Acquisition of plasmids conferring carbapenem and aminoglycoside resistance and loss of surface-exposed macromolecule structures as strategies for the adaptation of Acinetobacter baumannii CC104O/CC15P strains to the clinical setting.

Acinetobacter baumannii (Aba) is an emerging opportunistic pathogen associated to nosocomial infections. The rapid increase in multidrug resistance (MDR) among Aba strains underscores the urgency of understanding how this pathogen evolves in the clinical environment. We conducted here a whole-genome sequence comparative analysis of three phylogenetically and epidemiologically related MDR Aba strains from Argentinean hospitals, assigned to the CC104O/CC15P clonal complex. While the Ab244 strain was carbapenem-susceptible, Ab242 and Ab825, isolated after the introduction of carbapenem therapy, displayed resistance to these last resource ß-lactams. We found a high chromosomal synteny among the three strains, but significant differences at their accessory genomes. Most importantly, carbapenem resistance in Ab242 and Ab825 was attributed to the acquisition of a Rep_3 family plasmid carrying a blaOXA-58 gene. Other differences involved a genomic island carrying resistance to toxic compounds and a Tn10 element exclusive to Ab244 and Ab825, respectively. Also remarkably, 44 insertion sequences (ISs) were uncovered in Ab825, in contrast with the 14 and 11 detected in Ab242 and Ab244, respectively. Moreover, Ab825 showed a higher killing capacity as compared to the other two strains in the Galleria mellonella infection model. A search for virulence and persistence determinants indicated the loss or IS-mediated interruption of genes encoding many surface-exposed macromolecules in Ab825, suggesting that these events are responsible for its higher relative virulence. The comparative genomic analyses of the CC104O/CC15P strains conducted here revealed the contribution of acquired mobile genetic elements such as ISs and plasmids to the adaptation of A. baumannii to the clinical setting.

Agrobacterium-mediated transformation of tomato with the ICE1 transcription factor gene.

ICE1 genes play a very important role in plants in cold conditions. To improve the cold resistance of tomato, the ICE1 gene of Arabidopsis thaliana was used to construct the plant expression vector p3301-ICE1, and was overexpressed in tomato through Agrobacterium-mediated transformation. Five strains of resistant plants were obtained. PCR and half-quantitative results showed that the ICE1 gene was transferred to tomato; three strains tested positive. After low-temperature stress treatment, praline content and peroxide and catalase activities in the transgenic tomato plants were higher compared with non-transgenic controls, while malondialdehyde content was clearly lower.

Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes.

Micro-Tom is the smallest known variety of tomatoes. An orthogonal experimental design L(16) (4(5)) was used to optimize Agrobacterium-mediated transformation of cotyledon explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were investigated to determine their effect on transformation frequency: the concentration of bacterial suspension, time of dip in bacterial suspension, co-cultivation time, and concentration of carbenicillin. We also examined the effect of these parameters on contamination rate, necrosis rate, mortality, cut-surface browning rate, and undamaged explant rate. Both the bacterial and carbenicillin concentrations had a significant influence on the rate of infected explants. The time of co-cultivation also had a significant influence on the transformation parameters. The optimal transformation protocol consisted of an Agrobacterium suspension of 0.5 × 10(8) cells/mL (OD(600) = 0.5) and an infection time of 5 min, one day of co-cultivation and 500 mg/L carbenicillin. Under these conditions, the transformation efficiency of the shoots reached 5.1%; the mean transformation frequency was 3.9% (N = 838).

Improved production of transgenic Dioscorea zingiberensis (Dioscoreaceae) by Agrobacterium tumefaciens-mediated transformation.

The establishment of high-efficiency Agrobacterium-mediated transformation techniques could improve the production of Dioscorea zingiberensis, a medicinal species with a high diosgenin content. We co-cultivated embryogenic calli induced from mature seeds with A. tumefaciens strain EHA105. A binary vector, pCAMBIA1381, which contains the gfp and hpt genes under the control of the ubiquitin promoter and the CaMV 35S promoter, respectively, was used for transformation. Pre-culture, basic medium, acetosyringone, and bacterial density were evaluated to establish the most efficient protocol. The optimal conditions consisted of MS medium without CaCl(2) for pre- and co-cultivation, three days for pre-culture, addition of 200 µM AS, and an OD(600) of 0.5. The transgenic plants grown under selection were confirmed by PCR analysis and Southern blot analysis. This protocol produced transgenic D. zingiberensis plants in seven months, with a transformation efficiency of 6%.

[Bacterial meningitis and Pseudomonas aeruginosa: apropos of a case]. / Meningitis bacteriana y Pseudomonas aeruginosa: a propósito de un caso.

Meningitis caused by gram-negative bacilli increased since the 1970, with a higher incidence in small children. Within this group of infections, the meningitis caused by Pseudomonas sp is rare. The case of a 54-year-old patient with a clinical picture of meningitis is reported. Pseudomonas aeruginosa was isolated from the cerebrospinal fluid. The meningitis caused by Pseudomonas aeruginosa should be taken into consideration because of the severity of the clinical picture and the high mortality and increasing strain resistance.

New carbenicillin-hydrolyzing beta-lactamase (CARB-7) from Vibrio cholerae non-O1, non-O139 strains encoded by the VCR region of the V. cholerae genome.

In a previous study, an analysis of 77 ampicillin-nonsusceptible (resistant plus intermediate categories) strains of Vibrio cholerae non-O1, non-O139, isolated from aquatic environment and diarrheal stool, showed that all of them produced a beta-lactamase with a pI of 5.4. Hybridization or amplification by PCR with a probe for bla(TEM) or primers for bla(CARB) gene families was negative. In this work, an environmental ampicillin-resistant strain from this sample, ME11762, isolated from a waterway in the west region of Argentina, was studied. The nucleotide sequence of the structural gene of the beta-lactamase was determined by bidirectional sequencing of a Sau3AI fragment belonging to this isolate. The gene encodes a new 288-amino-acid protein, designated CARB-7, that shares 88.5% homology with the CARB-6 enzyme; an overall 83.2% homology with PSE-4, PSE-1, CARB-3, and the Proteus mirabilis N29 enzymes; and 79% homology with CARB-4 enzyme. The gene for this beta-lactamase could not be transferred to Escherichia coli by conjugation. The nucleotide sequence of the flanking regions of the bla(CARB-7) gene showed the occurrence of three 123-bp V. cholerae repeated sequences, all of which were found outside the predicted open reading frame. The upstream fragment of the bla(CARB-7) gene shared 93% identity with a locus situated inside V. cholerae's chromosome 2. These results strongly suggest the chromosomal location of the bla(CARB-7) gene, making this the first communication of a beta-lactamase gene located on the VCR island of the V. cholerae genome.

Empirical antimicrobial therapy in pediatric patients with neutropenia and fever. Risk factor for treatment failure

Background. The use of combinations of antibiotics has been the cornerstone of therapy for febrile patients with cancer and severe neutropenia. Each empirical regimen should be selected according to the epidemiology and susceptibility patterns in each center. We describe here the experience wtih empirical antimicrobiial treatments in pediatric patients with cancer, fever and severe neutropenia, and identify the risk factors associated with treatment failure. Methods. This is a prospective study including 145 patients with cancer, and 171 episodes of neutropenia and fever. Blood cultures were taken before initiating empirical treatment: a)carbenicillin (400 mg/kg/day) plus amikacin (21 mg/kg/day) (Cb/ak), and b) ceftazidime (100 mg/kg/day), plus amikacin at the same dosage (Cz/ak). Results. The overall response rate was 54.9 percent and 56.3 percent for Cb/ak and Cz/ak, respectively. Fifty-seven episodes (33.3 percent) were microbiologically documented, gram-positive isolated in 38 percent and gram-negative in 49 percent. Risk factors associated significantly with treatment failure were acute mywlocytic leukemia (AML) (RR 2.59, CI 95 percent 1.42-4.7, p=0.003); bacteriological identification (RR= 4.41, CI 95 percent 2.21 - 8.8, p<0.001), and the presence of two or more sites of infection (RR= 2.89, CI 95 percent 1.03 - 8.11, p=0.03). Conclusions. The rates of response are similar to the combinations used in the hospital (Cb/ak, Cz/ak). The risk factors associated with treatment failure were AML diagnosis, bacteriological identification, and the presence of two or more sites of infection

Serotypes of Vibrio cholerae non-O1 isolated from water supplies for human consumption in Campeche, México and their antibiotic susceptibility pattern.

The presence of Vibrio cholerae non-O1 in water supplies for human consumption in the city of Campeche and rural locality of Bécal was investigated. V. cholerae non-O1 was detected in 5.9% of the samples obtained in deep pools of Campeche. Studies conducted in Bécal and neighbourhood of Morelos in Campeche indicated that collected samples harbored V. cholerae non-O1 in 31.5% and 8.7% respectively. There was a particular pattern of distribution of V. cholerae non-O1 serotypes among different studied regions. Accordingly, V. cholerae non-O1 serotype O14 predominated in the deep pools of Campeche and together with V. cholerae non-O1, O155 were preferentially founds in samples taken from intradomiciliary faucets in the neighbourhood of Morelos. Samples from Bécal predominantly presented the serotype O112. 60% and 53.8% of all studied strains of V. cholerae non-O1 proved to be resistant to ampicillin and carbenicillin. 3.1%, 7.7% and 6.2% presented resistant to doxycycline, trimethoprim-sulfamethoxazole and erythromycin respectively. The study showed the necessity of performing a strong epidemiologic surveillance for emergence and distribution of V. cholerae non-O1.

Amikacina en dosis única diaria en niños neutropénicos con fiebre / Amikacin once-daily in febrile neutropenic children

Se realizó un ensayo clínico aleatorizado en niños con cáncer, fiebre y neutropenia para evaluar la eficacia de la amikacina en dosis única diaria contra tres dosis al día asociada a carbenicilina en ambos grupos. Se incluyeron 50 pacientes, 25 pacientes en el grupo A que recibieron la dosis diaria única de amikacina, y 25 pacientes en el grupo B que recibieron la amikacina fraccionada en tres dosis cada 8 horas. No hubo diferencias intergrupos: la fiebre remitió en una mediana de 6 días (2-8días) vs 7 días (3-12 días) en los grupos A y B, respectivamente (p= 0.37); la mejoría clínica se observo en una mediana de 6 días (3-10 días) vs 7 días (2-14 días) (p=0.68). Un paciente en el grupo A y dos en el B fallecieron. Los picos máximos de amikacina al 7o. día de tratamiento fueron de 10-60 y de 7-25 µg/mL en los grupos A y B respectivamente, y los niveles séricos de creatinina de 0.3-0.7 mg/dL para el grupo A y de 0.2 - 0.8 mg-dL para el grupo B; ningún paciente presentó elevación de la creatinina mayor al 40 por ciento del basal. Tres pacientes del grupo A tuvieron niveles de amikacina mayores a 40 µg/mL sin que existiera elevación de la creatinina. No encontramos datos que sugieran que la toxicidad fue mayor. La modalidad de administración de aminoglucósido en una dosis al día parece ser igual de efectiva que la convencional

Randomized trial comparing oral ciprofloxacin plus penicillin V with amikacin plus carbenicillin or ceftazidime for empirical treatment of febrile neutropenic cancer patients.

Aminoglycoside-containing combination therapy has been the standard empirical approach for febrile neutropenic cancer patients. With the advent of the broad-spectrum oral fluoroquinolones, it is now possible to evaluate an initial empirical alternative therapy. A prospective randomized study was conducted comparing oral ciprofloxacin plus penicillin V (group A) with amikacin plus carbenicillin or ceftazidime (group B). Main criteria for eligibility were febrile patients with solid tumor or nonlymphoblastic lymphoma, a Zubrod PS equal to 1 or 2, no diarrhea, mucositis, or long-term central venous catheter. A total of 108 consecutive neutropenic febrile episodes were randomized (5 exclusions); 55 episodes were assigned to group A and 48 to group B. Most febrile episodes were of unknown origin. There were 10 microbiologically documented episodes with two cases of bacteremia. Both regimens were well tolerated. Oral regimen was substantially cheaper than parenteral regimen. Treatment success without regimen modification was 94.5% for group A and 93.8% for group B (p = .86; CI -0.08-0.10). Oral therapy with ciprofloxacin and penicillin V is a safe alternative to standard parenteral therapy in this low-risk group of neutropenic patients, with unquestionable cost containment.

Serovariedades de Salmonella em águas superficiais em San Luís, Argentina / Salmonella serotypes from surface waters in San Luis, Argentina

Resumo: Foi investigada a presença de Smonella em amostras de água colhidas pela técnica de Moore, em rios e açudes, durante um ano. Realizau-se um pré-enriquecimento em água peptomada tamponada e um enriquecimento em caldo tetrationato-verde brilhante com caldo Rappaport. Isolou-se em 0gar verde brilhantre com desoxicolato de sódio a 0,25 (por cento) e em ágar sulfito de bismuto. As colônias suspeitas foram estrudadas mediante provas bioqu1micas clássicas e sorológicas. Recuperou-se Salmonella em 6,66 (por cento) das amostras. Isolou-se S. panama, S. newport, S. sandiego e Salmonella sub-espécie IV, 40: Z4, Z23: esta última serovariedade constitui o primeiro isolamento na Argentina. Determinou-se o antibiograma pelo método de Kirby-Bauer. As 15 cepas isoladas foram sensíveis a trimetroprima-sulfametoxazol, gentamicina, tetraciclina, cloranfenicol, kamamicina e ácido nalidíxico, quanto à carbenicilina, 6 cepas foram resistentes, 7 mostraram sensibilidade intermediária e só 2 foram sensíveis. -Foram determinados perfis plasmídicos pela técnica de Birnboin e dOLY. Os pesos moleculares (PM) dos plasmídeos em megadaltons (MDa) foram determinados por comparaçäo com plasmídeos de PM conhecidos. Foram detectados plasmídeos de PM 40 MDa em S. newpot e de PM 17 MDa em S. sandiego. Seria de interesse a determinaçäo sistemática de marcadores epidemiológicos em surtos e casos esporádicos de salmonelose com o fim de estabelecer sua distribuiçäo geográfica na zona estudada da província de San Luis, assim como também a necessidade de uma vigilância microbiológica das águas superficiais e a implementaçäo de tratamentos de depuraçäo de águas cloacais adequadois antes de serem despejadas nos cursos de água (au)

Peritonitis por Serratia marcescens en un paciente sometido a diálisis peritoneal contínua ambulatoria / Serratia marcescens peritonitis in a patient under continous ambulatory peritoneal dialysis

El objetivo del presente trabajo es la exposición de lo que se considera el primer caso de peritonitis en un paciente con diálisis peritoneal continua ambulatoria (DPCA) -por insuficiencia renal crónica terminal-causada por Serratia marcescens y tratado exitosamente con carbenicilina. Se trata de un bacilo gramnegativo, móvil, aerobio, miembro de la división Klebsiella-Enterobacter-Serratia, entre la familia enterobacteriaceae. La Serratia contrasta con otros bacilos enterobacteriaceos en que no es huésped habitual del tracto digestivo y se adquiere por contaminación iatrógena como manipulación genitourinaria, diálisis peritoneal, hemodialisis y punción lumbar. Su tratamiento antibiótico es difícil pues presenta gran resistencia medicamentosa por plásmidos

The treatment of acute myeloid leukemia in Brazil: progress and obstacles.

BACKGROUND: Substantial progress has been made in the treatment of acute myeloid leukemia in the last two decades. We wanted to evaluate the outcome of intensive chemotherapy and the influence of recent therapy changes in underprivileged patients treated in a large urban public university hospital. METHODS: The records of all patients treated for acute myeloid leukemia from 1980 to 1993 were analyzed. RESULTS: 109 patients were identified; 41 did not receive any treatment for the leukemia because of infectious and/or hemorrhagic complications of advanced disease. Median survival in this group was 4 days. The other 68 patients received one of two induction protocols: TAD from 1980 to 1985 (n = 23) and ara-C plus daunorubicin from 1985 to 1992 (n = 45). The complete remission rate was 56%, disease-free survival 24% and overall survival 15% at 13 years. Overall survival was better for patients treated with ara-C plus daunorubicin than with TAD (19% versus 8%, p = 0.01). This is attributed to a reduction in infection mortality after ceftazidime and amikacin replaced cephalotin, carbenicillin and amikacin as the antibiotic regimen. CONCLUSIONS: The most effective intervention in our population would probably be an improvement in the primary health care system, so that earlier diagnosis could allow the treatment of a larger fraction of patients.

Otite externa maligna: diagnóstico e tratamento precoces: relato de um caso clínico / Malignant otitis externa: diagnosis and treatment

A otite externa maligna é uma entidade de prognóstico reservado, quando seu diagnóstico é tardio e a conduta terapêutica apropriada näo é entäo instituida. O autor relata um caso documentado de paciente diabético, obtendo-se em sua propêudica o diagnóstico final. O tratamento foi realizado com o paciente internado durante 32 dias no HSE. O prognóstico da enfermidade em questäo indica êxito letal de 53 por cento em várias casuísticas. Felizmente neste caso o resultado foi favorável com a terapêutica instituída, que incluiu equipe multidisciplinar, levando à cura

Salmonella em raçäo para aves no Brasil / Salmonella in poultry feeds in Brazil

This experiment was undertaken to determine the possible presence of Salmonella in poultry diets.A total of two hundred samples of ration fr4om 4 commercial poultry feed industries were examined.The results revealed the presence of salmonellae in 10 cents of the samples studied and 14 serotypes were identified.The produce for Salmonella isolation included the pre-enrichment step and the strains were submited to antimicrobial tests.The 29 strains were resistant to the followings antimicrobial agents(porcents

Yersinia spp: Estudo das características biológicas das amostras isoladas de queijo tipo Minas Frescal / Yersinia spp: a study of the biological characteristics of strains isolated fromm minas frescal cheese (white cheese)

resumo: foram analisadas 50 amostras de dez diferentes marcas comerciais de queijo tipo "Minas Frescal", comercializados em supermecados do Rio de Janeiro.Foram pesquisados os microrganismos indicadores de contaminaçäo fecal pelo método do número mais provável.Os coliformes totais variaram de 2, 7*10elevado a terceira/g a 2, 4*10elevado a sexta/g e os coliformes fecais de 2*10/g a 2, 1*10elevado a sexta/g.Das amostras examinadas, 80 por cento mostraram precárias condiçöes higiênico-sanitárias, de acordo com os limites estabelecidos pelos padröes do Ministério da Saúde(DNVS).paralelamente, foram pesquisaas Yersinia spp por três diferentes metodologias.o método do pré-enriquecimento a 26ºC/48 horas e posterior tratamento alcalino sequido de semeadura em agar desoxicolato citrato permitiu a identificaçäo de 11 amostras de Y.frederiksenii-0:16a, 16b-Xo.Pelo método do plaqueamento direto, apenas uma amostra de Y.intermedia-NAG-Xo no meio agar por 7 a 21 dias näo permitiu o isolamento de nenhuma amostra.Essas amostras de yersinia spp foram submetidas aos testes de produçäo de enterotoxina termoestável, autoaglutinaçäo, cálcio-dependência, capacidade de ligaçäo ao cristal violeta, atividade de piraminamidase, atividade enzimática extracelular, com objetivo de avaliar o seu comportamento biológico.Todas se comportam negativamente, com exceçäo de uma amostra de Y.frederiksenii que foi protease positiva.Finalmente, o perfil de sensibilidade das Yersinia spp isoladas, mosxtrou uma multiresistência em relaçäo à vários antimicrobianos.Apresentaram uma melhor sensibilidad ao ácido nalidíxico, amicacina, cloranfenicol, gentamicina, norfloxacina, polimixina B e Tobramicina.Todas as amostras de Yersinia spp isoladas no presente estudo tiveram a capacidade de produzir B-lactamase, através do método de Nitrocefin.

Iatrogenia en neurología / Iatrogenesis in neurology

LAs iatrogenias más frecuentes en Neurología son: reacciones adversas y enfermedades inducidas por medicamentos, complicaciones de técnicas diagnósticas, complicaciones de procederes quirúrgicos y complicaciones de métodos terapéuticos (AU)