Changing epidemiology of extended-spectrum ß-lactamases in Argentina: emergence of CTX-M-15.

A multicenter survey, carried out in 2010 in Argentina, showed an increased prevalence of extended-spectrum ß-lactamase (ESBL)-producing enterobacteria, with some changes in the molecular epidemiology of circulating ESBLs. While enzymes of the CTX-M-2 group remain endemic, the emergence of CTX-M-15 and of enzymes of the CTX-M-8 and CTX-M-9 groups was observed. The CTX-M-15-positive isolates represented 40% of CTX-M producers and included representatives of Escherichia coli ST131 and Klebsiella pneumoniae ST11.

Caracterización fenotípica y genotípica de la resistencia a imipenem en Pseudomonas aeruginosa aisladas en un hospital de Buenos Aires / Phenotypic and genotypic characterization of imipenem-resistant Pseudomonas aeruginosa isolated in Buenos Aires

En el presente estudio, que tuvo por objeto analizar los mecanismos involucrados en la resistencia a carbapenemes, se incluyeron 129 aislamientos de Pseudomonas aeruginosa recuperados durante el año 2006 en el Hospital "Eva Perón" de la Provincia de Buenos Aires. La caracterización fenotípica y genotípica de la resistencia permitió reconocer la presencia de metalo-beta-lactamasas (MBL) en el 14% de esos aislamientos. En todos ellos se identificó la presencia de la enzima IMP-13; sin embargo, algunos aislamientos resultaron sensibles a carbapenemes de acuerdo con los puntos de corte establecidos por el CLSI e incluso con las sugerencias de la Subcomisión de Antimicrobianos de SADEBAC, AAM. El ensayo de detección fenotípica de MBL de sinergia con doble disco resultó útil en este estudio. Sólo aquellos aislamientos productores de IMP-13 que a su vez presentaron alteraciones en las proteínas de membrana externa resultaron completamente resistentes a imipenem. Los aislamientos productores de MBL correspondieron a varios tipos clonales, lo cual sugiere no sólo la diseminación de una cepa resistente, sino también la diseminación horizontal de este mecanismo de resistencia entre clones diferentes.

From 129 P. aeruginosa isolated at a health care centre located in Buenos Aires (Hospital "Eva Perón"), 14% produced IMP-13. Although 18 isolates were metallo-beta-lactamases (MBL) producers, only those isolates that displayed altered outer membrane protein profiles correlated with the resistant category according to CLSI or even Subcomisión de Antimicrobianos, SADEBAC, AAM. Phenotypic screening of metallo-beta-lactamases proved to be appropriate for detecting MBL producing isolates. IMP-13 producing isolates corresponded to at least five different clonal types, which not only suggests the dissemination of the resistant strain but also of the resistant marker.

[Phenotypic and genotypic characterization of imipenem-resistant Pseudomonas aeruginosa isolated in a Buenos Aires hospital]. / Caracterización fenotípica y genotípica de la resistencia a imipenem en Pseudomonas aeruginosa aisladas en un hospital de Buenos Aires.

From 129 P. aeruginosa isolated at a health care centre located in Buenos Aires (Hospital "Eva Perón"), 14% produced IMP-13. Although 18 isolates were metallo-beta-lactamases (MBL) producers, only those isolates that displayed altered outer membrane protein profiles correlated with the resistant category according to CLSI or even Subcomisión de Antimicrobianos, SADEBAC, AAM. Phenotypic screening of metallo-beta-lactamases proved to be appropriate for detecting MBL producing isolates. IMP-13 producing isolates corresponded to at least five different clonal types, which not only suggests the dissemination of the resistant strain but also of the resistant marker.

[Comparison of different methods in order to identify Proteus spp]. / Comparación de diferentes métodos para identificar las especies del género Proteus.

Comparison of different methods in order to identify Proteus spp. The objectives were: (a) to identify Proteus strains to species level, following Farmer's and O'Hara's conventional biochemical reactions; b) to evaluate the sensitivity and specificity of both the API 20E method and a schema of reduced reactions (TSI and MIO agar: motility, indole and ornithine) comparing them with conventional methodology, and c) to evaluate the utility of SDS-PAGE (total proteins) in order to identify Proteus strains to species level. Two hundred and five Proteus spp. clinical isolates, were collected between January 1998 and September 2004, from inpatients and outpatients at Hospital de Clinicas. Strains were identified by means of conventional methodology, the API 20E method, and a schema of reduced reactions. SDS-PAGE (total proteins) was used in 48 out of the 205 strains. The API 20E method identified 79 out of 87 (90.8%) strains of P. mirabilis, 103 out of 103 P. vulgaris complex, and 15 out of 15 P. penneri. Eight strains of P. mirabilis were identified as Proteus spp., the acid production from maltose being necessary to identify them to species level. The schema of reduced reactions identified 205 out of 205 (100%) strains, that is, this schema of reduced reactions identified all the strains to species level without any additional tests, in marked contrast to the API 20E method. The SDS-PAGE (total proteins) identified the three species of the genus, even if the strains of P. mirabilis showed different biochemical reactions.

[Genetic environment of CTX-M-2 in Klebsiella pneumoniae isolates from hospitalized patients in Uruguay]. / Entorno genético de CTX-M-2 en aislamientos de Klebsiella pneumoniae provenientes de pacientes hospitalizados en Uruguay.

We studied two CTX-M-2-producing Klebsiella pneumoniae clinical strains, K96005 and K13, isolated from hospitalized patients in Uruguay, during 1996 and 2003, respectively. The genomic surroundings of bla(CTX-M-2) were characterized by PCR-mapping and DNA sequencing. Our results show that blaCTX-M-2 is included in a complex class-1 integron (InK13), associated with an orf513 in both isolates. The genetic array of the integron, aac(6')-lb, bla(OxA,2), orfD (gene cassette region), associated with an orf513-bla(CTX-M-2), seems to be widely disseminated over the Rio de la Plata region.

Comparación de diferentes métodos para identificar las especies del género Proteus / Comparison of different methods in order to identify Proteus spp

Los objetivos de este trabajo fueron: a) identificar a nivel de especie aislamientos de Proteus siguiendo la combinación de los esquemas de Farmer y O'Hara; b) determinar la utilidad del sistema comercial API 20E y de un esquema reducido de pruebas (agar TSI y agar MIO: movilidad, indol y ornitina), comparar estos procedimientos con la metodología convencional y evaluar su sensibilidad y especificidad, y c) evaluar la utilidad del perfil proteico en la identificación de las distintas especies. Se estudiaron 205 aislamientos de Proteus spp. aislados en el período comprendido entre enero de 1998 y setiembre de 2004, recuperados de distintos materiales clínicos correspondientes a pacientes hospitalizados y ambulatorios atendidos en el Hospital de Clínicas. Los organismos fueron identificados mediante la metodología convencional, por el sistema API 20E y con un esquema reducido de pruebas; 48 de ellos fueron sometidos a un SDS-PAGE. API 20E identificó 79 de 87 aislamientos de P. mirabilis (90,8%), 103/103 del complejo P. vulgaris y 15/15 de P. penneri. Ocho aislamientos identificados como Proteus spp. resultaron ser P. mirabilis, al incluir una prueba adicional (maltosa). En la identificación, el esquema reducido coincidió en un 100% con la metodología convencional. A diferencia del sistema API 20E, el esquema reducido alcanza la correcta identificación de todas las especies en laboratorios de baja complejidad, sin la necesidad de pruebas adicionales. El perfil proteico permitió la correcta diferenciación de las tres especies, independientemente de las diferentes atipias de P. mirabilis.

The objectives were: a) to identify Proteus strains to species level, following Farmer's and O'Hara's conventional biochemical reactions; b) to evaluate the sensitivity and specificity of both the API 20E method and a schema of reduced reactions (TSI and MIO agar: motility, indole and ornithine) comparing them with conventional methodology, and c) to evaluate the utility of SDS-PAGE (total proteins) in order to identify Proteus strains to species level. Two hundred and five Proteus spp. clinical isolates, were collected between January 1998 and September 2004, from inpatients and outpatients at Hospital de Clínicas. Strains were identified by means of conventional methodology, the API 20E method, and a schema of reduced reactions. SDS-PAGE (total proteins) was used in 48 out of the 205 strains. The API 20E method identified 79 out of 87 (90.8%) strains of P. mirabilis, 103 out of 103 P. vulgaris complex, and 15 out of 15 P. penneri. Eight strains of P. mirabilis were identified as Proteus spp., the acid production from maltose being necessary to identify them to species level. The schema of reduced reactions identified 205 out of 205 (100%) strains, that is, this schema of reduced reactions identified all the strains to species level without any additional tests, in marked contrast to the API 20E method. The SDS-PAGE (total proteins) identified the three species of the genus, even if the strains of P. mirabilis showed different biochemical reactions.

[Prevalence of metallo-beta-lactamase in carbapenem resistant Pseudomonas aeruginosa at a university hospital of Buenos Aires City]. / Prevalencia de metalo-beta-lactamasas en pseudomonas aeruginosa resistentes a carbapenemes en un hospital universitario de Buenos Aires.

The present study was conducted to estimate the prevalence of metallo-beta-lactamases in 91 consecutive carbapenem resistant Pseudomonas aeruginosa isolates, recovered from inpatients at Hospital de Clínicas in Buenos Aires. Both, phenotypic and genotypic methods detected the presence of carbapenemases in 10 (11%) isolates, corresponding to VIM-11 in 7/10 and VIM-2 in the others. Codifying genes were all included in class 1 integrons, upstream genes coding for aminoglycoside modifying enzymes. One hundred percent sensitivity and specificity was achieved by the metallo-beta-lactamases phenotypic screening method using EDTA (1 micromol) disks in the Pseudomonas aeruginosa isolates included in this study. Sensitivity to aztreonam in carbapenem resistant isolates was suspicious of the presence of these enzymes.

Predictors of the placebo response in functional dyspepsia.

BACKGROUND: Trials in functional dyspepsia report placebo response rates of 30% to 40%. AIM: We aimed to identify predictors of the placebo response. METHODS: Patients from primary, secondary and tertiary practices with functional dyspepsia defined by Rome II criteria were enrolled into one of four clinical trials; 220 patients were randomized to receive placebo. Scintigraphic assessment of gastric emptying at baseline was repeated at the end of the treatment in those with delayed emptying. After a 2 week run-in period, patients were followed for 8 weeks on placebo. Response was assessed on a weekly basis and a responder was defined as satisfactory relief of meal-related symptoms on at least 50% of weeks. RESULTS: The mean age was 44 years (range 18-82) and 74% were female; 76 (35%) were placebo responders. The predominant symptom was an unstable measure over the trial. Independent predictors of a lower placebo response were lower body mass index and a more consistent predominant symptom pattern (both P < 0.05). No association was seen with age, gender, centre type, baseline symptom score, baseline or change in gastric emptying, or baseline quality of life. CONCLUSION: In functional dyspepsia, a consistent predominant symptom pattern and lower body mass index may be associated with a lower placebo response rate.

Prevalencia de metalo-β-lactamasas en Pseudomonas aeruginosa resistentes a carbapenemes en un Hospital Universitario de Buenos Aires / Prevalence of metallo-β-lactamase in carbapenem resistant Pseudomonas aeruginosa at an University Hospital of Buenos Aires City

Se estudiaron 91 aislamientos de Pseudomonas aeruginosa resistentes a carbapenemes con el objetivo de conocer la prevalencia de metalo-β-lactamasas y evaluar la habilidad del ensayo de inhibición empleando discos de EDTA (1 µmol) en su detección. Se determinó la presencia de carbapenemasas en 10 (11%) de los aislamientos recuperados. La sensibilidad a aztreonam en los aislamientos resistentes a ambos carbapenemes resultó un buen predictor de la presencia de estas enzimas. Dichas carbapenemasas correspondieron a la enzima VIM-2 en tres de ellos y a VIM-11 en otros siete. En todos los casos los genes codificantes de estas enzimas se encontraron localizados en integrones de clase 1 seguidos corriente abajo de genes codificantes de enzimas acetilantes de antibióticos aminoglucosídicos. El ensayo de detección fenotípica de metalo-β-lactamasas empleando discos de EDTA mostró un 100% de especificidad y sensibilidad en la detección de estas enzimas en la población de Pseudomonas aeruginosa analizadas.

The present study was conducted to estimate the prevalence of metallo-β-lactamases in 91 consecutive carbapenem resistant Pseudomonas aeruginosa isolates, recovered from inpatients at Hospital de Clínicas in Buenos Aires. Both, phenotypic and genotypic methods detected the presence of carbapenemases in 10 (11%) isolates, corresponding to VIM-11 in 7/10 and VIM-2 in the others. Codifying genes were all included in class 1 integrons, upstream genes coding for aminoglycoside modifying enzymes. One hundred percent sensitivity and specificity was achieved by the metallo-β-lactamases phenotypic screening method using EDTA (1 µmol) disks in the Pseudomonas aeruginosa isolates included in this study. Sensitivity to aztreonam in carbapenem resistant isolates was suspicious of the presence of these enzymes.

Caracterización fenotípica y genotípica de la resistencia enzimática a las cefalosporinas de tercera generación en Enterobacter spp. / Phenotypic and genotypic characterization of resistance to third-generation cephalosporins in Enterobacter spp.

Enterobacter spp. es un patógeno intrahospitalario que presenta múltiples mecanismos de resistencia a los antibióticos b-lactámicos. Se caracterizaron fenotípica y genotípicamente las diferentes b-lactamasas presentes en 27 aislamientos consecutivos e ininterrumpidos de Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes). También se evaluó la habilidad de diferentes métodos fenotípicos para detectar b-lactamasas de espectro extendido (BLEE) en estos microorganismos. En 15/27 aislamientos (63%) se observó resistencia a las cefalosporinas de tercera generación. En 12 de los aislamientos resistentes se detectó un alto nivel de producción de cefalosporinasa cromosómica, siendo 6 de ellos también productores de PER-2. Dicha resistencia en los 3 aislamientos restantes se debió exclusivamente a la presencia de BLEE, PER-2 en 2 de ellos y CTX-M-2 en un caso. Sólo CTX-M-2 se detectó con todas las cefalosporinas probadas en los ensayos de sinergia, utilizando el método de difusión, mientras que cefepima mejoró la detección de PER-2 en 7/8 aislamientos productores de esta BLEE, 4/8 utilizando la prueba de doble disco y 7/8 comparando discos de cefepima con y sin el agregado de ácido clavulánico. El método de dilución empleado solo detectó 1/9 BLEE al comparar las cefalosporinas con y sin el agregado de inhibidor.

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to b-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum b-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63%) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC b-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor aproximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

Comprehension and awareness of symptoms in women with dyspepsia.

BACKGROUND: Overlapping symptoms of gastro-oesophageal reflux disease and dyspepsia are a problem for physicians and patients. AIM: This study explored comprehension of dyspepsia symptoms and associated medical terminology among women with symptoms of dyspepsia. METHODS: The US women aged > or = 18 years with dyspepsia (defined by Rome II criteria) were recruited in two phases, via direct mail, the Internet, clinical investigators and/or gastroenterologists. In phase I, subjects took part in an hour-long telephonic interview comprising open-ended questions relating to symptom frequency/duration, triggers/patterns and severity. During phase II, subjects took part in a 45-min telephonic interview, which explored their understanding of dyspepsia symptoms and their predominant or most bothersome symptom. RESULTS: Subjects with 'pure' dyspepsia (without overlapping symptoms of gastro-oesophageal reflux disease or irritable bowel syndrome) were sought, but of 777 subjects screened, most were excluded because of gastrointestinal comorbidities (irritable bowel syndrome, gastro-oesophageal reflux disease). Only 85 (11%) subjects had 'pure' dyspepsia of whom 11 withdrew. Of the 74 subjects interviewed, 70% were unfamiliar with the term 'dyspepsia'. Subjects reported several symptoms, including bloating (65%), gas (50%), nausea (41%) and discomfort (36%). Most subjects could distinguish between symptom bothersomeness and severity, and between pain and discomfort. Terms such as 'satisfactory relief', 'central upper abdominal discomfort', 'early satiety' and 'postmeal fullness' were often misunderstood. CONCLUSIONS: Subjects with 'pure' dyspepsia are rare, because of comorbidities. Dyspepsia-related terminology is often misunderstood by subjects.

[Consensus for antimicrobial susceptibility testing for Enterobacteriaceae. Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology]. / Consenso sobre las pruebas de sensibilidad a los antimicrobianos en Enterobacteriaceae.

Taking into account previous recommendations from the National Committee for Clinical Laboratory Standards (NCCLS), the Antimicrobial Committee, Sociedad Argentina de Bacteriología Clínica (SADEBAC), Asociación Argentina de Microbiología (AAM), and the experience from its members and some invited microbiologists, a consensus was obtained for antimicrobial susceptibility testing and interpretation in most frequent enterobacterial species isolated from clinical samples in our region. This document describes the natural antimicrobial resistance of some Enterobacteriaceae family members, including the resistance profiles due to their own chromosomal encoded beta-lactamases. A list of the antimicrobial agents that should be tested, their position on the agar plates, in order to detect the most frequent antimicrobial resistance mechanisms, and considerations on which antimicrobial agents should be reported regarding to the infection site and patient characteristics are included. Also, a description on appropriate phenotypic screening and confirmatory test for detection of prevalent extended spectrum beta-lactamases in our region are presented. Finally, a summary on frequent antimicrobial susceptibility profiles and their probably associated resistance mechanisms, and some infrequent antimicrobial resistance profiles that deserve confirmation are outlined.

Consenso sobre las pruebas de sensibilidad a los antimicrobianos en Enterobacteriaceae / Consensus for antimicrobial susceptibility testing for Enterobacteriaceae

En este documento se elaboraron una serie de recomendaciones para el ensayo, lectura, interpretación e informe de las pruebas de sensibilidad a los antimicrobianos para las enterobacterias aisladas con mayor frecuencia de especímenes clínicos. Se adoptaron como base las recomendaciones del National Committee for Clinical Laboratory Standards (NCCLS) de los EEUU, los de la subcomisión de Antimicrobianos, de la Sociedad Argentina de Bacteriología Clínica (SADEBAC), división de la Asociación Argentina de Microbiología (AAM) y de un grupo de expertos invitados. En él se indican las resistencias naturales de los diferentes miembros que integran la familia Enterobacteriaceae y se analiza la actividad de las diferentes beta-lactamasas cromosómicas, propias de cada especie, sobre las penicilinas, cefalosporinas y carbapenemes. Se recomiendan los antimicrobianos que se deberían ensayar, ubicados estratégicamente, para detectar los mecanismos de resistencia más frecuentes y cuales se deberían informar de acuerdo a la especie aislada, el sitio de infección y el origen de la cepa (intra o extrahospitalario). Se detallan los métodos de "screening" y de confirmación fenotipíca para detectar beta-lactamasas de espectro extendido (BLEE) que son más adecuados a nuestra realidad. Por último, se mencionan patrones infrecuentes de sensibilidad/resistencia que deberían verificarse y los perfiles de sensibilidad que pueden hallarse en las distintas enterobacterias en relación con los probables mecanismos de resistencia. Se debe resaltar que el contenido de este documento debe ser considerado como recomendaciones realizadas por expertos argentinos basadas en una revisión de la literatura y datos personales.

Taking into account previous recommendations from the National Committee for Clinical Laboratory Standards (NCCLS), the Antimicrobial Committee, Sociedad Argentina de Bacteriología Clínica (SADEBAC), Asociación Argentina de Microbiología (AAM), and the experience from its members and some invited microbiologists, a consensus was obtained for antimicrobial susceptibility testing and interpretation in most frequent enterobacterial species isolated from clinical samples in our region. This document describes the natural antimicrobial resistance of some Enterobacteriaceae family members, including the resistance profiles due to their own chromosomal encoded beta-lactamases. A list of the antimicrobial agents that should be tested, their position on the agar plates, in order to detect the most frequent antimicrobial resistance mechanisms, and considerations on which antimicrobial agents should be reported regarding to the infection site and patient characteristics are included. Also, a description on appropriate phenotypic screening and confirmatory test for detection of prevalent extended spectrum beta-lactamases in our region are presented. Finally, a summary on frequent antimicrobial susceptibility profiles and their probably associated resistance mechanisms, and some infrequent antimicrobial resistance profiles that deserve confirmation are outlined.

[Phenotypic and genotypic characterization of resistance to third-generation cephalosporins in Enterobacter spp]. / Caracterización fenotípica y genotípica de la resistencia enzimática a las cefalosporinas de tercera generación en Enterobacter spp.

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to beta-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63%) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC beta-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor approximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

Extended-spectrum beta-lactamases in enterobacteriaceae in Buenos Aires, Argentina, public hospitals.

Resistance to extended-spectrum cephalosporins is often associated with plasmid encoded extended spectrum beta-lactamases (ESBL). In order to evaluate the prevalence and diversity of ESBLs in enterobacteria in our city, a 1-month-period survey was carried out from April to May 2000. Extended-spectrum-cephalosporin-resistant strains, isolated from inpatient clinical specimens other than stools, were collected among 17 participating hospitals. From a total of 427 enterobacterial strains that were collected during this period, 39 were extended-spectrum cephalosporin resistant. The National Committee for Clinical Laboratory Standards' Screening and Confirmatory Tests for ESBL production were performed using cefotaxime and ceftazidime; cefepime and cefepime-clavulanic acid-containing disks were included. beta-Lactamases were characterized by isoelectric focusing and PCR amplification using specific primers. Three different ESBLs were detected: SHV-related (4 isolates), PER-2-type (9 isolates), and CTX-M-2-related (26 isolates). Sequencing of the corresponding genes confirmed CTX-M-2 in 19 of 21 and CTX-M-31 (an allelic variant) in the remaining 2 of 21. CTX-M-2 (or its variant) was detected in all Escherichia coli, Enterobacter aerogenes, Serratia marcescens, Proteus mirabilis, and Providencia stuartii strains, while PER-2 was detected in Enterobacter cloacae, E. aerogenes, and Klebsiella pneumoniae; SHV-related ESBL were found only in K. pneumoniae. These results clearly show that CTX-M-2 is the most prevalent ESBL produced by enterobacterial species isolated from public hospitals in Buenos Aires.

[Consensus on antimicrobial sensitivity tests in gram-positive cocci. Subcommittee on Antimicrobials, SADEBAC (Argentinian Society of Clinical Bacteriology), Argentinian Association of Microbiology]. / Consenso sobre las pruebas de sensibilidad a los antimicrobianos en cocos gram-positivos.

Antimicrobial susceptibility testing is mainly performed in Argentina by disk diffusion method, following National Committee for Clinical Laboratory Standards (NCCLS) recommendations. We worked out new recommendations for the reporting and interpretation of this test when dealing with gram-positive cocci, in accordance to local trends and epidemiology. General considerations for performing the diffusion assay, quality control, and an update on susceptibility testing for gram-positive cocci are reported in this first document. The present update should be considered as a group of recommendations summarized by Argentinean experts and as the result of a consensus meeting coordinated by the Subcomisión de Antimicrobianos of the Sociedad Argentina de Bacteriología Clínica (Asociación Argentina de Microbiología). Experts in antimicrobial agents were convened in order to prepare this final document. These recommendations take into account local needs, affordability and availability to be used in current practice, tending to contribute to the correct antimicrobial treatment election, according to the particular microorganism and the infection sites.

Consenso sobre las pruebas de sensibilidad a los antimicrobianos en cocos gram-positivos / Consensus for antimicrobial susceptibility testing for gram-positive cocci

El antibiograma por difusión en agar con discos se encuentra ampliamente difundido en nuestro medio y se basa primariamente en las recomendaciones del National Committee for Clinical Laboratory Standards (NCCLS). En este documento se elaboraron una serie de recomendaciones para el ensayo, lectura, interpretación e informe de las pruebas de sensibilidad a los antimicrobianos en cocos gram-positivos, adaptadas a la realidad argentina. En esta primera etapa se redactaron las consideraciones generales para la realización de la prueba por difusión, los controles de calidad internos para todos los microorganismos y una actualización sobre las pruebas de sensibilidad en cocos gram-positivos. Se debe resaltar que el contenido de este documento debe ser considerado como recomendaciones realizadas por expertos argentinos y que son el resultado de reuniones de consenso organizadas por la Subcomisión de Antimicrobianos de la Sociedad Argentina de Bacteriología Clínica, división de la Asociación Argentina de Microbiología. Se formó un equipo de trabajo integrado por expertos en antimicrobianos y a partir de una propuesta inicial, basada en una revisión de la literatura se fueron elaborando diversos documentos de trabajo que fueron mejorados después de ser debatidos por los miembros del grupo de trabajo hasta llegar al documento final. El criterio general fue elaborar recomendaciones acordes a las necesidades de nuestro país que puedan utilizarse en la práctica diaria con el objeto de colaborar en la adecuada elección del tratamiento antibiótico según la especie bacteriana aislada y la localización de la infección.

Key role of Shc signaling in the transforming pathway triggered by Ret/ptc2 oncoprotein.

The RET/PTC oncogenes, generated by chromosomal rearrangements in papillary thyroid carcinomas, are constitutively activated versions of protoRET, a gene encoding two protein isoforms of a transmembrane tyrosine kinase receptor. By using Ret/ptc2 short isoform (iso9), we have previously demonstrated that Tyr586 (Tyr1062 of protoRet) is the docking site for both the PTB and the SH2 domains of Shc. To determine the relevance of this interaction for the transforming activity of Ret/ptc oncogenes, we have generated and characterized novel Ret/ptc mutants unable to activate Shc: Ret/ptc2 long isoform (iso51)-Y586F and both isoforms of Ret/ptc2-N583A. These mutants neither activate Shc nor transform NIH3T3 cells. Since Tyr1062 shows features of a multifunctional docking site, we have used a Shc mutant (Shc Y317F) to directly assess Shc role. We have demonstrated that in our cell system Shc Y317F behaves like a dominant interfering mutant on the activation of the Grb2-Sos pathway by endogenous Shc triggered by Ret/ptc2. A strong reduction of the transforming activity of Ret/ptc2 in presence of this mutant was also demonstrated. Our data suggest that Shc activation play a key role in the transforming pathways triggered by Ret/ptc oncoproteins. Moreover, we have shown that coexpression of the Shc-Y317F mutant with Ret/ptc2 specifically causes apoptosis, and that the surviving cells lose the long-term expression of one of the two genes.