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1.
Antimicrob Agents Chemother ; 56(7): 3819-25, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22564837

RESUMEN

One recent, double-blind, randomized clinical trial with 200 patients showed that clarithromycin administered intravenously for 3 days in patients with ventilator-associated pneumonia (VAP) accelerated the resolution of pneumonia and decreased the risk of death from septic shock and multiple organ dysfunctions (MODS). The present study focused on the effect of clarithromycin on markers of inflammation in these patients. Blood was drawn immediately before the administration of the allocated treatment and on six consecutive days after the start of treatment. The concentrations of circulating markers were measured. Monocytes and neutrophils were isolated for immunophenotyping analysis and for cytokine stimulation. The ratio of serum interleukin-10 (IL-10) to serum tumor necrosis factor alpha (TNF-α) was decreased in the clarithromycin group compared with the results in the placebo group. Apoptosis of monocytes was significantly increased on day 4 in the clarithromycin group compared with the rate of apoptosis in the placebo group. On the same day, the expression of CD86 was increased and the ratio of soluble CD40 ligand (sCD40L) to CD86 in serum was unchanged. The release of TNF-α, IL-6, and soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) by circulating monocytes after stimulation was greater in the clarithromycin group than in the placebo group. The expression of TREM-1 on monocytes was also increased in the former group. These effects were pronounced in patients with septic shock and MODS. These results suggest that the administration of clarithromycin restored the balance between proinflammatory versus anti-inflammatory mediators in patients with sepsis; this was accompanied by more efficient antigen presentation and increased apoptosis. These effects render new perspectives for the immunotherapy of sepsis.


Asunto(s)
Claritromicina/uso terapéutico , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/patogenicidad , Neumonía Asociada al Ventilador/sangre , Neumonía Asociada al Ventilador/tratamiento farmacológico , Sepsis/sangre , Sepsis/tratamiento farmacológico , Apoptosis/efectos de los fármacos , Antígeno B7-2/sangre , Ligando de CD40/sangre , Método Doble Ciego , Humanos , Interleucina-10/sangre , Interleucina-6/sangre , Factor de Necrosis Tumoral alfa/sangre
2.
Crit Care ; 14(3): R96, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-20504311

RESUMEN

INTRODUCTION: Although major changes of the immune system have been described in sepsis, it has never been studied whether these may differ in relation to the type of underlying infection or not. This was studied for the first time. METHODS: The statuses of the innate and adaptive immune systems were prospectively compared in 505 patients. Whole blood was sampled within less than 24 hours of advent of sepsis; white blood cells were stained with monoclonal antibodies and analyzed though a flow cytometer. RESULTS: Expression of HLA-DR was significantly decreased among patients with severe sepsis/shock due to acute pyelonephritis and intraabdominal infections compared with sepsis. The rate of apoptosis of natural killer (NK) cells differed significantly among patients with severe sepsis/shock due to ventilator-associated pneumonia (VAP) and hospital-acquired pneumonia (HAP) compared with sepsis. The rate of apoptosis of NKT cells differed significantly among patients with severe sepsis/shock due to acute pyelonephritis, primary bacteremia and VAP/HAP compared with sepsis. Regarding adaptive immunity, absolute counts of CD4-lymphocytes were significantly decreased among patients with severe sepsis/shock due to community-acquired pneumonia (CAP) and intraabdominal infections compared with sepsis. Absolute counts of B-lymphocytes were significantly decreased among patients with severe sepsis/shock due to CAP compared with sepsis. CONCLUSIONS: Major differences of the early statuses of the innate and adaptive immune systems exist between sepsis and severe sepsis/shock in relation to the underlying type of infection. These results may have a major impact on therapeutics.


Asunto(s)
Inmunidad Adaptativa/inmunología , Inmunidad Innata/inmunología , Sepsis/clasificación , Anciano , Anciano de 80 o más Años , Apoptosis/inmunología , Linfocitos B/inmunología , Recuento de Linfocito CD4 , Femenino , Grecia , Antígenos HLA-DR/sangre , Humanos , Células Asesinas Naturales/inmunología , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Sepsis/sangre , Sepsis/inmunología
3.
Crit Care ; 13(6): R172, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19883512

RESUMEN

INTRODUCTION: The present study aimed to investigate changes of the immune response between sepsis due to ventilator-associated pneumonia (VAP) and sepsis due to other types of infections. METHODS: Peripheral venous blood was sampled from 68 patients with sepsis within 24 hours of diagnosis; 36 suffered from VAP; 32 from other nosocomial infections, all well-matched for severity, age and sex. Blood monocytes were isolated and cultured with/without purified endotoxin (lipopolysaccharide (LPS)). Estimation of tumour necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) in cultures' supernatants was done by an enzyme immunoassay. Flow cytometry was used to determine subpopulations of mononuclear cells and apoptosis. To mimic pathogenesis of VAP, mononuclear cells of healthy volunteers were progressively stimulated with increased inocula of pathogens; apoptosis was determined. RESULTS: In patients with VAP, the absolute number of CD3(+)/CD4(+) lymphocytes was significantly lower (P = 0.034) and apoptosis of isolated monocytes was increased (P = 0.007) compared to other infections. TNFalpha and IL-6 production from LPS-stimulated monocytes was lower in patients with VAP-related sepsis than with sepsis due to other infections. Apoptosis of monocytes was induced after in vitro stimulation of mononuclear cells by a mechanism mimicking VAP. CONCLUSIONS: Decrease of CD4-lymphocytes and immunoparalysis of monocytes are characteristic alterations of sepsis arising in the field of VAP.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Receptores de Lipopolisacáridos/inmunología , Monocitos/inmunología , Neumonía Asociada al Ventilador/inmunología , Sepsis/inmunología , Adolescente , Adulto , Anciano , Antígenos CD/inmunología , Apoptosis , Infecciones Bacterianas/complicaciones , Infecciones Bacterianas/inmunología , Recuento de Linfocito CD4 , Infección Hospitalaria/complicaciones , Infección Hospitalaria/inmunología , Femenino , Citometría de Flujo , Humanos , Lipopolisacáridos/farmacología , Masculino , Persona de Mediana Edad , Monocitos/efectos de los fármacos , Neumonía Asociada al Ventilador/complicaciones , Sepsis/sangre , Sepsis/etiología
4.
Diagn Microbiol Infect Dis ; 57(4): 419-22, 2007 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-17188449

RESUMEN

Infections by multidrug-resistant Acinetobacter baumannii constitute an increasing threat for critically ill patients. Colistin is often the only antimicrobial retaining activity against these strains. The postantibiotic effect (PAE) of colistin was studied on 19 isolates of A. baumannii resistant to ampicillin/sulbactam, ciprofloxacin, and carbapenems with the viable count method. The mean PAEs of 1x MIC and 4x MIC concentrations of colistin on the tested isolates were 3.90 and 4.48 h, respectively, indicating that a modified dosage scheme with increased dosing intervals might retain activity whereas minimizing the incidence of adverse effects.


Asunto(s)
Acinetobacter baumannii/efectos de los fármacos , Antibacterianos/farmacología , Colistina/farmacología , Farmacorresistencia Bacteriana Múltiple , Acinetobacter baumannii/crecimiento & desarrollo , Medios de Cultivo , Relación Dosis-Respuesta a Droga , Humanos , Pruebas de Sensibilidad Microbiana/normas
5.
Shock ; 26(4): 410-6, 2006 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-16980890

RESUMEN

Oleuropein, a novel immunomodulator derived from olive tree, was assessed in vitro and in experimental sepsis by Pseudomonas aeruginosa. After addition in monocyte and neutrophil cultures, malondialdehyde, TNF-alpha, IL-6, and bacterial counts were estimated in supernatants. Acute pyelonephritis was induced in 70 rabbits after inoculation of pathogen in the renal pelvis. Intravenous therapy was administered in four groups postchallenge by one multidrug-resistant isolate (A, controls; B, oleuropein; C, amikacin; D, both agents) and in three groups postchallenge by one susceptible isolate (E, controls; F, oleuropein; G, amikacin). Survival was recorded; bacterial growth in blood and organs was counted; endotoxins (LPS), malondialdehyde, total antioxidant status, and TNF-alpha in serum were estimated. TNF-alpha and IL-6 of cell supernatants were not increased compared with controls when triggered by LPS and P. aeruginosa. Counts of multidrug-resistant P. aeruginosa were decreased in monocyte supernatants. Median survival of groups A, B, C, D, E, F, and G were 3.00, 6.00, 2.00, 10.00, 1.00, 5.00, and 1.00 days, respectively. Bacteria in blood were lower at 48 h in groups B and D compared with A and in groups F and G compared with E. Total antioxidant status decreased steadily over time in groups A, C, D, and G, but not in groups B and F. TNF-alpha of groups B, C, and D was lower than A at 48 h. Tissue bacteria decreased in group F compared with E. Oleuropein prolonged survival in experimental sepsis probably by promoting phagocytosis or inhibiting biosynthesis of proinflammatory cytokines.


Asunto(s)
Factores Inmunológicos/farmacología , Infecciones por Pseudomonas/tratamiento farmacológico , Pseudomonas aeruginosa , Piranos/farmacología , Sepsis/tratamiento farmacológico , Sobrevida/psicología , Amicacina/farmacología , Animales , Antibacterianos/farmacología , Antioxidantes/farmacología , Células Cultivadas , Humanos , Glucósidos Iridoides , Iridoides , Masculino , Infecciones por Pseudomonas/mortalidad , Conejos , Sepsis/microbiología , Sepsis/mortalidad , Sobrevida/fisiología
6.
BMC Infect Dis ; 6: 142, 2006 Sep 18.
Artículo en Inglés | MEDLINE | ID: mdl-16981997

RESUMEN

BACKGROUND: Current theories underline the crucial role of pro-inflammatory mediators produced by monocytes for the pathogenesis of sepsis. Since monocytes derive from progenitor hemopoetic cells, the kinetics of stem cells was studied in peripheral blood of patients with sepsis. METHODS: Blood was sampled from 44 patients with septic syndrome due to ventilator-associated pneumonia on days 1, 3, 5 and 7 upon initiation of symptoms. Concentrations of tumour necrosis factor-alpha (TNFalpha), interleukin (IL)-6, IL-8 and G-CSF were estimated by ELISA. CD34/CD45 cells were determined after incubation with anti-CD45 FITC and anti-CD34 PE monocloncal antibodies and flow cytometric analysis. Samples from eight healthy volunteers served as controls. RESULTS: Median of CD34/CD45 absolute count of controls was 1.0/mul. Respective values of the total study population were 123.4, 112.4, 121.5 and 120.9/mul on days 1, 3, 5 and 7 (p < 0.0001 compared to controls). Positive correlations were found between the absolute CD34/CD45 count and the absolute monocyte count on days 1, 5 and 7. Survival was prolonged among patients with less than 310/microl CD34/CD45 cells on day 1 compared to those with more than 310/microl of CD34/CD45 cells (p: 0.022). Hazard ratio for death due to sepsis was 5.47 (p: 0.039) for CD34/CD45 cells more than 310/microl. Median IL-6 on day 1 was 56.78 and 233.85 pg/ml respectively for patients with less than 310/microl and more than 310/microl CD34/CD45 cells (p: 0.021). CONCLUSION: Stem cells are increased in peripheral blood over all days of follow-up compared to healthy volunteers. Patients with counts on day 1 less than 310/microl are accompanied by increased survival compared to patients with more than 310/microl.


Asunto(s)
Células Madre Hematopoyéticas/patología , Sepsis/patología , Adulto , Anciano , Citocinas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Neumonía/complicaciones , Sepsis/complicaciones , Choque Séptico/complicaciones , Choque Séptico/patología , Análisis de Supervivencia , Ventiladores Mecánicos/efectos adversos
7.
Crit Care ; 10(6): R166, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-17129388

RESUMEN

INTRODUCTION: Our aim was to define early changes of lymphocytes and of NK cells in severe sepsis and to correlate them with serum levels of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1). METHODS: Blood was sampled from 49 patients with proven highly suspected infection by Gram-negative pathogens, within 12 hours of the advent of severe sepsis, and was also sampled from six healthy volunteers. White blood cells were targeted with monoclonal antibodies and were analyzed by flow cytometry. The concentrations of sTREM-1 were estimated by ELISA. RESULTS: The presence of CD3/CD4 cells was significantly lower (P < 0.0001) and that of NK cells significantly higher among patients with sepsis compared with controls (P = 0.011). The proportions (median +/- standard error) of ANNEXIN-V/CD4/CD3-positive cells, of ANNEXIN-V/CD8/CD3-positive cells and of ANNEXIN-V/CD14-positive cells of the patient population were 7.41 +/- 2.26%, 7.69 +/- 3.42% and 1.96 +/- 4.22%, respectively. Patients with NK cells >20% survived longer compared with those patients with NK cells < or =20% (P = 0.041), and patients with sTREM-1 concentrations >180 pg/ml survived longer compared with those patients with sTREM-1 concentrations < or =180 pg/ml (P = 0.042). A negative correlation was found between the percentages of ANNEXIN-V/CD4/CD3-positive cells and of CD3/CD4 cells (rs = -0.305, P = 0.049), and a positive correlation was found between the serum sTREM-1 concentration and the percentage of NK cells (rs = +0.395, P = 0.014). NK cells isolated from two healthy volunteers released sTREM-1 upon triggering with endotoxins. CONCLUSION: Early severe sepsis is characterized by CD4-lymphopenia and increased NK cells, providing a survival benefit for the septic patient at percentages >20%. The survival benefit resulting from elevated NK cells might be connected to elevated serum levels of sTREM-1.


Asunto(s)
Linfocitos T CD4-Positivos , Infecciones por Bacterias Gramnegativas/inmunología , Células Asesinas Naturales , Sepsis/inmunología , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Infecciones por Bacterias Gramnegativas/sangre , Humanos , Recuento de Linfocitos , Masculino , Glicoproteínas de Membrana/sangre , Pronóstico , Receptores Inmunológicos/sangre , Sepsis/sangre , Sepsis/microbiología , Análisis de Supervivencia , Receptor Activador Expresado en Células Mieloides 1
8.
World J Gastroenterol ; 12(41): 6711-4, 2006 Nov 07.
Artículo en Inglés | MEDLINE | ID: mdl-17075990

RESUMEN

AIM: To unravel the differences between systematic inflammatory response syndrome (SIRS) of acute pancreatitis compared to the same syndrome in sepsis. METHODS: Twenty-five patients were enrolled, 12 with sepsis and 13 acute pancreatitis. After diagnosis 20 mL blood was sampled. Half were assayed for isolation of monocytes and 10 mL was centrifuged for serum test of tumor necrosis factor alpha (TNFalpha and interleukin-6 (IL-6). Half of monocytes were incubated in the presence of patients' serum and supernatants were collected. The other half was treated for estimation of optical photometry under caspase-3 inhibition. TNFalpha and IL-6 were estimated by an enzyme immunoassay. RESULTS: median+/-SE of serum IL-6 in septic patients and acute pancreatitis patients was 192.30+/-35.40 ng/L and 21.00+/-16.05 ng/L, respectively (P<0.01). Respective values of caspase-3 were 0.94+/-0.17 pmol/min 10(4) cells and 0.34+/-0.09 pmol/min 10(4) cells (P<0.05). IL-6 of monocyte supernatants of patients with sepsis was significantly increased after addition of patients' serum, while that of patients with acute pancreatitis did not show significant difference. CONCLUSION: The data have shown that monocyte activity is different between acute pancreatitis and sepsis. This phenomenon might be explained as a different pathway to the pro-inflammatory cytokines release or could be a novel anti-inflammatory response in acute pancreatitis.


Asunto(s)
Monocitos/patología , Pancreatitis/sangre , Sepsis/sangre , Síndrome de Respuesta Inflamatoria Sistémica/sangre , Enfermedad Aguda , Adulto , Anciano , Caspasa 3/sangre , Diagnóstico Diferencial , Humanos , Interleucina-6/sangre , Persona de Mediana Edad , Monocitos/metabolismo , Pancreatitis/complicaciones , Pancreatitis/diagnóstico , Sepsis/complicaciones , Sepsis/diagnóstico , Síndrome de Respuesta Inflamatoria Sistémica/diagnóstico , Síndrome de Respuesta Inflamatoria Sistémica/etiología , Factor de Necrosis Tumoral alfa/sangre
9.
Pediatr Dent ; 26(1): 23-8, 2004.
Artículo en Inglés | MEDLINE | ID: mdl-15080354

RESUMEN

PURPOSE: The purpose of this controlled clinical trial was to compare the working time for caries removal in primary teeth, the need for local anesthesia and patient cooperation, when the chemomechanical Carisolv or the conventional mechanical method were used. METHODS: The sample consisted of primary teeth of children who had occlusal or buccal carious lesions into dentin. High speed and/or low speed were used as the conventional mechanical method of caries removal. The efficiency in caries removal was judged on the basis of clinical criteria. Length of working time, need of local anesthesia, and level of patient cooperation were recorded for both methods. Statistical analysis was performed using the student's t test and chi-square test. RESULTS: Working time with the chemomechanical method was much more prolonged than with the mechanical method (P < .001), but it did not negatively affect children's cooperation. Furthermore, the chemomechanical method reduced the need for administration of local anesthesia for Class V cavity preparations. CONCLUSIONS: The chemomechanical method, although more prolonged, is effective in caries removal in primary teeth, does not influence children's cooperation and may reduce the need of local anesthesia in Class V restorations.


Asunto(s)
Caries Dental/terapia , Preparación de la Cavidad Dental/métodos , Ácido Glutámico/uso terapéutico , Leucina/uso terapéutico , Lisina/uso terapéutico , Diente Primario/patología , Anestesia Dental , Anestesia Local , Distribución de Chi-Cuadrado , Niño , Preescolar , Conducta Cooperativa , Preparación de la Cavidad Dental/instrumentación , Equipo Dental de Alta Velocidad , Restauración Dental Permanente , Dentina/patología , Femenino , Humanos , Masculino , Estudios Prospectivos , Factores de Tiempo
10.
APMIS ; 122(1): 68-75, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23656439

RESUMEN

Evidence from a recent randomized study of our group suggests that intravenous clarithromycin resulted in earlier resolution of ventilator-associated pneumonia. The need to understand the mechanism of action of clarithromycin guided to the study of a model of experimental empyema by multidrug-resistant Pseudomonas aeruginosa in 40 rabbits. Animals were randomized into controls (group A); treatment with clarithromycin (group B); treatment with piperacillin/tazobactam (group C); and treatment with both agents (group D). Pleural fluid was collected at regular time intervals for quantitative culture, estimation of cell apoptosis and of concentrations of tumour necrosis factor-alpha (TNFα). After 7 days, animals were euthanized for estimation of tissue growth. Bacterial growth in the pleural fluid of group D was significantly decreased compared with the other groups on day 5. Lung growth of group D was lower than group A. That was also the case of cytokine stimulation by pleural fluid samples on U937 monocytes. It is concluded that administration of clarithromycin enhanced the antimicrobial efficacy of piperacillin/tazobactam and decreased bacterial growth in the pleural fluid and in tissues. It also attenuated the pro-inflammatory phenomena induced by the ß-lactam.


Asunto(s)
Antibacterianos/uso terapéutico , Claritromicina/uso terapéutico , Empiema Pleural/tratamiento farmacológico , Empiema Pleural/microbiología , Infecciones por Pseudomonas/tratamiento farmacológico , Infecciones por Pseudomonas/microbiología , Pseudomonas aeruginosa , Adyuvantes Inmunológicos/administración & dosificación , Adyuvantes Inmunológicos/uso terapéutico , Animales , Antibacterianos/administración & dosificación , Apoptosis/efectos de los fármacos , Apoptosis/inmunología , Claritromicina/administración & dosificación , Modelos Animales de Enfermedad , Farmacorresistencia Bacteriana Múltiple , Quimioterapia Combinada , Empiema Pleural/inmunología , Humanos , Interleucina-6/biosíntesis , Masculino , Ácido Penicilánico/administración & dosificación , Ácido Penicilánico/análogos & derivados , Piperacilina/administración & dosificación , Combinación Piperacilina y Tazobactam , Derrame Pleural/inmunología , Derrame Pleural/microbiología , Infecciones por Pseudomonas/inmunología , Pseudomonas aeruginosa/efectos de los fármacos , Conejos , Factor de Necrosis Tumoral alfa/biosíntesis , Células U937
11.
J Leukoc Biol ; 89(3): 343-9, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21030664

RESUMEN

We hypothesized that a factor may circulate in serum early during sepsis, modulating apoptosis of monocytes and lymphocytes. Serum was collected from 20 healthy volunteers and from 48 patients with severe sepsis/shock within 12 h from signs of the first failing organ. PBMCs were isolated from 20 healthy volunteers and incubated with collected sera. Apoptosis and expression of CD95 were determined by flow cytometry; experiments were run in the presence of caspase-8 and caspase-9 inhibitors and of CaCl(2). Activity of caspase-3 was determined in cell lysates by a chromogenic kinetic assay. Incubation with serum of patients induced apoptosis of CD4 lymphocytes and inhibited apoptosis of CD14 monocytes. This was attenuated after diluting serum or mixing with healthy serum. Activity of caspase-3 was consistent with these findings. Induced apoptosis of CD4 lymphocytes was greater among nonsurvivors, and it was inhibited in the presence of caspase inhibitors. Inhibitors did not modify the effect of patients' serum on apoptosis of CD14 monocytes. CaCl(2) reversed the inhibitory effect on apoptosis of CD14 moncytes. The above findings support the hypothesis for the existence of an early circulating factor in severe sepsis/shock, modulating apoptosis of CD4 lymphocytes and of CD14 monocytes by interaction with the two apoptotic pathways.


Asunto(s)
Apoptosis , Linfocitos T CD4-Positivos/patología , Factores Inmunológicos/sangre , Monocitos/patología , Sepsis/sangre , Anciano , Linfocitos T CD4-Positivos/enzimología , Caspasa 3/metabolismo , Extractos Celulares , Separación Celular , Demografía , Femenino , Humanos , Receptores de Lipopolisacáridos/metabolismo , Masculino , Monocitos/enzimología , Receptor fas/metabolismo
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