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1.
Urologe A ; 54(5): 696-702, 2015 May.
Article in German | MEDLINE | ID: mdl-25616765

ABSTRACT

BACKGROUND: Patient counseling by using the IIEF to assess erectile function (EF) before and after radical prostatectomy (RPX) is only possible under limited circumstances. The aim of this study was to evaluate if the Erection Hardness Score (EHS) could be used in addition to the IIEF for the assessment of EF and patient preference regarding counseling for their sex life. MATERIAL AND METHODS: EF was evaluated in 307 patients 3-60 months after RPX using the IIEF-EF and EHS. Questionnaires assessed sexual activity/intercourse as well as satisfaction with sex life irrespective of EF (10-point Likert scale). Patients were further asked concerning development of new sexual methods independent of erection firm enough for penetration and further wishes regarding counseling for their sex life. RESULTS: Of 272 patients, 82.0% underwent bilateral nerve-sparing prostatectomy, 30.5% (n=83; mean age: 68.1 years) had sexual intercourse and 41.9% (n=114) were sexually active. EH Scores 1-2 and 4 coincided with compatible IIEF-EF Scores 1-21, and ≥ 26, respectively. Of the patients with an EHS of 3, 55.9% had an IIEF-EF score that was notably lower. Of patients with sexual intercourse, 65.8% were satisfied with their sex life; 53.2% of sexually active patients were satisfied without sexual intercourse. Alternative methods were manual/oral stimulation, cuddling, and the use of vibrators. Patients request individually tailored, realistic counseling. CONCLUSION: The advantage of the EHS compared to the IIEF is that the erectile function can be assessed irrespective of sexual intercourse and sexual partner. Counseling should assist patients towards the attainment of a satisfying sex life-even without an erection.


Subject(s)
Diagnostic Techniques, Urological , Erectile Dysfunction/diagnosis , Erectile Dysfunction/psychology , Patient Satisfaction/statistics & numerical data , Prostatectomy/psychology , Sex Counseling/statistics & numerical data , Aged , Aged, 80 and over , Erectile Dysfunction/etiology , Germany , Humans , Male , Middle Aged , Prostatectomy/adverse effects , Reproducibility of Results , Sensitivity and Specificity , Severity of Illness Index , Sexual Behavior/psychology , Sexual Behavior/statistics & numerical data , Sexuality , Treatment Outcome
2.
Int J Med Microbiol ; 305(3): 289-97, 2015 May.
Article in English | MEDLINE | ID: mdl-25592264

ABSTRACT

Streptococcus (S.) pneumoniae is a major cause of secondary bacterial pneumonia during influenza epidemics. Neuraminidase (NA) is a virulence factor of both pneumococci and influenza viruses. Bacterial neuraminidases (NAs) are structurally related to viral NA and susceptible to oseltamivir, an inhibitor designed to target viral NA. This prompted us to evaluate the antipneumococcal potential of two NA inhibiting natural compounds, the diarylheptanoid katsumadain A and the isoprenylated flavone artocarpin. Chemiluminescence, fluorescence-, and hemagglutination-based enzyme assays were applied to determine the inhibitory efficiency (IC(50) value) of the tested compounds towards pneumococcal NAs. The mechanism of inhibition was studied via enzyme kinetics with recombinant NanA NA. Unlike oseltamivir, which competes with the natural substrate of NA, artocarpin exhibits a mixed-type inhibition with a Ki value of 9.70 µM. Remarkably, artocarpin was the only NA inhibitor (NAI) for which an inhibitory effect on pneumococcal growth (MIC: 0.99-5.75 µM) and biofilm formation (MBIC: 1.15-2.97 µM) was observable. In addition, we discovered that the bactericidal effect of artocarpin can reduce the viability of pneumococci by a factor of >1000, without obvious harm to lung epithelial cells. This renders artocarpin a promising natural product for further investigations.


Subject(s)
Anti-Bacterial Agents/pharmacology , Enzyme Inhibitors/pharmacology , Mannose-Binding Lectins/pharmacology , Neuraminidase/antagonists & inhibitors , Plant Lectins/pharmacology , Streptococcus pneumoniae/drug effects , Biofilms/drug effects , Cell Line , Cell Survival/drug effects , Diarylheptanoids/pharmacology , Epithelial Cells/drug effects , Epithelial Cells/physiology , Humans , Inhibitory Concentration 50 , Kinetics , Mannose-Binding Lectins/toxicity , Microbial Sensitivity Tests , Microbial Viability/drug effects , Neuraminidase/metabolism , Plant Lectins/toxicity , Streptococcus pneumoniae/growth & development , Streptococcus pneumoniae/physiology
3.
J Med Virol ; 79(9): 1334-42, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17607777

ABSTRACT

The reasons for the different outcome of coxsackievirus B3 (CVB3)-induced heart disease in humans are not well understood. Since there are no experimental data on the course of disease after infection with genetically different CVB3 in a natural variable population until now, we studied the outcome of virus infection in outbred NMRI mice after inoculation of genetically different CVB3 variants. Adult male mice were inoculated with seven closely related CVB3 variants. The histopathological changes of heart and pancreas tissue, antibody induction, virus titers, and persistence of viral positive- as well as negative-strand RNA in spleen and heart tissue were compared at day 7 or day 28 after infection to detect prerequisites and predictive factors for chronic myocarditis. Six CVB3 variants infected NMRI mice. CVB3 infection (i) did not induce detectable myocardial injury, (ii) caused signs of healing up acute myocarditis or (iii) ongoing chronic myocarditis. Neither IgG antibody responses nor the extent of destruction of exocrine pancreatic tissue or viral RNA load in spleen did correlate with myocardial histopathology. In contrast, a high persistent viral RNA load in heart tissue specimens was characteristic for mice developing chronic myocarditis. The results of the present study corroborate high viral load in the acute stage of myocarditis and high amounts of persisting CVB3 RNA in heart tissue as predictive marker of chronic myocarditis. The outcome of CVB3-induced heart disease in outbred NMRI mice depends strongly on the viral genetic background. In particular an important role of viral capsid proteins is suggested.


Subject(s)
Coxsackievirus Infections/virology , Enterovirus B, Human/genetics , Myocarditis/virology , Animals , Antibodies, Viral/blood , Coxsackievirus Infections/immunology , Disease Models, Animal , Enterovirus B, Human/immunology , Enterovirus B, Human/physiology , Heart/virology , Male , Mice , Myocarditis/immunology , Myocardium/pathology , Pancreas/pathology , RNA, Viral/isolation & purification , Spleen/virology , Viral Load
4.
J Antimicrob Chemother ; 60(1): 68-77, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17550890

ABSTRACT

OBJECTIVES: The objectives of this study were (i) to apply computer-based technologies to evaluate the structure of 48 N,N'-(bis-5-nitropyrimidyl)dispirotripiperazines which belong to a new class of highly active antiviral compounds binding to cell surface heparan sulphates, (ii) to understand the chemical- biological interactions governing their activities, and (iii) to design new compounds with strong antiviral activity. METHODS: The logarithm of 50% cytotoxic concentration (CC(50)) in GMK cells, of 50% inhibitory concentration (IC(50)) against herpes simplex virus type 1, and of selectivity index (SI = CC(50)/IC(50)) was used to develop quantitative structure-activity relationships (QSARs) based on simplex representation of molecular structure. The QSAR model was applied to design new compounds. Two of these compounds were synthesized, physico-chemically characterized and tested for cytotoxicity and antiviral activity. RESULTS: Statistic characteristics for partial least squares models allow the prediction of CC(50), IC(50) and SI values. The QSAR results demonstrate a high impact of individual structural fragments for antiviral activity. Molecular fragments that promote and interfere with antiviral activity were defined on the basis of the obtained models. Electrostatic factors (38%) and hydrophobicity (34%) were the most important determinants of antiherpetic activity. Using the established method, new potential dispirotripiperazine derivatives were computationally designed. Two of these computationally designed compounds were synthesized. The biological test results confirm the computationally predicted values of these compounds. CONCLUSIONS: The established QSAR model is suitable for the design of new antiherpetic compounds and prediction of their activity.


Subject(s)
Antiviral Agents , Herpesvirus 1, Human/drug effects , Piperazines , Spiro Compounds , Antiviral Agents/chemistry , Antiviral Agents/metabolism , Antiviral Agents/pharmacology , Antiviral Agents/toxicity , Cell Line , Computational Biology , Cytopathogenic Effect, Viral , Drug Design , Heparitin Sulfate/metabolism , Humans , Magnetic Resonance Spectroscopy , Models, Molecular , Piperazines/chemistry , Piperazines/metabolism , Piperazines/pharmacology , Piperazines/toxicity , Spectrometry, Mass, Fast Atom Bombardment , Spectrophotometry, Infrared , Spiro Compounds/chemistry , Spiro Compounds/metabolism , Spiro Compounds/pharmacology , Spiro Compounds/toxicity , Structure-Activity Relationship
5.
Med Microbiol Immunol ; 195(2): 65-71, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16059699

ABSTRACT

Previous studies have shown that embryonated egg provides a convenient and easy to use system for in vivo screening of anti-influenza virus inhibitors. However, it is not known whether this model is suitable for testing neuraminidase (NA) inhibitors, too. Therefore, the present study describes the evaluation of the ion-channel blockers amantadine and rimantadine in comparison with the NA inhibitors oseltamivir and zanamivir by using the influenza A virus hen's egg model. The treatment was started immediately before or after the challenge dose was placed on the chorioallantoic membrane (CAM). Differences between the survival rate of treated and untreated chick embryos infected with influenza A virus were analyzed statistically. As result, the survival rate of chick embryos could be significantly increased when the treatment with amantadine, rimantadine, oseltamivir, or zanamivir was started before the CAM was inoculated with one egg infective dose 50% (EID50) influenza A virus. When the drugs were administered shortly after viral inoculation, significant antiviral efficacy was shown for rimantadine, oseltamivir, and zanamivir. Antiviral efficacy could be demonstrated exclusively for both oseltamivir and zanamivir after the embryos were infected with higher challenge doses of 10(2) EID50 influenza A virus. In conclusion, the NA inhibitors oseltamivir and zanamivir have a significantly better antiviral activity against influenza A virus than amantadine and rimantadine tested in embryonated hen's eggs. Therefore, this model can be a valuable alternative approach for in vivo pre-testing anti-influenza virus activity of NA inhibitors.


Subject(s)
Antiviral Agents/pharmacology , Enzyme Inhibitors/pharmacology , Influenza A virus/drug effects , Microbial Sensitivity Tests/methods , Neuraminidase/antagonists & inhibitors , Acetamides/pharmacology , Amantadine/pharmacology , Animals , Chick Embryo , Chickens , Guanidines/pharmacology , Influenza A virus/enzymology , Influenza in Birds/drug therapy , Oseltamivir , Pyrans/pharmacology , Rimantadine/pharmacology , Sialic Acids/pharmacology , Zanamivir
6.
J Ethnopharmacol ; 104(1-2): 182-7, 2006 Mar 08.
Article in English | MEDLINE | ID: mdl-16233967

ABSTRACT

Acokanthera schimperi (Apocynaceae), Euclea schimperi (Ebenaceae), Inula confertiflora (Asteraceae), Melilotus elegans (Leguminosae), and Plumbago zeylanica (Plumbaginaceae), are some of the medicinal plants used in Ethiopia for treatment of various skin disorders. In this study, the antiviral activities of the 80% methanolic extracts of these plants have been examined against coxsackievirus B3 (CVB3), influenza A virus and herpes simplex virus type1 Kupka (HSV-1) using cytopathic effect (CPE) inhibitory assays in HeLa, MDCK, and GMK cells, respectively. In parallel, the cytotoxicity was quantified using a crystal violet uptake assay. The antiviral activity of the most active compound was confirmed with plaque reduction assays. The results revealed that the extracts of Acokanthera schimperi and Euclea schimperi showed antiviral activity against all three tested viruses albeit with unequal efficacy. Whereas the Acokanthera schimperi extract exhibited the strongest activity against CVB3, the extract of Euclea schimperi inhibited influenzavirus A replication most effectively. A weak anti-influenzavirus A activity was also exhibited by the other plant extracts tested. In addition, CVB3 was inhibited by the extracts of Plumbago zeylanica and HSV-1 by Inula confertiflora. Thus, the extracts of these plants, particularly those of Acokanthera schimperi, Euclea schimperi and Inula confertiflora which showed activity against CVB3 and HSV-1 support their traditional use in the treatment of skin diseases of viral origin.


Subject(s)
Antiviral Agents/therapeutic use , Plants, Medicinal , Skin Diseases/drug therapy , Skin Diseases/virology , Animals , Antiviral Agents/isolation & purification , Chlorocebus aethiops , Dogs , Dose-Response Relationship, Drug , Ethiopia , HeLa Cells , Humans , Plant Extracts/isolation & purification , Plant Extracts/therapeutic use , Plant Structures
7.
J Virol ; 77(18): 10071-7, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12941917

ABSTRACT

Amino acid exchanges in the virus capsid protein VP1 allow the coxsackievirus B3 variant PD (CVB3 PD) to replicate in decay accelerating factor (DAF)-negative and coxsackievirus-adenovirus receptor (CAR)-negative cells. This suggests that molecules other than DAF and CAR are involved in attachment of this CVB3 variant to cell surfaces. The observation that productive infection associated with cytopathic effect occurred in Chinese hamster ovary (CHO-K1) cells, whereas heparinase-treated CHO-K1 cells, glucosaminoglycan-negative pgsA-745, heparan sulfate (HS)-negative pgsD-677, and pgsE-606 cells with significantly reduced N-sulfate expression resist CVB3 PD infection, indicates a critical role of highly sulfated HS. 2-O-sulfate-lacking pgsF-17 cells represented the cell line with minimum HS modifications susceptible for CVB3 PD. Inhibition of virus replication in CHO-K1 cells by polycationic compounds, pentosan polysulfate, lung heparin, and several intestinal but not kidney HS supported the hypothesis that CVB3 PD uses specific modified HS for entry. In addition, recombinant human hepatocyte growth factor blocked CVB3 PD infection. However, CAR also mediates CVB3 PD infection, because this CVB3 variant replicates in HS-lacking but CAR-bearing Raji cells, infection could be prevented by pretreatment of cells with CAR antibody, and HS-negative pgsD-677 cells transfected with CAR became susceptible for CVB3 PD. These results demonstrate that the amino acid substitutions in the viral capsid protein VP1 enable CVB3 PD to use specific modified HS as an entry receptor in addition to CAR.


Subject(s)
Capsid Proteins/physiology , Coxsackievirus Infections/etiology , Enterovirus B, Human/physiology , Receptors, Virus/physiology , Amino Acid Substitution , Animals , CHO Cells , Capsid Proteins/chemistry , Coxsackie and Adenovirus Receptor-Like Membrane Protein , Cricetinae , Heparin/pharmacology , Heparin Lyase/pharmacology , Heparitin Sulfate/physiology , Hepatocyte Growth Factor/pharmacology , Transfection
8.
Virology ; 311(1): 134-43, 2003 Jun 20.
Article in English | MEDLINE | ID: mdl-12832211

ABSTRACT

N,N'-bisheteryl derivatives of dispirotripiperazine (DSTP) are a novel class of antiviral compounds with some of their representatives very effectively inhibiting the replication of herpes simplex virus type 1 (HSV-1) in cell culture. Using one representative of these compounds, the N,N'-bis(1-oxido[1,2,5]oxadiazolo[3,4-d]pyrimidin-7-yl)-3,12-diaza-6,9-diazonia(5,2,5,2)dispirohexadecane dichloride (DSTP 27), we here further tried to elucidate the molecular mechanisms responsible for the antiviral activity. The results from plaque reduction assays under a variety of conditions suggest that inhibition of HSV-1 strain Kupka replication by DSTP 27 occurs at the level of viral attachment by blockade of heparan sulfate (HS) structures on the cell surface that are used as viral receptors. In contrast to heparin and pentosan polysulfate, pretreatment of cells with DSTP 27 resulted in efficient inhibition of viral adsorption and replication persisting several hours after removal of the inhibitor. Specific binding of DSTP 27 to heparin was demonstrated in vitro. Titrations of gC-positive and gC-negative pseudorabies virus (PrV) mutants on HS-positive and HS-negative cell lines confirmed that inhibitory action of DSTP 27 is strictly HS dependent. Aside from HSV-1 Kupka and PrV, DSTP 27 efficiently inhibits growth of several HSV-1 and HSV-2 strains, among them aciclovir/foscarnet-resistant strains, human cytomegalovirus, human respiratory syncytial virus, and human immunodeficiency viruses known to attach to the cell surface via HS.


Subject(s)
Antiviral Agents/pharmacology , Heparitin Sulfate/metabolism , Herpesvirus 1, Human/drug effects , Herpesvirus 1, Suid/drug effects , Oxadiazoles/pharmacology , Piperazines/pharmacology , Pyrimidines/pharmacology , Receptors, Virus/drug effects , Animals , Cells, Cultured , Dose-Response Relationship, Drug , Heparitin Sulfate/chemistry , Herpesvirus 1, Human/pathogenicity , Herpesvirus 1, Suid/genetics , Herpesvirus 2, Human/drug effects , Humans , Mutation , Oxadiazoles/metabolism , Pyrimidines/metabolism , Receptors, Virus/metabolism , Virus Replication/drug effects
9.
Fitoterapia ; 74(1-2): 139-43, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12628410

ABSTRACT

The crude extracts of the leaves of Dodonaea viscosa and Rumex nervosus as well as of the root of Rumex abyssinicus were tested for anti-microbial and anti-inflammatory activities. It was observed that the three plants possess antibacterial activity against Streptococcus pyogenes and Staphylococcus aureus and strong activity against Coxsackie virus B3 and influenza A virus. In contrast, none of them exhibited anti-fungal activity. The anti-inflammatory activity test results verified that only R. abyssinicus inhibited the synthesis of prostaglandin (PG) E(2).


Subject(s)
Anti-Infective Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Phytotherapy , Plant Extracts/pharmacology , Rumex , Sapindaceae , Animals , Anti-Infective Agents/administration & dosage , Anti-Infective Agents/therapeutic use , Anti-Inflammatory Agents/administration & dosage , Anti-Inflammatory Agents/therapeutic use , Cell Line, Tumor/drug effects , Dose-Response Relationship, Drug , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , HeLa Cells/drug effects , Humans , Macrophages/drug effects , Mice , Mitosporic Fungi/drug effects , Plant Extracts/administration & dosage , Plant Extracts/therapeutic use , Plant Leaves , Plant Roots
11.
Antiviral Res ; 55(1): 117-27, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12076756

ABSTRACT

During the search for new antivirals, various N,N'-bis-5-pyrimidyl derivatives of 3,12-diaza-6,9-diazonia(5,2,5,2)dispirohexadecane dichloride (dispirotripiperazine) were synthesized. To reveal relationships between chemical structure and antiviral activity, the compounds were characterized by fast atom bombardment mass, nuclear magnetic resonance, infra red spectroscopy, and elemental analysis and examined for cytotoxicity, inhibition of cell growth and antiviral activity under in vitro conditions. The results of this study demonstrate an excellent compatibility of the test compounds for confluent as well as proliferating cells and a potent structure-dependent inhibition of herpes simplex virus type 1 replication when added during viral adsorption. Functional group analysis revealed that both the dispirotripiperazine as well as the pyrimidine ring with a nitro group in the 5 position are necessary for activity. A reduction of electron density in the terminal pyrimidine rings enhanced the antiviral activity whereas electron donor substitutions reduced it. Introduction of a methyl group in position 2 of the pyrimidine had no influence on cytotoxicity or antiviral activity.


Subject(s)
Antiviral Agents/pharmacology , Herpesvirus 1, Human/drug effects , Piperazines/pharmacology , Spiro Compounds/pharmacology , Animals , Antiviral Agents/chemical synthesis , Antiviral Agents/toxicity , Cell Line , Chlorocebus aethiops , Cytopathogenic Effect, Viral/drug effects , Dogs , Dose-Response Relationship, Drug , HeLa Cells , Herpesvirus 1, Human/physiology , Humans , Piperazines/chemical synthesis , Piperazines/toxicity , Spiro Compounds/chemical synthesis , Spiro Compounds/toxicity , Structure-Activity Relationship , Virus Replication/drug effects
12.
J Mol Cell Cardiol ; 33(9): 1615-26, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11549341

ABSTRACT

Coxsackievirus B3 (CVB3)-induced myocarditis in NMRI mice represents a model for studying the pathogenesis of this chronic heart disease. Previously, we reported on specific cytokine patterns during the acute stage of myocarditis since cytokines are thought to play the important role in this cardiomyopathy. In this study, the expression of various cytokine mRNAs and CVB3-RNA kinetics was examined with particular emphasis on the late phase of myocarditis, by using reverse transcriptase-polymerase chain reaction (RT-PCR), in situ hybridization (ISH) and immunohistochemistry (IHC). In addition, replicating and persisting CVB3-RNAs were semiquantified by PCR-ELISA. Distinct histopathological changes responsible for ongoing heart disease were found and characterized by increased fibrosis, persistent cellular infiltration and degenerated necrotic myocytes. One of the most important findings of this study was that the mRNA-expression of TNF- alpha, IL-1 alpha, interferon- gamma, IL-10, IL-18, macrophage inflammatory protein-1 alpha (MIP-1 alpha), transforming growth factor- beta (TGF- beta) and inducible nitric oxide synthase (iNOS) persisted as long as 98 days after the virus infection. The induction of IL-10 as well as IFN- gamma mRNAs was also verified by ISH and IHC at days 28 and 98 p.i. The clearly apparent persistence of the viral genomes in the myocardium of infected mice was confirmed by seminested PCR, ISH, and PCR-enzyme linked immunoabsorbent assay (ELISA), showing the highest amount of viral RNA in myocardial cells at day 7 after infection. These data indicate that the persistence of viral RNA is associated with persistently high levels of cytokine mRNAs which, when translated, could severely contribute to pathological changes and injury of connective tissue in the chronic stage of myocarditis.


Subject(s)
Coxsackievirus Infections/immunology , Cytokines/biosynthesis , Enterovirus B, Human/physiology , Myocarditis/immunology , Myocardium/immunology , Animals , Chronic Disease , Coxsackievirus Infections/pathology , Coxsackievirus Infections/virology , Cytokines/genetics , Enterovirus B, Human/genetics , Enzyme-Linked Immunosorbent Assay , Heart/virology , Immunohistochemistry , In Situ Hybridization , Male , Mice , Mice, Inbred Strains , Myocarditis/chemically induced , Myocarditis/pathology , Myocarditis/virology , Myocardium/pathology , Polymerase Chain Reaction , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Viral/genetics , RNA, Viral/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
13.
J Basic Microbiol ; 41(3-4): 179-83, 2001.
Article in English | MEDLINE | ID: mdl-11512450

ABSTRACT

Surface cultures of Fusarium culmorum JP15 were found to respond to extracts of other fungi by enhanced production of orange-red fusarubin pigments and formation of aerial mycelium. Two inducers from strain Ulocladium sp. HKI 0226, the new (-)-terpestacin (1) and L-tenuazonic acid (2), were isolated. 1 inhibited syncytium formation by cells infected with respiratory syncytial virus (RSV).


Subject(s)
Bridged Bicyclo Compounds/metabolism , Fusarium/growth & development , Pigments, Biological/biosynthesis , Tenuazonic Acid/metabolism , Culture Media , Mass Spectrometry/methods , Morphogenesis , Respiratory Syncytial Viruses/physiology
14.
J Med Virol ; 64(4): 487-98, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11468734

ABSTRACT

Coxsackievirus B3 (CVB3) causes acute and chronic myocarditis, which is accompanied by an intense mononuclear leukocyte infiltration. Because myocardial tissue damage may either result from viral infections or from a dysregulated immune response, the susceptibility of human monocytes and macrophages to CVB3 was examined in this study with regard to virus replication, virus persistence, and release of cytokines. Monocytes were infected by CVB3 as shown by the intracellular appearance of plus- and minus-strand viral RNA, which was also capable of persisting for more than 10 days. Fresh monocytes were not permissive for full virus replication whereas monocyte-derived macrophages yielded a low amount of new viruses, which led to cell death. Although CVB3 infection induced the mRNA for the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1, and IL-6, only little cytokine production occurred. When infected monocytes were stimulated in addition by lipopolysaccharides (LPS), cytokine production was partially suppressed. In striking contrast, IL-10 expression was strongly and persistently induced by CVB3 on the mRNA and the protein level. These data show a dysregulated cytokine response in CVB3-exposed human monocytes and macrophages, which is characterized by a suppression of proinflammatory cytokines and a dominance of IL-10. This viral strategy may aid CVB3, causing chronic myocardiopathy.


Subject(s)
Coxsackievirus Infections/immunology , Enterovirus B, Human , Interleukin-10/biosynthesis , Monocytes/physiology , Cell Death , Cell Differentiation , Cells, Cultured , Coxsackievirus Infections/virology , Cytokines/biosynthesis , Cytokines/genetics , HeLa Cells , Humans , Interleukin-10/genetics , Lipopolysaccharides/pharmacology , Macrophages/immunology , Macrophages/virology , Monocytes/drug effects , Monocytes/virology , RNA, Messenger , RNA, Viral/analysis , Time Factors , Virus Replication
15.
J Virol Methods ; 95(1-2): 133-43, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11377720

ABSTRACT

In order to identify new potential antiviral drugs, small amounts of extracts or compounds have to be examined for cytotoxicity and antiviral activity in primary screening using a rapid, easy, inexpensive, and highly standardised test system. In this study, high-throughput cytopathic effect (CPE) inhibitory assays were established for coxsackie virus B3 on HeLa Ohio cells, influenza virus A on Madin-Darby canine kidney cells, and herpes simplex virus type 1 (HSV-1) on green monkey kidney cells that meet these requirements. The cytotoxic and the antiviral effects were quantified using a crystal violet uptake assay allowing automated handling of large numbers of candidate agents. To ensure comparable results with plaque reduction assays, the 50 and 90% plaque inhibitory concentrations of guanidine, amantadine, and phosphonoformic acid were used to standardise the anti-coxsackie virus B3, anti-influenza virus A, and anti-HSV-1 tests, respectively. The strong correlation between the antiviral activity determined by CPE-inhibitory assays and plaque reduction assay was further proved for other antivirals. In summary, low amounts of large numbers of compounds may be tested inexpensively and standardised within 24 h (coxsackie virus B3 and influenza virus A) or 48 h (herpes simplex virus type 1) post-infection using CPE inhibitory assays.


Subject(s)
Antiviral Agents/pharmacology , Enterovirus B, Human/drug effects , Herpesvirus 1, Human/drug effects , Influenza A virus/drug effects , Amantadine , Animals , Cell Division , Cell Line , Chlorocebus aethiops , Cytopathogenic Effect, Viral , Dogs , Enterovirus B, Human/pathogenicity , Foscarnet , Guanidine , HeLa Cells , Herpesvirus 1, Human/pathogenicity , Humans , Influenza A virus/pathogenicity , Reproducibility of Results , Staining and Labeling/methods , Time Factors , Viral Plaque Assay
16.
Virology ; 275(1): 77-88, 2000 Sep 15.
Article in English | MEDLINE | ID: mdl-11017789

ABSTRACT

The coxsackievirus B3 (CVB3) strain Nancy P establishes a persistent carrier-state infection without visible cytopathic effect in primary human fibroblasts (HuFi H), whereas the derivative variant PD induces a complete lysis of the cell monolayer. To define the molecular basis of this exceptional growth property, the complete genomes of both viruses were sequenced and compared to all published sequences of CVB3. As a result, six unique amino acid substitutions in the VP1 capsid protein were observed. Via hybrid virus construction, the lytic phenotype was transferred to a nonlytic cDNA-generated CVB3. Mapping experiments indicate that the presence of amino acid residues K78, A80, A91, and I92 in VP1 is sufficient to induce "lytic" infections in HuFi H cells. Binding assays demonstrate that CVB3 Nancy P preferentially binds to the human coxsackievirus-adenovirus receptor (CAR), while PD exhibits a very weak interaction with CAR but strong binding to the decay accelerating factor (DAF). These results suggest that the mutated amino acid residues in VP1 are involved in receptor recognition/binding. Moreover, the lytic replication of CVB3 PD and the hybrid virus in various nonpermissive rodent cell lines indicates that cell surface molecules other than CAR and DAF may be involved in attachment of this variant to cell surfaces.


Subject(s)
Capsid/metabolism , Enterovirus B, Human/metabolism , Enterovirus B, Human/pathogenicity , Genetic Variation/genetics , Amino Acid Substitution/genetics , Animals , Antibodies/pharmacology , Binding Sites , CD55 Antigens/metabolism , Capsid/chemistry , Capsid/genetics , Cell Line , Cells, Cultured , Coxsackie and Adenovirus Receptor-Like Membrane Protein , Cricetinae , Cytopathogenic Effect, Viral , DNA Mutational Analysis , DNA, Recombinant/genetics , Enterovirus B, Human/classification , Enterovirus B, Human/genetics , Fibroblasts/drug effects , Fibroblasts/pathology , Fibroblasts/virology , Humans , Mice , Models, Molecular , Mutation/genetics , Organ Specificity , Phenotype , Polymorphism, Genetic/genetics , Protein Binding/drug effects , Protein Conformation , Receptors, Virus/metabolism , Virus Replication/drug effects
17.
Radiologe ; 40(7): 632-9, 2000 Jul.
Article in German | MEDLINE | ID: mdl-10955294

ABSTRACT

PURPOSE: To compare spiral and conventional CT in the staging of carcinomas of the oral cavity, the oropharynx and hypopharynx. METHOD: Retrospectively 101 conventional CTs and prospectively 107 spiral CTs were analyzed regarding to correct T and N staging. CT results were compared with histological staging. RESULTS: In conventional CT, there were correctly staged 85% of T stages (T1 62%, T2 74%, T3 81%, T4 94%) and 85% of N stages (N0 79%, N1 71%, N2 89%, N3 94%). Spiral CT showed correct results in 84% of T stages (T1 67%, T2 74%, T3 88%, T4 95%) and in 86% of N stages (N0 82%, N1 78%, N2 90%, N3 93%). No statistically significant differences could be found between both CT methods. CONCLUSION: In spite of the tendency of improved diagnosis of T1, N0 and N1 stages no clear improvement in the staging of carcinomas of the oral cavity, the oropharynx and hypopharynx could be expected by spiral CT.


Subject(s)
Carcinoma/diagnostic imaging , Hypopharyngeal Neoplasms/diagnostic imaging , Mouth Neoplasms/diagnostic imaging , Oropharyngeal Neoplasms/diagnostic imaging , Tomography, X-Ray Computed/methods , Adult , Aged , Aged, 80 and over , Carcinoma/pathology , Female , Humans , Hypopharyngeal Neoplasms/pathology , Male , Middle Aged , Mouth Neoplasms/pathology , Neoplasm Staging , Oropharyngeal Neoplasms/pathology , Preoperative Care , Prospective Studies , Reproducibility of Results , Retrospective Studies , Tomography, X-Ray Computed/statistics & numerical data
18.
J Med Virol ; 61(4): 518-26, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10897072

ABSTRACT

Since cytokines play an important role in the pathogenesis of virus-induced chronic heart diseases, cytokine mRNA expression was studied in coxsackievirus B3-infected NMRI mice during the acute phase of myocarditis until the onset of chronic cardiac disease. Virus replication, cytokine induction, inflammatory cell infiltration and myocardial damage were studied by titer determination, reverse transcription-polymerase chain reaction (RT-PCR), and histopathology. To investigate whether the coxsackievirus B3-induced cytokine mRNA accumulation was only limited to the heart or generalized, spleen and thymus specimens were also included. Surprisingly, interleukin (IL)-10 as a deactivator of T cell and macrophage functions was transcribed in the myocardium nearly in parallel with virus replication from Day 1 through Day 14. At Day 3 p.i., the mRNA of IL-1alpha, tumor necrosis factor (TNF)-alpha, IL-6, and interferon (IFN)-beta accumulated. At Days 4, 7, and 14, IL-12-specific mRNA was produced. Furthermore, increasing amounts of IFN-gamma mRNA were found, whereas IL-2 and IL-4 mRNA remained undetectable. TNF-alpha, IL-1alpha, IL-10, IL-12, and IFN-gamma mRNA persisted into the late stage of myocarditis. In the spleen a closely correlated expression of virus and IL-10-specific mRNAs was also found, and in addition, IFN-beta, TNF-alpha, and IL-6 were detected. In striking contrast to heart and spleen tissue, the distinct expression of viral RNA in the thymus was not accompanied by an increased cytokine mRNA production. These data provide evidence for a unique coxsackievirus B3-induced cytokine pattern in the myocardium and spleen and suggest that persistently expressed IL-10 may play a leading role in acute and chronic myocarditis by subverting the immune response.


Subject(s)
Coxsackievirus Infections/immunology , Cytokines/analysis , Enterovirus , Myocarditis/immunology , Myocardium/immunology , Spleen/immunology , Thymus Gland/immunology , Acute Disease , Animals , Coxsackievirus Infections/pathology , Disease Models, Animal , Enterovirus/isolation & purification , Heart/virology , Interferon-beta/analysis , Interferon-gamma/analysis , Interleukins/analysis , Male , Mice , Myocarditis/pathology , Myocarditis/virology , Myocardium/pathology , Polymerase Chain Reaction , RNA, Messenger/analysis , Tumor Necrosis Factor-alpha/analysis
19.
J Hum Virol ; 2(6): 369-79, 1999.
Article in English | MEDLINE | ID: mdl-10774554

ABSTRACT

OBJECTIVES: The pathogenesis of coxsackievirus B3 (CVB3)-induced myocarditis was investigated in adult Han:NMRI mice. The outbred model, in comparison with inbred models, represents better the natural variable susceptibility of the human population. STUDY DESIGN/METHODS: We analyzed the replicating virus titer, the antibody response in the acute and chronic phase of disease, the histology of myocardial injury, and the persistence of viral RNA. RESULTS: NMRI mice infected with 5000 plaque-forming units (PFU) of the CVB3 variant "P"D, a lytic variant to human fibroblast lines, showed a peak of virus replication at day 14 and developed a severe acute myocarditis. The chronic myocarditis was characterized by progressive fibrosis, small foci of infiltrates, persistent viral RNA in the heart, and detectable anti-CVB3 IgG production and neutralizing antibody response up to day 98 postinfection. CONCLUSIONS: CVB3"P"D is able to induce chronic myocarditis in NMRI mice. This model provides a method for examining and proving the mechanisms of myocardial pathogenesis and of developing therapeutic strategies.


Subject(s)
Coxsackievirus Infections , Disease Models, Animal , Enterovirus B, Human , Myocarditis/virology , Animals , Antibodies, Viral/blood , Chronic Disease , Coxsackievirus Infections/pathology , Coxsackievirus Infections/virology , Enterovirus B, Human/immunology , Enterovirus B, Human/isolation & purification , Enterovirus B, Human/physiology , Heart/virology , Humans , Male , Mice , Myocarditis/pathology , Myocardium/pathology , Pancreas/virology , RNA, Viral/analysis , Virus Replication
20.
Zentralbl Bakteriol ; 284(2-3): 443-56, 1996 Jul.
Article in English | MEDLINE | ID: mdl-8837403

ABSTRACT

In our previous paper (29) we could show, that the replication of four Coxsackie B3 virus strains in human fibroblast lines from different origin was dependent on both the virus strain and the cell line used. Although generally no cytopathic effect could be observed out of one virus strain more pathogenic virus variants could be selected able to destroy virus-sensitive as well as insensitive fibroblasts. The present study was designed to characterize these virus strains by using several in vitro genetic markers. Differences were found concerning plaque size and the temperature marker, whereas the other markers (d, DD, DEAE-D) remained constant. Furthermore, these models of persistent as well as lytic CVB3 infection were analysed by immunoelectron microscopy to study the interaction of viral ligands with cellular receptors. The qualitative and quantitative differences in adsorption of the CVB3 strains to two human fibroblast lines as well as to HeLa cells corresponded well with the virological results. They underline that even in vitro in human cell lines of different origin changes in distribution, quantity and quality of receptors were demonstrable forming the base for the various virus sensitivity.


Subject(s)
Enterovirus B, Human/isolation & purification , Fibroblasts/virology , Receptors, Virus/metabolism , Animals , Cell Line , Enterovirus B, Human/genetics , Fetus/cytology , Fibroblasts/cytology , Genetic Markers , HeLa Cells , Humans , Immunohistochemistry , Microscopy, Immunoelectron , Rabbits
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