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1.
Ann Fr Anesth Reanim ; 32(12): e199-203, 2013 Dec.
Article in French | MEDLINE | ID: mdl-24209991

ABSTRACT

The technology of anesthesia ventilators has substantially progressed during last years. The choice of a pediatric anesthesia ventilator needs to be led by multiple parameters: requirement, technical (pneumatic performance, velocity of halogenated or oxygen delivery), cost (purchase, in operation, preventive and curative maintenance), reliability, ergonomy, upgradability, and compatibility. The demonstration of the interest of pressure support mode during maintenance of spontaneous ventilation anesthesia makes this mode essential in pediatrics. In contrast, the financial impact of target controlled inhalation of halogenated has not be studied in pediatrics. Paradoxically, complex and various available technologies had not been much prospectively studied. Anesthesia ventilators performances in pediatrics need to be clarified in further clinical and bench test studies.


Subject(s)
Anesthesiology/instrumentation , Pediatrics/instrumentation , Ventilators, Mechanical , Anesthesia/methods , Anesthetics, Inhalation/administration & dosage , Child , Equipment Design , Humans , Intermittent Positive-Pressure Ventilation , Ventilators, Mechanical/economics
2.
Minerva Med ; 103(4): 293-7, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22805621

ABSTRACT

AIM: We compared Hepatitis C virus (HCV) genotyping by direct sequencing of the non-structural 5b region (NS5b) and a commercial PCR/hybridization method based on the conserved 5´-untranslated region (5'UTR). METHODS: One hundred twenty HCV containing plasma samples were analyzed by NS5b sequencing with focus on samples with undetermined results or 1b subtype identification in the used combination of Cobas® AmpliPrep/Cobas® TaqMan96® PCR and subsequent Versant® HCV Genotype 2.0 Assay (LiPA). RESULTS: There was 100% concordance between the two methods for genotyping but only 83% for subtyping. Seventeen samples were designated 1b by hybridization but subtype 1a by NS5b sequencing. This is a general 5'UTR problem as the discordant results were additionally confirmed by 5'UTR sequencing. Thus our routine combination not only misclassified 38.6% of subtype 1a isolates as 1b but in contrast to NS5b sequencing was unable to discriminate between subtypes 2a/c, or 4a/c/d and also failed on a newly described subtype (10a/3k). [corrected]. CONCLUSIONS: [corrected] The applied 5'UTR methods allow the rapid determination of HCV genotypes but failed to correctly identify the subtype in many samples. This has implications for epidemiological studies or forensic evaluation of chains of infection and NS5b sequencing therefore is our method of choice under those circumstances.


Subject(s)
5' Untranslated Regions/genetics , Genotyping Techniques/methods , Hepacivirus/genetics , Comparative Genomic Hybridization/methods , Genotype , Humans , Polymerase Chain Reaction/methods , Sequence Analysis, DNA/methods
3.
Clin Microbiol Infect ; 17(2): 181-9, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20345467

ABSTRACT

The emergence of metallo-ß-lactamase (MBL)-producing Enterobacteriaceae is a serious public health concern. Producers have been repeatedly isolated from patients and long-term care facility (LTCF) residents around Bolzano, and we sought to assess their prevalence and clinical impact. All routine Enterobacteriaceae isolates from a Bolzano tertiary-care hospital and associated long-term care facilities in 2008 (n = 5500) were screened for MBLs, with case details reviewed for the source patients. In total, 36 producers were obtained from 29 patients, comprising 14 Escherichia coli, six Klebsiella pneumoniae, four Klebsiella oxytoca, four Citrobacter freundii, two Enterobacter cloacae and two Morganella morganii, as well as single Citrobacter amalonaticus, Enterobacter aerogenes, Providencia stuartii and Proteus mirabilis isolates. All were PCR-positive for bla(VIM) and 25 were PCR-positive for qnrS; 19 non-K. pneumoniae had bla(SHV) and one had bla(CTX-M-group1); 13 were from 12 LTCF residents and 23 were from 17 acute-care patients. All these patients had serious underlying diseases with prolonged hospitalization or LTCF stay; only seven had infections due to the MBL producers, comprising four urinary tract infections, two catheter-related bloodstream infections and one patient with both a surgical site infection and pneumonia. Five patients had more than one MBL-producing organism. Pulsed-field gel electrophoresis identified a cluster of six related E. coli, whereas pairs of K. pneumoniae and C. freundii isolates had >85% profile similarity. Transformants prepared from two isolates were shown to be PCR-positive for bla(VIM), qnrS and bla(SHV); their plasmids gave similar restriction fragment length polymorphism patterns, and bla(VIM-1), qnrS1 and bla(SHV-12) were detected by sequencing.


Subject(s)
Cross Infection/microbiology , Enterobacteriaceae Infections/microbiology , Enterobacteriaceae/enzymology , Enterobacteriaceae/isolation & purification , beta-Lactamases/genetics , Aged , Aged, 80 and over , Bacterial Typing Techniques , Child, Preschool , Cluster Analysis , Electrophoresis, Gel, Pulsed-Field , Enterobacteriaceae/classification , Enterobacteriaceae/genetics , Female , Hospitals , Humans , Infant , Infant, Newborn , Italy , Long-Term Care , Male , Middle Aged , Molecular Typing , Polymerase Chain Reaction
4.
Clin Microbiol Infect ; 16(7): 934-44, 2010 Jul.
Article in English | MEDLINE | ID: mdl-19686277

ABSTRACT

Long-term-care facilities (LTCFs) are reservoirs of resistant bacteria. We undertook a point-prevalence survey and risk factor analysis for specific resistance types among residents and staff of a Bolzano LTCF and among geriatric unit patients in the associated acute-care hospital. Urine samples and rectal, inguinal, oropharyngeal and nasal swabs were plated on chromogenic agar; isolates were typed by pulsed-field gel electrophoresis; resistance genes and links to insertion sequences were sought by PCR; plasmids were analysed by PCR, restriction fragment length polymorphism and incompatibility grouping. Demographic data were collected. Of the LTCF residents, 74.8% were colonized with ≥1 resistant organism, 64% with extended-spectrum ß-lactamase (ESBL) producers, 38.7% with methicillin-resistant Staphylococcus aureus (MRSA), 6.3% with metallo-ß-lactamase (MBL) producers, and 2.7% with vancomycin-resistant enterococci. Corresponding rates for LTCF staff were 27.5%, 14.5%, 14.5%, 1.5% and 0%, respectively. Colonization frequencies for geriatric unit patients were lower than for those in the LTCF. Both clonal spread and plasmid transfer were implicated in the dissemination of MBL producers that harboured IncN plasmids bearing bla(VIM-1), qnrS, and bla(SHV-12). Most (44/45) ESBL-producing Escherichia coli isolates had bla(CTX-M) genes of group 1; a few had bla(CTX-M) genes of group 9 or bla(SHV-5); those with bla(CTX-M-15) or bla(SHV-5) were clonal. Risk factors for colonization of LTCF residents with resistant bacteria included age ≥86 years, antibiotic treatment in the previous 3 months, indwelling devices, chronic obstructive pulmonary disease, physical disability, and the particular LTCF unit; those for geriatric unit patients were age and dementia. In conclusion, ESBL-producing and MBL-producing Enterobacteriaceae and MRSA were prevalent among the LTCF residents and staff, but less so in the hospital geriatric unit. Education of LTCF employees and better infection control are proposed to minimize the spread of resistant bacteria in the facility.


Subject(s)
Bacteria/isolation & purification , Drug Resistance, Multiple, Bacterial , Hospital Units , Long-Term Care , Patients , Personnel, Hospital , Bacteria/drug effects , Cross Infection/diagnosis , Cross Infection/epidemiology , Cross Infection/microbiology , Electrophoresis, Gel, Pulsed-Field , Enterobacteriaceae/drug effects , Enterobacteriaceae/isolation & purification , Enterococcus/drug effects , Enterococcus/isolation & purification , Health Services for the Aged , Hospitals , Humans , Inguinal Canal/microbiology , Italy , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Nose/microbiology , Oropharynx/microbiology , Plasmids , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Rectum/microbiology , Risk Factors , Urine/microbiology , Vancomycin Resistance
5.
J Prev Med Hyg ; 50(2): 117-20, 2009 Jun.
Article in English | MEDLINE | ID: mdl-20099443

ABSTRACT

BACKGROUND: The number of patients with severe Clostridium difficile-associated diarrhoea (CDAD) increases. Health care facilities are requested to establish rates of nosocomially acquired CDAD (N-CDAD) to understand the impact of control or prevention measures, and the burden of N-CDAD on health care resources. OBJECTIVE: Aim of the single-center surveillance project was to establish local prevalence rates of N-CDAD in adult acute care medical patients. METHODS: For a period of at least one year, all diarrhoeal stools from inpatients of a general internal medicine ward were tested for Clostridium difficile toxin A. Case record files were retrospectively analysed and questionnaires were completed for patients with positive stool assays who met the case definitions. RESULTS AND DISCUSSION: During the surveillance period, 2,610 medical patients had been acutely hospitalized. Stools had been submitted to the hospital laboratory from 163 patients (6.2%) because of diarrhoea and were screened for Clostridium difficile cytotoxin. Complete data sets were available for analysis from 150 patients. Of 137 identified potential cases, 77 (56.2%) met the case definitions for nosocomial diarrhoea. Thirteen of the patients with nosocomial diarrhoea (16.9%) were detected positive by the Clostridium difficile toxin A assay. The overall prevalence of N-CDAD among inpatients was 8.7 cases/100 diarrhoeal stools. The mean number ofN-CDAD cases was 62.3 cases/100,000 patient days and 5 cases/1,000 patient admissions. The mean age of N-CDAD patients was 79.4 years (range 71 to 92). All patients were given broad-spectrum antibiotics before acute diarrhoea developed. Four patients died for reasons not directly related to N-CDAD which confirms increased disease severity as an important risk factor. CONCLUSIONS: This single-center surveillance project, which established N-CDAD rates at frequencies currently reported from international surveys, is useful as benchmark and will help in understanding patterns and impact of N-CDAD at the regional level.


Subject(s)
Clostridioides difficile/isolation & purification , Cross Infection/epidemiology , Cross Infection/microbiology , Diarrhea/epidemiology , Diarrhea/microbiology , Enterocolitis, Pseudomembranous/epidemiology , Aged , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , Cross Infection/drug therapy , Diarrhea/drug therapy , Enterocolitis, Pseudomembranous/drug therapy , Enterocolitis, Pseudomembranous/microbiology , Feces/microbiology , Female , Hospitals, Teaching , Humans , Italy/epidemiology , Male , Prevalence , Retrospective Studies
6.
Eur J Clin Microbiol Infect Dis ; 25(11): 681-6, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17036151

ABSTRACT

The aim of this epidemiological study was to determine the prevalence of respiratory viruses, including new viruses, in hospitalised children in Austria. Two hundred fourteen nasopharyngeal samples from hospitalised children were tested for the presence of viruses using cell culture and PCR and/or viral antigen assays. The results revealed a parainfluenza virus 1 (PIV1) outbreak that ended right before the onset of the influenza season, with nearly no overlapping, moderate respiratory syncytial virus (RSV) activity, and only a few adenoviruses. Human metapneumovirus (hMPV) was present in 14.5% of the total samples but was detected in combination with other viruses in only five cases: with PIV1 in three cases and with RSV in two cases. There were no cases of dual infection with hMPV and flu or adenovirus. This suggests that hMPV alone is a leading cause of hospitalisation in children under 1 year of age. Interestingly, hMPV, in contrast to RSV, coincided with PIV1 but was absent during the community outbreak of the flu. Samples were also tested for Mimiviridae, a group of newly described DNA viruses that are similar to Legionella spp., replicate in water amoebae, and also have been found in alveolar cells. However, mimivirus was detected neither in respiratory samples nor in amoebae-containing water samples, indicating that this particular type of virus is either not abundant or does not contribute to paediatric respiratory illnesses.


Subject(s)
Hospitalization , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/virology , Virus Diseases/epidemiology , Virus Diseases/virology , Animals , Austria/epidemiology , Cell Line , Child, Preschool , Humans , Infant , Influenza A virus , Influenza, Human/epidemiology , Influenza, Human/virology , Metapneumovirus , Parainfluenza Virus 1, Human , Paramyxoviridae Infections/epidemiology , Paramyxoviridae Infections/virology , Polymerase Chain Reaction , Prevalence , Respirovirus Infections/epidemiology , Respirovirus Infections/virology , Virus Cultivation , Viruses/classification , Viruses/genetics , Viruses/isolation & purification
8.
Transplant Proc ; 37(4): 1839-44, 2005 May.
Article in English | MEDLINE | ID: mdl-15919482

ABSTRACT

BACKGROUND: In contrast to Epstein-Barr virus (EBV)-associated posttransplant lymphoproliferative disorders (PTLD), EBV-associated leiomyomatous tumors have thus far only rarely been described. CASE REPORT: Two years after heart transplantation with ATG induction, cyclosporine (CsA; trough levels of 250 ng/mL)-based triple drug immunosuppression), a 23-year-old patient developed a small round lesion within the left lateral liver segment. The patient underwent ultrasound-guided biopsy followed by liver resection. Histological and immunohistological examination showed a leiomyosarcoma. In situ hybridization using EBV-specific EB endoplasmic reticulum-RNA showed an intensive signal in almost all tumor cells. The tumor stained for EB nuclear antigen (EBNA)-2-protein. Immunosuppression was drastically reduced, namely, CsA levels <100 ng/dL, prednisolone 5 mg, azathioprine withdrawn, and antiviral chemotherapy initiated with 10 days of IV gancyclovir and acyclovir followed by oral famcyclovir. During the follow-up, anti-EBV-IgM, anti-early antigen antibodies, and anti-EBNA antibodies were continuously monitored excluding significant EBV replication. Eighteen months post-liver resection, and high-resolution computed tomography scan demonstrated two paravertebral tumors. These lesions and a small nodule at the left ankle were resected revealing identical leiomyosarcomata. Immunosuppression was further reduced (CsA levels 75 ng/dL) and famcyclovir maintenance therapy started. Nevertheless, 2 years later the patient again developed tumor recurrence (perirectal, liver, and right adrenal gland); the tumors were surgically removed. The therapy was switched to Rapamycin and famcyclovir was continued. Three years after the last surgical intervention, the patient is well and recurrence-free. CONCLUSION: Long-term survival in patients with posttransplant EBV-associated leiomyosarcoma can be achieved by combined surgical intervention, reduction of immunosuppression, switch to Sirolimus, and antiviral chemotherapy.


Subject(s)
Epstein-Barr Virus Infections/drug therapy , Heart Transplantation/methods , Leiomyosarcoma/surgery , Liver Neoplasms/surgery , Acyclovir/therapeutic use , Adult , Antiviral Agents/therapeutic use , Cardiomyopathy, Dilated/surgery , Ganciclovir/therapeutic use , Heart Transplantation/pathology , Humans , Leiomyosarcoma/diagnostic imaging , Liver Neoplasms/diagnostic imaging , Male , Microsurgery , Recurrence , Tomography, X-Ray Computed , Treatment Outcome , Ultrasonography
9.
Exp Dermatol ; 14(1): 41-9, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15660918

ABSTRACT

CD34(+) progenitor cells carrying human herpesvirus-8, Kaposi's sarcoma-associated herpesvirus (HHV-8/KSHV), have been described in the peripheral blood of AIDS patients suffering from Kaposi's sarcoma (KS). In this study, we investigated the influence of HHV-8 on the differentiation of CD34(+) progenitor cells. Native CD34(+) cells derived from cord blood could be infected by a laboratory strain of HHV-8, as shown by immunofluorescence staining and polymerase chain reaction, but no significant initial maturation/differentiation effects were observed. In addition, these infected cells were differentiated into immature and mature dendritic cells (DCs) using cytokine induction with recombinant human granulocyte-macrophage colony-stimulating factor (rhGm-CSF), recombinant human tumor necrosis factor (rhTNF-alpha) and recombinant human stem cell factor (rhSCF). Double immunofluorescence and flow cytometry studies demonstrated that virus infection did not impair the development of immature and mature DC populations. Subsequently, the immunostimulating capacity of DC populations was tested in a mixed lymphocyte reaction using allogeneic T-cells. The HHV-8-infected CD34(+) progenitor cell-derived mature DC population showed a significantly enhanced antigen-presenting capacity, compared to non-infected DCs, which was not observed with the immature DCs. This suggests stimulation of DC function by HHV-8 infection. Because there are only a small percentage of HHV-8-positive DCs in the preparations and because it is not clear whether infection is abortive or productive to some extent, this seems to be most likely due to an indirect viral effect.


Subject(s)
Antigens, CD34/immunology , Dendritic Cells/immunology , Herpesvirus 8, Human/immunology , Lymphocyte Activation/immunology , Stem Cells/immunology , Antibodies, Monoclonal/immunology , Antigens, CD/analysis , Cell Differentiation/drug effects , Cytokines/pharmacology , DNA, Viral/analysis , Dendritic Cells/ultrastructure , Dendritic Cells/virology , Fetal Blood/cytology , Herpesvirus 8, Human/genetics , Humans , Immunohistochemistry , Lymphocyte Culture Test, Mixed , Microscopy, Electron, Transmission , Stem Cells/chemistry , Stem Cells/virology , T-Lymphocytes/immunology
11.
Lancet ; 358(9288): 1157-9, 2001 Oct 06.
Article in English | MEDLINE | ID: mdl-11597674

ABSTRACT

HLA-A2-restricted T cells show peptide-specific activity against cytomegalovirus and leukaemia cells. We retrospectively analysed the influence of donor cytomegalovirus serostatus on the outcome of 103 consecutive patients who had leukaemia and who received bone-marrow transplants from HLA-identical sibling donors. We found that donor cytomegalovirus seropositivity significantly improved overall survival (p=0.02) as a result of lower relapse incidence (p=0.035) in HLA-A2-positive but not HLA-A2-negative recipients. In HLA-A2-positive recipients donor cytomegalovirus seropositivity was associated with chronic graft-versus-host disease (GVHD), but even in patients without chronic GVHD donor cytomegalovirus seropositivity significantly improved survival (p=0.0483). These preliminary data provide evidence that at least in HLA-A2-positive recipients, transplantation of bone marrow from cytomegalovirus positive, HLA-identical sibling donors seems to be associated with substantial graft-versus-leukaemia activity, and suggests a cross-reactivity of cytomegalovirus-specific donor-derived cytotoxic T cells with HLA-A2-restricted recipient minor histocompatibility antigens.


Subject(s)
Bone Marrow Transplantation , Cytomegalovirus/immunology , Leukemia/therapy , Nuclear Family , Adolescent , Adult , Child , Child, Preschool , Combined Modality Therapy , Female , HLA-A2 Antigen/isolation & purification , Humans , Infant , Leukemia/mortality , Male , Retrospective Studies , Survival Analysis
12.
J Infect Dis ; 183(9): 1388-94, 2001 May 01.
Article in English | MEDLINE | ID: mdl-11294671

ABSTRACT

Iron chelation therapy of Plasmodium falciparum infection alleviates the clinical course of cerebral malaria in children. This study assessed the underlying mechanisms of this therapy. Cytokine stimulation of human (intestinal cell line DLD-1) or murine cells (murine macrophage cell line RAW 264.7) resulted in increased nitric oxide (NO) formation and decreased survival of plasmodia within cocultured human erythrocytes. The addition of desferrioxamine (DFO) before cytokine treatment increased both NO formation and parasite killing but had no effect in the presence of the inhibitor of NO formation, L-N6-(1-iminoethyl)-lysine. Moreover, peroxynitrite, which is formed after chemical reaction of NO with superoxide, appears to be the principal effector molecule for macrophage-mediated cytotoxicity toward P. falciparum, and interferon-gamma is a major regulatory cytokine for this process. The effect of DFO on the clearance of plasmodia appears to be due to enhanced generation of NO, rather than to limitation of iron availability to the parasite.


Subject(s)
Deferoxamine/therapeutic use , Iron Chelating Agents/therapeutic use , Iron/metabolism , Malaria, Falciparum/immunology , Nitric Oxide/biosynthesis , Plasmodium falciparum/drug effects , Animals , Cells, Cultured , Coculture Techniques , Erythrocytes/immunology , Erythrocytes/parasitology , Humans , Interferon-gamma/immunology , Lipopolysaccharides/immunology , Macrophages/immunology , Macrophages/parasitology , Malaria, Falciparum/drug therapy , Mice , Nitric Oxide/toxicity , Plasmodium falciparum/immunology , Plasmodium falciparum/metabolism , RNA, Messenger/analysis , Superoxide Dismutase/metabolism , Tumor Cells, Cultured
13.
Scand J Infect Dis ; 33(3): 236-8, 2001.
Article in English | MEDLINE | ID: mdl-11303820

ABSTRACT

An unusual cutaneous relapse of visceral leishmaniasis (initially mistaken for eruptive histiocytomas) was seen in an AIDS patient despite good virological and CD4+ T-cell responses to highly active antiretroviral therapy. Splenectomy and the patient's low CD8+ T-cell count are discussed as possible causes of failed disease control.


Subject(s)
AIDS-Related Opportunistic Infections/parasitology , HIV Infections/drug therapy , Leishmaniasis, Visceral/etiology , AIDS-Related Opportunistic Infections/drug therapy , Adult , Antiretroviral Therapy, Highly Active , CD4 Lymphocyte Count , CD8-Positive T-Lymphocytes , Diagnosis, Differential , HIV Infections/complications , Hemophilia B/complications , Humans , Leishmaniasis, Visceral/drug therapy , Recurrence , Splenectomy , Treatment Failure
15.
J Invest Dermatol ; 116(1): 150-6, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11168811

ABSTRACT

The effect of herpes virus infection on human dermal microvascular endothelial cells and herpes-virus-1-infected peripheral blood mononuclear cells on human dermal microvascular endothelial cells was studied as a model of herpes-associated erythema multiforme. After infection of human dermal microvascular endothelial cells with native herpes virus and overnight culture, 60%--90% of human dermal microvascular endothelial cells showed cytopathic effects. HLA class I molecules and CD31 (PECAM-1) surface expression in herpes-virus-infected endothelial cells were substantially downregulated, whereas CD54 (ICAM-1) remained unchanged. Cocultivation with herpes-virus-1-infected peripheral blood mononuclear cells left characteristic plaques on the human dermal microvascular endothelial cell monolayer; however, very few human dermal microvascular endothelial cells (1%--3%) were infected. Adhesion molecule expression of human dermal microvascular endothelial cells cocultivated with herpes-virus-infected peripheral blood mononuclear cells demonstrated a 5-fold increase in CD54 expression, a 2-fold increase in HLA class I expression, but no change of CD31 by fluorescence-activated cell sorter analysis. Incubation of human dermal microvascular endothelial cells with ultraviolet-C irradiated herpes-virus-infected peripheral blood mononuclear cells had no effect on morphology or adhesion molecule expression levels. Changes of adhesion molecule expression by direct infection or cocultivation with peripheral blood mononuclear cells (with native and ultraviolet-C inactivated herpes virus infection) were also documented at the mRNA level. Adhesion assays demonstrated an increased binding of herpes-virus-infected peripheral blood mononuclear cells versus noninfected peripheral blood mononuclear cells to noninfected human dermal microvascular endothelial cells. Our results suggest that incubation of herpes-virus-infected peripheral blood mononuclear cells with human dermal microvascular endothelial cells induces significant upregulation of CD54 and major histocompatibility complex class I molecules in the surrounding noninfected human dermal microvascular endothelial cells, which is associated with an increased binding of peripheral blood mononuclear cells. Our in vitro findings may serve as a model for herpes-associated erythema multiforme possibly explaining the dermal inflammatory reaction seen in that condition.


Subject(s)
Endothelium, Vascular/cytology , Herpes Simplex/blood , Herpesvirus 1, Human , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/virology , Skin/cytology , Antigens, Surface/biosynthesis , Blotting, Northern , Cell Adhesion/physiology , Cell Communication , Cells, Cultured , Erythema Multiforme/virology , Histocompatibility Antigens Class I/biosynthesis , Humans , Intercellular Adhesion Molecule-1/biosynthesis , Microcirculation/cytology , Platelet Endothelial Cell Adhesion Molecule-1/biosynthesis
16.
Dtsch Med Wochenschr ; 125(15): 445-51, 2000 Apr 14.
Article in German | MEDLINE | ID: mdl-10800441

ABSTRACT

OBJECTIVE: Cytomegalovirus (CMV) infection is the most common viral infection in the early period after heart transplantation (HTX) and causes a significant morbidity and mortality. Although controversial, CMV is related to acute and chronic allograft rejection and to the development of graft vascular disease. It therefore plays an important role in the long-time outcome after solid organ transplantation. PATIENTS AND METHODS: 45 patients received a new heart between 1.1.97 and 31.12.1998. All of them were enrolled postoperatively in three-month antiviral prophylaxis (Cymevene). Only those patients were excluded from prophylaxis who were seronegative for CMV and received hearts from seronegative donors (n = 6). The pp65 antigenaemia assay and the murex hybrid capture CMV DNA assay on peripheral blood as well as the early antigen detection in the urine were used for CMV detection and also for monitoring. RESULTS: A total number of 580 assays were analysed (12.9 assays/patient). 561 tests (96.7%) were negative, 19 (3.3%) were positive. For CMV testing the pp65 antigenemia assay was used in 64.1%, the murex hybrid capture CMV DNA assay in 18.4% and the urine early antigen detection in 17.4%. Three patients (6.7%) developed viraemia during the first 3 postoperative months. Two patients (4.4%) suffered from CMV infection 8 and 9 months after heart transplantation and had to be treated with antiviral agents. Three patients (6.7%) died early after transplantation, but none had a CMV infection. CONCLUSION: Prevention of CMV disease was successful with three months of antiviral CMV prophylaxis after HTX. Asymptomatic viraemia during the prophylaxis period did not lead to tissue invasive disease. It is possible to carry out rapid CMV detection and CMV monitoring with the commercially available antigenaemia assays.


Subject(s)
Antiviral Agents/therapeutic use , Cytomegalovirus Infections/prevention & control , Ganciclovir/therapeutic use , Heart Transplantation , Postoperative Complications/prevention & control , Adolescent , Adult , Aged , Antigens, Viral/blood , Antigens, Viral/urine , Cytomegalovirus/genetics , Cytomegalovirus/immunology , Cytomegalovirus/isolation & purification , Cytomegalovirus Infections/complications , Cytomegalovirus Infections/diagnosis , DNA, Viral/analysis , Female , Graft Occlusion, Vascular/etiology , Graft Rejection/etiology , Humans , Male , Middle Aged , Retrospective Studies , Tissue Donors/statistics & numerical data , Viremia/prevention & control
17.
Am J Surg Pathol ; 24(4): 614-21, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10757411

ABSTRACT

Epstein-Barr virus (EBV)-associated smooth muscle tumors following solid organ transplantation are extremely rare, with only 12 cases reported in the literature thus far. The exact pathogenetic role of EBV infection in the oncogenesis of these soft tissue tumors in immunodeficient patients and the biologic behavior of such tumors is still unclear. We report a 26-year-old man in whom multiple smooth muscle tumors developed 36 to 51 months after heart transplantation. All tumors, two synchronous liver nodules, two subsequently occurring paravertebral tumors, and a single tumor in a vein at the left ankle were surgically resected. The tumor tissue was processed for routine histology and immunohistochemical (IHC) stains. Additionally, competitive polymerase-chain-reaction (PCR), reverse-transcriptase PCR (RT-PCR), as well as in situ hybridization (ISH) were used for EBV particle quantification and gene transcription analysis. The histologic features and immunohistochemical profiles were consistent with leiomyosarcoma in all tumor nodules. EBV infection was detected in >95% of tumor cell nuclei by EBER 1/2 ISH. Competitive PCR revealed 3105 EBV particles per milligram of tumor tissue. The EBV gene expression pattern analyzed by RT-PCR and IHC corresponded to the latency type III with specific expression of EBNA1, EBNA2, LMP1, and LMP2A genes. Under continuous antiviral therapy (famcyclovir) the patient currently shows no evidence of disease. Our data indicate that EBV infection plays a causal role in the development of smooth muscle tumors following organ transplantation. A latency type III, identical to EBV-associated posttransplant lymphoproliferative disorders, was identified and suggests a common pathogenetic mechanism in the development of these histogenetically distinct neoplasms. The fact that the patient currently shows no evidence of disease may be the result of the continuous administration of antiviral therapy because the soft tissue recurrences of the leiomyosarcoma occurred while the patient was not receiving antiviral prophylaxis.


Subject(s)
Heart Transplantation/adverse effects , Herpesviridae Infections/etiology , Herpesvirus 4, Human/isolation & purification , Leiomyosarcoma/etiology , Soft Tissue Neoplasms/etiology , Tumor Virus Infections/etiology , Adult , DNA, Viral/analysis , Herpesviridae Infections/pathology , Herpesviridae Infections/virology , Herpesvirus 4, Human/classification , Herpesvirus 4, Human/genetics , Humans , In Situ Hybridization , Leiomyosarcoma/pathology , Leiomyosarcoma/virology , Male , RNA, Messenger/analysis , RNA, Messenger/genetics , RNA, Viral/analysis , Receptors, Complement 3d/analysis , Reverse Transcriptase Polymerase Chain Reaction , Soft Tissue Neoplasms/pathology , Soft Tissue Neoplasms/virology , Tumor Virus Infections/pathology , Tumor Virus Infections/virology
18.
J Virol ; 73(9): 7722-33, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10438862

ABSTRACT

Epstein-Barr virus (EBV) is implicated in different central nervous system syndromes. The major cellular receptor for EBV, complement receptor type 2 (CR2) (CD21), is expressed by different astrocyte cell lines and human fetal astrocytes, suggesting their susceptibility to EBV infection. We demonstrated the infection of two astrocyte cell lines, T98 and CB193, at low levels. As infection was mediated by CR2, we used two stable CR2 transfectant astrocyte cell lines (T98CR2 and CB193CR2) to achieve a more efficient infection. We have monitored EBV gene expression for 2 months and observed the transient infection of T98 and T98CR2 cells and persistent infection of CB193 and CB193CR2 cells. The detection of BZLF1, BALF2, and BcLF1 mRNA expression suggests that the lytic cycle is initiated at early time points postinfection. At later time points the pattern of mRNA expressed (EBER1, EBNA1, EBNA2, and LMP1) differs from latency type III in the absence of LMP2A transcription and in the expression of BALF2 and BcLF1 but not BZLF1. A reactivation of the lytic cycle was achieved in CB193CR2 cells by the addition of phorbol esters. These studies identify astrocyte cell lines as targets for EBV infection and suggest that this infection might play a role in the pathology of EBV in the brain.


Subject(s)
Astrocytes/virology , Herpesvirus 4, Human/pathogenicity , Viral Proteins , Astrocytes/metabolism , DNA, Viral , DNA-Binding Proteins/genetics , Gene Expression , Genes, Viral , Genetic Vectors , Herpesvirus 4, Human/metabolism , Humans , RNA, Messenger , Receptors, Complement 3d/genetics , Receptors, Complement 3d/metabolism , Receptors, Virus/genetics , Receptors, Virus/metabolism , Trans-Activators/genetics , Transcription, Genetic , Transfection , Tumor Cells, Cultured
19.
J Immunol ; 161(9): 4604-10, 1998 Nov 01.
Article in English | MEDLINE | ID: mdl-9794388

ABSTRACT

To allow for a better characterization of the ligand binding structures of human complement receptor type 2 (CR2; CD21), we have established an IgG1 kappa mouse mAb, FE8, that interferes efficiently with binding of C3dg and EBV to CR2. In contrast to mAb OKB7, the only well-characterized mAb with similar specificity, mAb FE8 blocked binding of soluble C3dg or particles carrying multiple copies of surface-bound C3dg to CR2 or induced complete removal of these ligands from the receptor. In vitro EBV infection of B lymphocytes, on the other hand, was abrogated by mAbs FE8 and OKB7 with similar dose-response characteristics. As FE8 was shown to recognize a discontinuous epitope, a series of overlapping peptides derived from SCR1 and -2 and immobilized on cellulose was screened with FE8. The results suggest that up to five discontinuous sequences contributed to the epitope. The sequence 63-EYFNKYS-69, located between the two SCR units, reacted most intensively. Two other sequences, 16-YYSTPI-21 and 105-NGNKSVWCQANN-116, are located between Cys1 and Cys2 of SCR1 and around Cys3 of SCR2, respectively. Based on the solution structure for two factor H SCRs, a three-dimensional model of SCR1 and -2 was generated. The FE8 binding peptide sequences were located in relative proximity to each other, bounding the recess formed between SCR1 and -2. This potential of mAb FE8 is currently unique and may be exploited for interfering with conditions of unwanted recognition of C3dg-coated structures by the immune system.


Subject(s)
Complement C3b/metabolism , Peptide Fragments/metabolism , Protein Conformation , Receptors, Complement 3d/metabolism , Amino Acid Sequence , Animals , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/pharmacology , Antigen-Antibody Reactions , Binding Sites , Binding, Competitive , Computer Simulation , Consensus Sequence , Epitopes/immunology , Female , Herpesvirus 4, Human/metabolism , Humans , Hybridomas/immunology , Mice , Mice, Inbred BALB C , Microspheres , Models, Immunological , Models, Molecular , Molecular Sequence Data , Peptide Library , Protein Binding , Receptors, Complement 3d/chemistry , Receptors, Complement 3d/immunology , Repetitive Sequences, Amino Acid , Solubility
20.
Antiviral Res ; 38(1): 25-30, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9614001

ABSTRACT

N-chlorotaurine, an essential weak oxidant produced by stimulated human leukocytes, is known to have bactericidal, fungicidal and vermicidal properties. This study for the first time demonstrates its virucidal activity. By viral suspension tests at incubation times between 5 and 60 min, virus titers of both Herpes simplex virus type 1 and 2 were reduced about 1.3-2.9 log10 and 2.8-4.2 log10 by 0.1 and 1%, (5.5 and 55 mM) N-chlorotaurine, respectively. Virus titer reduction of adenovirus type 5 between 15 and 60 min was 0.5-2.0 and 0.6-4.0 log10, respectively, by the same concentrations of N-chlorotaurine. These findings support a contribution of N-chlorotaurine in destruction of pathogens during inflammatory reactions and also the possibility of its application as an antiviral agent in human medicine.


Subject(s)
Adenoviruses, Human/drug effects , Antiviral Agents/pharmacology , Herpesvirus 1, Human/drug effects , Herpesvirus 2, Human/drug effects , Taurine/analogs & derivatives , Adenoviruses, Human/growth & development , Animals , Cell Line , Chlorocebus aethiops , HeLa Cells , Herpesvirus 1, Human/growth & development , Herpesvirus 2, Human/growth & development , Humans , Taurine/pharmacology , Vero Cells
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