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1.
J Antimicrob Chemother ; 69(12): 3387-92, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25103492

RESUMO

BACKGROUND: Bloodstream infections (BSIs) caused by enterobacteria remain a leading cause of mortality in patients with chemotherapy-induced neutropenia. The rate and type of colonization and infection with ESBL-producing Enterobacteriaceae (ESBL-E) and their mode of transmission in German cancer centres is largely unknown. METHODS: We performed a prospective, observational study at five German university-based haematology departments. Participating sites screened for intestinal ESBL-E colonization within 72 h of admission, every 10 ± 2 days thereafter and before discharge. Three of the five centres performed contact isolation for patients colonized or infected with ESBL-E. Molecular characterization of resistance mechanisms and epidemiological typing of isolates by repetitive extragenic palindromic PCR (rep-PCR) and PFGE was performed to assess strain transmission between patients. RESULTS: Between October 2011 and December 2012, 719 hospitalizations of 497 haematological high-risk patients comprising 20,143 patient-days were analysed. Mean duration of in-hospital stay was 36.6 days (range: 2-159 days). ESBL-E were identified from screening samples (82.8% Escherichia coli and 14.6% Klebsiella pneumoniae) in 55/497 patients (11.1%; range by centre: 5.8%-23.1%). PFGE and rep-PCR revealed only a single case of potential cross-transmission among two patients colonized with K. pneumoniae. Six episodes of BSI with ESBL-E were observed. Multivariate analysis revealed previous colonization with ESBL-E as the most important risk factor for BSI with ESBL-E (OR 52.00; 95% CI 5.71-473.89). CONCLUSIONS: Even though BSI with ESBL-E is still rare in this high-risk population, colonization rates are substantial and vary considerably between centres. In-hospital transmission of ESBL-E as assessed by molecular typing was the exception.


Assuntos
Portador Sadio/epidemiologia , Portador Sadio/microbiologia , Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/enzimologia , Neoplasias Hematológicas/complicações , beta-Lactamases/metabolismo , Adolescente , Adulto , Idoso , Bacteriemia/epidemiologia , Bacteriemia/microbiologia , Estudos de Coortes , Eletroforese em Gel de Campo Pulsado , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Feminino , Genótipo , Alemanha/epidemiologia , Hospitais Universitários , Humanos , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Reação em Cadeia da Polimerase , Estudos Prospectivos , Adulto Jovem , beta-Lactamases/classificação
2.
In Vivo ; 21(1): 17-23, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17354609

RESUMO

Viral and plasmid vectors may cause immunological side-effects resulting from the expression of therapeutically unwanted genes and from CpG motifs contained in their sequence. A new vector type for minimalistic, immunological-defined gene expression (MIDGE) may overcome these problems. MIDGE is a minimal size gene transfer unit consisting of the expression cassette, including promotor, gene and RNA-stabilizing sequences, flanked by two short hairpin oligonucleotide sequences. DNA not encoding the desired gene is reduced to a minimum. To compare transfection efficiencies in vivo hydrodynamics-based, systemic transfection was performed in BALB/c mice with MIDGE vectors and corresponding plasmids. The transfection efficiencies of the MIDGE vectors as measured by luciferase expression were significantly higher in liver (2.5-fold), lung (3.5-fold), kidneys (3.9-fold) and heart (17-fold) as compared to plasmids. The mean numbers of MIDGE vector molecules per cell as measured by quantitative PCR were also significantly higher. These advantages suggest the preferential use of this new vector type for clinical gene therapy studies.


Assuntos
Ilhas de CpG , Vetores Genéticos , Transfecção/métodos , Transgenes , Animais , Luciferases/biossíntese , Luciferases/genética , Camundongos , Camundongos Endogâmicos BALB C , Especificidade de Órgãos , Plasmídeos , Regiões Promotoras Genéticas
3.
Immunol Cell Biol ; 83(3): 278-85, 2005 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15877606

RESUMO

Monocyte-derived dendritic cells (mDC), the most frequently applied DC subset in clinical studies, which can be obtained easily from peripheral blood monocytes after incubation with GM-CSF and IL-4, have not been clearly demonstrated to be activated by CpG oligodeoxynucleotides (ODN). The development of novel molecular strategies - such as the use of CpG-ODN - to increase immunological functions and thus improve the therapeutic efficiency of mDC vaccines in the treatment of malignant diseases is highly desirable. CpG-ODN need to be internalized into specific intracellular compartments to be active. Therefore, we applied electroporation and lipofection and compared these techniques with incubation to overcome possible defects in localization. Conditions of CpG-ODN transfection of these cells were optimized using fluorescein-marked ODN 2216. We were able to achieve high transfection efficiencies with various methods of delivery. However, we did not observe increased expression of maturation-associated and functionally relevant surface antigens (CD14, HLA-DR, CD40, CD83, CD80 and CD86), significant secretion of IL-12 and IFN-alpha in culture supernatant, or enhanced antitumour activation of cytokine-induced killer cells. In conclusion, our results show that non-viral transfection of CpG-ODN is not sufficient to overcome resistance of mDC to CpG activation.


Assuntos
Células Dendríticas/imunologia , Monócitos/citologia , Oligodesoxirribonucleotídeos/imunologia , Transfecção/métodos , Antígenos CD/metabolismo , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Técnicas de Cocultura , Citotoxicidade Imunológica/imunologia , Células Dendríticas/metabolismo , Eletroporação , Ácidos Graxos Monoinsaturados/química , Citometria de Fluxo , Substâncias de Crescimento/farmacologia , Antígenos HLA-DR/metabolismo , Humanos , Interferon-alfa/metabolismo , Interleucina-12/metabolismo , Células Matadoras Naturais/imunologia , Lipopolissacarídeos/farmacologia , Oligodesoxirribonucleotídeos/genética , Compostos de Amônio Quaternário/química
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