RESUMEN
BACKGROUND: To demonstrate whether extensive intraoperative peritoneal lavage (EIPL) could yield better results in overall survival and less recurrence, regardless of peritoneal cytology, compared to standard peritoneal lavage (SPL). METHODS: A prospective randomised multicenter study including 94 patients (47 per arm) to detect a 20% difference in 3-year overall survival in patients with locally advanced tumours without peritoneal carcinomatosis. Three samples of peritoneal fluid were obtained (at the beginning, the end of procedure and after the assigned peritoneal lavage). Clinicopathological and surgical data were analysed by group. Postoperative complications, location of recurrence and surgical approach were evaluated. Overall survival was calculated by the Kaplan-Meier method and the uni/multivariate analysis for prognostic factors was carried out using Cox regression analysis. RESULTS: A total of 86 patients were analysed (4 excluded per group). No statistical differences were observed in clinicopathological or surgical data between groups, considering both groups well-balanced for analysis. Overall survival at 3 years was 64.3% for SPL vs. 62.3% for EIPL (p 0.421). Only three patients had at least one positive peritoneal cytology (1:2). There were no differences regarding postoperative complications (SPL: 37.2% vs. EIPL: 32.5%, p 0.65) or between location of recurrence and number of recurrences. The number of recurrences did not differ between surgical approaches, but locoregional and peritoneal recurrences were fewer with the laparoscopic approach (p 0.048). CONCLUSIONS: The regular use of extensive peritoneal lavage in patients with locally advanced gastric cancer, regardless of peritoneal cytology, has not been effective as prophylaxis of peritoneal recurrence or better survival.
Asunto(s)
Cuidados Intraoperatorios/mortalidad , Recurrencia Local de Neoplasia/mortalidad , Lavado Peritoneal/métodos , Neoplasias Peritoneales/mortalidad , Neoplasias Gástricas/mortalidad , Anciano , Análisis de Varianza , Quimioterapia Adyuvante , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Invasividad Neoplásica , Recurrencia Local de Neoplasia/tratamiento farmacológico , Recurrencia Local de Neoplasia/prevención & control , Recurrencia Local de Neoplasia/secundario , Lavado Peritoneal/mortalidad , Neoplasias Peritoneales/tratamiento farmacológico , Neoplasias Peritoneales/prevención & control , Neoplasias Peritoneales/secundario , Estudios Prospectivos , Neoplasias Gástricas/tratamiento farmacológico , Neoplasias Gástricas/patología , Neoplasias Gástricas/cirugíaRESUMEN
UNLABELLED: Two bacteriophages, isolated from faeces, were assayed as biocontrol agents of pathogenic Escherichia coli during milk fermentation. Phage DT1 was tested on the strain E. coli DH5α, one enteropathogenic E. coli (EPEC) strain and one Shiga toxigenic E. coli O157:H7 (STEC) strain. Phage DT6 was tested on two STEC strains (O157:H7 and non-O157). One additional assay was performed by using a cocktail of both phages against the O157:H7 STEC strain. Streptococcus thermophilus 10-C, the strain used as lactic starter, reached 10(9) CFU ml(-1) after 4 h, while pH values fell to 4·5 after 8 h, regardless of the presence of E. coli strains and/or phages. In absence of phages, E. coli strains reached 4-6 log CFU ml(-1) at 5-6 h. Escherichia coli DH5α and O157:H7 STEC strains were rapidly and completely inactivated by phage DT1 and phage cocktail, respectively, while O157:H7 STEC was completely inactivated either by DT1 or by DT6, after 8 h. The EPEC strain was not detected at 1 h (<10 CFU ml(-1) ) but grew afterwards, though at lower rates than without phage. For non-O157:H7 STEC, reductions lower than 1 log CFU ml(-1) were observed for all sampling times. Phages DT1 and DT6, either individually or as a cocktail, effectively reduce O157:H7 STEC counts during milk fermentation, without compromising the starter culture performance. SIGNIFICANCE AND IMPACT OF THE STUDY: Coliphages DT1 and DT6, isolated from faeces and selected on the basis of their host range, showed to be valuable tools for the control of pathogenic Escherichia coli during milk fermentation, without compromising the starter culture performance. Both phages, either individually or as a cocktail, may function as an extra safety barrier beyond traditional pasteurization, effectively reducing O157:H7 Shiga toxin-producing Escherichia coli (STEC) counts during early growth, thus avoiding Shiga toxin production and accumulation.
Asunto(s)
Agentes de Control Biológico , Colifagos , Escherichia coli O157/virología , Heces/virología , Leche/microbiología , Escherichia coli Shiga-Toxigénica/virología , Animales , Bovinos , Escherichia coli O157/crecimiento & desarrollo , Fermentación , Escherichia coli Shiga-Toxigénica/crecimiento & desarrollo , Streptococcus thermophilus/crecimiento & desarrolloRESUMEN
2,4-Dichlorophenoxyacetic acid (2,4-D) is a herbicide widely used in the world and mainly excreted by the renal route in exposed humans and animals. Herbicides can affect other nontarget organisms, such as Escherichia coli. We observed that a single exposure to 1 mM 2,4-D diminished growth and total protein content in all E. coli strains tested in vitro. In addition, successive exposures to 0.01 mM 2,4-D had a toxic effect decreasing growth up to early stationary phase. Uropathogenic E. coli adhere to epithelial cells mediated by fimbriae, adhesins, and hydrophobic properties. 2,4-D exposure of uropathogenic E. coli demonstrated altered hydrophobicity and fimbriation. Hydrophobicity index values obtained by partition in p-xylene/water were 300-420% higher in exposed cells than in control ones. Furthermore, values of hemagglutination titer, protein contents in fimbrial crude extract, and electron microscopy demonstrated a significant diminution of fimbriation in treated cells. Other envelope alterations could be detected, such as lipoperoxidation, evidenced by decreased polyunsaturated fatty acids and increased lipid degradation products (malonaldehyde), and motility diminution. These alterations decreased cell adherence to erythrocytes, indicating a diminished pathogenic capacity of the 2,4-D-exposed E. coli.
Asunto(s)
Ácido 2,4-Diclorofenoxiacético/toxicidad , Escherichia coli/efectos de los fármacos , Herbicidas/toxicidad , Ácido 2,4-Diclorofenoxiacético/farmacocinética , Adhesinas de Escherichia coli/efectos de los fármacos , Membrana Celular/efectos de los fármacos , Escherichia coli/química , Escherichia coli/crecimiento & desarrollo , Peroxidación de Lípido/efectos de los fármacosRESUMEN
Clofibric and ethacrynic acids are prototypical pharmacological agents administered in the treatment of hypertrigliceridemia and as a diuretic agent, respectively. They share with 2,4-dichlorophenoxyacetic acid (the widely used herbicide known as 2,4-D) a chlorinated phenoxy structural moiety. These aryloxoalcanoic agents (AOAs) are mainly excreted by the renal route as unaltered or conjugated active compounds. The relatedness of these agents at the structural level and their potential effect on therapeutically treated or occupationally exposed individuals who are simultaneously undergoing a bacterial urinary tract infection led us to analyze their action on uropathogenic, clinically isolated Escherichia coli strains. We found that exposure to these compounds increases the bacterial resistance to an ample variety of antibiotics in clinical isolates of both uropathogenic and nonpathogenic E. coli strains. We demonstrate that the AOAs induce an alteration of the bacterial outer membrane permeability properties by the repression of the major porin OmpF in a micF-dependent process. Furthermore, we establish that the antibiotic resistance phenotype is primarily due to the induction of the MarRAB regulatory system by the AOAs, while other regulatory pathways that also converge into micF modulation (OmpR/EnvZ, SoxRS, and Lrp) remained unaltered. The fact that AOAs give rise to uropathogenic strains with a diminished susceptibility to antimicrobials highlights the impact of frequently underestimated or ignored collateral effects of chemical agents.
Asunto(s)
Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Porinas/efectos de los fármacos , Infecciones Urinarias/microbiología , Resistencia a Múltiples Medicamentos , Escherichia coli/enzimología , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Porinas/genética , beta-Galactosidasa/metabolismoRESUMEN
An hemagglutination (HA) type system has been applied to demonstrate mannose sensitive (MS) and mannose resistant (MR) hemagglutination produced by Escherichia coli isolated from urinary tract infections. Hemagglutination types were obtained by the agglutination of different species of red cells -human, bovine, chicken and guinea pig- suspended in buffer phosphate (PBS), with and without mannose, with E. coli cells grown in CFA agar (Casamino acid 10 g, yeast extract 15 g, sodium chloride 2.5 g, potassium phosphate 8.7 g, magnesium sulfate 0.5 g, manganese chloride 0.005 g, agar 20 g). Salting out (hydrophobicity) and yeast agglutination assays were performed for a complete evaluation of results. The applicability of this system was based on the dates exhibited in Table 1. A significant proportion (45%) of the uropathogenic Escherichia coli strains showed RNNN HA patterns, and (16%) NNSS, and (15%) SNSS were also considered important. The application of this hemagglutination system on this kind of strains allowed the evaluation of the different types of hemagglutination and their relation with colonization capacity.
Asunto(s)
Escherichia coli/clasificación , Infecciones Urinarias/microbiología , Animales , Pruebas de Hemaglutinación , HumanosRESUMEN
La presencia de células con características especiales denominadas células centelleantes, en el sedimento urinario provenientes de infecciones urinarias producidas por cepas de enterobacterias que expresan fimbrias Pap, fue evaluada en este estudio. En 143 cepas de enterobacterias, la capacidad de adhesión fue analizada aglutinación de glóbulos rojos humanos grupo A suspendidos en buffer fosfato (PBS), para ver hemoaglutinación (HA), y con el agregado de D-manosa al 1 por ciento para detectar hemoaglutinación manosa-sensible (HAMS) o manosa resistente (HAMR). Escherichia coli presentó una diferencia significativa en el número de cepas que expresan fimbrias manosa resistente (MR):56 y las que expresan fimbrias manosa sensibles (MS):25. Las especies MR, fueron enfrentadas con glóbulos rojos humanos grupo P y eritrocitos de cerdo, utilizando como inhibidor de la hemoaglutinación el digalactósido (1-4) -ß-D galactopiranósido. No se estableció una diferencia notable en el número de cepas con fimbrias MR, que presentaron capacidad hemoaglutinante, frente a las dos especies de eritrocitos. Pero fue comparativamente mayor, el número de cepas con fimbrias Pap que aglutinaron glóbulos rojos humanos grupo P. La relación presencia de células centelleantes en el sedimento urinario y cepas de enterobacterias que expresan fimbrias MR, como agente etiológico del proceso infeccioso, fue significativamente mayor que la establecida por cepas con fimbrias MS (AU)
Asunto(s)
Humanos , Masculino , Femenino , Infecciones Urinarias/microbiología , Manosa/diagnóstico , Infecciones por Enterobacteriaceae/orina , Enterobacteriaceae/patogenicidad , Pruebas de Inhibición de Hemaglutinación/estadística & datos numéricos , Pruebas de Inhibición de Hemaglutinación/métodos , Orina/citología , Orina/microbiología , Infecciones por Enterobacteriaceae/diagnóstico , Infecciones Urinarias/diagnóstico , Adhesinas Bacterianas/efectos adversos , Fimbrias Bacterianas/química , Enterobacteriaceae/aislamiento & purificación , Enterobacteriaceae/clasificaciónRESUMEN
La presencia de células con características especiales denominadas células centelleantes, en el sedimento urinario provenientes de infecciones urinarias producidas por cepas de enterobacterias que expresan fimbrias Pap, fue evaluada en este estudio. En 143 cepas de enterobacterias, la capacidad de adhesión fue analizada aglutinación de glóbulos rojos humanos grupo A suspendidos en buffer fosfato (PBS), para ver hemoaglutinación (HA), y con el agregado de D-manosa al 1 por ciento para detectar hemoaglutinación manosa-sensible (HAMS) o manosa resistente (HAMR). Escherichia coli presentó una diferencia significativa en el número de cepas que expresan fimbrias manosa resistente (MR):56 y las que expresan fimbrias manosa sensibles (MS):25. Las especies MR, fueron enfrentadas con glóbulos rojos humanos grupo P y eritrocitos de cerdo, utilizando como inhibidor de la hemoaglutinación el digalactósido (1-4) -ß-D galactopiranósido. No se estableció una diferencia notable en el número de cepas con fimbrias MR, que presentaron capacidad hemoaglutinante, frente a las dos especies de eritrocitos. Pero fue comparativamente mayor, el número de cepas con fimbrias Pap que aglutinaron glóbulos rojos humanos grupo P. La relación presencia de células centelleantes en el sedimento urinario y cepas de enterobacterias que expresan fimbrias MR, como agente etiológico del proceso infeccioso, fue significativamente mayor que la establecida por cepas con fimbrias MS
Asunto(s)
Humanos , Masculino , Femenino , Infecciones por Enterobacteriaceae/orina , Enterobacteriaceae/patogenicidad , Manosa , Infecciones Urinarias/microbiología , Adhesinas Bacterianas/efectos adversos , Infecciones por Enterobacteriaceae/diagnóstico , Enterobacteriaceae/clasificación , Enterobacteriaceae/aislamiento & purificación , Fimbrias Bacterianas/química , Pruebas de Inhibición de Hemaglutinación/estadística & datos numéricos , Pruebas de Inhibición de Hemaglutinación/métodos , Infecciones Urinarias/diagnóstico , Orina/citología , Orina/microbiologíaRESUMEN
An hemagglutination (HA) type system has been applied to demonstrate mannose sensitive (MS) and mannose resistant (MR) hemagglutination produced by Escherichia coli isolated from urinary tract infections. Hemagglutination types were obtained by the agglutination of different species of red cells -human, bovine, chicken and guinea pig- suspended in buffer phosphate (PBS), with and without mannose, with E. coli cells grown in CFA agar (Casamino acid 10 g, yeast extract 15 g, sodium chloride 2.5 g, potassium phosphate 8.7 g, magnesium sulfate 0.5 g, manganese chloride 0.005 g, agar 20 g). Salting out (hydrophobicity) and yeast agglutination assays were performed for a complete evaluation of results. The applicability of this system was based on the dates exhibited in Table 1. A significant proportion (45
) of the uropathogenic Escherichia coli strains showed RNNN HA patterns, and (16
) NNSS, and (15
) SNSS were also considered important. The application of this hemagglutination system on this kind of strains allowed the evaluation of the different types of hemagglutination and their relation with colonization capacity.
RESUMEN
An hemagglutination (HA) type system has been applied to demonstrate mannose sensitive (MS) and mannose resistant (MR) hemagglutination produced by Escherichia coli isolated from urinary tract infections. Hemagglutination types were obtained by the agglutination of different species of red cells -human, bovine, chicken and guinea pig- suspended in buffer phosphate (PBS), with and without mannose, with E. coli cells grown in CFA agar (Casamino acid 10 g, yeast extract 15 g, sodium chloride 2.5 g, potassium phosphate 8.7 g, magnesium sulfate 0.5 g, manganese chloride 0.005 g, agar 20 g). Salting out (hydrophobicity) and yeast agglutination assays were performed for a complete evaluation of results. The applicability of this system was based on the dates exhibited in Table 1. A significant proportion (45
) of the uropathogenic Escherichia coli strains showed RNNN HA patterns, and (16
) NNSS, and (15
) SNSS were also considered important. The application of this hemagglutination system on this kind of strains allowed the evaluation of the different types of hemagglutination and their relation with colonization capacity.