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  • 2008  (9)
  • Cinatl, J.  (9)
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  • 2008  (9)
  • 1
    In: Journal of Clinical Microbiology, 2008, Vol. 46(6), p.2122
    Description: Here we describe for the first time the productive in vitro infection of human retinal pigment epithelial cells by varicella-zoster virus (VZV), resulting in a typical cytopathic effect (CPE) that is characterized by enlarged cells with increased granularity. Depending on the CPE dissemination, high titers of up to 1.6 x 10 super(6) PFU of cell-free and cryostable VZV/ml can be recovered.
    Keywords: Retinal Pigment Epithelium ; Retina ; Replication ; Pigments ; Infection ; Varicella-Zoster Virus ; Replication;
    ISSN: 0095-1137
    ISSN: 00951137
    E-ISSN: 1098660X
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  • 2
    Language: English
    In: PLoS ONE, 2008, Vol.3(12), p.e3942
    Description: The hemagglutinin (HA) glycoprotein is the principal target of protective humoral immune responses to influenza virus infections but such antibody responses only provide efficient protection against a narrow spectrum of HA antigenic variants within a given virus subtype. Avian influenza viruses such as H5N1 are currently panzootic and pose a pandemic threat. These viruses are antigenically diverse and protective strategies need to cross protect against diverse viral clades. Furthermore, there are 16 different HA subtypes and no certainty the next pandemic will be caused by an H5 subtype, thus it is important to develop prophylactic and therapeutic interventions that provide heterosubtypic protection. ; Here we describe a panel of 13 monoclonal antibodies (mAbs) recovered from combinatorial display libraries that were constructed from human IgM memory B cells of recent (seasonal) influenza vaccinees. The mAbs have broad heterosubtypic neutralizing activity against antigenically diverse H1, H2, H5, H6, H8 and H9 influenza subtypes. Restriction to variable heavy chain gene IGHV1-69 in the high affinity mAb panel was associated with binding to a conserved hydrophobic pocket in the stem domain of HA. The most potent antibody (CR6261) was protective in mice when given before and after lethal H5N1 or H1N1 challenge. ; The human monoclonal CR6261 described in this study could be developed for use as a broad spectrum agent for prophylaxis or treatment of human or avian influenza infections without prior strain characterization. Moreover, the CR6261 epitope could be applied in targeted vaccine strategies or in the design of novel antivirals. Finally our approach of screening the IgM memory repertoire could be applied to identify conserved and functionally relevant targets on other rapidly evolving pathogens.
    Keywords: Research Article ; Biotechnology ; Immunology -- Immunity To Infections ; Immunology -- Innate Immunity ; Microbiology -- Innate Immunity ; Virology -- Antivirals, Including Modes Of Action And Resistance ; Virology -- New Therapies, Including Antivirals And Immunotherapy ; Infectious Diseases -- Respiratory Infections ; Infectious Diseases -- Viral Infections ; Respiratory Medicine -- Respiratory Infections
    E-ISSN: 1932-6203
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  • 3
    Language: English
    In: Clinical cancer research : an official journal of the American Association for Cancer Research, 15 October 2008, Vol.14(20), pp.6531-7
    Description: Neuroblastomas frequently show expression of the epidermal growth factor receptor (EGFR) and may therefore be susceptible to EGFR-targeted therapies. Here, EGFR expression and functionality was investigated in parental chemosensitive neuroblastoma cell lines (UKF-NB-3, IMR-32, NLF, SH-SY5Y) and their cisplatin-resistant sublines (UKF-NB-3(r)CDDP(1000), IMR-32(r)CDDP(1000), NLF(r)CDDP(1000), and SH-SY5Y(r)CDDP(500)). Moreover, the EGFR antibody cetuximab, the EGFR tyrosine kinase inhibitor Tyrphostin B46, and recombinant EGFR-targeted toxins were investigated for their influence on the viability and growth of neuroblastoma cells. EGFR expression and function was measured by flow cytometry or Western blot. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was examined by immunostaining for active caspase-3 or cleaved poly(ADP-ribose) polymerase. Cellular binding of FITC-labeled immunotoxins was studied by flow cytometry, and cellular uptake was studied by confocal laser scanning microscopy. The EGFR-targeted antibody and growth factor toxins scFv(14E1)- Pseudomonas exotoxin A (ETA) and TGF-alpha-ETA exerted anti-cancer effects in neuroblastoma cell lines that were insensitive to cetuximab or EGFR tyrosine kinase inhibitors. Furthermore, adaptation of chemosensitive neuroblastoma cells to cisplatin increased EGFR expression and sensitivity to both recombinant toxins. Treatment of chemosensitive neuroblastoma cells with cisplatin reversibly increased EGFR expression, whereas cisplatin-resistant cells showed enhanced EGFR expression independent of the presence of cisplatin. Combination treatment with scFv(14E1)-ETA or TGF-alpha-ETA and cisplatin exerted significantly improved anticancer effects compared with either single treatment in parental neuroblastoma cells, cisplatin-resistant sublines, and primary cultures. EGFR-targeted cytotoxic reagents such as scFv(14E1)-ETA and TGF-alpha-ETA represent promising candidates for further development as antineuroblastoma agents, especially in combination with cisplatin.
    Keywords: Drug Resistance, Neoplasm ; Adp Ribose Transferases -- Therapeutic Use ; Antineoplastic Agents -- Pharmacology ; Bacterial Toxins -- Therapeutic Use ; Cell Proliferation -- Drug Effects ; Cisplatin -- Pharmacology ; Erbb Receptors -- Antagonists & Inhibitors ; Exotoxins -- Therapeutic Use ; Neuroblastoma -- Drug Therapy ; Virulence Factors -- Therapeutic Use
    ISSN: 1078-0432
    E-ISSN: 15573265
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  • 4
    In: Cardiovascular Research, 2008, Vol. 77(3), pp.544-550
    Description: AIMS: The endothelium represents a natural site of human cytomegalovirus (HCMV) infection involved in viral spreading and persistence. Moreover, HCMV infection of endothelial cells has been associated with different pathological conditions of the cardiovascular system. Here, the influence of the antiepileptic drug valproic acid (VPA) was investigated on HCMV replication in human umbilical vein endothelial cells alone or in combination with the antiviral drugs ganciclovir, cidofovir or foscarnet.METHODS AND RESULTS: HCMV replication was observed by immunostaining for viral antigens and by virus yield assay. Protein expression and phosphorylation were examined by western blot. Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye reduction assay. Therapeutic VPA concentrations (〈 or =1 mM) increased HCMV immediate early antigen, late antigen, and viral titres of different endotheliotropic and non-endotheliotropic HCMV strains in a concentration- and time-dependent manner up to 30-fold. Moreover, VPA impaired the antiviral activity of the anti-HCMV drugs ganciclovir, cidofovir, and foscarnet. VPA inhibits histone deacetylases (HDAC) and induces HDAC-independently extracellular signal-regulated kinases 1/2 (ERK 1/2) phosphorylation in endothelial cells. Both effects observed, HCMV stimulation and interference with antiviral drugs, depend on HDAC inhibition but not on ERK 1/2 phosphorylation.CONCLUSION: These findings suggest to carefully monitor the frequency of HCMV reactivation in cardiovascular patients treated with VPA (or other HDAC inhibitors) in comparison to control individuals.
    Keywords: Human Cytomegalovirus ; Antiviral Therapy ; Endothelium
    ISSN: 0008-6363
    E-ISSN: 1755-3245
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  • 5
    Language: English
    In: Neoplasia, December 2008, Vol.10(12), pp.1402-1410
    Description: Prolonged treatment of leukemic cells with chemotherapeutic agents frequently results in development of drug resistance. Moreover, selection of drug-resistant cell populations may be associated with changes in malignant properties such as proliferation rate, invasiveness, and immunogenicity. In the present study, the sensitivity of cytarabine (1-β- -arabinofuranosylcytosine, araC)-resistant and parental human leukemic cell lines (T-lymphoid H9 and acute T-lymphoblastic leukemia Molt-4) to natural killer (NK) cell-mediated killing was investigated. The results obtained demonstrate that araC-resistant H9 and Molt-4 (H9 ARAC and Molt-4 ARAC ) cell lines are more sensitive to NK cell-mediated lysis than their respective parental cell lines. This increased sensitivity was associated with a higher surface expression of ligands for the NK cell-activating receptor NKG2D, notably UL16 binding protein-2 (ULBP-2) and ULBP-3 in H9 ARAC and Molt-4 ARAC cell lines. Blocking ULBP-2 and ULBP-3 or NKG2D with monoclonal antibody completely abrogated NK cell lysis. Constitutive phosphorylated extracellular signal-regulated kinase (ERK) but not pAKT was higher in araC-resistant cells than in parental cell lines. Inhibition of ERK using ERK inhibitor PD98059 decreased both ULBP-2/ULBP-3 expression and NK cell cytotoxicity. Furthermore, overexpression of constitutively active ERK in H9 parental cells resulted in increased ULBP-2/ULBP-3 expression and enhanced NK cell lysis. These results demonstrate that increased sensitivity of araC-resistant leukemic cells to NK cell lysis is caused by higher NKG2D ligand expression, resulting from more active ERK signaling pathway.
    Keywords: Medicine
    ISSN: 1476-5586
    E-ISSN: 1476-5586
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  • 6
    Language: English
    In: Phytomedicine, 2008, Vol.15(1), pp.79-83
    Description: are widely used as traditional medicine in Lebanon for treatment of different infection diseases. In the present study we reported the phytochemical composition analyzed by GC–MS of wood essential oil and cones and leaves ethanol extracts. The main components of wood essential oil were himachalol (22.50%), -himachalene (21.90%), and -himachalene (10.50%). Leaves ethanol extract was characterized by a high content of germacrene (29.40%). The same extract obtained from cones essentially contained -pinene (51.0%) and -myrcene (13.0%). Moreover, we investigated extracts, essential oil, and identified compounds for their antiviral activities against herpes simplex virus type 1 (HSV-1). Cytotoxicity was evaluated by MTT assay in Vero cells. Cones and leaves ethanol extracts exhibited an interesting activity with IC of 0.50 and 0.66 mg/ml, respectively, at non-cytotoxic concentration. A comparable activity was found when essential oil was tested (IC of 0.44 mg/ml).
    Keywords: Cedrus Libani A. Rich ; Antiviral Activity ; Herpes Simplex Virus (Hsv-1) ; Gc–MS Analysis ; Medicine ; Pharmacy, Therapeutics, & Pharmacology
    ISSN: 0944-7113
    E-ISSN: 1618-095X
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  • 7
    In: Annals of Occupational Hygiene, 2008, 2008, Vol. 52(7), pp.615-622
    Description: Objectives: Our paper measures the prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV) in patients at the University Hospital of Frankfurt/Main, and correlates the prevalence with risk factors for exposure to and infection of healthcare workers (HCWs). Individual risk assessments were calculated for exposed HCWs. Methods: Survey of patients admitted to a German University Hospital. Markers for HBV, HCV and HIV were studied and evaluated statistically. Data on needlestick injuries (NSIs) among HCWs were correlated with the prevalence of infectious patients. Results: The HBV, HCV and HIV prevalence among patients at the University Hospital were 5.3% ( n  = 709/13 358), 5.8% ( n  = 1167/20 163) and 4.1% ( n  = 552/13 381), respectively. Our results indicate that the prevalence of blood-borne infections in patients was about nine times higher for HBV, ∼15 times higher for HCV and ∼82 times higher for HIV than in the overall German population. The highest risk of acquiring a blood-borne infection via NSI was found in the department of internal medicine due to increased prevalence of blood-borne pathogens in patients under treatment. Conclusions: While accidental NSIs were most frequent in surgery, the nominal risk of blood-borne virus infection was greatest in the field of internal medicine. The study underlines the importance of HBV vaccinations and access to HIV-post-exposure prophylaxis for HCWs as well as the use of anti-needlestick devices.
    Keywords: Blood - Borne Viruses ; Healthcare Workers ; Occupational Infections
    ISSN: 0003-4878
    E-ISSN: 1475-3162
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  • 8
    Language: English
    In: Journal of Ethnopharmacology, 2008, Vol.120(1), pp.108-111
    Description: Severe acute respiratory syndrome (SARS) is a life-threatening disease caused by the SARS coronavirus (SARS-CoV). The development of new antiviral agents for SARS-CoV is an important issue. We tried to find potential resource from Traditional Chinese medicine (TCM) for development of new drugs against SARS-CoV. Our team recruited the potential TCM formulae (also known as Kampo) from two TCM books, Shang-Han Lun (Discussion of Cold-Induced Disorders) and Wen-Bing Tiau-Bein (Differential Management of Febrile Diseases). Several herbs, which were believed to be beneficial for SARS by experienced TCM doctors were also recruited. In addition, a vegetable polular in Taiwan, China and Malaysia, the tender leaf of Roem (also known as Cedrela sinensis, belongs to the family Meliacceae) was also recruited under the suggestion of botanic experts. These TCM products and plant extrats were then tested for the effectiveness against SARS-CoV in vitro. Only TSL-1, the extract from tender leaf of Roem was found to have an evident effect against SARS-CoV with selectivity index 12∼17. This paper reports for the first time that extract from a vegetable, the tender leaf of Roem, can inhibit SARS-CoV in vitro. Thererfore, the tender leaf of Roem may be an important resource agninst SARS-CoV.
    Keywords: Sars ; Herb ; Traditional Chinese Medicine (Tcm) ; Toona Sinensis Roem ; Pharmacy, Therapeutics, & Pharmacology
    ISSN: 0378-8741
    E-ISSN: 1872-7573
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  • 9
    Language: English
    In: Chemistry & Biodiversity, March 2008, Vol.5(3), pp.461-470
    Description: The chemical composition of the essential oils of ssp. ssp. , and was determined by GC/MS analysis. Essential oils have been evaluated for their inhibitory activity against SARS‐CoV and HSV‐1 replication by visually scoring of the virus‐induced cytopathogenic effect post‐infection. oil exerted an interesting activity against SARS‐CoV with an value of 120 μg/ml and a selectivity index (SI) of 4.16. This oil was characterized by the presence of ‐ocimene, 1,8‐cineole, ‐pinene, and ‐pinene as the main constituents. ssp. oil, in which ‐pinene and ‐myrcene were the major constituents, revealed antiviral activity against HSV‐1 with an value of 200 μg/ml and a SI of 5.
    Keywords: Severe Acute Respiratory Syndrome Sars ; Antiviral Activity ; Cytotoxic Activity ; Essential Oils ; Laurus Nobilis ; Ssp. ; Thuja Orientalis ; Ssp. ; Pistacia Palaestina ; Salvia Officinalis ; Satureja Thymbra
    ISSN: 1612-1872
    E-ISSN: 1612-1880
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