatitis B virus vaccine in patients with and without chronic hepatitis C virus infection:A case-control study

    Institute of Scientific and Technical Information of China (English)

    Naser Ebrahimi Daryani; Mohsen Nassiri-Toosi; Armin Rashidi; Iman Khodarahmi

    2007-01-01

    AIM: To compare the response of standard hepatitis B virus (HBV) vaccination between patients with chronic hepatitis C virus (HCV) infection and healthy individuals.METHODS: This is a prospective case-control study.A total of 38 patients with chronic HCV infection and 40 healthy controls were included. Vaccination was performed by injection of 20 μg recombinant HBsAg into the deltoid muscle at mo 0, 1 and 6. Anti-HBs concentration was determined 3 mo after the last dose and compared between the two groups. The response pattern was characterized as (1) high-response when the anti-HBs antibody titer was > 100 IU/L, (2) low-response when the titer was 10-100 IU/L and (3) no-response when the titer was < 10 IU/L.RESULTS: In the patient group, there were 10/38(26.3%) non-responders, 8/38 (21.1%)low-responders and 20/38 (52.6%) high-responders. The corresponding values in the control group were 2/40 (5.0%),7/40 (17.5%) and 31/40 (77.5%), respectively. The response pattern was statistically different between the two groups. In multivariate analysis, smoking was a significant confounder, while HCV infection lost its significant correlation with lower antibody response.CONCLUSION: Patients with chronic HCV infection tend to respond weakly to HBV vaccination compared to healthy individuals, though this correlation is not independent according to multivariate analysis.

  20. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    1992-01-01

    920691 The determination of serum hepa-titis B virus DNA by polymerase chain rea-ction in hepatitis B patients treated withalpha-interferon. XU. Jianye(徐建业), et al.Centr Lab, Chongqing Cancer Instit, 630030.Chin J Intern Med, 1992; 31(5): 278-280. To clarify the status of HBV in serum of

  1. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    1997-01-01

    970349 Primary structure and variability of partialsequences in nonstructural gene 5 region of hepatitis Gvirus, CHANG Jinhong(常锦红), et al. Hepatol Instis,People’s Hosp, Beijing Med Univ, Beijing, 100044. NatlMed J China 1997; 77(3): 178-182. Objective: To sequence partial genome of hepatitis G

  2. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    2009209 Effects of chronic hepatitis B virus infection on human hepatic cytochrome P450 2C9.ZHO Fuping(周福平),et al.Dept Infect Dis,Shanghai Changzheng Hosp,Shanghai 200003.Chin J Infect Dis,2009;27(2):94-98.

  3. TH1 and TH2 responses are influenced by HLA antigens in healthy neonates vaccinated with recombinant hepatitis B vaccine.

    Directory of Open Access Journals (Sweden)

    Abdollah Jafarzadeh

    2012-12-01

    Full Text Available The immune response to hepatitis B surface antigen (HBsAg is influenced by several factors, of which HLA antigens and balanced secretion of Th1/Th2 cytokines play important roles. The aim of this study was to evaluate the influence of HLA antigens on cytokine secretion by HBsAg-stimulated peripheral blood mononuclear cells (PBMC from healthy neonates vaccinated with recombinant HBsAg. PBMCs were isolated from 48 Iranian neonates vaccinated with a recombinant HBV vaccine. The cells were stimulated in vitro with rHBsAg and the concentration of IL-4, IL-10, IL-12 and IFN-γ were quantitated in culture supernatant by sandwich ELISA. HLA typing was performed by microlymphocytotoxicity method. Significant diminished secretion of both Th1 (IFN-γ and Th2 (IL-4, IL-10 cytokines was observed in HBsAg-stimulated PBMC from vaccinees expressing the HLA-DR7 compared to DR7 negative vaccinees. Similarly, lower production of these cytokines was also observed in vaccinees with DR7-DR53-DQ2, B7-DR7-DR53-DQ2 and A2-DR7-DR53-DQ2 haplotypes (p<0.05, p <0.005. While HBsAg-stimulated PBMC of DR13+ subjects produced lower levels of Th2-type cytokines (IL-4 and IL-10, those of HLA-B8+ or HLA-A9+ subjects produced higher levels of Th2-type cytokines. Cytokine secretion in response to PHA mitogen was not associated with a given HLA antigen or haplotype and was similarly represented in all groups of subjects irrespective of their HLA complex. These results indicate that HLA antigens may differentially influence cytokine secretion by HBsAg-specific T-cells of healthy neonates vaccinated with recombinant HB vaccine. This phenomenon may have an important implication for control of the immune response to HBsAg vaccine.

  4. Efficacy and safety of triple therapy with recombinant interleukin-1β, recombinant interferon-α and ribavirin in patients with chronic hepatitis C, genotype 1 infection, the non-responders to previous treatment with interferon and ribavirin

    Directory of Open Access Journals (Sweden)

    Yu. V. Lobzin

    2012-01-01

    Full Text Available To assess the efficacy and safety of an experimental course of antiviral therapy with recombinant IL-1β in combination with recombinant interferon-α2b and ribavirin in patients with chronic hepatitis C infected with genotype 1 hepatitis C virus in lacking the virologic response to previous dual therapy pegIFN-α or standard IFN-α and ribavirin has been formed by a group of 50 patients. All patients were treated with recombinant IL-1β – 10 subcutaneous injection at a dose of 0.005 mg/kg every other day, the course of 3 weeks, with 5 courses, recombinant IFN-α2b – subcutaneous injections of 3 million IU every other day for 48 weeks. Ribavirin 1000–1200 mg per day (depending on body weight for 48 weeks. The duration of follow-up after the end of therapy was 24 weeks. Estimated sustained virologic response (SVR. Comparison of SVR according to the version of virologic response in primary treatment showed that the use of the pilot treatment scheme is most effective at relapse (63% than in the «null-responders» (33% and partial (23% responses to the preceding treatment. In the present study did not reveal a single case of serious adverse events or unexpected adverse events. Thus, the inclusion of proven safety of recombinant IL-1β in the scheme of antiviral therapy in combination with standard interferon-α and ribavirin in patients with chronic hepatitis C, and demonstrated the effectiveness of using recombinant IL-1β (in combination with standard interferon-α and ribavirin, especially in patients with recurrent HCV-infection.

  5. Pre-Clinical Evaluation of a Novel Nanoemulsion-Based Hepatitis B Mucosal Vaccine

    OpenAIRE

    Makidon, Paul E.; Bielinska, Anna U.; Nigavekar, Shraddha S.; Janczak, Katarzyna W.; Jessica Knowlton; Alison J Scott; Nicholas Mank; Zhengyi Cao; Sivaprakash Rathinavelu; Michael R Beer; J Erby Wilkinson; Blanco, Luz P.; Jeffrey J Landers; Baker, James R.

    2008-01-01

    BACKGROUND: Hepatitis B virus infection remains an important global health concern despite the availability of safe and effective prophylactic vaccines. Limitations to these vaccines include requirement for refrigeration and three immunizations thereby restricting use in the developing world. A new nasal hepatitis B vaccine composed of recombinant hepatitis B surface antigen (HBsAg) in a novel nanoemulsion (NE) adjuvant (HBsAg-NE) could be effective with fewer administrations. METHODOLOGY AND...

  6. "PERSISTENCE OF ANTI-HBs ANTIBODIES IN HEALTHY IRANIAN CHILDREN VACCINATED WITH RECOMBINANT HEPATITIS B VACCINE AND RESPONSE TO A BOOSTER DOSE"

    Directory of Open Access Journals (Sweden)

    A. Jafarzadeh

    2005-05-01

    Full Text Available Long-term protection against hepatitis B virus (HBV is dependent on persistence of anti-HBs antibodies and/or strong immunological memory. In this study we evaluated the persistence of anti-HBs antibodies in healthy Iranian children 5 years after primary vaccination and the response to a booster dose using recombinant hepatitis B vaccine. Totally, 81 children who had received primary course of hepatitis B vaccine at 0, 1.5 and 9 months of age were included in this study. A booster dose of hepatitis B vaccine was administered at 5 years after completion of primary vaccination program. Children were tested for anti-HBs antibody just before administration of booster dose and at 4 weeks after booster vaccination. An 81.5% seroprotection rate (anti-HBs > 10 IU/L was observed 5 years after primary vaccination. After administration of booster dose, 100% of the children developed protective level of anti-HBs antibody and geometric mean titer rose from 206 IU/L to 1278 IU/L. These results indicate the existence of an effective immunological memory over a period of 5 years after primary vaccination with recombinant hepatitis B vaccine in healthy Iranian children.

  7. Immunogenicity of recombinant hepatitis B vaccine: comparison of two different vaccination schedules

    OpenAIRE

    Agladioglu, S.; Beyazova, U.; Camurdan, A. D.; Sahin, F.; Atak, A.

    2010-01-01

    Background Neonatal immunization with hepatitis B (HB) vaccine induces protective levels of antibody (anti-HBs ≥10 IU/L) in a majority of vaccines. However, the duration of protection after HB vaccination in infants is unknown. A smaller proportion of children vaccinated beginning at birth with three doses of HB vaccine were found to have protective titers 5–10 years after initial vaccination. Long-term efficacy of HB vaccine depends mainly on peak antibody levels after vaccination, and subje...

  8. Inhibition on the production of collagen type Ⅰ, Ⅲ of activated hepatic stellate cells by antisense TIMP-1 recombinant plasmid

    Institute of Scientific and Technical Information of China (English)

    Wen-Bin Liu; Chang-Qing Yang; Wei Jiang; Yi-Qing Wang; Jing-Sheng Guo; Bo-Ming He; Ji-Yao Wang

    2003-01-01

    AIM: To investigate the inhibition effects on the productionof collagen type I, Ⅲ secreted by activated rat hepatic stellatecells (rHSCs) by antisense tissue inhibitors of metalloproteinase1 (TIMP-1) recombinant plasmid through elevating interstitialcollagenase activity.METHODS: rHSCs were extracted from normal rat liverby pronase and collagenase digestion and purified bycentrifugal elutriation, and were cultured on plastic dishesuntil they were activated to a myofibroblastic phenotypeafter 7-10 days. RT-Nest-PCR and gene recombinanttechniques were used to construct the rat antisense TIMP-1 recombinant plasmids which can express in eucaryoticcells. The recombinant plasmid and the pcDNA3 emptyplasmid were transfected in rHSCs by Effectene (QIAGEN)separately. Cells were selected after growing in DMEMcontaining 400 μg/ml G418 for 2-3 weeks. Expression ofexogenous gene was assessed by Northern blot, andexpression oflIMP-1 in rHSCs was determined by Northernblot and Western blot. We tested the interstitial collagenaseactivity with FITC-labled type I collagen as substrate.Ultimately, we quantified the type Ⅰ, Ⅲ collagen byWestern blot.RESULTS: The exogenous antisense TIMP-1 recombinantplasmid could be expressed in rHSCs well, which couldblock the expression of TIMP-1 greatly, the ratio of TIMP-1/GAPDH was 0.67, 2.41, and 2.97 separately at mRNAlevel (P<0.05); the ratio of TIMP-1/β-actin was 0.31, 0.98and 1.32 separately at protein level (P<0.05); It mightelevate active and latent interstitial collagenase activity,the collagenase activity was 0.3049, 0.1411 and 0.1196respectively. (P<0.05), which led to promotion thedegradation of type Ⅰ, Ⅲ collagen, the ratio of collagen I/β-actin was 0.63, 1.78 and 1.92 separately (P<0.05); andthe ratio of collagen Ⅲ/β-actin was 0.59, 1.81 and 1.98separately (P<0.05).CONCLUSION: These data shows that the antisense TIMP-1 recombinant plasmid has the inhibitory effects on theproduction of type Ⅰ, Ⅲ collagens

  9. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    2005226 Characteristics of peripheral blood T lymphocyte subsets in hepatitis B patients. FAN Zhen-ping(范振平),et al. Center Bio Ther, Instit Infect Dis, 302 Hosp Chin PLA, Beijing 100039. World Chin J Digestol, 2005;13(2): 194-197. Objective: To characterize the T-lymphocyte subsets in peripheral blood of patients with acute and chronic hepatitis B, and to explore their relations with the disease state. Methods: Peripheral blood

  10. Controlled trial of immune response of preterm infants to recombinant hepatitis B and inactivated poliovirus vaccines administered simultaneously shortly after birth

    OpenAIRE

    Linder, N.; Handsher, R.; German, B.; Sirota, L.; Bachman, M.; Zinger, S.; Mendelson, E.; Barzilai, A.

    2000-01-01

    AIM—The study was conducted to evaluate the immunogenicity of an early, extra dose of enhanced inactivated poliovirus vaccine (IPV) administered simultaneously with recombinant hepatitis B vaccine (HBV) to preterm infants shortly after birth.
METHODS—Three groups were studied. Fifty preterm infants received IPV intramuscularly within 24 hours of birth, in addition to routine recommended childhood immunisations. Fifty two preterm infants and 35 full term infants receive...

  11. High expression of hepatitis B virus based vector with reporter gene in hepatitis B virus infection system

    Institute of Scientific and Technical Information of China (English)

    Shi-Hong Li; Wen-Ge Huang; Bing Huang; Xi-Gu Chen

    2007-01-01

    AIM: To construct a hepatitis B virus (HBV)-based vector with a reporter gene and to establish an HBV infection system to evaluate the availability of the vector.METHODS: The HBV-based vectors with green fluorescence protein (GFP) were packaged into the liver of immunodeficient mice through transfer and helper plasmid using hydrodynamic technology. Wild type HBV (wt HBV) was provided by plasmid MC2009. Primary human hepatocytes (PHH) were isolated and infected with recombinant HBV (rHBV) or wt HBV. GFP expression was monitored by confocal and flow cytometry. HBV DNA and HBV surface antigen (HBSAg) were analyzed by PCR and ELISA.RESULTS: 3 × 107 wt HBV copies/mL and 5 × 106 rHBV copies/mL were collected from mice serum. In the wt HBV infected group, HBV progeny was 2 × 107 copies/mL and HBSAg was 770 ng/mL. In the rHBV infected group, GFP fluorescence was detected on d 3 post-infection and over 85% of the parenchymal cells expressed green fluorescence on d 12 post-infection. Compared with wt HBV in the PHH infection system, no rHBV DNA or HBSAg were detected in PHH culture media.CONCLUSION: An effective HBV based vector was developed, which proved to be a useful HBV infection system. This vector and infection system can be applied to develop a therapeutic vector and study the HBV life cycle and viral pathogenesis.

  12. Heads or Tails: Genotyping of Hepatitis C Virus Concerning the 2k/1b Circulating Recombinant Form

    Science.gov (United States)

    Schuermans, Wim; Orlent, Hans; Desombere, Isabelle; Descheemaeker, Patrick; Van Vlierberghe, Hans; Geerts, Anja; Verhelst, Xavier; Reynders, Marijke; Padalko, Elizaveta

    2016-01-01

    As different hepatitis C virus (HCV) genotypes respond differently to initiated therapy, correct HCV genotyping is essential. A potential risk for misclassification of the intergenotypic HCV circulating recombinant form (CRF) 2k/1b strains exists, depending on the genotyping method used. The aim was to investigate the differences in HCV genotyping methods with regard to CRF 2k/1b and to gain insight in the prevalence of the CRF 2k/1b. Genotyping results by Versant HCV Genotype Assay were compared with nonstructural protein 5B (NS5B) sequencing. In total, from November 2001 until March 2015, 3296 serum samples were analyzed by Versant HCV Genotype Assay. As misclassified CRF is harbored among HCV genotype 2, we further focused our search on 142 (4.3%) samples positive for HCV genotype 2. On 116 (81.7%) retrieved samples, the NS5B sequencing was performed. Twelve out of the 116 retrieved samples (10.3%) were classified as CRF 2k/1b by sequencing of the NS5B region. Ten of these 12 samples were originally misclassified as genotype 2a or 2c, while 2 of them were misclassified as genotype 2. Our results show that the current prevalence of CRF 2k/1b is underestimated. The importance of correct HCV genotyping is emphasized, considering the tailored choice of treatment regimen and overall prognosis. PMID:27563879

  13. PRELIMINARY REPORT OF THE USE ON ADULTS OF A RECOMBINANT YEAST-DERIVED HEPATITIS B VACCINE MANUFACTURED BY INSTITUTO BUTANTAN

    Directory of Open Access Journals (Sweden)

    Angela Aparecida COSTA

    1997-01-01

    Full Text Available Three 10 µg doses of the recombinant hepatitis B vaccine, manufactured by Instituto Butantan by original technology, were administered in a adult population, mean age 30 years old, following the 0, 1 and 6 months schedule immunization. The clinical trial was considered satisfactory in terms of immunogenicity (anti-HBs titers between 17.5-29500 IU/l, seroconversion 95.3% and reactogenicity (no incapacitating side effectsTrês doses de 10 µg da vacina recombinante contra hepatite B, produzida pelo Instituto Butantan, através de tecnologia própria, foram administradas numa população de adultos (idade média 30 anos, seguindo o esquema de imunização 0, 1 e 6 meses após a primeira dose. A avaliação clínica da vacina foi considerada satisfatória em termos de imunogenicidade (títulos dos anticorpos anti-HBs entre 17,5-29500 UI/1, soroconversão 95,3% e reatividade (sem efeitos colaterais e sintomas clínicos relevantes

  14. Comparison of three different recombinant hepatitis B vaccines: GeneVac-B, Engerix B and Shanvac B in high risk infants born to HBsAg positive mothers in India

    OpenAIRE

    Velu, Vijayakumar; Nandakumar, Subhadra; Shanmugam, Saravanan; Jadhav, Suresh Sakharam; Kulkarni, Prasad Suryakant; Thyagarajan, Sadras Panchatcharam

    2007-01-01

    AIM: To evaluate a low cost Indian recombinant hepatitis B vaccine GeneVac-B® for its immunogenicity and safety in comparison to Engerix B® and Shanvac B® vaccine in high risk newborn infants born to hepatitis B surface antigen (HBsAg) positive mothers.

  15. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008312 Impact of hepatitis B virus infection on the activity of hematopoietic stem cell.SHI Yanmei(石雁梅),et al.Dept Infect Dis,1st Clin Coll,Harbin Med Univ,Harbin 150001.Chin J Infect Dis 2008;26(4):197-201.Objective To study the impact of hepatitis B virus (HBV)infection on the activity of cord hematopoieticstem cells.Methods CD34+cells were isolated from healthy human cord blood by mini MACS.Cells were

  16. Immunogenicity of Hepavax-Gene recombinant hepatitis B vaccine in INML workers, Colombia.

    Directory of Open Access Journals (Sweden)

    Heber Siachoque

    2009-11-01

    Full Text Available Objetivo: Evaluar la inmunogenicidad de la vacuna recombinante Hepavax-Gene para hepatitis B desde la última dosis administrada en trabajadores del INML. Materiales y métodos: Estudio de corte transversal en 603 trabajadores de la salud con mínimo 3 dosis de vacuna recombinante (0, 1, 6 meses donde se midieron los niveles de anticuerpos anti-HBs con la técnica de ELISA entre diciembre de 2000 y enero de 2001 desde la aplicación de la última dosis de la vacuna, que varió entre 1 y 6 años. Resultados: El grupo de estudio lo conformaron 344 hombres y 259 mujeres, con un promedio de edad de 38.8±7.3 años. El nivel de protección fue 90.7% (>10 U/l que disminuyó significativamente con el tiempo de aplicación de la última dosis de la vacuna (p45 (RRI=3.58, IC 95%:1.83, 6.99 con respecto a Conclusión: La vacuna recombinante Hepavax-Gene anti-HBs tiene alta efectividad en los trabajadores de la salud (90.7% aunque presenta disminución de protección a mayor tiempo de aplicación de la última dosis y al aumentar la edad del trabajador.

  17. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008449 A cross-sectional survey of occult hepatitis B virus infection in HIV-infected patients. MA Jianxin(马建新), et al.Dept Infect Dis, Shanghai Public Health Clin Center, Shanghai 201508. Chin J Intern Med 2008;47(7):574-577. Objective To assess the prevalence of occult HBV infection in HIV-infected patients.

  18. Diagnosis of hepatic fibrosis in hepatitis B patients by logistic regression modeling based on plasma amino acid ratio and age

    Institute of Scientific and Technical Information of China (English)

    张占卿

    2013-01-01

    Objective To explore the efficacy of logistic regression modeling based on plasma amino acid profile and patient age,for diagnosing hepatic fibrosis in patients with chronic hepatitis B (CHB) .Methods One-hundredand-forty-eight patients (108 males;mean age:38.1±11.9 years,range:16—72 years) histologically

  19. Characterization of hepatitis C virus recombinants with chimeric E1/E2 envelope proteins and identification of single amino acids in the E2 stem region important for entry

    DEFF Research Database (Denmark)

    Carlsen, Thomas H R; Scheel, Troels K H; Ramirez, Santseharay;

    2013-01-01

    The hepatitis C virus (HCV) envelope proteins E1 and E2 play a key role in host cell entry and represent important targets for vaccine and drug development. Here, we characterized HCV recombinants with chimeric E1/E2 complexes in vitro. Using genotype 1a/2a JFH1-based recombinants expressing 1a...... core-NS2, we exchanged E2 with functional isolate sequences of genotypes 1a (alternative isolate), 1b, and 2a. While the 1a-E2 exchange did not impact virus viability, the 2a-E2 recombinant was nonviable. After E2 exchange from three 1b isolates, long delays were observed before spread of infection....... For recovered 1b-E2 recombinants, single E2 stem region amino acid changes were identified at residues 706, 707, and 710. In reverse genetic studies, these mutations increased infectivity titers by ~100-fold, apparently without influencing particle stability or cell binding although introducing slight decrease...

  20. Preclinical evaluation of a two-dose vaccination schedule of recombinant Hansenula Polymorpha hepatitis B vaccine in animals

    OpenAIRE

    Li, Jin; Zhang, Deyou; Ma, Rui; Yang, Xuqin; Wang, Xi; Li, Caimei; Zhang, Sucai; Xue, Hesheng; Zhao, Kai; Zhuang, Hui

    2013-01-01

    Aims: The current 3-dose regimen of hepatitis B vaccination for infants requiring over 6 mo period may pose the poor rate of compliance and later protection from hepatitis B virus (HBV) infection. This preclinical study is to investigate the feasibility of reducing the number of doses of hepatitis B (HB) vaccine.

  1. Ef ifcacy of Shanvac-B recombinant DNA hepatitis B vaccine in health care workers of Nor thern India

    Institute of Scientific and Technical Information of China (English)

    Varsha Thakur; Nirupma T Pati; Rajkumar C Gupta; Shiv K Sarin

    2010-01-01

    BACKGROUND: Health care workers (HCWs) constitute a high-risk population of HBV infection. There are limited data on the efifcacy of vaccination in HCWs in India. This study was to evaluate the efifcacy of indigenous recombinant hepatitis B vaccine, Shanvac-B, in HCWs. METHODS: In 597 HCWs screened before the vaccination, 216 (36.2%) showed the presence of at least one of the markers of HBV/HCV infection. Of the remaining 381 (63.8%) HCWs who were considered for vaccination, only 153 (age 18-45 years; 48 males and 105 females) were available for ifnal assessment. These HCWs received 20 μg of vaccine at 0, 1 and 6 months. They were asked for the reactogenicity and monitored for the seroprotective efifcacy of the vaccination. Anti-HBs titres were measured after vaccination at 1, 2 and 7 months. The presence of anti-HBs titers equal to 1 MIU/ml was considered as seroconversion and that of titres greater than 10 MIU/ml as seroprotection. RESULTS:After vaccination, 32 males (67%) and 76 females (72%) showed seroconvertion;ifnally 12 (25%) of the males and 47 (45%) of the females were seroprotected. Seroprotection at 2 and 7 months was more dominant in the females than in the males (96% vs. 56%, P=0.001, 100% vs. 85%, P=0.0001), respectively. Geometric mean titres of anti-HBs after vaccination were also higher in the females than in the males (257±19.7 vs. 29±1.88 MIU/ml, P=0.01, 1802±35.2 vs. 306±13.6 MIU/ml, P≤0.05, 6465±72 vs. 2142±73.6 MIU/ml, P CONCLUSIONS:The presence of HBsAg in HCWs indicates that a high proportion of HCWs are infected with HBV and HCV in India. Recombinant indigenous vaccine Shanvac-B is highly efifcacious in HCWs, and its immunogenicity is signiifcantly higher in females than in males. However, pre-vaccination screening of HCWs is strongly recommended in India.

  2. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    2008079 Relationship of HBV genotype and bcp and pc mutations with HBV DNA rebound after lamivudine therapy. SU Minghua(苏明华), et al. Dept Infect Dis Clin Hosp, Guangxi Med Univ, Nanning 530027. World Chin J Digestol 2007;15(33):3507-3513. Objective To investigate the relationship of HBV gene mutations with HBV DNA rebound after lamivudine therapy. Methods Twenty-seven hepatitis B patients with HBV DNA rebound after

  3. Species association of hepatitis B virus (HBV in non-human apes; evidence for recombination between gorilla and chimpanzee variants.

    Directory of Open Access Journals (Sweden)

    Sinéad Lyons

    Full Text Available Hepatitis B virus (HBV infections are widely distributed in humans, infecting approximately one third of the world's population. HBV variants have also been detected and genetically characterised from Old World apes; Gorilla gorilla (gorilla, Pan troglodytes (chimpanzee, Pongo pygmaeus (orang-utan, Nomascus nastusus and Hylobates pileatus (gibbons and from the New World monkey, Lagothrix lagotricha (woolly monkey. To investigate species-specificity and potential for cross species transmission of HBV between sympatric species of apes (such as gorillas and chimpanzees in Central Africa or between humans and chimpanzees or gorillas, variants of HBV infecting captive wild-born non-human primates were genetically characterised. 9 of 62 chimpanzees (11.3% and two from 11 gorillas (18% were HBV-infected (15% combined frequency, while other Old world monkey species were negative. Complete genome sequences were obtained from six of the infected chimpanzee and both gorillas; those from P. t .ellioti grouped with previously characterised variants from this subspecies. However, variants recovered from P. t. troglodytes HBV variants also grouped within this clade, indicative of transmission between sub-species, forming a paraphyletic clade. The two gorilla viruses were phylogenetically distinct from chimpanzee and human variants although one showed evidence for a recombination event with a P.t.e.-derived HBV variant in the partial X and core gene region. Both of these observations provide evidence for circulation of HBV between different species and sub-species of non-human primates, a conclusion that differs from the hypothesis if of strict host specificity of HBV genotypes.

  4. Rituximab-Based Treatment, HCV Replication, and Hepatic Flares

    Directory of Open Access Journals (Sweden)

    Evangelista Sagnelli

    2012-01-01

    Full Text Available Rituximab, a chimeric mouse-human monoclonal antibody directed to the CD20 antigen expressed on pre-B lymphocytes and mature lymphocytes, causes a profound B-cell depletion. Due to its peculiar characteristics, this drug has been used to treat oncohaematological diseases, B cell-related autoimmune diseases, rheumatoid arthritis, and, more recently, HCV-associated mixed cryoglobulinaemic vasculitis. Rituximab-based treatment, however, may induce an increased replication of several viruses such as hepatitis B virus, cytomegalovirus, varicella-zoster virus, echovirus, and parvovirus B19. Recent data suggest that rituximab-based chemotherapy induces an increase in HCV expression in hepatic cells, which may become a target for a cell-mediated immune reaction after the withdrawal of treatment and the restoration of the immune control. Only a few small studies have investigated the occurrence of HCV reactivation and an associated hepatic flare in patients with oncohaematological diseases receiving R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone. These studies suggest that the hepatic flares are frequently asymptomatic, but life-threatening liver failure occurs in nearly 10% of cases.

  5. Hepatitis

    Institute of Scientific and Technical Information of China (English)

    1995-01-01

    950335 Preliminary study on the treatment of chil-dren HBV associated glomerulonephritis with humanrecombinant α1 interferon(rhiFNα1).FANG Lijun(方利君),et al.Pediatr Hosp,Shanghai Med Univ,Shang-hai,200032.Chin J Nephrol 1994;10(6):329-331.Three cases of HBV associated glomerulonephritis(HBV-GN) are treated by human recombinant inter-feron (rhIFNα1) made in China.All of these cases havemanifestation of nephrotic syndrome.Their clinicalcharacteristics,pathological findings and molecular

  6. Enhanced specific immune responses by CpG DNA in mice immunized with recombinant hepatitis B surface antigen and HB vaccine

    OpenAIRE

    Wang Xingtai; Hu Zhongyu; He Peng; Zhang Xiancheng; Liang Zhenglun

    2011-01-01

    Abstract Background Hepatitis B vaccine adjuvant, alum, is generally used for vaccination although it does not stimulate Th1 immunity and 10% of the population has low or no antibody response. Efforts have been continued to find more efficient vaccine adjuvants for better antibody response as well as stimulation of Th1 immunity. Methods CpG DNA was used as an adjuvant for recombinant HBsAg to immunize 6- to 8-week-old female BALB/c mice with or without alum for different dosages. The producti...

  7. Functional analyses of GB virus B p13 protein: development of a recombinant GB virus B hepatitis virus with a p7 protein

    DEFF Research Database (Denmark)

    Takikawa, Shingo; Engle, Ronald E; Emerson, Suzanne U;

    2006-01-01

    plus part of p7) was nonviable. However, a mutant lacking amino acid 614-669 (p6) produced high titer viremia andacute resolving hepatitis; viruses recovered from both animals lacked the deleted sequence and had no other mutations. Thus, p6 was dispensable but p7 was essential for infectivity. The...... availability of a recombinant GBV-B virus containing a p7 protein with similarities to the HCV p7 will enhance the relevance of this model and will be of importance for identifying compounds that inhibit p7 function as additional therapeutic agents....

  8. Combination treatment with hepatitis C virus protease and NS5A inhibitors is effective against recombinant genotype 1a, 2a, and 3a viruses

    DEFF Research Database (Denmark)

    Gottwein, Judith M; Jensen, Sanne B; Li, Yi-Ping;

    2013-01-01

    With the development of directly acting antivirals, hepatitis C virus (HCV) therapy entered a new era. However, rapid selection of resistance mutations necessitates combination therapy. To study combination therapy in infectious culture systems, we aimed at developing HCV semi-full-length (semi...... against previously developed 2a recombinants J6/JFH1 and J6cc. Daclatasvir had intermediate efficacy against J6/JFH1, while low sensitivity was confirmed against J6cc. Using a cross-titration scheme, infected cultures were treated until viral escape or on-treatment virologic suppression occurred. Compared...

  9. Prevalence of hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis D virus and hepatitis E virus as causes of acute viral hepatitis in North India: A hospital based study

    OpenAIRE

    Jain, P; Prakash, S.; Gupta, S; Singh, K.P.; Shrivastava, S; Singh, D. D.; Singh, J; Jain, A.

    2013-01-01

    Context: Acute viral hepatitis (AVH) is a major public health problem and is an important cause of morbidity and mortality. Aim: The aim of the present study is to determine the prevalence of hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV) and hepatitis E virus (HEV) as causes of AVH in a tertiary care hospital of North India. Settings and Design: Blood samples and clinical information was collected from cases of AVH referred to the Grade I v...

  10. Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells

    Institute of Scientific and Technical Information of China (English)

    CHEN Yong-bing; SHI Xian-jie; LU Gang; NIE Hong-feng; SHEN Xiao-qing; YU Cong-hui; GONG Jian-ping

    2011-01-01

    Background Indoleamine-2,3-dioxygenase (IDO) is proven to suppress hepatitis B virus (HBV) specific immune response and depletion of IDO may be a useful approach for HBV therapy. To test this concept, we constructed recombinant adenovirus with human IDO and HBV preS, which would form the basis for future in vivo experiments.Methods The fragment of human IDO and HBV preS cDNA were subcloned into multiple cloning sites in an adenoviral vector system containing two cytomegalovirus (CMV) promoters. Recombination was conducted in the Escherichia coli BJ5183. The recombinant adenovirus containing hlDO gene and HBVpreS gene was packaged and amplified in 293 cells.Integration was confirmed by polymerase chain reaction as well as the quantification of viral titers. HepG2 cells were infected with the recombinant adenovirus and mRNA and protein specific for hlDO and HBVpreS was detected by RT-PCR and Western blotting respectively.Results The recombinant adenovirus was produced successfully. Its titer was 2.5x109 efu/ml. IDO and HBVpreS mRNA as well as the encoded proteins could be found in transfected HepG2 cells, but not in control HepG2 cells.Conclusion The transfer of hlDO-HBVpreS with double-promoter adenoviral vector was efficient. The recombinant adenovirus with hlDO and HBVpreS would provide the experimental basis for future studies.

  11. Hepatitis B surface antigenemia following recombinant Engerix B hepatitis B vaccine in an 81-year-old ESRD patient on hemodialysis.

    Science.gov (United States)

    Onuigbo, Macaulay A C; Nesbit, Ashley; Weisenbeck, Jacquelyn; Hurlburt, Jill

    2010-05-01

    The first cases of transient hepatitis B surface antigenemia (HBsAg) in adults following hepatitis B virus (HBV) immunization were reported in the 1990s. HBV immunization is mandatory for all hemodialysis (HD) patients. Ly et al. who demonstrated transient HBsAg in eight out of nine HD patients following HBV vaccine concluded that HD patients should not be screened for HBV within a week of HBV immunization and that positive HBsAg within a month of HBV immunization must be interpreted with caution. We present an 81-year-old woman on HD, who needed a booster Recombivax (Merck, Whitehouse Station, NJ, USA) vaccine after remaining hepatitis B surface antibody (HBsAb) negative from previous vaccinations. The HD Unit had switched to Engerix B (GlaxoSmithKline, Atlanta, GA, USA) HBV vaccine. Two days after the first Engerix B vaccine, HBsAg was detected. She was asymptomatic; ALT was 25 U/L. Repeat testing for HBsAg, HBsAb, hepatitis B E antigen (HB E Ag), and hepatitis B DNA (HB DNA), a week later, all returned negative. Previous reports of transient HBsAg following HBV vaccines were after Engerix B vaccination. Our patient is unusual since she had received both brands of HBV vaccines, sequentially, at different times. Twice, HBsAg tests completed as early as 5 days following Recombivax vaccine were negative. We submit that positive HBsAg tests are more likely following Engerix B vaccines. We reemphasize previous recommendations that patients should not be screened for HBsAg HBV immunization. This is particularly important in HD units where hepatitis B screening is carried out routinely all year round and hepatitis B vaccinations are commonplace. Very strict schedules must be adopted to avoid false positive HBsAg tests. PMID:20446799

  12. Optimizing Interferon Alfa Based Therapy for Chronic Hepatitis C

    NARCIS (Netherlands)

    R. Roomer (Robert)

    2011-01-01

    textabstractThe hepatitis C virus was first discovered in 1989 as the major cause of chronic non-A non-B hepatitis. The hepatitis C virus is a single stranded RNA virus that belongs to the family of flaviviruses. The primary target of the hepatitis C virus are hepatocytes where viral particles repli

  13. Abrupt onset of type 1 diabetes mellitus during recombinant interferon-alpha 2b therapy in a patient with chronic hepatitis B

    Institute of Scientific and Technical Information of China (English)

    Yuan-Yuan Lv; Bing-Yin Shi; Hui Guo

    2008-01-01

    We describe a case of a 33-year-old female patient with chronic hepatitis B who developed type 1 diabetes mellitus (DM) after a 13-mo period of treatment with recombinant human interferon-alpha (IFN-α) 2b. The patient presented with polydipsia, polyuria, hypergly-cemia, diabetic ketoacidosis, combined with C-peptide secretion deficiency and positive islet cell autoantibody (ICAb). IFN-α 2b treatment was terminated and in-stead insulin treatment was initiated. Five months after cessation of the recombinant human IFN-α 2b therapy,the patient remained insulin-dependent. Her serum HBV DNA became negative and serum transaminase returned to the normal level after a 10-mo period ofIFN therapy. Type 1 DM induced by IFN-α is relativelyrare in patients with chronic hepatitis B. We should paymore attention to patients on IFN-α therapy to avoid destruction of pancreatic beta cells. This is the first case report from China.

  14. Immunocapture Enzyme-Linked Immunosorbent Assay for Assessment of In Vitro Potency of Recombinant Hepatitis B Vaccines▿

    OpenAIRE

    Shanmugham, Rajalakshmi; Thirumeni, Nagarajan; Rao, Varaprasada Sankarashetty; Pitta, Vidyasagar; Kasthuri, Saranyarevathy; Singanallur, Nagendrakumar Balasubramanian; Lingala, Rajendra; Mangamoori, Lakshmi Narsu; Villuppanoor, Srinivasan Alwar

    2010-01-01

    Quantification of hepatitis B surface antigen (HBsAg) or relative in vitro potency in the final vaccines is a prerequisite for hepatitis B vaccine batch release. The commercial kit for automated analysis (AxSYM) is expensive, and an alternative is required for the estimation of HBsAg in hepatitis B vaccines. Mouse monoclonal antibodies (MAbs) specific for HBsAg were developed and characterized. One of the monoclonal antibodies (HBs06) was used in development of an immunocapture ELISA (IC-ELIS...

  15. Transformation of tobacco plant (Nicotiana tabacum L. with the recombinant hepatitis B virus genes 35SHBsAg and 35SHBsAgER

    Directory of Open Access Journals (Sweden)

    Juliana Martins Ribeiro

    2010-03-01

    Full Text Available The recombinant surface antigen of hepatitis B virus (HBsAg, purified from transgenic plants, proved to be efficient when utilized for raising anti-HB antibodies for the prevention of hepatitis B. Because of the important role of the HBsAg antigen in hepatitis B prevention, the coding sequence of HBsAg antigen, with or without the addition of the carboxi-terminus sequence for protein retention in the endoplasmatic reticulum, was linked to cauliflower mosaic virus 35S promoter, tobacco mosaic virus leader sequence Ω, and the transcription terminator sequence. The aim of this work was to clone the chimeric gene 35SHBsAgER in the plant expression vector pGPTV/Kan/Asc. The resulting plasmid, called pG35SHBsAgER, and another plasmid produced previously in our laboratory called pG35SHBsAg, were transferred to Agrobacterium tumefaciens, and tobacco leaves, of the SR1 cultivar were used as explants for genetic transformation. Twenty-one fully regenerated plants were obtained (10 for the pG35SHBsAg construction and 11 for the pG35SHBsAgER construction. The genomic DNA of all plants was analyzed by PCR, and the presence of the transgene was confirmed in all plants.

  16. Construction and Immunogenicity Testing of Whole Recombinant Yeast-Based T-Cell Vaccines.

    Science.gov (United States)

    King, Thomas H; Guo, Zhimin; Hermreck, Melanie; Bellgrau, Donald; Rodell, Timothy C

    2016-01-01

    GlobeImmune's Tarmogen(®) immunotherapy platform utilizes recombinant Saccharomyces cerevisiae yeast as a vaccine vector to deliver heterologous antigens for activation of disease-specific, targeted cellular immunity. The vaccines elicit immune-mediated killing of target cells expressing viral and cancer antigens in vivo via a CD8(+) CTL-mediated mechanism. Tarmogens are not neutralized by host immune responses and can be administered repeatedly to boost antigen-specific immunity. Production of the vaccines yields stable off-the-shelf products that avoid the need for patient-specific manufacturing found with other immunotherapeutic approaches. Tarmogens for the treatment of chronic hepatitis B and C and various cancers were well tolerated and immunogenic in phase 1 and 2 clinical trials encompassing >600 subjects. The platform is being widely utilized in basic vaccine research and the most rapid path to success in these endeavors follows from optimal immunoassay selection and execution. This chapter provides detailed methods for the construction and preclinical immunogenicity testing of yeast-based immunotherapeutic products to support the rapid and efficient use of this versatile technology. PMID:27076321

  17. Immunogenicity of recombinant hepatitis B vaccine in treatment-naïve and treatment-experienced chronic hepatitis C patients: The effect of pegylated interferon plus ribavirin treatment

    OpenAIRE

    Elefsiniotis, Ioannis S; Vezali, Elena; Kamposioras, Konstantinos; Pantazis, Konstantinos D.; Tontorova, Radostina; Ketikoglou, Ioannis; Moulakakis, Antonios; Saroglou, George

    2006-01-01

    AIM: To retrospectively evaluate the vaccination-induced anti-HBs seroconversion rates in treatment-naïve and treatment-experienced chronic hepatitis C (CHC) patients. Also to prospectively evaluate the seroconversion rates in CHC patients during pegylated interferon (PEG) plus ribavirin (RIB) treatment.

  18. Purification of hepatitis B surface antigen virus-like particles from recombinant Pichia pastoris and in vivo analysis of their immunogenic properties.

    Science.gov (United States)

    Gurramkonda, Chandrasekhar; Zahid, Maria; Nemani, Satish Kumar; Adnan, Ahmad; Gudi, Satheesh Kumar; Khanna, Navin; Ebensen, Thomas; Lünsdorf, Heinrich; Guzmán, Carlos A; Rinas, Ursula

    2013-12-01

    Following earlier studies on high-level intracellular production of hepatitis B surface antigen (HBsAg) using recombinant Pichia pastoris, we present here in detail an enhanced method for the purification of recombinant HBsAg virus-like particles (VLPs). We have screened various detergents for their ability to promote the solubilization of recombinant intracellular HBsAg. In addition, we have analyzed the effect of cell disruption and extraction regarding their impact on the release of HBsAg. Our results show that introduction of the mild nonionic detergent Tween 20 in the initial process of cell lysis at ∼600bars by high pressure homogenization leads to the best results. The subsequent purification steps involved polyethylene glycol precipitation of host cell contaminants, hydrophobic adsorption of HBsAg to colloidal silica followed by ion-exchange chromatography and either isopycnic density ultracentrifugation or size exclusion chromatography for the recovery of the VLPs. After final KSCN treatment and dialysis, a total yield of ∼3% with a purity of >99% was reached. The pure protein was characterized by electron microscopy, showing the presence of uniform VLPs which are the pre-requisite for immunogenicity. The intramuscular co-administration of HBsAg VLPs, with either alum or a PEGylated-derivative of the toll-like receptor 2/6 agonist MALP-2, to mice resulted in the elicitation of significantly higher HBsAg-specific IgG titers as well as a stronger cellular immune response compared to mice vaccinated with a gold standard vaccine (Engerix™). These results show that P. pastoris derived HBsAg VLPs exhibit a high potential as a superior biosimilar vaccine against hepatitis B. PMID:24141044

  19. Anti-HBs levels in infants of hepatitis B carrier mothers after delayed active immunization with recombinant vaccine concomitant with DTP-polio vaccine: Is there need for a second dose of HBIg?

    OpenAIRE

    Grosheide, Pia Maria; Del Canho, R.; Voogd, M.; Heijtink, R.A.; Schalm, Solko

    1994-01-01

    textabstractThe need for an additional dose of hepatitis B immune globulin (HBIg) was studied by comparing infants receiving 1 ml HBIg at birth followed by hepatitis B immunization, concomitant with DTP-polio vaccine, at 3, 4, 5 and 11 months (schedule E), with infants receiving the same schedule with additional HBIg at 3 months (schedule F). The immune response to recombinant hepatitis B vaccine (20 μg) was evaluated in 195 infants born to HBsAg-positive mothers allocated to groups E and F a...

  20. Immunogenicity of recombinant hepatitis B vaccine in treatment-naive and treatment-experienced chronic hepatitis C patients: The effect of pegylated interferon plus ribavirin treatment

    Institute of Scientific and Technical Information of China (English)

    Ioannis S Elefsiniotis; Elena Vezali; Konstantinos Kamposioras; Konstantinos D Pantazis; Radostina Tontorova; Ioannis Ketikoglou; Antonios Moulakakis; George Saroglou

    2006-01-01

    AIM: To retrospectively evaluate the vaccinationinduced anti-HBs seroconversion rates in treatmentnaive and treatment-experienced chronic hepatitis C (CHC) patients. Also to prospectively evaluate the seroconversion rates in CHC patients during pegylated interferon (PEG) plus ribavirin (RIB) treatment.METHODS: Seventy treatment-naive CHC patients (group A), 22 sustained virological responders-SVR following interferon (IFN) plus RIB treatment CHC patients (group B) and 121 healthy subjects (group C) had been participated in the same HBV vaccination schedule (20 μg, 0-1-6 mo). Seroconversion was considered if anti-HBs levels were above 10 mIU/mL within 3 mo following the third dose of the vaccine.Moreover, we prospectively selected 30 non-cirrhotic CHC patients and evaluated them for the efficacy of the same vaccine schedule randomizing them in two groups:Group-1, 15 CHC patients received the first dose of the vaccine in parallel with the initiation of PEG plus RIB treatment and Group-2, 15 patients received the same vaccination schedule without concomitant treatment.Determination of anti-HBs was performed at mo 1, 2,and 7. Statistical analysis of data was based on ANOVA student's t-test and chi-square analysis (P < 0.05).RESULTS: Fifty-eight of 70 group A patients (82.85%),20/22 group B (90.9%) and 112/121 healthy subjects (92.56%) had been seroconverted. The seroconversion rates were significantly higher in the control group than in treatment-naive CHC patients (P = 0.04). The corresponding rates were comparable between group A and group B CHC patients (P = 0.38). The vast majority of non-responders (10/14, 71.43%) had been infected by genotype-1 of HCV. The seroconversion rates were comparable between group 1 and 2 CHC patients at mo 1(20% versus 26.7%, P = 0.67), mo 2 (46.7% vs 60%,P = 0.46) and mo 7 (86.7% versus 93.3%, P = 0.54) of follow-up.CONCLUSION: The immunogenicity of HBV vaccine seems to be lower in CHC patients compared to healthy subjects. SVR

  1. The use of detectors based on ionisation recombination in radiation protection

    International Nuclear Information System (INIS)

    Intitial recombination of ionisation in a gas depends on the ionisation density and hence on the linear energy transfer along the tracks of charged particles. This effect can be used as a basis for instruments that respond to different types of ionising radiation approximately in the way required by the quality factor-linear energy transfer relation recommended by the ICRP for use in radiation protection. Empirical instruments based on ionisation recombination that have been used for radiation protection measurements are reviewed, and relations are derived from recombination theory that show that the response of such detectors can be readily predicted. The usefulness of recombination instruments in radiation protection is discussed and their advantages and limitations assessed. It is shown that their main application will be as reference instruments against which other detectors can be calibrated. As an extension to using recombination detectors as reference instruments, the feasibility of specifying radiation quality in terms of ionisation recombination is investigated. (author)

  2. Enhanced specific immune responses by CpG DNA in mice immunized with recombinant hepatitis B surface antigen and HB vaccine

    Directory of Open Access Journals (Sweden)

    Wang Xingtai

    2011-02-01

    Full Text Available Abstract Background Hepatitis B vaccine adjuvant, alum, is generally used for vaccination although it does not stimulate Th1 immunity and 10% of the population has low or no antibody response. Efforts have been continued to find more efficient vaccine adjuvants for better antibody response as well as stimulation of Th1 immunity. Methods CpG DNA was used as an adjuvant for recombinant HBsAg to immunize 6- to 8-week-old female BALB/c mice with or without alum for different dosages. The production of HBsAb, CD80 and CD86 from dendritic cells, and cytokines IL-10, IL12, etc., were analyzed and compared for the performance of immunization. Results 5-20 μg CpG DNA had the best co-stimulation effect of HBsAb serum conversion for mice vaccinated with recombinant expressed HBsAg. The mice vaccinated with recombinant 20 μg CpG DNA and regular vaccine (containing alum adjuvant had the highest concentration of antibody production. IL-12b, IL-12a and IL10 mRNA reached to the peak level between 3 and 6 hours after the CpG DNA induction in splenocytes. The expression levels of CD80 and CD86 leucocyte surface molecules were increased with 20 μg CpG DNA alone or with 20 μg CpG DNA and 4 μg HBsAg. Conclusions Our results confirmed the adjuvant effect of CpG DNA for HBsAg in the mouse model. The increase of IL10 and IL12 production suggested the involvement of Th1 cell activation. The activation of CD80 and CD86 molecules by CpG-ODN might be part of the mechanism of T/B cells coordination and the enhancement of recombinant HBsAg induced immune response.

  3. Evaluation of Serum Anti-Hbs Concentration in Children Vaccinated with Recombinant Hepatitis B Vaccine at Birth

    OpenAIRE

    M Nejad-Ghaderi; Mozafari, A.; J Montazerifar; GH Hassanshahi; HR Rashidi-Nejad; A Jafarzadeh

    2006-01-01

    Introduction: Vaccination with the major surface antigen of hepatitis B virus (HBsAg) induces anti-HBs antibody production and level of 10 IU/L is considered protective. It has been shown that the level of anti-HBs antibody does wane after vaccination. The aim of this study was to evaluate the persistence of anti-HBs antibodies in healthy Iranian children 10 years after primary vaccination. Methods: Blood samples were collected from 146 children, 10 years after completion of primary hepatitis...

  4. Robust manufacturing and comprehensive characterization of recombinant hepatitis E virus-like particles in Hecolin(®).

    Science.gov (United States)

    Zhang, Xiao; Wei, Minxi; Pan, Huirong; Lin, Zhijie; Wang, Kaihang; Weng, Zusen; Zhu, Yibin; Xin, Lu; Zhang, Jun; Li, Shaowei; Xia, Ningshao; Zhao, Qinjian

    2014-07-01

    The hepatitis E virus (HEV) vaccine, Hecolin(®), was licensed in China for the prevention of HEV infection and HEV-related diseases with demonstrated safety and efficacy [1,2]. The vaccine is composed of a truncated HEV capsid protein, p239, as the sole antigen encoded by open reading frame 2 and produced using Escherichia coli platform. The production of this virus-like particle (VLP) form of the antigen was successfully scaled up 50-fold from a bench scale to a manufacturing scale. Product consistency was demonstrated using a combination of biophysical, biochemical and immunochemical methods, which revealed comparable antigen characteristics among different batches. Particle size of the nanometer scale particulate antigen and presence of key epitopes on the particle surface are two prerequisites for an efficacious VLP-based vaccine. The particle size was monitored by several different methods, which showed diameters between 20 and 30nm for the p239 particles. The thermal stability and aggregation propensity of the antigen were assessed using differential scanning calorimetry and cloud point assay under heat stress conditions. Key epitopes on the particulate antigen were analyzed using a panel of murine anti-HEV monoclonal antibodies (mAbs). The immuno reactivity to the mAbs among the different antigen lots was highly consistent when analyzed quantitatively using a surface plasmon resonance technique. Using a sandwich ELISA to probe the integrity of two different epitopes in the antigen, the specific antigenicity of multiple batches was assessed to demonstrate consistency in these critical product attributes. Overall, our findings showed that the antigen production process is robust and scalable during the manufacturing of Hecolin(®). PMID:24892250

  5. Recombination spot identification Based on gapped k-mers.

    Science.gov (United States)

    Wang, Rong; Xu, Yong; Liu, Bin

    2016-01-01

    Recombination is crucial for biological evolution, which provides many new combinations of genetic diversity. Accurate identification of recombination spots is useful for DNA function study. To improve the prediction accuracy, researchers have proposed several computational methods for recombination spot identification. The k-mer feature is one of the most useful features for modeling the properties and function of DNA sequences. However, it suffers from the inherent limitation. If the value of word length k is large, the occurrences of k-mers are closed to a binary variable, with a few k-mers present once and most k-mers are absent. This usually causes the sparse problem and reduces the classification accuracy. To solve this problem, we add gaps into k-mer and introduce a new feature called gapped k-mer (GKM) for identification of recombination spots. By using this feature, we present a new predictor called SVM-GKM, which combines the gapped k-mers and Support Vector Machine (SVM) for recombination spot identification. Experimental results on a widely used benchmark dataset show that SVM-GKM outperforms other highly related predictors. Therefore, SVM-GKM would be a powerful predictor for computational genomics. PMID:27030570

  6. The Expression of Sperm Membrane Peptide-Hepatitis B Surface Antigen Fusion Protein with Recombinant Vaccinia Virus

    Institute of Scientific and Technical Information of China (English)

    杨晓鸣; 赵峰; 严缘昌; 李光地; 汪垣

    1998-01-01

    A synthetic oligonucleotide, HSD-2a, encoding a peptide segment of the extracellular domain of a human sperm membrane protein, YWK-Ⅱ, was fused with hepatitis B surface antigen gene (HBs gene). The fused gene was then cloned to pUC18 plasmid.

  7. Evaluation of immunogenicity and safety of Genevac B: A new recombinant hepatitis b vaccine in comparison with Engerix B and Shanvac B in healthy adults

    Directory of Open Access Journals (Sweden)

    Vijayakumar V

    2004-01-01

    Full Text Available PURPOSE: Genevac B, a new indigenous recombinant hepatitis B vaccine was evaluated for its immunogenicity and safety in comparison with Engerix B (Smithkline Beecham Biologicals, Belgium and Shanvac B (Shantha Biotechnics, India in healthy adult volunteers. METHODS: While 240 study subjects were included in the Genevac B group, 80 each were the subjects for Engerix B and Shanvac B. A three dose regimen of 0,1,2 months was adopted with 20 gm dosage uniformly in all the three groups. Vaccinees were assessed during prevaccination, followup and post vaccination periods for clinical, haematological, biochemical and immunological parameters for safety and immunogenicity. RESULTS: Successful follow-up in all parameters for four months could be achieved in 92.5% (222/240 for Genevac B study subjects and the same was 85% (68/80 and 80% (64/80 for Engerix B and Shanvac B respectively. While 100% seroconversion was observed in all the three groups, the rate of seroprotectivity was 99.5% by Genevac B, 98.5% by Engerix B and 98.4% for Shanvac B. However the difference was not statistically significant (p>0.05. The GMT values of anti HBs after one month of completion of the vaccination were 735.50, 718.23 and 662.20 mIU/mL respectively. No systemic reaction was either seen or reported by the volunteers during the vaccination process of Genevac B and other two vaccines. Clinical, haematological and biochemical safety parameters remained within normal limits throughout the study period. CONCLUSION: The study confirms that Genevac B, the new recombinant Hepatitis B vaccine has the acceptable international standards of safety and immunogenicity.

  8. Phylogenetic analysis of complete genome sequences of hepatitis B virus from an Afro-Colombian community: presence of HBV F3/A1 recombinant strain

    Directory of Open Access Journals (Sweden)

    Alvarado-Mora Mónica V

    2012-10-01

    Full Text Available Abstract Background Hepatitis B virus (HBV infection is one of the most prevalent viral infections in humans and represents a serious public health problem. In Colombia, our group reported recently the presence of subgenotypes F3, A2 and genotype G in Bogotá. The aim of this study was to characterize the HBV genotypes circulating in Quibdó, the largest Afro-descendant community in Colombia. Sixty HBsAg-positive samples were studied. A fragment of 1306 bp (S/POL was amplified by nested PCR. Positive samples to S/POL fragment were submitted to PCR amplification of the HBV complete genome. Findings The distribution of HBV genotypes was: A1 (52.17%, E (39.13%, D3 (4.3% and F3/A1 (4.3%. An HBV recombinant strain subgenotype F3/A1 was found for the first time. Conclusions This study is the first analysis of complete HBV genome sequences from Afro-Colombian population. It was found an important presence of HBV/A1 and HBV/E genotypes. A new recombinant strain of HBV genotype F3/A1 was reported in this population. This fact may be correlated with the introduction of these genotypes in the times of slavery.

  9. 接种重组乙型肝炎疫苗低无应答的研究进展%Advances in the research of low-and non-responsiveness after immunization with recombinant hepatitis B vaccine

    Institute of Scientific and Technical Information of China (English)

    陈小英

    2010-01-01

    接种乙型肝炎(乙肝)疫苗是人群预防乙肝的关键措施,但接种乙肝疫苗后,约5%~10%免疫人群呈现抗体低无应答.此文对接种重组乙肝疫苗低无应答的研究进展作一综述.%The key strategy to prevent hepatitis B is immunization with hepatitis B vaccine,but after immunization.about 5%-10% of the vaccinees are low-and non-responders to hepatitis B vaccine.This review summarizes the research progress in low-and non-responsiveness after recombinant hepatitis B vaccination.

  10. Intradermal vaccination of adults with three low doses (2 µg of recombinant hepatitis B vaccine. II. Persistence of immunity and induction of immunologic memory

    Directory of Open Access Journals (Sweden)

    Maria do Carmo M Elisbão

    2003-12-01

    Full Text Available Of the 110 dentists who had presented seroconversion 50 days after the intradermal application of three 2 µg doses of the Belgian recombinant vaccine against hepatitis B (HB, administered eight years before at an interval of one month between the 1st and 2nd doses and of five months between the 2nd and 3rd doses, 51 were included for the assessment of the persistence of immunity. None of the dentists had hepatitis or had received HB vaccine during this period. All subjects were submitted to serological tests for the detection of the following markers of hepatitis B virus (HBV infection: HBsAg, anti-HBc, HBeAg, anti-HBe, and anti-HBs, with no HBsAg, anti-HBc, HBeAg or anti-HBe being detected. A microparticle enzyme immunoassay (MEIA revealed the presence of anti-HBs at protective titers (> 10 mIU/ml in 42 dentists (82.4%, with the anti-HBs titer being higher than 100 mIU/ml in 36 of them (70.6% (good responders, between 10 and 100 mIU/ml in 6 (11.8% (poor responders, and lower than 10 mIU/ml in 9 (17.6% (non-responders. According to clinical data and serological tests, none of the dentists had presented disease or latent HBV infection during the eight years following the first vaccination. A 2 µg booster dose was administered intradermally to eight dentists with anti-HBs titers lower than 10 mIU/ml (non-responders and to six dentists with titers ranging from 10 to 100 mIU/ml (poor responders; the determination of anti-HBs one month later demonstrated the occurrence of seroconversion in the eight non-responders and an increase in anti-HBs titer in the six poor responders. In summary, the present results demonstrated the prolonged persistence of protection against HBV infection and the development of immunologic memory provided by vaccination against HB - with intra-dermal application of three 2 µg doses of the Belgian recombinant vaccine at 0, 1, and 6 months - carried out eight years before in 51 dentists.

  11. Characterization of an age-response relationship to GSK's recombinant hepatitis B vaccine in healthy adults: An integrated analysis

    OpenAIRE

    Van Der Meeren, Olivier; Crasta, Priya Diana; Cheuvart, Brigitte; De Ridder, Marc

    2015-01-01

    The immune system becomes less effective with age, and older age is associated with an increased susceptibility to diseases and reduced responses to vaccination. Furthermore, some adult populations, such as those with diabetes mellitus, are at increased risk of acute hepatitis B virus (HBV) infection. Decreasing responses to vaccination with advanced age have been described, but it is not known at what age immunogenicity starts to reduce, or until what age immunogenicity remains acceptable (f...

  12. Effects of perimeter recombination on GaAs-based solar cells

    Science.gov (United States)

    Stellwag, T. B.; Dodd, P. E.; Carpenter, M. S.; Lundstrom, M. S.; Pierret, R. F.

    Perimeter recombination currents have been experimentally characterized on GaAs p/n heteroface diodes and solar cells with areas ranging from 2.5 x 10 to the -5th to 0.25 sq cm. Under 1-sun operation at the maximum power point, measurements show that the n = roughly 2 perimeter recombination current component degrades the cell's fill factor but does not greatly affect the open-circuit voltage. The n = roughly 2 perimeter recombination currents are examined theoretically on small-area cells using a two-dimensional drift-diffusion device simulator, PUPHS2D. This model verifies the importance and origin of perimeter recombination in heteroface GaAs-based solar cells. Two methods of reducing the n = roughly 2 perimeter recombination are explored.

  13. The Cascade of Care for an Australian Community-Based Hepatitis C Treatment Service.

    Directory of Open Access Journals (Sweden)

    Amanda J Wade

    Full Text Available Hepatitis C treatment uptake in Australia is low. To increase access to hepatitis C virus treatment for people who inject drugs, we developed a community-based, nurse-led service that linked a viral hepatitis service in a tertiary hospital to primary care clinics, and resulted in hepatitis C treatment provision in the community.A retrospective cohort study of patients referred to the community hepatitis service was undertaken to determine the cascade of care. Logistic regression analyses were used to identify predictors of hepatitis C treatment uptake.Four hundred and sixty-two patients were referred to the community hepatitis service; 344 attended. Among the 279 attendees with confirmed chronic hepatitis C, 257 (99% reported ever injecting drugs, and 124 (48% injected in the last month. Of 201 (72% patients who had their fibrosis staged, 63 (31% had F3-F4 fibrosis. Fifty-five patients commenced hepatitis C treatment; 26 (47% were current injectors and 25 (45% had F3-F4 fibrosis. Nineteen of the 27 (70% genotype 1 patients and 14 of the 26 (54% genotype 3 patients eligible for assessment achieved a sustained virologic response. Advanced fibrosis was a significant predictor of treatment uptake in adjusted analysis (AOR 2.56, CI 1.30-5.00, p = 0.006.Our community hepatitis service produced relatively high rates of fibrosis assessment, hepatitis C treatment uptake and cure, among people who inject drugs. These findings highlight the potential benefits of providing community-based hepatitis C care to people who inject drugs in Australia-benefits that should be realised as direct-acting antiviral agents become available.

  14. A Genetic Map of DICTYOSTELIUM DISCOIDEUM Based on Mitotic Recombination

    OpenAIRE

    Welker, Dennis L.; Williams, Keith L.

    1982-01-01

    A genetic map of the cellular slime mold Dictyostelium discoideum is presented in which 42 loci are ordered on five of the seven linkage groups. Although most of the loci were ordered using standing mitotic crossing-over techniques in which recessive selective markers were employed, use was also made of unselected recombined haploid strains. Consistent with cytological studies in which the chromosomes appear to be acrocentric, only a single arm has been found for each of the five linkage grou...

  15. Mechanism-based bioanalysis and biomarkers for hepatic chemical stress.

    Science.gov (United States)

    Antoine, D J; Mercer, A E; Williams, D P; Park, B K

    2009-08-01

    Adverse drug reactions, in particular drug-induced hepatotoxicity, represent a major challenge for clinicians and an impediment to safe drug development. Novel blood or urinary biomarkers of chemically-induced hepatic stress also hold great potential to provide information about pathways leading to cell death within tissues. The earlier pre-clinical identification of potential hepatotoxins and non-invasive diagnosis of susceptible patients, prior to overt liver disease is an important goal. Moreover, the identification, validation and qualification of biomarkers that have in vitro, in vivo and clinical transferability can assist bridging studies and accelerate the pace of drug development. Drug-induced chemical stress is a multi-factorial process, the kinetics of the interaction between the hepatotoxin and the cellular macromolecules are crucially important as different biomarkers will appear over time. The sensitivity of the bioanalytical techniques used to detect biological and chemical biomarkers underpins the usefulness of the marker in question. An integrated analysis of the biochemical, molecular and cellular events provides an understanding of biological (host) factors which ultimately determine the balance between xenobiotic detoxification, adaptation and liver injury. The aim of this review is to summarise the potential of novel mechanism-based biomarkers of hepatic stress which provide information to connect the intracellular events (drug metabolism, organelle, cell and whole organ) ultimately leading to tissue damage (apoptosis, necrosis and inflammation). These biomarkers can provide both the means to inform the pharmacologist and chemist with respect to safe drug design, and provide clinicians with valuable tools for patient monitoring. PMID:19621999

  16. Simple high-cell density fed-batch technique for high-level recombinant protein production with Pichia pastoris: Application to intracellular production of Hepatitis B surface antigen

    OpenAIRE

    Ross Anton; Lünsdorf Heinrich; Gäbel Thomas; Adnan Ahmad; Gurramkonda Chandrasekhar; Nemani Satish; Swaminathan Sathyamangalam; Khanna Navin; Rinas Ursula

    2009-01-01

    Abstract Background Hepatitis B is a serious global public health concern. Though a safe and efficacious recombinant vaccine is available, its use in several resource-poor countries is limited by cost. We have investigated the production of Hepatitis B virus surface antigen (HBsAg) using the yeast Pichia pastoris GS115 by inserting the HBsAg gene into the alcohol oxidase 1 locus. Results Large-scale production was optimized by developing a simple fed-batch process leading to enhanced product ...

  17. Protection against hepatitis B by the Butang® recombinant vaccine in newborn children in South Brazil

    Directory of Open Access Journals (Sweden)

    Aline Paula Isolani

    2006-08-01

    Full Text Available The prevention of hepatitis B by vaccination is one of the most efficient tools to avoid the transmission of the virus. This study evaluated the immunogenicity of the national vaccine Butang® in children born in Campo Mourão City, state of Paraná, Brazil, aged 7 to 12 months, by determining the anti-HBsAg antibodies levels after completion of the National Immunization Program Protocol for hepatitis B. All 70 children evaluated by the MEIA method (immune-enzymatic micro particles showed seroconversion to the Butang® vaccine. Nine children (12.9% presented a low response, with anti-HBs titers between 11 and 100 mUI/ml; 39 children (55.7% showed a good response to the vaccine, with titers between 101 and 1000 mUI/ml; and 22 children (31.4% showed antibodies titers higher than 1000 mUI/ml. The mean titer of the anti-HBs antibody titers was 1408.1 ± 2870.26 mUI/ml (15.7 to 19560.0 mUI/ml. The levels of antibodies produced by the prematurely-born children were not statistically different from those found in the newborns. Fifty-five children were also evaluated through the ELFA method (ELISA with a final detection in fluorescence, which presented similar results. The results obtained in our study corroborated the effectiveness of the national vaccine Butang® in newborn children of Campo Mourão City, Paraná, even if they were premature.

  18. Interaction of hepatic microsomal epoxide hydrolase derived from a recombinant baculovirus expression system with an azarene oxide and an aziridine substrate analogue.

    Science.gov (United States)

    Lacourciere, G M; Vakharia, V N; Tan, C P; Morris, D I; Edwards, G H; Moos, M; Armstrong, R N

    1993-03-16

    A recombinant baculovirus (vEHX) encoding rat hepatic microsomal epoxide hydrolase has been constructed. Infection of Spodoptera frugiperda (Sf9) cells with the recombinant virus results in the expression of the enzyme at a level estimated to be between 5% and 10% of the cellular protein. The enzyme, which can be purified in 15% yield by a simple three-step procedure involving detergent extraction, DEAE-cellulose chromatography, and removal of the detergent on hydroxylapatite, has physical and kinetic properties very close to those of the enzyme obtained from rat liver microsomes. The interaction of the enzyme with two nitrogen-containing analogues of the substrate phenanthrene 9,10-oxide (1) was investigated in order to delineate the contributions of the oxirane group and the hydrophobic surface of the substrate to substrate recognition. The enzyme exhibits altered kinetic properties toward 1,10-phenanthroline 5,6-oxide (2) in which the biphenyl group of 1 is replaced with a bipyridyl group, suggesting that hydrophobic interaction between the complementary surfaces of the substrate and active site has an influence on catalysis. The conjugate acid of the aziridine analogue of 1, phenanthrene 9,10-imine (3), in which the oxirane oxygen is replaced with NH, has a pKa of 6.1, which allows the characterization of both the neutral and protonated aziridine (3H+) as substrate analogues for the enzyme. The pH dependence of the solvolysis reveals that 3H+ rearranges to a 65/35 mixture of 9-aminophenanthrene and 9-amino-10-hydroxy-9,10-dihydrophenanthrene 10(3)-fold faster than does 3. The neutral aziridine is a competitive inhibitor (Ki = 26 microM) of the enzyme at pH 8.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8383521

  19. Survey for immunization effectiveness of 60μg recombinant hepatitis B vaccine(yeast) in nature adults%成人60μg重组乙肝疫苗免疫效果观察

    Institute of Scientific and Technical Information of China (English)

    高雪峰; 闫长伟

    2014-01-01

    目的:探讨成人接种60μg重组乙型肝炎疫苗(酿酒酵母)的免疫成功率和效果。方法选择我县200名16岁以上已进行乙肝疫苗免疫且没有抗体产生的人群,由深圳康泰生物制品股份有限公司提供60μg重组乙型肝炎疫苗220支,批号20100732-1,一针接种后1个月采血,进行血清免疫学检测。结果接种人群均未出现异常反应。接种一针后1个月,阳转率达98%,抗-HBs几何平均滴度(GMT)为150.88 mIU/mL。结论60μg重组乙型肝炎疫苗安全性良好,免疫成功率高,可用于成人乙肝疫苗加强免疫。%Objective The immunization success rate and effectiveness for 60μg recombinant hepatitis B vaccine (yeast) among adults are researched for. Methods 200 healthy adults aged above 16 years in Baishui county and vaccinated hepatitis B vaccines for normal dose but without hepatitis B surface antibody were sampled , vaccinated with one dose 60μg recombinant hepatitis B (HepB) vaccine (yeast), batch 20100732-1, which is produced by BioKangtai, Shenzhen,Guangdong Province, one month after serum immunological observation was conducted. Results no local reaction and systemic reaction happened among adults were observed. One month after inoculating one dose of 60μg recombinant hepatitis B (HepB) vaccine (yeast), the seroconversion rate of resistant hepatitis B surface antigen (anti-HBsAg antibody) reached 98%, the geometric mean titers(GMTs) of anti-HBsAg antibody were 150.88mIU/mL. Conclusions The 60μg recombinant HepB vaccine (yeast) has good safety, high immunization success rate, and enable to be applied for adult hepatitis B vaccine immunization.

  20. Inhibitor-Based Therapeutics for Treatment of Viral Hepatitis

    Science.gov (United States)

    Dey, Debajit; Banerjee, Manidipa

    2016-01-01

    Abstract Viral hepatitis remains a significant worldwide threat, in spite of the availability of several successful therapeutic and vaccination strategies. Complications associated with acute and chronic infections, such as liver failure, cirrhosis and hepatocellular carcinoma, are the cause of considerable morbidity and mortality. Given the significant burden on the healthcare system caused by viral hepatitis, it is essential that novel, more effective therapeutics be developed. The present review attempts to summarize the current treatments against viral hepatitis, and provides an outline for upcoming, promising new therapeutics. Development of novel therapeutics requires an understanding of the viral life cycles and viral effectors in molecular detail. As such, this review also discusses virally-encoded effectors, found to be essential for virus survival and replication in the host milieu, which may be utilized as potential candidates for development of alternative therapies in the future. PMID:27777893

  1. [Study of clinical character and medicinal therapy of viral hepatitis in hospital based on real world].

    Science.gov (United States)

    Li, Yun-ru; Wang, Lian-xin; Xie, Yan-ming; Yang, Wei; Wang, Zhuo-yue; Yi, Dan-hui; Wang, Yong-yan

    2014-09-01

    Viral hepatitis was the most common infectious disease in china. But the diagnosis and treatment were varied because the viral hepatitis patients were hospitalized in different kinds of hospital such as infectious disease hospital, general hospital and Chinese medical hospital. It was necessary to know clinical characters and information of viral hepatitis patients in different hospitals. The general information, subtype distribution, prognosis, complication, medication and relations of onset with solar term from 41 180 viral hepatitis patients based on HIS data were analyzed. It was found that the age of patients between 18 to 59 years old was most; most patients were males. The national basic medical insurance was the most type of payment. The outcome of viral hepatitis in the youth and female were better than that in the old and male. Acute hepatitis was easer to restore than chronic hepatitis. Liver cirrhosis and hepatocellular carcinoma were the two most complications. The peak of onset was during summer solstice, slight heat and great heat. The most common Chinese medicine was Diammonium glycyrrhizinate and the most common western medicine was reduced glutathione. The combination of D. glycyrrhizinate with reduced glutathione, polyene phosphatidylcholine and thymosin was the main pattern. But It was not knew if the combination of western and Chinese medicine was the most effective therapy to protect liver function. It was necessary to take deeply research of the relationship between the combination therapy and their effectiveness.

  2. Evaluation of enzyme immunoassay for hepatitis B virus DNA based on anti-double-stranded DNA.

    OpenAIRE

    F. Garcia(Helsinki U); Bernal, M.C.; Leyva, A.; Piedrola, G.; Maroto, M C

    1995-01-01

    We have evaluated a new enzyme immunoassay technology to detect the products of PCR-based amplification that may be applicable to routine testing of hepatitis B virus (HBV) DNA. Two hundred eight serum samples were studied: 73 were basal samples and 135 were sequential serum samples from patients with chronic hepatitis, some of whom were being treated with alpha interferon. We compared the new detection method (PCR-DNA enzyme immunoassay [DEIA]) with dot blot hybridization performed without p...

  3. Immunogenicity of three recombinant hepatitis B vaccines administered to students in three doses containing half the antigen amount routinely used for adult vaccination

    Directory of Open Access Journals (Sweden)

    Baldy José Luís da Silveira

    2004-01-01

    Full Text Available We evaluated the immunogenicity of three recombinant hepatitis B vaccines, one Brazilian (Butang, Instituto Butantan and two Korean vaccines (Euvax-B, LG Chemical Ltd. and Hepavax-Gene, Greencross Vaccine Corp., administered intramuscularly to students aged 17 to 19 years in three 10-µg doses (corresponding to half the amount of antigen routinely used for adult vaccination at intervals of one month between the first and second dose, and of four months between the second and third dose. A total of 316 students non-reactive for any serological marker of hepatitis B virus infection were vaccinated: 77 (24.4% with the Butang vaccine, 71 (22.5% with Euvax-B, 85 (26.9% with Hepavax-Gene and, for comparison, 83 (26.2% with Engerix-B (GlaxoSmithKline, whose efficacy in young adults at the dose used here has been confirmed in previous studies. Similar seroconversion rates (anti-HBs > 10 mIU/mL about one month after application of the third dose were obtained for the Butang, Euvax-B, Hepavax-Gene and Engerix-B vaccines (96.2%, 98.6%, 96.5% and 97.6%, respectively. The frequency of good responders (anti-HBs > 100 mIU/mL was also similar among students receiving the four vaccines (85.8%, 91.6%, 89.4% and 89.2%, respectively. The geometric mean titers (GMT of anti-HBs about one month after the third dose obtained with these vaccines were 727.78 ± 6.46 mIU/mL, 2009.09 ± 7.16 mIU/mL, 1729.82 ± 8.85 mIU/mL and 2070.14 ± 11.69 mIU/mL, respectively. The GMT of anti-HBs induced by the Euvax-B and Engerix-B vaccines were higher than those obtained with the Butang vaccine (p < 0.05; this difference was not significant when comparing the other vaccines two-by-two. No spontaneous adverse effects attributable to the application of any dose of the four vaccines were reported.

  4. Intradermal vaccination of adults with three low doses (2 µg of recombinant hepatitis B vaccine. I. Seroconversion rate and adverse effects

    Directory of Open Access Journals (Sweden)

    Baldy José Luís da S

    2003-01-01

    Full Text Available A total of 250 dentists (53.6% men and 46.4% women, with a mean age of 35.1 ± 9.8 years, were submitted to serological tests for the diagnosis of hepatitis B (HB - HBsAg, anti-HBs, anti-HBc, HBeAg, and anti-HBe - using a radioimmunoassay. One or more of these markers were detected in 78 individuals (31.2% who were excluded from the group to be vaccinated. Of the 172 HB-susceptible individuals, 135 (78.5% responded to the call and were intradermally injected with three 2 µg doses of the Belgian HB recombinant vaccine, applied at an interval of one month between the 1st and 2nd dose and of five months between the 2nd and 3rd dose. A new determination of HB markers carried out 50 days after the 3rd dose showed that 110 (81.5% individuals had become anti-HBs positive (65.5% good responders and 34.5% poor responders. Mean serum anti-HBs titer of these 110 dentists was 42.4 U S/N, similar in both sexes. The adverse effects analyzed in 106 dentists were: (a local: pain (12.3%, burning sensation (14.1%, pruritus (25.5%, erythema (28.3%, local heat (18.9%, and a hypochromic spot (32.1%; (b systemic (4.7%: discomfort in two patients, and fever, anorexia, and asthenia in one patient each. Intradermal administration of a fourth 2 µg vaccine dose to 39 dentists (poor or non-responders increased the total number of anti-HBs-positive individuals from 110 (81.5% to 114 (84.4%, with the number of good responders increasing from 72 (65.5% to 85 (74.6%. We conclude that the Belgian recombinant vaccine applied in the scheme used here induces a high rate of seroconversion and causes only mild and transitory adverse effects.

  5. Recombinant protein- and synthetic peptide-based immunoblot test for diagnosis of neurocysticercosis.

    Science.gov (United States)

    Noh, John; Rodriguez, Silvia; Lee, Yeuk-Mui; Handali, Sukwan; Gonzalez, Armando E; Gilman, Robert H; Tsang, Victor C W; Garcia, Hector H; Wilkins, Patricia P

    2014-05-01

    One of the most well-characterized tests for diagnosing neurocysticercosis (NCC) is the enzyme-linked immunoelectrotransfer blot (EITB) assay developed at the CDC, which uses lentil lectin-bound glycoproteins (LLGP) extracted from Taenia solium cysticerci. Although the test is very reliable, the purification process for the LLGP antigens has been difficult to transfer to other laboratories because of the need for expensive equipment and technical expertise. To develop a simpler assay, we previously purified and cloned the diagnostic glycoproteins in the LLGP fraction. In this study, we evaluated three representative recombinant or synthetic antigens from the LLGP fraction, individually and in different combinations, using an immunoblot assay (recombinant EITB). Using a panel of 249 confirmed NCC-positive and 401 negative blood serum samples, the sensitivity of the recombinant EITB assay was determined to be 99% and the specificity was 99% for diagnosing NCC. We also tested a panel of 239 confirmed NCC-positive serum samples in Lima, Peru, and found similar results. Overall, our data show that the performance characteristics of the recombinant EITB assay are comparable to those of the LLGP-EITB assay. This new recombinant- and synthetic antigen-based assay is sustainable and can be easily transferred to other laboratories in the United States and throughout the world.

  6. Evidence for reduced charge recombination in carbon nanotube/perovskite-based active layers

    Science.gov (United States)

    Bag, Monojit; Renna, Lawrence A.; Jeong, Seung Pyo; Han, Xu; Cutting, Christie L.; Maroudas, Dimitrios; Venkataraman, D.

    2016-10-01

    Using impedance spectroscopy and computation, we show that incorporation of multi-walled carbon nanotubes (MWCNTs) in the bulk of the active layer of perovskite-based solar cells reduces charge recombination and increases the open circuit voltage. An ∼87% reduction in recombination was achieved when MWCNTs were introduced in the planar-heterostructure perovskite solar cell containing mixed counterions. The open circuit voltage (Voc) of perovskite/MWCNTs devices was increased by 70 mV, while the short circuit current density (Jsc) and fill factor (FF) remained unchanged.

  7. [Autoimmune hepatitis].

    Science.gov (United States)

    Ostojić, Rajko

    2003-01-01

    Autoimmune hepatitis is an unresolving, hepatocellular inflammation of unknown cause that is characterized by the presence of periportal hepatitis on histologic examination, tissue autoantibodies in serum, and hypergammaglobulinemia. By international consensus, the designation autoimmune hepatitis has replaced alternative terms for the condition. Three types of autoimmune hepatitis have been proposed based on immunoserologic findings. Type 1 autoimmune hepatitis is characterized by the presence of antinuclear antibodies (ANA) or smooth muscle antibodies (SMA) (or both) in serum. Seventy percent of patients with type 1 of autoimmune hepatitis are women. This type is the most common form and accounts for at least 80% of cases. Type 2 is characterized by the presence of antibodies to liver-kidney microsome type 1 (anti-LKM1) in serum. Patients with this type of autoimmune hepatitis are predominantly children. Type 3 autoimmune hepatitis is characterized by the presence of antibodies to soluble liver antigen (anti-SLA) in serum. There are no individual features that are pathognomonic of autoimmune hepatitis, and its diagnosis requires the confident exclusion of other conditions. The large majority of patients show satisfactory response to corticosteroid (usually prednisone or prednisolone) therapy. For the past 30 years it has been customary to add azathioprine as a "steroid sparing" agent to allow lower doses of steroids to be used and remission, once achieved, can be sustained in many patients with azathioprine alone after steroid withdrawal. Patients with autoimmune hepatitis who have decompensated during or after corticosteroid therapy are candidates for liver transplantation.

  8. High Seroprotection Rate Induced by Intradermal Administration of a Recombinant Hepatitis B Vaccine in Young Healthy Adults: Comparison with Standard Intramuscular Vaccination

    International Nuclear Information System (INIS)

    Intradermal (ID) vaccination has been proposed as a cost-saving alternative for administration of Hepatitis B (HB) vaccine to implement of mass vaccination of high-risk groups, particularly in developing countries. Therefore, the effectiveness of ID vaccination needs to be evaluated and verified in different ethnic backgrounds. The present study is a randomized trail using a recombinant vaccine (Heberbiovac) to compare immunogenecity and safety of an intradermal low-dose (4 μg) with standard dose (20 μg) of intramuscular (IM) vaccination in healthy Iranian population. Participants were 143 healthy Iranian medical and nursing students randomly allocated to ID or IM vaccination group. The vaccine was inoculated at 0, 1 and 6 months intervals. Serum samples were collected 1 month after the last vaccination and the anti-HBs response was determined using ELISA. The overall seroprotection rate (anti-HBs level ≥ 10IU/L) was 97.3% for ID vaccination group, which was not different from that of IM vaccination group (98.55%)(p= 0.99). Similarly, geometric mean titers (GMT) of anti-HBs were not significantly different between ID (1164.1IU/L) and IM (1071.8IU/L) vaccination groups (p= 0.4). There was no significant difference in seroprotection rate and GMT of anti-HBs between sexes. Although induration and hyperpigmentation at the site of injection were more frequently observed in ID vaccination group, no other clinically adverse effects were observed in both vaccination groups. We conclude that the ID route, which would require one-fifth of the standard dose, would be suitable for use in certain groups such as high-risk adults when the cost of vaccine is the inhibiting factor for mass vaccination

  9. Yeast recombination-based cloning as an efficient way of constructing vectors for Zymoseptoria tritici

    OpenAIRE

    Kilaru, S.; Steinberg, G.

    2015-01-01

    Highlights • Yeast recombination-based cloning (YRBC) is a reliable and inexpensive way of generating plasmids. • We provide 4 vectors for YRBC that a cover different resistance genes. • Using this technique promises rapid generation of molecular tools to study Z. tritici.

  10. A micro-scale hot-surface device based on non-radiative carrier recombination

    NARCIS (Netherlands)

    Kovalgin, A.Y.; Holleman, J.; Iordache, G.

    2004-01-01

    This work employs the idea of making micro-scale hot-surface devices (e.g. sensors, flow meters, micro reactors, etc) based on generation of heat due to nonradiative recombination of carriers in a thin (13 nm) poly silicon surface layer. An important part of the device is a nano-scale (10-100 nm) co

  11. Convergence of a Recombination-Based Elitist Evolutionary Algorithm on the Royal Roads Test Function

    CERN Document Server

    Ter-Sarkisov, Aram

    2011-01-01

    We present an analysis of the performance of an elitist Evolutionary algorithm using a recombination operator known as 1-Bit-Swap on the Royal Roads test function based on a population. We derive complete, approximate and asymptotic convergence rates for the algorithm. The complete model shows the benefit of the size of the population and re- combination pool.

  12. Assessment of a Flavone-Polysaccharide Based Prescription for Treating Duck Virus Hepatitis.

    Directory of Open Access Journals (Sweden)

    Hongxu Du

    Full Text Available Because polysaccharide and flavone ingredients display good antiviral activity, we developed a flavone/polysaccharide-containing prescription that would be effective against duck viral hepatitis (DVH and investigated its hepatoprotective effects. Flavones were derived from Hypericum japonicum (HJF (entire herb of Hypericum japonicum Thunb and Salvia plebeia (SPF (entire herb of Salvia plebeia R. Br., and polysaccharides were derived from Radix Rehmanniae Recens (RRRP (dried root of Rehmannia glutinosa Libosch. This prescription combination was based on the theory of syndrome differentiation and treatment in traditional Chinese veterinary medicine. In vitro and in vivo experiments were conducted using the three single ingredients compared to the combined HRS prescription to determine their anti-duck hepatitis A viral (anti-DHAV activity. The results showed that all experimental conditions displayed anti-DHAV activity, but the HRS prescription presented the best effect. To further investigate the hepatoprotective effect of the HRS prescription on DHAV-induced hepatic injury, we tested the mortality rate, the hepatic pathological severity score, plasma biochemical indexes of hepatic function, blood DHAV gene expression levels and peroxidation damage evaluation indexes and then analyzed correlations among these indexes. The results demonstrated that the HRS prescription significantly decreased the mortality rate, reduced the severity of hepatic injury, decreased the hepatic pathological severity score, depressed blood DHAV gene expression levels, and returned the indexes of hepatic function and peroxidation almost to a normal level. These results indicate that the HRS prescription confers an outstanding hepatoprotective effect, and we expect that it will be developed into a new candidate anti-DHAV drug.

  13. ¹³C-based metabolic flux analysis of recombinant Pichia pastoris.

    Science.gov (United States)

    Ferrer, Pau; Albiol, Joan

    2014-01-01

    Overexpression of a foreign protein may negatively affect several cell growth parameters, as well as cause cellular stress. Central (or core) metabolism plays a crucial role since it supplies energy, reduction equivalents, and precursor molecules for the recombinant product, cell's maintenance, and growth needs. However, the number of quantitative physiology studies of the impact of recombinant protein production on the central metabolic pathways of yeast cell factories has been traditionally rather limited, thereby hampering the application of rational strain engineering strategies targeting central metabolism.The development and application of quantitative physiology and modelling tools and methodologies is allowing for a systems-level understanding of the effect of bioprocess parameters such as growth rate, temperature, oxygen availability, and substrate(s) choice on metabolism, and its subsequent interactions with recombinant protein synthesis, folding, and secretion.Here, we review the recent developments and applications of (13)C-based metabolic flux analysis ((13)C-MFA) of Pichia pastoris and the gained understanding of the metabolic behavior of this yeast in recombinant protein production bioprocesses. We also discuss the potential of multilevel studies integrating (13)C-MFA with other omics analyses, as well as future perspectives on the metabolic modelling approaches to study and design metabolic engineering strategies for improved protein production.

  14. Epidemiology of zoonotic hepatitis E: a community-based surveillance study in a rural population in China.

    Directory of Open Access Journals (Sweden)

    Feng-Cai Zhu

    Full Text Available BACKGROUND: Hepatitis E is caused by two viral genotype groups: human types and zoonotic types. Current understanding of the epidemiology of the zoonotic hepatitis E disease is founded largely on hospital-based studies. METHODS: The epidemiology of hepatitis E was investigated in a community-based surveillance study conducted over one year in a rural city in eastern China with a registered population of 400,162. RESULTS: The seroprevalence of hepatitis E in the cohort was 38%. The incidence of hepatitis E was 2.8/10,000 person-years. Totally 93.5% of the infections were attributed to genotype 4 and the rest, to genotype 1. Hepatitis E accounted for 28.4% (102/359 of the acute hepatitis cases and 68.9% (102/148 of the acute viral hepatitis cases in this area of China. The disease occurred sporadically with a higher prevalence during the cold season and in men, with the male-to-female ratio of 3∶1. Additionally, the incidence of hepatitis E increased with age. Hepatitis B virus carriers have an increased risk of contracting hepatitis E than the general population (OR = 2.5, 95%CI 1.5-4.2. Pre-existing immunity to hepatitis E lowered the risk (relative risk  = 0.34, 95% CI 0.21-0.55 and reduced the severity of the disease. CONCLUSIONS: Hepatitis E in the rural population of China is essentially that of a zoonosis due to the genotype 4 virus, the epidemiology of which is similar to that due to the other zoonotic genotype 3 virus.

  15. Pre-clinical evaluation of a novel nanoemulsion-based hepatitis B mucosal vaccine.

    Directory of Open Access Journals (Sweden)

    Paul E Makidon

    Full Text Available BACKGROUND: Hepatitis B virus infection remains an important global health concern despite the availability of safe and effective prophylactic vaccines. Limitations to these vaccines include requirement for refrigeration and three immunizations thereby restricting use in the developing world. A new nasal hepatitis B vaccine composed of recombinant hepatitis B surface antigen (HBsAg in a novel nanoemulsion (NE adjuvant (HBsAg-NE could be effective with fewer administrations. METHODOLOGY AND PRINCIPAL FINDINGS: Physical characterization indicated that HBsAg-NE consists of uniform lipid droplets (349+/-17 nm associated with HBsAg through electrostatic and hydrophobic interactions. Immunogenicity of HBsAg-NE vaccine was evaluated in mice, rats and guinea pigs. Animals immunized intranasally developed robust and sustained systemic IgG, mucosal IgA and strong antigen-specific cellular immune responses. Serum IgG reached > or = 10(6 titers and was comparable to intramuscular vaccination with alum-adjuvanted vaccine (HBsAg-Alu. Normalization showed that HBsAg-NE vaccination correlates with a protective immunity equivalent or greater than 1000 IU/ml. Th1 polarized immune response was indicated by IFN-gamma and TNF-alpha cytokine production and elevated levels of IgG(2 subclass of HBsAg-specific antibodies. The vaccine retains full immunogenicity for a year at 4 degrees C, 6 months at 25 degrees C and 6 weeks at 40 degrees C. Comprehensive pre-clinical toxicology evaluation demonstrated that HBsAg-NE vaccine is safe and well tolerated in multiple animal models. CONCLUSIONS: Our results suggest that needle-free nasal immunization with HBsAg-NE could be a safe and effective hepatitis B vaccine, or provide an alternative booster administration for the parenteral hepatitis B vaccines. This vaccine induces a Th1 associated cellular immunity and also may provide therapeutic benefit to patients with chronic hepatitis B infection who lack cellular immune

  16. Prevalence of non-responsiveness to an indigenous recombinant hepatitis B vaccine: A study among South Indian health care workers in a tertiary hospital

    Directory of Open Access Journals (Sweden)

    R J Thomas

    2015-01-01

    Full Text Available Background and Aim: Health care workers (HCW are at higher risk of contracting HBV infection. Non-response to HBV vaccine is one of the major impediments to prevent healthcare associated HBV infection (HAHI. We estimated the prevalence of non-responsiveness to initial 3-dose regimen of an indigenous recombinant HBV vaccine (GeneVac-B among South Indian HCWs and typed the HLA in non-responders. Study Design and Method: Of the 778 subjects screened over 1 year, 454 completed all three doses of the hepatitis B vaccination. Anti-HBs titers were estimated by microparticle enzyme immunoassay AxSYM AUSAB, (Abbott, Germany. HLA typing was done using SSP-PCR assay AllSet+™ Gold SSP (Invitrogen, USA. Results: The overall seroconversion rate (anti-HBs > 10 mIU/mL was 98.89% wherein 90.8% had titers >1000mIU/mL, 7.6% had titers 100-1000mIU/mL, 0.43% had titers < 100 mIU/mL and 1.1% were non-responsive (<10 mIU/mL to the initial 3-dose regimen. Antibody titers <1000 mIU/mL were significantly associated with the highest quartile of body mass index (BMI (P < 0.001. We found no significant difference in seroprotection rate between gender (P = 0.088. There was no difference in seroprotection rates among various ethnic groups (P = 0.62. Subjects who were non-responsive in our study had at least one HLA allele earlier known to be associated with non-responsiveness to the vaccine. Conclusion: Our findings suggest that non-response to HBV vaccine is not a major impediment to prevent HAHI. Robust seroprotection rates can be achieved using this indigenous HBV vaccine. However, gender and BMI might influence the level of anti-HBs titers. We recommend the use of this cost effective HBV vaccine as well as postvaccination anti-HBs testing to prevent HAHI among HCWs.

  17. Non-negative constraint for image-based breathing gating in ultrasound hepatic perfusion data

    Science.gov (United States)

    Wu, Kaizhi; Ding, Mingyue; Chen, Xi; Deng, Wenjie; Zhang, Zhijun

    2015-12-01

    Images acquired during free breathing using contrast enhanced ultrasound hepatic perfusion imaging exhibits a periodic motion pattern. It needs to be compensated for if a further accurate quantification of the hepatic perfusion analysis is to be executed. To reduce the impact of respiratory motion, image-based breathing gating algorithm was used to compensate the respiratory motion in contrast enhanced ultrasound. The algorithm contains three steps of which respiratory kinetics extracted, image subsequences determined and image subsequences registered. The basic performance of the algorithm was to extract the respiratory kinetics of the ultrasound hepatic perfusion image sequences accurately. In this paper, we treated the kinetics extracted model as a non-negative matrix factorization (NMF) problem. We extracted the respiratory kinetics of the ultrasound hepatic perfusion image sequences by non-negative matrix factorization (NMF). The technique involves using the NMF objective function to accurately extract respiratory kinetics. It was tested on simulative phantom and used to analyze 6 liver CEUS hepatic perfusion image sequences. The experimental results show the effectiveness of our proposed method in quantitative and qualitative.

  18. GENERALIZED REGRESSION NEURAL NETWORK BASED EXPERT SYSTEM FOR HEPATITIS B DIAGNOSIS

    Directory of Open Access Journals (Sweden)

    C. Mahesh

    2014-01-01

    Full Text Available Hepatitis B is a potentially life-threatening liver infection caused by the hepatitis B virus. The virus interferes with the function of the liver while replicating in hepatocytes. It is a major global health problem and the most serious type of viral hepatitis. Chronic liver disease is caused by viral hepatitis and putting people at high risk of death from cirrhosis of the liver and liver cancer. Medical information available is extensive and which is utilized by the clinical specialists. The ranging of information is from details of clinical symptoms to various types of biochemical data. Information provided by each data is evaluated and assigned to a particular pathology during the diagnostic process. Artificial intelligence methods especially computer aided diagnosis and artificial neural networks can be employed to streamline the diagnostic process. These adaptive learning algorithms can handle diverse types of medical data and integrate them into categorized outputs. Artificial neural networks are finding many uses in the medical diagnosis application. In this study we have proposed a Generalized Regression Neural Network (GRNN based expert system for the diagnosis of the hepatitis B virus disease. The system classifies each patient into infected and non-infected. If infected then how severe it is in terms of intensity rate.

  19. CT-based temperature monitoring during hepatic RF ablation : Feasibility in an animal model

    NARCIS (Netherlands)

    Bruners, Philipp; Pandeya, Ganga D.; Levit, Elena; Roesch, Eva; Penzkofer, Tobias; Isfort, Peter; Schmidt, Bernhardt; Greuter, Marcel J. W.; Oudkerk, Matthijs; Schmitz-Rode, Thomas; Kuhl, Christiane K.; Mahnken, Andreas H.

    2012-01-01

    Purpose: The aim of this paper was to establish non-invasive CT-based temperature monitoring during hepatic radiofrequency (RF) ablation in an ex vivo porcine model followed by transfer of the technique into a feasibility in vivo experiment. Materials and methods: Bipolar RF ablations were performed

  20. Capacity Enhancement of Hepatitis C Virus Treatment through Integrated, Community-Based Care

    Directory of Open Access Journals (Sweden)

    Warren D Hill

    2008-01-01

    Full Text Available BACKGROUND: An estimated 250,000 Canadians are infected with the hepatitis C virus (HCV. The present study describes a cohort of individuals with HCV referred to community-based, integrated prevention and care projects developed in British Columbia. Treatment outcomes are reported for a subset of individuals undergoing antiviral therapy at four project sites.

  1. Advances in the study of multivalent recombinant DNA vaccines utilizing the hepatitis B virus surface antigen gene%乙肝表面抗原载体多价重组核酸疫苗研究进展

    Institute of Scientific and Technical Information of China (English)

    肖婷; 郭根灵; 辛宪云; 魏庆宽

    2012-01-01

    研究表明,乙肝病毒的包膜蛋白HBsAg不仅可以作为疫苗的理想候选分子,还可作为基因工程疫苗的理想载体,用来成功构建多种重组核酸疫苗.本文概述了以乙肝表面抗原为载体,重组或联合其他病毒、寄生虫、细胞因子等其他基因制作多价核酸疫苗的研究进展.%Studies have shown that hepatitis B virus envelope protein HBsAg can be used as an ideal candidate molecule for vaccines and also as an ideal vehicle for genetically engineered vaccines to successfully build a variety of recombinant DNA vaccines. This article provides an overview of advances in recombinant DNA vaccines prepared by using hepatitis B virus surface antigen as a carrier to restructure or join it to other viruses, parasites, cell factors, or other genes.

  2. Development of multiple sclerosis after vaccination against hepatitis B: a study based on human leucocyte antigen haplotypes.

    Science.gov (United States)

    Ozakbas, S; Idiman, E; Yulug, B; Pakoz, B; Bahar, H; Gulay, Z

    2006-09-01

    The aetiology of multiple sclerosis (MS) is still not fully understood. Infectious agents are believed to play a role in the development of this multifactorial disease. Cases in which this disease occurs after administration of both plasma-derived and recombinant hepatitis B vaccines have been reported. In this study, we compared a group of 11 MS patients who developed first clinical symptoms after hepatitis B vaccination (group I) with 71 MS patients who were never vaccinated against hepatitis B and were negative for hepatitis B serology (group II), and 20 healthy controls (group III). Mean age was 27.75 years (19-39) in group I, 30.16 years (18-50) in group II, and 34.4 years (18-50) in group III. Mean attack rate after 2 years was 1.5 in group I and 1.63 in group II. Mean Expanded Disability Status Scale score after 2 years was 1.31 in group I and 1.89 in group II. Human leucocyte antigen (HLA) typing and serology for hepatitis B surface antigen were performed in all groups. In groups I and II, HLA-DR2 was more frequent than in normal healthy subjects. This reflects the general role of HLA in the pathogenesis of MS but suggests that antigen presentation by different HLA is not involved in the development of MS after hepatitis B vaccination. Since there was no difference in the clinical features between vaccinated and nonvaccinated MS patients, this study supports recent reports that hepatitis B vaccination is safe in MS patients and that hepatitis B vaccination is not involved in the development of MS. PMID:16948644

  3. 两种重组乙型肝炎疫苗免疫效果比较%Comparison of the immune effect of two kinds of recombinant hepatitis B vaccine

    Institute of Scientific and Technical Information of China (English)

    苏文娟

    2016-01-01

    To analyze the differences in clinical efficacy and safety of recombinant hepatitis B vaccine saccharomyces cereG visiae and recombinant hepatitis B vaccine hansenula polymorpha.Methods:120 cases of people receiving hepatitis B vaccination from November 2012 to December 2014 were selected as research subjects and randomly divided into group A 60 cases and group B 60 cases according to different vaccine they received.Group A received hepatitis B vaccine hansenula polymorpha and group B received hepatitis B vaccine saccharomyces cerevisiae.Differences in titer,adverse reactions and so on of two groups after vaccination were compared. Results:Positive conversion,protection level ratio and GMT value of group A after hepatitis B vaccination were higher than those of control group (P<0.05);the probability of local redness and swelling,induration,general fever,tireness,anorexia and so on of group A after hepatitis B vaccination were lower than those of control group (P<0.05).Conclusion:Vaccination of hepatitis B vaccine hanG senula polymorpha for healthy people can obtain betterimmune effect,it is a more ideal hepatitis B vaccine.%目的::分析重组酿酒酵母乙肝疫苗与重组汉逊酵母乙肝疫苗的临床应用效果.方法选取2012年11月-2014年12月间接受乙肝疫苗接种的人群120例,根据接受疫苗不同分为 A 组及 B 组各60例.A 组接受汉逊酵母乙型肝炎疫苗、B 组患者接受酿酒酵母乙型肝炎疫苗.对比两组患者的疫苗接种后滴度、不良反应等情况.结果:A 组患者接受乙肝疫苗接种后的阳转、保护水平比例、GMT 值均高于对照组患者(P<0.05);A 组患者接受乙肝疫苗后的局部红肿硬结,全身发热、倦怠、厌食等概率均低于 B 组患者(P<0.05).结论:汉逊酵母乙型肝炎疫苗用于健康人群接种后可以获得更好的免疫效果,是更为理想的乙型肝炎疫苗.

  4. Comparison of the immune effect of two kinds of recombinant hepatitis B vaccine%两种重组乙型肝炎疫苗免疫效果比较

    Institute of Scientific and Technical Information of China (English)

    苏文娟

    2016-01-01

    目的::分析重组酿酒酵母乙肝疫苗与重组汉逊酵母乙肝疫苗的临床应用效果.方法选取2012年11月-2014年12月间接受乙肝疫苗接种的人群120例,根据接受疫苗不同分为 A 组及 B 组各60例.A 组接受汉逊酵母乙型肝炎疫苗、B 组患者接受酿酒酵母乙型肝炎疫苗.对比两组患者的疫苗接种后滴度、不良反应等情况.结果:A 组患者接受乙肝疫苗接种后的阳转、保护水平比例、GMT 值均高于对照组患者(P<0.05);A 组患者接受乙肝疫苗后的局部红肿硬结,全身发热、倦怠、厌食等概率均低于 B 组患者(P<0.05).结论:汉逊酵母乙型肝炎疫苗用于健康人群接种后可以获得更好的免疫效果,是更为理想的乙型肝炎疫苗.%To analyze the differences in clinical efficacy and safety of recombinant hepatitis B vaccine saccharomyces cereG visiae and recombinant hepatitis B vaccine hansenula polymorpha.Methods:120 cases of people receiving hepatitis B vaccination from November 2012 to December 2014 were selected as research subjects and randomly divided into group A 60 cases and group B 60 cases according to different vaccine they received.Group A received hepatitis B vaccine hansenula polymorpha and group B received hepatitis B vaccine saccharomyces cerevisiae.Differences in titer,adverse reactions and so on of two groups after vaccination were compared. Results:Positive conversion,protection level ratio and GMT value of group A after hepatitis B vaccination were higher than those of control group (P<0.05);the probability of local redness and swelling,induration,general fever,tireness,anorexia and so on of group A after hepatitis B vaccination were lower than those of control group (P<0.05).Conclusion:Vaccination of hepatitis B vaccine hanG senula polymorpha for healthy people can obtain betterimmune effect,it is a more ideal hepatitis B vaccine.

  5. Yeast recombination-based cloning as an efficient way of constructing vectors for Zymoseptoria tritici.

    Science.gov (United States)

    Kilaru, S; Steinberg, G

    2015-06-01

    Many pathogenic fungi are genetically tractable. Analysis of their cellular organization and invasion mechanisms underpinning virulence determinants profits from exploiting such molecular tools as fluorescent fusion proteins or conditional mutant protein alleles. Generation of these tools requires efficient cloning methods, as vector construction is often a rate-limiting step. Here, we introduce an efficient yeast recombination-based cloning (YRBC) method to construct vectors for the fungus Zymoseptoria tritici. This method is of low cost and avoids dependency on the availability of restriction enzyme sites in the DNA sequence, as needed in more conventional restriction/ligation-based cloning procedures. Furthermore, YRBC avoids modification of the DNA of interest, indeed this potential risk limits the use of site-specific recombination systems, such as Gateway cloning. Instead, in YRBC, multiple DNA fragments, with 30bp overlap sequences, are transformed into Saccharomyces cerevisiae, whereupon homologous recombination generates the vector in a single step. Here, we provide a detailed experimental protocol and four vectors, each encoding a different dominant selectable marker cassette, that enable YRBC of constructs to be used in the wheat pathogen Z. tritici. PMID:26092792

  6. ALKBH1 is dispensable for abasic site cleavage during base excision repair and class switch recombination.

    Science.gov (United States)

    Müller, Tina A; Yu, Kefei; Hausinger, Robert P; Meek, Katheryn

    2013-01-01

    Potential roles of the abasic site lyase activity associated with AlkB homolog 1 (ALKBH1) were assessed by studies focusing on the two cellular processes that create abasic sites as intermediates: base excision repair and class switch recombination. Alkbh1(-/-) pups (lacking exon 3) were born at a lower than expected frequency from heterozygous parents, suggesting a reduced survival rate and non-Mendelian inheritance, and they exhibited a gender bias in favor of males (70% males and 30% females). To study ALKBH1's potential involvement in DNA repair, fibroblasts were isolated from Alkbh1(-/-) mice, spontaneously immortalized and tested for resistance to DNA damaging agents. Alkbh1(-/-) and isogenic cells expressing hALKBH1 showed no difference in survival to the DNA damaging agents methyl-methionine sulfate or H2O2. This result indicates that ALKBH1 does not play a major role in the base excision repair pathway. To assess ALKBH1's role in class switch recombination, splenic B cells were isolated from Alkbh1(-/-) and Alkbh1(+/+) mice and subjected to switching from IgM to IgG1. No differences were found in IgG1 switching, suggesting that Alkbh1 is not involved in class switch recombination of the immunoglobulin heavy chain during B lymphocyte activation. PMID:23825659

  7. ALKBH1 is dispensable for abasic site cleavage during base excision repair and class switch recombination.

    Directory of Open Access Journals (Sweden)

    Tina A Müller

    Full Text Available Potential roles of the abasic site lyase activity associated with AlkB homolog 1 (ALKBH1 were assessed by studies focusing on the two cellular processes that create abasic sites as intermediates: base excision repair and class switch recombination. Alkbh1(-/- pups (lacking exon 3 were born at a lower than expected frequency from heterozygous parents, suggesting a reduced survival rate and non-Mendelian inheritance, and they exhibited a gender bias in favor of males (70% males and 30% females. To study ALKBH1's potential involvement in DNA repair, fibroblasts were isolated from Alkbh1(-/- mice, spontaneously immortalized and tested for resistance to DNA damaging agents. Alkbh1(-/- and isogenic cells expressing hALKBH1 showed no difference in survival to the DNA damaging agents methyl-methionine sulfate or H2O2. This result indicates that ALKBH1 does not play a major role in the base excision repair pathway. To assess ALKBH1's role in class switch recombination, splenic B cells were isolated from Alkbh1(-/- and Alkbh1(+/+ mice and subjected to switching from IgM to IgG1. No differences were found in IgG1 switching, suggesting that Alkbh1 is not involved in class switch recombination of the immunoglobulin heavy chain during B lymphocyte activation.

  8. PCR-based gene synthesis to produce recombinant proteins for crystallization

    Directory of Open Access Journals (Sweden)

    Byrne-Steele Miranda L

    2008-04-01

    Full Text Available Abstract Background Gene synthesis technologies are an important tool for structural biology projects, allowing increased protein expression through codon optimization and facilitating sequence alterations. Existing methods, however, can be complex and not always reproducible, prompting researchers to use commercial suppliers rather than synthesize genes themselves. Results A PCR-based gene synthesis method, referred to as SeqTBIO, is described to efficiently assemble the coding regions of two novel hyperthermophilic proteins, PAZ (Piwi/Argonaute/Zwille domain, a siRNA-binding domain of an Argonaute protein homologue and a deletion mutant of a family A DNA polymerase (PolA. The gene synthesis procedure is based on sequential assembly such that homogeneous DNA products can be obtained after each synthesis step without extensive manipulation or purification requirements. Coupling the gene synthesis procedure to in vivo homologous recombination techniques allows efficient subcloning and site-directed mutagenesis for error correction. The recombinant proteins of PAZ and PolA were subsequently overexpressed in E. coli and used for protein crystallization. Crystals of both proteins were obtained and they were suitable for X-ray analysis. Conclusion We demonstrate, by using PAZ and PolA as examples, the feasibility of integrating the gene synthesis, error correction and subcloning techniques into a non-automated gene to crystal pipeline such that genes can be designed, synthesized and implemented for recombinant expression and protein crystallization.

  9. Ensemble and Arithmetic Recombination-Based Speciation Differential Evolution for Multimodal Optimization.

    Science.gov (United States)

    Hui, Sheldon; Suganthan, Ponnuthurai N

    2016-01-01

    Multimodal optimization problems consists of multiple equal or comparable spatially distributed solutions. Niching and clustering differential evolution (DE) techniques have been demonstrated to be highly effective for solving such problems. The key challenge in the speciation niching technique is to balance between local solution exploitation and global exploration. Our proposal enhances exploration by applying arithmetic recombination with speciation and improves exploitation of individual peaks by applying neighborhood mutation with ensemble strategies. Our novel algorithm, called ensemble and arithmetic recombination-based speciation DE, is shown to either outperform or perform comparably to the state-of-the-art algorithms on 29 common multimodal benchmark problems. Comparable performance is observed only when some problems are solved perfectly by the algorithms in the literature. PMID:25781971

  10. Subunit Protein Vaccine Delivery System for Tuberculosis Based on Hepatitis B Virus Core VLP (HBc-VLP) Particles.

    Science.gov (United States)

    Dhanasooraj, Dhananjayan; Kumar, R Ajay; Mundayoor, Sathish

    2016-01-01

    Despite the development of modern medicine, tuberculosis (TB), caused by the pathogenic bacterium, Mycobacterium tuberculosis (Mtb), remains one of the deadliest diseases. This bacterium can lay dormant in individuals and get activated when immunity goes down and has also shown considerable prowess in mutating into drug resistant forms. The global emergence of such drug resistant Mtb and the lack of efficacy of Bacille Calmette Guérin (BCG), the only vaccine available so far, have resulted in a situation which cries out for a safe and effective tuberculosis vaccine.Number of different strategies has been used for developing new anti-TB vaccines and several protective antigens have been identified so far. One strategy, the use of protein subunits, has the potential to develop into a powerful tuberculosis vaccine, not only because of its efficacy and safety, but also because they are economical. The proper delivery of protein subunit vaccines with adjuvants or novel delivery systems is necessary for inducing protective immune responses. The available adjuvants or delivery systems are inadequate for generating such a response. In the present method, we have constructed a vaccine delivery system for tuberculosis based on Virus-Like Particles (VLPs). Hepatitis B Virus core antigen gene was recombinantly modified using Overlap Extension PCR (OEPCR). The final construct was designed to express HBc-VLP carrying external antigen (fusion VLP). Mycobacterium tuberculosis antigen CFP-10 was used for the construction of fusion VLP. The recombinant gene for the construct was cloned into a pET expression system and transformed into E. coli BL21(DE3) and induced with IPTG to express the protein. The fusion protein was purified using the Histidine tag and allowed to form VLPs. The preformed VLPs were purified by sucrose density gradient centrifugation. The VLPs were characterized using Transmission Electron Microscopy (TEM). PMID:27076312

  11. Induction of anti-human immunodeficiency virus (HIV) neutralizing antibodies in rabbits immunized with recombinant HIV--hepatitis B surface antigen particles.

    OpenAIRE

    Michel, M L; M. Mancini; Sobczak, E.; Favier, V.; Guetard, D; Bahraoui, E M; Tiollais, P

    1988-01-01

    Fragments of the human immunodeficiency virus (HIV) envelope coding region have been fused with the hepatitis B virus envelope middle protein. In this system, HIV antigenic determinants are exposed at the surface of a highly antigenic structure, the hepatitis B surface antigen particle. Immunization of rabbits with these particles elicited antibodies directed against both parts of the hybrid protein. One of the rabbit antisera not only exhibited a neutralizing effect on the original HIV1 isol...

  12. Hepatitis B Vaccination in Bangladesh: a Suggestion Based on Current Evidence

    Directory of Open Access Journals (Sweden)

    Shafquat Mohammed Rafiq

    2006-12-01

    Full Text Available IntroductionThe hepatitis B virus (HBV causes up to a million deaths worldwide and 16 million health care related infections in the tropics each year(1,2, and over 350 million become chronically infected carriers who have no significant liver disease; approximately three quarters of them are in Asia and the western pacific region(3,4. HBV infection is a potentially life threatening condition as many of the affected individuals progress to chronic hepatitis,cirrhosis and hepatocellular carcinoma (HCC(3. In infants and children, acute hepatitis B infection is nearly always asymptomatic, whereas in adults it is usually the opposite. But on the other hand, the risk of becoming chronic carriage is much greater in children than in adults; as many as 90% of infants born to Hepatitis B e Antigen (HBeAg positive mothers become carriers themselves and, therefore, in long term are more likely to developchronic liver disease(5. Currently, though several antiviral drugs are used,there is no reliable curative treatment for HBV once it has been acquired and prevention by universal immunization remains the strategy for reducing the number of acute infections, chronic carriage and the long-term burden from diseases such as HCC(4,6. In 1991, in an attempt to reduce the global impact of HBV infection, WHO recommended that hepatitis B vaccination should be integrated into national immunization programs in all countries(7.Some Asian countries, for instance, Thailand, haveadopted the policy of immunizing children universally against the disease as early as 1992, however many others lagged behind(4.The true prevalence of Hepatitis B in Bangladesh is yet to be ascertained by a reliable study. Data available from different studies show that it ranges between 0.8 and 5.4% depending on the study design, samples and laboratory methods used(8-10.These data were based on detection of HBsAg antigen; the rates would have been higher, had they been based on anti-HBc antibody(11

  13. Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

    Directory of Open Access Journals (Sweden)

    N.S. Sato

    2004-07-01

    Full Text Available Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples, from healthy blood donors (50 samples, individuals with sexually transmitted disease other than syphilis (3 samples, and from individuals with other spirochetal diseases such as Lyme disease (20 samples and leptospirosis (3 samples. The assay had sensitivity of 95.1% (95% CI, 86.1 to 98.7% and a specificity of 94.7% (95% CI, 87.0 to 98.7%; a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

  14. Development of a diphtheria toxin based antiporcine CD3 recombinant immunotoxin.

    Science.gov (United States)

    Wang, Zhirui; Duran-Struuck, Raimon; Crepeau, Rebecca; Matar, Abraham; Hanekamp, Isabel; Srinivasan, Srimathi; Neville, David M; Sachs, David H; Huang, Christene A

    2011-10-19

    Anti-CD3 immunotoxins, which induce profound but transient T-cell depletion in vivo by inhibiting eukaryotic protein synthesis in CD3+ cells, are effective reagents in large animal models of transplantation tolerance and autoimmune disease therapy. A diphtheria toxin based antiporcine CD3 recombinant immunotoxin was constructed by fusing the truncated diphtheria toxin DT390 with two identical tandem single chain variable fragments (scFv) derived from the antiporcine CD3 monoclonal antibody 898H2-6-15. The recombinant immunotoxin was expressed in a diphtheria-toxin resistant yeast Pichia pastoris strain under the control of the alcohol oxidase promoter. The secreted recombinant immunotoxin was purified sequentially with hydrophobic interaction chromatography (Butyl 650 M) followed by strong anion exchange (Poros 50 HQ). The purified antiporcine CD3 immunotoxin was tested in vivo in four animals; peripheral blood CD3+ T-cell numbers were reduced by 80% and lymph node T-cells decreased from 74% CD3+ cells pretreatment to 24% CD3+ cells remaining in the lymph node following 4 days of immunotoxin treatment. No clinical toxicity was observed in any of the experimental swine. We anticipate that this conjugate will provide an important tool for in vivo depletion of T-cells in swine transplantation models. PMID:21866954

  15. Improved serodiagnosis of hepatitis C virus infection with synthetic peptide antigen from capsid protein.

    OpenAIRE

    Hosein, B; Fang, C T; Popovsky, M A; J. Ye; Zhang, M; WANG, C. Y.

    1991-01-01

    Cloning and expression of hepatitis C virus have allowed the development of immunoassays to detect hepatitis C virus infection. However, currently available recombinant fusion protein C100-3 assays, based on a nonstructural protein of the virus, are limited in sensitivity, particularly for detecting acute infection. In this report seroconversion panels showed that an assay based on synthetic peptides, derived from immunodominant regions of both capsid and nonstructural proteins, accelerated h...

  16. Capacity Enhancement of Hepatitis C Virus Treatment through Integrated, Community-Based Care

    OpenAIRE

    Hill, Warren D; Butt, Gail; Alvarez, Maria; Krajden, Mel

    2008-01-01

    BACKGROUND: An estimated 250,000 Canadians are infected with the hepatitis C virus (HCV). The present study describes a cohort of individuals with HCV referred to community-based, integrated prevention and care projects developed in British Columbia. Treatment outcomes are reported for a subset of individuals undergoing antiviral therapy at four project sites.METHODS: Four demonstration projects based on a public health nurse and physician partnership were established in rural and small urban...

  17. Virus-like particle production with yeast: ultrastructural and immunocytochemical insights into Pichia pastoris producing high levels of the Hepatitis B surface antigen.

    OpenAIRE

    Adnan Ahmad; Gurramkonda Chandrasekhar; Lünsdorf Heinrich; Khanna Navin; Rinas Ursula

    2011-01-01

    Abstract Background A protective immune response against Hepatitis B infection can be obtained through the administration of a single viral polypeptide, the Hepatitis B surface antigen (HBsAg). Thus, the Hepatitis B vaccine is generated through the utilization of recombinant DNA technology, preferentially by using yeast-based expression systems. However, the polypeptide needs to assemble into spherical particles, so-called virus-like particles (VLPs), to elicit the required protective immune ...

  18. Serological Assays Based on Recombinant Viral Proteins for the Diagnosis of Arenavirus Hemorrhagic Fevers

    Directory of Open Access Journals (Sweden)

    Masayuki Saijo

    2012-10-01

    Full Text Available The family Arenaviridae, genus Arenavirus, consists of two phylogenetically independent groups: Old World (OW and New World (NW complexes. The Lassa and Lujo viruses in the OW complex and the Guanarito, Junin, Machupo, Sabia, and Chapare viruses in the NW complex cause viral hemorrhagic fever (VHF in humans, leading to serious public health concerns. These viruses are also considered potential bioterrorism agents. Therefore, it is of great importance to detect these pathogens rapidly and specifically in order to minimize the risk and scale of arenavirus outbreaks. However, these arenaviruses are classified as BSL-4 pathogens, thus making it difficult to develop diagnostic techniques for these virus infections in institutes without BSL-4 facilities. To overcome these difficulties, antibody detection systems in the form of an enzyme-linked immunosorbent assay (ELISA and an indirect immunofluorescence assay were developed using recombinant nucleoproteins (rNPs derived from these viruses. Furthermore, several antigen-detection assays were developed. For example, novel monoclonal antibodies (mAbs to the rNPs of Lassa and Junin viruses were generated. Sandwich antigen-capture (Ag-capture ELISAs using these mAbs as capture antibodies were developed and confirmed to be sensitive and specific for detecting the respective arenavirus NPs. These rNP-based assays were proposed to be useful not only for an etiological diagnosis of VHFs, but also for seroepidemiological studies on VHFs. We recently developed arenavirus neutralization assays using vesicular stomatitis virus (VSV-based pseudotypes bearing arenavirus recombinant glycoproteins. The goal of this article is to review the recent advances in developing laboratory diagnostic assays based on recombinant viral proteins for the diagnosis of VHFs and epidemiological studies on the VHFs caused by arenaviruses.

  19. Comparative Study of Monoclonal and Recombinant Antibody-Based Immunoassays for Fungicide Analysis in Fruit juices

    OpenAIRE

    Moreno Tamarit, Mª José; PLANA ANDANI, EMMA; Manclus Ciscar, Juan José; Montoya Baides, Ángel

    2014-01-01

    [EN] A comparative study of the analytical performance of enzyme-linked immunosorbent assays (ELISAs), based on monoclonal and recombinant antibodies, for the determination of fungicide residues in fruit juices has been carried out. To this aim, three murine hybridoma cell lines secreting specific monoclonal antibodies against (RS)-2-(2,4-dichlorophenyl)-3-(1H-1,2,4-triazol-1-yl)propyl-1,1,2,2-tetrafluoroethyl ether (tetraconazole), 2-(4-triazolyl)benzimidazole (thiabendazole), and (RS)-1-(be...

  20. Environment control to improve recombinant protein yields in plants based on Agrobacterium-mediated transient gene expression

    Directory of Open Access Journals (Sweden)

    Naomichi eFujiuchi

    2016-03-01

    Full Text Available Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: 1 recombinant protein content per unit biomass; and 2 recombinant protein productivity per unit area-time at the end of the upstream production. Because environmental factors in the upstream production have impacts on those parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and post-inoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Pre-inoculation environmental factors associated with planting density, light quality and nutrient supply affect plant characteristics such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, post-inoculation environmental factors such as temperature, light intensity and humidity have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the post-inoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain

  1. Highly Efficient JFH1-Based Cell-Culture System for Hepatitis C Virus Genotype 5a: Failure of Homologous Neutralizing-Antibody Treatment to Control Infection

    DEFF Research Database (Denmark)

    Jensen, Tanja B; Gottwein, Judith Margarete; Scheel, Troels Kasper Høyer;

    2008-01-01

    Background. @nbsp; Recently, a hepatitis C virus (HCV) cell-culture system was developed that employed strain JFH1 (genotype 2a), and JFH1-based intra- and intergenotypic recombinants now permit functional studies of the structural genes (Core, E1, and E2), p7, and NS2 of genotypes 1-4. The goal...... was to adapt the system to employ genotype 5. Methods. @nbsp; Huh7.5 cells infected with SA13/JFH1, containing Core-NS2 of strain SA13 (genotype 5a), were monitored for Core expression and for supernatant infectivity and HCV-RNA titers. Adaptive mutations of SA13/JFH1 were identified by sequence analysis...

  2. Recommendations for the Clinical Management of Hepatitis C in Iran: A Consensus-Based National Guideline

    Science.gov (United States)

    Alavian, Seyed Moayed; Hajarizadeh, Behzad; Bagheri Lankarani, Kamran; Sharafi, Heidar; Ebrahimi Daryani, Nasser; Merat, Shahin; Mohraz, Minoo; Mardani, Masoud; Fattahi, Mohamad Reza; Poustchi, Hossein; Nikbin, Mehri; Nabavi, Mahmood; Adibi, Peyman; Ziaee, Masood; Behnava, Bita; Rezaee-Zavareh, Mohammad Saeid; Colombo, Massimo; Massoumi, Hatef; Bizri, Abdul Rahman; Eghtesad, Bijan; Amiri, Majid; Namvar, Ali; Hesamizadeh, Khashayar; Malekzadeh, Reza

    2016-01-01

    Context Hepatitis C virus (HCV) infection is a major public health issue worldwide, including Iran. The new direct-acting antiviral agents (DAAs) with high efficacy have changed the landscape of HCV treatment. This guideline provides updated recommendations for clinical management of HCV infection in Iran. Evidence Acquisition The recommendations of this guideline are based on international and national scientific evidences and consensus-based expert opinion. Scientific evidences were collected through a systematic review of studies that evaluated efficacy and safety of DAA regimens, using PubMed, Scopus and Web of Science. Expert opinion was based on the consensus of Iran Hepatitis Scientific Board (IHSB) in the 3rd national consensus on management of Hepatitis C in Iran, held on 22nd of July 2016. Results Pegylated Interferon alpha (PegIFN), Ribavirin (RBV), Sofosbuvir (SOF), Ledipasvir (LDV) and Daclatasvir (DCV) are currently available in Iran. Pre-treatment assessments include HCV RNA level, HCV genotype and resistance testing, assessment of liver fibrosis, and underlying diseases. In HCV genotype 1 and 4, DCV/SOF and LDV/SOF are recommended. In HCV genotype 2, SOF plus RBV and in HCV genotype 3, DCV/SOF is recommended. Additional care for underlying diseases should be considered. Conclusions Affordable new HCV treatment regimens are available in Iran, providing an opportunity for HCV elimination. Recommendations provided in this current national guideline can facilitate evidence-based management of HCV infection. PMID:27799966

  3. Recommendations for the Clinical Management of Hepatitis C in Iran: A Consensus-Based National Guideline

    Directory of Open Access Journals (Sweden)

    Alavian

    2016-08-01

    Full Text Available Context Hepatitis C virus (HCV infection is a major public health issue worldwide, including Iran. The new direct-acting antiviral agents (DAAs with high efficacy have changed the landscape of HCV treatment. This guideline provides updated recommendations for clinical management of HCV infection in Iran. Evidence Acquisition The recommendations of this guideline are based on international and national scientific evidences and consensus-based expert opinion. Scientific evidences were collected through a systematic review of studies that evaluated efficacy and safety of DAA regimens, using PubMed, Scopus and Web of Science. Expert opinion was based on the consensus of Iran Hepatitis Scientific Board (IHSB in the 3rd national consensus on management of Hepatitis C in Iran, held on 22nd of July 2016. Results Pegylated Interferon alpha (PegIFN, Ribavirin (RBV, Sofosbuvir (SOF, Ledipasvir (LDV and Daclatasvir (DCV are currently available in Iran. Pre-treatment assessments include HCV RNA level, HCV genotype and resistance testing, assessment of liver fibrosis, and underlying diseases. In HCV genotype 1 and 4, DCV/SOF and LDV/SOF are recommended. In HCV genotype 2, SOF plus RBV and in HCV genotype 3, DCV/SOF is recommended. Additional care for underlying diseases should be considered. Conclusions Affordable new HCV treatment regimens are available in Iran, providing an opportunity for HCV elimination. Recommendations provided in this current national guideline can facilitate evidence-based management of HCV infection.

  4. Atlas-based liver segmentation and hepatic fat-fraction assessment for clinical trials.

    Science.gov (United States)

    Yan, Zhennan; Zhang, Shaoting; Tan, Chaowei; Qin, Hongxing; Belaroussi, Boubakeur; Yu, Hui Jing; Miller, Colin; Metaxas, Dimitris N

    2015-04-01

    Automated assessment of hepatic fat-fraction is clinically important. A robust and precise segmentation would enable accurate, objective and consistent measurement of hepatic fat-fraction for disease quantification, therapy monitoring and drug development. However, segmenting the liver in clinical trials is a challenging task due to the variability of liver anatomy as well as the diverse sources the images were acquired from. In this paper, we propose an automated and robust framework for liver segmentation and assessment. It uses single statistical atlas registration to initialize a robust deformable model to obtain fine segmentation. Fat-fraction map is computed by using chemical shift based method in the delineated region of liver. This proposed method is validated on 14 abdominal magnetic resonance (MR) volumetric scans. The qualitative and quantitative comparisons show that our proposed method can achieve better segmentation accuracy with less variance comparing with two other atlas-based methods. Experimental results demonstrate the promises of our assessment framework. PMID:24962337

  5. Research on immunological safety of 20 micrograms dose of recombinant hepatitis B vaccine in adults%20微克重组乙型肝炎疫苗对成人免疫安全性的研究

    Institute of Scientific and Technical Information of China (English)

    王萍; 李艳萍; 韦琳; 农艺; 谢昌平; 杨兵华; 蓝剑

    2013-01-01

    [Objective] To observe the safety of different doses of recombinant hepatitis B vaccine (Hansenula polymorpha) in adults.[Methods] 1600 students who were over 15 years old,without hepatitis B vaccination history and contraindications to vaccination,were selected as the object of observation,while all of their hepatitis B markers,including HBsAg,anti-HBs and anti-HBc,were negative by laboratory screening,as well as ALT level were normal.By using random,double blind and the control principle,the students were divided into the experiment group and the control group,800 people in each group.According to 0,1,6 months schedule,the experiment group was given 20 μg of recombinant hepatitis B vaccine (Hansenula polymorpha) and the control group was given 10 μg of recombinant hepatitis B vaccine (Hansenula polymorpha).The venous blood samples were collected before immunization and the 4th weeks after whole course vaccination,to evaluate the immunogenicity by serum anti-HBs detection.[Results] There were 463 and 437 cases of adverse reactions in the experimental group and the control group,which the incidence rate was 57.88% and 54.63% respectively.The incidence rate of moderate and over adverse reactions was 9.25% and 8.25% respectively.The incidence rate of local reactions was 42.50% and 36.007% respectively,while the rate of local reactions over Grade 2 was 5.00% and 3.25% respectively.The incidence rate of general reactions was 37.38% and 35.50% respectively,while the rate of general reactions over Grade 2 was 4.63% and 5.63% respectively.There was no significant difference in total incidence rate of adverse reactions,incidence rate of general reactions and incidence rate of local reactions over Grade 2 between two groups.The main local adverse reactions included pain and redness,and the general reaction was mainly fever.The severe adverse reactions related to vaccination were not reported.20 micrograms dose of recombinant hepatitis B vaccine was

  6. Impact of hepatitis C virus core mutations on the response to interferon-based treatment in chronic hepatitis C

    Science.gov (United States)

    Sultana, Camelia; Oprişan, Gabriela; Teleman, Monica Delia; Dinu, Sorin; Oprea, Cristiana; Voiculescu, Mihai; Ruta, Simona

    2016-01-01

    AIM To determine whether hepatitis C virus (HCV) core substitutions play a role in the response to interferon-based treatment in Caucasian patients. METHODS One hundred eight HCV chronically infected patients initiating treatment with pegylated IFN plus ribavirin for 48 wk were tested for baseline substitutions at codons 70 and 91 of the viral core protein (BigDye Terminator vers.3.1, Applied Biosystems,) and for genetic polymorphisms in host IL28B gene rs12979860 (Custom TaqMan 5' allelic discrimination assay; Applied Biosystems). RESULTS Of the patients, all were infected with HCV genotype 1b, 44.4% had low baseline HCV viral load, and 37.9% had mild/moderate fibrosis. Only 38.9% achieved therapeutic success, defined as sustained virological response (SVR). Eighty-eight percent of the patients presented at least one substitution at core position 70 (R70Q/H) or/and position 91 (L91M). The favorable IL28B CC polymorphism was detected in only 17.6% of the patients. In the univariate analysis, young age (P < 0.001), urban residence (P = 0.004), IL28B CC genotype (P < 0.001), absence of core mutations (P = 0.005), achievement of rapid virologic response (P < 0.001) and early virological response (P < 0.001) were significantly correlated with SVR. A multivariate analysis revealed three independent predictors of therapeutic success: young age (P < 0.001), absence of core substitutions (P = 0.04) and IL28B CC genotype (P < 0.001); the model correctly classified 75.9% of SVR cases with a positive predictive value of 80.7%. CONCLUSION HCV core mutations can help distinguish between patients who can still benefit from the affordable IFN-based therapy from those who must be treated with DAAs to prevent the evolution towards end-stage liver disease. PMID:27729747

  7. Mass Spectrometry-based Quantitative Proteomic Profiling of Human Pancreatic and Hepatic Stellate Cell Lines

    OpenAIRE

    Paulo, Joao A.; Kadiyala, Vivek; Banks, Peter A; Conwell, Darwin L; Steen, Hanno

    2013-01-01

    The functions of the liver and the pancreas differ; however, chronic inflammation in both organs is associated with fibrosis. Evidence suggests that fibrosis in both organs is partially regulated by organ-specific stellate cells. We explore the proteome of human hepatic stellate cells (hHSC) and human pancreatic stellate cells (hPaSC) using mass spectrometry (MS)-based quantitative proteomics to investigate pathophysiologic mechanisms. Proteins were isolated from whole cell lysates of immorta...

  8. Shotgun metabolomic approach based on mass spectrometry for hepatic mitochondria of mice under arsenic exposure.

    Science.gov (United States)

    García-Sevillano, M A; García-Barrera, T; Navarro, F; Montero-Lobato, Z; Gómez-Ariza, J L

    2015-04-01

    Mass spectrometry (MS)-based toxicometabolomics requires analytical approaches for obtaining unbiased metabolic profiles. The present work explores the general application of direct infusion MS using a high mass resolution analyzer (a hybrid systems triple quadrupole-time-of-flight) and a complementary gas chromatography-MS analysis to mitochondria extracts from mouse hepatic cells, emphasizing on mitochondria isolation from hepatic cells with a commercial kit, sample treatment after cell lysis, comprehensive metabolomic analysis and pattern recognition from metabolic profiles. Finally, the metabolomic platform was successfully checked on a case-study based on the exposure experiment of mice Mus musculus to inorganic arsenic during 12 days. Endogenous metabolites alterations were recognized by partial least squares-discriminant analysis. Subsequently, metabolites were identified by combining MS/MS analysis and metabolomics databases. This work reports for the first time the effects of As-exposure on hepatic mitochondria metabolic pathways based on MS, and reveals disturbances in Krebs cycle, β-oxidation pathway, amino acids degradation and perturbations in creatine levels. This non-target analysis provides extensive metabolic information from mitochondrial organelle, which could be applied to toxicology, pharmacology and clinical studies.

  9. Immune responses to recombinant hepatitis B virus vaccine in human immunodeficiency virus-1-infected patients with different CD4~+ T-lymphocyte

    Institute of Scientific and Technical Information of China (English)

    蔡琳

    2014-01-01

    Objective.To compare the difference of immune responses to hepatitis B virus(HBV)vaccine in human immunodeficiency virus(HIV)-1-infected patients with different CD4+T-lymphocyte counts.Methods HIV-1 infected patients who visited clinic at the Public Health Clinical Center of Chengdu were enrolled and divided in-+

  10. Bacterial-based systems for expression and purification of recombinant Lassa virus proteins of immunological relevance

    Directory of Open Access Journals (Sweden)

    Cashman Kathleen A

    2008-06-01

    Full Text Available Abstract Background There is a significant requirement for the development and acquisition of reagents that will facilitate effective diagnosis, treatment, and prevention of Lassa fever. In this regard, recombinant Lassa virus (LASV proteins may serve as valuable tools in diverse antiviral applications. Bacterial-based systems were engineered for expression and purification of recombinant LASV nucleoprotein (NP, glycoprotein 1 (GP1, and glycoprotein 2 (GP2. Results Full-length NP and the ectodomains of GP1 and GP2 were generated as maltose-binding protein (MBP fusions in the Rosetta strains of Escherichia coli (E. coli using pMAL-c2x vectors. Average fusion protein yields per liter of culture for MBP-NP, MBP-GP1, and MBP-GP2 were 10 mg, 9 mg, and 9 mg, respectively. Each protein was captured from cell lysates using amylose resin, cleaved with Factor Xa, and purified using size-exclusion chromatography (SEC. Fermentation cultures resulted in average yields per liter of 1.6 mg, 1.5 mg, and 0.7 mg of purified NP, GP1 and GP2, respectively. LASV-specific antibodies in human convalescent sera specifically detected each of the purified recombinant LASV proteins, highlighting their utility in diagnostic applications. In addition, mouse hyperimmune ascitic fluids (MHAF against a panel of Old and New World arenaviruses demonstrated selective cross reactivity with LASV proteins in Western blot and enzyme-linked immunosorbent assay (ELISA. Conclusion These results demonstrate the potential for developing broadly reactive immunological assays that employ all three arenaviral proteins individually and in combination.

  11. Sofosbuvir-based therapy cures hepatitis C virus infection after prior treatment failures in a patient with concurrent lymphoma.

    Science.gov (United States)

    Romagnoli, Dante; Marrazzo, Alessandra; Ballestri, Stefano; Lonardo, Amedeo; Bertolotti, Marco

    2015-08-01

    We report on the first well-tolerated and successful use of sofosbuvir-based therapy in a patient in whom chronic infection with hepatitis C had preceded the development of B-cell non-Hodgkin's lymphoma. The patient had previously failed numerous attempts to clear the hepatitis C virus with traditional antiviral schedules. We demonstrate that sofosbuvir-based therapy resulted in cure of hepatitis C in a patient who had relapsed during combination therapy with an NS5A inhibitor, an NS3 protease inhibitor and ribavirin, as well as treatment failures to multiple courses of interferon-based therapy. This report also suggests that eradication of hepatitis C virus may result in the short-term prevention of B-cell non-Hodgkin's lymphoma relapse. The findings from our case require further validation in future cohorts of patients.

  12. A truncated diphtheria toxin based recombinant porcine CTLA-4 fusion toxin.

    Science.gov (United States)

    Peraino, Jaclyn Stromp; Schenk, Marian; Zhang, Huiping; Li, Guoying; Hermanrud, Christina E; Neville, David M; Sachs, David H; Huang, Christene A; Duran-Struuck, Raimon; Wang, Zhirui

    2013-05-31

    Targeted cell therapies are possible through the generation of recombinant fusion proteins that combine a toxin, such as diphtheria toxin (DT), with an antibody or other molecule that confers specificity. Upon binding of the fusion protein to the cell of interest, the diphtheria toxin is internalized which results in protein synthesis inhibition and subsequent cell death. We have recently expressed and purified the recombinant soluble porcine CTLA-4 both with and without N-glycosylation in yeast Pichia pastoris for in vivo use in our preclinical swine model. The glycosylated and non-N-glycosylated versions of this recombinant protein each bind to a porcine CD80 expressing B-cell lymphoma line (LCL13271) with equal affinity (K(D)=13 nM). In this study we have linked each of the glycosylated and non-N-glycosylated soluble porcine CTLA-4 proteins to the truncated diphtheria toxin DT390 through genetic engineering yielding three versions of the porcine CTLA-4 fusion toxins: 1) monovalent glycosylated soluble porcine CTLA-4 fusion toxin; 2) monovalent non-N-glycosylated soluble porcine CTLA-4 fusion toxin and 3) bivalent non-N-glycosylated soluble porcine CTLA-4 fusion toxin. Protein synthesis inhibition analysis demonstrated that while all three fusion toxins are capable of inhibiting protein synthesis in vitro, the non-N-glycosylated porcine CTLA-4 isoforms function most efficiently. Binding analysis using flow cytometry of the porcine CTLA-4 fusion toxins to LCL13271 cells also demonstrated that the non-N-glycosylated porcine CTLA-4 isoforms bind to these cells with higher affinity compared to the glycosylated fusion toxin. The monovalent non-N-glycosylated porcine CTLA-4 fusion toxin was tested in vivo. NSG (NOD/SCID IL-2 receptor γ(-)/(-)) mice were injected with porcine CD80(+) LCL13271 tumor cells. All animals succumbed to tumors and those treated with the monovalent non-N-glycosylated porcine CTLA-4 fusion toxin survived longer based on a symptomatic scoring

  13. Study of Domestic and Imported Recombinant Hepatitis B Vaccine Safety%国产与进口重组乙型肝炎疫苗安全性的研究

    Institute of Scientific and Technical Information of China (English)

    杨海云; 陈乐锋; 黄金凤; 梁小媚; 陈雪玲

    2015-01-01

    Objective:Understand the difference between domestic and imported vaccines the safety of both the two hepatitis B ,hepatitis Bvaccination to promote and improve immunization coverage .Methods:In Maoming City area choose healthy children aged 0~1 total 300 ,a voluntary set of principles into domestic and imported vaccines vaccine group were vaccinated with a recombinant hepatitis B vaccine and recombinant hepatitis B vaccine imported in vaccina‐tion 7 days after the follow‐up observation of systemic reactions and local swelling reactions .Results:Compared with domestic recombinant vaccine and imported recombinant hepatitis B vaccine has a similar safety profile .300 cases of re‐cipients had no local reaction ,the incidence was 3 .33% in 4 .67% ,systemic reaction ,the difference in the aspect of safety was not statistically significant (χ2 =0 .35 ,P>0 .05) .In 12 cases of adverse reactions occurred in 83 .33% (10/12) ,occurs within 1 days after inoculation ,16 .67% (2/12) occurred in 2~3 days after inoculation;the male to female ratio was 1 .40∶1 25.00% (3/12) ,occurred in inoculated first agents ,41 .67% (5/12) occurred within second agents , 33 .33% (4/12) occurred in third when the inoculation agent .Domestic vaccine group and imported vaccines group had adverse reactions in the time distribution ,population distribution ,the distribution of the inoculating needle on the same trend .Conclusion:Whether domestic or imported hepatitis B vaccine hepatitis B vaccination are part of the children after the occurrence of adverse reactions may be related to factors related to children's individual differences ,without serious consequences occur after the intervention ,the security is high ,while the domestic vaccine vaccines for national immu‐nization programs ,children free vaccinated three times ,compared with the high price of imported vaccines ,more suit‐able for the prevention and control of hepatitis .%目的:了解国产和进口两种乙肝疫苗安

  14. Study on the Failure Factors of Immunization with 10μg Recombinant Hepatitis B Vaccine(Yeast)%重组乙型肝炎疫苗(酵母)10μg成人免疫失败因素探讨

    Institute of Scientific and Technical Information of China (English)

    胡丹标; 刘世科; 赵丽丽; 黄美林; 徐爱萍; 洪因之; 曹品元

    2008-01-01

    目的 探讨重组乙型肝炎(乙肝)疫苗(酵母)[Hepatitis B Vaccine Made by Recombinant DNA Techniques in Yeast,HepB(Yeast)]10 μg成人免疫失败的因素,为制定成人HepB接种方案提供依据.方法 随机选取≥18岁易感人群,按0、1、6个月免疫程序接种HepB(Yeast)10μg,对免疫失败者进行病例对照研究.结果 成人接种10μgHepB(Yeast)免疫失败率为12.99%.免疫失败人群吸烟率、肥胖率、乙肝家族史及微量乙肝病毒(Hepatitis Bvirus,HBV)感染率均高于免疫成功人群,差异有显著的统计学意义.结论 成人接种10μgHepB(Yeast)免疫失败与吸烟、肥胖、乙肝家族史、微量HBV感染有关.

  15. [PERSPECTIVES OF DEVELOPMENT OF LIVE RECOMBINANT ANTHRAX VACCINES BASED ON OPPORTUNISTIC AND APATHOGENIC MICROORGANISMS].

    Science.gov (United States)

    Popova, P Yu; Mikshis, N I

    2016-01-01

    Live genetic engineering anthrax vaccines on the platform of avirulent and probiotic micro-organisms are a safe and adequate alternative to preparations based on attenuated Bacillus anthracis strains. Mucosal application results in a direct contact of the vaccine preparations with mucous membranes in those organs arid tissues of the macro-organisms, that are exposed to the pathogen in the first place, resulting in a development of local and systemic immune response. Live recombinant anthrax vaccines could be used both separately as well as in a prime-boost immunization scheme. The review focuses on immunogenic and protective properties of experimental live genetic engineering prearations, created based on members of geni of Salmonella, Lactobacillus and adenoviruses. PMID:27029122

  16. Human elastin-based recombinant biopolymers improve mesenchymal stem cell differentiation.

    Science.gov (United States)

    Çelebi, Betül; Cloutier, Maxime; Rabelo, Rodrigo B; Balloni, Rodrigo; Mantovani, Diego; Bandiera, Antonella

    2012-11-01

    Elastin-based polypeptides are a class of smart biopolymers representing an important model in the design of biomaterials. The combination of biomimetic materials with cells that have great plasticity provides a promising strategy for the realization of highly engineered cell-based constructs for regenerative medicine and tissue repair applications. Two recombinant biopolymers inspired by human elastin are assessed as coating agents to prepare biomimetic surfaces for cell culture. These substrates are assayed for hBM MSC culture. The coated surfaces are also characterized with AFM to evaluate the topographical features of the deposited biopolymers. The results suggest that the elastin-derived biomimetic surfaces play a stimulatory role on osteogenic differentiation of MSCs.

  17. [PERSPECTIVES OF DEVELOPMENT OF LIVE RECOMBINANT ANTHRAX VACCINES BASED ON OPPORTUNISTIC AND APATHOGENIC MICROORGANISMS].

    Science.gov (United States)

    Popova, P Yu; Mikshis, N I

    2016-01-01

    Live genetic engineering anthrax vaccines on the platform of avirulent and probiotic micro-organisms are a safe and adequate alternative to preparations based on attenuated Bacillus anthracis strains. Mucosal application results in a direct contact of the vaccine preparations with mucous membranes in those organs arid tissues of the macro-organisms, that are exposed to the pathogen in the first place, resulting in a development of local and systemic immune response. Live recombinant anthrax vaccines could be used both separately as well as in a prime-boost immunization scheme. The review focuses on immunogenic and protective properties of experimental live genetic engineering prearations, created based on members of geni of Salmonella, Lactobacillus and adenoviruses.

  18. Comparison of three different recombinant hepatitis B vaccines: GeneVac-B, Engerix B and Shanvac B in high risk infants born to HBsAg positive mothers in India

    Institute of Scientific and Technical Information of China (English)

    Vijayakumar Velu; Subhadra Nandakumar; Saravanan Shanmugam; Suresh Sakharam Jadhav; Prasad Suryakant Kulkarni; Sadras Panchatcharam Thyagarajan

    2007-01-01

    AIM: To evaluate a low cost Indian recombinant hepatitis B vaccine GeneVac-B for its immunogenicity and safety in comparison to Engerix B and Shanvac B vaccine in high risk newborn infants born to (hepatitis B surface antigen) HBsAg positive mothers.METHODS: A total of 158 infants were enrolled in the study. Fifty eight infants were enrolled in the GeneVac-B group while 50 each were included for Engerix B and Shanvac B groups. A three-dose regimen of vaccination; at birth (within 24 h of birth), 1st mo and 6 mo. were adopted with 10 ng dosage administered uniformly in all the three groups. Clinical and immunological parameters were assessed for safety and immunogenicity of the vaccines, in all the enrolled infants.RESULTS: Successful follow up until seven months of age was achieved in 83% (48/58) for GeneVac-B, 76% (38/50) and 64% (32/50) for Engerix B and Shanvac B groups respectively. 100% seroconversion and seroprotection was achieved in all the three groups of infants. The geometric mean titers of anti-HBs one month after the completion of three dose of vaccination were 90.5, 80.9 and 72.5 mlU/mL in GeneVac-B, Engerix B and Shanvac B vaccine group respectively. Furthermore the level of anti-HBs increases with age of babies who were born to HBsAg positive mothers. The GMT values of anti-HBs were 226.7, 193.9 and 173.6 mlU/mL respectively in GeneVac-B, Engerix B and Shanvac B groups one year after the completion of the three doses of vaccine. No systemic reactions were reported in infants during the entire vaccination process of GeneVac-B and the other two vaccines. Clinical safety parameters remained within the normal limits throughout the study period.CONCLUSION: The study concludes that there is no significant difference between the three recombinant hepatitis B vaccines. Administration of these vaccines within 24 h of birth to babies, born to HBsAg positive mothers will reduce the incidence of HBV infection.

  19. Hepatitis B prevalence and incidence in Greenland: a population-based cohort study.

    Science.gov (United States)

    Børresen, Malene Landbo; Andersson, Mikael; Wohlfahrt, Jan; Melbye, Mads; Biggar, Robert J; Ladefoged, Karin; Panum, Inge; Koch, Anders

    2015-03-15

    Greenland remains a highly endemic area for hepatitis B virus (HBV) infection. This is in sharp contrast to other modern societies, such as Denmark. To address this discrepancy, we investigated the natural history of HBV infection in Greenland by estimating the age-specific incidence of HBV infection, the proportion of chronic carriers, and the rates of hepatitis B surface antigen seroclearance. In total, 8,879 Greenlanders (16% of the population) from population-based surveys conducted in 1987 and 1998 were followed through March 2010. Data on HBV status were supplemented by HBV test results from all available HBV registries in Greenland to determine changes in HBV status over time. Incidence rates of HBV infection and hepatitis B surface antigen seroclearance were estimated after taking into account interval censoring. The incidence of HBV infection in 5-14-year-old subjects was less than 1 per 100 person-years and peaked at 5 per 100 person-years in persons 15-24 years of age. Overall, 17.5% of persons infected in adulthood were estimated to become chronic carriers. HBV is primarily transmitted in adolescence and adulthood in Greenland. In contrast to what is observed in most other populations, HBV-infected adults in Greenland have a high risk of progressing to chronic HBV carriage. This phenomenon might explain how the high rate of infection is maintained in Greenland.

  20. Stability of PIKA Recombinant Hepatitis B Vaccine(Hansenula polymorpha)%皮卡重组乙型肝炎疫苗(汉逊酵母)的稳定性

    Institute of Scientific and Technical Information of China (English)

    李晓宇; 高俊清; 张燕; 金贞姬; 刘照惠

    2011-01-01

    Objective To observe the stability of PIKA recombinant hepatitis B (HB) vaccine (Hansenula polymorpha).Methods PIKA recombinant HB vaccine (H. polymorpha) was stored at 37, 25 and 2 ~ 8℃ respectively for various days, then determined for relative potency (RP) in vitro, pH value and purity, and observed for stability. Results The RP of vaccine decresed slightly after storage at 37℃ for 45 d but still met the relevant requirements. Neither RP nor pH value showed signficant change after storage at 25℃ for 6 months. However, after storage at 2 ~ 8℃ for 12 months, no signifcant change was observed in RP, pH value or purity of the vaccine. Conclusion PIKA recombinant HB vaccine (H. polymorpha) showed high stability.%目的 观察皮卡重组乙型肝炎疫苗(汉逊酵母)的稳定性.方法 将皮卡重组乙型肝炎疫苗(汉逊酵母)分别于37、25和2-8℃放置不同时间,检测疫苗的体外相对效力(Relative potency,RP)、pH值和纯度的变化,观察其稳定性.结果 疫苗于37℃放置45 d,RP略有下降,但仍在合格范围内;25℃放置6个月,RP和pH值均无明显变化;2-8℃放置12个月,RP、pH值和纯度均无明显变化.结论 皮卡重组乙型肝炎疫苗(汉逊酵母)稳定性良好.

  1. Developing a novel hepatitis B core – based antigen presentation system

    OpenAIRE

    Peyret, Hadrien

    2014-01-01

    Plant-produced proteins of pharmaceutical interest are beginning to reach the market, and the advantages of transient plant expression systems are gaining increasing recognition. In parallel, the use of virus-like particles (VLPs) has become standard in vaccine design. The pEAQ-HT vector derived from the cowpea mosaic virus – based CPMV-HT expression system has been shown to allow the production of large amounts of recombinant proteins, including VLPs, in Nicotiana benthamiana. Moreover, prev...

  2. Liver stiffness measurement-based scoring system for significant inflammation related to chronic hepatitis B.

    Directory of Open Access Journals (Sweden)

    Mei-Zhu Hong

    Full Text Available Liver biopsy is indispensable because liver stiffness measurement alone cannot provide information on intrahepatic inflammation. However, the presence of fibrosis highly correlates with inflammation. We constructed a noninvasive model to determine significant inflammation in chronic hepatitis B patients by using liver stiffness measurement and serum markers.The training set included chronic hepatitis B patients (n = 327, and the validation set included 106 patients; liver biopsies were performed, liver histology was scored, and serum markers were investigated. All patients underwent liver stiffness measurement.An inflammation activity scoring system for significant inflammation was constructed. In the training set, the area under the curve, sensitivity, and specificity of the fibrosis-based activity score were 0.964, 91.9%, and 90.8% in the HBeAg(+ patients and 0.978, 85.0%, and 94.0% in the HBeAg(- patients, respectively. In the validation set, the area under the curve, sensitivity, and specificity of the fibrosis-based activity score were 0.971, 90.5%, and 92.5% in the HBeAg(+ patients and 0.977, 95.2%, and 95.8% in the HBeAg(- patients. The liver stiffness measurement-based activity score was comparable to that of the fibrosis-based activity score in both HBeAg(+ and HBeAg(- patients for recognizing significant inflammation (G ≥3.Significant inflammation can be accurately predicted by this novel method. The liver stiffness measurement-based scoring system can be used without the aid of computers and provides a noninvasive alternative for the prediction of chronic hepatitis B-related significant inflammation.

  3. Liver Stiffness Measurement-Based Scoring System for Significant Inflammation Related to Chronic Hepatitis B

    Science.gov (United States)

    Hong, Mei-Zhu; Zhang, Ru-Mian; Chen, Guo-Liang; Huang, Wen-Qi; Min, Feng; Chen, Tian; Xu, Jin-Chao; Pan, Jin-Shui

    2014-01-01

    Objectives Liver biopsy is indispensable because liver stiffness measurement alone cannot provide information on intrahepatic inflammation. However, the presence of fibrosis highly correlates with inflammation. We constructed a noninvasive model to determine significant inflammation in chronic hepatitis B patients by using liver stiffness measurement and serum markers. Methods The training set included chronic hepatitis B patients (n = 327), and the validation set included 106 patients; liver biopsies were performed, liver histology was scored, and serum markers were investigated. All patients underwent liver stiffness measurement. Results An inflammation activity scoring system for significant inflammation was constructed. In the training set, the area under the curve, sensitivity, and specificity of the fibrosis-based activity score were 0.964, 91.9%, and 90.8% in the HBeAg(+) patients and 0.978, 85.0%, and 94.0% in the HBeAg(−) patients, respectively. In the validation set, the area under the curve, sensitivity, and specificity of the fibrosis-based activity score were 0.971, 90.5%, and 92.5% in the HBeAg(+) patients and 0.977, 95.2%, and 95.8% in the HBeAg(−) patients. The liver stiffness measurement-based activity score was comparable to that of the fibrosis-based activity score in both HBeAg(+) and HBeAg(−) patients for recognizing significant inflammation (G ≥3). Conclusions Significant inflammation can be accurately predicted by this novel method. The liver stiffness measurement-based scoring system can be used without the aid of computers and provides a noninvasive alternative for the prediction of chronic hepatitis B-related significant inflammation. PMID:25360742

  4. Development of Ss-NIE-1 recombinant antigen based assays for immunodiagnosis of strongyloidiasis.

    Science.gov (United States)

    Rascoe, Lisa N; Price, Courtney; Shin, Sun Hee; McAuliffe, Isabel; Priest, Jeffrey W; Handali, Sukwan

    2015-04-01

    Strongyloides stercoralis is a widely distributed parasite that infects 30 to 100 million people worldwide. In the United States strongyloidiasis is recognized as an important infection in immigrants and refugees. Public health and commercial reference laboratories need a simple and reliable method for diagnosis of strongyloidiasis to identify and treat cases and to prevent transmission. The recognized laboratory test of choice for diagnosis of strongyloidiasis is detection of disease specific antibodies, most commonly using a crude parasite extract for detection of IgG antibodies. Recently, a luciferase tagged recombinant protein of S. stercoralis, Ss-NIE-1, has been used in a luciferase immunoprecipitation system (LIPS) to detect IgG and IgG4 specific antibodies. To promote wider adoption of immunoassays for strongyloidiasis, we used the Ss-NIE-1 recombinant antigen without the luciferase tag and developed ELISA and fluorescent bead (Luminex) assays to detect S. stercoralis specific IgG4. We evaluated the assays using well-characterized sera from persons with or without presumed strongyloidiasis. The sensitivity and specificity of Ss-NIE-1 IgG4 ELISA were 95% and 93%, respectively. For the IgG4 Luminex assay, the sensitivity and specificity were 93% and 95%, respectively. Specific IgG4 antibody decreased after treatment in a manner that was similar to the decrease of specific IgG measured in the crude IgG ELISA. The sensitivities of the Ss-NIE-1 IgG4 ELISA and Luminex assays were comparable to the crude IgG ELISA but with improved specificities. However, the Ss-NIE-1 based assays are not dependent on native parasite materials and can be performed using widely available laboratory equipment. In conclusion, these newly developed Ss-NIE-1 based immunoassays can be readily adopted by public health and commercial reference laboratories for routine screening and clinical diagnosis of S. stercoralis infection in refugees and immigrants in the United States.

  5. Copy-number gains of HUWE1 due to replication- and recombination-based rearrangements.

    Science.gov (United States)

    Froyen, Guy; Belet, Stefanie; Martinez, Francisco; Santos-Rebouças, Cíntia Barros; Declercq, Matthias; Verbeeck, Jelle; Donckers, Lene; Berland, Siren; Mayo, Sonia; Rosello, Monica; Pimentel, Márcia Mattos Gonçalves; Fintelman-Rodrigues, Natalia; Hovland, Randi; Rodrigues dos Santos, Suely; Raymond, F Lucy; Bose, Tulika; Corbett, Mark A; Sheffield, Leslie; van Ravenswaaij-Arts, Conny M A; Dijkhuizen, Trijnie; Coutton, Charles; Satre, Veronique; Siu, Victoria; Marynen, Peter

    2012-08-10

    We previously reported on nonrecurrent overlapping duplications at Xp11.22 in individuals with nonsyndromic intellectual disability (ID) harboring HSD17B10, HUWE1, and the microRNAs miR-98 and let-7f-2 in the smallest region of overlap. Here, we describe six additional individuals with nonsyndromic ID and overlapping microduplications that segregate in the families. High-resolution mapping of the 12 copy-number gains reduced the minimal duplicated region to the HUWE1 locus only. Consequently, increased mRNA levels were detected for HUWE1, but not HSD17B10. Marker and SNP analysis, together with identification of two de novo events, suggested a paternally derived intrachromosomal duplication event. In four independent families, we report on a polymorphic 70 kb recurrent copy-number gain, which harbors part of HUWE1 (exon 28 to 3' untranslated region), including miR-98 and let-7f-2. Our findings thus demonstrate that HUWE1 is the only remaining dosage-sensitive gene associated with the ID phenotype. Junction and in silico analysis of breakpoint regions demonstrated simple microhomology-mediated rearrangements suggestive of replication-based duplication events. Intriguingly, in a single family, the duplication was generated through nonallelic homologous recombination (NAHR) with the use of HUWE1-flanking imperfect low-copy repeats, which drive this infrequent NAHR event. The recurrent partial HUWE1 copy-number gain was also generated through NAHR, but here, the homologous sequences used were identified as TcMAR-Tigger DNA elements, a template that has not yet been reported for NAHR. In summary, we showed that an increased dosage of HUWE1 causes nonsyndromic ID and demonstrated that the Xp11.22 region is prone to recombination- and replication-based rearrangements.

  6. Estimation of Charge Exchange Recombination Emission Based on Diagnostic Neutral Beam on the Experimental Advanced Superconducting Tokamak

    Institute of Scientific and Technical Information of China (English)

    ZHANG Xian-Mei; WAN Bao-Nian; WU Zhen-Wei

    2007-01-01

    Diagnostic neutral beam (DNB) attenuation and charge exchange recombination emission are estimated on EAST tokamak. Approximately 40% of the beam with the energy of 50 keV can reach the plasma centre (r = 0) for the typical parameters of the Experimental Advanced Superconducting Tokamak (EAST) plasma. Emissivities of CVI (n = 8 → 7, 529.0nm) and OVⅢ (n = 10 → 9, 607.0 nm) visible charge exchange recombination emissions based on the DNB are estimated. The emissivities of the visible bremsstrahlung emission near this wavelength are also calculated for comparison. The results show that the charge exchange recombination emission is about two orders of magnitude greater than the bremsstrahlung emission. It is theoretically indicated that the ratio of signal of charge exchange recombination spectroscopy to the noise from background bremsstrahlung emission,S/N, is large enough in the EAST tokamak with the typical designed parameters. The present results are helpful for experiment design of charge-exchange recombination spectroscopy based on the DNB in the EAST tokamak.

  7. Establishment and stability of seed bank of recombinant hepatitis E vaccine%重组戊型肝炎疫苗菌种库的建立及其稳定性

    Institute of Scientific and Technical Information of China (English)

    曹玉锋; 王伟善; 徐军; 时成波; 孟多佳; 迟春萍; 王玉梅; 贾媛; 张剑锋; 常东英

    2012-01-01

    Objective To establish three-tier seed bank of recombinant hepatitis E vaccine and study its stability. Methods Recombinant E. coli HEV179/BL21 (DE3) was resuscitated, activated and subjected to overall control tests, and the qualified ones were lyophilized as primary seed bank. The seeds from primary seed bank were subcultured, propagated and subjected to overall control tests, while the qualified ones were lyophilized as master seed bank. The seeds from master seed bank were subcultured, propagated and subjected to overall tests, while the qualified ones were cryopreserved in glycerol and served as working seed bank. The three-tier seed bank was subjected to overall control tests, and the recombinant strain was evaluated for stabilities after lyophilization and cryopreservation in glycerol as well as the genetic stability. Results The established three-tier seed bank was qualified in overall control tests, of which the stabilities after lyophilization and cryopreservation in glycerol as well as the genetic stability met the relevant requirements. The growth characteristics as well as genetic and expression stabilities of recombinant strain showed no significant change. Conclusion Three-tier seed bank of HEV179 / BL2l(DE3) was successfully established, and the seeds showed high stability and was suitable for industrial production of recombinant hepatitis E vaccine.%目的 建立重组戊型肝炎疫苗工程菌三级种子库,并分析其稳定性.方法 将工程菌株HEV179/BL21 (DE3)复苏活化,经全面检测合格后冻干保存即为原始种子库;原始种子库传代扩增,经全面检定合格后,冻干保存即为主种子库;主种子库经传代扩增和全面检定合格后,甘油冻存即为工作种子库.对三级种子库进行全面检定,并对工程菌进行冻干及甘油冻存稳定性、甘油冻存时间稳定性及质粒稳定性检测.结果 建立的三级种子库经全面检定合格;菌种冻干稳定性、甘油冻存稳定

  8. Hepatic trauma: CT findings and considerations based on our experience in emergency diagnostic imaging

    Energy Technology Data Exchange (ETDEWEB)

    Romano, Luigia; Giovine, Sabrina; Guidi, Guido; Tortora, Giovanni; Cinque, Teresa; Romano, Stefania E-mail: stefromano@libero.it

    2004-04-01

    Abdominal blunt trauma represents the main cause of death in people of age less than 40 years; the liver injury occurs frequently, with an incidence varying from 3 to 10%. Isolated hepatic lesions are rare and in 77-90% of cases, lesions of other organs and viscera are involved. Right hepatic lobe is a frequent site of injury, because it is the more voluminous portion of liver parenchyma; posterior superior hepatic segments are proximal to fixed anatomical structures such as ribs and spine that may have an important role in determining of the lesion. The coronal ligaments' insertion in this parenchymal region augments the effect of acceleration-deceleration mechanism. Associated lesions usually are homolateral costal fractures, laceration or contusion of the inferior right pulmonary lobe, haemothorax, pneumothorax, renal and/or adrenal lesions. Traumatic lesions of left hepatic lobe are rare and usually associated with direct impact on the superior abdomen, such as in car-crash when the wheel causes a compressive effect on thorax and abdomen. Associated lesions to left hepatic lobe injuries correlated to this mechanism are: sternal fractures, pancreatic, myocardial, gastrointestinal tract injuries. Lesions of the caudal lobe are extremely rare, usually not isolated and noted with other large parenchymal lesions. The Institution of Specialized Trauma Centers and the technical progress in imaging methodology developed in the last years a great reduction of mortality. New diagnostic methodologies allow a reduction of negatives laparotomies and allow the possibility of conservative treatment of numerous traumatic lesions; however, therapy depends from imaging findings and clinical conditions of the patient. Computed tomography (CT) certainly presents a large impact on diagnosis and management of patients with lesions from blunt abdominal traumas. It is important to establish a prognostic criteria allowing decisions for conservative or surgical treatment; CT

  9. Hepatic trauma: CT findings and considerations based on our experience in emergency diagnostic imaging

    International Nuclear Information System (INIS)

    Abdominal blunt trauma represents the main cause of death in people of age less than 40 years; the liver injury occurs frequently, with an incidence varying from 3 to 10%. Isolated hepatic lesions are rare and in 77-90% of cases, lesions of other organs and viscera are involved. Right hepatic lobe is a frequent site of injury, because it is the more voluminous portion of liver parenchyma; posterior superior hepatic segments are proximal to fixed anatomical structures such as ribs and spine that may have an important role in determining of the lesion. The coronal ligaments' insertion in this parenchymal region augments the effect of acceleration-deceleration mechanism. Associated lesions usually are homolateral costal fractures, laceration or contusion of the inferior right pulmonary lobe, haemothorax, pneumothorax, renal and/or adrenal lesions. Traumatic lesions of left hepatic lobe are rare and usually associated with direct impact on the superior abdomen, such as in car-crash when the wheel causes a compressive effect on thorax and abdomen. Associated lesions to left hepatic lobe injuries correlated to this mechanism are: sternal fractures, pancreatic, myocardial, gastrointestinal tract injuries. Lesions of the caudal lobe are extremely rare, usually not isolated and noted with other large parenchymal lesions. The Institution of Specialized Trauma Centers and the technical progress in imaging methodology developed in the last years a great reduction of mortality. New diagnostic methodologies allow a reduction of negatives laparotomies and allow the possibility of conservative treatment of numerous traumatic lesions; however, therapy depends from imaging findings and clinical conditions of the patient. Computed tomography (CT) certainly presents a large impact on diagnosis and management of patients with lesions from blunt abdominal traumas. It is important to establish a prognostic criteria allowing decisions for conservative or surgical treatment; CT findings

  10. Seroprevalence of Hepatitis B surface antigen, antibodies to the Hepatitis C virus, and human immunodeficiency virus in a hospital-based population in Jaipur, Rajasthan

    Directory of Open Access Journals (Sweden)

    Sood Smita

    2010-01-01

    Full Text Available Background: Hepatitis B, hepatitis C, and HIV infections are a serious global and public health problem. To assess the magnitude and dynamics of disease transmission and for its prevention and control, the study of its seroprevalence is important. A private hospital catering to the needs of a large population represents an important center for serological surveys. Available data, at Rajasthan state level, on the seroprevalence of these bloodborne pathogens is also very limited. Objective: A study was undertaken to estimate the seroprevalence of hepatitis B surface antigen (HBsAg and antibodies to hepatitis C (anti-HCV Ab and human immunodeficiency virus (anti-HIV Ab in both the sexes and different age groups in a hospital-based population in Jaipur, Rajasthan. Materials and Methods: Serum samples collected over a period of 14 months from patients attending OPDs and admitted to various IPDs of Fortis Escorts Hospital, Jaipur, were subjected within the hospital-based lab for the detection of HBsAg and anti-HCV Ab and anti-HIV Ab using rapid card tests. This was followed by further confirmation of all reactive samples by a microparticle enzyme immunoassay (Abbott AxSYM at Super Religare Laboratories (formerly SRL Ranbaxy Reference Lab, Mumbai. Results: The seroprevalence of HBsAg was found to be 0.87%, of anti-HCV Ab as 0.28%, and of anti-HIV Ab as 0.35%. Conclusion: The study throws light on the magnitude of viral transmission in the community in the state of Rajasthan and provides a reference for future studies.

  11. The new pLAI (lux regulon based auto-inducible expression system for recombinant protein production in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Nocadello Salvatore

    2012-01-01

    Full Text Available Abstract Background After many years of intensive research, it is generally assumed that no universal expression system can exist for high-level production of a given recombinant protein. Among the different expression systems, the inducible systems are the most popular for their tight regulation. However, induction is in many cases less favorable due to the high cost and/or toxicity of inducers, incompatibilities with industrial scale-up or detrimental growth conditions. Expression systems using autoinduction (or self-induction prove to be extremely versatile allowing growth and induction of recombinant proteins without the need to monitor cell density or add inducer. Unfortunately, almost all the actual auto inducible expression systems need endogenous or induced metabolic changes during the growth to trigger induction, both frequently linked to detrimental condition to cell growth. In this context, we use a simple modular approach for a cell density-based genetic regulation in order to assemble an autoinducible recombinant protein expression system in E. coli. Result The newly designed pLAI expression system places the expression of recombinant proteins in Escherichia coli under control of the regulatory genes of the lux regulon of Vibrio fischeri's Quorum Sensing (QS system. The pLAI system allows a tight regulation of the recombinant gene allowing a negligible basal expression and expression only at high cell density. Sequence optimization of regulative genes of QS of V. fischeri for expression in E. coli upgraded the system to high level expression. Moreover, partition of regulative genes between the plasmid and the host genome and introduction of a molecular safety lock permitted tighter control of gene expression. Conclusion Coupling gene expression to cell density using cell-to-cell communication provides a promising approach for recombinant protein production. The system allows the control of expression of the target recombinant gene

  12. Development of recombinant nucleoprotein-based diagnostic systems for Lassa fever.

    Science.gov (United States)

    Saijo, Masayuki; Georges-Courbot, Marie-Claude; Marianneau, Philippe; Romanowski, Victor; Fukushi, Shuetsu; Mizutani, Tetsuya; Georges, Alain-Jean; Kurata, Takeshi; Kurane, Ichiro; Morikawa, Shigeru

    2007-09-01

    Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa cells by transfection with the expression vector encoding cDNA of the LASV-NP gene. An immunoglobulin G enzyme-linked immunosorbent assay (ELISA) using LASV-rNP and an indirect immunofluorescence assay using LASV-rNP-expressing HeLa cells were confirmed to have high sensitivity and specificity in the detection of LASV-antibodies. A novel monoclonal antibody to LASV-rNP, monoclonal antibody 4A5, was established. A sandwich antigen capture (Ag-capture) ELISA using the monoclonal antibody and an anti-LASV-rNP rabbit serum as capture and detection antibodies, respectively, was then developed. Authentic LASV nucleoprotein in serum samples collected from hamsters experimentally infected with LASV was detected by the Ag-capture ELISA. The Ag-capture ELISA specifically detected LASV-rNP but not the rNPs of lymphocytic choriomeningitis virus or Junin virus. The sensitivity of the Ag-capture ELISA in detecting LASV antigens was comparable to that of reverse transcription-PCR in detecting LASV RNA. These LASV rNP-based diagnostics were confirmed to be useful in the diagnosis of LF even in institutes without a high containment laboratory, since the antigens can be prepared without manipulation of the infectious viruses. PMID:17634509

  13. Development of Recombinant Nucleoprotein-Based Diagnostic Systems for Lassa Fever▿

    Science.gov (United States)

    Saijo, Masayuki; Georges-Courbot, Marie-Claude; Marianneau, Philippe; Romanowski, Victor; Fukushi, Shuetsu; Mizutani, Tetsuya; Georges, Alain-Jean; Kurata, Takeshi; Kurane, Ichiro; Morikawa, Shigeru

    2007-01-01

    Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa cells by transfection with the expression vector encoding cDNA of the LASV-NP gene. An immunoglobulin G enzyme-linked immunosorbent assay (ELISA) using LASV-rNP and an indirect immunofluorescence assay using LASV-rNP-expressing HeLa cells were confirmed to have high sensitivity and specificity in the detection of LASV-antibodies. A novel monoclonal antibody to LASV-rNP, monoclonal antibody 4A5, was established. A sandwich antigen capture (Ag-capture) ELISA using the monoclonal antibody and an anti-LASV-rNP rabbit serum as capture and detection antibodies, respectively, was then developed. Authentic LASV nucleoprotein in serum samples collected from hamsters experimentally infected with LASV was detected by the Ag-capture ELISA. The Ag-capture ELISA specifically detected LASV-rNP but not the rNPs of lymphocytic choriomeningitis virus or Junin virus. The sensitivity of the Ag-capture ELISA in detecting LASV antigens was comparable to that of reverse transcription-PCR in detecting LASV RNA. These LASV rNP-based diagnostics were confirmed to be useful in the diagnosis of LF even in institutes without a high containment laboratory, since the antigens can be prepared without manipulation of the infectious viruses. PMID:17634509

  14. Detection of Hepatitis B Virus DNA by Duplex Scorpion Primer-based PCR Assay

    Institute of Scientific and Technical Information of China (English)

    KONG De-Ming孔德明; SHEN Han-Xi沈含熙; MI Huai-Feng宓怀风

    2004-01-01

    The application of a new fiuorogenic probe-based PCR assay (PCR duplex scorpion primer assay) to the detection of Hepatitis B virus (HBV) DNA in human sera was described. Duplex scorpion primer is a modified variant of duplex Amplifluor, and the incorporation of a PCR stopper between probe and primer sequences improve the detection specificity and sensitivity. Combined with PCR amplification, this probe can give unambiguous positive results for the reactions initiated with more than 20 HBV molecules. In addition, the particular unimolecular probing mechanism of this probe makes the use of short target-specific probe sequence possible, which will render this probe applicable in some specific systems.

  15. Live Attenuated Vaccine Based on Duck Enteritis Virus against Duck Hepatitis A Virus Types 1 and 3

    Science.gov (United States)

    Zou, Zhong; Ma, Ji; Huang, Kun; Chen, Huanchun; Liu, Ziduo; Jin, Meilin

    2016-01-01

    As causative agents of duck viral hepatitis, duck hepatitis A virus type 1 (DHAV-1) and type 3 (DHAV-3) causes significant economic losses in the duck industry. However, a licensed commercial vaccine that simultaneously controls both pathogens is currently unavailable. Here, we generated duck enteritis virus recombinants (rC-KCE-2VP1) containing both VP1 from DHAV-1 (VP1/DHAV-1) and VP1 from DHAV-3 (VP1/DHAV-3) between UL27 and UL26. A self-cleaving 2A-element of FMDV was inserted between the two different types of VP1, allowing production of both proteins from a single open reading frame. Immunofluorescence and Western blot analysis results demonstrated that both VP1 proteins were robustly expressed in rC-KCE-2VP1-infected chicken embryo fibroblasts. Ducks that received a single dose of rC-KCE-2VP1 showed potent humoral and cellular immune responses and were completely protected against challenges of both pathogenic DHAV-1 and DHAV-3 strains. The protection was rapid, achieved as early as 3 days after vaccination. Moreover, viral replication was fully blocked in vaccinated ducks as early as 1 week post-vaccination. These results demonstrated, for the first time, that recombinant rC-KCE-2VP1 is potential fast-acting vaccine against DHAV-1 and DHAV-3. PMID:27777571

  16. Serodiagnosis of Rift Valley fever in African wildlife using a recombinant nucleocapsid-based indirect ELISA

    International Nuclear Information System (INIS)

    Antibodies to Rift Valley fever virus (RVFV) have been found in many wildlife species but their importance in the epidemiology of the disease during the inter-epidemic and epidemic periods, and including their possible specific role in the cryptic maintenance of the virus is not elucidated. A recently developed indirect ELISA (I-ELISA) based on recombinant nucleocapsid protein (rNp) of RVF virus was reported to have high analytical accuracy for the detection of IgG antibody in African buffalo sera. An indirect ELISA (I-ELISA) based on the recombinant nucleocapsid protein (rNp) of Rift Valley fever virus (RVFV) was evaluated for the detection of specific serum IgG antibody in African wildlife. Data sets derived from field-collected sera (n = 877) in Africa (antelopes = 529, black rhinoceros = 43, common zebra = 24, elephant = 73, giraffe = 81, grevy zebra = 78, warthog = 49) were categorized according to the results of a virus neutralization test. Dose response curves using different dilutions of sera known to be positive or negative in the virus neutralisation test had the expected analytical slope and the I-ELISA clearly differentiated between different levels of specific IgG antibody against RVFV in African wildlife. At cut-offs optimised by the two-graph receiver operating characteristics analysis, the diagnostic sensitivity of the I-ELISA was 100% and diagnostic specificity ranged from 99.8% to 100% while estimates for the Youden's index (J) and efficiency (Ef) ranged from 0.99 to 1 and from 99.7% to 100%, respectively. The rNp-based I-ELISA is highly accurate, safe, and offers a single assay format for rapid detection of IgG antibody to RVFV in sera of different wildlife species. This study confirm previous findings that the rNp-based I-ELISA accurately identifies sera with different concentrations of specific IgG antibodies to RVF virus, and compared to virus neutralization test it has very high diagnostic performance in various wildlife animal species. As a

  17. 干扰素用于重组乙肝疫苗的佐剂作用%Interferon as An Adjuvant of Recombinant Hepatitis B Vaccine

    Institute of Scientific and Technical Information of China (English)

    罗璇; 魏自力; 陈万革; 蔡岩; 刘大维; 韩立卓

    2000-01-01

    Objective To improve the immuogenicity of hepatitis B vaccine. Methods Mice wereimmunized with the hepatitis B vaccine using interferon and Al(OH)3 complex adjuvant,and the humoral andcell immune responses of them were studied. Results Both humoral and cell immunity of the vaccine usingcomplex adjuvant were significantly higher than that using Al(OH)3 adjuvant alone. Conclusion Interferoncan be used as an adjuvant of hapatitis B vaccine.%目的 为了提高乙肝疫苗的免疫原性。方法 在重组乙肝疫苗中加入不同单位的干扰素构成复合佐剂,通过小鼠腹腔和肌肉两种免疫途径,对疫苗的体液免疫和细胞免疫进行了检测。结果 干扰素作为复合铝佐剂,无论是对体液免疫还是细胞免疫,效果均较对照明显提高。结论 不同亚型的IFN作为乙肝疫苗佐剂,均可提高疫苗的免疫原性。

  18. IL-12-based vaccination therapy reverses liver-induced systemic tolerance in a mouse model of hepatitis B virus carrier.

    Science.gov (United States)

    Zeng, Zhutian; Kong, Xiaohui; Li, Fenglei; Wei, Haiming; Sun, Rui; Tian, Zhigang

    2013-10-15

    Liver-induced systemic immune tolerance that occurs during chronic hepadnavirus infection is the biggest obstacle for effective viral clearance. Immunotherapeutic reversal of this tolerance is a promising strategy in the clinic but remains to be explored. In this study, using a hepatitis B virus (HBV)-carrier mouse model, we report that IL-12-based vaccination therapy can efficiently reverse systemic tolerance toward HBV. HBV-carrier mice lost responsiveness to hepatitis B surface Ag (HBsAg) vaccination, and IL-12 alone could not reverse this liver-induced immune tolerance. However, after IL-12-based vaccination therapy, the majority of treated mice became HBsAg(-) in serum; hepatitis B core Ag was also undetectable in hepatocytes. HBV clearance was dependent on HBsAg vaccine-induced anti-HBV immunity. Further results showed that IL-12-based vaccination therapy strongly enhanced hepatic HBV-specific CD8(+) T cell responses, including proliferation and IFN-γ secretion. Systemic HBV-specific CD4(+) T cell responses were also restored in HBV-carrier mice, leading to the arousal of HBsAg-specific follicular Th-germinal center B cell responses and anti-hepatitis B surface Ag Ab production. Recovery of HBsAg-specific responses also correlated with both reduced CD4(+)Foxp3(+) regulatory T cell frequency and an enhanced capacity of effector T cells to overcome inhibition by regulatory T cells. In conclusion, IL-12-based vaccination therapy may reverse liver-induced immune tolerance toward HBV by restoring systemic HBV-specific CD4(+) T cell responses, eliciting robust hepatic HBV-specific CD8(+) T cell responses, and facilitating the generation of HBsAg-specific humoral immunity; thus, this therapy may become a viable approach to treating patients with chronic hepatitis B. PMID:24048897

  19. Antibody reaction to immunization with recombinant hepatitis B vaccine in undergraduates%大学生乙型肝炎基因重组疫苗接种效果分析

    Institute of Scientific and Technical Information of China (English)

    黄茵; 陈智; 蔡玲斐

    2001-01-01

    Objective To evaluate the safety and immunogenicity of recombinant hepatitis B(HB)vaccine to undergraduates and lay a basis for formulaing an immunization strategy.Method Yeast derived recombinant HB vaccine(YDV)was given at 0,1 and 2 months to 165 undergraduates whose hepatitis B virus markers (HBVM) were negative before vaccination,and then to detect the titer of antibody against hepatitis B surface antigen(anti-HBs)with radioimmunoassay(RIA).Result No severe adverse reaction was found in all vaccinee.Positive seroconvertion rate was 81.41 percent two months after three-dose schedule,but most subjects were lowresponders,whose mean geometric titer(GMT)were at 10-99IU/L.Only 1.92 percent of the subjects reached the level that is higher than 100IU/L.7 of 12 non-responders to primary HB vaccination showed response to HB antigen after our 3-dose schedule.Conclusion Safety and immunogenicity of recombinant HB vaccine is good for undergraduates,but the levels of anti-HBs are too low.This response seems not to be influenced by sex.Non-responders to primary HB Vaccine should be given it again in order to reach sufficient anti-HBs levels.%目的观察乙型肝炎基因重组疫苗的安全性和对大学生的免疫效果,为制定免疫策略提供依据。方法应用乙肝酵母重组疫苗,按0,1和2月3针接种乙肝血清标志物全阴的156名大学生,并用定量放免法(PIA)对免疫后的抗体反应进行检测。结果未发现接种对象出现严重副反应。完成全程免疫后2个月时,抗体阳转率达81.41%,但大多数接种者抗体滴度(mean geometric titer,GMT)在(10~99)IU/L之间,为低应答水平,只有1.92%的人GMT在100IU/L以上,男、女生之间抗体阳转率及GMT构成分布差异多无显著性。12名既往疫苗接种抗体无应答者复种疫苗后,有7人抗体转阳。结论乙肝重组疫苗安全性较好,对大学生

  20. Functional analyses of GB virus B p13 protein: Development of a recombinant GB virus B hepatitis virus with a p7 protein

    DEFF Research Database (Denmark)

    Takikawa, Shingo; Engle, Ronald E; Emerson, Suzanne U;

    2006-01-01

    substitutions of the -1 and/or -3 residues of the putative cleavage sites (amino acid 613/614 and 732/733) abolished processing in vitro and rendered an infectious GBV-B clone nonviable in tamarins. Internal cleavage was predicted at two sites (amino acid 669/670 and 681/682), and in vitro analysis indicated...... availability of a recombinant GBV-B virus containing a p7 protein with similarities to the HCV p7 will enhance the relevance of this model and will be of importance for identifying compounds that inhibit p7 function as additional therapeutic agents....

  1. Recombination of IL18-HBV S Gene Vaccine to Resist Tumor and Hepatitis B%抗肿瘤和乙肝双效IL18-HBV S基因疫苗的构建

    Institute of Scientific and Technical Information of China (English)

    何淑雅; 薛志红; 任为; 刘映霞; 尹志华

    2001-01-01

    [Purpose] To find the combination function of 1L-18 and HBV S gene. [Method] 1L-18 cDNA was obtained by RT-PCR from human embryo liver tissue. And an eukaryote expression vector -pEGFP-N1/ IL-18 with a report gene which was expressing green fluorescene protein was constructed. Then the authors got the IL-18 cDNA by restriction enzyme and PCR. Because of its liverphagy and immunization, the authors used HBV S gene from an expression vector-pcDNA3.0/S and linked it with 1L-18 cDNA. [Result]This two genes were subcloned into an expression eukaryote vector-pIRES-EGFP and formed a new expression vector pIRES-EGFP-S-IL18 that would resist tumor and Hepatitis B.[Conclusion]The recombination of IL18-HBV S gene makes great progress in enhancing the immunity of HBV gene vaccine. It could play a basic role in resisted rumor and Hepatitis B.%[目的]探讨IL-18及乙肝病毒s基因(HBV s)的联合功效。[方法]从胎肝组织中抽提总RNA,RT-PCR扩增IL-18cDNA基因。从载体pcDNA3.0/S中限制性酶切获取HBV S基因,然后,将其与IL—18 cDNA一并亚克隆到真核表达质粒pIRES—EGFP中。[结果]构建了具有抗肿瘤及乙肝双重功效的IL18-HBV S乙肝基因疫苗。[结论]含有IL-18基因的HBv基因疫苗的构建为基因疫苗的功能和应用研究提供了基础。

  2. A recombinant nucleocapsid-based indirect ELISA for serodiagnosis of Rift Valley fever in African wildlife

    International Nuclear Information System (INIS)

    An indirect ELISA (I-ELISA) based on the recombinant nucleocapsid protein (rNp) of Rift Valley fever virus (RVFV) was evaluated for the detection of specific serum IgG antibody in African wildlife. Data sets derived from field-collected sera (n = 918) in Africa (antelopes = 570, black rhinoceros = 43, common zebra = 24, elephant = 73, giraffe = 81, grevy zebra = 78, warthog = 49) were categorised according to the results of a virus neutralisation test (VNT). At cut-offs optimised by the two-graph receiver operating characteristics analysis, the diagnostic sensitivity of the I-ELISA was 100% and diagnostic specificity ranged from 99.8% to 100% while estimates for the Youden's index (J) and efficiency (Ef) ranged from 0.99 to 1 and from 99.7% to 100%, respectively. The rNp-based I-ELISA is highly accurate, safe, and offers a single assay format for rapid detection of IgG antibody to RVFV in sera of different wildlife species. (author)

  3. Hepatitis B and Hepatitis C Infections and Hepatitis B Vaccination in Pediatric Patients on Haemodialysis

    OpenAIRE

    Bak, Mustafa; Aksu, Nejat; Caner KABASAKAL; Cura, Alphan

    1993-01-01

    This study was performed to determine the prevalence hepatitis B surfage antigen other hepatitis B markers anti HCV antibodies the variations during the course of haemodialysis and immun response to a recombinant hepatitis B vaccine in children with chronic renal failure on haemodialysis nbsp; 54 patients 26 males and 28 females median age 12 1±2 7 years range 5 16 years treated with haemodialysis for median of 12 9±13 9 months were studied All hepatitis markers were detected in serum by an e...

  4. [Establishment of hepatitis B virus (HBV) chronic infection mouse model by in vivo transduction with a recombinant adeno-associated virus 8 carrying 1. 3 copies of HBV genome (rAAN8-1. 3HBV)].

    Science.gov (United States)

    Dong, Xiao-Yan; Yu, Chi-Jie; Wang, Gang; Tian, Wen-Hong; Lu, Yue; Zhang, Feng-Wei; Wang, Wen; Wang, Yue; Tan, Wen-Jie; Wu, Xiao-Bing

    2010-11-01

    In this report, we developed a HBV infection model in C57BL/6 mouse line by in vivo injection of a recombinant adeno-associated virus 8 vector carrying 1. 3 copies of HBV genome (ayw subtype) (rAAV8-1. 3HBV). We firstly prepared and purified the rAAV8-1. 3HBV and then injected it into three C57BL/6 mice with the dose of 2 x 10e11vg, respectively. HBsAg and HBeAg were assayed in sera collected at different time points post injection. Ten weeks post injection, the three mice were sacrificed and blood and liver tissue were taken for assay. Copies of HBV DNA were detected by real time PCR and the way of HBV DNA replication was identified by PCR. Subsequently, detection of HBV antigen by immunohistochemistry and pathology analysis of liver tissue of mice were performed. The results suggested that expression of HBsAg and HBeAg lasted for at least 10 weeks in mice sera. Among mice injected with rAAV8-1. 3HBV, HBsAg levels were showed an 'increasing-decreasing-increasing' pattern (the lowest level at the 4th week post injection), while HBeAg levels were kept high and relatively stable. HBV DNA copies were 4.2 x 10(3), 3.6 x 10(3), 2.5 x 10(3) copies/mL in sera and 8.0 x 10(6), 5.7 x 10(6), 2.6 x 10(6) copies/g in hepatic tissues of three mice, respectively. We found that the linear 1. 3HBV DNA in the rAAV8-1. 3HBV could self form into circular HBV genome and replicate in livers of HBV transfected mice. HBsAg and HBcAg were both positive in liver tissue of mice injected with rAAV8-1. 3HBV and no obvious pathological characters were found in liver of mice injected with rAAV8-1. 3HBV. In conclusion, we successfully developed a HBV chronic infection model in C57BL/6 mouse line by in vivo transduction with the recombinant virus rAAV8-1. 3HBV, in which HBV genes could be continuously expressed and replicated over 10 weeks, and paved a way for further characterization of the human chronic hepatitis B virus infection and evaluation of vaccine and anti-HBV agents. PMID:21344744

  5. Interface Recombination in Depleted Heterojunction Photovoltaics based on Colloidal Quantum Dots

    KAUST Repository

    Kemp, Kyle W.

    2013-03-26

    Interface recombination was studied in colloidal quantum dot photovoltaics. Optimization of the TiO2 -PbS interface culminated in the introduction of a thin ZnO buffer layer deposited with atomic layer deposition. Transient photovoltage measurements indicated a nearly two-fold decrease in the recombination rate around 1 sun operating conditions. Improvement to the recombination rate led to a device architecture with superior open circuit voltage (VOC) and photocurrent extraction. Overall a 10% improvement in device efficiency was achieved with Voc enhancements up to 50 mV being realized. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Immune therapy including dendritic cell based therapy in chronic hepatitis B virus infection

    Institute of Scientific and Technical Information of China (English)

    Sk Md Fazle Akbar; Norio Horiike; Morikazu Onji

    2006-01-01

    Hepatitis B virus (HBV) infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of chronic HBV carriers suffer from progressive liver diseases. In addition, all HBV carriers are permanent source of this virus. There is no curative therapy for chronic HBV carriers. Antiviral drugs are recommended for about 10% patients, however, these drugs are costly, have limited efficacy, and possess considerable side effects.Recent studies have shown that immune responses of the host to the HBV are critically involved at every stage of chronic HBV infection: (1) These influence acquisition of chronic HBV carrier state, (2) They are important in the context of liver damages, (3) Recovery from chronic HBV-related liver diseases is dependent on nature and extent of HBV-specific immune responses.However, induction of adequate levels of HBV-specific immune responses in chronic HBV carriers is difficult.During the last one decade, hepatitis B vaccine has been administered to chronic HBV carriers as a therapeutic approach (vaccine therapy). The present regimen of vaccine therapy is safe and cheap, but not so effective.A dendritic cell-based therapeutic vaccine has recently been developed for treating chronic HBV infection. In this review, we will discuss about the concept, scientific logics, strategies and techniques of development of HBV-specific immune therapies including vaccine therapy and dendritic cell-based vaccine therapy for treating chronic HBV infection.

  7. [A primary study of evolution of hepatitis B virus based on motif discovery].

    Science.gov (United States)

    Ma, Lei; Yi, Qing-Qing; Zhang, Qi; He, Jian-Feng

    2014-01-01

    Hepatitis B is a serious infectious disease worldwide, and hepatitis B virus (HBV) is the direct cause of this disease. In recent years, as an essential part of its evolutionary process, HBV mutation has been extensively studied domestically and globally. However, the study on the conserved sequences in HBV sequences is still in its infancy. In this study, we applied multiple EM for motif elicitation (MEME) algorithm to discover HBV motif and proposed a new metric, conservative index (CI), to carry out phylogenetic analysis based on HBV sequences. Then, the constructed phylogenetic tree was subjected to reliability assessment. The results demonstrated that the new metric CI combined with the MEME algorithm can effectively help to discover motifs in HBV sequences and construct a phylogenetic tree based on them and to analyze the evolutionary relationship between HBV sequences; in addition, the possible ancestral sequences of samples may be obtained by conservative analysis. The proposed method is valuable for the exploratory study on large HBV sequence data sets. PMID:24772892

  8. A Follow-up on Efficacy of Immunization of Yeast-derived Recombinant Hepatitis B Vaccine after 11 Years%新生儿重组酵母乙型肝炎疫苗免疫11年效果随访

    Institute of Scientific and Technical Information of China (English)

    吴维寿; 孙超美; 姜铭波; 张国华; 胡凯; 牟文; 宿飞; 周子荣

    2011-01-01

    目的 评价新生儿接种国产重组(酵母)乙型肝炎疫苗后11年的免疫效果.方法 对上海市黄浦区1997年出生并接种重组(酵母)乙型肝炎疫苗的新生儿于免疫后11年进行随访,采血检测乙型肝炎病毒表面抗原(HBsAg),乙型肝炎病毒表面抗体(抗-HBs)和乙型肝炎病毒核心抗体(抗-HBc),1998年开始对乙型肝炎免疫人群开展急性乙型肝炎发病监测.结果 随访检测HBsAg阳性率为0.54%,较免疫前本底的HBsAg阳性率呈较大幅度下降,疫苗保护率为88.05%(95%可信区间为85.17% ~91.17%).接受重组(酵母)乙型肝炎疫苗免疫的对象中,无一例急性乙型肝炎病例报告.结论重组(酵母)乙型肝炎疫苗有较好的远期保护效果,免疫人群尚无再免疫的需要.%[Objective] To evaluate the immune efficacy after 11 years' Immunization of domestic yeast-derived recombinant hepatitis B vaccine in newboms. [Methods] A follow-up on children who were born in 1997 and vaccinated with domestic yeast-derived re-combinant hepatitis B vaccine at their birth was implemented to test the HBsAg, anti-HBs and anti-HBc. The incidence of acute hep-atitis B among subjects with hepatitis B vaccinated was monitored since 1998. [ Results] The positive rate of HBsAg was 0. 54% , compared with that before vaccination in the same area, the positive rate of HBsAg showed a substantially decrease, the vaccine pro-tective efficacy was 88.05% (95% confidence intervals 85. 17% ~91. 17%). Acute hepatitis B was not found in children immu-nized with yeast-derived recombinant hepatitis B vaccine. [ Conclusion ] The domestic yeast recombinant hepatitis B vaccine has a long-term protective effect, and a booster dose is not necessary currently.

  9. Segmentation of hepatic vessels from MRI images for planning of electroporation-based treatments in the liver

    Directory of Open Access Journals (Sweden)

    Marcan Marija

    2014-09-01

    Full Text Available Introduction. Electroporation-based treatments rely on increasing the permeability of the cell membrane by high voltage electric pulses delivered to tissue via electrodes. To ensure that the whole tumor is covered by the sufficiently high electric field, accurate numerical models are built based on individual patient geometry. For the purpose of reconstruction of hepatic vessels from MRI images we searched for an optimal segmentation method that would meet the following initial criteria: identify major hepatic vessels, be robust and work with minimal user input.

  10. A universal cloning method based on yeast homologous recombination that is simple, efficient, and versatile

    OpenAIRE

    Joska, Tammy M.; Mashruwala, Ameya; Boyd, Jeffrey M.; Belden, William J.

    2014-01-01

    Cloning by homologous recombination (HR) in Saccharomyces cerevisiae is an extremely efficient and cost-effective alternative to other methods of recombinant DNA technologies. Unfortunately, it is incompatible with all the various specialized plasmids currently used in microbiology and biomedical research laboratories, and is therefore, not widely adopted. In an effort to dramatically improve the versatility of yeast gap-repair cloning and make it compatible with any DNA plasmid, we demonstra...

  11. Nanog reporter system in mouse embryonic stem cells based on highly efficient BAC homologous recombination

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Nanog is a novel transcription factor specifically expressed in mouse embryonic stem cells (mES cells). It has been reported that Nanog plays an essential role in maintaining multi-potency of ES cells. The expression of Nanog is very sensitive to ES cells differentiation, making Nanog one of the best markers to indicate the status of ES cells. In this study, we developed an efficient method to construct Nanog promoter driven EGFP reporter system based on the BAC homologous recombination. We further generated a Nanog-EGFP reporter mES cell line. This reporter mES cell line exhibited features similar to those of normal mES cells, and the EGFP reporter efficiently reflected the expression of Nanog, indicating the differentiation status of mES cells. We achieved a reliable experimental reporter system to research self-renewal and differentiation of mES cells. The system could facilitate research on culture system of mES cells and researches on the expression and regulation of Nanog and other related factors in mES cells.

  12. Rapid and Robust PCR-Based All-Recombinant Cloning Methodology.

    Science.gov (United States)

    Dubey, Abhishek Anil; Singh, Manika Indrajit; Jain, Vikas

    2016-01-01

    We report here a PCR-based cloning methodology that requires no post-PCR modifications such as restriction digestion and phosphorylation of the amplified DNA. The advantage of the present method is that it yields only recombinant clones thus eliminating the need for screening. Two DNA amplification reactions by PCR are performed wherein the first reaction amplifies the gene of interest from a source template, and the second reaction fuses it with the designed expression vector fragments. These vector fragments carry the essential elements that are required for the fusion product selection. The entire process can be completed in less than 8 hours. Furthermore, ligation of the amplified DNA by a DNA ligase is not required before transformation, although the procedure yields more number of colonies upon transformation if ligation is carried out. As a proof-of-concept, we show the cloning and expression of GFP, adh, and rho genes. Using GFP production as an example, we further demonstrate that the E. coli T7 express strain can directly be used in our methodology for the protein expression immediately after PCR. The expressed protein is without or with 6xHistidine tag at either terminus, depending upon the chosen vector fragments. We believe that our method will find tremendous use in molecular and structural biology. PMID:27007922

  13. Vaccine delivery system for tuberculosis based on nano-sized hepatitis B virus core protein particles

    Directory of Open Access Journals (Sweden)

    Dhanasooraj D

    2013-02-01

    Full Text Available Dhananjayan Dhanasooraj, R Ajay Kumar, Sathish MundayoorMycobacterium Research Group, Rajiv Gandhi Centre for Biotechnology, Kerala, IndiaAbstract: Nano-sized hepatitis B virus core virus-like particles (HBc-VLP are suitable for uptake by antigen-presenting cells. Mycobacterium tuberculosis antigen culture filtrate protein 10 (CFP-10 is an important vaccine candidate against tuberculosis. The purified antigen shows low immune response without adjuvant and tends to have low protective efficacy. The present study is based on the assumption that expression of these proteins on HBc nanoparticles would provide higher protection when compared to the native antigen alone. The cfp-10 gene was expressed as a fusion on the major immunodominant region of HBc-VLP, and the immune response in Balb/c mice was studied and compared to pure proteins, a mixture of antigens, and fusion protein-VLP, all without using any adjuvant. The humoral, cytokine, and splenocyte cell proliferation responses suggested that the HBc-VLP bearing CFP-10 generated an antigen-specific immune response in a Th1-dependent manner. By virtue of its self-adjuvant nature and ability to form nano-sized particles, HBc-VLPs are an excellent vaccine delivery system for use with subunit protein antigens identified in the course of recent vaccine research.Keywords: Mycobacterium tuberculosis, VLP, hepatitis B virus core particle, CFP-10, self-adjuvant, vaccine delivery

  14. Children's yeast recombinant hepatitis b vaccine immunization effect assessment%儿童接种重组酵母乙型肝炎疫苗的免疫效果评价

    Institute of Scientific and Technical Information of China (English)

    唐林锋

    2015-01-01

    目的:评价儿童接种重组酵母乙型肝炎疫苗的免疫效果。方法:采用随机整群抽样法选取489例接种重组酵母乙型肝炎疫苗儿童,接种后检测血清HBsAg、抗-HBs和抗-HBc水平,随访观察免疫效果。结果:本组489例儿童在免疫接种1年后和5年后的HBsAg阳性率、抗-H B c阳性率之间的差异无统计学意义(P>0.05),免疫接种5年后的抗-H B s阳性率为33.95%,明显低于免疫接种1年后的93.56%,差异有统计学意义(P 0.05), after five years in the immunization of anti - HBs positive rate was 33.95%, significantly less than 1 year after immunization of 93.56%, the difference was statistically significant (P < 0.05).Conclusions: Children inoculated yeast recombinant hepatitis b vaccine can achieve lasting and effective immune effect, worthy of further promotion of use.

  15. Analysis on Immune Effect and Safety of Recombinant Hepatitis B Vaccine in Healthy Population%重组乙型肝炎疫苗在健康人群中的免疫效果及安全性评价

    Institute of Scientific and Technical Information of China (English)

    钟群; 谌稳国; 罗述斌

    2012-01-01

    目的 分析常用的两种国产重组乙肝疫苗在健康人群中接种的免疫效果及安全性. 方法 选择2010年1月-2011年1月在本门诊接种乙肝疫苗的532例健康人群,按接种疫苗类别分为A组(酵母疫苗10 μg)与B组(CHO疫苗20μg),观察两组间的不良反应发生率和发生程度.于第2、3针接种前及3针全程接种后1月和1年时分别应用放射免疫法(RIA)检测血清HBsAb滴度水平,并比较两组抗体的阳性率. 结果 两组疫苗接种不良反应发生率差异无统计学意义(P>0.05).全程接种后1月时,B组的HBsAb GMT峰值高于A组(P<0.05).全程接种后1年B组抗体阳性率和HBsAb滴度均高于A组(P<0.05). 结论 在可按受的、等同少而轻的接种不良反应基础上,应用20 μg的CHO疫苗可使健康人群获得更大的保护效力.%Objective To analyze the immune effect and safety of two kinds of homemade recombinant hepatitis B vaccines in healthy people. Methods Altogether 532 cases of healthy people with hepatitis R vaccines in Outpatient Department from January 2010 to January 2011 were divided into group A (yeast vaccine, lOjug) and group B (CHO vaccine, 20jng) according to the category of vaccines. The incidence and degree of adverse reactions were observed between the two groups. The serum HBsAb titers were detected with radioimmunoassay before the second and third shots, one month and one year after the full course vaccination. The antibody - positive rate was compared between the two groups. Results There was no statistically significant difference in the incidence of adverse reaction between the two groups (F>0. 05). One month after the full course vaccination, the HBsAb GMT peak value of group B was significantly higher than that in group A (P< 0.05). One year after the full course vaccination, the antibody - positive rate and HBsAb titers in group B were both significantly higher than those in group A (P<0.05). Conclusions On the basis of acceptable

  16. Novel vaccination approach for dengue infection based on recombinant immune complex universal platform.

    Science.gov (United States)

    Kim, Mi-Young; Reljic, Rajko; Kilbourne, Jacquelyn; Ceballos-Olvera, Ivonne; Yang, Moon-Sik; Reyes-del Valle, Jorge; Mason, Hugh S

    2015-04-01

    Dengue infection is on the rise in many endemic areas of the tropics. Vaccination remains the most realistic strategy for prevention of this potentially fatal viral disease but there is currently no effective vaccine that could protect against all four known serotypes of the dengue virus. This study describes the generation and testing of a novel vaccination approach against dengue based on recombinant immune complexes (RIC). We modelled the dengue RIC on the existing Ebola RIC (Phoolcharoen, et al. Proc Natl Acad Sci USA 2011;108(Dec (51)):20695) but with a key modification that allowed formation of a universal RIC platform that can be easily adapted for use for other pathogens. This was achieved by retaining only the binding epitope of the 6D8 ant-Ebola mAb, which was then fused to the consensus dengue E3 domain (cEDIII), resulting in a hybrid dengue-Ebola RIC (DERIC). We expressed human and mouse versions of these molecules in tobacco plants using a geminivirus-based expression system. Following purification from the plant extracts by protein G affinity chromatography, DERIC bound to C1q component of complement, thus confirming functionality. Importantly, following immunization of mice, DERIC induced a potent, virus-neutralizing anti-cEDIII humoral immune response without exogenous adjuvants. We conclude that these self-adjuvanting immunogens have the potential to be developed as a novel vaccine candidate for dengue infection, and provide the basis for a universal RIC platform for use with other antigens. PMID:25728317

  17. Association between chronic viral hepatitis infection and breast cancer risk: a nationwide population-based case-control study

    Directory of Open Access Journals (Sweden)

    Su Fu-Hsiung

    2011-11-01

    Full Text Available Abstract Background In Taiwan, there is a high incidence of breast cancer and a high prevalence of viral hepatitis. In this case-control study, we used a population-based insurance dataset to evaluate whether breast cancer in women is associated with chronic viral hepatitis infection. Methods From the claims data, we identified 1,958 patients with newly diagnosed breast cancer during the period 2000-2008. A randomly selected, age-matched cohort of 7,832 subjects without cancer was selected for comparison. Multivariable logistic regression models were constructed to calculate odds ratios of breast cancer associated with viral hepatitis after adjustment for age, residential area, occupation, urbanization, and income. The age-specific ( Results There were no significant differences in the prevalence of hepatitis C virus (HCV infection, hepatitis B virus (HBV, or the prevalence of combined HBC/HBV infection between breast cancer patients and control subjects (p = 0.48. Multivariable logistic regression analysis, however, revealed that age Conclusions HCV infection, but not HBV infection, appears to be associated with early onset risk of breast cancer in areas endemic for HCV and HBV. This finding needs to be replicated in further studies.

  18. Base composition, selection, and phylogenetic significance of indels in the recombination activating gene-1 in vertebrates

    Directory of Open Access Journals (Sweden)

    Vences Miguel

    2009-12-01

    Full Text Available Abstract Background The Recombination Activating Proteins, RAG1 and RAG2, play a crucial role in the immune response in vertebrates. Among the nuclear markers currently used for phylogenetic purposes, Rag1 has especially enjoyed enormous popularity, since it successfully contributed to elucidating the relationships among and within a large variety of vertebrate lineages. We here report on a comparative investigation of the genetic variation, base composition, presence of indels, and selection in Rag1 in different vertebrate lineages (Actinopterygii, Amphibia, Aves, Chondrichthyes, Crocodylia, Lepidosauria, Mammalia, and Testudines through the analysis of 582 sequences obtained from Genbank. We also analyze possible differences between distinct parts of the gene with different type of protein functions. Results In the vertebrate lineages studied, Rag1 is over 3 kb long. We observed a high level of heterogeneity in base composition at the 3rd codon position in some of the studied vertebrate lineages and in some specific taxa. This result is also paralleled by taxonomic differences in the GC content at the same codon position. Moreover, positive selection occurs at some sites in Aves, Lepidosauria and Testudines. Indels, which are often used as phylogenetic characters, are more informative across vertebrates in the 5' than in the 3'-end of the gene. When the entire gene is considered, the use of indels as phylogenetic character only recovers one major vertebrate clade, the Actinopterygii. However, in numerous cases insertions or deletions are specific to a monophyletic group. Conclusions Rag1 is a phylogenetic marker of undoubted quality. Our study points to the need of carrying out a preliminary investigation on the base composition and the possible existence of sites under selection of this gene within the groups studied to avoid misleading resolution. The gene shows highly heterogeneous base composition, which affects some taxa in particular and

  19. Evaluation of factors associated with relapse in telaprevir-based triple therapy for chronic hepatitis C

    Science.gov (United States)

    Kondo, C; Atsukawa, M; Tsubota, A; Shimada, N; Abe, H; Aizawa, Y

    2016-01-01

    Background and Rationale: Most patients with chronic hepatitis C show virological response to telaprevir-based triple therapy, and achieve an end-of-treatment response (ETR). However, some patients showing ETR develop virological relapse. This study was carried out to evaluate factors associated with relapse after triple therapy. Materials and Methods: A prospective, multicentric study was conducted in chronic hepatitis C patients who received telaprevir-based triple therapy. We evaluated independent variables such as age, with or without cirrhosis, prior treatment response to interferon (IFN) therapy, IL28B genotype, core amino acid (aa) 70 mutation, drug adherence, white blood cell counts, hemoglobin level, and serum low-density lipoprotein (LDL) cholesterol level. The characteristics of the patients who relapsed after achieving ETR were compared with those who did not. Results: Among 168 patients, 157 patients achieved ETR (93.5%) and 11 discontinued. Of these 157 patients, relapse occurred in 21 patients (13.4%). Nineteen patients (90.5%) of 21 relapsed patients had the IL28B non-TT genotype (P = 1.79 × 10-9). Multivariate analysis identified core amino acid 70 [P = 0.018, crude odds ratio (OR): 6.927] and the IL28B genotype (P = 3.758 × 10-5, crude OR: 39.311) as significantly independent factors that influenced the relapse-related variables. Among the 49 patients with the IL28B non-TT, 18 patients had core aa70 mutation and 31 patients had core aa70 wild-type. In addition, 66.7% (12/18) of those with core aa70 mutation and 22.6% (7/31) of those with core aa70 wild-type developed relapse (P = 0.005). Discussion: Core aa70 mutation and the IL28B non-TT genotype were identified as independent factors that influenced relapse after achievement of ETR for telaprevir-based triple therapy. PMID:26732192

  20. A Vesicular Stomatitis Virus-Based Therapeutic Vaccine Generates a Functional CD8 T Cell Response to Hepatitis B Virus in Transgenic Mice

    OpenAIRE

    Cobleigh, Melissa A.; Wei, Xin; Robek, Michael D.

    2013-01-01

    Recombinant vesicular stomatitis virus (VSV) is a promising therapeutic vaccine platform. Using a transgenic mouse model of chronic hepatitis B virus (HBV) infection, we evaluated the therapeutic potential of a VSV vector expressing the HBV middle surface envelope glycoprotein (MS). VSV-MS immunization generated HBV-specific CD8 T cell and antibody responses in transgenic mice that express low HBV antigen levels. These findings support the further development of VSV as a therapeutic vaccine v...

  1. Recombinant and epitope-based vaccines on the road to the market and implications for vaccine design and production.

    Science.gov (United States)

    Oyarzún, Patricio; Kobe, Bostjan

    2016-03-01

    Novel vaccination approaches based on rational design of B- and T-cell epitopes - epitope-based vaccines - are making progress in the clinical trial pipeline. The epitope-focused recombinant protein-based malaria vaccine (termed RTS,S) is a next-generation approach that successfully reached phase-III trials, and will potentially become the first commercial vaccine against a human parasitic disease. Progress made on methods such as recombinant DNA technology, advanced cell-culture techniques, immunoinformatics and rational design of immunogens are driving the development of these novel concepts. Synthetic recombinant proteins comprising both B- and T-cell epitopes can be efficiently produced through modern biotechnology and bioprocessing methods, and can enable the induction of large repertoires of immune specificities. In particular, the inclusion of appropriate CD4+ T-cell epitopes is increasingly considered a key vaccine component to elicit robust immune responses, as suggested by results coming from HIV-1 clinical trials. In silico strategies for vaccine design are under active development to address genetic variation in pathogens and several broadly protective "universal" influenza and HIV-1 vaccines are currently at different stages of clinical trials. Other methods focus on improving population coverage in target populations by rationally considering specificity and prevalence of the HLA proteins, though a proof-of-concept in humans has not been demonstrated yet. Overall, we expect immunoinformatics and bioprocessing methods to become a central part of the next-generation epitope-based vaccine development and production process. PMID:26430814

  2. Recombinant and epitope-based vaccines on the road to the market and implications for vaccine design and production.

    Science.gov (United States)

    Oyarzún, Patricio; Kobe, Bostjan

    2016-03-01

    Novel vaccination approaches based on rational design of B- and T-cell epitopes - epitope-based vaccines - are making progress in the clinical trial pipeline. The epitope-focused recombinant protein-based malaria vaccine (termed RTS,S) is a next-generation approach that successfully reached phase-III trials, and will potentially become the first commercial vaccine against a human parasitic disease. Progress made on methods such as recombinant DNA technology, advanced cell-culture techniques, immunoinformatics and rational design of immunogens are driving the development of these novel concepts. Synthetic recombinant proteins comprising both B- and T-cell epitopes can be efficiently produced through modern biotechnology and bioprocessing methods, and can enable the induction of large repertoires of immune specificities. In particular, the inclusion of appropriate CD4+ T-cell epitopes is increasingly considered a key vaccine component to elicit robust immune responses, as suggested by results coming from HIV-1 clinical trials. In silico strategies for vaccine design are under active development to address genetic variation in pathogens and several broadly protective "universal" influenza and HIV-1 vaccines are currently at different stages of clinical trials. Other methods focus on improving population coverage in target populations by rationally considering specificity and prevalence of the HLA proteins, though a proof-of-concept in humans has not been demonstrated yet. Overall, we expect immunoinformatics and bioprocessing methods to become a central part of the next-generation epitope-based vaccine development and production process.

  3. Hepatitis Vaccines

    OpenAIRE

    Ogholikhan, Sina; Schwarz, Kathleen B

    2016-01-01

    Viral hepatitis is a serious health problem all over the world. However, the reduction of the morbidity and mortality due to vaccinations against hepatitis A and hepatitis B has been a major component in the overall reduction in vaccine preventable diseases. We will discuss the epidemiology, vaccine development, and post-vaccination effects of the hepatitis A and B virus. In addition, we discuss attempts to provide hepatitis D vaccine for the 350 million individuals infected with hepatitis B ...

  4. Digital medical technology based on 64-slice computed tomography in hepatic surgery

    Institute of Scientific and Technical Information of China (English)

    FANG Chi-hua; HUANG Yan-peng; CHEN Mian-ling; LU Chao-min; LI Xiao-feng; QIU Wen-feng

    2010-01-01

    Background With the rapid development of computer technology, digital medicine has become a new direction in surgery. The application of digital medicine in hepatic surgery is still at the early stage and less reported in the literature. The aim of this study was to apply digital medical technology in the context of hepatic surgery. Methods Data from 64-slice helical computed tomography of 17 patients, including 13 with hepatocellular carcinoma and 4 with hepatic hemangioma, were imported into independently developed medical image software program, segmentation and three-dimensional reconstruction were performed. The three-dimensional models were then processed with the FreeForm Modeling System. We used virtual surgical instruments to perform surgery on the models. Simulated surgeries included six hepatic segmentectomies, four left hemihepatectomies, three right hemihepatectomies for hepatocellular carcinoma, one hepatic segmentectomy, two stripping surgeries, and one irregular segmentectomy combined with stripping surgery for hemangioma. For resections involving more than three hepatic segments, total and residual functional hepatic volumes were measured before and after simulation surgery, and the resection ratio was calculated.Results The anatomy of the models was distinct and was used to localize lesions. We used virtual surgical instruments to perform simulated surgeries and used the models to optimize actual surgeries. We were able to minimize resection volume as well as surgical risk.Conclusions Digital medical technology is helpful in the diagnosis of hepatic disease and in optimizing surgical plans. Three-dimensional models can decrease surgical risk and help prevent postoperative hepatic failure.

  5. Magnetic monitoring of the hydrogenation-decomposition-desorption-recombination process in SmFe-based alloys

    International Nuclear Information System (INIS)

    A clear understanding of the reactions and phase changes that occur during the processing of a material is vital if we want to be able to achieve the best properties. In this series of experiments, we have used a vibrating-sample magnetometer (VSM) to monitor the magnetic state of compacted powder samples of Sm13.7Fe86.3 and Sm13.8Fe82.2Ta4.0 during a standard hydrogenation-disproportionation-desorption-recombination (HDDR) cycle. The samples were mounted in the VSM and heated at 4 deg.C/min in 1 bar of flowing hydrogen to 750 deg.C, they were then held for 60 min under hydrogen and approximately 120 min under vacuum before being cooled to room temperature under vacuum at 4 deg.C/min. The results show that the initial absorption of hydrogen by the Sm2Fe17-type phase results in an increase in magnetization as well as TC. At higher temperatures, the disproportionation reactions of the Sm13.7Fe86.3 and Sm13.8Fe82.2Ta4.0 materials were observed to proceed at a faster rate for the second and third cycles of the HDDR procedure than for the first. The results are in agreement with our previous transmission electron microscopy studies, which showed that much of the Ta that was initially dissolved in the Sm2Fe17-type phase of the cast Sm13.8Fe82.2Ta4.0 material was reformed as Ta-based precipitates after a single HDDR cycle leaving the Sm2Fe17-type phase with much less dissolved Ta

  6. Development of Recombinant Nucleoprotein-Based Diagnostic Systems for Lassa Fever▿

    OpenAIRE

    Saijo, Masayuki; Georges-Courbot, Marie-Claude; Marianneau, Philippe; Romanowski, Victor; Fukushi, Shuetsu; Mizutani, Tetsuya; Georges, Alain-Jean; Kurata, Takeshi; Kurane, Ichiro; Morikawa, Shigeru

    2007-01-01

    Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa ce...

  7. Model-based adaptive control of acetate concentration during the production of recombinant proteins with E. coli

    OpenAIRE

    Rocha, I; Ferreira, E. C.

    2003-01-01

    A model-based adaptive linearizing control law was derived for the regulation of the acetate concentration during the fed-batch fermentation of recombinant proteins with high cell density culture of Escherichia coli growing on glucose. An unstructured model for the growth was applied to the major metabolic pathways: oxidative growth on glucose, fermentative growth on glucose, oxidative growth on acetate, and maintenance. A model order reduction method was used to allow the d...

  8. Interruption of perinatal transmission of hepatitis B virus(HBV) with a recombinant yeast derived hepatitis B vaccine%国产重组酵母乙型肝炎疫苗阻断乙型肝炎母婴传播的研究

    Institute of Scientific and Technical Information of China (English)

    李艳萍; 李荣成; 杨进业; 李坚龙; 徐桂生; 李琼池; 黎国形; 胡可能

    2001-01-01

    目的探讨乙型肝炎基因工程疫苗阻断 HBV母婴传播的免疫保护效果和免疫策略。方法对 HBsAg和 HBeAg 同时阳性母亲的 169例新生儿接种国产重组酵母乙型肝炎疫苗,于免后 3、 9、 12、 24、 36、 48月采血进行血清免疫学追踪观察。结果 22例婴儿 1岁前 HBsAg阳性, 19例( 11.24%)成为慢性携带者,免后一年阻断保护率为 85.94%。抗 -HBs阳性率和抗 -HBs滴度均于 9~ 12月龄时达一高峰, 24和 36月龄时有所下降,免后 1~ 4年抗 -HBs阳性率分别为 96.43%、 91.07%、 85.19%和 70.00%。结论国产酵母重组乙型肝炎疫苗对 HBV母婴传播具有良好免疫阻断效果。母亲 HBsAg和 HBeAg双阳性的幼儿在 3~ 4岁时需加强免疫。%Objective To assess the efficacy and immunogenicity of recombinant yeast derived hepatitis B vaccine in infants born to HBsAg and HBeAg carrier mother.Methods A total of 169 neonates born to HBsAg,HBeAg both positive mothers were vaccinated with 5,5,5 μ g doses of recombinant yeast derived hepatitis B vaccine by 0,1,6 months schedule.They were all followed for 3,9,12,24,36,48 months after the primary vaccination.Results 19 infants (11.24%) become HBsAg positively conversed in one year after primary vaccination,and the protective efficacy rate of interruption of perinatal transmission was 85.94%.The peak of conversion rate and titer of anti-HBs were observed in aged 9~12 months,and it decreased progressively at aged 24~36 months,the positive rates of anti-HBs were 96.43%,91.07%,85.19% and 70.0% ,respectively.Conclusion The recombinant yeast derived hepatitis B vaccine has good immunogenicity and high protective efficacy in interruption of perinatal transmission,a booster dose seems necessary in aged 3-4 years after vaccination.

  9. Differentiation of pyogenic hepatic abscesses from malignant mimickers using multislice-based texture acquired from contrast-enhanced computed tomography

    Institute of Scientific and Technical Information of China (English)

    Shi-Teng Suo; Zhi-Guo Zhuang; Meng-Qiu Cao; Li-Jun Qian; Xin Wang; Run-Lin Gao; Yu Fan; Jian-Rong Xu

    2016-01-01

    BACKGROUND: Pyogenic hepatic abscess may mimic prima-ry or secondary carcinoma of the liver on contrast-enhanced computed tomography (CECT). The present study was to ex-plore the usefulness of the analysis of multislice-based texture acquired from CECT in the differentiation between pyogenic hepatic abscesses and malignant mimickers. METHODS: This retrospective study included 25 abscesses in 20 patients and 33 tumors in 26 subjects who underwent CECT. To make comparison, we also enrolled 19 patients with hepatic single simple cyst. The images from CECT were ana-lyzed using a Laplacian of Gaussian band-pass iflter (5 iflter levels with sigma weighting ranging from 1.0 to 2.5). We also quantiifed the uniformity, entropy, kurtosis and skewness of the multislice-based texture at different sigma weightings. Sta-tistical signiifcance for these parameters was tested with one-way ANOVA followed by Tukey honestly signiifcant difference (HSD) test. Diagnostic performance was evaluated using the receiver operating characteristic (ROC) curve analysis. RESULTS: There were signiifcant differences in entropy and uniformity at all sigma weightings (P CONCLUSION: Multislice-based texture analysis may be use-ful for differentiating pyogenic hepatic abscesses from malig-nant mimickers.

  10. Dendritic cell-based vaccination with lentiviral vectors encoding ubiquitinated hepatitis B core antigen enhances hepatitis B virus-specific immune responses in vivo.

    Science.gov (United States)

    Dai, Shenglan; Zhuo, Meng; Song, Linlin; Chen, Xiaohua; Yu, Yongsheng; Tang, Zhenghao; Zang, Guoqing

    2015-11-01

    The activity of hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) plays a predominant role in the clearance of HBV. Dendritic cells (DCs) are key antigen-presenting cells and play an important role in the initiation of immune responses. We previously verified that lentiviral vector encoding ubiquitinated hepatitis B core antigen (LV-Ub-HBcAg) effectively transduced DCs to induce maturation, and the mature DCs efficiently induced T cell polarization to Th1 and generated HBcAg-specific CTLs ex vivo. In this study, HBV-specific immune responses of LV-Ub-HBcAg in BALB/c mice (H-2Kd) were evaluated. It was shown that direct injection of LV-Ub-HBcAg increased the production of cytokines IL-2 and IFN-γ, elicited strong antibody responses, and remarkably generated a high percentage of IFN-γ+CD8+ T cells with HBV-specific CTL responses in BALB/c mice. In addition, direct injection of LV-Ub-HBcAg induced potent anti-HBV immune responses, similar to those elicited by in vitro-transduced DCs. In conclusion, the DC-based therapeutic vaccine LV-Ub-HBcAg elicited specific antibody immune responses and induced robust specific CTL activity in vivo. PMID:26373843

  11. Base composition, selection, and phylogenetic significance of indels in the recombination activating gene-1 in vertebrates

    NARCIS (Netherlands)

    Chiari, Y.; Meijden, van der A.; Madsen, O.; Vences, M.; Meyer, A.

    2009-01-01

    Background: The Recombination Activating Proteins, RAG1 and RAG2, play a crucial role in the immune response in vertebrates. Among the nuclear markers currently used for phylogenetic purposes, Rag1 has especially enjoyed enormous popularity, since it successfully contributed to elucidating the relat

  12. Protein biomarker-based screening for detection of recombinant bovine somatotropin abuse in dairy cows

    NARCIS (Netherlands)

    Ludwig, S.K.J.

    2014-01-01

    Recombinant bovine somatotropin (rbST) is a 22 kDa proteohormone, which can be used to increase milk production in dairy cows. It has been marketed since 1994 and while its use in food production is approved in several countries, such as the US, it is banned in the EU since 2000. To enforce the ban

  13. Neuropsychiatric and other side effects of peginterferon-based therapy of chronic hepatitis C infection

    NARCIS (Netherlands)

    G. Bezemer (Geert)

    2012-01-01

    textabstractThe Hepatitis C virus (HCV) was identified in 1989, after extensive research, as a cause of the so-called non-A non-B hepatitis. HCV is an RNA virus belonging to the genus Hepacivirus in the family of Flaviviridae. Six major different genotypes of the virus are discerned. HCV is transmi

  14. Prospective study of differential diagnosis of hepatic tumors by pattern-based classification of contrast-enhanced sonography

    Institute of Scientific and Technical Information of China (English)

    Kazushi Numata; Tetsuo Isozaki; Manabu Morimoto; Kazuya Sugimori; Reiko Kunisaki; Toshio Morizane; Katsuaki Tanaka

    2006-01-01

    AIM: To prospectively evaluate the usefulness of a pattern-based classification of contrast-enhanced sonographic findings for differential diagnosis of hepatic tumors.METHODS: We evaluated the enhancement pattern of the contrast-enhanced sonography images in 586 patients with 586 hepatic lesions, consisting of 383 hepatocellular carcinomas, 89 metastases, and 114 hemangiomas. After injecting a galactose-palmitic acid contrast agent, lesions were scanned by contrastenhanced harmonic gray-scale sonography in three phases: arterial, portal, and late. The enhancement patterns of the initial 303 lesions were classified retrospectively, and multiple logistic regression analysis was used to identify enhancement patterns that allowed differentiation between hepatic tumors. We then used the pattern-based classification of enhancement we had retrospectively devised to prospectively diagnose 283 liver tumors.RESULTS: Seven enhancement patterns were found to be significant predictors of different hepatic tumors.The presence of homogeneous or heterogeneous enhancement both in the arterial and portal phase was the typical enhancement pattern for hepatocellular carcinoma, while the presence of peritumoral vessels in the arterial phase and ring enhancement or a perfusion defect in the portal phase was the typical enhancement pattern for metastases, and the presence of peripheral nodular enhancement both in the arterial and portal phase was the typical enhancement pattern for hemangioma. The sensitivity, specificity, and accuracy of prospective diagnosis based on the combinations of enhancement patterns, respectively, were 93.2%,96.2%, and 94.0% for hepatocellular carcinoma, 87.9%,99.6%, and 98.2% for metastasis, and 95.6%, 94.1%,and 94.3% for hemangioma.CONCLUSION: The pattern-based classification of the contrast-enhanced sonographic findings is useful for differentiating among hepatic tumors.

  15. A Novel Conductometric Urea Biosensor with Improved Analytical Characteristic Based on Recombinant Urease Adsorbed on Nanoparticle of Silicalite.

    Science.gov (United States)

    Velychko, T P; Soldatkin, О О; Melnyk, V G; Marchenko, S V; Kirdeciler, S K; Akata, B; Soldatkin, A P; El'skaya, A V; Dzyadevych, S V

    2016-12-01

    Development of a conductometric biosensor for the urea detection has been reported. It was created using a non-typical method of the recombinant urease immobilization via adsorption on nanoporous particles of silicalite. It should be noted that this biosensor has a number of advantages, such as simple and fast performance, the absence of toxic compounds during biosensor preparation, and high reproducibility (RSD = 5.1 %). The linear range of urea determination by using the biosensor was 0.05-15 mM, and a lower limit of urea detection was 20 μM. The bioselective element was found to be stable for 19 days. The characteristics of recombinant urease-based biomembranes, such as dependence of responses on the protein and ion concentrations, were investigated. It is shown that the developed biosensor can be successfully used for the urea analysis during renal dialysis. PMID:26911570

  16. A Novel Conductometric Urea Biosensor with Improved Analytical Characteristic Based on Recombinant Urease Adsorbed on Nanoparticle of Silicalite

    Science.gov (United States)

    Velychko, T. P.; Soldatkin, O. O.; Melnyk, V. G.; Marchenko, S. V.; Kirdeciler, S. K.; Akata, B.; Soldatkin, A. P.; El'skaya, A. V.; Dzyadevych, S. V.

    2016-02-01

    Development of a conductometric biosensor for the urea detection has been reported. It was created using a non-typical method of the recombinant urease immobilization via adsorption on nanoporous particles of silicalite. It should be noted that this biosensor has a number of advantages, such as simple and fast performance, the absence of toxic compounds during biosensor preparation, and high reproducibility (RSD = 5.1 %). The linear range of urea determination by using the biosensor was 0.05-15 mM, and a lower limit of urea detection was 20 μM. The bioselective element was found to be stable for 19 days. The characteristics of recombinant urease-based biomembranes, such as dependence of responses on the protein and ion concentrations, were investigated. It is shown that the developed biosensor can be successfully used for the urea analysis during renal dialysis.

  17. Laboratory-based surveillance for hepatitis E virus infection, United States, 2005-2012.

    Science.gov (United States)

    Drobeniuc, Jan; Greene-Montfort, Tracy; Le, Ngoc-Thao; Mixson-Hayden, Tonya R; Ganova-Raeva, Lilia; Dong, Chen; Novak, Ryan T; Sharapov, Umid M; Tohme, Rania A; Teshale, Eyasu; Kamili, Saleem; Teo, Chong-Gee

    2013-02-01

    To investigate characteristics of hepatitis E cases in the United States, we tested samples from persons seronegative for acute hepatitis A and B whose clinical specimens were referred to the Centers for Disease Control and Prevention during June 2005-March 2012 for hepatitis E virus (HEV) testing. We found that 26 (17%) of 154 persons tested had hepatitis E. Of these, 15 had not recently traveled abroad (nontravelers), and 11 had (travelers). Compared with travelers, nontravelers were older (median 61 vs. 32 years of age) and more likely to be anicteric (53% vs. 8%); the nontraveler group also had fewer persons of South Asian ethnicity (7% vs. 73%) and more solid-organ transplant recipients (47% vs. 0). HEV genotype 3 was characterized from 8 nontravelers and genotypes 1 or 4 from 4 travelers. Clinicians should consider HEV infection in the differential diagnosis of hepatitis, regardless of patient travel history. PMID:23347695

  18. 转基因植物乙型肝炎疫苗的研究进展%Advances in research of transgenic plant-based hepatitis B vaccines

    Institute of Scientific and Technical Information of China (English)

    袁涛

    2011-01-01

    自Mason等首次报道转基因烟草表达HBsAg以来,可表达抗原的范围和宿主植物的种类得到了快速扩展,HBsAg在多种植物中获得表达.植物合成的HBsAg可形成病毒样颗粒,与酵母及哺乳动物细胞来源的乙型肝炎(乙肝)疫苗具有相似的结构和免疫原性,经注射或口服接种可在动物和人体中诱导免疫应答.口服植物源性乙肝疫苗能诱导体液及黏膜免疫应答,作为黏膜疫苗可以扩大现行注射用重组疫苗保护性应答的范围,是安全、廉价和接种简便的新型候选乙肝疫苗.此文主要介绍乙肝疫苗在转基因植物中的表达及其免疫原性的研究进展.%Since HBsAg was expressed in transgenic tobacco plant and first reported by Mason et al,the range of expressed antigens and the species of plants have been expanded rapidly, and HBsAg has been successfully expressed in various plants. The HBsAg synthesized in plants is able to be assembled into virus-like particles and its molecular structure and immunogenicity are similar to those of subunit HBsAg vaccines produced in yeast or mammalian cells, which has displayed high immunogenicity in animals and human after injection or oral administration. Edible transgenic plant-derived hepatitis B vaccine can induce both humoral and mucosal immune responses,which can be a useful supplement to the currently used recombinant injection vaccines as a mucosal vaccine and expand the range of protective responses. It would be a safe, cheap and easily-inoculated candidate hepatitis B vaccine. This paper reviews the advances in the expression and immunogenicity of plant-based hepatitis B vaccine.

  19. PREVALENCE OF RISK FACTORS FOR HEPATITIS C AND ASSOCIATED FACTORS: a population-based study in southern Brazil

    Directory of Open Access Journals (Sweden)

    David Timm KVITKO

    2013-04-01

    Full Text Available Context The hepatitis C is a severe public health problem worldwide because its consequences. Studies which aim at determining the prevalence of risk factors are really important to understand the problem. Objective To estimate the prevalence and factors associated with some risk factors for the disease in a community, called Restinga, located in the city of Porto Alegre, RS, Brazil. Method This paper is based on a population-based cross-sectional study, with systematic sampling and proportional to the size of census tracts in which 3,391 adults answered a standardized questionnaire. Results The prevalence of blood transfusion among the people who were interviewed was 14.98%, 60.83% of those had it before 1993. A total of 16.16% of the people had a tattoo, 7.23% wore a piercing, 1.09% said they had already injected illicit drugs and 12.39% reported previous hospitalization. Prevalence ratios showed that tattoos were more common among young people, piercings among women and illicit drugs among men. Conclusions To summarize, the recognition of risk factors for hepatitis C enables proper screening of possible carriers of the hepatitis C virus, thus enabling a reduction in virus shedding. However, being only possible if health services are prepared to deal with hepatitis C virus, through education and public awareness.

  20. Hepatitis B vaccination in preterm infants

    OpenAIRE

    Huang, F.; Lee, P; Lee, C.; Huang, L.; Chang, L; Liu, S.

    1997-01-01

    AIM—To investigate the immunogenicity and safety of existing recommendations for hepatitis B vaccination in preterm infants.
METHODS—Recombinant hepatitis B vaccine (H-B-VAX II, 5 µg per dose) was given to 85 preterm infants divided into two groups, using two different schedules. Forty four group A infants with birthweights of 

  1. The Revaccination Effect of Domestic Yeast-derived Recombinant Hepatitis B Vaccine%用国产重组酵母乙型肝炎疫苗进行再免疫的效果观察

    Institute of Scientific and Technical Information of China (English)

    王振海; 朱顺元; 吴祖云

    2001-01-01

    This paper was to observe the revaccination effect of domestic yeast-derived recombinant hepatitis B vaccine (YRHBV) to children who were born to HBsAg and HBeAg negative mothers and had been immunized with full course of blood-derived hepatitis B vaccine 4 years ago at their birth. The treated group was revaccinated with 5μg YRHBV while the control group with 10μg blood-derived HB vaccine, their anti-HBs response was determined 1,12,24 months after revaccination. The results showed that a nice anti-HBs response was discovered in group A childeren, no matter how about the anti-HBs level before revaccination was. The kinetics of anti-HBs after YDHBV revaccination was similar to that of the blood-derived HB vaccine, no statistical difference was found. So, 5μg of domestic yeast-derived recombinant HB vaccine gave same immune effect as well as 10μg blood-derived HB vaccine.%为了观察用国产重组酵母乙型肝炎(乙肝)疫苗,对已经血源乙肝疫苗初免的儿童再免疫的效果,以乙肝表面抗原(HBsAg)和乙肝e抗原(HBeAg)阴性母亲所生的、出生时全程接种血源乙肝疫苗后已4年的儿童为对象,用5μg重组酵母乙肝疫苗进行再免疫,同时用10μg血源乙肝疫苗再免疫作对照,观察2种疫苗再免疫后1、12、24个月乙肝表面抗体(抗-HBs)反应。结果显示:用5μg重组酵母乙肝疫苗进行再免疫能取得良好的抗-HBs应答,不论再免疫前抗-HBs水平高低,再免疫后能呈现免疫回忆反应。再免疫的抗体反应模式与用10μg血源乙肝疫苗进行再免疫的相似。两种乙肝疫苗再免疫的效果无显著性差异。因此,用5μg重组酵母乙肝疫苗与10μg血源乙肝疫苗再免疫能获得相同的再免疫效果。

  2. Preparation of recombinant human bone morphogenetic protein-2 loaded dextran-based microspheres and their characteristics

    Institute of Scientific and Technical Information of China (English)

    Fa-ming CHEN; Zhi-fen WU; Qin-tao WANG; Hong WU; Yong-jie ZHANG; Xin NIE; Yan JIN

    2005-01-01

    Aim: To prepare new pharmaceutical forms with sustained delivery properties of recombinant human bone morphogenetic protein-2 (rhBMP2) for tissue engineering and guided tissue regeneration (GTR) use. Methods: rhBMP2-1oaded dextranbased hydrogel microspheres (rhBMP2-MPs), which aimed to keep rhBMP2 bioactivity and to achieve long-term sustained release of rhBMP2, were prepared by double-phase emulsified condensation polymerization. The physical, chemical performances and biological characteristics of those microspheres were studied both in vitro and in vivo. Results: The microspheres' average diameter was 30.33±4.32 μm with 75.4% ranging from 20 μm to 40 μm and the drug loading and encapsulation efficiency were 7.82% and 82.25%, respectively. The rhBMP2-releasing profiles in vitro showed that rhBMP2 release could be maintained more than 10 d. The rhBMP2-MPs, with good swelling and biodegradation behavior,could be kept for 6 months at below 4 ℃ without significant characteristic change or bioactivity loss. Cytology studies showed that rhBMP2-MPs could promote the proliferation of periodontal ligament cells (PDLCs) approximately 10 d, while the bioactivity of concentrated rhBMP2 solution could keep no more than 3 d.Scanning electron microscope showed that rhBMP2-MPs could be enchased into the porous structure of calcium phosphate ceremic (CPC) and the eugonic growth of PDLCs in CPC/rhBMP2-MPs scaffolds. Animal experiments indicated that using CPC/rhBMP2-MPs scaffolds could gain more periodontal tissue regeneration than using rhBMP2 compound firsthand with CPC (CPC/rhBMP2). Conclusion:By encapsulating rhBMP2 into dextran-based microspheres, a small quantity of rhBMP2 could achieve equivalent effects to the concentrated rhBMP2 solution and at the same time, could prolong rhBMP2 retention both in vitro and in vivo.

  3. Segmentation of hepatic vessels from MRI images for planning of electroporation-based treatments in the liver

    OpenAIRE

    Marcan Marija; Pavliha Denis; Music Maja Marolt; Fuckan Igor; Magjarevic Ratko; Miklavcic Damijan

    2014-01-01

    Introduction. Electroporation-based treatments rely on increasing the permeability of the cell membrane by high voltage electric pulses delivered to tissue via electrodes. To ensure that the whole tumor is covered by the sufficiently high electric field, accurate numerical models are built based on individual patient geometry. For the purpose of reconstruction of hepatic vessels from MRI images we searched for an optimal segmentation method that would meet the following initial criteria: iden...

  4. VP22 fusion protein-based dominant negative mutant can inhibit hepatitis B virus replication

    Institute of Scientific and Technical Information of China (English)

    Jun Yi; Wei-Dong Gong; Ling Wang; Rui Ling; Jiang-Hao Chen; Jun Yun

    2005-01-01

    AIM: To investigate the inhibitory effect of VP22 fusion protein-based dominant negative (DN) mutant on Hepatitis Bvrus (HBV) replication.METHODS: Full-length or truncated fragment of VP22 was fused to C terminal of HBV core protein (HBc), and subcloned into pcDNA3.1 (-) vector, yielding eukaryotic expression plasmids of DN mutant. After transfection into HepG2.2.15 cells, the expression of DN mutant was identified by immunofluorescence staining. The inhibitory effect of DN mutant on HBV replication was indexed as the supernatant HBsAg concentration determined by RIA and HBV-DNA content by fluorescent quantification-PCR (FQ-PCR). Meanwhile, metabolism of HepG2.2.15 cells was evaluated by MTT colorimetry.RESULTS: VP22-based DN mutants and its truncated fragment were expressed in HepG2.2.15 cells, and had no toxic effect on host cells. DN mutants could inhibit HBV replication and the transduction ability of mutantbearing protein had a stronger inhibitory effect on HBV replication. DN mutants with full length of VP22 had the strongest inhibitory effect on HBV replication, reducing the HBsAg concentration by 81.94%, and the HBV-DNA content by 72.30%. MTT assay suggested that there were no significant differences in cell metabolic activity between the groups.CONCLUSION: VP22-based DN mutant can inhibit HBV replication effectively.

  5. Recombinant plasmid-based quantitative Real-Time PCR analysis of Salmonella enterica serotypes and its application to milk samples.

    Science.gov (United States)

    Gokduman, Kurtulus; Avsaroglu, M Dilek; Cakiris, Aris; Ustek, Duran; Gurakan, G Candan

    2016-03-01

    The aim of the current study was to develop, a new, rapid, sensitive and quantitative Salmonella detection method using a Real-Time PCR technique based on an inexpensive, easy to produce, convenient and standardized recombinant plasmid positive control. To achieve this, two recombinant plasmids were constructed as reference molecules by cloning the two most commonly used Salmonella-specific target gene regions, invA and ttrRSBC. The more rapid detection enabled by the developed method (21 h) compared to the traditional culture method (90 h) allows the quantitative evaluation of Salmonella (quantification limits of 10(1)CFU/ml and 10(0)CFU/ml for the invA target and the ttrRSBC target, respectively), as illustrated using milk samples. Three advantages illustrated by the current study demonstrate the potential of the newly developed method to be used in routine analyses in the medical, veterinary, food and water/environmental sectors: I--The method provides fast analyses including the simultaneous detection and determination of correct pathogen counts; II--The method is applicable to challenging samples, such as milk; III--The method's positive controls (recombinant plasmids) are reproducible in large quantities without the need to construct new calibration curves.

  6. Quantitative determination of optical and recombination losses in thin-film photovoltaic devices based on external quantum efficiency analysis

    Science.gov (United States)

    Nakane, Akihiro; Tampo, Hitoshi; Tamakoshi, Masato; Fujimoto, Shohei; Kim, Kang Min; Kim, Shinho; Shibata, Hajime; Niki, Shigeru; Fujiwara, Hiroyuki

    2016-08-01

    In developing photovoltaic devices with high efficiencies, quantitative determination of the carrier loss is crucial. In conventional solar-cell characterization techniques, however, photocurrent reduction originating from parasitic light absorption and carrier recombination within the light absorber cannot be assessed easily. Here, we develop a general analysis scheme in which the optical and recombination losses in submicron-textured solar cells are evaluated systematically from external quantum efficiency (EQE) spectra. In this method, the optical absorption in solar cells is first deduced by imposing the anti-reflection condition in the calculation of the absorptance spectrum, and the carrier extraction from the light absorber layer is then modeled by considering a carrier collection length from the absorber interface. Our analysis method is appropriate for a wide variety of photovoltaic devices, including kesterite solar cells [Cu2ZnSnSe4, Cu2ZnSnS4, and Cu2ZnSn(S,Se)4], zincblende CdTe solar cells, and hybrid perovskite (CH3NH3PbI3) solar cells, and provides excellent fitting to numerous EQE spectra reported earlier. Based on the results obtained from our EQE analyses, we discuss the effects of parasitic absorption and carrier recombination in different types of solar cells.

  7. Kinetic Evidence of Two Pathways for Charge Recombination in NiO-Based Dye-Sensitized Solar Cells.

    Science.gov (United States)

    D'Amario, Luca; Antila, Liisa J; Pettersson Rimgard, Belinda; Boschloo, Gerrit; Hammarström, Leif

    2015-03-01

    Mesoporous nickel oxide has been used as electrode material for p-type dye-sensitized solar cells (DSCs) for many years but no high efficiency cells have yet been obtained. One of the main issues that lowers the efficiency is the poor fill factor, for which a clear reason is still missing. In this paper we present the first evidence for a relation between applied potential and the charge recombination rate of the NiO electrode. In particular, we find biphasic recombination kinetics: a fast (15 ns) pathway attributed to the reaction with the holes in the valence band and a slow (1 ms) pathway assigned to the holes in the trap states. The fast component is the most relevant at positive potentials, while the slow component becomes more important at negative potentials. This means that at the working condition of the cell, the fast recombination is the most important. This could explain the low fill factor of NiO-based DSCs.

  8. Establishment of a selective evaluation method for DPP4 inhibitors based on recombinant human DPP8 and DPP9 proteins

    Directory of Open Access Journals (Sweden)

    Jinglong Liu

    2014-04-01

    Full Text Available Dipeptidyl peptidase 4 (DPP4 is recognised as an attractive anti-diabetic drug target, and several DPP4 inhibitors are already on the market. As members of the same gene family, dipeptidyl peptidase 8 (DPP8 and dipeptidyl peptidase 9 (DPP9 share high sequence and structural homology as well as functional activity with DPP4. However, the inhibition of their activities was reported to cause severe toxicities. Thus, the development of DPP4 inhibitors that do not have DPP8 and DPP9 inhibitory activity is critical for safe anti-diabetic therapy. To achieve this goal, we established a selective evaluation method for DPP4 inhibitors based on recombinant human DPP8 and DPP9 proteins expressed by Rosetta cells. In this method, we used purified recombinant 120 kDa DPP8 or DPP9 protein from the Rosetta expression system. The optimum concentrations of the recombinant DPP8 and DPP9 proteins were 30 ng/mL and 20 ng/mL, respectively, and the corresponding concentrations of their substrates were both 0.2 mmol/L. This method was highly reproducible and reliable for the evaluation of the DPP8 and DPP9 selectivity for DPP4 inhibitor candidates, which would provide valuable guidance in the development of safe DPP4 inhibitors.

  9. Multi-Homologous Recombination-Based Gene Manipulation in the Rice Pathogen Fusarium fujikuroi

    OpenAIRE

    Hwang, In Sun; Ahn, Il-Pyung

    2016-01-01

    Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this ...

  10. A bioassay based on recombinant DNA technology for determining selenium concentration.

    OpenAIRE

    Reches, M; Zhao, C; Engelberg-Kulka, H

    1994-01-01

    The trace element selenium has recently attracted attention, particularly because (i) selenocysteine is involved in the active site of various prokaryotic and eukaryotic enzymes, some of which have a role in human health; (ii) selenocysteine incorporation into these proteins is coded by UGA codons; and (iii) as a result, selenocysteine is now considered to be the 21st amino acid in an expanded genetic code. Here, we built recombinant DNA constructs in which expression of the lac'Z gene is dri...

  11. Utility value of a T-cell interferon-γ release assay based on recombinant Mycobacterium tuberculosis 11kD protein in the diagnosis of tuberculosis

    Institute of Scientific and Technical Information of China (English)

    张丽帆

    2014-01-01

    Objective To evaluate the diagnostic efficiency of a T-cell interferon-γrelease assay based on recombinant Mycobacterium tuberculosis(MTB)11kD protein for diagnosing tuberculosis.Methods This prospective study enrolled inpatients with suspected tuberculosis at PUMCH to examine the diagnostic sensitivity,specificity,predictive value(PV)and likelihood ratio(LR)of T-cell interferon-γrelease assays based on recombinant MTB-11kD

  12. Intragenotypic JFH1 based recombinant hepatitis C virus produces high levels of infectious particles but causes increased cell death

    DEFF Research Database (Denmark)

    Mateu, Guaniri; Donis, Ruben O; Wakita, Takaji;

    2008-01-01

    into the JFH1 infectious clone. All genomes produced high levels of intracellular HCV RNA and NS3 protein in Huh-7.5 transfected cells. However, JFH1 genomes containing J6 sequences from C to E2 (CE2) or C to p7 (Cp7) secreted up to 100-fold more infectious HCV particles than the parental JFH1 clone...... of the chimeric junction. Moreover, NTRNS2 a chimeric virus equivalent to the previously reported FL-J6/JFH chimera, showed a 10-fold enhancement of virus titers compared to CNS2. NTRNS2 differs from CNS2 by three nucleotide differences residing in the 5' NTR and core coding sequence and all three nucleotide...

  13. Recombination instability

    DEFF Research Database (Denmark)

    D'Angelo, N.

    1967-01-01

    A recombination instability is considered which may arise in a plasma if the temperature dependence of the volume recombination coefficient, alpha, is sufficiently strong. Two cases are analyzed: (a) a steady-state plasma produced in a neutral gas by X-rays or high energy electrons; and (b...

  14. Differential efficacy of protease inhibitors against HCV genotypes 2a, 3a, 5a, and 6a NS3/4A protease recombinant viruses

    DEFF Research Database (Denmark)

    Gottwein, Judith M; Scheel, Troels K H; Jensen, Tanja B;

    2011-01-01

    The hepatitis C virus (HCV) genotype influences efficacy of interferon (IFN)-based therapy. HCV protease inhibitors are being licensed for treatment of genotype 1 infection. Because there are limited or no data on efficacy against HCV genotypes 2-7, we aimed at developing recombinant infectious...

  15. The Analysis on Immune Effects of People Vaccinated by Hepatitis B Vaccine Made by Recombinant Deoxyribonucleic Acid Techniques in Saccharomyces Cerevisiae Yeast for 1-12 Years%接种重组乙型肝炎疫苗(酿酒酵母)后1~12年免疫效果分析

    Institute of Scientific and Technical Information of China (English)

    徐莉立; 王学文; 李永盛; 沈立萍; 王峰; 赵金华; 杨维雄; 高玉清; 唐志坚

    2013-01-01

    目的 了解1997~2008年出生儿童接种重组乙型肝炎(乙肝)疫苗(酿酒酵母)(Hepatitis B Vaccine Made by Recombinant Deoxyribonucleic Acid Techniques in Saccharomyces Cerevisiae Yeast,HepB-SCY)后的抗体水平,评价HepB-SCY的免疫持久性及保护效果.方法 在西宁市城西区、大通县和海南藏族自治州同德县,选择1997~2008年出生、有明确HepB-SCY免疫史的特定人群,每个年龄段100人左右,采集静脉血5ml,分离血清,检测乙肝病毒(Hepatitis B Virus,HBV)表面抗原、抗乙肝病毒表面抗原抗体、抗乙肝病毒核心抗原抗体三项血清学指标.结果 接种HepB-SCY后l~12年抗体阳性率保持在较高水平,抗体几何平均浓度则呈现随免疫年限延长而逐渐下降趋势.免疫人群HBV感染率为4.82%,较实施免疫前明显下降.结论 HepB-SCY免疫后效果较持久,可有效预防接种人群HBV感染.%Objective To understand the long-term immune effects of hepatitis B vaccine made by recombinant deoxyribonucleic acid techniques in saccharomyces cerevisiae yeast (HepB-SCY).Method To select the children from Chengxi,Datong and Tongde county of Qinghai province,who had been vaccinated HepB-SCY who were born from 1997 to 2008.100 children were selected each year to check their hepatitis B vaccination history and test for Hepatitis B Virus (HBV)markers.Results The positive rate of anti-hepatitis B surface antibody maintained at higher level after vaccination for 12 years,however the geometric mean concentration of anti-hepatitis B surface antibody was decreased with years.The average HBV positive rate of the children was 4.82%.It revealed significant reduction compared with the teenagers before immunization.Conclusion The long-term immune effects of HepB-SCY was satisfied and it has good effects for preventing the infection of HBV.

  16. Virus-Like Vesicle-Based Therapeutic Vaccine Vectors for Chronic Hepatitis B Virus Infection

    OpenAIRE

    Tracy D Reynolds; Buonocore, Linda; Rose, Nina F.; Rose, John K.; Robek, Michael D.

    2015-01-01

    More than 500,000 people die each year from the liver diseases that result from chronic hepatitis B virus (HBV) infection. Therapeutic vaccines, which aim to elicit an immune response capable of controlling the virus, offer a potential new treatment strategy for chronic hepatitis B. Recently, an evolved, high-titer vaccine platform consisting of Semliki Forest virus RNA replicons that express the vesicular stomatitis virus glycoprotein (VSV G) has been described. This platform generates virus...

  17. Visualization of Gene Mutation Complicated Pattern of Hepatitis B Virus Based on Cellular Automata

    Institute of Scientific and Technical Information of China (English)

    SHAO Shi-huang; XIAO Xuan; DING Yong-sheng; HUANG Zhen-de

    2005-01-01

    Hepatitis B virus shows instantaneous and high rate mutations in biological experiments, some sorts of which affect the efficiency of virus replication greatly through enhancing or depressing the viral replication, while others have no influence at all. Taking advantage of prominent features of cellular automata, we simulate the effect of hepatitis B virus gene mutation on its replication efficiency. The computer simulation results demonstrate the feasibility of our novel model by comparing with the results of biological experiments.

  18. A new vector for recombination-based cloning of large DNA fragments from yeast artificial chromosomes.

    OpenAIRE

    Bradshaw, M S; Bollekens, J A; Ruddle, F H

    1995-01-01

    The functional analysis of genes frequently requires manipulation of large genomic regions embedded in yeast artificial chromosomes (YACs). We have designed a yeast-bacteria shuttle vector, pClasper, that can be used to clone specific regions of interest from YACs by homologous recombination. The important feature of pClasper is the presence of the mini-F factor replicon. This leads to a significant increase in the size of the plasmid inserts that can be maintained in bacteria after cloning b...

  19. Plant Cell-Based Recombinant Antibody Manufacturing with a 200 L Orbitally Shaken Disposable Bioreactor.

    Science.gov (United States)

    Raven, Nicole; Schillberg, Stefan; Rasche, Stefan

    2016-01-01

    Tobacco BY-2 cells are an attractive platform for the manufacture of a variety of biopharmaceutical proteins, including antibodies. Here, we describe the scaled-up cultivation of human IgG-secreting BY-2 cells in a 200 L orbitally shaken disposable bioreactor, resulting in cell growth and recombinant protein yields that are proportionately comparable with those obtained from cultivations in 500 mL shake flasks. Furthermore, we present an efficient downstream process for antibody recovery from the viscous spent culture medium using expanded bed adsorption (EBA) chromatography. PMID:26614289

  20. Optimisation of prime-boost immunization in mice using novel protein-based and recombinant vaccinia (Tiantan-based HBV vaccine.

    Directory of Open Access Journals (Sweden)

    Hong Chen

    Full Text Available BACKGROUND: A therapeutic vaccine for chronic hepatitis B virus (HBV infection that enhances virus-specific cellular immune responses is urgently needed. The "prime-boost" regimen is a widely used vaccine strategy against many persistence infections. However, few reports have addressed this strategy applying for HBV therapeutic vaccine development. METHODOLOGY/PRINCIPAL FINDINGS: To develop an effective HBV therapeutic vaccine, we constructed a recombinant vaccinia virus (Tiantan containing the S+PreS1 fusion antigen (RVJSS1 combined with the HBV particle-like subunit vaccine HBVSS1 to explore the most effective prime-boost regimen against HBV. The immune responses to different prime-boost regimens were assessed in C57BL/C mice by ELISA, ELISpot assay and Intracellular cytokine staining analysis. Among the combinations tested, an HBV protein particle vaccine priming and recombinant vaccinia virus boosting strategy accelerated specific seroconversion and produced high antibody (anti-PreS1, anti-S antibody titres as well as the strongest multi-antigen (PreS1, and S-specific cellular immune response. HBSS1 protein prime/RVJSS1 boost immunization was also generated more significant level of both CD4+ and CD8+ T cell responses for Th1 cytokines (TNF-α and IFN-γ. CONCLUSIONS: The HBSS1 protein-vaccine prime plus RVJSS1 vector boost elicits specific antibody as well as CD4 and CD8 cells secreting Th1-like cytokines, and these immune responses may be important parameters for the future HBV therapeutic vaccines.

  1. Construction and expression of a recombinant eukaryotic expression plasmid containing the preS1-preS2-S genes of hepatitis B virus and the granulocyte-macrophage colony stimulating factor gene: A study of its immunomodulatory effects

    OpenAIRE

    GONG, JUN-YUAN; Liu, Xin; Dong, Yan; ZHOU, TIAN-HONG; LI, JUN-WU

    2012-01-01

    A total of 10–20% of the population remains unresponsive or weakly responsive to hepatitis B vaccine, which is composed of hepatitis B surface antigen HBsAg (S protein). Therefore, it is necessary to develop a hepatitis B vaccine with a better penetrating and responsive rate. In the present study, a plasmid pVAX1-L-GM was constructed and its immunomodulatory effect of as hepatitis B virus (HBV) DNA vaccine was analyzed through the immunization of BALB/c mice. Immune responses were measured af...

  2. L-Lactate-selective microbial sensor based on flavocytochrome b2-enriched yeast cells using recombinant and nanotechnology approaches.

    Science.gov (United States)

    Karkovska, Maria; Smutok, Oleh; Stasyuk, Nataliya; Gonchar, Mykhailo

    2015-11-01

    In the recent years, nanotechnology is the most developing branch due to a wide variety of potential applications in biomedical, biotechnological and agriculture fields. The binding nanoparticles with various biological molecules makes them attractive candidates for using in sensor technologies. The particularly actual is obtaining the bionanomembranes based on biocatalytic elements with improved sensing characteristics. The aim of this investigation is to study the properties of microbial L-lactate-selective sensor based on using the recombinant Hansenula polymorpha yeast cells overproducing flavocytochrome b2 (FC b2), as well as additionally enriched by the enzyme bound with gold nanoparticles (FC b2-nAu). Although, the high permeability of the living cells to nanoparticles is being intensively studied (mostly for delivery of drugs), the idea of using both recombinant technology and nanotechnology to increase the amount of the target enzyme in the biosensing layer is really novel. The FC b2-nAu-enriched living and permeabilized yeast cells were used for construction of a bioselective membrane of microbial L-lactate-selective amperometric biosensor. Phenazine methosulphate was served as a free defusing electron transfer mediator which provides effective electron transfer from the reduced enzyme to the electrode surface. It was shown that the output to L-lactate of FC b2-nAu-enriched permeabilized yeast cells is 2.5-fold higher when compared to the control cells. The obtained results confirm that additional enrichment of the recombinant yeast cell by the enzyme bound with nanoparticles improves the analytical parameters of microbial sensor.

  3. Support vector machine for classification of meiotic recombination hotspots and coldspots in Saccharomyces cerevisiae based on codon composition

    Directory of Open Access Journals (Sweden)

    Sun Xiao

    2006-04-01

    Full Text Available Abstract Background Meiotic double-strand breaks occur at relatively high frequencies in some genomic regions (hotspots and relatively low frequencies in others (coldspots. Hotspots and coldspots are receiving increasing attention in research into the mechanism of meiotic recombination. However, predicting hotspots and coldspots from DNA sequence information is still a challenging task. Results We present a novel method for classification of hot and cold ORFs located in hotspots and coldspots respectively in Saccharomyces cerevisiae, using support vector machine (SVM, which relies on codon composition differences. This method has achieved a high classification accuracy of 85.0%. Since codon composition is a fusion of codon usage bias and amino acid composition signals, the ability of these two kinds of sequence attributes to discriminate hot ORFs from cold ORFs was also investigated separately. Our results indicate that neither codon usage bias nor amino acid composition taken separately performed as well as codon composition. Moreover, our SVM based method was applied to the full genome: We predicted the hot/cold ORFs from the yeast genome by using cutoffs of recombination rate. We found that the performance of our method for predicting cold ORFs is not as good as that for predicting hot ORFs. Besides, we also observed a considerable correlation between meiotic recombination rate and amino acid composition of certain residues, which probably reflects the structural and functional dissimilarity between the hot and cold groups. Conclusion We have introduced a SVM-based novel method to discriminate hot ORFs from cold ones. Applying codon composition as sequence attributes, we have achieved a high classification accuracy, which suggests that codon composition has strong potential to be used as sequence attributes in the prediction of hot and cold ORFs.

  4. Fusion protein-based biofilm fabrication composed of recombinant azurin–myoglobin for dual-level biomemory application

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Taek [Research Institute for Basic Science, Sogang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Chung, Yong-Ho; Yoon, Jinho [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of); Min, Junhong [School of Integrative Engineering, Chung-Ang University, Heukseok-dong, Dongjak-gu, Seoul 156-756 (Korea, Republic of); Choi, Jeong-Woo, E-mail: jwchoi@sogang.ac.kr [Department of Chemical and Biomolecular Engineering, Sogang University, Heukseok-dong, Dongjak-gu, 35 Baekbeom-ro (Sinsu-dong), Mapo-gu, Seoul 121-742 (Korea, Republic of)

    2014-11-30

    Graphical abstract: - Highlights: • We developed the fusion protein-based biofilm on the inorganic surface. • For making the fusion protein, the recombinant azurin and the myoglobin was conjugated by the native chemical ligation method. • The developed fusion protein shows unique electrochemical property. • The proposed fusion protein biofilm appears to be a good method for dual-level biomemory device. - Abstract: In the present study, a fusion protein-based biofilm composed of a recombinant azurin–myoglobin (Azu-Myo) has been developed and confirmed its original electrochemical property for dual-level biomemory device application. For this purpose, the azurin was modified with cysteine residues for direct immobilization and conjugation. Then, the recombinant azurin was conjugated with the myoglobin via a sulfo-SMCC bifunctional linker using the chemical ligation method (CLM). The SDS-PAGE and UV–vis spectroscopy were performed to examine the fusion protein conjugates. The prepared Azu-Myo fusion protein was self-assembled onto Au substrate for the biofilm fabrication. Then, the atomic force microscopy (AFM) was used to confirm the immobilization and the surface-enhanced Raman spectroscopy (SERS) was carried out to the surface analysis. Also, the cyclic voltammetry (CV) was carried out to observe an electrochemical property of fabricated biofilm. As a result, the two pair of redox potential values was obtained for dual-level biomemory device application. Then, the dual-level biomemory function was verified by the multi-potential chronoamperometry (MPCA). The results indicate a new fabrication method and material combination for advances in bioelectronic device development.

  5. Detection of Q Fever Specific Antibodies Using Recombinant Antigen in ELISA with Peroxidase Based Signal Amplification

    Directory of Open Access Journals (Sweden)

    Hua-Wei Chen

    2014-01-01

    Full Text Available Currently, the accepted method for Q fever serodiagnosis is indirect immunofluorescent antibody assay (IFA using the whole cell antigen. In this study, we prepared the recombinant antigen of the 27-kDa outer membrane protein (Com1 which has been shown to be recognized by Q fever patient sera. The performance of recombinant Com1 was evaluated in ELISA by IFA confirmed serum samples. Due to the low titers of IgG and IgM in Q fever patients, the standard ELISA signals were further amplified by using biotinylated anti-human IgG or IgM plus streptavidin-HRP polymer. The modified ELISA can detect 88% (29 out of 33 of Q fever patient sera collected from Marines deployed to Iraq. Less than 5% (5 out of 156 of the sera from patients with other febrile diseases reacted with the Com1. These results suggest that the modified ELISA using Com1 may have the potential to improve the detection of Q fever specific antibodies.

  6. A model-based meta-analysis of sofosbuvir-based treatments in chronic hepatitis C patients.

    Science.gov (United States)

    Pérez-Pitarch, Alejandro; Guglieri-López, Beatriz; Ferriols-Lisart, Rafael; Merino-Sanjuán, Matilde

    2016-03-01

    The objective of this study was to compare the efficacy of sofosbuvir-based treatments in patients with chronic hepatitis C virus (HCV) infection using a model-based meta-analysis (MBMA). A bibliographic search was performed to identify clinical trials involving sofosbuvir as a unique direct-acting antiviral (DAA) agent or together with daclatasvir, ledipasvir or simeprevir for the treatment of diagnosed HCV infection. The time course of the virological response (VR) was modelled to estimate the effect of treatment and the influence of population characteristics on the longitudinal efficacy profile. The model was validated and simulations of 10 different treatment schedules were performed. Data from 19 clinical trials were included in the analysis. According to the developed model, therapy with sofosbuvir+ledipasvir is the most effective therapy in all scenarios, but it does not differ greatly in terms of sustained VR with respect to other combinations of DAA treatments. In conclusion, this MBMA generates knowledge regarding hypothetical head-to-head trials that have not been conducted previously. Therapies with sofosbuvir+ledipasvir are probably the most effective sofosbuvir-based treatments.

  7. Should patients with abnormal liver function tests in primary care be tested for chronic viral hepatitis: cost minimisation analysis based on a comprehensively tested cohort

    Directory of Open Access Journals (Sweden)

    Lilford Richard J

    2011-03-01

    Full Text Available Abstract Background Liver function tests (LFTs are ordered in large numbers in primary care, and the Birmingham and Lambeth Liver Evaluation Testing Strategies (BALLETS study was set up to assess their usefulness in patients with no pre-existing or self-evident liver disease. All patients were tested for chronic viral hepatitis thereby providing an opportunity to compare various strategies for detection of this serious treatable disease. Methods This study uses data from the BALLETS cohort to compare various testing strategies for viral hepatitis in patients who had received an abnormal LFT result. The aim was to inform a strategy for identification of patients with chronic viral hepatitis. We used a cost-minimisation analysis to define a base case and then calculated the incremental cost per case detected to inform a strategy that could guide testing for chronic viral hepatitis. Results Of the 1,236 study patients with an abnormal LFT, 13 had chronic viral hepatitis (nine hepatitis B and four hepatitis C. The strategy advocated by the current guidelines (repeating the LFT with a view to testing for specific disease if it remained abnormal was less efficient (more expensive per case detected than a simple policy of testing all patients for viral hepatitis without repeating LFTs. A more selective strategy of viral testing all patients for viral hepatitis if they were born in countries where viral hepatitis was prevalent provided high efficiency with little loss of sensitivity. A notably high alanine aminotransferase (ALT level (greater than twice the upper limit of normal on the initial ALT test had high predictive value, but was insensitive, missing half the cases of viral infection. Conclusions Based on this analysis and on widely accepted clinical principles, a "fast and frugal" heuristic was produced to guide general practitioners with respect to diagnosing cases of viral hepatitis in asymptomatic patients with abnormal LFTs. It recommends

  8. Hepatitis B Vaccine

    Science.gov (United States)

    ... as a combination product containing Hepatitis A Vaccine, Hepatitis B Vaccine) ... Hepatitis B vaccine: Why get vaccinated?Hepatitis B vaccine can prevent hepatitis B, and the serious consequences of hepatitis ...

  9. Hepatitis A

    Science.gov (United States)

    ... an inflammation of the liver. One type, hepatitis A, is caused by the hepatitis A virus (HAV). The disease spreads through contact with ... washed in untreated water Putting into your mouth a finger or object that came into contact with ...

  10. Viral Hepatitis

    Science.gov (United States)

    ... Hepatitis viruses B and C can cause both acute and chronic infections. Chronic hepatitis B and C are serious health problems. They can lead to: Cirrhosis (suh-ROH-suhs) Liver failure Liver cancer Return to top How is viral ...

  11. Characterization of Biosensors Based on Recombinant Glutamate Oxidase: Comparison of Crosslinking Agents in Terms of Enzyme Loading and Efficiency Parameters.

    Science.gov (United States)

    Ford, Rochelle; Quinn, Susan J; O'Neill, Robert D

    2016-01-01

    Amperometric l-glutamate (Glu) biosensors, based on both wild-type and a recombinant form of l-glutamate oxidase (GluOx), were designed and characterized in terms of enzyme-kinetic, sensitivity and stability parameters in attempts to fabricate a real-time Glu monitoring device suitable for future long-term detection of this amino acid in biological and other complex media. A comparison of the enzyme from these two sources showed that they were similar in terms of biosensor performance. Optimization of the loading of the polycationic stabilization agent, polyethyleneimine (PEI), was established before investigating a range of crosslinking agents under different conditions: glutaraldehyde (GA), polyethylene glycol (PEG), and polyethylene glycol diglycidyl ether (PEGDE). Whereas PEI-free biosensor designs lost most of their meager Glu sensitivity after one or two days, configurations with a 2:5 ratio of dip-evaporation applications of PEI(1%):GluOx(400 U/mL) displayed a 20-fold increase in their initial sensitivity, and a decay half-life extended to 10 days. All the crosslinkers studied had no effect on initial Glu sensitivity, but enhanced biosensor stability, provided the crosslinking procedure was carried out under well-defined conditions. The resulting biosensor design based on the recombinant enzyme deposited on a permselective layer of poly-(ortho-phenylenediamine), PoPD/PEI₂/GluOx₅/PEGDE, displayed good sensitivity (LOD term monitoring of Glu concentration dynamics in complex media.

  12. Anti-HBs levels in infants of hepatitis B carrier mothers after delayed active immunization with recombinant vaccine concomitant with DTP-polio vaccine: Is there need for a second dose of HBIg?

    NARCIS (Netherlands)

    P.M. Grosheide (Pia Maria); R. Del Canho (R.); M. Voogd (M.); R.A. Heijtink; S.W. Schalm (Solko)

    1994-01-01

    textabstractThe need for an additional dose of hepatitis B immune globulin (HBIg) was studied by comparing infants receiving 1 ml HBIg at birth followed by hepatitis B immunization, concomitant with DTP-polio vaccine, at 3, 4, 5 and 11 months (schedule E), with infants receiving the same schedule wi

  13. Laminin-511 and laminin-521-based matrices for efficient hepatic specification of human pluripotent stem cells.

    Science.gov (United States)

    Kanninen, Liisa K; Harjumäki, Riina; Peltoniemi, Pasi; Bogacheva, Mariia S; Salmi, Tuuli; Porola, Pauliina; Niklander, Johanna; Smutný, Tomáš; Urtti, Arto; Yliperttula, Marjo L; Lou, Yan-Ru

    2016-10-01

    Human pluripotent stem cells (hPSCs) have gained a solid foothold in basic research and drug industry as they can be used in vitro to study human development and have potential to offer limitless supply of various somatic cell types needed in drug development. Although the hepatic differentiation of hPSCs has been extensively studied, only a little attention has been paid to the role of the extracellular matrix. In this study we used laminin-511, laminin-521, and fibronectin, found in human liver progenitor cells, as culture matrices for hPSC-derived definitive endoderm cells. We observed that laminin-511 and laminin-521 either alone or in combination support the hepatic specification and that fibronectin is not a vital matrix protein for the hPSC-derived definitive endoderm cells. The expression of the laminin-511/521-specific integrins increased during the definitive endoderm induction and hepatic specification. The hepatic cells differentiated on laminin matrices showed the upregulation of liver-specific markers both at mRNA and protein levels, secreted human albumin, stored glycogen, and exhibited cytochrome P450 enzyme activity and inducibility. Altogether, we found that laminin-511 and laminin-521 can be used as stage-specific matrices to guide the hepatic specification of hPSC-derived definitive endoderm cells. PMID:27372423

  14. Molecular Characterization and SYBR Green Ⅰ-Based Quantitative PCR for Duck Hepatitis Virus Type 1

    Institute of Scientific and Technical Information of China (English)

    LUO Yu-jun; ZHANG Gui-hong; XU Xiao-qin; CHEN Jian-hong; LIAO Ming

    2008-01-01

    To determine the genomic sequence of a duck hepatitis virus type 1 (DHV-1) strain,real-time quantitative polyrnerase chain reaction (RTQ-PCR) assay based on SYBR Green Ⅰ technology was developed to target 3D gene of DHV-1.Comparative sequence analysis showed that the genome has a typical picornarivus genetic organization,and strain DHV-1 R genetic organaiztion is 5' untranslated region (UTR)-VPO-VP3-VP1-2A1-2A2-2B-2C-3A-3B-3C-3D-3' UTR,DHV-1 R has close relationship with Parechovirus,and has 95.1-99.1% nucleotide sequence identity with other DHV-1 strains.Based on the DHV-1 sequences in GenBank,three pairs of specific primers were designed to amplify DHV-1 using real-time PCR.The results showed that real-time PCR Tm value is 85.6℃ and the real-time PCR provides a broad dynamic range,detecting from 102 to 109 copies of DHV-1 cDNA per reaction.No cross-reactions were found in specimens containing DPV,AIV and NDV.It is concluded that DHV-1 belongs to a new group of the family Picornaviridae that may form a separate genus most closely related to the genus Parechovirus.All results showed that the real-time PCR has high sensitivity and specificity to detect DHV-1 using SYBR Green Ⅰ dissociation curve analysis,isolates can be distinguished by their melting temperature.These methods are rapid,sensitive,and reliable,and can be readily adapted for detection of DHV-1 from other clinical samples.

  15. Recombinant vesicular stomatitis virus-based dengue-2 vaccine candidate induces humoral response and protects mice against lethal infection.

    Science.gov (United States)

    Lauretti, Flavio; Chattopadhyay, Anasuya; de Oliveira França, Rafael Freitas; Castro-Jorge, Luiza; Rose, John; Fonseca, Benedito A L da

    2016-09-01

    Dengue is the most important arbovirus disease throughout the world and it is responsible for more than 500,000 dengue hemorrhagic cases and 22,000 deaths every year. One vaccine was recently licensed for human use in Brazil, Mexico and Philippines and although at least seven candidates have been in clinical trials the results of the most developed CYD vaccine have demonstrated immunization problems, such as uneven protection and interference between serotypes. We constructed a vaccine candidate based on vesicular stomatitis virus (VSV) expression of pre-membrane (prM) and envelope (E) proteins of dengue-2 virus (DENV-2) and tested it in mice to evaluate immunogenicity and protection against DENV-2 infection. VSV has been successfully used as vaccine vectors for several viruses to induce strong humoral and cellular immune responses. The VSV-DENV-2 recombinant was constructed by inserting the DENV-2 structural proteins into a VSV plasmid DNA for recombinant VSV-DENV-2 recovery. Infectious recombinant VSV viruses were plaque purified and prM and E expression were confirmed by immunofluorescence and radiolabeling of proteins of infected cells. Forty Balb/C mice were inoculated through subcutaneous (s.c.) route with VSV-DENV-2 vaccine in a two doses schedule 15 d apart and 29 d after first inoculation, sera were collected and the mice were challenged with 50 lethal doses (LD50) of a neurovirulent DENV-2. The VSV-DENV-2 induced anti-DENV-2 antibodies and protected animals in the challenge experiment comparable to DENV-2 immunization control group. We conclude that VSV is a promising platform to test as a DENV vaccine and perhaps against others Flaviviridae.

  16. Bacterially produced recombinant influenza vaccines based on virus-like particles.

    Directory of Open Access Journals (Sweden)

    Andrea Jegerlehner

    Full Text Available Although current influenza vaccines are effective in general, there is an urgent need for the development of new technologies to improve vaccine production timelines, capacities and immunogenicity. Herein, we describe the development of an influenza vaccine technology which enables recombinant production of highly efficient influenza vaccines in bacterial expression systems. The globular head domain of influenza hemagglutinin, comprising most of the protein's neutralizing epitopes, was expressed in E. coli and covalently conjugated to bacteriophage-derived virus-like particles produced independently in E.coli. Conjugate influenza vaccines produced this way were used to immunize mice and found to elicit immune sera with high antibody titers specific for the native influenza hemagglutinin protein and high hemagglutination-inhibition titers. Moreover vaccination with these vaccines induced full protection against lethal challenges with homologous and highly drifted influenza strains.

  17. Hepatitis C

    Science.gov (United States)

    ... an inflammation of the liver. One type, hepatitis C, is caused by the hepatitis C virus (HCV). It usually spreads through contact with ... childbirth. Most people who are infected with hepatitis C don't have any symptoms for years. If ...

  18. Non-genotype-specific role of the hepatitis C virus 5' untranslated region in virus production and in inhibition by interferon

    DEFF Research Database (Denmark)

    Li, Yi-Ping; Ramirez, Santseharay; Gottwein, Judith M;

    2011-01-01

    The 5' untranslated region (5'UTR) of hepatitis C virus (HCV) is structured into four domains (I-IV) with numerous genotype-specific nucleotides. It is unknown whether the polymorphisms confer genotype-specific functions to the 5'UTR. Using viable JFH1-based Core-NS2 recombinants, we developed an...

  19. 不同剂次重组乙型病毒性肝炎疫苗(汉逊酵母)加强免疫效果研究%Evaluation on booster immunization efficacy of 10 μg recombinant hepatitis B vaccine made by recombinant Deoxyribonucleic Acid Techniques in Hansenula Polymorpha Yeast by different dosage in children

    Institute of Scientific and Technical Information of China (English)

    沈灵智; 陈永弟; 蒋征刚; 李倩; 陈恩富; 梁晓峰; 崔富强; 姚军

    2011-01-01

    Objective To analyze the efficacy of booster immunization with 10 μg hepatitis B vaccine made by recombinant deoxyribonucleic acid ( DNA) techniques in Hansenula Polymorpha Yeast ( HepB-HPY) by different dosage. Methods Totally 1981 children aged over 5 years were selected, who completed the basic immunization of hepatitis B vaccine at birth, the blood plasma specimens of all sampled children were detected for hepatitis B virus (HBV) surface antigen (HBsAg), antibody to hepatitis B virus surface antigen ( Anti-HBs) and antibody to hepatitis B virus core antigen (Anti-HBc) by chemiluminescence assay. Then, they were classified into Anti-HBs positive group and negative groups The children in positive group were immunized with one dose of 10 μg HepB-HPY, while those in negative group were immunized with three doses of 10 μg HepB-HPY. Their blood samples were collected after 1 month to detected Anti-HBs. Results The Anti-HBs positive rates were 38. 62% , 95. 66% and 99. 75% respectively before booster immunization, after booster immunization with one dosage and after booster immunization with three dosages, the difference was statistical significant(x2 = 73. 794 - 1736. 11, all P<0. 05). For the negative group, the Anti-HBs positive conversion rate after booster immunization with one dosage and three dosages were 92. 93% and 99. 67% respectively, the significant difference was observed (x2 = 77. 582, P<0. 05), while the Geometric Mean Concentration (GMC) of Anti-HBs negative children immunized with one dosage and three dosages were 783. 23 mIU/ml and 2463. 97 mIU/ml respectively, there was statistical significant difference (t = - 14. 201, P<0. 05). Compared with the children with Anti-HBs titers of< 0 - 1 mIU/ml before booster immunization, the positive conversion rate and GMC were significantly higher in those with the titers of 1 - 10 mIU/ml after booster immunization (all P < 0. 05). Conclusion It is suitable to use 10 μg HepB-HPY in Anti-HBs negative

  20. Hepatitis C virus infection and risk of cancer: a population-based cohort study

    DEFF Research Database (Denmark)

    Omland, Lars; Farkas, Dora Körmendiné; Jepsen, Peter;

    2010-01-01

    Hepatitis C virus (HCV) infection is associated with an increased risk of primary liver cancer; however, 5- and 10-year risk estimates are needed. The association of HCV with non-Hodgkin lymphoma (NHL) is uncertain and the association with other cancers is unknown.......Hepatitis C virus (HCV) infection is associated with an increased risk of primary liver cancer; however, 5- and 10-year risk estimates are needed. The association of HCV with non-Hodgkin lymphoma (NHL) is uncertain and the association with other cancers is unknown....

  1. Model-based projections of the population-level impact of hepatitis A vaccination in Mexico

    Science.gov (United States)

    Van Effelterre, Thierry; De Antonio-Suarez, Rodrigo; Cassidy, Adrian; Romano-Mazzotti, Luis; Marano, Cinzia

    2012-01-01

    There are indications of a shift in the pattern of hepatitis A (HAV) in Mexico from high to intermediate endemicity, progressively increasing the mean age of infection and the proportion of cases which are symptomatic. This study estimated the potential impact of universal infant HAV vaccination in Mexico with two doses of Havrix™ at 12 and 18 mo of age on all HAV infections and symptomatic HAV infections. We developed a dynamic transmission model that accounts for changes in demography and HAV epidemiology. It was calibrated using Mexican age-specific seroprevalence and symptomatic HAV incidence data. With 70% first-dose coverage and 85% second-dose coverage, the calibrated model projected that HAV vaccination would reduce the incidence of all HAV infections (symptomatic and asymptomatic) after the first 25 y of vaccination by 71–76% (minimum and maximum for different transmission scenarios). The projected reduction in cumulative incidence of symptomatic HAV infections over the first 25 y of vaccination was 45–51%. With 90% first-dose coverage and 85% second-dose coverage, the projected reduction in incidence of all HAV infections was 85–93%, and the projected reduction in the cumulative incidence of symptomatic HAV infections was 61–67%, over a 25-y time frame. Sensitivity analyses indicated that second-dose coverage is important under the conservative base-case assumptions made about the duration of vaccine protection. The model indicated that universal infant HAV vaccination could substantially reduce the burden of HAV disease in Mexico. PMID:22854667

  2. Recombination structure and genetic relatedness among members of the family Bromoviridae based on their RNAs 1 and 2 sequence analyses.

    Science.gov (United States)

    Boulila, Moncef

    2009-06-01

    In determining putative recombination events and their evolution rates in the RNAs 1 and 2 of currently the known members of the family Bromoviridae, a detailed study comprising 107 accessions retrieved from the international databases, has been carried out by using RECCO and RDP v3.31beta algorithms. These programs allowed the detection of potential recombination sites in all the five virus genera composing the family Bromoviridae with various degrees of consistency. The RNAs 1 and 2 showed inferred phylogenies fully congruent and clearly delineated five clusters representing the five studied virus genera. In this respect, we proposed to classify the Ilarviruses in three distinct subgroups instead of 10 as mentioned in several reports of the International Committee on Taxonomy of Viruses where its suggestions were based on antigenic differences. Moreover, we confirmed that Alfalfa mosaic virus should be considered as a component of the Ilarvirus genus instead of being the unique representative of Alfamovirus genus. In addition, Pelargonium zonate spot and Olive latent 2 viruses fully deserve their affiliation to the family Bromoviridae. PMID:19255837

  3. A novel color image encryption algorithm based on genetic recombination and the four-dimensional memristive hyperchaotic system

    Science.gov (United States)

    Chai, Xiu-Li; Gan, Zhi-Hua; Lu, Yang; Zhang, Miao-Hui; Chen, Yi-Ran

    2016-10-01

    Recently, many image encryption algorithms based on chaos have been proposed. Most of the previous algorithms encrypt components R, G, and B of color images independently and neglect the high correlation between them. In the paper, a novel color image encryption algorithm is introduced. The 24 bit planes of components R, G, and B of the color plain image are obtained and recombined into 4 compound bit planes, and this can make the three components affect each other. A four-dimensional (4D) memristive hyperchaotic system generates the pseudorandom key streams and its initial values come from the SHA 256 hash value of the color plain image. The compound bit planes and key streams are confused according to the principles of genetic recombination, then confusion and diffusion as a union are applied to the bit planes, and the color cipher image is obtained. Experimental results and security analyses demonstrate that the proposed algorithm is secure and effective so that it may be adopted for secure communication. Project supported by the National Natural Science Foundation of China (Grant Nos. 61203094 and 61305042), the Natural Science Foundation of the United States (Grant Nos. CNS-1253424 and ECCS-1202225), the Science and Technology Foundation of Henan Province, China (Grant No. 152102210048), the Foundation and Frontier Project of Henan Province, China (Grant No. 162300410196), the Natural Science Foundation of Educational Committee of Henan Province, China (Grant No. 14A413015), and the Research Foundation of Henan University, China (Grant No. xxjc20140006).

  4. CLINICAL EVALUATION OF FOUR RECOMBINANT TREPONEMA PALLIDUM ANTIGEN-BASED RAPID TESTS IN THE DIAGNOSIS OF SYPHILIS

    Institute of Scientific and Technical Information of China (English)

    Lin-na Wang; Lei Yang; He-yi Zheng

    2007-01-01

    To assess the sensitivity, specificity, and feasibility of 4 recombinant Treponema pallidum antigenbased rapid tests in the diagnosis of syphilis.Methods A total of 970 outpatients were selected from the Sexually Transmitted Diseases Centre of Peking Union Medical College Hospital. Venous blood was collected and serum was extracted. T. pallidum antibodies in whole blood, anticoagulant whole blood, and serum were detected using 4 recombinant T. pallidum antigen-based rapid tests.T. pallidum haemagglutination test (TPHA) was considered as the gold standard for the detection of T. pallidum specific antibodies in serum. The sensitivities and specificities of four methods were analyzed.Results The sensitivities and specificities of Abbott Determine Syphilis TP test, SD-BIOLINE Syphilis 3.0 test,VISITECT-SYPHILIS test, and Syphicheck-WB test for serum specimens were 100% and 98.9%, 95.7% and 98.0%, 94.6% and 98.2%, 68.1% and 98.9%; for whole blood were 74.1% and 99.5%, 87.9% and 99.4%,73.2% and 99.7%, 64.7% and 99.7%. The observed sensitivities of the 4 rapid diagnosis tests were not significantly different with TPHA ( P>0.05 ).Conclusions The 4 rapid tests show good performance and characteristics in the diagnosis of syphilis. Furthermore, they are more sensitive for serum specimens than whole blood.

  5. Hypoksisk hepatitis

    DEFF Research Database (Denmark)

    Amadid, Hanan; Schiødt, Frank Vinholt

    2014-01-01

    Hypoxic hepatitis (HH), also known as ischaemic hepatitis or shock liver, is an acute liver injury caused by hepatic hypoxia. Cardiac failure, respiratory failure and septic shock are the main underlying conditions. In each of these conditions, several haemodynamic mechanisms lead to hepatic...... hypoxia. A shock state is observed in only 50% of cases. Thus, shock liver and ischaemic hepatitis are misnomers. HH can be a diagnostic pitfall but the diagnosis can be established when three criteria are met. Prognosis is poor and prompt identification and treatment of the underlying conditions...

  6. Association of Sjögrens Syndrome in Patients with Chronic Hepatitis Virus Infection: A Population-Based Analysis

    Science.gov (United States)

    Yeh, Chih-Ching; Wang, Wen-Chang; Wu, Chien-Sheng; Sung, Fung-Chang; Su, Chien-Tien; Shieh, Ying-Hua; Chang, Shih-Ni; Su, Fu-Hsiung

    2016-01-01

    Objective The association between Sjögren’s syndrome (SS) and chronic hepatitis virus infection is inconclusive. Hepatitis B (HBV) and hepatitis C virus (HCV) infections are highly prevalent in Taiwan. We used a population-based case-control study to evaluate the associations between SS and HBV and HCV infections. Materials and Methods We identified 9,629 SS patients without other concomitant autoimmune diseases and 38,516 sex- and age-matched controls without SS from the Taiwan National Health Insurance claims data between 2000 and 2011. We utilized multivariate logistic regression to estimate the odds ratios (ORs) and 95% confidence intervals (CIs) of the associations between SS and HBV and HCV infections. Sex- and age-specific (<55 and ≥55 years) risks of SS were evaluated. Results The risk of SS was higher in patients with HCV than in those without chronic viral hepatitis (OR = 2.49, 95% CI = 2.16–2.86). Conversely, HBV infection was not associated with SS (OR = 1.10, 95% CI = 0.98–1.24). Younger HCV patients were at a higher risk for SS (<55 years: OR = 3.37, 95% CI = 2.62–4.35; ≥55 years: OR = 2.20, 95% CI = 1.84–2.62). Men with HCV were at a greater risk for SS (women: OR = 2.26, 95% CI = 1.94–2.63; men: OR = 4.22, 95% CI = 2.90–6.16). Only men with chronic HBV exhibited a higher risk of SS (OR = 1.61, 95% CI = 1.21–2.14). Conclusion HCV infection was associated with SS; however, HBV only associated with SS in men. PMID:27560377

  7. 20-years of population-based cancer registration in hepatitis B and liver cancer prevention in the Gambia, West Africa.

    Directory of Open Access Journals (Sweden)

    Ebrima Bah

    Full Text Available BACKGROUND: The Gambia Hepatitis Intervention Study (GHIS was designed as a randomised control trial of infant hepatitis B vaccination applied to public health policy, with the main goal of preventing primary liver cancer later in adult life in The Gambia. To that effect, the National Cancer Registry of The Gambia (NCR, a population-based cancer registry (PBCR, was established in 1986 to actively collect data on all cancer diagnosis nation-wide. We extracted 20-years (1990-2009 of data to assess for the first time, the evolution of the most common cancers, also describe and demonstrate the role of the PBCR in a hepatitis B and liver cancer prevention programme in this population. METHODS AND FINDINGS: We estimated Age-Standardised Incidence Rates (ASR (W of the most common cancers registered during the period by gender. The registration period was divided into four 5-year intervals and incidence rates were estimated for each interval. The most common cancers in males were liver, prostate, lung plus bronchus, non-Hodgkin lymphoma (NHL and stomach, accounting for 60%, 5%, 4%, 5% and 3%, respectively. Similarly, cancers of the cervix uteri, liver, breast and NHL, were the most common in females, accounting for 33%, 24%, 11% and 4% of the female cancers, respectively. CONCLUSIONS: Cancer incidence has remained relatively stable over time, but as shown elsewhere in sub-Saharan Africa the disease is a threat in The Gambia. The infection related cancers which are mostly preventable (HBV in men and HPV/HIV in women were the most common. At the moment the data is not enough to detect an effect of hepatitis B vaccination on liver cancer incidence in The Gambia. However, we observed that monitoring case occurrence through PBCR is a key public health pre-requisite for rational planning and implementation of targeted interventions for improving the health of the population.

  8. Hepatitis B vaccines: protective efficacy and therapeutic potential.

    Science.gov (United States)

    Michel, M-L; Tiollais, P

    2010-08-01

    Worldwide, two billion people have at some time been infected by hepatitis B virus, 370 millions suffer from chronic infection and around one million die each year from HBV-related liver diseases of which liver cancer is the ultimate stage. Vaccination is the measure that is most effective in reducing the global incidence of hepatitis B and hepatitis B vaccines have now been available for over 20 years. The first hepatitis B vaccine was prepared from inactivated hepatitis B surface antigen particles purified from plasma of asymptomatic carriers of hepatitis B virus. Knowledge of the structure and genomic organization of hepatitis B virus has led to development of the first DNA recombinant vaccine. In preventing hepatocellular carcinoma development, hepatitis B virus vaccines are considered as the first available cancer vaccine. HBV vaccines have recently taken on a new role as therapeutic vaccines as an attempt to cure or to control hepatitis B virus infection in persistently infected individuals. PMID:20382485

  9. A Diffusion-Based Approach to Geminate Recombination of Heme Proteins with Small Ligands

    CERN Document Server

    Starovoitov, V S

    2002-01-01

    A model of postphotodissociative monomolecular (geminate) recombination of heme proteins with small ligands (NO, O2 or CO) is represented. The non-exponential decay with time for the probability to find a heme in unbound state is interpreted in terms of diffusion-like migration of ligabs physics/0212040and between protein cavities. The temporal behavior for the probability is obtained from numerical simulation and specified by two parameters: the time \\tau_{reb} of heme-ligand rebinding for the ligand localized inside the heme pocket and the time \\tau_{esc} of ligand escape from the pocket. The model is applied in the analysis of available experimental data for geminate reoxygenation of human hemoglobin HbA. Our simulation is in good agreement with the measurements. The analysis shows that the variation in pH of the solution (6.0

  10. Multi-Homologous Recombination-Based Gene Manipulation in the Rice Pathogen Fusarium fujikuroi

    Directory of Open Access Journals (Sweden)

    In Sun Hwang

    2016-06-01

    Full Text Available Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1 , which is associated with fumonisin B1 biosynthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi.

  11. Peptide-Recombinant VP6 Protein Based Enzyme Immunoassay for the Detection of Group A Rotaviruses in Multiple Host Species

    Science.gov (United States)

    Sircar, Subhankar; Saurabh, Sharad; Gulati, Baldev R.; Singh, Neeraj; Singh, Arvind Kumar; Joshi, Vinay G.; Banyai, Krisztian; Dhama, Kuldeep

    2016-01-01

    We developed a novel enzyme immunoassay for the detection of group A rotavirus (RVA) antigen in fecal samples of multiple host species. The assay is based on the detection of conserved VP6 protein using anti-recombinant VP6 antibodies as capture antibodies and anti-multiple antigenic peptide (identified and constructed from highly immunodominant epitopes within VP6 protein) antibodies as detector antibodies. The clinical utility of the assay was evaluated using a panel of 914 diarrhoeic fecal samples from four different host species (bovine, porcine, poultry and human) collected from diverse geographical locations in India. Using VP6- based reverse transcription-polymerase chain reaction (RT-PCR) as the gold standard, we found that the diagnostic sensitivity (DSn) and specificity (DSp) of the new assay was high [bovine (DSn = 94.2% & DSp = 100%); porcine (DSn = 94.6% & DSp = 93.3%); poultry (DSn = 74.2% & DSp = 97.7%) and human (DSn = 82.1% & DSp = 98.7%)]. The concordance with RT-PCR was also high [weighted kappa (k) = 0.831–0.956 at 95% CI = 0.711–1.0] as compared to RNA-polyacrylamide gel electrophoresis (RNA-PAGE). The performance characteristics of the new immunoassay were comparable to those of the two commercially available ELISA kits. Our results suggest that this peptide-recombinant protein based assay may serve as a preliminary assay for epidemiological surveillance of RVA antigen and for evaluation of vaccine effectiveness especially in low and middle income settings. PMID:27391106

  12. Peptide-Recombinant VP6 Protein Based Enzyme Immunoassay for the Detection of Group A Rotaviruses in Multiple Host Species.

    Directory of Open Access Journals (Sweden)

    Naveen Kumar

    Full Text Available We developed a novel enzyme immunoassay for the detection of group A rotavirus (RVA antigen in fecal samples of multiple host species. The assay is based on the detection of conserved VP6 protein using anti-recombinant VP6 antibodies as capture antibodies and anti-multiple antigenic peptide (identified and constructed from highly immunodominant epitopes within VP6 protein antibodies as detector antibodies. The clinical utility of the assay was evaluated using a panel of 914 diarrhoeic fecal samples from four different host species (bovine, porcine, poultry and human collected from diverse geographical locations in India. Using VP6- based reverse transcription-polymerase chain reaction (RT-PCR as the gold standard, we found that the diagnostic sensitivity (DSn and specificity (DSp of the new assay was high [bovine (DSn = 94.2% & DSp = 100%; porcine (DSn = 94.6% & DSp = 93.3%; poultry (DSn = 74.2% & DSp = 97.7% and human (DSn = 82.1% & DSp = 98.7%]. The concordance with RT-PCR was also high [weighted kappa (k = 0.831-0.956 at 95% CI = 0.711-1.0] as compared to RNA-polyacrylamide gel electrophoresis (RNA-PAGE. The performance characteristics of the new immunoassay were comparable to those of the two commercially available ELISA kits. Our results suggest that this peptide-recombinant protein based assay may serve as a preliminary assay for epidemiological surveillance of RVA antigen and for evaluation of vaccine effectiveness especially in low and middle income settings.

  13. Expression of the classical swine fever E2 recombinant protein and examination of DNA-vaccine based on one subunit

    International Nuclear Information System (INIS)

    Full text: The aim of the work was to study the possibility of using the Classical Swine Fever Virus (CSFV) E2 (gp51-C55) recombinant protein expressed in E.coli in an immunologically active form and the creation of a DNA-vaccine model based on the gene. For minimizing the refolding problems of the recombinant protein one of two subunits of CSFV E2protein was chosen for production. This subunit has the domains A and D with the conservative epitopes some of which, in the domain A, induce synthesis of virus neutralizing antibodies. Viral RNA was isolated from the CSFV virulent strain Shi-Min, which was produced in a continuous swine kidney PK-15 cell culture (passage 37) Oligonucleotide primers were designed based on the Brescia (GeneBank, M31768) strain sequence Sn 2647-C2668 and Asn 3492-C3471. For additional RT-PCR controls the obtained fragment (846 n. p.) from isolates from CSF outbreak in Ukraine in 1994. It was shown that only the samples from the acute form of disease, wherever the amplicon sizes corresponded to the Shi-Min fragment, readily hybridized under this in stringency conditions. The subsequent sequence analysis of the fragment and the phylogenetic analyses with the use of sequences of various origin strains and isolates, placed the virus in group 1, subgroup 1.1, according to the classification suggested by D. Paton in 1995. For protein synthesis 3 constructions for plasmids expressing with the insert of CSFV E2 gene were created: CSFV-pET24a (+) -C as the individual form of the protein with the molecular mass (m.m.) 34 kD; CSFV-pGEX-2T - as fused with glutathion-S-tsansphesase, m.m. 59 kD and CSFV-pLY -C as fused with one subunit of γ-galactisidase, m.m. 107 kD. All 3 variants of the protein were synthesized as inclusion bodies with an expression level of 15-C20%. After optimizing the purification and refolding conditions, antigenic properties of the proteins were characterized in an indirect ELISA and immunoperoxidase test with homologous and

  14. Geno2pheno[HCV] – A Web-based Interpretation System to Support Hepatitis C Treatment Decisions in the Era of Direct-Acting Antiviral Agents

    Science.gov (United States)

    Kalaghatgi, Prabhav; Sikorski, Anna Maria; Knops, Elena; Rupp, Daniel; Sierra, Saleta; Heger, Eva; Neumann-Fraune, Maria; Beggel, Bastian; Walker, Andreas; Timm, Jörg; Walter, Hauke; Obermeier, Martin; Kaiser, Rolf; Bartenschlager, Ralf; Lengauer, Thomas

    2016-01-01

    The face of hepatitis C virus (HCV) therapy is changing dramatically. Direct-acting antiviral agents (DAAs) specifically targeting HCV proteins have been developed and entered clinical practice in 2011. However, despite high sustained viral response (SVR) rates of more than 90%, a fraction of patients do not eliminate the virus and in these cases treatment failure has been associated with the selection of drug resistance mutations (RAMs). RAMs may be prevalent prior to the start of treatment, or can be selected under therapy, and furthermore they can persist after cessation of treatment. Additionally, certain DAAs have been approved only for distinct HCV genotypes and may even have subtype specificity. Thus, sequence analysis before start of therapy is instrumental for managing DAA-based treatment strategies. We have created the interpretation system geno2pheno[HCV] (g2p[HCV]) to analyse HCV sequence data with respect to viral subtype and to predict drug resistance. Extensive reviewing and weighting of literature related to HCV drug resistance was performed to create a comprehensive list of drug resistance rules for inhibitors of the HCV protease in non-structural protein 3 (NS3-protease: Boceprevir, Paritaprevir, Simeprevir, Asunaprevir, Grazoprevir and Telaprevir), the NS5A replicase factor (Daclatasvir, Ledipasvir, Elbasvir and Ombitasvir), and the NS5B RNA-dependent RNA polymerase (Dasabuvir and Sofosbuvir). Upon submission of up to eight sequences, g2p[HCV] aligns the input sequences, identifies the genomic region(s), predicts the HCV geno- and subtypes, and generates for each DAA a drug resistance prediction report. g2p[HCV] offers easy-to-use and fast subtype and resistance analysis of HCV sequences, is continuously updated and freely accessible under http://hcv.geno2pheno.org/index.php. The system was partially validated with respect to the NS3-protease inhibitors Boceprevir, Telaprevir and Simeprevir by using data generated with recombinant, phenotypic

  15. Geno2pheno[HCV] - A Web-based Interpretation System to Support Hepatitis C Treatment Decisions in the Era of Direct-Acting Antiviral Agents.

    Science.gov (United States)

    Kalaghatgi, Prabhav; Sikorski, Anna Maria; Knops, Elena; Rupp, Daniel; Sierra, Saleta; Heger, Eva; Neumann-Fraune, Maria; Beggel, Bastian; Walker, Andreas; Timm, Jörg; Walter, Hauke; Obermeier, Martin; Kaiser, Rolf; Bartenschlager, Ralf; Lengauer, Thomas

    2016-01-01

    The face of hepatitis C virus (HCV) therapy is changing dramatically. Direct-acting antiviral agents (DAAs) specifically targeting HCV proteins have been developed and entered clinical practice in 2011. However, despite high sustained viral response (SVR) rates of more than 90%, a fraction of patients do not eliminate the virus and in these cases treatment failure has been associated with the selection of drug resistance mutations (RAMs). RAMs may be prevalent prior to the start of treatment, or can be selected under therapy, and furthermore they can persist after cessation of treatment. Additionally, certain DAAs have been approved only for distinct HCV genotypes and may even have subtype specificity. Thus, sequence analysis before start of therapy is instrumental for managing DAA-based treatment strategies. We have created the interpretation system geno2pheno[HCV] (g2p[HCV]) to analyse HCV sequence data with respect to viral subtype and to predict drug resistance. Extensive reviewing and weighting of literature related to HCV drug resistance was performed to create a comprehensive list of drug resistance rules for inhibitors of the HCV protease in non-structural protein 3 (NS3-protease: Boceprevir, Paritaprevir, Simeprevir, Asunaprevir, Grazoprevir and Telaprevir), the NS5A replicase factor (Daclatasvir, Ledipasvir, Elbasvir and Ombitasvir), and the NS5B RNA-dependent RNA polymerase (Dasabuvir and Sofosbuvir). Upon submission of up to eight sequences, g2p[HCV] aligns the input sequences, identifies the genomic region(s), predicts the HCV geno- and subtypes, and generates for each DAA a drug resistance prediction report. g2p[HCV] offers easy-to-use and fast subtype and resistance analysis of HCV sequences, is continuously updated and freely accessible under http://hcv.geno2pheno.org/index.php. The system was partially validated with respect to the NS3-protease inhibitors Boceprevir, Telaprevir and Simeprevir by using data generated with recombinant, phenotypic

  16. Geno2pheno[HCV] - A Web-based Interpretation System to Support Hepatitis C Treatment Decisions in the Era of Direct-Acting Antiviral Agents.

    Directory of Open Access Journals (Sweden)

    Prabhav Kalaghatgi

    Full Text Available The face of hepatitis C virus (HCV therapy is changing dramatically. Direct-acting antiviral agents (DAAs specifically targeting HCV proteins have been developed and entered clinical practice in 2011. However, despite high sustained viral response (SVR rates of more than 90%, a fraction of patients do not eliminate the virus and in these cases treatment failure has been associated with the selection of drug resistance mutations (RAMs. RAMs may be prevalent prior to the start of treatment, or can be selected under therapy, and furthermore they can persist after cessation of treatment. Additionally, certain DAAs have been approved only for distinct HCV genotypes and may even have subtype specificity. Thus, sequence analysis before start of therapy is instrumental for managing DAA-based treatment strategies. We have created the interpretation system geno2pheno[HCV] (g2p[HCV] to analyse HCV sequence data with respect to viral subtype and to predict drug resistance. Extensive reviewing and weighting of literature related to HCV drug resistance was performed to create a comprehensive list of drug resistance rules for inhibitors of the HCV protease in non-structural protein 3 (NS3-protease: Boceprevir, Paritaprevir, Simeprevir, Asunaprevir, Grazoprevir and Telaprevir, the NS5A replicase factor (Daclatasvir, Ledipasvir, Elbasvir and Ombitasvir, and the NS5B RNA-dependent RNA polymerase (Dasabuvir and Sofosbuvir. Upon submission of up to eight sequences, g2p[HCV] aligns the input sequences, identifies the genomic region(s, predicts the HCV geno- and subtypes, and generates for each DAA a drug resistance prediction report. g2p[HCV] offers easy-to-use and fast subtype and resistance analysis of HCV sequences, is continuously updated and freely accessible under http://hcv.geno2pheno.org/index.php. The system was partially validated with respect to the NS3-protease inhibitors Boceprevir, Telaprevir and Simeprevir by using data generated with recombinant

  17. Immunoprophylaxis to limit a hepatitis B epidemic among women undergoing in vitro fertilization

    OpenAIRE

    Grosheide, Pia Maria; Osand, H.; Schalm, Solko; Heijtink, R A

    1991-01-01

    markdownabstractAbstract Women (175) who participated in an in vitro fertilization (IVF) programme were possibly exposed to hepatitis B virus. Later it became evident that 79 women had a hepatitis B infection, 49 were exposed but not infected and 47 were not exposed. Hepatitis B immunoglobulin (HBIg) and recombinant hepatitis B vaccine (HBvaxDNA) were offered to all women and partners except for those with established hepatitis B. Women were given an ‘intensive’ schedule of HBIg (0, 1 months)...

  18. A novel hepatitis C virus genotyping method based on liquid microarray.

    Directory of Open Access Journals (Sweden)

    Cesar A B Duarte

    Full Text Available The strategy used to treat HCV infection depends on the genotype involved. An accurate and reliable genotyping method is therefore of paramount importance. We describe here, for the first time, the use of a liquid microarray for HCV genotyping. This liquid microarray is based on the 5'UTR - the most highly conserved region of HCV - and the variable region NS5B sequence. The simultaneous genotyping of two regions can be used to confirm findings and should detect inter-genotypic recombination. Plasma samples from 78 patients infected with viruses with genotypes and subtypes determined in the Versant™ HCV Genotype Assay LiPA (version I; Siemens Medical Solutions, Diagnostics Division, Fernwald, Germany were tested with our new liquid microarray method. This method successfully determined the genotypes of 74 of the 78 samples previously genotyped in the Versant™ HCV Genotype Assay LiPA (74/78, 95%. The concordance between the two methods was 100% for genotype determination (74/74. At the subtype level, all 3a and 2b samples gave identical results with both methods (17/17 and 7/7, respectively. Two 2c samples were correctly identified by microarray, but could only be determined to the genotype level with the Versant™ HCV assay. Genotype "1" subtypes (1a and 1b were correctly identified by the Versant™ HCV assay and the microarray in 68% and 40% of cases, respectively. No genotype discordance was found for any sample. HCV was successfully genotyped with both methods, and this is of prime importance for treatment planning. Liquid microarray assays may therefore be added to the list of methods suitable for HCV genotyping. It provides comparable results and may readily be adapted for the detection of other viruses frequently co-infecting HCV patients. Liquid array technology is thus a reliable and promising platform for HCV genotyping.

  19. Hepatitis Vaccines.

    Science.gov (United States)

    Ogholikhan, Sina; Schwarz, Kathleen B

    2016-01-01

    Viral hepatitis is a serious health problem all over the world. However, the reduction of the morbidity and mortality due to vaccinations against hepatitis A and hepatitis B has been a major component in the overall reduction in vaccine preventable diseases. We will discuss the epidemiology, vaccine development, and post-vaccination effects of the hepatitis A and B virus. In addition, we discuss attempts to provide hepatitis D vaccine for the 350 million individuals infected with hepatitis B globally. Given the lack of a hepatitis C vaccine, the many challenges facing the production of a hepatitis C vaccine will be shown, along with current and former vaccination trials. As there is no current FDA-approved hepatitis E vaccine, we will present vaccination data that is available in the rest of the world. Finally, we will discuss the existing challenges and questions facing future endeavors for each of the hepatitis viruses, with efforts continuing to focus on dramatically reducing the morbidity and mortality associated with these serious infections of the liver. PMID:26978406

  20. Hepatitis Vaccines

    Directory of Open Access Journals (Sweden)

    Sina Ogholikhan

    2016-03-01

    Full Text Available Viral hepatitis is a serious health problem all over the world. However, the reduction of the morbidity and mortality due to vaccinations against hepatitis A and hepatitis B has been a major component in the overall reduction in vaccine preventable diseases. We will discuss the epidemiology, vaccine development, and post-vaccination effects of the hepatitis A and B virus. In addition, we discuss attempts to provide hepatitis D vaccine for the 350 million individuals infected with hepatitis B globally. Given the lack of a hepatitis C vaccine, the many challenges facing the production of a hepatitis C vaccine will be shown, along with current and former vaccination trials. As there is no current FDA-approved hepatitis E vaccine, we will present vaccination data that is available in the rest of the world. Finally, we will discuss the existing challenges and questions facing future endeavors for each of the hepatitis viruses, with efforts continuing to focus on dramatically reducing the morbidity and mortality associated with these serious infections of the liver.

  1. Evaluation of hyaluronic acid-based combination adjuvant containing monophosphoryl lipid A and aluminum salt for hepatitis B vaccine.

    Science.gov (United States)

    Moon, Se-hee; Shin, Eui-Cheol; Noh, Young-Woock; Lim, Yong Taik

    2015-09-11

    Here, monophosphoryl lipid A (MPLA) and aluminum salt (Alum) were introduced into a hyaluronic acid (HA)-based combination vaccine adjuvant for hepatitis B vaccine (HBV). Although Alum is a well-known hepatitis B vaccine adjuvant that induces an enhanced humoral immune response, it cannot induce the cellular immune responses. On the other hand, MPLA has been generally reported to promote IFN-γ production via antigen-specific CD4(+) T cells, but it is not water soluble as a result of its long hydrophobic alkyl chains. To this end, water insoluble MPLA could be solubilized in an aqueous solution with the help of HA, which contains many carboxyl and hydroxyl groups that can be used to attach to the hydroxyl head groups of MPLA via hydrogen bonds. Three groups of mice were treated with either hepatitis B surface antigen (HBsAg) alone, HBsAg_Alum complex, or HBsAg_Alum_MPLA/HA complex. The group immunized with the HBsAg_Alum_MPLA/HA complex exhibited a high increase in cellular immune response as well as in humoral immune response relative to the other two groups. The antibody, cytokine and T cell levels were most elevated in the group of mice immunized with HBsAg_Alum_MPLA/HA complex, even at a 1μg/mice dose, and the magnitude was still maintained even after 8 weeks. Specifically, the antibody value was 120 times larger in mice vaccinated with HBsAg_Alum_MPLA/HA complex than in mice vaccinated with HBsAg_Alum complex designed similar to commercially available hepatitis B vaccine, Engerix B. The cytokine and T cell proliferation levels were 2 times and 6 times larger in mice adjuvanted with HBsAg_Alum_MPLA/HA complex than in those vaccinated with HBsAg_Alum. The results therefore indicate that incorporating MPLA and Alum with HA can be a potent strategy to increase both the magnitude and the persistence of HBsAg-specific immune responses to protect hosts against hepatitis B virus infection. PMID:26271830

  2. Development of recombinant collagen-peptide-based vehicles for delivery of adipose-derived stromal cells.

    Science.gov (United States)

    Parvizi, Mojtaba; Plantinga, Josée A; van Speuwel-Goossens, Carolina A F M; van Dongen, Elisabeth M W M; Kluijtmans, Sebastiaan G J M; Harmsen, Martin C

    2016-02-01

    Stem cell therapy is a promising approach for repair, remodeling and even regenerate tissue of otherwise irreparable damage, such as after myocardial infarction (aMI). A severe limitation of cardiac stem cell therapy is the generally poor retention of administered cells in the target tissue. In tissue repair the main mode of action of adipose tissue-derived stem cells (ADSC) is the production of various growth factors, cytokines, anti-inflammatory and anti-apoptotic factors that together augment repair, remodeling, and regeneration. In this experiment, we used recombinant collagen peptide (RCP) with additional integrin-binding motives and different crosslinkers. Formulated as 50-100 μm microspheres with bound ADSC, we hypothesized that this would improve ADSC retention and function. Crosslinking was performed with chemical crosslinkers (EDC and HMDIC) at high and low concentrations or by thermal treatment (DHT). ADSC adhesion, proliferation, apoptosis/necrosis, and gene expressions in two-dimensional and three-dimensional were analyzed. In addition, the effect of ADSC conditioned medium (ADSC-CM) on proapoptotic/sprouting HUVEC was examined. Our results show that all materials support cell adhesion in short time point, however, EDC-High crosslinker induced ADSC apoptosis/necrosis. Gene expression results revealed lower expression of proinflammatory genes in chemical crosslinked materials, despite EDC-High the proinflammatory genes expressions were similar or higher than TCPS. In addition, cultured ADSC on DHT crosslinked RCP showed a proinflammatory phenotype compared to TCPS. Sprouting assay results confirmed the protective effect of ADSC-CM derived from TCPS and HMDIC-High crosslinked RCP proapoptotic HUVEC. We conclude that ADSC adhere to the materials and maintain their therapeutic profile.

  3. A population-based study examining hepatitis B virus infection and immunization rates in Northwest China.

    Directory of Open Access Journals (Sweden)

    Zhaohua Ji

    Full Text Available BACKGROUND AND AIM: Current baseline data regarding the prevalence of hepatitis B virus (HBV infections and the immune status in hyperendemic areas is necessary in evaluating the effectiveness of ongoing HBV prevention and control programs in northwest China. This study aims to determine the prevalence of chronic HBV infections, past exposure rates, and immune response profiles in Wuwei City, northwest China in 2010. METHODS: Cross-sectional household survey representative of the Wuwei City population. 28,579 participants were interviewed in the seroepidemiological survey ≥1 year of age. House to house screening was conducted using a standard questionnaire. All serum samples were screened by enzyme-linked immunoassays for the presence of hepatitis B surface antigen, antibodies against HBV surface antigen, and antibodies to the hepatitis B core antigen. RESULTS: Among individuals ≥1 year of age, 7.2% (95%CI: 6.3-8.1% had chronic HBV infections, 43.9% (CI: 40.4-47.4% had been exposed to HBV, and 23.49% (CI: 21.6-25.3% had vaccine-induced immunity. Multi-factor weighted logistic regression analysis showed that having household contact with HBV carriers (OR = 2.6, 95%CI: 2.3-3.0 and beauty treatments in public places (OR = 1.2, 95%CI: 1.1-1.3 were the risk factors of HBV infection in whole population. Having household contact with HBV carriers (OR = 3.8, 95% CI: 2.2-6.5 and lack of hepatitis vaccination (OR = 2.0, 95% CI: 1.4-3.3 were the risk factors for HBV infection in children aged 1-14 years. CONCLUSIONS: Hepatitis B infection remains a serious public health problem in northwest China. Having household contact with HBV carriers and beauty treatments in public places represented HBV infection risk factors. Hepatitis B vaccine immunization strategies need further improvement, particularly by targeting the immunization of rural migrant workers.

  4. Positive Impact of a Shelter-based Hepatitis B Vaccine Program in Homeless Baltimore Children and Adolescents

    OpenAIRE

    Schwarz, Kathleen; Garrett, Beth; Lee, Jennifer; Thompson, Douglas; Thiel, Thelma; Alter, Miriam J.; Strathdee, Stephanie

    2008-01-01

    Homeless youth are at increased risk for hepatitis B virus (HBV) infection and HBV vaccine coverage is poor in this group. The purpose of our study was to determine if a shelter-based HBV vaccine program in children and adolescents 2–18 years of age with a randomized controlled trial using a culturally appropriate HBV video could increase HBV vaccine coverage rates. Subjects were randomized to an 8 min HBV video or a control, smoking prevention video. Before exposure to the videos, HBV knowle...

  5. The key role for local base order in the generation of multiple forms of China HIV-1 B'/C intersubtype recombinants

    Directory of Open Access Journals (Sweden)

    Wei Ji-Fu

    2005-10-01

    Full Text Available Abstract Background HIV-1 is a retrovirus with high rate of recombination. Increasing experimental studies in vitro indicated that local hairpin structure of RNA was associated with recombination by favoring RT pausing and promoting strand transfer. A method to estimate the potential to form stem-loop structure by calculating the folding of randomized sequence difference (FORS-D has been used to investigate the relationship between secondary structure and evolutionary pressure in some genome. It showed that gene regions under strong positive "Darwinian" selection were associated with positive FORS-D values. In the present study, the sequences of HIV-1 subtypes B' and C, both of which represent the parent strains of CRF07_BC, CRF08_BC and China URFs, were selected to investigate the relationship between natural recombination and secondary structure by calculating the FORS-D values. Results The apparent higher negative FORS-D value region appeared in the gag-pol gene region (nucleotide 0–3000 of HIV-1 subtypes B' and C. Thirteen (86.7 % of 15 mosaic fragments and 17 (81 % of 21 recombination breakpoints occurred in this higher negative FORS-D region. This strongly suggested that natural recombination did not occur randomly throughout the HIV genome, and that there might be preferred (or hot regions or sites for recombination. The FORS-D analysis of breakpoints showed that most breakpoints of recombinants were located in regions with higher negative FORS-D values (P = 0.0053, and appeared to have a higher negative average FORS-D value than the whole genome (P = 0.0007. The regression analysis also indicated that FORS-D values correlated negatively with breakpoint overlap. Conclusion High negative FORS-D values represent high, base order determined stem-loop potentials and influence mainly the formation of stem-loop structures. Therefore, the present results suggested for the first time that occurrence of natural recombination was associated with

  6. Feature Hepatitis: Hepatitis Can Strike Anyone

    Science.gov (United States)

    ... Navigation Bar Home Current Issue Past Issues Feature Hepatitis Hepatitis Can Strike Anyone Past Issues / Spring 2009 Table ... from all walks of life are affected by hepatitis, especially hepatitis C, the most common form of ...

  7. Identification of Drought Tolerant Mechanisms in Maize Seedlings Based on Transcriptome Analysis of Recombination Inbred Lines.

    Science.gov (United States)

    Min, Haowei; Chen, Chengxuan; Wei, Shaowei; Shang, Xiaoling; Sun, Meiyun; Xia, Ran; Liu, Xiangguo; Hao, Dongyun; Chen, Huabang; Xie, Qi

    2016-01-01

    Zea mays is an important crop that is sensitive to drought stress, but survival rates and growth status remain strong in some drought-tolerant lines under stress conditions. Under drought conditions, many biological processes, such as photosynthesis, carbohydrate metabolism and energy metabolism, are suppressed, while little is known about how the transcripts of genes respond to drought stress in the genome-wide rang in the seedling stage. In our study, the transcriptome profiles of two maize recombination inbred lines (drought-tolerant RIL70 and drought-sensitive RIL93) were analyzed at different drought stages to elucidate the dynamic mechanisms underlying drought tolerance in maize seedlings during drought conditions. Different numbers of differentially expressed genes presented in the different stages of drought stress in the two RILs, for the numbers of RIL93 vs. RIL70 were: 9 vs. 358, 477 vs. 103, and 5207 vs. 152 respectively in DT1, DT2, and DT5. Gene Ontology enrichment analysis revealed that in the initial drought-stressed stage, the primary differentially expressed genes involved in cell wall biosynthesis and transmembrane transport biological processes were overrepresented in RIL70 compared to RIL93. On the contrary, differentially expressed genes profiles presented at 2 and 5 day-treatments, the primary differentially expressed genes involved in response to stress, protein folding, oxidation-reduction, photosynthesis and carbohydrate metabolism, were overrepresented in RIL93 compared to RIL70. In addition, the transcription of genes encoding key members of the cell cycle and cell division processes were blocked, but ABA- and programmed cell death-related processes responded positively in RIL93. In contrast, the expression of cell cycle genes, ABA- and programmed cell death-related genes was relatively stable in RIL70. The results we obtained supported the working hypothesis that signaling events associated with turgor homeostasis, as established by

  8. Discrimination Based on Health Grounds : Case Study: Hepatitis B Virus Discrimination in China Labour Employment

    OpenAIRE

    2006-01-01

    Nowadays, due to the high prevalence of hepatitis B in China, millions of carriers are faced with discrimination when they come to work, study, health care or even marriage. The same situation also happens to those physically disadvantageous people especially in the access to employment. Employment discrimination detracts from the principle of equality and directly impairs social justice and human dignity. Series of institutional responses are needed to effectively prevent employment discrimi...

  9. HPMA Polymer-based Site-specific Delivery of Oligonucleotides to Hepatic Stellate Cells

    OpenAIRE

    Yang, Ningning; Ye, Zhaoyang; Li, Feng; Mahato, Ram I.

    2009-01-01

    The objective was to determine whether bioconjugation of type I collagen specific triplex forming oligonucleotide (TFO) to N-(2-hydroxypropyl) methacrylamide (HPMA) containing tetrapeptide Gly-Phe-Leu-Gly (GFLG) and mannose 6-phosphate (M6P) can provide their targeted delivery to hepatic stellate cells (HSCs). Following bioconjugation, M6P-GFLG-HPMA-GFLG-32P-TFO was characterized by PAGE, HPLC and GPC, and then its biodistribution was determined. TFO was dissociated from the conjugate when in...

  10. Viscoelasticity-based magnetic resonance elastography for the assessment of liver fibrosis in hepatitis C patients after liver transplantation

    Energy Technology Data Exchange (ETDEWEB)

    Kamphues, C.; Bova, R.; Yahyazadeh, A.; Bahra, M.; Neuhaus, P. [Charite, Campus Virchow Klinikum, Berlin (Germany). Klinik fuer Allgemein-, Viszeral- und Transplantationschirurgie; Klatt, D.; Braun, J.; Sack, I.; Asbach, P. [Charite - Universitaetsmedizin, Berlin (Germany). Klinik fuer Radiologie; Klauschen, F. [Charite - Universitaetsmedizin, Berlin (Germany). Inst. fuer Pathologie

    2012-11-15

    Purpose: Despite advantages in antiviral therapy of hepatitis C (HCV) in recent years, progressing liver fibrosis remains a major problem for patients suffering from hepatitis C after liver transplantation. Therefore, effective non-invasive methods for the assessment of liver fibrosis are needed in order to guide treatment decisions and predict prognosis in these patients. The aim of this study was to prospectively assess the diagnostic accuracy of viscoelasticity-based magnetic resonance (MR) elastography for the assessment of liver fibrosis in HCV patients after liver transplantation. Materials and Methods: After IRB approval, a total of 25 patients, who had received a liver graft due to chronic hepatitis C underwent both liver biopsy and MR elastography. Two viscoelastic constants, the shear elasticity {mu} and the powerlaw exponent {alpha} were calculated by fitting the frequency function of the complex shear modulus with the viscoelastic springpot-model. Results: A strong positive correlation between shear elasticity {mu} and the stage of fibrosis could be found (R = 0.486, p = 0.0136). The area under the receiver operating curve (AUROC) of MR elastography based on {mu} for diagnosis of severe fibrosis (F {>=} 3) was 0.87 and 0.65 for diagnosis of significant fibrosis (F {>=} 2). The powerlaw exponent {alpha} did not correlate with the stage of fibrosis. Conclusion: MR elastography represents a promising non-invasive procedure for the assessment of higher grades of fibrosis in HCV patients after liver transplantation. The poor correlation for lower grades of fibrosis suggests unknown mechanical interactions in the transplanted liver. (orig.)

  11. Viscoelasticity-based magnetic resonance elastography for the assessment of liver fibrosis in hepatitis C patients after liver transplantation

    International Nuclear Information System (INIS)

    Purpose: Despite advantages in antiviral therapy of hepatitis C (HCV) in recent years, progressing liver fibrosis remains a major problem for patients suffering from hepatitis C after liver transplantation. Therefore, effective non-invasive methods for the assessment of liver fibrosis are needed in order to guide treatment decisions and predict prognosis in these patients. The aim of this study was to prospectively assess the diagnostic accuracy of viscoelasticity-based magnetic resonance (MR) elastography for the assessment of liver fibrosis in HCV patients after liver transplantation. Materials and Methods: After IRB approval, a total of 25 patients, who had received a liver graft due to chronic hepatitis C underwent both liver biopsy and MR elastography. Two viscoelastic constants, the shear elasticity μ and the powerlaw exponent α were calculated by fitting the frequency function of the complex shear modulus with the viscoelastic springpot-model. Results: A strong positive correlation between shear elasticity μ and the stage of fibrosis could be found (R = 0.486, p = 0.0136). The area under the receiver operating curve (AUROC) of MR elastography based on μ for diagnosis of severe fibrosis (F ≥ 3) was 0.87 and 0.65 for diagnosis of significant fibrosis (F ≥ 2). The powerlaw exponent α did not correlate with the stage of fibrosis. Conclusion: MR elastography represents a promising non-invasive procedure for the assessment of higher grades of fibrosis in HCV patients after liver transplantation. The poor correlation for lower grades of fibrosis suggests unknown mechanical interactions in the transplanted liver. (orig.)

  12. Evidence-based consensus on the diagnosis, prevention andmanagement of hepatitis C virus disease

    Institute of Scientific and Technical Information of China (English)

    Mahrukh Akbar Shaheen; Muhammad Idrees

    2015-01-01

    Hepatitis C virus (HCV) is a potent human pathogenand is one of the main causes of chronic hepatitis roundthe world. The present review describes the evidencebasedconsensus on the diagnosis, prevention andmanagement of HCV disease. Various techniques, for thedetection of anti-HCV immunoglobulin G immunoassays,detection of HCV RNA by identifying virus-specificmolecules nucleic acid testings, recognition of coreantigen for diagnosis of HCV, quantitative antigenassay, have been used to detect HCV RNA and coreantigen. Advanced technologies such as nanoparticlebaseddiagnostic assays, loop-mediated isothermalamplification and aptamers and Ortho trak-C assayhave also come to the front that provides best detectionresults with greater ease and specificity for detectionof HCV. It is of immense importance to prevent thisinfection especially among the sexual partners, injectingdrug users, mother-to-infant transmission of HCV,household contact, healthcare workers and people whoget tattoos and piercing on their skin. Management of thisinfection is intended to eradicate it out of the body ofpatients. Management includes examining the treatment(efficacy and protection), assessment of hepatic conditionbefore commencing therapy, controlling theparameters upon which dual and triple therapies work,monitoring the body after treatment and adjusting theco-factors. Examining the treatment in some specialgroups of people (HIV/HCV co-infected, hemodialysispatients, renal transplanted patients).

  13. Hepatitis (For Parents)

    Science.gov (United States)

    ... Tropical Delight: Melon Smoothie Pregnant? Your Baby's Growth Hepatitis KidsHealth > For Parents > Hepatitis Print A A A ... to Call the Doctor en español Hepatitis About Hepatitis The word hepatitis simply means an inflammation of ...

  14. An in vitro recombination-based reverse genetic system for rapid mutagenesis of structural genes of the Japanese encephalitis virus

    Institute of Scientific and Technical Information of China (English)

    Ruikun; Du; Manli; Wang; Zhihong; Hu; Hualin; Wang; Fei; Deng

    2015-01-01

    Japanese encephalitis virus(JEV) is one of the most common pathogens of severe viral encephalitis, which is a severe threat to human health. Despite instability of the JEV genome in bacteria, many strategies have been developed to establish molecular clone systems of JEV, providing convenient tools for studying the virus life cycle and virus–host interactions. In this study, we adapted an In-Fusion enzyme-based in vitro recombination method to construct a reverse genetic system of JEV, thereby providing a rapid approach to introduce mutations into the structural genes. A truncated genome without the structural genes was constructed as the backbone, and the complementary segment containing the structural genes was recombined in vitro, which was then transfected directly into virus-permissive cells. The progeny of the infectious virus was successfully detected in the supernatant of the transfected cells, and showed an identical phenotype to its parental virus. To provide a proof-of-principle, the 12 conserved cysteine residues in the envelope(E) protein of JEV were respectively mutated using this approach, and all mutations resulted in a complete failure to generate infectious virus. However, a leucine-tophenylanine mutation at amino acid 107 of the E protein did not interfere with the production of the infectious virus. These results suggested that all 12 cysteines in the E protein are essential for the JEV life cycle. In summary, a novel reverse genetic system of JEV was established for rapidly introducing mutations into structural genes, which will serve as a useful tool for functional studies.

  15. Phase I Clinical Trial of a Recombinant Blood Stage Vaccine Candidate for Plasmodium falciparum Malaria Based on MSP1 and EBA175.

    Directory of Open Access Journals (Sweden)

    Chetan E Chitnis

    Full Text Available A phase I randomised, controlled, single blind, dose escalation trial was conducted to evaluate safety and immunogenicity of JAIVAC-1, a recombinant blood stage vaccine candidate against Plasmodium falciparum malaria, composed of a physical mixture of two recombinant proteins, PfMSP-1(19, the 19 kD conserved, C-terminal region of PfMSP-1 and PfF2 the receptor-binding F2 domain of EBA175.Healthy malaria naïve Indian male subjects aged 18-45 years were recruited from the volunteer database of study site. Fifteen subjects in each cohort, randomised in a ratio of 2:1 and meeting the protocol specific eligibility criteria, were vaccinated either with three doses (10 μg, 25 μg and 50 μg of each antigen of JAIVAC-1 formulated with adjuvant Montanide ISA 720 or with standard dosage of Hepatitis B vaccine. Each subject received the assigned vaccine in the deltoid muscle of the upper arms on Day 0, Day 28 and Day 180.JAIVAC-1 was well tolerated and no serious adverse event was observed. All JAIVAC-1 subjects sero-converted for PfF2 but elicited poor immune response to PfMSP-1(19. Dose-response relationship was observed between vaccine dose of PfF2 and antibody response. The antibodies against PfF2 were predominantly of IgG1 and IgG3 isotype. Sera from JAIVAC-1 subjects reacted with late schizonts in a punctate pattern in immunofluorescence assays. Purified IgG from JAIVAC-1 sera displayed significant growth inhibitory activity against Plasmodium falciparum CAMP strain.Antigen PfF2 should be retained as a component of a recombinant malaria vaccine but PfMSP-1(19 construct needs to be optimised to improve its immunogenicity.Clinical Trial Registry, India CTRI/2010/091/000301.

  16. Phase I Clinical Trial of a Recombinant Blood Stage Vaccine Candidate for Plasmodium falciparum Malaria Based on MSP1 and EBA175

    Science.gov (United States)

    Chitnis, Chetan E.; Mukherjee, Paushali; Mehta, Shantanu; Yazdani, Syed Shams; Dhawan, Shikha; Shakri, Ahmad Rushdi; Bharadwaj, Rukmini; Gupta, Puneet Kumar; Hans, Dhiraj; Mazumdar, Suman; Singh, Bijender; Kumar, Sanjeev; Pandey, Gaurav; Parulekar, Varsha; Imbault, Nathalie; Shivyogi, Preethi; Godbole, Girish; Mohan, Krishna; Leroy, Odile; Singh, Kavita; Chauhan, Virander S.

    2015-01-01

    Background A phase I randomised, controlled, single blind, dose escalation trial was conducted to evaluate safety and immunogenicity of JAIVAC-1, a recombinant blood stage vaccine candidate against Plasmodium falciparum malaria, composed of a physical mixture of two recombinant proteins, PfMSP-119, the 19 kD conserved, C-terminal region of PfMSP-1 and PfF2 the receptor-binding F2 domain of EBA175. Method Healthy malaria naïve Indian male subjects aged 18–45 years were recruited from the volunteer database of study site. Fifteen subjects in each cohort, randomised in a ratio of 2:1 and meeting the protocol specific eligibility criteria, were vaccinated either with three doses (10μg, 25μg and 50μg of each antigen) of JAIVAC-1 formulated with adjuvant Montanide ISA 720 or with standard dosage of Hepatitis B vaccine. Each subject received the assigned vaccine in the deltoid muscle of the upper arms on Day 0, Day 28 and Day 180. Results JAIVAC-1 was well tolerated and no serious adverse event was observed. All JAIVAC-1 subjects sero-converted for PfF2 but elicited poor immune response to PfMSP-119. Dose-response relationship was observed between vaccine dose of PfF2 and antibody response. The antibodies against PfF2 were predominantly of IgG1 and IgG3 isotype. Sera from JAIVAC-1 subjects reacted with late schizonts in a punctate pattern in immunofluorescence assays. Purified IgG from JAIVAC-1 sera displayed significant growth inhibitory activity against Plasmodium falciparum CAMP strain. Conclusion Antigen PfF2 should be retained as a component of a recombinant malaria vaccine but PfMSP-119 construct needs to be optimised to improve its immunogenicity. Trial Registration Clinical Trial Registry, India CTRI/2010/091/000301 PMID:25927360

  17. Biodegradable Microspheres as Hepatitis B Vaccine Delivery Systems

    Institute of Scientific and Technical Information of China (English)

    杨春; 贾文祥; 陈恬; 曾蔚; 杨远; 杨发龙; 谢轶; 杨维清; 周绍兵; 李孝红

    2003-01-01

    In order to investigate the immtmogenicity of the controlled-release microencapsulated hepatitis B vaccine in mice, polyethylene glycol-poly-dl-lactide (PELA) microspheres with entrapped HSsAg were prepared by double emulsion W/O/W based on solvent extraction methods. BALB/c mice were immunized with the encapsulated vaccine by oral feeding or injection. Blood samples were collected at 8th, 10th, 14th and 24th weeks, respectively, and the levels of antibody response were detected by EI.ISA. It was found that the scanning electron microscopy showed the prepared microspheres had smoothand spherical surface, suitable for vaccine delivery. Two groups of mice orally fed with the encapsulated or conventional recombinant vaccines, respectively, there sere showed no obvious difference in the IgG levels. At 14th week, the group injected with a single dose of encapsulated vaccine had a similar level of IgG response to the group injected with two doses of the recombination vaccine. At 24th week, the IgG levels of the group injected with two doses of encapsulated vaccine were higher than those of the group injected with two doses of the recombination vaccine. It concludes that Controlled-release microencapsulated hepatitis B vaccine possesses the feature of slowly releasing in v/vo and long times immtmogenicity.

  18. Pulmonary and cutaneous vasculitis following hepatitis B vaccination

    OpenAIRE

    Allen, Martin B; Cockwell, Paul; Page, Richard L.

    1993-01-01

    The case history is presented of a previously healthy non-atopic woman who developed cutaneous vasculitis, confirmed by biopsy, and pulmonary problems after inoculation with recombinant hepatitis B vaccine.

  19. 重组人干扰素联合阿德福韦酯治疗慢性乙型肝炎的疗效观察%THERAPEUTIC EFFECT OF RECOMBINANT HUMAN INTERFERON COMBINED WITH ADEFOVIR DIPIVOXIL IN THE TREATMENT OF CHRONIC HEPATITIS B

    Institute of Scientific and Technical Information of China (English)

    马红星

    2014-01-01

    目的:探讨重组干扰素联合阿德福韦酯治疗慢性乙型肝炎的临床疗效。方法将2011年12月-2012年12月72例慢性乙肝患者随机分成二组:对照组36例单纯采用重组人干扰素α-2b治疗;治疗组36例采用重组人干扰素α-2b+阿德福韦酯治疗;比较二组临床疗效、乙肝病毒脱氧核糖核酸(HBV DNA)转阴率、乙肝病毒e抗原(HBeAg)转阴率、HBeAg血清转换率和患者肝功能指标水平的变化情况。结果治疗组的总有效率明显高于对照组(P<0.05);治疗组治疗后的 HBV DNA转阴率、HBeAg转阴率、HBeAg血清转换率均显著高于对照组(P<0.05);治疗组患者治疗后的丙氨酸转氨酶(ALT )、冬氨酸转氨酶(AST ),与对照组差异无统计学意义。结论重组人干扰素联合阿德福韦酯治疗慢性乙型肝炎,提高乙肝病毒标志物转阴率,改善肝功能水平。%Objective To evaluate the clinical efficacy of recombinant interferon in combination with adefo-vir dipivoxil in the treatment of chronic hepatitis B .Methods Totally 72 cases of chronic hepatitis B pa-tients in author's hospital between December 2011 and December 2012 were randomly divided into two groups ,36 cases in each group .The control group was treated with only recombinant human interferon al-pha -2b;while the treatment group was treated with recombinant human interferon alpha-2b and adefo-vir dipivoxil .The clinical efficacy two groups was compared on HBV -DNA negative rate ,hepatitis B vi-rus e antigen(HBeAg) negative rate ,changes of HBeAg seroconversion rates and liver function indexes . Results The total effective rate in treatment group was significantly higher than the control group (P<0 .05);After treatment ,HBV -DNA negative conversion rate ,HBeAg negative conversion rate ,HBeAg seroconversion rates were significantly higher than the control groups (P<0 .05);There was no significant difference between the control group

  20. Experience with hepatitis A and B vaccines.

    Science.gov (United States)

    Davis, Jeffrey P

    2005-10-01

    The lengthy history of efforts to understand the pathogenesis and means of preventing and controlling both hepatitis A and B is noteworthy for many exceptional scientific achievements. Among these are the development of vaccines to prevent the spread of infection through induction of active immunity to hepatitis A virus (HAV) and hepatitis B virus (HBV). The first plasma-derived hepatitis B vaccine was licensed in the United States in 1981 and was replaced by recombinant hepatitis B vaccines in 1986 and 1989. Vaccines to prevent HAV infection were licensed in the United States in 1995 and 1996. Subsequently, combination vaccines that included both hepatitis A and B vaccine components, or the hepatitis B component in combination with other commonly administered vaccines, were licensed in the United States. Despite significant reductions in hepatitis-related morbidity and mortality that have resulted from widespread use of these vaccines, vaccine-preventable morbidity and mortality still occur. The purposes of this article are to review clinical trial and other experience with hepatitis A and B vaccines in healthy individuals as well as in those with chronic liver disease, infected with the human immunodeficiency virus, or requiring hemodialysis; describe the impact that these vaccines and national recommendations for vaccination have had on reducing the incidence of HAV and HBV infection; and recommend expansion of these recommendations to include universal vaccination of adults as a means of further reducing the burden of viral hepatitis.

  1. Autoimmune hepatitis triggered by acute hepatitis A

    Institute of Scientific and Technical Information of China (English)

    Hiroto Tanaka; Hiroto Tujioka; Hiroki Ueda; Hiroko Hamagami; Youhei Kida; Masakazu Ichinose

    2005-01-01

    The patient was a 57-year-old woman presenting with jaundice as the chief complaint. She began vomiting on July 10, 2003.Jaundice was noted and admitted to our hospital for thorough testing. Tests on admission indicated severe hepatitis, based on: aspartate aminotransferase (AST), 1 076 IU/L; alanine aminotransferase (ALT), 1 400 IU/L; total bilirubin (TB), 20.9 mg/dL; and prothrombin time rate (PT%), 46.9%. Acute hepatitis A (HA) was diagnosed based on negative hepatitis B surface antigen and hepatitis C virus RNA and positive immunoglobulin (Ig) M HA antibody, but elevation of anti-nuclear antigen (×320) and IgG (3 112 mg/dL) led to suspicion of autoimmune hepatitis (AIH). Plasma exchange was performed for 3 d from July 17, and steroid pulse therapy was performed for 3 d starting on July 18, followed by oral steroid therapy. Liver biopsy was performed on August 5, and the results confirmed acute hepatitis and mild chronic inflammation. Levels of AST and ALT normalized,so dose of oral steroid was markedly reduced. Steroid therapy was terminated after 4 mo, as the patient had glaucoma. Starting 3 mo after cessation of steroid therapy,levels of AST and ALT began to increase again. Another liver biopsy was performed and AIH was diagnosed based on serum data and biopsy specimen. Oral steroid therapy was reinitiated. Levels of AST and ALT again normalized.The present case was thus considered to represent AIH triggered by acute HA.

  2. Live recombinant BHV/BRSV vaccine

    NARCIS (Netherlands)

    Keil, G.M.; Rijsewijk, F.A.M.

    1998-01-01

    The present invention refers to synthetic Bovine Respiratory Syncytium virus genes. Also the invention relates to live attenuated Bovine Herpesvirus recombinants carrying such synthetic genes. Furthermore, the invention relates to vaccines based on these live attenuated recombinants, for the protect

  3. Recombinant innovation and endogenous technological transitions

    NARCIS (Netherlands)

    K. Frenken; L.R. Izquierdo; P. Zeppini

    2012-01-01

    We propose a model of technological transitions based on two different types of innovations. Branching innovations refer to technological improvements along a particular path, while recombinant innovations represent fusions of multiple paths. Recombinant innovations create "short-cuts" which reduce

  4. Enhanced mucosal immune responses induced by a combined candidate mucosal vaccine based on Hepatitis A virus and Hepatitis E virus structural proteins linked to tuftsin.

    Directory of Open Access Journals (Sweden)

    Yan Gao

    Full Text Available Hepatitis A virus (HAV and Hepatitis E virus (HEV are the most common causes of infectious hepatitis. These viruses are spread largely by the fecal-oral route and lead to clinically important disease in developing countries. To evaluate the potential of targeting hepatitis A and E infection simultaneously, a combined mucosal candidate vaccine was developed with the partial open reading frame 2 (ORF2 sequence (aa 368-607 of HEV (HE-ORF2 and partial virus protein 1 (VP1 sequence (aa 1-198 of HAV (HA-VP1, which included the viral neutralization epitopes. Tuftsin is an immunostimulatory peptide which can enhance the immunogenicity of a protein by targeting it to macrophages and dendritic cells. Here, we developed a novel combined protein vaccine by conjugating tuftsin to HE-ORF2 and HA-VP1 and used synthetic CpG oligodeoxynucleotides (ODNs as the adjuvant. Subsequent experiments in BALB/c mice demonstrated that tuftsin enhanced the serum-specific IgG and IgA antibodies against HEV and HAV at the intestinal, vaginal and pulmonary interface when delivered intranasally. Moreover, mice from the intranasally immunized tuftsin group (HE-ORF2-tuftsin + HA-VP1-tuftsin + CpG showed higher levels of IFN-γ-secreting splenocytes (Th1 response and ratio of CD4+/CD8+ T cells than those of the no-tuftsin group (HE-ORF2 + HA-VP1 + CpG. Thus, the tuftsin group generated stronger humoral and cellular immune responses compared with the no-tuftsin group. Moreover, enhanced responses to the combined protein vaccine were obtained by intranasal immunization compared with intramuscular injection. By integrating HE-ORF2, HA-VP1 and tuftsin in a vaccine, this study validated an important concept for further development of a combined mucosal vaccine against hepatitis A and E infection.

  5. Application of synchrotron-radiation-based x-ray microprobe techniques for the analysis of recombination activity of metals precipitated at Si/SiGe misfit dislocations

    Energy Technology Data Exchange (ETDEWEB)

    Vyvenko, O F [University of California, LBNL, 1 Cyclotron Rd, Berkeley, CA 94720 (United States); Buonassisi, T [University of California, LBNL, 1 Cyclotron Rd, Berkeley, CA 94720 (United States); Istratov, A A [University of California, LBNL, 1 Cyclotron Rd, Berkeley, CA 94720 (United States); Weber, E R [University of California, LBNL, 1 Cyclotron Rd, Berkeley, CA 94720 (United States); Kittler, M [IHP, Im Technologiepark 25, D-15236 Frankfurt (Oder) (Germany); Seifert, W [IHP, Im Technologiepark 25, D-15236 Frankfurt (Oder) (Germany)

    2002-12-09

    In this study we report application of synchrotron-radiation-based x-ray microprobe techniques (the x-ray-beam-induced current (XBIC) and x-ray fluorescence ({mu}-XRF) methods) to the analysis of the recombination activity and space distribution of copper and iron in the vicinity of dislocations in silicon/silicon-germanium structures. A combination of these two techniques enables one to study the chemical nature of the defects and impurities and their recombination activity in situ and to map metal clusters with a micron-scale resolution. XRF analysis revealed that copper formed clearly distinguishable precipitates along the misfit dislocations. A proportional dependence between the XBIC contrast and the number of copper atoms in the precipitates was established. In hydrogen-passivated iron-contaminated samples we observed clusters of iron precipitates which had no recombination activity detectable by the XBIC technique as well as iron clusters which were not completely passivated.

  6. Application of synchrotron-radiation-based x-ray microprobe techniques for the analysis of recombination activity of metals precipitated at Si/SiGe misfit dislocations

    CERN Document Server

    Vyvenko, O F; Istratov, A A; Weber, E R; Kittler, M; Seifert, W

    2002-01-01

    In this study we report application of synchrotron-radiation-based x-ray microprobe techniques (the x-ray-beam-induced current (XBIC) and x-ray fluorescence (mu-XRF) methods) to the analysis of the recombination activity and space distribution of copper and iron in the vicinity of dislocations in silicon/silicon-germanium structures. A combination of these two techniques enables one to study the chemical nature of the defects and impurities and their recombination activity in situ and to map metal clusters with a micron-scale resolution. XRF analysis revealed that copper formed clearly distinguishable precipitates along the misfit dislocations. A proportional dependence between the XBIC contrast and the number of copper atoms in the precipitates was established. In hydrogen-passivated iron-contaminated samples we observed clusters of iron precipitates which had no recombination activity detectable by the XBIC technique as well as iron clusters which were not completely passivated.

  7. Predictors of response to pegylated interferon in chronic hepatitis B: a real-world hospital-based analysis

    Science.gov (United States)

    Wang, Yin-Chen; Yang, Sien-Sing; Su, Chien-Wei; Wang, Yuan-Jen; Lee, Kuei-Chuan; Huo, Teh-Ia; Lin, Han-Chieh; Huang, Yi-Hsiang

    2016-01-01

    Information on the efficacy of pegylated interferon (PEG-IFN) treatment of chronic hepatitis B (CHB) patients and predictors of the response based on real-world data is limited. Consecutive 201 patients who underwent PEG-IFN treatment for CHB were reviewed. A virological response (VR) was defined as a serum HBV DNA of <2000 IU/mL, and a combined response (CR) was defined a VR accompanied by serological response for hepatitis B e antigen (HBeAg)-positive CHB. For HBeAg-positive CHB patients, the HBeAg seroconversion rate and CR rate were 30.5% and 21.2% at 48 weeks after end of treatment (EOT), respectively. Baseline alanine aminotransferase (ALT) level was associated with HBeAg seroconversion, while baseline hepatitis B s antigen (HBsAg) levels of <250 IU/mL and HBV DNA <2.5 × 107 IU/mL were strongly associated with sustained off-treatment CR. For HBeAg-negative CHB, the VR rates were 85.5%, and 27.7% at EOT, and 48 weeks after EOT, respectively; a baseline HBsAg <1,250 IU/mL was associated with sustained off-treatment VR. PEG-IFN treatment has durable HBeAg seroconversion in HBeAg-positive CHB, but results in a high risk of relapse among HBeAg-negative CHB patients. Pre-treatment HBsAg level is an important predictor of VR in CHB patients undergoing PEG-IFN treatment. PMID:27405043

  8. Recombination in ionized gases

    International Nuclear Information System (INIS)

    In this paper it is shown how capture-stabilized methodology (both macroscopic and microscopic) can provide a generic basis for a unified treatment of all of the above recombination mechanisms. A new semiclassical theory of dissociative recombination is also presented in an effort to gain further insight into the physics not included in the first-order treatment and difficult to extract from numerical quantal treatments based on configuration mixing and on multichannel quantum defect theory. A simple analytical expression more accurate than the standard first-order result is obtained for the cross section σ and rate coefficient α. (author)

  9. Recombineering Pseudomonas syringae

    Science.gov (United States)

    Here we report the identification of functions that promote genomic recombination of linear DNA introduced into Pseudomonas cells by electroporation. The genes encoding these functions were identified in Pseudomonas syringae pv. syringae B728a based on similarity to the lambda Red Exo/Beta and RecE...

  10. The epidemiology of viral hepatitis in Qatar

    Directory of Open Access Journals (Sweden)

    Bener Abdulbari

    2009-01-01

    Full Text Available Viral hepatitis is a major public health problem in many countries all over the world and especially in Middle East, Asia, East-Europe, and Africa. The aim of our study was to assess the incidence of viral hepatitis A, B and C in Qatar and compare it with other countries. This is a retrospective cohort study, which was conducted at Hamad General Hospital, State of Qatar from 2002-2006. Patients who were screened and diagnosed with viral hepatitis were included in this study. The diagnostic classification of definite viral hepatitis was made in accordance with criteria based on the International Classification of Disease tenth revision (ICD-10. A total of 527 cases of hepatitis C, 396 cases of hepatitis B, 162 cases of hepatitis A and 108 cases of unspecified were reported during the year 2006. Reported incidence rate per 10,000 populations during the year 2006 for hepatitis A was 1.9, hepatitis B 4.7, and Hepatitis C 6.3. The proportion of hepatitis B and C was significantly higher in male population than females across the years (2002-2006. Hepatitis A was more prevalent in children below 15 years (72.3%, hepatitis B in adults aged above 15 years, and hepatitis C in the population above 35 years of age. The incidence of hepatitis A has been declining in Qataris and increasing in expatriates. There was a significant relationship in gender and age group of the patients with hepatitis A, B and C. We conclude that hepatitis has become a national health issue in Qatar. The incidence rate of hepatitis in Qatar is comparable to its neighboring countries, United Arab Emirates and Saudi Arabia. There is a need for further research on hepatitis and the associated risk factors.

  11. Forecasting model for the incidence of hepatitis A based on artificial neural network

    Institute of Scientific and Technical Information of China (English)

    Peng Guan; De-Sheng Huang; Bao-Sen Zhou

    2004-01-01

    AIM: To study the application of artificial neural network (ANN) in forecasting the incidence of hepatitis A, which had an autoregression phenomenon.METHODS: The data of the incidence of hepatitis A in Liaoning Province from 1981 to 2001 were obtained from Liaoning Disease Control and Prevention Center. We used the autoregressive integrated moving average (ARIMA)model of time series analysis to determine whether there was any autoregression phenomenon in the data. Then the data of the incidence were switched into [0,1] intervals as the network theoretical output. The data from 1981 to 1997 were used as the training and verifying sets and the data from 1998 to 2001 were made up into the test set.STATISTICA neural network (ST NN) was used to construct,train and simulate the artificial neural network.RESULTS: Twenty-four networks were tested and seven were retained. The best network we found had excellent performance, its regression ratio was 0.73, and its correlatior, was 0.69. There were 2 input variables in the network, one was AR(1), and the other was time. The number of units in hidden layer was 3. In ARIMA time series analysis results, the best model was first order autoregression without difference and smoothness. The total sum square error of the ANN model was 9 090.21, the sum square error of the training set and testing set was 8 377.52 and 712.69,respectively, they were all less than that of ARIMA model.The corresponding value of ARIMA was 12 291.79, 8 944.95and 3 346.84, respectively. The correlation coefficient of nonlinear regression (RNL) of ANN was 0.71, while the RNL of ARIMA linear autoregression model was 0.66.CONCLUSION: ANN is superior to conventional methods in forecasting the incidence of hepatitis A which has an autoregression phenomenon.

  12. Current progress in the development of therapeutic vaccines for chronic hepatitis B virus infection

    Directory of Open Access Journals (Sweden)

    Faezeh Ghasemi

    2016-07-01

    Full Text Available Chronic hepatitis B is still a major public health issue despite the successful prophylactic vaccination attempts. Chronicity of hepatitis B virus(HBV is mainly due to its ability to debilitate host's immune system. Therefore, major measures have been taken to stop this process and help patients with chronic hepatitis B infection recover from their illness. While satisfactory results have been achieved using preventive HBV vaccines, a reliable and effective therapeutic treatment is still in need of extensive studies. Current treatments for chronic hepatitis B include direct antiviral agents and nucleoside/nucleotide analogs, which are not always effective and are also costly. In addition, due to the fact that chronic HBV is responsible for debilitation of the immune system, studies have focused on developing therapeutic vaccines to help host's immune system recover and limit the infection. Several approaches including but not restricted to recombinant peptide-based, DNA-based, viral vector-based, and cell-based approaches are currently in use to develop therapeutic vaccines against the chronic form of HBV infection. In the current review, the authors will first discuss the role of the immune system in chronic hepatitis B infection and will then focus on latest advancements in therapeutic vaccination of HBV especially the clinical trials that have been carried out so far.

  13. Current progress in the development of therapeutic vaccines for chronic hepatitis B virus infection.

    Science.gov (United States)

    Ghasemi, Faezeh; Rostami, Sina; Ghayour-Mobarhan, Majid; Meshkat, Zahra

    2016-07-01

    Chronic hepatitis B is still a major public health issue despite the successful prophylactic vaccination attempts. Chronicity of hepatitis B virus (HBV) is mainly due to its ability to debilitate host's immune system. Therefore, major measures have been taken to stop this process and help patients with chronic hepatitis B infection recover from their illness. While satisfactory results have been achieved using preventive HBV vaccines, a reliable and effective therapeutic treatment is still in need of extensive studies. Current treatments for chronic hepatitis B include direct antiviral agents and nucleoside/nucleotide analogs, which are not always effective and are also costly. In addition, due to the fact that chronic HBV is responsible for debilitation of the immune system, studies have focused on developing therapeutic vaccines to help host's immune system recover and limit the infection. Several approaches including but not restricted to recombinant peptide-based, DNA-based, viral vector-based, and cell-based approaches are currently in use to develop therapeutic vaccines against the chronic form of HBV infection. In the current review, the authors will first discuss the role of the immune system in chronic hepatitis B infection and will then focus on latest advancements in therapeutic vaccination of HBV especially the clinical trials that have been carried out so far. PMID:27635192

  14. Targeting c-kit receptor in neuroblastomas and colorectal cancers using stem cell factor (SCF)-based recombinant bacterial toxins.

    Science.gov (United States)

    Choudhary, Swati; Pardo, Alessa; Rosinke, Reinhard; Batra, Janendra K; Barth, Stefan; Verma, Rama S

    2016-01-01

    Autocrine activation of c-kit (KIT receptor tyrosine kinase) has been postulated to be a potent oncogenic driver in small cell lung cancer, neuroblastoma (NB), and poorly differentiated colorectal carcinoma (CRC). Although targeted therapy involving tyrosine kinase inhibitors (TKIs) such as imatinib mesylate is highly effective for gastrointestinal stromal tumor carrying V560G c-kit mutation, it does not show much potential for targeting wild-type KIT (WT-KIT). Our study demonstrates the role of stem cell factor (SCF)-based toxin conjugates for targeting WT-KIT-overexpressing malignancies such as NBs and CRCs. We constructed SCF-based recombinant bacterial toxins by genetically fusing mutated form of natural ligand SCF to receptor binding deficient forms of Diphtheria toxin (DT) or Pseudomonas exotoxin A (ETA') and evaluated their efficacy in vitro. Efficient targeting was achieved in all receptor-positive neuroblastoma (IMR-32 and SHSY5Y) and colon cancer cell lines (COLO 320DM, HCT 116, and DLD-1) but not in receptor-negative breast carcinoma cell line (MCF-7) thereby proving specificity. While dose- and time-dependent cytotoxicity was observed in both neuroblastoma cell lines, COLO 320DM and HCT 116 cells, only an anti-proliferative effect was observed in DLD-1 cells. We prove that these novel targeting agents have promising potential as KIT receptor tyrosine kinase targeting system.

  15. Pharmacogenetics of hepatitis C: transition from interferon-based therapies to direct-acting antiviral agents

    Directory of Open Access Journals (Sweden)

    Kamal SM

    2014-06-01

    Full Text Available Sanaa M Kamal1,21Department of Medicine, Division of Hepatology, Gastroenterology and Tropical Medicine, Ain Shams Faculty of Medicine, Cairo, Egypt, 2Department of Medicine, Salman Bin Abdul Aziz College of Medicine, Kingdom of Saudi ArabiaAbstract: Hepatitis C virus (HCV has emerged as a major viral pandemic over the past two decades, infecting 170 million individuals, which equates to approximately 3% of the world's population. The prevalence of HCV varies according to geographic region, being highest in developing countries such as Egypt. HCV has a high tendency to induce chronic progressive liver damage in the form of hepatic fibrosis, cirrhosis, or liver cancer. To date, there is no vaccine against HCV infection. Combination therapy comprising PEGylated interferon-alpha and ribavirin has been the standard of care for patients with chronic hepatitis C for more than a decade. However, many patients still do not respond to therapy or develop adverse events. Recently, direct antiviral agents such as protease inhibitors, polymerase inhibitors, or NS5A inhibitors have been used to augment PEGylated interferon and ribavirin, resulting in better efficacy, better tolerance, and a shorter treatment duration. However, most clinical trials have focused on assessing the efficacy and safety of direct antiviral agents in patients with genotype 1, and the response of other HCV genotypes has not been elucidated. Moreover, the prohibitive costs of such triple therapies will limit their use in patients in developing countries where most of the HCV infection exists. Understanding the host and viral factors associated with viral clearance is necessary for individualizing therapy to maximize sustained virologic response rates, prevent progression to liver disease, and increase the overall benefits of therapy with respect to its costs. Genome wide studies have shown significant associations between a set of polymorphisms in the region of the interleukin-28B (IL

  16. Regulation of Meiotic Recombination

    Energy Technology Data Exchange (ETDEWEB)

    Gregory p. Copenhaver

    2011-11-09

    for assaying recombination using tetrad analysis in a higher eukaryotic system (6). This system enabled the measurement of the frequency and distribution of recombination events at a genome wide level in wild type Arabidopsis (7), construction of genetic linkage maps which include positions for each centromere (8), and modeling of the strength and pattern of interference (9). This proposal extends the use of tetrad analysis in Arabidopsis by using it as the basis for assessing the phenotypes of mutants in genes important for recombination and the regulation of crossover interference and performing a novel genetic screen. In addition to broadening our knowledge of a classic genetic problem - the regulation of recombination by crossover interference - this proposal also provides broader impact by: generating pedagogical tools for use in hands-on classroom experience with genetics, building interdisciplinary collegial partnerships, and creating a platform for participation by junior scientists from underrepresented groups. There are three specific aims: (1) Isolate mutants in Arabidopsis MUS81 homologs using T-DNA and TILLING (2) Characterize recombination levels and interference in mus81 mutants (3) Execute a novel genetic screen, based on tetrad analysis, for genes that regulate meiotic recombination

  17. Comparison of various methods for breakage of recombinant Hansenula polymorpha cells for expression of hepatitis B surface antigen%表达乙型肝炎表面抗原重组汉逊酵母细胞破碎方法的比较

    Institute of Scientific and Technical Information of China (English)

    王曦; 李津; 李彩梅; 劳文燕; 许宁; 刘英微; 张德有; 马锐; 杨旭琴; 朱亭玉

    2012-01-01

    Objective To compare the breakage effects of recombinant Hansenula polymorpha (HP) cells for expression of hepatitis B surface antigen (HBsAg) by bead-milling,low-temperature high-pressure homogenization and Yeast Buster cell lysate treatment. Methods Recombinant HP cells for expression of HBsAg were broken by bead-milling,low-temperature high-pressure homogenization and Yeast Buster cell lysate treatment respectively,using recombinant Saccharomyces cerevisiae cells as control. The breakage rates of cells and the releases of protein and HBsAg were determined,based on which the effects of three methods were compared. Results The components in either jars or beads were abraded into the cells broken by bead-milling. Agate jars and zirconia beads produced fewer impurities,by which the breakage rate of HP cells was only about 60%. The highest protein content of HP cells broken by bead-milling with agate jars and glass beads was 2. 71 mg/ml,while the highest release of HBsAg was 180. 04 μg/ml. The breakage rate of recombinant HP cells by low-temperature high-pressure homogenization was 71. 18% at most,while the highest protein content was 6. 72 mg/ml,and the highest release of HBsAg was 350. 63 μg/ml. However,the breakage rate of HP cells by Yeast Buster cell lysate was 60. 30%,while significant differences were observed in protein contents and HBsAg releases in the cells broken for various times. Conclusion Compared with bead-milling and Yeast Buster cell lysate treatment,low-temperature high pressure homogenization is more suitable for breakage of recombinant HP cells for expression of HBsAg.%目的 比较球磨法、低温超高压破碎法和Yeast Buster酵母细胞裂解液法破碎表达乙型肝炎表面抗原(HBsAg)的重组汉逊酵母(Hamenula polymorpha,HP)细胞的效果.方法 分别采用球磨法、低温超高压破碎法和细胞裂解液法破碎表达HBsAg的重组HP细胞和重组酿酒酵母(Saccharomyces cerevisiea,SC)细胞,测定其细胞破碎

  18. MODEST: a web-based design tool for oligonucleotide-mediated genome engineering and recombineering

    DEFF Research Database (Denmark)

    Bonde, Mads; Klausen, Michael S.; Anderson, Mads Valdemar;

    2014-01-01

    , which confers the corresponding genetic change, is performed manually. To address these challenges, we have developed the MAGE Oligo Design Tool (MODEST). This web-based tool allows designing of MAGE oligos for (i) tuning translation rates by modifying the ribosomal binding site, (ii) generating...... as combinatorial cell libraries. Manual design of oligonucleotides for these approaches can be tedious, time-consuming, and may not be practical for larger projects targeting many genomic sites. At present, the change from a desired phenotype (e.g. altered expression of a specific protein) to a designed MAGE oligo...... translational gene knockouts and (iii) introducing other coding or non-coding mutations, including amino acid substitutions, insertions, deletions and point mutations. The tool automatically designs oligos based on desired genotypic or phenotypic changes defined by the user, which can be used for high...

  19. A recombination based method to rapidly assess specificity of two-hybrid clones in yeast.

    OpenAIRE

    Petermann, R; Mossier, B M; Aryee, D N; Kovar, H.

    1998-01-01

    The yeast two-hybrid system is frequently used to identify protein-protein interactions. Confirming the specificity of candidate clones requires separation and isolation of yeast plasmids, propagation in bacteria and testing combinations of DNA-binding and activation domain hybrids in yeast. In order to simplify this procedure, we developed a rapid method based on PCR amplification of library insert DNAs and in vivo cloning into the activation domain hybrid vector. Reporter gene activity is a...

  20. Immunogenecity of hepatitis B vaccine in Mazandaran medical sciences students-2004

    OpenAIRE

    Ajami, A.; F. Abediyan

    2006-01-01

    Background and purpose: Vaccination is the most effeetive method of prophylaxy in high risk group of hepatitis B. Some individuals faile to respond to triple doses of vaccine which is potentially dangerous. Therefore, immunogenecity of triple doses recombinant hepatitis B vaccine was evaluated in a high risk group (medical sciences students).Materials and Methods: 193 healthy medical sciences students immunized with triple doses of recombinant hepatitis B vaccine (0,1,6 months). 1-2 months af...

  1. Clinical Effectiveness and Cost Effectiveness of Tailoring Chronic Hepatitis C Treatment with Peginterferon Alpha-2b Plus Ribavirin to HCV Genotype and Early Viral Response: A Decision Analysis Based on German Guidelines

    OpenAIRE

    Uwe Siebert; Gaby Sroczynski; Pamela Aidelsburger; Siegbert Rossol; Jurgen Wasem; Manns, Michael P.; MCHUTCHISON, JOHN G.; Wong, John B.

    2009-01-01

    Background Recently developed German guidelines for antiviral treatment in patients with chronic hepatitis C recommend basing drug dosage, intended treatment duration and early stopping rules on the genotype of the hepatitis C virus and early viral responses to treatment. Abstract: Objectives To evaluate effectiveness and cost effectiveness of different antiviral treatment strategies including the German guidelines, for chronic hepatitis C. Abstract: Methods A validated lifetime Markov model ...

  2. Poor uptake of hepatitis B immunization amongst hospital-based health care staff.

    Science.gov (United States)

    Burden, A D; Whorwell, P J

    1991-03-01

    The uptake of hepatitis B vaccine was assessed amongst 100 medical and 100 nursing staff in a teaching hospital with a policy of recommending to those at risk that they should seek immunization from their general practitioners. Sixteen per cent of nurses and 31% of doctors had completed a course of immunization with confirmation of seroconversion. An additional 9% and 18% respectively had been immunized without post-immunization serology. Ninety three per cent of nurses and 61% of doctors who had not been immunized would like to receive the vaccine. The commonest reasons for non-immunization amongst nurses were fear of vaccine and lack of advice, and amongst doctors, apathy and difficulty in obtaining the vaccine. Eighty seven per cent of medical staff and 57% of nurses had a history of needle stick injury. The low rates of vaccine uptake in this study combined with the high incidence of needle stick injury calls for a reappraisal of present hepatitis B vaccination programmes in hospitals. In particular the abrogation of responsibility for immunization to general practitioners is probably a major disincentive to potential vaccines.

  3. Serology based disease status of Pakistani population infected with Hepatitis B virus

    Directory of Open Access Journals (Sweden)

    Sharif Salmaan

    2007-06-01

    Full Text Available Abstract Background The infection rate of hepatitis B virus is continuously increasing in Pakistan. Therefore, a comprehensive study of epidemiological data is the need of time. Methods A total of 1300 individuals were screened for HBV infection markers including HBsAg, anti-HBsAg, HBeAg and anti-HBcAg. The association of these disease indicators was compared with patients' epidemiological characteristics like age, socio-economic status and residential area to analyze and find out the possible correlation among these variables and the patients disease status. Results 52 (4% individuals were found positive for HBsAg with mean age 23.5 ± 3.7 years. 9.30%, 33.47% and 12% individuals had HBeAg, antibodies for HBsAg, and antibodies for HBcAg respectively. HBsAg seropositivity rate was significantly associated (p = 0.03 with the residing locality indicating high infection in rural areas. Antibodies titer against HBsAg decreased with the increasing age reflecting an inverse correlation. Conclusion Our results indicate high prevalence rate of Hepatitis B virus infection and nationwide vaccination campaigns along with public awareness and educational programs are needed to be practiced urgently.

  4. Therapeutic effects of recombinant hepatitis B virus vaccine combined with interferon α-1 b in patients with chronic hepatitis B%重组酵母乙型肝炎疫苗联合干扰素α-1 b治疗慢性乙型肝炎的疗效

    Institute of Scientific and Technical Information of China (English)

    徐启桓; 陈菊梅; 张晓红; 谢冬英; 李建国; 崇雨田; 杨林; 陆玮伦; 高志良; 田德英

    2008-01-01

    Objective To investigate the therapeutic effects of recombinant yeast hepatitis B virus(HBV)vaccine combined with interferon(IFN)α-1b and determine the rational dosage of HBV vaccine for the further clinical study with larger sample.Methods Two hundreds and sixteen patients with chronic hepatitis B(CHB)were enrolled in this randomized,multi-center,double-blinded and placebo-controlled clinical trial.All the subjects were not treated with antiviral drugs within 6 months and randomly divided into 90μg,60μg and placebo groups with a ratio of 1:1:1.All the patients were intramuscularly administrated with 90μg or 60μg recombinant HBV vaccine or placebo at week 0,2,6,10,14,18,22,respectively.Meanwhile,they were also treated with IFNα-1b 50μg,3 times a wcek for 24 weeks.All patients were followed up for 24 weeks after withdrawal of anti-HBV therapy.Serum HBV DNA level,HBeAg titer and liver function were monitored frequently.Interferon-γ secreting lymphoeytes were detected by Enzyme-linked immunospot(ELISPOT)in part of the patients.Results The serum HBV DNA levels were(6.03±1.79),(5.52±1.82)and(6.29±1.70)log10 copy/mL at week 24 of treatment in high dose,low dose and placebo groups,respectively (P=0.458).And the serum HBV DNA levels were(5.92±1.98),(5.49±1.99)and(6.16±1.76)log10 copy/mL at weck 24 after withdrawal of treatment,respectively(P=0.720).The rates of patients whose HBV DNA<1×105 copy/mL in these three groups were 30.4%,39.4% and 20.8% at week 24 of treatment,respectively and there was significant difference between high dose group and low dose group(P=0.015).The rate of patients whose HBV DNA<1×105 copy/mL at week 24 after withdrawal was highest in low dose group,but no significant differences before and after treatment in these three groups(P=0.257).The HBV DNA negative rates were 17.4%,25.4% and 6.9% in these three groups,respectively,which were significantly different(P=0.012).At week 24 of treatment and week 24 after withdrawal of

  5. Hybridization-based antibody cDNA recovery for the production of recombinant antibodies identified by repertoire sequencing

    OpenAIRE

    Valdés-Alemán, Javier; Téllez-Sosa, Juan; Ovilla-Muñoz, Marbella; Godoy-Lozano, Elizabeth; Velázquez-Ramírez, Daniel; Valdovinos-Torres, Humberto; Gómez-Barreto, Rosa E; Martinez-Barnetche, Jesús

    2013-01-01

    High-throughput sequencing of the antibody repertoire is enabling a thorough analysis of B cell diversity and clonal selection, which may improve the novel antibody discovery process. Theoretically, an adequate bioinformatic analysis could allow identification of candidate antigen-specific antibodies, requiring their recombinant production for experimental validation of their specificity. Gene synthesis is commonly used for the generation of recombinant antibodies identified in silico. Novel ...

  6. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: Lessons from the past and novel mechanisms of action for the future.

    Science.gov (United States)

    Valenta, Rudolf; Campana, Raffaela; Focke-Tejkl, Margit; Niederberger, Verena

    2016-02-01

    In the past, the development of more effective, safe, convenient, broadly applicable, and easy to manufacture vaccines for allergen-specific immunotherapy (AIT) has been limited by the poor quality of natural allergen extracts. Progress made in the field of molecular allergen characterization has now made it possible to produce defined vaccines for AIT and eventually for preventive allergy vaccination based on recombinant DNA technology and synthetic peptide chemistry. Here we review the characteristics of recombinant and synthetic allergy vaccines that have reached clinical evaluation and discuss how molecular vaccine approaches can make AIT more safe and effective and thus more convenient. Furthermore, we discuss how new technologies can facilitate the reproducible manufacturing of vaccines of pharmaceutical grade for inhalant, food, and venom allergens. Allergy vaccines in clinical trials based on recombinant allergens, recombinant allergen derivatives, and synthetic peptides allow us to target selectively different immune mechanisms, and certain of those show features that might make them applicable not only for therapeutic but also for prophylactic vaccination.

  7. Construction and expression of a recombinant eukaryotic expression plasmid containing the preS1-preS2-S genes of hepatitis B virus and the granulocyte-macrophage colony stimulating factor gene: A study of its immunomodulatory effects.

    Science.gov (United States)

    Gong, Jun-Yuan; Liu, Xin; Dong, Yan; Zhou, Tian-Hong; Li, Jun-Wu

    2013-03-01

    A total of 10-20% of the population remains unresponsive or weakly responsive to hepatitis B vaccine, which is composed of hepatitis B surface antigen HBsAg (S protein). Therefore, it is necessary to develop a hepatitis B vaccine with a better penetrating and responsive rate. In the present study, a plasmid pVAX1-L-GM was constructed and its immunomodulatory effect of as hepatitis B virus (HBV) DNA vaccine was analyzed through the immunization of BALB/c mice. Immune responses were measured after immunization by anti-HBsAg, proliferation of splenocytes, the number of CD4(+) and CD8(+) molecules, CTL cytotoxicity, cytokines of IFN-γ and IL-2 secretion assays. Following the immunization, mice in the pVAX1-L-GM group produced antibody 2 weeks earlier compared to the control plasmid pVAX1 and pVAX1HBsAg groups and antibody levels showed significant differences. Enhanced HBsAg-specific splenocyte proliferation as well as specific cytotoxic activities of splenic CTLs were also detected. Furthermore, pVAX1-L-GM plasmid increased the number of CD4(+) and CD8(+) molecules on the surface of the spleen T cell and the level of IFN-γ, IL-2 secretion. pVAX1-L-GM induced a specific immune response in mice and enhanced the immune effect. Thus, a foundation was laid for developing immunogenicity of a better prevention and treatment of HBV via a hepatitis B vaccine. PMID:24648930

  8. A Puzzle-Based Genetic Algorithm with Block Mining and Recombination Heuristic for the Traveling Salesman Problem

    Institute of Scientific and Technical Information of China (English)

    Pei-Chann Chang; Wei-Hsiu Huang; Zhen-Zhen Zhang

    2012-01-01

    In this research,we introduce a new heuristic approach using the concept of ant colony optimization (ACO)to extract patterns from the chromosomes generated by previous generations for solving the generalized traveling salesman problem.The proposed heuristic is composed of two phases.In the first phase the ACO technique is adopted to establish an archive consisting of a set of non-overlapping blocks and of a set of remaining cities (nodes) to be visited.The second phase is a block recombination phase where the set of blocks and the rest of cities are combined to form an artificial chromosome.The generated artificial chromosomes (ACs) will then be injected into a standard genetic algorithm (SGA) to speed up the convergence.The proposed method is called "Puzzle-Based Genetic Algorithm" or "p-ACGA".We demonstrate that p-ACGA performs very well on all TSPLIB problems,which have been solved to optimality by other researchers.The proposed approach can prevent the early convergence of the genetic algorithm (GA) and lead the algorithm to explore and exploit the search space by taking advantage of the artificial chromosomes.

  9. Immunity conferred by an experimental vaccine based on the recombinant PCV2 Cap protein expressed in Trichoplusia ni-larvae.

    Science.gov (United States)

    Pérez-Martín, Eva; Gómez-Sebastián, Silvia; Argilaguet, Jordi M; Sibila, Marina; Fort, María; Nofrarías, Miquel; Kurtz, Sherry; Escribano, José M; Segalés, Joaquim; Rodríguez, Fernando

    2010-03-01

    Porcine circovirus type 2 (PCV2) vaccination has been recently included as a measure to control postweaning multisystemic wasting syndrome (PMWS) in the field. Aiming to obtain a more affordable vaccine to be extensively implemented in the field, a highly efficient non-fermentative expression platform based on Trichoplusia ni (T. ni) larvae was used to produce a baculovirus-derived recombinant PCV2 Cap protein (rCap) for vaccine purposes. Vaccination of pigs with rCap induced solid protection against PCV2 experimental infection, inhibiting both the viremia and the viral shedding very efficiently. The protection afforded by the rCap vaccine strongly correlated with the induction of specific humoral immune responses, even in the presence of PCV2-specific maternal immunity, although cellular responses also seemed to play a partial role. In summary, we have shown that rCap expressed in T. ni larvae could be a cost-effective PCV2 vaccine candidate to be tested under field conditions. PMID:20056179

  10. Yeast homologous recombination-based promoter engineering for the activation of silent natural product biosynthetic gene clusters.

    Science.gov (United States)

    Montiel, Daniel; Kang, Hahk-Soo; Chang, Fang-Yuan; Charlop-Powers, Zachary; Brady, Sean F

    2015-07-21

    Large-scale sequencing of prokaryotic (meta)genomic DNA suggests that most bacterial natural product gene clusters are not expressed under common laboratory culture conditions. Silent gene clusters represent a promising resource for natural product discovery and the development of a new generation of therapeutics. Unfortunately, the characterization of molecules encoded by these clusters is hampered owing to our inability to express these gene clusters in the laboratory. To address this bottleneck, we have developed a promoter-engineering platform to transcriptionally activate silent gene clusters in a model heterologous host. Our approach uses yeast homologous recombination, an auxotrophy complementation-based yeast selection system and sequence orthogonal promoter cassettes to exchange all native promoters in silent gene clusters with constitutively active promoters. As part of this platform, we constructed and validated a set of bidirectional promoter cassettes consisting of orthogonal promoter sequences, Streptomyces ribosome binding sites, and yeast selectable marker genes. Using these tools we demonstrate the ability to simultaneously insert multiple promoter cassettes into a gene cluster, thereby expediting the reengineering process. We apply this method to model active and silent gene clusters (rebeccamycin and tetarimycin) and to the silent, cryptic pseudogene-containing, environmental DNA-derived Lzr gene cluster. Complete promoter refactoring and targeted gene exchange in this "dead" cluster led to the discovery of potent indolotryptoline antiproliferative agents, lazarimides A and B. This potentially scalable and cost-effective promoter reengineering platform should streamline the discovery of natural products from silent natural product biosynthetic gene clusters.

  11. A recombinant nucleocapsid protein-based indirect enzyme-linked immunosorbent assay to detect antibodies against porcine deltacoronavirus.

    Science.gov (United States)

    Su, Mingjun; Li, Chunqiu; Guo, Donghua; Wei, Shan; Wang, Xinyu; Geng, Yufei; Yao, Shuang; Gao, Jing; Wang, Enyu; Zhao, Xiwen; Wang, Zhihui; Wang, Jianfa; Wu, Rui; Feng, Li; Sun, Dongbo

    2016-05-01

    Recently, porcine deltacoronavirus (PDCoV) has been proven to be associated with enteric disease in piglets. Diagnostic tools for serological surveys of PDCoV remain in the developmental stage when compared with those for other porcine coronaviruses. In our study, an indirect enzyme-linked immunosorbent assay (ELISA) (rPDCoV-N-ELISA) was developed to detect antibodies against PDCoV using a histidine-tagged recombinant nucleocapsid (N) protein as an antigen. The rPDCoV-N-ELISA did not cross-react with antisera against porcine epidemic diarrhea virus, swine transmissible gastroenteritis virus, porcine group A rotavirus, classical swine fever virus, porcine circovirus-2, porcine pseudorabies virus, and porcine reproductive and respiratory syndrome virus; the receiver operating characteristic (ROC) curve analysis revealed 100% sensitivity and 90.4% specificity of the rPDCoV-N-ELISA based on samples of known status (n=62). Analyses of field samples (n=319) using the rPDCoV-N-ELISA indicated that 11.59% of samples were positive for antibodies against PDCoV. These data demonstrated that the rPDCoV-N-ELISA can be used for epidemiological investigations of PDCoV and that PDCoV had a low serum prevalence in pig population in Heilongjiang province, northeast China. PMID:26668175

  12. DEVELOPMENT OF RECOMBINANT VACCINE AGAINST A(H1N1) 2009 INFLUENZA BASED ON VIRUS-LIKE NANOPARTICLES CARRYING THE EXTRACELLULAR DOMAIN OF M2 PROTEIN

    OpenAIRE

    Kotlyarov, R.; Kuprianov, V.; Migunov, A.; Stepanova, L.; Tsybalova, L.; Kiselev, O.; Ravin, N.; Skryabin, K.

    2010-01-01

    The conventional vaccines currently being used to deal with influenza are based on a virus obtained in chicken embryos or its components. The high variability of the major immunogenic surface proteins – hemagglutinin and neuraminidase–require the development of strain–specific vaccines that match the antigenic specificity of a newly emerging virus. Recombinant vaccines based on single viral proteins that could be easily produced in standard expression systems are attractive alternatives to tr...

  13. Order of amino acids in C-terminal cysteine-containing peptide-based chelators influences cellular processing and biodistribution of 99mTc-labeled recombinant Affibody molecules.

    Science.gov (United States)

    Altai, Mohamed; Wållberg, Helena; Orlova, Anna; Rosestedt, Maria; Hosseinimehr, Seyed Jalal; Tolmachev, Vladimir; Ståhl, Stefan

    2012-05-01

    Affibody molecules constitute a novel class of molecular display selected affinity proteins based on non-immunoglobulin scaffold. Preclinical investigations and pilot clinical data have demonstrated that Affibody molecules provide high contrast imaging of tumor-associated molecular targets shortly after injection. The use of cysteine-containing peptide-based chelators at the C-terminus of recombinant Affibody molecules enabled site-specific labeling with the radionuclide 99mTc. Earlier studies have demonstrated that position, composition and the order of amino acids in peptide-based chelators influence labeling stability, cellular processing and biodistribution of Affibody molecules. To investigate the influence of the amino acid order, a series of anti-HER2 Affibody molecules, containing GSGC, GEGC and GKGC chelators have been prepared and characterized. The affinity to HER2, cellular processing of 99mTc-labeled Affibody molecules and their biodistribution were investigated. These properties were compared with that of the previously studied 99mTc-labeled Affibody molecules containing GGSC, GGEC and GGKC chelators. All variants displayed picomolar affinities to HER2. The substitution of a single amino acid in the chelator had an appreciable influence on the cellular processing of 99mTc. The biodistribution of all 99mTc-labeled Affibody molecules was in general comparable, with the main difference in uptake and retention of radioactivity in excretory organs. The hepatic accumulation of radioactivity was higher for the lysine-containing chelators and the renal retention of 99mTc was significantly affected by the amino acid composition of chelators. The order of amino acids influenced renal uptake of some conjugates at 1 h after injection, but the difference decreased at later time points. Such information can be helpful for the development of other scaffold protein-based imaging and therapeutic radiolabeled conjugates.

  14. Delta Hepatitis

    OpenAIRE

    Fulya Gunsar

    2012-01-01

    Hepatitis delta virus (HDV) is a defective RNA virus that requires HBsAg for replication and transmission. It can cause acute or chronic hepatitis. Chronic infection with HDV is one of the most severe and difficult to treat forms of viral hepatitis. It has been estimated that there is a total of 15-20 million HDV carriers in the world. This review focuses on two fundamental aspects of HDV infection. On the one hand, epidemiological data are summarized, which are essential to understand the re...

  15. 60 μg重组乙型肝炎疫苗(酿酒酵母)对无应答人群免疫效果观察%Observation on Immunological Effect of 60μg Recombinant Hepatitis B Vaccine (Saccharomyces cerevisiae) on the Non-response Crowd

    Institute of Scientific and Technical Information of China (English)

    肖业荣; 李强

    2013-01-01

    Objective To explore the immune success rate and effect of 60μg recombinant hepatitis B vaccine ( Saccharo-myces cereuisiae) on the HBV vaccine non- response crowd. Methods Forty volunteers aged above 17 years and vaccinated three hepatitis B vaccines for normal dose and procedure but without hepatitis B surface antibody were selected in Taojiang County. The serum samples of the volunteers were collected one month after inoculating one dose of 60μg reeombinant hepatitis B vaccine which produced by Shenzhen Kangtai Biological Products Co. Ltd. (SKTB), and then serum immunological observation was conducted. Results No local reaction and systemic reaction occurred in all volunteers. One month after inoculating one dose of 60μg recombinant hepatitis B vaccine, the positive rate of anti - HBs was 97. 5% and the geometric mean titer for anti - HBs was 50.657 mlU/ml. Conclusions The results suggest that 60μg reeombinant hepatitis B vaccine (Saccharomyces cerevisiae) has a good safety profile and exerts success in protection. The vaccine is unaffected by region and age, which can be widely applied in the whole country. An excellent immunization efficacy of the vaccine may be found in the HBV vaccine non - response crowd.%目的 探讨无应答人群接种60 ug重组乙型肝炎疫苗(酿酒酵母)免疫成功率和效果. 方法 选择桃江县40名17岁以上接种常规乙肝疫苗后抗体(抗-HBs)没有阳转的人群,应用深圳康泰生物制品股份有限公司生产的60 μg重组乙型肝炎疫苗(酿酒酵母),采用一针接种后1月采血,进行血清免疫学观察. 结果 接种人群均未出现局部反应和全身反应.接种一针后1个月,阳转率达97.5%,抗-HBs几何平均滴度(GMT)为50.657 mIU/ml.结论 60μg重组乙型肝炎疫苗(酿酒酵母)安全性良好,免疫成功率高,不受地域、年龄组等影响,可以在全国范围内广泛使用,对于常规免疫无应答人群使用60 μg/ml乙肝疫苗,免疫效果可能更好.

  16. RECOMBINANT INFLUENZA VACCINES

    OpenAIRE

    Sedova, E.; Shcherbinin, D.; Migunov, A.; Smirnov, Iu; Logunov, D.; Shmarov, M.; Tsybalova, L.; Naroditskiĭ, B.; O. Kiselev; Gintsburg, A.

    2012-01-01

    This review covers the problems encountered in the construction and production of new recombinant influenza vaccines. New approaches to the development of influenza vaccines are investigated; they include reverse genetics methods, production of virus-like particles, and DNA- and viral vector-based vaccines. Such approaches as the delivery of foreign genes by DNA- and viral vector-based vaccines can preserve the native structure of antigens. Adenoviral vectors are a promising gene-delivery pla...

  17. Studies on Immune Effect of Yeast-derived Recombinant Hepatitis B Vaccine%重组酵母乙型肝炎疫苗血清学免疫效果观察

    Institute of Scientific and Technical Information of China (English)

    尹桂成; 茅亚达; 王渤; 严加和; 刘亚华

    2001-01-01

    为探讨国产重组酵母乙型肝炎(乙肝)疫苗不同剂量、不同间隔时间免疫后的安全性和血清学免疫效果,在五总乡和兴东镇对乙肝表面抗原(HBsAg)阳性母亲和阴性母亲的新生儿,按第1针分别接种10μg或5μg乙肝疫苗,1个月后接种第2针(0.5μg),第3针(0.5μg)与第1针分别间隔5、6、8个月,观察免疫效果。结果显示,所有接种对象均未发现严重副反应。完成3针乙肝疫苗免疫后,HBsAg阴性母亲的新生儿第1针接种10μg组抗-HBs阳转率及抗体水平均高于5μg组,差别均有极显著的统计学意义。第3针乙肝疫苗与第3针百白破混合制剂(DPT)联合免疫,乙肝疫苗的免疫效果良好,DPT的免疫应答率及水平与非联合免疫的差别无显著的统计学意义。这表明国产重组酵母乙肝疫苗的安全性好,第1针10μg组免疫效果优于5μg组,与DPT的联合免疫是可行的。%The safety and immune effect of different dosages and intervals of domestic yeast-derived recombinant hepatitis B (rHB) vaccine were studied during the time of Dec.1988~Feb. 2000.88 infants born to HBsAg negative mothers were divided into two groups and gave them 3 rHB vaccine doses of 10-5-5μg respectively with a 0,1,6 schedule. 40 infants born to HBsAg positive mothers were treated in the same way. After full course immunization, the anti-HBs positive seroconversion rate and the GMT of the infants born to HBsAg negative mothers given 10μg rHB vaccine at the first dose were significantly higher than that in infants given 5μg at the first dose. These two indexs in infants born to HBsAg positive mothers given 10μg and 5μg respectively at first dose showed no significant difference statistically, but the HBsAg positive rate in 10μg group was 5.56% and in 5μg group was 13.64%, this indicated infants born to HBsAg positive mothers could not be well protected by increasing rHB vaccine dosage, they may be benefitted by

  18. Recombination reduction on lead halide perovskite solar cells based on low temperature synthesized hierarchical TiO2 nanorods

    Science.gov (United States)

    Jaramillo-Quintero, Oscar A.; Solís de La Fuente, Mauricio; Sanchez, Rafael S.; Recalde, Ileana B.; Juarez-Perez, Emilio J.; Rincón, Marina E.; Mora-Seró, Iván

    2016-03-01

    Intensive research on the electron transport material (ETM) has been pursued to improve the efficiency of perovskite solar cells (PSCs) and decrease their cost. More importantly, the role of the ETM layer is not yet fully understood, and research on new device architectures is still needed. Here, we report the use of three-dimensional (3D) TiO2 with a hierarchical architecture based on rutile nanorods (NR) as photoanode material for PSCs. The proposed hierarchical nanorod (HNR) films were synthesized by a two-step low temperature (180 °C) hydrothermal method, and consist of TiO2 nanorod trunks with optimal lengths of 540 nm and TiO2 nanobranches with lengths of 45 nm. Different device configurations were fabricated with TiO2 structures (compact layer, NR and HNR) and CH3NH3PbI3, using different synthetic routes, as the active material. PSCs based on HNR-CH3NH3PbI3 achieved the highest power conversion efficiency compared to PSCs with other TiO2 structures. This result can be ascribed mainly to lower charge recombination as determined by impedance spectroscopy. Furthermore, we have observed that the CH3NH3PbI3 perovskite deposited by the two-step route shows higher efficiency, surface coverage and infiltration within the structure of 3D HNR than the one-step CH3NH3PbI3-xClx perovskite.Intensive research on the electron transport material (ETM) has been pursued to improve the efficiency of perovskite solar cells (PSCs) and decrease their cost. More importantly, the role of the ETM layer is not yet fully understood, and research on new device architectures is still needed. Here, we report the use of three-dimensional (3D) TiO2 with a hierarchical architecture based on rutile nanorods (NR) as photoanode material for PSCs. The proposed hierarchical nanorod (HNR) films were synthesized by a two-step low temperature (180 °C) hydrothermal method, and consist of TiO2 nanorod trunks with optimal lengths of 540 nm and TiO2 nanobranches with lengths of 45 nm. Different

  19. Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination.

    Directory of Open Access Journals (Sweden)

    Sjoerd H E van den Worm

    Full Text Available Severe acute respiratory syndrome (SARS is a zoonotic disease caused by SARS-related coronavirus (SARS-CoV that emerged in 2002 to become a global health concern. Although the original outbreak was controlled by classical public health measures, there is a real risk that another SARS-CoV could re-emerge from its natural reservoir, either in its original form or as a more virulent or pathogenic strain; in which case, the virus would be difficult to control in the absence of any effective antiviral drugs or vaccines. Using the well-studied SARS-CoV isolate HKU-39849, we developed a vaccinia virus-based SARS-CoV reverse genetic system that is both robust and biosafe. The SARS-CoV genome was cloned in separate vaccinia virus vectors, (vSARS-CoV-5prime and vSARS-CoV-3prime as two cDNAs that were subsequently ligated to create a genome-length SARS-CoV cDNA template for in vitro transcription of SARS-CoV infectious RNA transcripts. Transfection of the RNA transcripts into permissive cells led to the recovery of infectious virus (recSARS-CoV. Characterization of the plaques produced by recSARS-CoV showed that they were similar in size to the parental SARS-CoV isolate HKU-39849 but smaller than the SARS-CoV isolate Frankfurt-1. Comparative analysis of replication kinetics showed that the kinetics of recSARS-CoV replication are similar to those of SARS-CoV Frankfurt-1, although the titers of virus released into the culture supernatant are approximately 10-fold less. The reverse genetic system was finally used to generate a recSARS-CoV reporter virus expressing Renilla luciferase in order to facilitate the analysis of SARS-CoV gene expression in human dendritic cells (hDCs. In parallel, a Renilla luciferase gene was also inserted into the genome of human coronavirus 229E (HCoV-229E. Using this approach, we demonstrate that, in contrast to HCoV-229E, SARS-CoV is not able to mediate efficient heterologous gene expression in hDCs.

  20. Transmission of hepatitis B in Hamburg, Germany, 1998-2002: a prospective, population-based study.

    Science.gov (United States)

    Diel, R; Helle, J; Gottschalk, R

    2005-08-01

    To study the pattern of transmission of HBV in a large urban community, an in-depth prospective study was performed in Hamburg between 1 January 1998 and 31 December 2002. In total, 524 patients were classified as hepatitis B cases according to the case definition of the Robert Koch Institute, comprising 197 foreign-born and 327 German-born persons. The principal risk factor was parenteral drug use, with 17.7% (n=93/524) of all documented cases of hepatitis B, followed by immigration as refugees (13.9%; n=73). Of all 524 cases, 72 (13.7%) were associated with heterosexual (n=41) or homosexual (n=31) transmission. Household contacts of HBV carriers or of patients with acute infectious disease contributed to 9.0% of the cases (n=47). Medical procedures were most probably the source in 7.4% (n=39), although only 3.2% (n=17) of all patients were health-care workers. In multivariate analysis of household contacts, male-male sexual activity was found to be the greatest risk factor for acquiring an acute HBV infection, followed by asylum-seeking status and the number of contacts. The incidence was 3.5-fold higher among foreign-born persons (16.1 per 100,000) than among German-born individuals (4.5 per 100,000) suggesting that a targeted intervention in this population group is a public-health need. The current national policy of vaccination in defined age groups should be extended to the immunization of all children of foreign-born parents as well as the screening and immunisation of susceptible foreign-born adults.

  1. Predicting the expression of recombinant monoclonal antibodies in Chinese hamster ovary cells based on sequence features of the CDR3 domain.

    Science.gov (United States)

    Pybus, Leon P; James, David C; Dean, Greg; Slidel, Tim; Hardman, Colin; Smith, Andrew; Daramola, Olalekan; Field, Ray

    2014-01-01

    Despite the development of high-titer bioprocesses capable of producing >10 g L(-1) of recombinant monoclonal antibody (MAb), some so called "difficult-to-express" (DTE) MAbs only reach much lower process titers. For widely utilized "platform" processes the only discrete variable is the protein coding sequence of the recombinant product. However, there has been little systematic study to identify the sequence parameters that affect expression. This information is vital, as it would allow us to rationally design genetic sequence and engineering strategies for optimal bioprocessing. We have therefore developed a new computational tool that enables prediction of MAb titer in Chinese hamster ovary (CHO) cells based on the recombinant coding sequence of the expressed MAb. Model construction utilized a panel of MAbs, which following a 10-day fed-batch transient production process varied in titer 5.6-fold, allowing analysis of the sequence features that impact expression over a range of high and low MAb productivity. The model identified 18 light chain (LC)-specific sequence features within complementarity determining region 3 (CDR3) capable of predicting MAb titer with a root mean square error of 0.585 relative expression units. Furthermore, we identify that CDR3 variation influences the rate of LC-HC dimerization during MAb synthesis, which could be exploited to improve the production of DTE MAb variants via increasing the transfected LC:HC gene ratio. Taken together these data suggest that engineering intervention strategies to improve the expression of DTE recombinant products can be rationally implemented based on an identification of the sequence motifs that render a recombinant product DTE.

  2. Hepatic Encephalopathy

    Medline Plus

    Full Text Available ... is a condition that causes temporary worsening of brain function in people with advanced liver disease. When ... travel through your body until they reach your brain, causing mental and physical symptoms of HE. Hepatic ...

  3. Hepatic Encephalopathy

    Medline Plus

    Full Text Available ... Hepatic Encephalopathy so you can tell your doctor right away if you think you may have it. ... American Liver Foundation © 2016 American Liver Foundation. All rights reserved. Funding for the HE123 - Diagnosis, Treatment and ...

  4. Hepatic Encephalopathy

    Medline Plus

    Full Text Available ... Symptoms to look for Caregiver Support Caregiver Stories Home › What is Hepatic Encephalopathy? Why Your Liver is ... questions about HE, one step at a time. Home About Us Ways to Give Contact Us Privacy ...

  5. Autoimmune hepatitis

    Science.gov (United States)

    ... Sjogren syndrome Systemic lupus erythematosus Thyroiditis Type 1 diabetes Ulcerative colitis Autoimmune hepatitis may occur in family members of people with autoimmune diseases. There may be a genetic cause. This disease is most common in young girls ...

  6. Hepatic encephalopathy

    Science.gov (United States)

    ... mild and include: Breath with a musty or sweet odor Change in sleep patterns Changes in thinking ... 24411831 . Nevah MI, Fallon MB. Hepatic encephalopathy, hepatorenal syndrome, hepatopumonary syndrome, and systemic complications of liver disease. ...

  7. Hepatic Encephalopathy

    Medline Plus

    Full Text Available ... Get Worse? How is HE Diagnosed? Prior to Treatment Who treats HE? Preparing for your Medical Appointment Hepatic Encephalopathy Treatment Options Treatment Basics Treatment Medications Importance of Adhering ...

  8. Hepatitis B

    Science.gov (United States)

    ... and Indian subcontinent) Reside or work in a prison or correctional facility People with end-stage renal ... to monitor and reduce their drinking behavior. Glossary Definitions of terms commonly used with viral hepatitis and ...

  9. Recombination with coat protein transgene in a complemen-tation system based on Cucumber mosaic virus (CMV)

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    In order to study the feasibility of Cucumber mosaic virus (CMV) as an expression vector, the full-length cDNA of RNA 3 from strain SD was cloned and the sequence around the start codon of the coat protein (CP) gene was modified to create an NsiⅠ site for insertion of foreign genes. The CP gene was replaced by the green fluorescent protein (GFP) gene. The cDNAs of Fny RNAs 1 and 2 and the chimeric SD RNA 3 were cloned between the modified 35S promoter and terminator. Tobacco protoplasts were transfected with a mixture of the viral cDNAs containing 35S promoter and terminator as a replacement vector and expressed GFP. A complementation system was established when the replacement vector was inoculated onto the transgenic tobacco plants ex-pressing SD-CMV CP. GFP was detected in the inoculated leaves in 5 of 18 tested plants and in the first upper systemic leaf of one of the 5 plants ten days after inoculation. However, no GFP could be detected in all the plants one month after inoculation. Recombination between the CMV vector and the CP transgene was proved by retro-transcriptional polymerase chain reaction (RT-PCR) and verified by DNA sequencing. Our results argue against the feasibility of the CMV-based replace-ment vector trans-complemented by the CP transgene, and at the same time, enlighten ways to improve the CMV-based expression vector and the biosafety of CMV CP-mediated virus resistant transgenic plants.

  10. Diagnostic criteria of autoimmune hepatitis.

    Science.gov (United States)

    Liberal, Rodrigo; Grant, Charlotte R; Longhi, Maria Serena; Mieli-Vergani, Giorgina; Vergani, Diego

    2014-01-01

    Autoimmune hepatitis (AIH) is a chronic immune-mediated liver disorder characterised by female preponderance, elevated transaminase and immunoglobulin G levels, seropositivity for autoantibodies and interface hepatitis. Presentation is highly variable, therefore AIH should be considered during the diagnostic workup of any increase in liver enzyme levels. A set of inclusion and exclusion criteria for the diagnosis of AIH have been established by the International Autoimmune Hepatitis Group (IAIHG). There are two main types of AIH: type 1, positive for anti-nuclear (ANA) and/or anti-smooth muscle antibodies (SMAs) and type 2, defined by the presence of anti-liver kidney microsomal antibody type 1 (LKM-1) and/or anti-liver cytosol type 1 (LC-1) autoantibodies. The central role of autoantibodies in the diagnosis of AIH has led the IAIHG to produce a consensus statement detailing appropriate and effective methods for their detection. Autoantibodies should be tested by indirect immunofluorescence at an initial dilution of 1/40 in adults and 1/10 in children on a freshly prepared rodent substrate that includes kidney, liver and stomach sections to allow for the simultaneous detection of all reactivities relevant to AIH. Anti-LKM-1 is often confused with anti-mitochondrial antibody (AMA) if rodent kidney is used as the sole immunofluorescence substrate. The identification of the molecular targets of anti-LKM-1 and AMA has led to the establishment of immuno-assays based on the use of the recombinant or purified autoantigens. Perinuclear anti-nuclear neutrophil antibody (p-ANNA) is an additional marker of AIH-1; anti soluble liver antigen (SLA) antibodies are specific for autoimmune liver disease, can be present in AIH-1 and AIH-2 and are associated with a more severe clinical course. Anti-SLA are detectable by ELISA or radio-immuno-assays, but not by immunofluorescence. AIH is exquisitely responsive to immunosuppressive treatment, which should be instituted promptly to

  11. Hepatitis C and Incarceration

    Science.gov (United States)

    ... It is also the most common type in jails and prisons. What is Hepatitis C? Hepatitis C is a ... risk for Hepatitis C because many people in jails or prisons already have Hepatitis C. • The most ...

  12. Hepatitis A Test

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Hepatitis A Testing Share this page: Was this page ... HAV-Ab total; Anti-HAV Formal name: Viral Hepatitis A Antibody Related tests: Hepatitis B Testing ; Hepatitis ...

  13. Hepatitis A FAQs

    Science.gov (United States)

    ... of Viral Hepatitis Contact Us Quick Links to Hepatitis ... A | B | C | D | E Viral Hepatitis Home ... Outbreaks State and Local Partners & Grantees Resource Center Hepatitis A FAQs for the Public Recommend on Facebook ...

  14. Protect Yourself from Hepatitis

    Science.gov (United States)

    ... Your Liver Guard Your Liver Protect Yourself From Hepatitis Hepatitis can make you feel as if you have ... viruses that attack your lungs and respiratory system; hepatitis is a liver disease. Some forms of hepatitis ...

  15. Delta agent (Hepatitis D)

    Science.gov (United States)

    Hepatitis D virus ... Hepatitis D virus (HDV) is found only in people who carry the hepatitis B virus. HDV may make liver ... B virus but who never had symptoms. Hepatitis D infects about 15 million people worldwide. It occurs ...

  16. Hepatitis B Foundation

    Science.gov (United States)

    ... worldwide 2 Billion People have been infected with Hepatitis B Worldwide The Hepatitis B Foundation is working ... of people living with hepatitis B. Learn About Hepatitis B in 10 Other Languages . Resource Video See ...

  17. Dose-reduced CT with model-based iterative reconstruction in evaluations of hepatic steatosis: How low can we go?

    Energy Technology Data Exchange (ETDEWEB)

    Yasaka, Koichiro, E-mail: koyasaka@gmail.com [Department of Radiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan); Katsura, Masaki [Department of Radiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan); Akahane, Masaaki [NTT Medical Center Tokyo, 5-9-22 Higashi-Gotanda, Shinagawa-ku, Tokyo 141-8625 (Japan); Sato, Jiro [Department of Radiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan); Matsuda, Izuru [Kanto Rosai Hospital, 1-1 Kizukisumiyoshi-cho, Nakahara-ku, Kawasaki, Kanagawa 211-8510 (Japan); Ohtomo, Kuni [Department of Radiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan)

    2014-07-15

    Purpose: To determine whether dose-reduced CT with model-based iterative image reconstruction (MBIR) is a useful tool with which to diagnose hepatic steatosis. Materials and methods: This prospective clinical study approved by our Institutional Review Board included 103 (67 men and 36 women; mean age, 64.3 years) patients who provided written informed consent to undergo unenhanced CT. Images of reference-dose CT (RDCT) with filtered back projection (R-FBP) and low- and ultralow-dose CT (dose-length product; 24 and 9% of that of RDCT) with MBIR (L-MBIR and UL-MBIR) were reconstructed. Mean CT numbers of liver (CT[L]) and spleen (CT[S]), and quotient (CT[L/S]) of CT[L] and CT[S] were calculated from selected regions of interest. Bias and limits of agreement (LOA) of CT[L] and CT[L/S] in L-MBIR and UL-MBIR (vs. R-FBP) were assessed using Bland–Altman analyses. Diagnostic methods for hepatic steatosis of CT[L] < 48 Hounsfield units (HU) and CT[L/S] < 1.1 were applied to L-MBIR and UL-MBIR using R-FBP as the reference standard. Results: Bias was larger for CT[L] in UL-MBIR than in L-MBIR (−3.18 HU vs. −1.73 HU). The LOA of CT[L/S] was larger for UL-MBIR than for L-MBIR (±0.425 vs. ±0.245) and outliers were identified in CT[L/S] of UL-MBIR. Accuracy (0.92–0.95) and the area under the receiver operating characteristics curve (0.976–0.992) were high for each method, but some were slightly lower in UL-MBIR than L-MBIR. Conclusion: Dose-reduced CT reconstructed with MBIR is applicable to diagnose hepatic steatosis, however, a low dose of radiation might be preferable.

  18. Feature Hepatitis: Hepatitis Symptoms, Diagnosis, Treatment & Prevention

    Science.gov (United States)

    ... Navigation Bar Home Current Issue Past Issues Feature Hepatitis Hepatitis: Symptoms, Diagnosis, Treatment & Prevention Past Issues / Spring 2009 ... No appetite Fever Headaches Diagnosis To check for hepatitis viruses, your doctor will test your blood. You ...

  19. A recombinant Hendra virus G glycoprotein-based subunit vaccine protects ferrets from lethal Hendra virus challenge.

    Science.gov (United States)

    Pallister, Jackie; Middleton, Deborah; Wang, Lin-Fa; Klein, Reuben; Haining, Jessica; Robinson, Rachel; Yamada, Manabu; White, John; Payne, Jean; Feng, Yan-Ru; Chan, Yee-Peng; Broder, Christopher C

    2011-08-01

    The henipaviruses, Hendra virus (HeV) and Nipah virus (NiV), are two deadly zoonotic viruses for which no vaccines or therapeutics have yet been approved for human or livestock use. In 14 outbreaks since 1994 HeV has been responsible for multiple fatalities in horses and humans, with all known human infections resulting from close contact with infected horses. A vaccine that prevents virus shedding in infected horses could interrupt the chain of transmission to humans and therefore prevent HeV disease in both. Here we characterise HeV infection in a ferret model and show that it closely mirrors the disease seen in humans and horses with induction of systemic vasculitis, including involvement of the pulmonary and central nervous systems. This model of HeV infection in the ferret was used to assess the immunogenicity and protective efficacy of a subunit vaccine based on a recombinant soluble version of the HeV attachment glycoprotein G (HeVsG), adjuvanted with CpG. We report that ferrets vaccinated with a 100 μg, 20 μg or 4 μg dose of HeVsG remained free of clinical signs of HeV infection following a challenge with 5000 TCID₅₀ of HeV. In addition, and of considerable importance, no evidence of virus or viral genome was detected in any tissues or body fluids in any ferret in the 100 and 20 μg groups, while genome was detected in the nasal washes only of one animal in the 4 μg group. Together, our findings indicate that 100 μg or 20 μg doses of HeVsG vaccine can completely prevent a productive HeV infection in the ferret, suggesting that vaccination to prevent the infection and shedding of HeV is possible.

  20. Chromogenic platform based on recombinant Drosophila melanogaster acetylcholinesterase for visible unidirectional assay of organophosphate and carbamate insecticide residues

    Energy Technology Data Exchange (ETDEWEB)

    Han Zheng [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Chi Chensen [School of Life Science and Biotechnology, Bor Luh Food Safety Center, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Bai Bing; Liu Gang; Rao Qinxiong [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Peng Shaojie [Institute of Shanghai Food and Drug Supervision, 615 Liuzhou Road, Shanghai 200233 (China); Liu Hong [Shanghai Municipal Center for Disease Control and Prevention, 1380 Zhongshan West Road, Shanghai 200336 (China); Zhao Zhihui [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Zhang Dabing [School of Life Science and Biotechnology, Bor Luh Food Safety Center, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Wu Aibo, E-mail: wuaibo@saas.sh.cn [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China)

    2012-03-30

    Highlight: Black-Right-Pointing-Pointer A visible chromogenic platform for rapid analysis of OP and CM insecticide residues was developed. Black-Right-Pointing-Pointer The assay has the capabilities of both qualitative measurement and quantitative analysis. Black-Right-Pointing-Pointer The sensitivity, capabilities of resisting interferences and storage stability were desirable. Black-Right-Pointing-Pointer Matrix effects were acceptable and detection performance was satisfactory in real application. - Abstract: In this study we propose a chromogenic platform for rapid analysis of organophosphate (OP) and carbamate (CM) insecticide residues, based on recombinant Drosophila melanogaster acetylcholinesterase (R-DmAChE) as enzyme and indoxyl acetate as substrate. The visible chromogenic strip had the advantages identical to those of commonly used lateral flow assays (LFAs) with utmost simplicity in sample loading and result observation. After optimization, depending on the color intensity (CI) values, the well-established assay has the capabilities of both qualitative measurement via naked eyes and quantitative analysis by colorimetric reader with the desirable IC{sub 50} values against the tested six insecticides (0.06 {mu}g mL{sup -1} of carbofuran, 0.28 {mu}g mL{sup -1} of methomyl, 0.03 {mu}g mL{sup -1} of dichlorvos, 31.6 {mu}g mL{sup -1} of methamidophos, 2.0 {mu}g mL{sup -1} of monocrotophos, 6.3 {mu}g mL{sup -1} of omethoate). Acceptable matrix effects and satisfactory detection performance were confirmed by in-parallel LC-MS/MS analysis in different vegetable varieties at various spiked levels of 10{sup -3} to 10{sup 1} {mu}g g{sup -1}. Overall, the testified suitability and applicability of this novel platform meet the requirements for practical use in food safety management and environmental monitoring, especially in the developing world.

  1. Vaccine development for allergen-specific immunotherapy based on recombinant allergens and synthetic allergen peptides: Lessons from the past and novel mechanisms of action for the future

    OpenAIRE

    Valenta, Rudolf; Campana, Raffaela; Focke-Tejkl, Margit; Niederberger, Verena

    2016-01-01

    In the past, the development of more effective, safe, convenient, broadly applicable, and easy to manufacture vaccines for allergen-specific immunotherapy (AIT) has been limited by the poor quality of natural allergen extracts. Progress made in the field of molecular allergen characterization has now made it possible to produce defined vaccines for AIT and eventually for preventive allergy vaccination based on recombinant DNA technology and synthetic peptide chemistry. Here we review the char...

  2. Degree of tumour vascularity correlates with drug accumulation and tumour response upon TNF-α-based isolated hepatic perfusion

    NARCIS (Netherlands)

    B. van Etten (Boudewijn); M.R. de Vries (Mark); M.G.A. van IJken (Marc); T. Lans (Titia); G. Guetens (Gunther); F. Ambagtsheer (Frederike); S.T. van Tiel (Sandra); G. de Boeck (Gert); E.A. de Bruijn (Ernst); A.M.M. Eggermont (Alexander); T.L.M. ten Hagen (Timo)

    2003-01-01

    textabstractIsolated hepatic perfusion (IHP) with melphalan with or without tumour necrosis factor alpha (TNF-α) is currently performed in clinical trials in patients with hepatic metastases. Previous studies led to the hypothesis that the use of TNF-α in isolated limb perfusion causes specific dest

  3. How to Treat Pain in the Hepatic Region Due to Chronic Hepatitis?

    Institute of Scientific and Technical Information of China (English)

    林宗广

    2004-01-01

    @@ Chronic viral hepatitis type B and C both have the symptoms of pain in the hepatic region, asthenia, poor appetite, abdominal fullness, among which pain in the hepatic region is the most commonly seen. According to the author's clinical experience, treatment based on accurate TCM differentiation can not only eliminate pain in the hepatic region but also restore the hepatic function at the same time. Differentiation includes analysis of the nature of the hepatic pain and the accompanying symptoms, and the treatment is aimed at the differentiated symptoms. The following are methods of treatment.

  4. Importance of virological response in the early stage of telaprevir-based triple therapy for hepatitis C

    Institute of Scientific and Technical Information of China (English)

    Satoshi; Hiramine; Norihiro; Furusyo; Eiichi; Ogawa; Makoto; Nakamuta; Eiji; Kajiwara; Hideyuki; Nomura; Kazufumi; Dohmen; Kazuhiro; Takahashi; Takeaki; Satoh; Koichi; Azuma; Akira; Kawano; Toshimasa; Koyanagi; Kazuhiro; Kotoh; Shinji; Shimoda; Jun; Hayashi

    2015-01-01

    AIM: To investigate the efficacy of virological response(VR) to telaprevir(TVR)-based triple therapy in predicting treatment outcome of hepatitis C.METHODS: This prospective, multicenter study consisted of 253 Japanese patients infected with hepatitis C virus(HCV) genotype 1b. All received 12 wk of TVR in combination with 24 wk of pegylatedinterferon-α(IFN-α) and ribavirin. Serum HCV RNA was tested at weeks 1, 2, 3, 4, 6, 8, 12, 16, 20, and 24. VR was defined as undetectable serum HCV RNA. Sustained virological response(SVR) was VR at 24 wk after the end of treatment and was regarded as a successful outcome.RESULTS: Of 253 patients, 207(81.8%) achieved SVR. The positive predictive value of VR for SVR was 100% at week 2, after which it gradually decreased, and was over 85% to week 12. The negative predictive value(NPV) gradually increased, reaching 100% at week 12. The upslope of the NPV showed a large increase from week 4(40.6%) to week 6(82.4%). There was a moderate concordance between the SVR and VR at week 6(kappa coefficient = 0.44), although other VRs had poor concordance to SVR. Multiple logistic regression analysis extracted VR at week 6(P < 0.0001, OR = 63.8) as an independent factor contributing to SVR. In addition, the interleukin-28 B single nucleotide polymorphism and response to previous pegylated-IFN-α and ribavirin therapy were identified as independent factors for SVR.CONCLUSION: VR at week 6, but not at week 4, is an efficient predictor of both SVR and non-SVR to TVRbased triple therapy.

  5. Recombinant HCV variants with NS5A from genotypes 1-7 have different sensitivities to an NS5A inhibitor but not interferon-a

    DEFF Research Database (Denmark)

    Scheel, Troels K H; Gottwein, Judith M; Mikkelsen, Lotte S;

    2011-01-01

    Heterogeneity in the hepatitis C virus (HCV) protein NS5A influences its sensitivity to interferon-based therapy. Furthermore, NS5A is an important target for development of HCV-specific inhibitors. We aimed to develop recombinant infectious cell culture systems that express NS5A from isolates...... of the 7 major HCV genotypes, and determining their sensitivity to a specific NS5A inhibitor and to interferon-a....

  6. Noninvasive diagnosis of hepatic fibrosis in chronic hepatitis C

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Assessment of hepatic fibrosis is important for determining prognosis, guiding management decisions,and monitoring disease. Histological evaluation of liver biopsy specimens is currently considered the reference test for staging hepatic fibrosis. Since liver biopsy carries a small but significant risk, noninvasive tests to assess hepatic fibrosis are desirable. This editorial gives an overview on noninvasive methods currently available to determine hepatic fibrosis and their diagnostic accuracy for predicting significant fibrosis and cirrhosis in chronic hepatitis C. Based on available data, the performance of simple tests derived from routine laboratory parameters appears to be similar to that of more complex and expensive fibrosis panels. Transient elastography seems more accurate than blood tests for diagnosing cirrhosis.

  7. Stoichiometry Based Steady-State Hepatic Flux Analysis: Computational and Experimental Aspects

    Directory of Open Access Journals (Sweden)

    Mehmet A. Orman

    2012-03-01

    Full Text Available The liver has many complex physiological functions, including lipid, protein and carbohydrate metabolism, as well as bile and urea production. It detoxifies toxic substances and medicinal products. It also plays a key role in the onset and maintenance of abnormal metabolic patterns associated with various disease states, such as burns, infections and major traumas. Liver cells have been commonly used in in vitro experiments to elucidate the toxic effects of drugs and metabolic changes caused by aberrant metabolic conditions, and to improve the functions of existing systems, such as bioartificial liver. More recently, isolated liver perfusion systems have been increasingly used to characterize intrinsic metabolic changes in the liver caused by various perturbations, including systemic injury, hepatotoxin exposure and warm ischemia. Metabolic engineering tools have been widely applied to these systems to identify metabolic flux distributions using metabolic flux analysis or flux balance analysis and to characterize the topology of the networks using metabolic pathway analysis. In this context, hepatic metabolic models, together with experimental methodologies where hepatocytes or perfused livers are mainly investigated, are described in detail in this review. The challenges and opportunities are also discussed extensively.

  8. Hepatitis C Virus Protein Interaction Network Analysis Based on Hepatocellular Carcinoma.

    Science.gov (United States)

    Han, Yuewen; Niu, Jun; Wang, Dong; Li, Yuanyuan

    2016-01-01

    Epidemiological studies have validated the association between hepatitis C virus (HCV) infection and hepatocellular carcinoma (HCC). An increasing number of studies show that protein-protein interactions (PPIs) between HCV proteins and host proteins play a vital role in infection and mediate HCC progression. In this work, we collected all published interaction between HCV and human proteins, which include 455 unique human proteins participating in 524 HCV-human interactions. Then, we construct the HCV-human and HCV-HCC protein interaction networks, which display the biological knowledge regarding the mechanism of HCV pathogenesis, particularly with respect to pathogenesis of HCC. Through in-depth analysis of the HCV-HCC interaction network, we found that interactors are enriched in the JAK/STAT, p53, MAPK, TNF, Wnt, and cell cycle pathways. Using a random walk with restart algorithm, we predicted the importance of each protein in the HCV-HCC network and found that AKT1 may play a key role in the HCC progression. Moreover, we found that NS5A promotes HCC cells proliferation and metastasis by activating AKT/GSK3β/β-catenin pathway. This work provides a basis for a detailed map tracking new cellular interactions of HCV and identifying potential targets for HCV-related hepatocellular carcinoma treatment. PMID:27115606

  9. Community-based cross-sectional seroprevalence study of hepatitis A in Bangladesh

    Institute of Scientific and Technical Information of China (English)

    Samir K Saha; Setarunnahar Saha; Salim Shakur; Mohammed Hanif; Md Ahsan Habib; Sanjoy K Datta; Hans L Bock

    2009-01-01

    AIM: To elucidate the age-distribution of anti-hepatitis A virus (HAV) seroprevalence across different socioeconomic status (SES) categories in Bangladesh which,despite scarce data, is generally deemed to have high endemicity.METHODS: Blood samples of 818 subjects from a strati-fied sample of schools and hospitals, comprising different age categories and SES were collected. They were assayed for total anti-HAV antibodies. Social and medical history data were obtained using a questionnaire.RESULTS: Overall anti-HAV seroprevalence was 69.6%, increasing with age from 1-5 years (40.4%) to > 30 years (98.4%). Seroprevalence was lowest (49.8%) in the high SES group and highest (96.5%) in the rural lower-middle SES group. Among subjects aged 6-20 years, anti-HAV seroprevalence was lowest in urban private school children (43.0%), followed by urban government school children (76.2%) and rural school children (96.5%) ( P < 0.01). Within the high SES group, anti-HAV seroprevalence was 32.3% in subjects < 10 years and 51.7% in those aged 11-20 years. Until now Bangladesh has been deemed to have high endemicity for HAV.CONCLUSION: The transition from high to intermediate HAV endemicity may be underway; high SES adolescents and adults remain particularly at risk of symptomatic illness. Preventive measures need consideration.

  10. Relation Between Hepatitis C Virus Exposure and Risk of Osteoporosis: A Nationwide Population-Based Study.

    Science.gov (United States)

    Chen, Chien-Hua; Lin, Cheng-Li; Kao, Chia-Hung

    2015-11-01

    The effect of hepatitis C virus (HCV) exposure on bone mineral density without advanced liver disease remains debated. Thus, we assessed the relation between HCV exposure and the risk of osteoporosis.From 2000 to 2011, patients aged >20 years with HCV exposure were identified from the Longitudinal Health Insurance Database 2000. Of the 51,535 sampled patients, 41,228 and 10,307 patients were categorized as the comparison and the HCV exposure cohorts, respectively.The overall incidence of osteoporosis in the HCV exposure cohort was higher than in the comparison cohort (8.27 vs 6.19 per 1000 person-years; crude hazard ratio = 1.33, 95% confidence interval = 1.20-1.47). The incidence of osteoporosis, higher in women than in men, increased with age and comorbidity of hypertension, hyperlipidemia, and heart failure. The risk of developing osteoporosis was significantly higher in the HCV exposure cohort than in the comparison cohort after adjusting for age, sex, diabetes, hypertension, hyperlipidemia, heart failure, stroke, and cirrhosis. However, the risk of osteoporosis contributed by HCV decreased with age and the presence of comorbidity. Furthermore, the risk of osteoporotic fracture did not differ significantly between patients exposed to HCV and the comparison cohorts.HCV increases the risk of osteoporosis, but no detrimental effect on osteoporotic fracture was observed in this study. Furthermore, HCV may be less influential than other risk factors, such as hypertension, hyperlipidemia, and heart failure, in contributing to the development of osteoporosis. PMID:26632720

  11. Hepatitis C Virus Protein Interaction Network Analysis Based on Hepatocellular Carcinoma.

    Directory of Open Access Journals (Sweden)

    Yuewen Han

    Full Text Available Epidemiological studies have validated the association between hepatitis C virus (HCV infection and hepatocellular carcinoma (HCC. An increasing number of studies show that protein-protein interactions (PPIs between HCV proteins and host proteins play a vital role in infection and mediate HCC progression. In this work, we collected all published interaction between HCV and human proteins, which include 455 unique human proteins participating in 524 HCV-human interactions. Then, we construct the HCV-human and HCV-HCC protein interaction networks, which display the biological knowledge regarding the mechanism of HCV pathogenesis, particularly with respect to pathogenesis of HCC. Through in-depth analysis of the HCV-HCC interaction network, we found that interactors are enriched in the JAK/STAT, p53, MAPK, TNF, Wnt, and cell cycle pathways. Using a random walk with restart algorithm, we predicted the importance of each protein in the HCV-HCC network and found that AKT1 may play a key role in the HCC progression. Moreover, we found that NS5A promotes HCC cells proliferation and metastasis by activating AKT/GSK3β/β-catenin pathway. This work provides a basis for a detailed map tracking new cellular interactions of HCV and identifying potential targets for HCV-related hepatocellular carcinoma treatment.

  12. Voxel-based analyses of magnetization transfer imaging of the brain in hepatic encephalopathy

    Institute of Scientific and Technical Information of China (English)

    Falk R Miese; Hans-J(o)rg Wittsack; Gerald Kircheis; Arne Holstein; Christian Mathys; Ulrich M(o)dder; Mathias Cohnen

    2009-01-01

    AIM: To evaluate the spatial distribution of cerebral abnormalities in cirrhotic subjects with and without hepatic encephalopathy (HE) found with magnetization transfer imaging (MTI). METHODS: Nineteen cirrhotic patients graded from neurologically normal to HE grade 2 and 18 healthy control subjects underwent magnetic resonance imaging. They gave institutional-review-board-approved written consent. Magnetization transfer ratio (MTR) maps were generated from MTI. We tested for significant differences compared to the control group using statistical non-parametric mapping (SnPM) for a voxelbased evaluation. RESULTS: The MTR of grey and white matter was lower in subjects with more severe HE. Changes were found in patients with cirrhosis without neurological deficits in the basal ganglia and bilateral white matter. The loss in magnetization transfer increased in severity and spatial extent in patients with overt HE. Patients with HE grade 2 showed an MTR decrease in white and grey matter: the maximum loss of magnetization transfer effect was located in the basal ganglia [SnPM (pseudo-)t = 17.98, P = 0.0001]. CONCLUSION: The distribution of MTR changes in HE points to an early involvement of basal ganglia and white matter in HE.

  13. Chronic hepatitis C is a common associated with hepatic granulomas

    Institute of Scientific and Technical Information of China (English)

    Ned Snyder; Juan G Martinez; Shu-Yuan Xiao

    2008-01-01

    AIM: To determine the most frequent etiologies of hepatic epithelioid granulomas, and whether there was an association with chronic hepatitis C virus (HCV). METHODS: Both a retrospective review of the pathol-ogy database of liver biopsies at our institution from 1996 through 2006 as well as data from a prospective study of hepatic fibrosis markers and liver biopsies from 2003 to 2006 were reviewed to identify cases of hepatic epithelioid granulomas. Appropriate charts, liver biopsy slides, and laboratory data were reviewed to determine all possible associations. The diagnosis of HCV was based on a positive HCV RNA. RESULTS: There were 4578 liver biopsies and 36 (0.79%) had at least one epithelioid granuloma. HCV was the most common association. Fourteen patients had HCV, and in nine, there were no concurrent condi-tions known to be associated with hepatic granulomas. Prior interferon therapy and crystalloid substances from illicit intravenous injections did not account for the finding. There were hepatic epithelioid granulomas in 3 of 241 patients (1.24%) with known chronic HCV enrolled in the prospective study of hepatic fibrosis markers. CONCLUSION: Although uncommon, hepatic granu-Iomas may be part of the histological spectrum of chronic HCV. When epithelioid granulomas are found on the liver biopsy of someone with HCV, other clini-cally appropriate studies should be done, but if nothing else is found, the clinician can be comfortable with an HCV association.

  14. Immunogenicity and safety of hepatitis E vaccine in healthy hepatitis B surface antigen positive adults

    OpenAIRE

    Wu, Ting; Huang, Shou-Jie; Zhu, Feng-Cai; Zhang, Xue-Feng; AI, XING; Yan, Qiang; Wang, Zhong-Ze; Yang, Chang-Lin; Jiang, Han-Min; Liu, Xiao-Hui; Guo, Meng; Du, Hai-Lian; Ng, Mun-Hon; Zhang, Jun; Xia, Ningshao

    2013-01-01

    A recombinant hepatitis E vaccine, Hecolin®, has been proven safe and effective in healthy adults. As hepatitis B surface antigen (HBsAg) positive individuals have a higher risk of poor prognosis after super-infection with hepatitis E virus (HEV), the safety and immunogenicity of Hecolin® in this population should be assessed. The present study is an extending analysis of data from a large randomized controlled clinical trial of Hecolin®. Healthy participants (n = 14,065) without current or p...

  15. High-resolution genetic maps of Lotus japonicus and L. burttii based on re-sequencing of recombinant inbred lines

    DEFF Research Database (Denmark)

    Shah, Niraj; Hirakawa, Hideki; Kusakabe, Shohei;

    2016-01-01

    Recombinant inbred lines (RILs) derived from bi-parental populations are stable genetic resources, which are widely used for constructing genetic linkage maps. These genetic maps are essential for QTL mapping and can aid contig and scaffold anchoring in the final stages of genome assembly. In thi...

  16. Full protection in mink against mink enteritis virus with new generation canine parvovirus vaccines based on synthetic peptide or recombinant protein

    DEFF Research Database (Denmark)

    Langeveld, J. P.; Kamstrup, Søren; Uttenthal, Åse;

    1995-01-01

    Two recently developed vaccines—one based on synthetic peptide and one based on recombinant capsid protein—fully protected dogs against heavy experimental canine parvovirus (CPV) infection. The high sequence homology (>98%) and antigenic similarity between CPV and mink enteritis virus (MEV), feline...... panleukopenia virus, and raccoon parvovirus, suggest that both vaccines could protect mink, cats and raccoons against these respective host range variants. This was tested in mink and turned out to be the case. The two vaccines were fully protective and as effective as a conventional commercial vaccine based...

  17. Improvement of the Chromosome-based Recombineering System%基于染色体的重组工程系统的改进

    Institute of Scientific and Technical Information of China (English)

    樊丹; 石牡丹; 杨运文; 尚广东

    2013-01-01

    Recombineering is a genetic engineering technology for the DNA cloning and modification based on the recombinase-catalyzed homologous recombination. In this study, starting from previous obtained LS-GR, a chromosome-based recombineering strain,the 5'->3' single stranded DNA exonuclease coding gene recJ and exonucleolytic cleavage in the 3'->5' DNA exonuclease coding gene sbcB were knocked out via LS-GR's recombineering function as well as its inherent double strand break repair(DSBR) function. Compared with LS-GR, the obtained strains LS2002 and LS2004 show higher single strand oligonucleotide repair(SSOR) activity and DSBR activities, showing that they have the potential to be used in the recombineering experiments for high recombination efficiency.%重组工程是指通过重组酶催化DNA之间的同源重组,实现DNA克隆和修饰的一种基因工程技术.本研究从重组酶基因整合至大肠杆菌DH10B所获得的基于染色体的重组工程系统菌株LS-GR出发,利用菌株自身的重组工程系统和双链断裂修复功能,敲除了编码5'->3'单链DNA外切酶的recJ基因和编码3'->5'DNA外切核酸酶的sbcB基因,获得重组效率得以提高的菌株LS2002和LS2004.单链寡核苷酸修复和双链断裂修复实验表明所得菌株较LS-GR的重组效率有明显提高,对需要重组效率高的重组工程实验具有重要的应用价值.

  18. Enhanced and sustained CD8+ T cell responses with an adenoviral vector-based hepatitis C virus vaccine encoding NS3 linked to the MHC class II chaperone protein invariant chain

    DEFF Research Database (Denmark)

    Mikkelsen, Marianne; Holst, Peter Johannes; Bukh, Jens;

    2011-01-01

    Potent and broad cellular immune responses against the nonstructural (NS) proteins of hepatitis C virus (HCV) are associated with spontaneous viral clearance. In this study, we have improved the immunogenicity of an adenovirus (Ad)-based HCV vaccine by fusing NS3 from HCV (Strain J4; Genotype 1b...... memory. Functionally, the AdIiNS3-vaccinated mice had a significantly increased cytotoxic capacity compared with the AdNS3 group. The AdIiNS3-induced CD8(+) T cells protected mice from infection with recombinant vaccinia virus expressing HCV NS3 of heterologous 1b strains, and studies in knockout mice......) to the MHC class II chaperone protein invariant chain (Ii). We found that, after a single vaccination of C57BL/6 or BALB/c mice with Ad-IiNS3, the HCV NS3-specific CD8(+) T cell responses were significantly enhanced, accelerated, and prolonged compared with the vaccine encoding NS3 alone. The AdIiNS3...

  19. Development and evaluation of two truncated recombinant NP antigen-based indirect ELISAs for detection of bovine parainfluenza virus type 3 antibodies in cattle.

    Science.gov (United States)

    Yang, Yong; Wang, Feng-Xue; Sun, Na; Cao, Li; Zhang, Shu-Qin; Zhu, Hong-Wei; Guo, Li; Cheng, Shi-Peng; Wen, Yong-Jun

    2015-09-15

    Bovine parainfluenza virus type 3 (BPIV3) is one of the most important viral respiratory pathogens in both young and adult cattle. Nucleocapsid protein (NP) is the most abundant viral protein and the main regulator of virus replication and transcription. In this study, amino acid sequence data of BPIV3 NP was used to identify potential linear epitopic regions, which were subsequently used to design truncated recombinant NP antigens. The amino-terminal region (aa 9-157, NP-N) and the carboxy-terminal region (aa 391-500, NP-C) were selected, and these two truncated recombinant BPIV3 NP proteins were expressed in Escherichia coli based on the results of prediction studies. Furthermore, Enzyme-Linked Immunosorbent Assays (ELISAs) were established using the truncated recombinant BPIV3-N proteins as antigens, and 154 clinical samples were used to evaluate the newly established ELISA systems in comparison with a virus neutralisation test (VNT) as a reference. The results showed that a high coincidence rate was observed for the data that were obtained by the two methods. The sensitivity of NP-N ELISA and NP-C ELISA were 98.4% and 94.6%, respectively, and the specificity of both ELISAs was 100% with reference to the VNTs. Our data indicated that both ends of NP have high immunogenicity during BPIV3 infection and that they were good targets for serodiagnosis. The ELISAs based on the two truncated proteins were especially suitable for use in large-scale epidemiological investigations.

  20. Analysis of interchromosomal mitotic recombination.

    Science.gov (United States)

    McGill, C B; Shafer, B K; Higgins, D R; Strathern, J N

    1990-07-01

    A novel synthetic locus is described that provides a simple assay system for characterizing mitotic recombinants. The locus consists of the TRP1 and HIS3 genes inserted into chromosome III of S. cerevisiae between the CRY1 and MAT loci. Defined trp1 and his3 alleles have been generated that allow the selection of interchromosomal recombinants in this interval. Trp+ or His+ recombinants can be divided into several classes based on coupling of the other alleles in the interval. The tight linkage of the CRY1 and MAT loci, combined with the drug resistance and cell type phenotypes that they respectively control, facilitates the classification of the recombinants without resorting to tetrad dissection. We present the distribution of spontaneous recombinants among the classes defined by this analysis. The data suggest that the recombination intermediate can have regions of symmetric strand exchange and that co-conversion tracts can extend over 1-3 kb. Continuous conversion tracts are favored over discontinuous tracts. The distribution among the classes defined by this analysis is altered in recombinants induced by UV irradiation.

  1. Construction and characterization of calreticulin-HBsAg fusion gene recombinant adenovirus expression vector

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM: To generate recombinant adenoviral vector con-taining calreticulin (CRT)-hepatitis B surface antigen (HBsAg) fusion gene for developing a safe, effective and HBsAg-specific therapeutic vaccine.METHODS: CRT and HBsAg gene were fused using polymerase chain reaction (PCR), endonuclease diges-tion and ligation methods. The fusion gene was cloned into pENTR/D-TOPO transfer vector after the base pairs of DNA (CACC) sequence was added to the 5′ end. Adenoviral expression vector containing CRT-HBsAg fusion gen...

  2. Analysis of hepatitis B virus genotyping and drug resistance gene mutations based on massively parallel sequencing.

    Science.gov (United States)

    Han, Yingxin; Zhang, Yinxin; Mei, Yanhua; Wang, Yuqi; Liu, Tao; Guan, Yanfang; Tan, Deming; Liang, Yu; Yang, Ling; Yi, Xin

    2013-11-01

    Drug resistance to nucleoside analogs is a serious problem worldwide. Both drug resistance gene mutation detection and HBV genotyping are helpful for guiding clinical treatment. Total HBV DNA from 395 patients who were treated with single or multiple drugs including Lamivudine, Adefovir, Entecavir, Telbivudine, Tenofovir and Emtricitabine were sequenced using the HiSeq 2000 sequencing system and validated using the 3730 sequencing system. In addition, a mixed sample of HBV plasmid DNA was used to determine the cutoff value for HiSeq-sequencing, and 52 of the 395 samples were sequenced three times to evaluate the repeatability and stability of this technology. Of the 395 samples sequenced using both HiSeq and 3730 sequencing, the results from 346 were consistent, and the results from 49 were inconsistent. Among the 49 inconsistent results, 13 samples were detected as drug-resistance-positive using HiSeq but negative using 3730, and the other 36 samples showed a higher number of drug-resistance-positive gene mutations using HiSeq 2000 than using 3730. Gene mutations had an apparent frequency of 1% as assessed by the plasmid testing. Therefore, a 1% cutoff value was adopted. Furthermore, the experiment was repeated three times, and the same results were obtained in 49/52 samples using the HiSeq sequencing system. HiSeq sequencing can be used to analyze HBV gene mutations with high sensitivity, high fidelity, high throughput and automation and is a potential method for hepatitis B virus gene mutation detection and genotyping.

  3. Public health clinic-based hepatitis C testing and linkage to care in Baltimore.

    Science.gov (United States)

    Falade-Nwulia, O; Mehta, S H; Lasola, J; Latkin, C; Niculescu, A; O'Connor, C; Chaulk, P; Ghanem, K; Page, K R; Sulkowski, M S; Thomas, D L

    2016-05-01

    Testing and linkage to care are important determinants of hepatitis C virus (HCV) treatment effectiveness. Public health clinics serve populations at high risk of HCV. We investigated their potential to serve as sites for HCV testing, initiation of and linkage to HCV care. Cross-sectional study of patients accessing sexually transmitted infection (STI) care at the Baltimore City Health Department (BCHD) STI clinics, from June 2013 through April 2014 was conducted. Logistic regression was used to assess factors associated with HCV infection and specialist linkage to care. Between 24 June 2013 and 15 April 2014, 2681 patients were screened for HCV infection. Overall, 189 (7%) were anti-HCV positive, of whom 185 (98%) received follow-up HCV RNA testing, with 155 (84%) testing RNA positive. Of 155 RNA-positive individuals, 138 (89%) returned to the STI clinic for HCV RNA results and initial HCV care including counselling regarding transmission and harm reduction in alcohol, and 132 (85%) were referred to a specialist for HCV care. With provision of patient navigation services, 81 (52%) attended an offsite HCV specialist appointment. Alcohol use and lack of insurance coverage were associated with lower rates of specialist linkage (OR 0.4 [95% CI 0.1-0.9] and OR 0.4 [95% CI 0.1-0.9], respectively). We identified a high prevalence of HCV infection in BCHD STI clinics. With availability of patient navigation services, a large proportion of HCV-infected patients linked to off-site specialist care. PMID:26840570

  4. A new set of rDNA-NTS-based multiple integrative cassettes for the development of antibiotic-marker-free recombinant yeasts.

    Science.gov (United States)

    Moon, Hye Yun; Lee, Dong Wook; Sim, Gyu Hun; Kim, Hong-Jin; Hwang, Jee Youn; Kwon, Mun-Gyeong; Kang, Bo-Kyu; Kim, Jong Man; Kang, Hyun Ah

    2016-09-10

    The traditional yeast Saccharomyces cerevisiae has been widely used as a host system to produce recombinant proteins and metabolites of great commercial value. To engineer recombinant yeast that stably maintains expression cassettes without an antibiotic resistance gene, we developed new multiple integration cassettes by exploiting the non-transcribed spacer (NTS) of ribosomal DNA (rDNA) in combination with defective selection markers. The 5' and 3'-fragments of rDNA-NTS2 were used as flanking sequences for the expression cassettes carrying a set of URA3, LEU2, HIS3, and TRP1 selection markers with truncated promoters of different lengths. The integration numbers of NTS-based expression cassettes, ranging from one to ∼30 copies, showed a proportional increase with the extent of decreased expression of the auxotrophic markers. The NTS-based cassettes were used to construct yeast strains expressing the capsid protein of red-spotted grouper necrosis virus (RG-NNVCP) in a copy number-dependent manner. Oral administration of the recombinant yeast, harboring ∼30 copies of the integrated RG-NNVCP cassettes, provoked efficient immune responses in mice. In contrast, for the NTS cassettes expressing a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase, the integrant carrying only 4 copies was screened as the highest producer of squalene, showing a 150-fold increase compared to that of the wild-type strain. The multiple integrated cassettes were stably retained under prolonged nonselective conditions. Altogether, our results strongly support that rDNA-NTS integrative cassettes are useful tools to construct recombinant yeasts carrying optimal copies of a desired expression cassette without an antibiotic marker gene, which are suitable as oral vaccines or feed additives for animal and human consumption. PMID:27411901

  5. A new set of rDNA-NTS-based multiple integrative cassettes for the development of antibiotic-marker-free recombinant yeasts.

    Science.gov (United States)

    Moon, Hye Yun; Lee, Dong Wook; Sim, Gyu Hun; Kim, Hong-Jin; Hwang, Jee Youn; Kwon, Mun-Gyeong; Kang, Bo-Kyu; Kim, Jong Man; Kang, Hyun Ah

    2016-09-10

    The traditional yeast Saccharomyces cerevisiae has been widely used as a host system to produce recombinant proteins and metabolites of great commercial value. To engineer recombinant yeast that stably maintains expression cassettes without an antibiotic resistance gene, we developed new multiple integration cassettes by exploiting the non-transcribed spacer (NTS) of ribosomal DNA (rDNA) in combination with defective selection markers. The 5' and 3'-fragments of rDNA-NTS2 were used as flanking sequences for the expression cassettes carrying a set of URA3, LEU2, HIS3, and TRP1 selection markers with truncated promoters of different lengths. The integration numbers of NTS-based expression cassettes, ranging from one to ∼30 copies, showed a proportional increase with the extent of decreased expression of the auxotrophic markers. The NTS-based cassettes were used to construct yeast strains expressing the capsid protein of red-spotted grouper necrosis virus (RG-NNVCP) in a copy number-dependent manner. Oral administration of the recombinant yeast, harboring ∼30 copies of the integrated RG-NNVCP cassettes, provoked efficient immune responses in mice. In contrast, for the NTS cassettes expressing a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase, the integrant carrying only 4 copies was screened as the highest producer of squalene, showing a 150-fold increase compared to that of the wild-type strain. The multiple integrated cassettes were stably retained under prolonged nonselective conditions. Altogether, our results strongly support that rDNA-NTS integrative cassettes are useful tools to construct recombinant yeasts carrying optimal copies of a desired expression cassette without an antibiotic marker gene, which are suitable as oral vaccines or feed additives for animal and human consumption.

  6. Interferon-based therapy decreases risks of hepatocellular carcinoma and complications of cirrhosis in chronic hepatitis C patients.

    Directory of Open Access Journals (Sweden)

    Ching-Sheng Hsu

    Full Text Available BACKGROUND: Interferon-based therapy (IBT has been the standard of care for hepatitis C virus (HCV infection. However, conflicting results exist regarding the effects of IBT on risk of developing hepatocellular carcinoma (HCC and cirrhosis-associated complications, and most included highly selected patients. METHODS: This 8-year cohort study was based on the Longitudinal Health Insurance Database 2000 (LHID 2000 consisting of 1,000,000 beneficiaries randomly selected from all Taiwan National Health Insurance enrollees in 2000 (>23.7 million. Patients with newly detected HCV infections (n=11,264 were classified based on treatment and clinical outcomes. IBTs were defined as regimens that included interferon- alfa, pegylated interferon- alfa -2a, or pegylated interferon- alfa -2b for at least 3 months. The Cox proportional hazards models were used to estimate the hazard ratio (HR and associated confidence interval (CI of HCC and cirrhosis-associated complications for IBT. RESULTS: The 8-year incidence rate for HCC was 3.9% among patients who received IBT and 5.6% among those who did not. The HCC-free survival rate was significantly higher among patients receiving IBT during the 8-year period than their counterpart (adjusted HR, 0.50; 95% CI, 0.31-0.81; P= .004. Similarly, the event-free survival rates for esophageal variceal bleeding (adjusted HR, 0.45; 95% CI, 0.22-0.91; P= .026, hepatic encephalopathy (adjusted HR, 0.38; 95% CI, 0.21-0.69; P= .001, ascites (adjusted HR, 0.28; 95% CI, 0.14-0.57; P<.001, and cirrhosis (adjusted HR, 0.63; 95% CI, 0.44-0.91; P= .013 were significantly higher among patients who received IBT than those who did not, after adjustment for associated factors. CONCLUSION: Treatment with interferon may reduce the 8-year risk of HCC and cirrhosis-associated complications in patients with chronic HCV infection.

  7. PREVALENCE OF DIFFERENT VIRAL MARKERS IN PATIENTS OF ACUTE VIRAL HEPATITIS IN AND AROUND VISAKHAPATNAM : HOSPITAL BASED STUDY

    Directory of Open Access Journals (Sweden)

    Aruna Sree

    2015-04-01

    Full Text Available Acute viral hepatitis (AVH is a major public health problem and is an important cause of morbidity and mortality in the developing countries. AIM: The aim of the present study is to study the serological profile of acute viral hepatitis in children and adults admitted in King George Hospital, Visakhapatnam and also age and sex distribution of patients suffering from acute viral hepatitis and also comparing the etiological profile by studying serological markers of common viral agents. SUBJECTS AND METHODS: Samples were collected from 80 individuals with jaundice and other clinical and biochemical evidences of acute viral hepatitis . They were tested for hepatitis surface antigen, HBcIgM, HAVIgM, HEVIgM, Antibodies to HCV by the enzyme - linked immuno sorbent assay. RESULTS: Out of the 80 viral hepatitis cases (47 adults+33 children. In adults 20(42.5% patients presented HBV (26.96% was identified as the most common cause of acute hepatitis followed by HEV14 (29.8%, HEV+HAV4 (8.5% and HAV 6(12.76%. Co - infections with more than one virus were present in 5cases; HAV - HEV co - infection being the most common. In children 16(48.5% presented with HAV, HAV+HEV11 (33.3%, HEV4 (12.12%, HBV1 (3.03% CONCLUSIONS: Vaccination of adults against hepatitis B is indicated, along with sexual education to decrease the incidence of hepatitis which is found as common etiological agent in adults. The incidence of HAV and HEV in children shows that there is need for improvement in sanitation and food habits.

  8. A genetic validation study reveals a role of vitamin D metabolism in the response to interferon-alfa-based therapy of chronic hepatitis C.

    Directory of Open Access Journals (Sweden)

    Christian M Lange

    Full Text Available BACKGROUND: To perform a comprehensive study on the relationship between vitamin D metabolism and the response to interferon-α-based therapy of chronic hepatitis C. METHODOLOGY/PRINCIPAL FINDINGS: Associations between a functionally relevant polymorphism in the gene encoding the vitamin D 1α-hydroxylase (CYP27B1-1260 rs10877012 and the response to treatment with pegylated interferon-α (PEG-IFN-α and ribavirin were determined in 701 patients with chronic hepatitis C. In addition, associations between serum concentrations of 25-hydroxyvitamin D(3 (25[OH]D(3 and treatment outcome were analysed. CYP27B1-1260 rs10877012 was found to be an independent predictor of sustained virologic response (SVR in patients with poor-response IL28B genotypes (15% difference in SVR for rs10877012 genotype AA vs. CC, p = 0.02, OR = 1.52, 95% CI = 1.061-2.188, but not in patients with favourable IL28B genotype. Patients with chronic hepatitis C showed a high prevalence of vitamin D insufficiency (25[OH]D(3<20 ng/mL during all seasons, but 25(OHD(3 serum levels were not associated with treatment outcome. CONCLUSIONS/SIGNIFICANCE: Our study suggests a role of bioactive vitamin D (1,25[OH](2D(3, calcitriol in the response to treatment of chronic hepatitis C. However, serum concentration of the calcitriol precursor 25(OHD(3 is not a suitable predictor of treatment outcome.

  9. Genetic map of Triticum turgidum based on a hexaploid wheat population without genetic recombination for D genome

    Directory of Open Access Journals (Sweden)

    Zhang Li

    2012-08-01

    Full Text Available Abstract Background A synthetic doubled-haploid hexaploid wheat population, SynDH1, derived from the spontaneous chromosome doubling of triploid F1 hybrid plants obtained from the cross of hybrids Triticum turgidum ssp. durum line Langdon (LDN and ssp. turgidum line AS313, with Aegilops tauschii ssp. tauschii accession AS60, was previously constructed. SynDH1 is a tetraploidization-hexaploid doubled haploid (DH population because it contains recombinant A and B chromosomes from two different T. turgidum genotypes, while all the D chromosomes from Ae. tauschii are homogenous across the whole population. This paper reports the construction of a genetic map using this population. Results Of the 606 markers used to assemble the genetic map, 588 (97% were assigned to linkage groups. These included 513 Diversity Arrays Technology (DArT markers, 72 simple sequence repeat (SSR, one insertion site-based polymorphism (ISBP, and two high-molecular-weight glutenin subunit (HMW-GS markers. These markers were assigned to the 14 chromosomes, covering 2048.79 cM, with a mean distance of 3.48 cM between adjacent markers. This map showed good coverage of the A and B genome chromosomes, apart from 3A, 5A, 6A, and 4B. Compared with previously reported maps, most shared markers showed highly consistent orders. This map was successfully used to identify five quantitative trait loci (QTL, including two for spikelet number on chromosomes 7A and 5B, two for spike length on 7A and 3B, and one for 1000-grain weight on 4B. However, differences in crossability QTL between the two T. turgidum parents may explain the segregation distortion regions on chromosomes 1A, 3B, and 6B. Conclusions A genetic map of T. turgidum including 588 markers was constructed using a synthetic doubled haploid (SynDH hexaploid wheat population. Five QTLs for three agronomic traits were identified from this population. However, more markers are needed to increase the density and resolution of

  10. Evaluation on booster immunization efficacy of 5 μg hepatitis B vaccine made by recombinant deoxyribonucleic acid (DNA) techniques in polymorpha yeast of variant dosage in children aged over 5 years old%5岁以上儿童5μg重组乙型肝炎疫苗(酵母)加强免疫效果评价

    Institute of Scientific and Technical Information of China (English)

    陈永弟; 梁晓峰; 姚军; 崔富强; 王富珍; 沈灵智

    2012-01-01

    Objective To analyze the efficacy of booster immunization with domestic 5ug Hepatitis B Vaccine Made by Recombinant Deoxyribonucleic Acid (DNA) Techniques in Polymorpha Yeast ( HepB-Y) of variant dosage, in order to provide evidence for establishing immunization strategy. Methods 1728 children, with ages over 5 years were selected, who had been finished the basic immunization of hepatitis B vaccine in age under 1 year old. Blood plasma specimens of all sampled children were detected for hepatitis B virus ( HBV) surface antigen (HBsAg), antibodies to hepatitis B virus surface antigen (Anti-HBs) and antibodies to hepatitis B virus core antigen (Anti-HBc) by chemiluminescence. They were then classified into two groups of Anti-HBs positive and negative. Children of positive group were immunized one dosage of 5ug HepB-Y, while children of negative group were immunized three dosages of the same vaccine. Blood samples were collected after 1 month,and detected for Anti-HBs. Results The Anti-HBs positive rates were 40. 10%, 94. 04% and 99. 54% respectively of pre-immunization, post-immunization with one dosage and post-immunization with three dosages, there were statistical significant difference between any two among three rates (all P<0. 05). The Anti-HBs positive conversion rate of post-immunization with one dosage and three dosages were 88. 50% and 99. 42% respectively, the difference of positive conversion rate showed statistical significance between two groups (P< 0. 05). After immunization with one dosage in negative group, the aged rates of Anti-HBs positive conversion were dropping with age (P<0. 05). However, after immunization with three dosages, the aged rates of Anti-HBs positive conversion showed no relationship to age (P>0. 05). The average GMT of Anti-HBs negative children immunized with one dosage and three dosages were 450. 47mlu/ml and 664. 95mlu/ml respectively, while the average GMT of Anti-HBs positive children were 3663. 68mlu/ml after one

  11. Virus-like particle-based human vaccines: quality assessment based on structural and functional properties.

    Science.gov (United States)

    Zhao, Qinjian; Li, Shaowei; Yu, Hai; Xia, Ningshao; Modis, Yorgo

    2013-11-01

    Human vaccines against three viruses use recombinant virus-like particles (VLPs) as the antigen: hepatitis B virus, human papillomavirus, and hepatitis E virus. VLPs are excellent prophylactic vaccine antigens because they are self-assembling bionanoparticles (20 to 60 nm in diameter) that expose multiple epitopes on their surface and faithfully mimic the native virions. Here we summarize the long journey of these vaccines from bench to patients. The physical properties and structural features of each recombinant VLP vaccine are described. With the recent licensure of Hecolin against hepatitis E virus adding a third disease indication to prophylactic VLP-based vaccines, we review how the crucial quality attributes of VLP-based human vaccines against all three disease indications were assessed, controlled, and improved during bioprocessing through an array of structural and functional analyses.

  12. Recombinant Technology and Probiotics

    Directory of Open Access Journals (Sweden)

    Icy D’Silva

    2011-09-01

    Full Text Available Recombinant technology has led the way to monumental advances in the development of useful molecules, including the development of safe probiotics. The development of novel approaches using recombinant technology and probiotics that allow accurate targeting of therapeutics to the mucosa is an interesting area of research. The creation and use of recombinant probiotics expressing recombinantovalbumin, recombinant ovalbumin mutants and yet-to-be-designed recombinant hypo/non-allergenic molecules offer the opportunity to further investigate their effects for food, nutrition, environment andhealth. This review highlights advances in native probiotics and recombinant probiotics expressing native and recombinant molecules for food, nutrition, environment and health.

  13. Interface defects in SiC power MOSFETs - An electrically detected magnetic resonance study based on spin dependent recombination

    Energy Technology Data Exchange (ETDEWEB)

    Gruber, Gernot [KAI GmbH, Europastrasse 8, 9500 Villach, Austria and Graz University of Technology - Institute of Solid State Physics, Petersgasse 16, 8020 Graz (Austria); Hadley, Peter [Graz University of Technology - Institute of Solid State Physics, Petersgasse 16, 8020 Graz (Austria); Koch, Markus [Graz University of Technology - Institute of Experimental Physics, Petersgasse 16, 8020 Graz (Austria); Peters, Dethard [Infineon Technologies, Schottkystrasse 10, 91058 Erlangen (Germany); Aichinger, Thomas [Infineon Technologies, Siemensstrasse 2, 9500 Villach (Australia)

    2014-02-21

    This study presents electrically detected magnetic resonance (EDMR) measurements on a silicon carbide (SiC) MOSFET having the structure of a double-diffused silicon MOSFET (DMOS). The resonance pattern of a SiC DMOS was measured by monitoring the change of the recombination current between the source/body and the drain. The amplitude of the response has a maximum when the device is biased in depletion due to the equal concentrations of electrons and holes at the interface resulting in the most efficient recombination. The measured anisotropic g-tensor has axial symmetry with g{sub ∥} = 2.0051(4) (B ‖ c-axis), and g{sub ⊥} = 2.0029(4) (B⊥ c-axis) and the pattern shows several hyperfine (HF) peaks. We tentatively identify the observed defect as a silicon vacancy located directly at the interface.

  14. Bitargeted microemulsions based on coix seed ingredients for enhanced hepatic tumor delivery and synergistic therapy.

    Science.gov (United States)

    Qu, Ding; Sun, Wenjie; Liu, Mingjian; Liu, Yuping; Zhou, Jing; Chen, Yan

    2016-04-30

    A hepatic tumor bitargeted microemulsions drug delivery system using coix seed oil and coix seed polysaccharide (CP) acting as anticancer components, as well as functional excipients, was developed for enhanced tumor-specific accumulation by CP-mediated enhancement on passive tumor targeting and modification of galactose stearate (tumor-targeted ligand). In the physicochemical characteristics studies, galactose stearate-modified coix seed multicomponent microemulsions containing 30% CP (w%) (Gal-C-MEs) had a well-defined spherical shape with a small size (47.63 ± 1.41 nm), a narrow polydispersity index (PDI, 0.101 ± 0.002), and a nearly neutral surface charge (-4.37 ± 1.76 mV). The half-maximal inhibitory concentration (IC50) of Gal-C-MEs against HepG2 cells was 70.2 μg/mL, which decreased by 1.8-fold in comparison with that of coix seed multicomponent microemulsions (C-MEs). The fluorescence intensity of fluorescein isothiocyanate (FITC)-loaded Gal-C-MEs (FITC-Gal-C-MEs) internalized by HepG2 cells was 1.8-fold higher than that of FITC-loaded C-MEs (FIT C-C-MEs), but the cellular uptake of the latter became reduce by 1.6-fold when the weight ratio of CP decreased up to 10%. In the cell apoptosis studies, C-MEs (containing 30% CP) did not show a significant difference with Gal-C-MEs, but exhibited 3.3-fold and 1.5-fold increase relative to C-MEs containing 10% CP and 20% CP, respectively. In the in vivo tumor targeting studies, Cy5-loaded Gal-C-MEs (Cy5-Gal-C-MEs), notably distributed in the tumor sites and still found even at 48 h post-administration, displayed the strongest capability of tumor tissue accumulation and retention among all the test groups. Most importantly, Gal-C-MEs had stronger inhibition of tumor growth, prolonged survival time and more effectively tumor cell apoptosis induction in comparison with C-MEs containing different amounts of CP, which further confirmed that a certain amount of CP and tumor-targeted ligand were of great importance to

  15. Chronic Hepatitis C.

    Science.gov (United States)

    Tran, Tram T.; Martin, Paul

    2001-12-01

    Infection with hepatitis C virus (HCV) accounts for 40% of cases of chronic liver disease in the United States and is now the most common indication for liver transplantation. Estimates suggest that 4 million people (1.8%) of the American population are or have been infected with HCV. Currently, the treatment of choice for patients with chronic HCV infection is recombinant interferon alfa with ribavirin. Pegylated interferons are a promising new development, and in combination with ribavirin, they will rapidly become the standard of care. The goals of therapy are to slow disease progression, improve hepatic histology, reduce infectivity, and reduce the risk of hepatocellular carcinoma. Sustained virologic response, which generally implies the absence of viremia for 6 months or more following completion of therapy, is increasingly being regarded as a cure, with evidence of slowing or even regression of fibrosis on follow-up liver biopsy. A number of factors have been shown to be predictive of a sustained response, including viral genotype other than 1, low serum HCV RNA levels, absence of cirrhosis, younger age, female gender, and shorter duration of infection. Disease severity as assessed by liver biopsy, comorbidities, and possible contraindications to therapy should be weighed in the decision to begin treatment. Counseling patients regarding transmission, natural history, and drug and alcohol abstinence also should be included in management. Close monitoring should be done during treatment for side effects of interferon, including depression and bone marrow suppression. Hemolytic anemia is the major side effect of ribavirin. PMID:11696276

  16. Genetic map of Triticum turgidum based on a hexaploid wheat population without genetic recombination for D genome

    OpenAIRE

    Zhang Li; Luo Jiang-Tao; Hao Ming; Zhang Lian-Quan; Yuan Zhong-Wei; Yan Ze-Hong; Liu Ya-Xi; Zhang Bo; Liu Bao-Long; Liu Chun-Ji; Zhang Huai-Gang; Zheng You-Liang; Liu Deng-Cai

    2012-01-01

    Abstract Background A synthetic doubled-haploid hexaploid wheat population, SynDH1, derived from the spontaneous chromosome doubling of triploid F1 hybrid plants obtained from the cross of hybrids Triticum turgidum ssp. durum line Langdon (LDN) and ssp. turgidum line AS313, with Aegilops tauschii ssp. tauschii accession AS60, was previously constructed. SynDH1 is a tetraploidization-hexaploid doubled haploid (DH) population because it contains recombinant A and B chromosomes from two differen...

  17. Mechanism-Based Inhibition of Recombinant Human Cytochrome P450 3A4 by Tomato Juice Extract

    OpenAIRE

    須永, 克佳; 大川, 健一; 中村, 健一; 大久保, 温子; 原田, 園子; 津田, 整

    2012-01-01

    This study investigates whether tomato juice can inhibit cytochrome P450 (CYP) 3A4-mediated drug metabolism. Three commercially available, additive-free tomato juices, along with homogenized fresh tomato, were analyzed for their ability to inhibit testosterone 6β-hydroxylation activity using human recombinant CYP3A4. Results were compared to that of grapefruit juice. Ethyl acetate extracts of the tomato juices moderately reduced residual activity of CYP3A4 testosterone 6β-hydroxylation activi...

  18. Hepatic autoregulation

    DEFF Research Database (Denmark)

    Staehr, Peter; Hother-Nielsen, Ole; Beck-Nielsen, Henning;

    2007-01-01

    The effect of increased glycogenolysis, simulated by galactose's conversion to glucose, on the contribution of gluconeogenesis (GNG) to hepatic glucose production (GP) was determined. The conversion of galactose to glucose is by the same pathway as glycogen's conversion to glucose, i.e., glucose 1...

  19. Hepatic Encephalopathy

    Medline Plus

    Full Text Available ... of brain function in people with advanced liver disease. When your liver is damaged it can no longer remove toxic substances from your blood. These toxins build up and can travel through your body until they reach your brain, causing mental and physical symptoms of HE. Hepatic Encephalopathy often ...

  20. Hepatitis A vaccine associated with autoimmune hepatitis

    Institute of Scientific and Technical Information of China (English)

    PA Berry; G Smith-Laing

    2007-01-01

    To describe a case of probable relapsing autoimmune hepatitis associated with vaccination against hepatitis A virus (HAV). A case report and review of literature were written concerning autoimmune hepatitis in association with hepatitis A and other hepatotropic viruses. Soon after the administration of formalin-inactivated hepatitis A vaccine, a man who had recently recovered from an uncharacterized but self-limiting hepatitic illness,experienced a severe deterioration (AST 1687 U/L, INR 1.4). Anti-nuclear antibodies were detectable, and liver biopsy was compatible with autoimmune hepatitis. The observation supports the role of HAV as a trigger of autoimmune hepatitis. Studies in helper T-cell activity and antibody expression against hepatic proteins in the context of hepatitis A infection are summarized, and the concept of molecular mimicry with regard to other forms of viral hepatitis and autoimmunity is briefly explored.

  1. Direct costs of interferon-based and interferon-free direct-acting antiviral regimens for the treatment of chronic hepatitis C infection.

    Science.gov (United States)

    Gray, E; O'Leary, A; Kieran, J A; Fogarty, E; Dowling, T; Norris, S

    2016-09-01

    Given the increasing budget impact of Hepatitis C virus (HCV) treatment, robust real-world cost data are essential for healthcare decision-makers to evaluate and understand the costs and benefits of these treatments. To determine the direct cost of treating HCV infection in a hospital-based ambulatory care setting in Ireland based on available data from the Irish national hepatitis C treatment registry. A microcosting study of the direct costs of patients with hepatitis C treated with interferon-based and interferon-free direct-acting antiviral regimens was conducted. Attendance at the outpatient clinic for clinical assessment, the quantity of resources used per patient, the medication prescribed and the identification and timing of staff involvement was measured and combined to establish a mean cost of treatment per patient and a cost per sustained virological response (SVR). One hundred and sixty-eight patients were included in the analysis; 119 treated with interferon-based direct-acting antiviral regimens and 47 treated with interferon-free regimens. The mean costs of treatment with the interferon-based regimens per patient were €38 286 (95% CI €35 305-€41 061). The cost per SVR was €62 457. The mean cost of treatment with interferon-free regimens per patient was €55 734 (95% CI €50 906-€60 880). The cost per SVR was €81 873. Real-world cost data provide valuable information to enhance reimbursement decisions. While the direct costs associated with hepatitis C treatment in Ireland are substantial, it is reasonable to expect that the mean cost of treatment and the cost per SVR will reduce as patients with less advanced disease are treated with interferon-free therapies. PMID:26996144

  2. The combined effects of hospital and surgeon volume on short-term survival after hepatic resection in a population-based study.

    Directory of Open Access Journals (Sweden)

    Chun-Ming Chang

    Full Text Available BACKGROUND: The influence of different hospital and surgeon volumes on short-term survival after hepatic resection is not clearly clarified. By taking the known prognostic factors into account, the purpose of this study is to assess the combined effects of hospital and surgeon volume on short-term survival after hepatic resection. METHODS: 13,159 patients who underwent hepatic resection between 2002 and 2006 were identified in the Taiwan National Health Insurance Research Database. Data were extracted from it and short-term survivals were confirmed through 2006. The Cox proportional hazards model was used to assess the relationship between survival and different hospital, surgeon volume and caseload combinations. RESULTS: High-volume surgeons in high-volume hospitals had the highest short-term survivals, following by high-volume surgeons in low-volume hospitals, low-volume surgeons in high-volume hospitals and low-volume surgeons in low-volume hospitals. Based on Cox proportional hazard models, although high-volume hospitals and surgeons both showed significant lower risks of short-term mortality at hospital and surgeon level analysis, after combining hospital and surgeon volume into account, high-volume surgeons in high-volume hospitals had significantly better outcomes; the hazard ratio of other three caseload combinations ranging from 1.66 to 2.08 (p<0.001 in 3-month mortality, and 1.28 to 1.58 (p<0.01 in 1-year mortality. CONCLUSIONS: The combined effects of hospital and surgeon volume influenced the short-term survival after hepatic resection largely. After adjusting for the prognostic factors in the case mix, high-volume surgeons in high-volume hospitals had better short-term survivals. Centralization of hepatic resection to few surgeons and hospitals might improve patients' prognosis.

  3. Hepatitis A vaccine associated with autoimmune hepatitis

    OpenAIRE

    Berry, PA; Smith-Laing, G

    2007-01-01

    To describe a case of probable relapsing autoimmune hepatitis associated with vaccination against hepatitis A virus (HAV). A case report and review of literature were written concerning autoimmune hepatitis in association with hepatitis A and other hepatotropic viruses. Soon after the administration of formalin-inactivated hepatitis A vaccine, a man who had recently recovered from an uncharacterized but self-limiting hepatitic illness, experienced a severe deterioration (AST 1687 U/L, INR 1.4...

  4. Mechanism of Action and Antiviral Activity of Benzimidazole-Based Allosteric Inhibitors of the Hepatitis C Virus RNA-Dependent RNA Polymerase

    OpenAIRE

    Tomei, Licia; Altamura, Sergio; Bartholomew, Linda; Biroccio, Antonino; Ceccacci, Alessandra; Pacini, Laura; Narjes, Frank; Gennari, Nadia; Bisbocci, Monica; Incitti, Ilario; Orsatti, Laura; Harper, Steven; Stansfield, Ian; Rowley, Michael; De Francesco, Raffaele

    2003-01-01

    The RNA-dependent RNA polymerase of hepatitis C virus (HCV) is the catalytic subunit of the viral RNA amplification machinery and is an appealing target for the development of new therapeutic agents against HCV infection. Nonnucleoside inhibitors based on a benzimidazole scaffold have been recently reported. Compounds of this class are efficient inhibitors of HCV RNA replication in cell culture, thus providing attractive candidates for further development. Here we report the detailed analysis...

  5. Detection of Hepatitis C core antibody by dual-affinity yeast chimera and smartphone-based electrochemical sensing.

    Science.gov (United States)

    Aronoff-Spencer, Eliah; Venkatesh, A G; Sun, Alex; Brickner, Howard; Looney, David; Hall, Drew A

    2016-12-15

    Yeast cell lines were genetically engineered to display Hepatitis C virus (HCV) core antigen linked to gold binding peptide (GBP) as a dual-affinity biobrick chimera. These multifunctional yeast cells adhere to the gold sensor surface while simultaneously acting as a "renewable" capture reagent for anti-HCV core antibody. This streamlined functionalization and detection strategy removes the need for traditional purification and immobilization techniques. With this biobrick construct, both optical and electrochemical immunoassays were developed. The optical immunoassays demonstrated detection of anti-HCV core antibody down to 12.3pM concentrations while the electrochemical assay demonstrated higher binding constants and dynamic range. The electrochemical format and a custom, low-cost smartphone-based potentiostat ($20 USD) yielded comparable results to assays performed on a state-of-the-art electrochemical workstation. We propose this combination of synthetic biology and scalable, point-of-care sensing has potential to provide low-cost, cutting edge diagnostic capability for many pathogens in a variety of settings.

  6. Deletion modification enhances anthrax specific immunity and protective efficacy of a hepatitis B core particle-based anthrax epitope vaccine.

    Science.gov (United States)

    Yin, Ying; Zhang, Sheng; Cai, Chenguang; Zhang, Jun; Dong, Dayong; Guo, Qiang; Fu, Ling; Xu, Junjie; Chen, Wei

    2014-02-01

    Protective antigen (PA) is one of the major virulence factors of anthrax and is also the major constituent of the current anthrax vaccine. Previously, we found that the 2β2-2β3 loop of PA contains a dominant neutralizing epitope, the SFFD. We successfully inserted the 2β2-2β3 loop of PA into the major immunodominant region (MIR) of hepatitis B virus core (HBc) protein. The resulting fusion protein, termed HBc-N144-PA-loop2 (HBcL2), can effectively produce anthrax specific protective antibodies in an animal model. However, the protective immunity caused by HBcL2 could still be improved. In this research, we removed amino acids 79-81 from the HBc MIR of the HBcL2. This region was previously reported to be the major B cell epitope of HBc, and in keeping with this finding, we observed that the short deletion in the MIR not only diminished the intrinsic immunogenicity of HBc but also stimulated a higher titer of anthrax specific immunity. Most importantly, this deletion led to the full protection of the immunized mice against a lethal dose anthrax toxin challenge. We supposed that the conformational changes which occurred after the short deletion and foreign insertion in the MIR of HBc were the most likely reasons for the improvement in the immunogenicity of the HBc-based anthrax epitope vaccine.

  7. ED-based screening programs for hepatitis C (HCV) highlight significant opportunity to identify patients, prevent downstream costs/complications.

    Science.gov (United States)

    2014-01-01

    New data suggest there is a huge opportunity for EDs to identify patients with the hepatitis C virus (HCV) and link them into care before downstream complications lead to higher medical costs and adverse outcomes. Early results from a pilot study at the University of Alabama Medical Center in Birmingham show that at least 12% of the targeted baby boomer population being screened for HCV in the ED is testing positive for HCV, with confirmatory tests showing that about 9% of the screened population is infected with the disease. Both the Centers for Disease Control in Atlanta and the US Preventive Services Task Force recommend one-time HCV screening for patients who were born between 1945 and 1965. Public health experts say 75% of HCV infections occur in patients born during the baby boomer years, and that roughly half of them are unaware of their HCV status. Researchers at UAB report that so many patients are testing positive for HCV that demand for care can quickly overwhelm the health system if new primary care/specialty resources are not identified. Administrators of ED-based HCV screening programs in both Birmingham and Houston note that EDs with existing screening programs for HIV should have the easiest time implementing HCV screening. They also stress that patients are more accepting of HCV screening, and that the counseling process is easier. PMID:24432549

  8. Detection of Hepatitis C core antibody by dual-affinity yeast chimera and smartphone-based electrochemical sensing.

    Science.gov (United States)

    Aronoff-Spencer, Eliah; Venkatesh, A G; Sun, Alex; Brickner, Howard; Looney, David; Hall, Drew A

    2016-12-15

    Yeast cell lines were genetically engineered to display Hepatitis C virus (HCV) core antigen linked to gold binding peptide (GBP) as a dual-affinity biobrick chimera. These multifunctional yeast cells adhere to the gold sensor surface while simultaneously acting as a "renewable" capture reagent for anti-HCV core antibody. This streamlined functionalization and detection strategy removes the need for traditional purification and immobilization techniques. With this biobrick construct, both optical and electrochemical immunoassays were developed. The optical immunoassays demonstrated detection of anti-HCV core antibody down to 12.3pM concentrations while the electrochemical assay demonstrated higher binding constants and dynamic range. The electrochemical format and a custom, low-cost smartphone-based potentiostat ($20 USD) yielded comparable results to assays performed on a state-of-the-art electrochemical workstation. We propose this combination of synthetic biology and scalable, point-of-care sensing has potential to provide low-cost, cutting edge diagnostic capability for many pathogens in a variety of settings. PMID:27472403

  9. Using Molecular Initiating Events to Develop a Structural Alert Based Screening Workflow for Nuclear Receptor Ligands Associated with Hepatic Steatosis.

    Science.gov (United States)

    Mellor, Claire L; Steinmetz, Fabian P; Cronin, Mark T D

    2016-02-15

    In silico models are essential for the development of integrated alternative methods to identify organ level toxicity and lead toward the replacement of animal testing. These models include (quantitative) structure-activity relationships ((Q)SARs) and, importantly, the identification of structural alerts associated with defined toxicological end points. Structural alerts are able both to predict toxicity directly and assist in the formation of categories to facilitate read-across. They are particularly important to decipher the myriad mechanisms of action that result in organ level toxicity. The aim of this study was to develop novel structural alerts for nuclear receptor (NR) ligands that are associated with inducing hepatic steatosis and to show the vast number of existing data that are available. Current knowledge on NR agonists was extended with data from the ChEMBL database (12,713 chemicals in total) of bioactive molecules and from studying NR ligand-binding interactions within the protein database (PDB, 624 human NR structure files). A computational structural alert based workflow was developed using KNIME from these data using molecular fragments and other relevant chemical features. In total, 214 structural features were recorded computationally as SMARTS strings, and therefore, they can be used for grouping and screening during drug development and hazard assessment and provide knowledge to anchor adverse outcome pathways (AOPs) via their molecular initiating events (MIEs).

  10. Structure-Based Discovery of Novel Cyclophilin A Inhibitors for the Treatment of Hepatitis C Virus Infections.

    Science.gov (United States)

    Yang, Suhui; K R, Jyothi; Lim, Sangbin; Choi, Tae Gyu; Kim, Jin-Hwan; Akter, Salima; Jang, Miran; Ahn, Hyun-Jong; Kim, Hee-Young; Windisch, Marc P; Khadka, Daulat B; Zhao, Chao; Jin, Yifeng; Kang, Insug; Ha, Joohun; Oh, Byung-Chul; Kim, Meehyein; Kim, Sung Soo; Cho, Won-Jea

    2015-12-24

    Hepatitis C virus (HCV) is a major cause of end-stage liver disease. Direct-acting antivirals (DAAs), including inhibitors of nonstructural proteins (NS3/4A protease, NS5A, and NS5B polymerase), represent key components of anti-HCV treatment, but these are associated with increased drug resistance and toxicity. Thus, the development of host-targeted antiviral agents, such as cyclophilin A inhibitors, is an alternative approach for more effective, selective, and safer treatment. Starting with the discovery of a bis-amide derivative 5 through virtual screening, the lead compound 25 was developed using molecular modeling-based design and systematic exploration of the structure-activity relationship. The lead 25 lacked cytotoxicity, had potent anti-HCV activity, and showed selective and high binding affinity for CypA. Unlike cyclosporin A, 25 lacked immunosuppressive effects, successfully inhibited the HCV replication, restored host immune responses without acute toxicity in vitro and in vivo, and exhibited a high synergistic effect in combination with other drugs. These findings suggest that the bis-amides have significant potential to extend the arsenal of HCV therapeutics. PMID:26613291

  11. Deletion modification enhances anthrax specific immunity and protective efficacy of a hepatitis B core particle-based anthrax epitope vaccine.

    Science.gov (United States)

    Yin, Ying; Zhang, Sheng; Cai, Chenguang; Zhang, Jun; Dong, Dayong; Guo, Qiang; Fu, Ling; Xu, Junjie; Chen, Wei

    2014-02-01

    Protective antigen (PA) is one of the major virulence factors of anthrax and is also the major constituent of the current anthrax vaccine. Previously, we found that the 2β2-2β3 loop of PA contains a dominant neutralizing epitope, the SFFD. We successfully inserted the 2β2-2β3 loop of PA into the major immunodominant region (MIR) of hepatitis B virus core (HBc) protein. The resulting fusion protein, termed HBc-N144-PA-loop2 (HBcL2), can effectively produce anthrax specific protective antibodies in an animal model. However, the protective immunity caused by HBcL2 could still be improved. In this research, we removed amino acids 79-81 from the HBc MIR of the HBcL2. This region was previously reported to be the major B cell epitope of HBc, and in keeping with this finding, we observed that the short deletion in the MIR not only diminished the intrinsic immunogenicity of HBc but also stimulated a higher titer of anthrax specific immunity. Most importantly, this deletion led to the full protection of the immunized mice against a lethal dose anthrax toxin challenge. We supposed that the conformational changes which occurred after the short deletion and foreign insertion in the MIR of HBc were the most likely reasons for the improvement in the immunogenicity of the HBc-based anthrax epitope vaccine. PMID:24054942

  12. Research and Development of Hepatitis B Drugs: An Analysis Based on Technology Flows Measured by Patent Citations

    Science.gov (United States)

    Wan, Jian-bo; He, Chengwei; Hu, Yuanjia

    2016-01-01

    Despite the existence of available therapies, the Hepatitis B virus infection continues to be one of the most serious threats to human health, especially in developing countries such as China and India. To shed light on the improvement of current therapies and development of novel anti-HBV drugs, we thoroughly investigated 212 US patents of anti-HBV drugs and analyzed the technology flow in research and development of anti-HBV drugs based on data from IMS LifeCycle databases. Moreover, utilizing the patent citation method, which is an effective indicator of technology flow, we constructed patent citation network models and performed network analysis in order to reveal the features of different technology clusters. As a result, we identified the stagnant status of anti-HBV drug development and pointed the way for development of domestic pharmaceuticals in developing countries. We also discussed about therapeutic vaccines as the potential next generation therapy for HBV infection. Lastly, we depicted the cooperation between entities and found that novel forms of cooperation added diversity to the conventional form of cooperation within the pharmaceutical industry. In summary, our study provides inspiring insights for investors, policy makers, researchers, and other readers interested in anti-HBV drug development. PMID:27727319

  13. Risk factors associated with Hepatitis C among female substance users enrolled in community-based HIV prevention studies

    Directory of Open Access Journals (Sweden)

    O'Leary Catina C

    2011-04-01

    Full Text Available Abstract Background Hepatitis C virus (HCV infection is one of the most frequent chronic blood-borne infections in the United States. The epidemiology of HCV transmission is not completely understood, particularly in women and minorities. Findings We examined the HCV associated risk factors in substance abusing females involved in National Institute on Alcohol Abuse and Alcoholism (NIAAA and National Institute on Drug Abuse (NIDA funded HIV prevention studies of street recruited women. As a part of the 12 month follow-up, participants were interviewed about substance use and sexual risk behaviors, including drug implement sharing practices, tattoos, body piercing and blood transfusions and the sharing of personal hygiene equipment including tweezers, toothbrushes and razors. Urine and blood testing for HCV antibody (Ab, HIV and sexually transmitted diseases (STDs was conducted at the time of assessment. Among 782 predominantly African American women, 162 (21% tested positive for HCV Ab. Older age (p Conclusions This large community based sample of predominantly African American substance abusing women showed high prevalence of HCV Ab positivity and low awareness of their HCV serostatus. Our study demonstrated that in addition to intravenous drug use (IDU, other factors were significantly associated with HCV Ab positivity such as having a tattoo and a lifetime history of crack use. Other potential routes of HCV transmission should be further studied among high risk female populations.

  14. Hepatitis B virus taxonomy and hepatitis B virus genotypes

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Hepatitis B virus (HBV) is a member of the hepadnavirus family. Hepadnaviruses can be found in both mammals (orthohepadnaviruses) and birds (avihepadnaviruses).The genetic variability of HBV is very high. There are eight genotypes of HBV and three clades of HBV isolates from apes that appear to be additional genotypes of HBV. Most genotypes are now divided into subgenotypes with distinct virological and epidemiological properties. In addition, recombination among HBV genotypes increases the variability of HBV. This review summarises current knowledge of the epidemiology of genetic variability in hepadnaviruses and, due to rapid progress in the field,updates several recent reviews on HBV genotypes and subgenotypes.

  15. Psoriasis and hepatitis C virus.

    Science.gov (United States)

    Yamamoto, T; Katayama, I; Nishioka, K

    1995-11-01

    We have analyzed 8 patients (6 men and 2 women, aged 52 to 70 years) with psoriasis associated with hepatitis C virus (HCV) infection among 79 psoriatic patients. Psoriasis preceded in 6 cases. One patient had generalized pustular psoriasis (GPP), and the others had psoriasis vulgaris (PV). The psoriasis area and severity index (PASI) score ranged from 2.7 to 32.4. Two of the patients were treated with interferon-gamma. Anti-HCV antibodies were detected in all cases by second generation enzyme-linked immunosorbent and recombinant immunoblot assay. HCV messenger RNA was demonstrated by reverse transcriptase polymerase chain reaction in the tissue sections of the lesions of 1 of the patients with PV and the patient with GPP, providing evidence for active viral replication in the skin lesion. HCV-related chronic active hepatitis might cause several immunological abnormalities. It is suggested that this infection might be one of the triggering factors of psoriasis.

  16. Status of hepatitis B infection - a decade after hepatitis B vaccination of susceptible Nicobarese, an indigenous tribe of Andaman & Nicobar (A&N) islands with high hepatitis B endemicity

    OpenAIRE

    Bhattacharya, Haimanti; Bhattacharya, Debdutta; Ghosal, S.R.; Roy, Subarna; Sugunan, A. P.

    2015-01-01

    Background & objectives: Andaman and Nicobar Islands of India, home to six primitive tribes, constituting about 10 per cent of the total population of these Islands have been detected with high endemicity of hepatitis B infection. During 2000, a total of 936 individuals ≤ 45 yr, negative for hepatitis B surface antigen (HBsAg) and antibody anti-HBs were vaccinated with three doses of a recombinant DNA hepatitis B vaccine in two villages of Car Nicobar Islands. The present study was undertaken...

  17. Understanding of HLA-conferred susceptibility to chronic hepatitis B infection requires HLA genotyping-based association analysis

    Science.gov (United States)

    Nishida, Nao; Ohashi, Jun; Khor, Seik-Soon; Sugiyama, Masaya; Tsuchiura, Takayo; Sawai, Hiromi; Hino, Keisuke; Honda, Masao; Kaneko, Shuichi; Yatsuhashi, Hiroshi; Yokosuka, Osamu; Koike, Kazuhiko; Kurosaki, Masayuki; Izumi, Namiki; Korenaga, Masaaki; Kang, Jong-Hon; Tanaka, Eiji; Taketomi, Akinobu; Eguchi, Yuichiro; Sakamoto, Naoya; Yamamoto, Kazuhide; Tamori, Akihiro; Sakaida, Isao; Hige, Shuhei; Itoh, Yoshito; Mochida, Satoshi; Mita, Eiji; Takikawa, Yasuhiro; Ide, Tatsuya; Hiasa, Yoichi; Kojima, Hiroto; Yamamoto, Ken; Nakamura, Minoru; Saji, Hiroh; Sasazuki, Takehiko; Kanto, Tatsuya; Tokunaga, Katsushi; Mizokami, Masashi

    2016-01-01

    Associations of variants located in the HLA class II region with chronic hepatitis B (CHB) infection have been identified in Asian populations. Here, HLA imputation method was applied to determine HLA alleles using genome-wide SNP typing data of 1,975 Japanese individuals (1,033 HBV patients and 942 healthy controls). Together with data of an additional 1,481 Japanese healthy controls, association tests of six HLA loci including HLA-A, C, B, DRB1, DQB1, and DPB1, were performed. Although the strongest association was detected at a SNP located in the HLA-DP locus in a SNP-based GWAS using data from the 1,975 Japanese individuals, HLA genotyping-based analysis identified DQB1*06:01 as having the strongest association, showing a greater association with CHB susceptibility (OR = 1.76, P = 6.57 × 10−18) than any one of five HLA-DPB1 alleles that were previously reported as CHB susceptibility alleles. Moreover, HLA haplotype analysis showed that, among the five previously reported HLA-DPB1 susceptibility and protective alleles, the association of two DPB1 alleles (DPB1*09:01, and *04:01) had come from linkage disequilibrium with HLA-DR-DQ haplotypes, DRB1*15:02-DQB1*06:01 and DRB1*13:02-DQB1*06:04, respectively. The present study showed an example that SNP-based GWAS does not necessarily detect the primary susceptibility locus in the HLA region. PMID:27091392

  18. Recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibody to turkey coronavirus.

    Science.gov (United States)

    Abdelwahab, Mohamed; Loa, Chien Chang; Wu, Ching Ching; Lin, Tsang Long

    2015-06-01

    Nucleocapsid (N) protein gene of turkey coronavirus (TCoV) was expressed in a prokaryotic system and used to develop an enzyme-linked immunosorbent assay (ELISA) for detection of antibody to TCoV. Anti-TCoV hyperimmune turkey serum and normal turkey serum were used as positive or negative controls for optimization of the ELISA. Goat anti-turkey IgG (H+L) conjugated with horseradish peroxidase was used as detector antibody. Three hundred and twenty two turkey sera from the field were used to evaluate the performance of ELISA and determine the cut-off point of ELISA. The established ELISA was also examined with serum samples obtained from turkeys experimentally infected with TCoV. Those serum samples were collected at various time intervals from 1 to 63 days post-infection. The optimum conditions for differentiation between anti-TCoV hyperimmune serum and normal turkey serum were recombinant TCoV N protein concentration at 20 μg/ml, serum dilution at 1:800, and conjugate dilution at 1:10,000. Of the 322 sera from the field, 101 were positive for TCoV by immunofluorescent antibody assay (IFA). The sensitivity and specificity of the ELISA relative to IFA test were 86.0% and 96.8%, respectively, using the optimum cut-off point of 0.2 as determined by logistic regression method. Reactivity of anti-rotavirus, anti-reovirus, anti-adenovirus, or anti-enterovirus antibodies with the recombinant N protein coated on the ELISA plates was not detected. These results indicated that the established antibody-capture ELISA in conjunction with recombinant TCoV N protein as the coating protein can be utilized for detection of antibodies to TCoV in turkey flocks.

  19. d-lactate-selective amperometric biosensor based on the cell debris of the recombinant yeast Hansenula polymorpha.

    Science.gov (United States)

    Smutok, Oleh V; Dmytruk, Kostyantyn V; Karkovska, Maria I; Schuhmann, Wolfgang; Gonchar, Mykhailo V; Sibirny, Andriy A

    2014-07-01

    A d-lactate-selective biosensor has been developed using cells' debris of recombinant thermotolerant methylotrophic yeast Hansenula polymorpha, overproducing d-lactate: cytochrome c-oxidoreductase (EC 1.1.2.4, d-lactate dehydrogenase (cytochrome), DlDH). The H. polymorpha DlDH-producer was constructed in two steps. First, the gene CYB2 was deleted on the background of the С-105 (gcr1 catX) strain of H. polymorpha impaired in glucose repression and devoid of catalase activity to avoid specific l-lactate-cytochrome c oxidoreductase activity. Second, the homologous gene DLD1 coding for DlDH was overexpressed under the control of the strong H. polymorpha alcohol oxidase promoter in the frame of a plasmid for multicopy integration in the Δcyb2 strain. The selected recombinant strain possesses 6-fold increased DlDH activity as compared to the initial strain. The cells debris was used as a biorecognition element of a biosensor, since DlDH is strongly bound to mitochondrial membranes. The cells' debris, prepared by mechanic disintegration of recombinant cells, was immobilized on a graphite working electrode in an electrochemically generated layer using an Os-complex modified cathodic electrodeposition polymer. Cytochrome c was used as additional native electron mediator to improve electron transfer from reduced DlDH to the working electrode. The constructed d-lactate-selective biosensors are characterized by a high sensitivity (46.3-61.6 A M(-1)m(-2)), high selectivity and sufficient storage stability. PMID:24840438

  20. Evaluation of immune effect of recombinant fusion protein targeting the prostate stem cell antigen based on PSCA and HSP70

    Directory of Open Access Journals (Sweden)

    Lei DONG

    2014-10-01

    Full Text Available Objective To explore the immune effect and antitumor activity of recombinant prostate stem cell protein (PSCA and heat shock protein 70 (HSP70 in a murine model of prostate cancer. Methods Twenty-five healthy male C57BL/6 mice were randomly divided into 5 groups (5 each: PSCA, HSP, PSCA+HSP, PSCA-HSP and control group. Mice in the first 4 groups were vaccinated with the corresponding proteins, and those in control group were faked with injection of phosphate buffer saline (PBS. After immunization with recombinant proteins, the PSCA-specific cellular immune responses were monitored with ELISPOT, intracellular cytokine staining assay, and flow cytometry, and ELISA assay was used to detect humoral immune responses. The tumor growth and survival of vaccined mice were observed. Results ELISPOT revealed that the mice in PSCA-HSP group generated much more IFN-γ spot-forming cells than those in other groups (P<0.05, and they could generate strong anti-PSCA antibody response. Results of flow cytometry showed that the number of CD8+/IFN-γ+ T cells was significantly higher in PSCAHSP group than that in other groups (P<0.05. ELISA results revealed that all the mice in PSCA, PSCA+HSP and PSCA-HSP group were induced to generate the PSCA-specific humoral immune response, and no statistical difference was found on the antibody levels among the three groups. Animal experiment showed that PSCA-HSP could inhibit the growth of PSCA-expressing tumors and prolong the survival time of vaccinated mice. Conclusion HSP70 is a chaperone with significant effect for protein vaccines, and the recombinant fusion protein PSCA-HSP70 could be of potential value for prostate cancer treatment. DOI: 10.11855/j.issn.0577-7402.2014.09.08

  1. Hepatitis B immunization in adolescents.

    Science.gov (United States)

    Lawrence, M H; Goldstein, M A

    1995-10-01

    This article reviews the epidemiology of hepatitis B in the United States, previous vaccination strategy, and reasons for its failure and issues leading to the recommendation to vaccinate all adolescents. A review of specific hepatitis B virus risk behaviors of adolescents and barriers to vaccinating adolescents is covered. Strategies that favor successful completion of the immunization series are also examined. Hepatitis B infection is an important public health concern for adolescents. The previous vaccine strategy to immunize only individuals though to be at high risk was unsuccessful, especially because providers of care could not identify these individuals. Furthermore, many individuals thought not to be at high risk for infection were exposed through contacts which could not be identified. Challenges to immunization of adolescents include logistical issues, patient education, cost of the vaccine, and patient compliance. Several of these issues can be addressed by a school-based hepatitis B immunization program. The body of evidence and national policy is rapidly changing to support the recommendation that all adolescents receive the hepatitis B immunization series. The series would be most effective if administered during the middle-school years. A universal adolescent hepatitis B vaccination program would result in the most immediate health benefits and acceleration toward the eradication of hepatitis B in the United States. PMID:8580124

  2. Origins and Evolution of Hepatitis B Virus and Hepatitis D Virus.

    Science.gov (United States)

    Littlejohn, Margaret; Locarnini, Stephen; Yuen, Lilly

    2016-01-01

    Members of the family Hepadnaviridae fall into two subgroups: mammalian and avian. The detection of endogenous avian hepadnavirus DNA integrated into the genomes of zebra finches has revealed a deep evolutionary origin of hepadnaviruses that was not previously recognized, dating back at least 40 million and possibly >80 million years ago. The nonprimate mammalian members of the Hepadnaviridae include the woodchuck hepatitis virus (WHV), the ground squirrel hepatitis virus, and arctic squirrel hepatitis virus, as well as a number of members of the recently described bat hepatitis virus. The identification of hepatitis B viruses (HBVs) in higher primates, such as chimpanzee, gorilla, orangutan, and gibbons that cluster with the human HBV, as well as a number of recombinant forms between humans and primates, further implies a more complex origin of this virus. We discuss the current theories of the origin and evolution of HBV and propose a model that includes cross-species transmissions and subsequent recombination events on a genetic backbone of genotype C HBV infection. The hepatitis delta virus (HDV) is a defective RNA virus requiring the presence of the HBV for the completion of its life cycle. The origins of this virus remain unknown, although some recent studies have suggested an ancient African radiation. The age of the association between HDV and HBV is also unknown. PMID:26729756

  3. Electrical properties of III-Nitride LEDs: Recombination-based injection model and theoretical limits to electrical efficiency and electroluminescent cooling

    Science.gov (United States)

    David, Aurelien; Hurni, Christophe A.; Young, Nathan G.; Craven, Michael D.

    2016-08-01

    The current-voltage characteristic and ideality factor of III-Nitride quantum well light-emitting diodes (LEDs) grown on bulk GaN substrates are investigated. At operating temperature, these electrical properties exhibit a simple behavior. A model in which only active-region recombinations have a contribution to the LED current is found to account for experimental results. The limit of LED electrical efficiency is discussed based on the model and on thermodynamic arguments, and implications for electroluminescent cooling are examined.

  4. A novel, lactase-based selection and strain improvement strategy for recombinant protein expression in Kluyveromyces lactis

    Directory of Open Access Journals (Sweden)

    Krijger Jorrit-Jan

    2012-08-01

    Full Text Available Abstract Background The Crabtree-negative yeast species Kluyveromyces lactis has been established as an attractive microbial expression system for recombinant proteins at industrial scale. Its LAC genes allow for utilization of the inexpensive sugar lactose as a sole source of carbon and energy. Lactose efficiently induces the LAC4 promoter, which can be used to drive regulated expression of heterologous genes. So far, strain manipulation of K. lactis by homologous recombination was hampered by the high rate of non-homologous end-joining. Results Selection for growth on lactose was applied to target the insertion of heterologous genes downstream of the LAC4 promoter into the K. lactis genome and found to yield high numbers of positive transformants. Concurrent reconstitution of the β-galactosidase gene indicated the desired integration event of the expression cassette, and β-galactosidase activity measurements were used to monitor gene expression for strain improvement and fermentation optimization. The system was particularly improved by usage of a cell lysis resistant strain, VAK367-D4, which allowed for protein accumulation in long-term fermentation. Further optimization was achieved by increased gene dosage of KlGAL4 encoding the activator of lactose and galactose metabolic genes that led to elevated transcription rates. Pilot experiments were performed with strains expressing a single-chain antibody fragment (scFvox and a viral envelope protein (BVDV-E2, respectively. scFvox was shown to be secreted into the culture medium in an active, epitope-binding form indicating correct processing and protein folding; the E2 protein could be expressed intracellularly. Further data on the influence of protein toxicity on batch fermentation and potential post-transcriptional bottlenecks in protein accumulation were obtained. Conclusions A novel Kluyveromyces lactis host-vector system was developed that places heterologous genes under the control of

  5. Recombination with coat protein transgene in a complemen-tation system based on Cucumber mosaic virus (CMV)

    Institute of Scientific and Technical Information of China (English)

    LEI; Wanli

    2001-01-01

    plant cell suspension cultures, in Plant Tissue Culture Manual (ed. Lindsey, K.), Dordrecht: Kluwer Academic Publishers, 1991, A3: 1-21.[12]Damm, B., Willmitzer, L., Arabidopsis protoplast transformation and re-generation, in Plant Tissue Culture Manual (ed. Lindsey, K.), Dordrecht: Kluwer Academic Publishers, 1991, A7: 1-20.[13]Saunders, J. A., Rhodes, S. C., Kaper, J. M., Effects of electroporation pulse wave on the incorporation of viral RNA into tobacco protoplasts, BioTechniques, 1989, 7: 1124-1131.[14]Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature, 1970, 227: 680-685.[15]Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd ed., New York: Cold Spring Har-bor Laboratory Press, 1989, 18.60-18.75.[16]Ding, S. W., Rathjen, J. P., Li, W. X. et al., Efficient infection from cDNA clones of cucumber mosaic cucumovirus RNAs in a new plasmid vector, J. Gen. Virol., 1995, 76: 459-464.[17]Chomczynski, P., Sacchi, N., Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, Anal. Biochem., 1987, 162: 156-159.[18]Owen, J., Shintaku, M., Aeschleman, P., Nucleotide sequence and evolutionary relationships of cucumber mosaic virus (CMV) strains: CMV RNA3, J. Gen. Virol., 1990, 71: 2243-2249.[19]Roossinck, M. J., Zhang, L., Hellwald, K. H., Rearrangements in the 5′ nontranslated region and phylogenetic analysis of cucumber mosaic virus RNA3 indicate radial evolution of three subgroups, J. Virol., 1999, 73: 6752-6758.[20]Mori, M., Mise, K., Kobayashi, K., Infectivity of plasmids containing brome mosaic virus cDNA linked to the cauliflower mosaic virus 35S RNA promoter, J. Gen. Virol., 1991, 72: 243-246.[21]Aaziz, R., Tepfer, M., Recombination in RNA viruses and in virus-resistant transgenic plants, J. Gen. Virol., 1999, 80: 1339-1346.[22]Rubio, T., Borja, M., Scholthof, H. B. et al., Recombination with

  6. Interleukin-10 Enhances the Therapeutic Effectiveness of a Recombinant Poxvirus-Based Vaccine in an Experimental Murine Tumor Model

    OpenAIRE

    Kaufman, Howard L.; Rao, Jay B.; Irivine, Kari R.; Bronte, Vincenzo; Rosenberg, Steven A.; Restifo, Nicholas P

    1999-01-01

    Interleukin-10 (IL-10) has a wide range of in vivo biological activities and is a key regulatory cytokine of immune-mediated inflammation. The authors found that murine IL-10 given 12 hours after a recombinant vaccinia virus (rVV) containing the LacZ gene significantly enhanced the treatment of mice bearing 3-day-old pulmonary metastases expressing β-galactosidase. Because IL-10 has been shown to inhibit the functions of key elements of both innate and acquired immune responses, the authors h...

  7. Primordial magnetogenesis before recombination

    CERN Document Server

    Fabre, Ophélia

    2015-01-01

    The origin of large magnetic fields in the Universe remains currently unknown. We investigate here a mechanism before recombination based on known physics. The source of the vorticity is due to the changes in the photon distribution function caused by the fluctuations in the background photons. We show that the magnetic field generated in the MHD limit, due to the Coulomb scattering, is of the order $10^{-49}$ G. We explicitly show that the magnetic fields generated from this process are sustainable and are not erased by resistive diffusion. We compare the results with current observations and discuss the implications.

  8. DNA-based vaccination induces humoral and cellular immune responses against hepatitis B virus surface antigen in mice without activation of Cmyc

    Institute of Scientific and Technical Information of China (English)

    Lian San Zhao; Shan Qin; Tao You Zhou; Hong Tang; Li Liu; Bing Jun Lei

    2000-01-01

    AIM To develop a safe and effective DNA vaccine for inducing humoral and cellular immunological responses against hepatitis B virus surface antigen (HBsAg). METHODS BALB/c mice were inoculated with NV-HB/s, a recombinant plasmid that had been inserted S gene of hepatitis B virus genome and could express HBsAg in eukaryotes. HBsAg expression was measured by ABC immunohistochemical assay, generation of anti-HBs by ELISA and cytotoxic T lymphocyte (CTL), by MTT method, existence of vaccine DNA by Southern blot hybridization and activation of oncogene C-myc by in situ hybridization.RESULTS With NV-HB/s vaccination by intramuscular injection, anti-HBs was initially positive 2 weeks after inoculation while all mice tested were HBsAg positive in the muscles. The titers and seroconversion rate of anti-HBs were steadily increasing as time went on and were dose-dependent. All the mice inoculated with 100 μg NV-HB/ s were anti-HBs positive one month after inoculation, the titer was 1:1024 or more. The humoral immune response was similar induced by either intramuscular or intradermal injection. CTL activities were much stronger (45.26%) in NV-HB/s DNA immunized mice as compared with those (only 6%) in plasmaderived HBsAg vaccine immunized mice. Two months after inoculation, all muscle samples were positive by Southern-blot hybridization for NV-HB/s DNA detection, but decreased to 25%and all were undetectable by in situ hybridization after 6 months. No oncogene Cmyc activation was found in the muscle of inoculation site. CONCLUSION NV-HB/s could generate humoral and cellular immunological responses against HBsAg that had been safely expressed in situ by NV-HB/s vaccination.

  9. Protective T Cell and Antibody Immune Responses against Hepatitis C Virus Achieved Using a Biopolyester-Bead-Based Vaccine Delivery System.

    Science.gov (United States)

    Martínez-Donato, G; Piniella, B; Aguilar, D; Olivera, S; Pérez, A; Castañedo, Y; Alvarez-Lajonchere, L; Dueñas-Carrera, S; Lee, J W; Burr, N; Gonzalez-Miro, M; Rehm, B H A

    2016-04-01

    Hepatitis C virus (HCV) infection is a major worldwide problem. Chronic hepatitis C is recognized as one of the major causes of cirrhosis, hepatocellular carcinoma, and liver failure. Although new, directly acting antiviral therapies are suggested to overcome the low efficacy and adverse effects observed for the current standard of treatment, an effective vaccine would be the only way to certainly eradicate HCV infection. Recently, polyhydroxybutyrate beads produced by engineeredEscherichia colishowed efficacy as a vaccine delivery system. Here, an endotoxin-freeE. colistrain (ClearColi) was engineered to produce polyhydroxybutyrate beads displaying the core antigen on their surface (Beads-Core) and their immunogenicity was evaluated in BALB/c mice. Immunization with Beads-Core induced gamma interferon (IFN-γ) secretion and a functional T cell immune response against the HCV Core protein. With the aim to target broad T and B cell determinants described for HCV, Beads-Core mixed with HCV E1, E2, and NS3 recombinant proteins was also evaluated in BALB/c mice. Remarkably, only three immunization with Beads-Core+CoE1E2NS3/Alum (a mixture of 0.1 μg Co.120, 16.7 μg E1.340, 16.7 μg E2.680, and 10 μg NS3 adjuvanted in aluminum hydroxide [Alum]) induced a potent antibody response against E1 and E2 and a broad IFN-γ secretion and T cell response against Core and all coadministered antigens. This immunological response mediated protective immunity to viremia as assessed in a viral surrogate challenge model. Overall, it was shown that engineered biopolyester beads displaying foreign antigens are immunogenic and might present a particulate delivery system suitable for vaccination against HCV. PMID:26888185

  10. Microwave Ablation of Hepatic Malignancy

    OpenAIRE

    Lubner, Meghan G.; Brace, Christopher L.; Ziemlewicz, Tim J.; Hinshaw, J. Louis; Lee, Fred. T.

    2013-01-01

    Microwave ablation is an extremely promising heat-based thermal ablation modality that has particular applicability in treating hepatic malignancies. Microwaves can generate very high temperatures in very short time periods, potentially leading to improved treatment efficiency and larger ablation zones. As the available technology continues to improve, microwave ablation is emerging as a valuable alternative to radiofrequency ablation in the treatment of hepatic malignancies. This article rev...

  11. What Is Hepatitis?

    Science.gov (United States)

    ... Twitter Facebook Google + iTunes Play Store What is hepatitis? Online Q&A Reviewed July 2016 Q: What ... Question and answer archives Submit a question World Hepatitis Day Know hepatitis - Act now Event notice Key ...

  12. Living with Hepatitis

    Science.gov (United States)

    ... treatment of chronic hepatitis C (HCV) and hepatitis B (HBV) infections. Back to top Living With Hepatitis C In past years, many individuals learned that they have HCV from a blood test during a routine physical or because they attempted ...

  13. Hepatitis Risk Assessment

    Science.gov (United States)

    ... visit this page: About CDC.gov . Hepatitis Risk Assessment Recommend on Facebook Tweet Share Compartir Viral Hepatitis. ... at risk? Take this 5 minute Hepatitis Risk Assessment developed by the CDC and get a personalized ...

  14. Preventing hepatitis A

    Science.gov (United States)

    Hepatitis A is inflammation (irritation and swelling) of the liver caused by the hepatitis A virus. You can take several steps to ... reduce your risk of spreading or catching the hepatitis A virus: Always wash your hands thoroughly after ...

  15. Drug-induced hepatitis

    Science.gov (United States)

    Toxic hepatitis ... to get liver damage. Some drugs can cause hepatitis with small doses, even if the liver breakdown ... liver. Many different drugs can cause drug-induced hepatitis. Painkillers and fever reducers that contain acetaminophen are ...

  16. Hepatitis C FAQs

    Science.gov (United States)

    ... State and Local Partners & Grantees Resource Center Hepatitis C FAQs for the Public Recommend on Facebook Tweet ... URL - Redirecting ... Quick Links to Hepatitis ... A | B | C | D | E Viral Hepatitis Home Statistics & Surveillance Populations & ...

  17. Hepatitis B FAQs

    Science.gov (United States)

    ... State and Local Partners & Grantees Resource Center Hepatitis B FAQs for the Public Recommend on Facebook Tweet ... date URL - redirecting ... Quick Links to Hepatitis ... A | B | C | D | E Viral Hepatitis Home Statistics & Surveillance ...

  18. Hepatitis B Test

    Science.gov (United States)

    ... limited. Home Visit Global Sites Search Help? Hepatitis B Testing Share this page: Was this page helpful? Also known as: HBV Tests; Hep B; anti-HBs; Hepatitis B Surface Antibody; HBsAg; Hepatitis ...

  19. Hepatitis B virus (image)

    Science.gov (United States)

    Hepatitis B is also known as serum hepatitis and is spread through blood and sexual contact. It is ... population. This photograph is an electronmicroscopic image of hepatitis B virus particles. (Image courtesy of the Centers for ...

  20. Resistance-Associated NS5A Variants of Hepatitis C Virus Are Susceptible to Interferon-Based Therapy.

    Directory of Open Access Journals (Sweden)

    Jun Itakura

    Full Text Available The presence of resistance-associated variants (RAVs of hepatitis C virus (HCV attenuates the efficacy of direct acting antivirals (DAAs. The objective of this study was to characterize the susceptibility of RAVs to interferon-based therapy.Direct and deep sequencing were performed to detect Y93H RAV in the NS5A region. Twenty nine genotype 1b patients with detectable RAV at baseline were treated by a combination of simeprevir, pegylated interferon and ribavirin. The longitudinal changes in the proportion of Y93H RAV during therapy and at breakthrough or relapse were determined.By direct sequencing, Y93H RAV became undetectable or decreased in proportion at an early time point during therapy (within 7 days in 57% of patients with both the Y93H variant and wild type virus at baseline when HCV RNA was still detectable. By deep sequencing, the proportion of Y93H RAV against Y93 wild type was 52.7% (5.8%- 97.4% at baseline which significantly decreased to 29.7% (0.16%- 98.3% within 7 days of initiation of treatment (p = 0.023. The proportion of Y93H RAV was reduced in 21 of 29 cases (72.4% and a marked reduction of more than 10% was observed in 14 cases (48.7%. HCV RNA reduction was significantly greater for Y93H RAV (-3.65±1.3 logIU/mL/day than the Y93 wild type (-3.35±1.0 logIU/mL/day (p<0.001.Y93H RAV is more susceptible to interferon-based therapy than the Y93 wild type.