Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 30
Filter
1.
Bioorg Med Chem Lett ; 93: 129412, 2023 09 01.
Article in English | MEDLINE | ID: mdl-37499987

ABSTRACT

Small-molecule capsid assembly modulators (CAMs) have been recently recognized as promising antiviral agents for curing chronic hepatitis B virus (HBV) infection. A target-based in silico screening study is described, aimed towards the discovery of novel HBV CAMs. Initial optimization of four weakly active screening hits was performed via focused library synthesis. Lead compound 42 and close analogues 56 and 57 exhibited in vitro potency in the sub- and micromolar range along with good physico-chemical properties and were further evaluated in molecular docking and mechanism of action studies.


Subject(s)
Hepatitis B, Chronic , Hepatitis B , Humans , Hepatitis B virus , Capsid , Virus Assembly , Molecular Docking Simulation , Capsid Proteins , Antiviral Agents/pharmacology , Antiviral Agents/chemistry , Virus Replication
2.
Anal Methods ; 14(2): 135-146, 2022 01 06.
Article in English | MEDLINE | ID: mdl-34918017

ABSTRACT

To date, hepatitis B virus (HBV) capsid assembly modulators (CAMs), which target the viral core protein and induce the formation of non-functional viral capsids, have been identified and characterized in microtiter plate-based biochemical or cell-based in vitro assays. In this work, we developed an automated microfluidic screening assay, which uses convection-dominated Taylor-Aris dispersion to generate high-resolution dose-response curves, enabling the measurements of compound EC50 values at very short incubation times. The measurement of early kinetics down to 7.7 seconds in the microfluidic format was utilized to discriminate between the two different classes of CAMs known so far. The CAM (-N), leading to the formation of morphologically normal capsids and the CAM (-A), leading to aberrant HBV capsid structures. CAM-A compounds like BAY 41-4109 and GLS4 showed rapid kinetics, with assembly rates above 80% of the core protein after only a 7 second exposure to the compound, whereas CAM-N compounds like ABI-H0731 and JNJ-56136379 showed significantly slower kinetics. Using our microfluidic system, we characterized two of our in-house screening compounds. Interestingly, one compound showed a CAM-N/A intermediate behavior, which was verified with two standard methods for CAM classification, size exclusion chromatography, and anti-HBc immunofluorescence microscopy. With this proof-of-concept study, we believe that this microfluidic system is a robust primary screening tool for HBV CAM drug discovery, especially for the hit finding and hit-to-lead optimization phases. In addition to EC50 values, this system gives valuable first information about the mode of action of novel CAM screening compounds.


Subject(s)
Capsid , Hepatitis B virus , Antiviral Agents/chemistry , Antiviral Agents/metabolism , Antiviral Agents/pharmacology , Capsid/metabolism , Hepatitis B virus/metabolism , Microfluidics , Organic Chemicals
3.
Front Bioeng Biotechnol ; 9: 718889, 2021.
Article in English | MEDLINE | ID: mdl-34381768

ABSTRACT

Biosensors become increasingly relevant for medical diagnostics, pharmaceutical industry, and environmental technology, for example, to test new drugs easily and reliably or to detect cell growth in changing environmental conditions. Novel materials like graphene are promising candidates to produce biosensors on an industrial scale by means of printing processes. To reach this aim, methods for the reliable and automated production of electrode structures and their coating are required. We present an impedance biosensor in the format of a microtiter plate, fabricated by highly efficient roll-to-roll printing of graphene-based microstructures on large-area polymer foils. Proof-of-principle experiments show the evidence of the suitability of the printed graphene biosensors for impedance-based monitoring of viral cytopathogenicity and its inhibition in the presence of antiviral drugs. The developed system is a promising approach toward cost-efficient impedimetric biosensors for high-throughput screening in vaccine research and antiviral drug development.

4.
Arthritis Res Ther ; 22(1): 258, 2020 10 29.
Article in English | MEDLINE | ID: mdl-33121528

ABSTRACT

BACKGROUND: At present, etanercept represents the most commonly prescribed biologic agent for juvenile idiopathic arthritis (JIA) treatment. Children and adolescents with JIA are often treated with etanercept over long periods, sometimes even into adulthood. The objectives of this analysis were to determine the long-term safety of etanercept compared to a biologic-naïve cohort and to assess the long-term treatment response upon continuous etanercept exposure using data from the German biologics registry (BiKeR). METHODS: JIA patients newly exposed to etanercept were documented in the BiKeR registry from January 2001 to March 2019, and baseline characteristics, effectiveness, and safety parameters were analysed. Response to treatment was assessed according to 10-joint Juvenile Arthritis Disease Activity Score (JADAS10), JADAS-defined minimal disease activity and remission, JIA-American College of Rheumatology (ACR) improvement criteria, and ACR-inactive disease definition. Safety assessments were based on adverse event (AE) reports. RESULTS: A total of 2725 new etanercept users with a diagnosis of JIA were registered. Of these, etanercept was received as a first-line biologic by 95.8% and as monotherapy without concomitant methotrexate by 31.5%. After nine years on continuous treatment, 68.1% of patients presented minimal disease activity, 43.1% JADAS-defined remission on drug, and 36.6% ACR-inactive disease. JIA-ACR30/50/70/90 response rates were still 82/79/71/54% after nine years of treatment. Overall, 2053 AEs (34.3/100PY), including 226 serious AEs (SAE, 3.8/100PY), were observed upon etanercept, compared to 1345 AEs [35.6/100PY; p = 0.3] and 52 SAEs (1.4/100PY; p = 0.0001) in the biologic-naïve cohort. Respective exposure-adjusted rates for etanercept and biologic-naïve patients were 0.9/100PY and 0.2/100PY (p = 0.0001) for serious infections, 0.4/100PY and 0.1/100PY (p = 0.01) for zoster reactivation, 0.3/100PY and 0.03/100PY (p = 0.015) for inflammatory bowel disease, and 1.9/100PY and 1.4/100PY (p = 0.09) for uveitis. Three and two malignancies were documented in the etanercept and biologic-naïve groups, as well as three and one deaths, respectively. CONCLUSIONS: No new safety signal was observed, especially no increased risk for malignancies or autoimmune disorders other than inflammatory bowel disease. However, SAEs and serious infections, though infrequent, were more often reported on etanercept than in biologic-naïve patients. In addition, etanercept demonstrated a long-term maintenance of clinical benefits up to nine years of continuous treatment.


Subject(s)
Antirheumatic Agents , Arthritis, Juvenile , Adolescent , Adult , Antirheumatic Agents/adverse effects , Arthritis, Juvenile/drug therapy , Child , Etanercept/adverse effects , Humans , Registries , Treatment Outcome
5.
Rheumatology (Oxford) ; 59(9): 2287-2298, 2020 09 01.
Article in English | MEDLINE | ID: mdl-31846042

ABSTRACT

OBJECTIVE: Using data from the German Biologics JIA Registry (BIKER), long-term safety of biologics for systemic-onset JIA with regard to adverse events of special interest was assessed. METHODS: Safety assessments were based on adverse event reports after first dose through 90 days after last dose. Rates of adverse event, serious adverse event and 25 predefined adverse events of special interest were analysed. Incidence rates were compared for each biologic against all other biologics combined applying a mixed-effect Poisson model. RESULTS: Of 260 systemic-onset JIA patients in this analysis, 151 patients received etanercept, 109 tocilizumab, 71 anakinra and 51 canakinumab. Patients with etanercept had higher clinical Juvenile Arthritis Disease Activity Score 10 scores, active joint counts and steroid use at therapy start. Serious adverse events were reported with higher frequency in patients receiving canakinumab [20/100 patient years (PY)] and tocilizumab (21/100 PY). Cytopenia and hepatic events occurred with a higher frequency with tocilizumab and canakinumab. Medically important infections were seen more often in patients with IL-6 or IL-1 inhibition. Macrophage activation syndrome occurred in all cohorts with a higher frequency in patients with canakinumab (3.2/100 PY) and tocilizumab (2.5/100 PY) vs anakinra (0.83/100 PY) and etanercept (0.5/100 PY). After adjustment only an elevated risk for infections in anakinra-treated patients remained significant. Three definite malignancies were reported in patients ever exposed to biologics. Two deaths occurred in patients treated with etanercept. CONCLUSION: Surveillance of pharmacotherapy as provided by BIKER is an import approach especially for patients on long-term treatment. Overall, tolerance was acceptable. Differences between several biologics were noted and should be considered in daily patient care.


Subject(s)
Antibodies, Monoclonal, Humanized/adverse effects , Antirheumatic Agents/adverse effects , Arthritis, Juvenile/drug therapy , Biological Therapy/adverse effects , Drug-Related Side Effects and Adverse Reactions/epidemiology , Child, Preschool , Drug-Related Side Effects and Adverse Reactions/etiology , Etanercept/adverse effects , Female , Germany/epidemiology , Humans , Interleukin 1 Receptor Antagonist Protein/adverse effects , Macrophage Activation , Male , Product Surveillance, Postmarketing , Registries , Severity of Illness Index , Treatment Outcome
6.
Antiviral Res ; 158: 135-142, 2018 10.
Article in English | MEDLINE | ID: mdl-30031759

ABSTRACT

One of the most promising viral targets in current hepatitis B virus (HBV) drug development is the core protein due to its multiple roles in the viral life cycle. Here we investigated the differences in the mode of action and antiviral activity of representatives of six different capsid assembly modifier (CAM) scaffolds: three from the well-characterized scaffolds heteroarylpyrimidine (HAP), sulfamoylbenzamide (SBA), and phenylpropenamide (PPA), and three from novel scaffolds glyoxamide-pyrrolamide (GPA), pyrazolyl-thiazole (PT), and dibenzo-thiazepin-2-one (DBT). The target activity and antiviral efficacy of the different CAMs were tested in biochemical and cellular assays. Analytical size exclusion chromatography and transmission electron microscopy showed that only the HAP compound induced formation of aberrant non-capsid structures (class II mode of action), while the remaining CAMs did not affect capsid gross morphology (class I mode of action). Intracellular lysates from the HepAD38 cell line, inducibly replicating HBV, showed no reduction in the quantities of intracellular core protein or capsid after treatment with SBA, PPA, GPA, PT, or DBT compounds; however HAP-treatment led to a profound decrease in both. Additionally, immunofluorescence staining of compound-treated HepAD38 cells showed that all non-HAP CAMs led to a shift in the equilibrium of HBV core antigen (HBcAg) towards complete cytoplasmic staining, while the HAP induced accumulation of HBcAg aggregates in the nucleus. Our study demonstrates that the novel scaffolds GPA, PT, and DBT exhibit class I modes of action, alike SBA and PPA, whereas HAP remains the only scaffold belonging to class II inhibitors.


Subject(s)
Antiviral Agents/pharmacology , Capsid Proteins/drug effects , Capsid Proteins/metabolism , Capsid/drug effects , Hepatitis B virus/drug effects , Antiviral Agents/chemistry , Benzamides/chemistry , Benzamides/pharmacology , Benzoates , Cell Line , Drug Development , Hepatitis B Core Antigens , Hepatitis B virus/metabolism , Humans , Pyrimidines/chemistry , Pyrimidines/pharmacology , Viral Core Proteins , Virus Assembly/drug effects , Virus Replication/drug effects
7.
Pediatr Rheumatol Online J ; 16(1): 40, 2018 Jun 25.
Article in English | MEDLINE | ID: mdl-29940960

ABSTRACT

BACKGROUND: Juvenile dermatomyositis (JDM) is the most common inflammatory myopathy in childhood and a major cause of morbidity among children with pediatric rheumatic diseases. The management of JDM is very heterogeneous. The JDM working group of the Society for Pediatric Rheumatology (GKJR) aims to define consensus- and practice-based strategies in order to harmonize diagnosis, treatment and monitoring of JDM. METHODS: The JDM working group was established in 2015 consisting of 23 pediatric rheumatologists, pediatric neurologists and dermatologists with expertise in the management of JDM. Current practice patterns of management in JDM had previously been identified via an online survey among pediatric rheumatologists and neurologists. Using a consensus process consisting of online surveys and a face-to-face consensus conference statements were defined regarding the diagnosis, treatment and monitoring of JDM. During the conference consensus was achieved via nominal group technique. Voting took place using an electronic audience response system, and at least 80% consensus was required for individual statements. RESULTS: Overall 10 individual statements were developed, finally reaching a consensus of 92 to 100% regarding (1) establishing a diagnosis, (2) case definitions for the application of the strategies (moderate and severe JDM), (3) initial diagnostic testing, (4) monitoring and documentation, (5) treatment targets within the context of a treat-to-target strategy, (6) supportive therapies, (7) explicit definition of a treat-to-target strategy, (8) various glucocorticoid regimens, including intermittent intravenous methylprednisolone pulse and high-dose oral glucocorticoid therapies with tapering, (9) initial glucocorticoid-sparing therapy and (10) management of refractory disease. CONCLUSION: Using a consensus process among JDM experts, statements regarding the management of JDM were defined. These statements and the strategies aid in the management of patients with moderate and severe JDM.


Subject(s)
Dermatomyositis/drug therapy , Antirheumatic Agents/therapeutic use , Austria , Child , Consensus , Dermatomyositis/diagnosis , Disease Management , Germany , Glucocorticoids/therapeutic use , Humans , Surveys and Questionnaires
8.
Ann Thorac Surg ; 98(2): 723-5, 2014 Aug.
Article in English | MEDLINE | ID: mdl-25087804

ABSTRACT

The wide anatomic spectrum of Ebstein's anomaly is reflected by its extremely variable clinical presentation. We report a case of severe Ebstein's anomaly with natural history of a boy who underwent a successful emergency modified Starnes operation in a Third World sub-Saharan African country during a charitable international surgical mission. The particularities of this case are represented by the modality and setting of the intervention and by a surprising and fast clinical recovery. The age of the patient at the intervention and the choice to not fenestrate the patch also represent the noteworthiness of this life-saving procedure.


Subject(s)
Ebstein Anomaly/surgery , Emergency Treatment , Cardiac Surgical Procedures/methods , Child , Humans , Male , Remission Induction
9.
Thorac Cardiovasc Surg ; 62(5): 393-401, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24955755

ABSTRACT

BACKGROUND: Current data on cardiac surgery capacity on which to base effective concepts for developing sustainable cardiac surgical programs in Africa are lacking or of low quality. METHODS: A questionnaire concerning cardiac surgery in Africa was sent to 29 colleagues-26 cardiac surgeons and 3 cardiologists in 16 countries. Further, data on numbers of surgeons practicing in Africa were retrieved from the Cardiothoracic Surgery Network (CTSNet). RESULTS: There were 25 respondents, yielding a response rate of 86.2%. Three models emerged: the Ghanaian/German model with a senior local consultant surgeon (Model 1); surgeons visiting for a short period to perform humanitarian surgery (Model 2); and expatriate surgeons on contract to develop cardiac programs (Model 3). The 933 cardiothoracic surgeons listed by CTSNet translated into one surgeon per 1.3 million people. In North Africa, the figure was three surgeons per 1 million and in sub-Saharan Africa (SSA), one surgeon per 3.3 million people. The identified 156 cardiac surgeons represented a surgeon to population ratio of 1:5.9 million people. In SSA, the ratio was one surgeon per 14.3 million. In North Africa, it was one surgeon per 1.1 million people. Open heart operations were approximately 12 per million in Africa, 2 per million in SSA, and 92 per million people in North Africa. CONCLUSION: Cardiothoracic health care delivery would worsen in SSA without the support of humanitarian surgery. Although all three models have potential for success, the Ghanaian/German model has proved to be successful in the long term and could inspire health care policy makers and senior colleagues planning to establish cardiac programs in Africa.


Subject(s)
Cardiac Surgical Procedures/statistics & numerical data , Africa South of the Sahara/epidemiology , Africa, Northern/epidemiology , Cardiac Surgical Procedures/standards , Health Care Surveys , Health Policy , Humans , Program Development , Retrospective Studies
10.
Arthritis Care Res (Hoboken) ; 66(2): 253-62, 2014 Feb.
Article in English | MEDLINE | ID: mdl-23983081

ABSTRACT

OBJECTIVE: Improvement in health-related quality of life (HRQOL) is an important therapy goal in the treatment of patients with juvenile idiopathic arthritis (JIA). We investigated the 12-month course of HRQOL in patients with JIA after the start of therapy with etanercept and identified its determining factors. METHODS: Children with JIA were enrolled in the BiKer (Biologics in Pediatric Rheumatology) registry at the start of etanercept treatment. Children were prospectively followed in the first year of treatment and completed the Pediatric Quality of Life Inventory (PedsQL) at each occasion. The change in HRQOL was investigated by random-effect regression models. The time-varying variables pain and inactive disease were used for predicting the change in HRQOL. Inactive disease was defined by the Wallace et al criteria and pain was assessed on a visual analog scale (range 0-100). RESULTS: The children (n = 61) had a mean age of 10.6 years and a mean disease duration of 3.4 years at the start of etanercept. The mean PedsQL total score was 75. The PedsQL total score increased at a rate of 2.8 units per month (P < 0.001) in the first 6 months of treatment, up to a level of 89.7. A low HRQOL score was significantly highly associated with the number of tender joints, functional restrictions, pain, disease activity, and the existence of a comorbid condition at baseline. Inactive disease and reduced pain predicted better HRQOL under etanercept treatment. CONCLUSION: HRQOL was dramatically improved in children who started etanercept treatment. Inactive disease and lower pain were important predictors for improvement of HRQOL over time.


Subject(s)
Antirheumatic Agents/therapeutic use , Arthritis, Juvenile/drug therapy , Immunoglobulin G/therapeutic use , Quality of Life , Receptors, Tumor Necrosis Factor/therapeutic use , Adolescent , Arthritis, Juvenile/diagnosis , Arthritis, Juvenile/physiopathology , Arthritis, Juvenile/psychology , Child , Child, Preschool , Disability Evaluation , Etanercept , Female , Humans , Linear Models , Male , Multivariate Analysis , Pain Measurement , Prospective Studies , Registries , Severity of Illness Index , Surveys and Questionnaires , Time Factors , Treatment Outcome
11.
PLoS One ; 8(9): e74605, 2013.
Article in English | MEDLINE | ID: mdl-24066148

ABSTRACT

Inactivated orf virus (iORFV), strain D1701, is a potent immune modulator in various animal species. We recently demonstrated that iORFV induces strong antiviral activity in animal models of acute and chronic viral infections. In addition, we found D1701-mediated antifibrotic effects in different rat models of liver fibrosis. In the present study, we compare iORFV derived from two different strains of ORFV, D1701 and NZ2, respectively, with respect to their antifibrotic potential as well as their potential to induce an antiviral response controlling infections with the hepatotropic pathogens hepatitis C virus (HCV) and hepatitis B virus (HBV). Both strains of ORFV showed anti-viral activity against HCV in vitro and against HBV in a transgenic mouse model without signs of necro-inflammation in vivo. Our experiments suggest that the absence of liver damage is potentially mediated by iORFV-induced downregulation of antigen cross-presentation in liver sinus endothelial cells. Furthermore, both strains showed significant anti-fibrotic activity in rat models of liver fibrosis. iORFV strain NZ2 appeared more potent compared to strain D1701 with respect to both its antiviral and antifibrotic activity on the basis of dosages estimated by titration of active virus. These results show a potential therapeutic approach against two important human liver pathogens HBV and HCV that independently addresses concomitant liver fibrosis. Further studies are required to characterize the details of the mechanisms involved in this novel therapeutic principle.


Subject(s)
Hepacivirus/pathogenicity , Hepatitis B virus/pathogenicity , Liver Cirrhosis/virology , Orf virus/physiology , Animals , Humans , Liver Cirrhosis/prevention & control , Male , Mice , Rats , Swine
12.
Antiviral Res ; 95(2): 182-91, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22580131

ABSTRACT

The RNA-dependent RNA polymerase NS5B of the hepatitis C virus (HCV) has emerged as one of the key targets for antiviral drug discovery. Here we describe a novel non-nucleoside inhibitor (NNI) chemotype identified by screening: The substituted N-phenylbenzenesulphonamides (SPBS) which showed reversible inhibition of NS5B from HCV genotype 1b with IC(50) values up to 40 nM. Based on the decreased inhibitory activity against a recombinant NS5B protein carrying the mutation L419M or M423T we assumed that the SPBS inhibitors bind to the thumb site II which has already been described as the allosteric binding site for the NNI carboxy thiophene. The postulated binding site was consequently confirmed by solving two co-crystal structures of NS5B in complex with SPBS analogues at 2.3 and 2.2Å resolutions. The inhibitors are hydrogen-bonded to the main chain Ser476 and Tyr477 and to the side chain of Arg501. In addition, the inhibitors displayed van der Waals interactions with several residues of the hydrophobic binding pocket Leu419, Ile482, Leu497, Met423 and Trp528. Notably, the two SPBS analogues reported here revealed significant differences in addressing the NH-group of the main chain Tyr477 by hydrogen-bonds, water-mediated or directly, which provoked a shift of the carboxyphenyl group of the inhibitors towards the His475 position for the water-mediated binding mode. Interestingly, the differences observed in the binding mode led to a different cross resistance profile at positions M423 and I482. Using a panel of 38 individual NS5B proteins derived from different HCV genotypes, we could demonstrate inhibitory activity of the SPBS against polymerases from HCV genotypes 1a and 1b whereas the inhibitor class failed to inhibit any of the non-genotype 1 polymerases efficiently. Furthermore we demonstrated initial antiviral activity for SPBS against the subgenomic replicons of HCV genotypes 1a and 1b, respectively, and no considerable cytotoxic potential against a panel of ten different cell types.


Subject(s)
Antiviral Agents/pharmacology , Enzyme Inhibitors/pharmacology , Hepacivirus/drug effects , Hepacivirus/enzymology , Sulfonamides/pharmacology , Viral Nonstructural Proteins/antagonists & inhibitors , Binding Sites , Cell Line , Crystallography, X-Ray , Hepatocytes/virology , Humans , Inhibitory Concentration 50 , Microbial Sensitivity Tests , Models, Molecular , Protein Binding , Protein Conformation , Viral Nonstructural Proteins/chemistry , Viral Nonstructural Proteins/metabolism
13.
Antimicrob Agents Chemother ; 56(2): 1135-7, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22106211

ABSTRACT

AIC246 (letermovir) is a potent anticytomegalovirus drug in clinical development. Here, we report a consistent antiviral efficacy of AIC246 against human cytomegalovirus laboratory strains, clinical isolates, and virus variants resistant to approved drugs. Furthermore, we describe a remarkable selectivity of AIC246 for human cytomegaloviruses compared to that of other alpha-, beta-, or gammaherpesviruses or nonrelated pathogenic viruses, including adeno-, hepadna-, retro-, orthomyxo-, and flaviviruses. Our data confirm and support an excellent and selective anticytomegaloviral activity of AIC246.


Subject(s)
Antiviral Agents/pharmacology , Cytomegalovirus Infections/virology , Cytomegalovirus/drug effects , Quinazolines/pharmacology , Antiviral Agents/chemistry , Cytomegalovirus/isolation & purification , Drug Resistance, Viral/drug effects , Herpesviridae/classification , Herpesviridae/drug effects , Humans , Microbial Sensitivity Tests , Quinazolines/chemistry , Species Specificity , Virus Diseases/virology , Viruses/classification , Viruses/drug effects
14.
Biochim Biophys Acta ; 1814(10): 1325-32, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21621653

ABSTRACT

The RNA-dependent RNA polymerase of the hepatitis C virus (HCV) is the key enzyme for viral replication, recognized as one of the promising targets for antiviral intervention. Several of the known non-nucleoside HCV polymerase inhibitors (NNIs) identified by screening approaches show limitations in the coverage of all six major HCV genotypes (GTs). Genotypic profiling therefore has to be implemented early in the screening cascade to discover new broadly active NNIs. This implies knowledge of the specific individual biochemical properties of polymerases from all GTs which is to date limited to GT 1 only. This work gives a comprehensive overview of the biochemical properties of HCV polymerases derived from all major GTs 1-6. Biochemical analysis of polymerases from 38 individual sequences revealed that the optima for monovalent cations, pH and temperature were similar between the GTs, whereas significant differences concerning concentration of the preferred cofactor Mg(2+) were identified. Implementing the optimal requirements for the polymerases from each individual GT led to significant improvements in their enzymatic activities. However, the specific activity was distributed unequally across the GTs and could be ranked in the following descending order: 1b, 6a>2a, 3a, 4a, 5a>1a. Furthermore, the optimized assay conditions for genotypic profiling were confirmed by testing the inhibitory activity of 4 known prototype NNIs addressing the NNI binding sites 1 to 4.


Subject(s)
Antiviral Agents/pharmacology , Drug Resistance, Viral/genetics , Hepacivirus/enzymology , RNA-Dependent RNA Polymerase/genetics , RNA-Dependent RNA Polymerase/metabolism , Biochemical Phenomena/physiology , Cations, Monovalent/pharmacology , Drug Resistance, Viral/drug effects , Enzyme Activation/drug effects , Enzyme Activation/genetics , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Genotype , Hepacivirus/drug effects , Hepacivirus/genetics , Hepacivirus/metabolism , Hydrogen-Ion Concentration , Magnesium Chloride/pharmacology , Models, Biological , RNA-Dependent RNA Polymerase/antagonists & inhibitors , RNA-Dependent RNA Polymerase/chemistry , Structure-Activity Relationship , Temperature
15.
Circulation ; 122(12): 1159-66, 2010 Sep 21.
Article in English | MEDLINE | ID: mdl-20823387

ABSTRACT

BACKGROUND: Scimitar syndrome is a rare congenital heart disease. To evaluate the surgical results, we embarked on the European Congenital Heart Surgeons Association (ECHSA) multicentric study. METHODS AND RESULTS: From January 1997 to December 2007, we collected data on 68 patients who underwent surgery for scimitar syndrome. Primary outcomes included hospital mortality and the efficacy of repair at follow-up. Median age at surgery was 1.4 years (interquartile range, 0.46 to 7.92 years). Forty-four patients (64%) presented with symptoms. Surgical repair included intraatrial baffle in 38 patients (56%; group 1) and reimplantation of the scimitar vein onto the left atrium in 21 patients (31%; group 2). Eight patients underwent right pneumectomy, and 1 had a right lower lobe lobectomy (group 3). Four patients died in hospital (5.9%; 1 patient in group 1, 2.6%; 3 patients in group 3, 33%). Median follow-up time was 4.5 years. There were 2 late deaths (3.1%) resulting from severe pulmonary arterial hypertension. Freedom from scimitar drainage stenosis at 13 years was 83.8% in group 1 and 85.8% in group 2. Four patients in group 1 were reoperated, and 3 patients (2 in group 1 [6%] and 1 in group 2 [4.8%]) required balloon dilation/stenting for scimitar drainage stenosis. CONCLUSIONS: The surgical treatment of this rare syndrome is safe and effective. The majority of patients were asymptomatic at the follow-up control. There were a relatively high incidence of residual scimitar drainage stenosis that is similar between the 2 reported corrective surgical techniques used.


Subject(s)
Cardiovascular Surgical Procedures/methods , Scimitar Syndrome/surgery , Cardiovascular Surgical Procedures/adverse effects , Child , Child, Preschool , Constriction, Pathologic/epidemiology , Europe , Female , Follow-Up Studies , Hospital Mortality , Humans , Incidence , Infant , Kaplan-Meier Estimate , Male , Retrospective Studies , Scimitar Syndrome/mortality , Treatment Outcome
16.
Appl Environ Microbiol ; 73(20): 6436-43, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17720843

ABSTRACT

Cells containing reporters which are specifically induced via selected promoters are used in pharmaceutical drug discovery and in environmental biology. They are used in screening for novel drug candidates and in the detection of bioactive compounds in environmental samples. In this study, we generated and validated a set of five Bacillus subtilis promoters fused to the firefly luciferase reporter gene suitable for cell-based screening, enabling the as yet most-comprehensive high-throughput diagnosis of antibiotic interference in the major biosynthetic pathways of bacteria: the biosynthesis of DNA by the yorB promoter, of RNA by the yvgS promoter, of proteins by the yheI promoter, of the cell wall by the ypuA promoter, and of fatty acids by the fabHB promoter. The reporter cells mainly represent novel antibiotic biosensors compatible with high-throughput screening. We validated the strains by developing screens with a set of 14,000 pure natural products, representing a source of highly diverse chemical entities, many of them with antibiotic activity (6% with anti-Bacillus subtilis activity of

Subject(s)
Anti-Bacterial Agents/pharmacology , Bacillus subtilis/cytology , Bacillus subtilis/drug effects , Biosensing Techniques/methods , Bacillus subtilis/metabolism , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cell Membrane/drug effects , Cell Membrane/metabolism , DNA, Bacterial/antagonists & inhibitors , DNA, Bacterial/metabolism , Fatty Acids/antagonists & inhibitors , Fatty Acids/metabolism , Microbial Sensitivity Tests/methods , Promoter Regions, Genetic , RNA, Bacterial/antagonists & inhibitors , RNA, Bacterial/metabolism
17.
Eur J Cardiothorac Surg ; 31(5): 873-8, 2007 May.
Article in English | MEDLINE | ID: mdl-17339117

ABSTRACT

OBJECTIVE: Incidence of right ventricular outflow tract obstruction (RVOTO) may be suspected to be higher after arterial switch operation (ASO) for Taussig-Bing heart than after ASO for transposition of the great arteries (TGA), as Taussig-Bing anomaly is frequently associated with aortic arch obstruction and subvalvular aortic stenosis. We evaluated the risk to develop RVOTO after ASO for Taussig-Bing heart. METHODS: The 34 Taussig-Bing cases who underwent ASO from 1984 to 2005 were reviewed. RVOTO was defined as peak echo-gradient >or=30 mmHg across right ventricular outflow tract. Kaplan-Meier method was used to estimate time-related events. RESULTS: Subaortic stenosis was resected in 25 patients, 20 of whom (80%: 20/25) were discharged from hospital free from RVOTO. There was one early death: 2.9% mortality. Three patients died late. Actuarial survival was 85.1%+/-7.0% from 54 month onwards. Eleven survivors (36.7%: 11/30) experienced postoperative RVOTO. Obstruction was seen in 82% (9/11) of cases at subvalvular and/or valvular level. Surgery (n=4) or percutaneous intervention (n=2) was required in six patients. Patients discharged from hospital with RVOTO (n=8) were more likely to undergo reintervention for RVOTO (p=0.026). Freedom from reintervention for RVOTO decreased rapidly in the first two years to 86.5+/-6.3%, slowly thereafter (80.4+/-8.4% at year 7) and stabilized at 70.3+/-11.9% from year 11 on. Risk for RVOTO occurrence was 23.5+/-7.3% early after repair and progressively increased to level out at 53.6+/-11% at year 11. Patients who underwent subaortic resection were more likely (p=0.023) to be free from RVOTO occurrence or development. In the period under review, for patients who underwent ASO for simple (n=355) and complex (n=92) TGA, reoperation rate for neopulmonary stenosis was 0.3% (1/355) and 5.4% (5/92), respectively, to be compared to 11.8% (4/34) RVOTO rate of reoperation for Taussig-Bing heart in this study. CONCLUSIONS: Postoperative right-sided obstruction occurs more frequently after ASO repair of Taussig-Bing heart than after TGA arterial switching, leading to higher reintervention rate. Resection of the commonly associated subaortic stenosis often prevents RVOTO development.


Subject(s)
Double Outlet Right Ventricle/surgery , Postoperative Complications/etiology , Ventricular Outflow Obstruction/etiology , Aortic Stenosis, Subvalvular/complications , Aortic Stenosis, Subvalvular/mortality , Aortic Stenosis, Subvalvular/surgery , Cardiac Surgical Procedures/methods , Double Outlet Right Ventricle/complications , Double Outlet Right Ventricle/mortality , Echocardiography, Doppler , Heart Ventricles/physiopathology , Heart Ventricles/surgery , Humans , Infant , Infant, Newborn , Kaplan-Meier Estimate , Postoperative Complications/mortality , Postoperative Complications/physiopathology , Reoperation , Risk Assessment/methods , Risk Factors , Treatment Outcome , Ventricular Outflow Obstruction/mortality , Ventricular Outflow Obstruction/physiopathology
18.
Ann Thorac Surg ; 83(1): 161-8, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17184653

ABSTRACT

BACKGROUND: Surgery for congenital heart disease (CHD) has changed considerably during the last three decades. The results of primary repair have steadily improved, to allow treating almost all patients within the pediatric age; nonetheless an increasing population of adult patients requires surgical treatment. The objective of this study is to present the early surgical results of patients who require surgery for CHD in the adult population within a multicentered European study population. METHODS: Data relative to the hospital course of 2,012 adult patients (age > or = 18 years) who required surgical treatment for CHD from January 1, 1997 through December 31, 2004 were reviewed. Nineteen cardiothoracic centers from 13 European countries contributed to the data collection. RESULTS: Mean age at surgery was 34.4 +/- 14.53 years. Most of the operations were corrective procedures (1,509 patients, 75%), followed by reoperations (464 patients, 23.1%) and palliative procedures (39 patients, 1.9%). Six hundred forty-nine patients (32.2%) required surgical closure of an isolated ostium secundum atrial septal defect. Overall hospital mortality was 2%. Preoperative cyanosis, arrhythmias, and NYHA class III-IV, proved significant risk factors for hospital mortality. Follow-up data were available in 1,342 of 1,972 patients (68%) who were discharged home. Late deaths occurred in 6 patients (0.5%). Overall survival probability was 97% at 60 months, which is higher for corrective procedures (98.2%) if compared with reoperations (94.1%) and palliations (86.1%). CONCLUSIONS: Surgical treatment of CHD in adult patients, in specialized cardiac units, proved quite safe, beneficial, and low-risk.


Subject(s)
Cardiac Surgical Procedures/adverse effects , Heart Defects, Congenital/surgery , Adult , Cardiac Surgical Procedures/mortality , Female , Hospital Mortality , Humans , Male , Middle Aged , Postoperative Complications/epidemiology , Risk
19.
J Thorac Cardiovasc Surg ; 132(3): 633-9, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16935120

ABSTRACT

OBJECTIVES: This study analyzes the results of the arterial switch operation for transposition of the great arteries in member institutions of the European Congenital Heart Surgeons Association. METHODS: The records of 613 patients who underwent primary arterial switch operations in each of 19 participating institutions in the period from January 1998 through December 2000 were reviewed retrospectively. RESULTS: A ventricular septal defect was present in 186 (30%) patients. Coronary anatomy was type A in 69% of the patients, and aortic arch pathology was present in 20% of patients with ventricular septal defect. Rashkind septostomy was performed in 75% of the patients, and 69% received prostaglandin. There were 37 hospital deaths (operative mortality, 6%), 13 (3%) for patients with an intact ventricular septum and 24 (13%) for those with a ventricular septal defect (P < .001). In 36% delayed sternal closure was performed, 8% required peritoneal dialysis, and 2% required mechanical circulatory support. Median ventilation time was 58 hours, and intensive care and hospital stay were 6 and 14 days, respectively. Although of various preoperative risk factors the presence of a ventricular septal defect, arch pathology, and coronary anomalies were univariate predictors of operative mortality, only the presence of a ventricular septal defect approached statistical significance (P = .06) on multivariable analysis. Of various operative parameters, aortic crossclamp time and delayed sternal closure were also univariate predictors; however, only the latter was an independent statistically significant predictor of death. CONCLUSIONS: Results of the procedure in European centers are compatible with those in the literature. The presence of a ventricular septal defect is the clinically most important preoperative risk factor for operative death, approaching statistical significance on multivariable analysis.


Subject(s)
Transposition of Great Vessels/surgery , Europe , Female , Humans , Infant , Infant, Newborn , Male , Multivariate Analysis , Retrospective Studies , Vascular Surgical Procedures/methods
20.
J Thorac Cardiovasc Surg ; 132(1): 50-7, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16798302

ABSTRACT

OBJECTIVE: Since most centers' experience with Ebstein anomaly is limited, we sought to analyze the collective experience of participating institutions of the European Congenital Heart Surgeons Association with surgery for this rare malformation. METHODS: The records of all 150 patients (median age 6.4 years) who underwent surgery for Ebstein anomaly in the 13 participating Association centers between January 1992 and January 2005 were reviewed retrospectively. Patients with congenitally corrected transposition were excluded. RESULTS: Most patients (81%) had Ebstein disease type B or C and significant functional impairment (61% in New York Heart Association class III or IV) and 16% had prior operations. Surgical procedures (n = 179) included valve replacement (n = 60, 33.5%), valve repair (n = 49, 27.3%), 1(1/2) ventricle repair (n = 46, 25.6%), palliative shunt (n = 13, 7.26%), and other complex procedures (n = 11, 6.14%). There were 20 hospital deaths (operative mortality 13.3%) after valve replacement in 5 patients, valve repair in 3, 1(1/2) ventricle repair in 7, palliative procedures in 3, and miscellaneous procedures in 2. Younger age and palliative procedures were univariate risk factors for operative death, but only age was an independent predictor on multivariable analysis. CONCLUSIONS: Most patients coming to surgery presented in childhood and were significantly symptomatic. More than half underwent valve replacement or repair, but a considerable proportion had severe disease necessitating 1(1/2) ventricle repair or palliative procedures. Operative mortality did not differ significantly among repair, replacement, and 1(1/2) ventricle repair but was associated with palliative procedures for severe disease early in life, young age being the only independent predictor of operative death.


Subject(s)
Ebstein Anomaly/surgery , Adolescent , Adult , Age Distribution , Child , Child, Preschool , Ebstein Anomaly/mortality , Europe , Female , Heart Valves/surgery , Heart Ventricles/surgery , Hospital Mortality , Humans , Infant , Infant, Newborn , Logistic Models , Male , Middle Aged , Multivariate Analysis , Palliative Care , Retrospective Studies , Treatment Outcome
SELECTION OF CITATIONS
SEARCH DETAIL
...