In vitro antagonism of rabeprazole and metronidazole upon clinical isolates of Helicobacter pylori.

BACKGROUND: Rabeprazole inhibits Helicobacter pylori in vitro. This property could influence the antibacterial activity of metronidazole or amoxicillin. Our goal was to evaluate in vitro the effect of rabeprazole upon the antibacterial activity of metronidazole and amoxicillin. METHODS: The minimal inhibitory concentration (MIC) was determined by agar dilution upon 30 clinical isolates, and the antibacterial effect of the combined drugs was estimated by calculating the fractional inhibitory concentration index (FICI). RESULTS: Nine strains (30%) were resistant to metronidazole (MIC 8-256 mg/l) while only 1 strain was resistant to amoxicillin (MIC 1.0 mg/l). Rabeprazole also inhibited H. pylori (MIC 0.125-1.0 mg/l). The association of metronidazole and rabeprazole showed synergism in 30% of the strains (FICI < or =0.25), but also antagonism in 23.3% of the clinical isolates (FICI > or =4.0). Mostly, amoxicillin-rabeprazole showed an effect of synergism (53.3%). CONCLUSION: The results presented suggest the need to keep in mind that failure in eradication therapies based on metronidazole and rabeprazole could be due to an antagonism of the drugs.

Carbapenem-hydrolysing beta-lactamase KPC-2 in Klebsiella pneumoniae isolated in Rio de Janeiro, Brazil.

OBJECTIVES: The aim of this study was to characterize the KPC-type carbapenem-hydrolysing beta-lactamase, extended-spectrum beta-lactamases (ESBLs) and class 1 integrons among nosocomial Klebsiella pneumoniae isolated in Rio de Janeiro, Brazil. METHODS: MICs were determined and isolates were screened for ESBLs, metallo-beta-lactamases (MBLs) and class A carbapenemase-producing phenotypes. The main beta-lactamases resistance genes (bla(TEM), bla(SHV), bla(CTX-M), bla(KPC), bla(IMP) and bla(VIM)) and class 1 integrons were detected by PCR followed by DNA sequencing. The genetic relatedness of isolates was determined by PFGE. RESULTS: All K. pneumoniae isolates were positive for ESBL and class A carbapenemase production and negative for MBL production. All isolates were resistant to all beta-lactam antibiotics, ciprofloxacin and gentamicin, being susceptible only to tigecycline and polymyxin B. The bla(KPC-2), bla(CTX-M-1), bla(CTX-M-2), bla(CTX-M-8) and bla(SHV-11) genes were detected. PFGE analysis revealed two clonal types among KPC-producing isolates, both identified in the same hospital. CONCLUSIONS: Our findings should alert medical authorities to implement stringent methods for the detection and spread control of emerging KPC-2 carbapenemases in the hospital setting in Brazil.

Bacteriocin-like inhibitory substances from Campylobacter spp.

Twenty-five Campylobacter isolates were screened for production of antimicrobial substances using a deferred antagonism assay. Sixteen isolates showed activity against either Staphylococcus aureus, Salmonella enterica serovar Enteritidis or Candida albicans. The inhibitory activity was sensitive to treatment with pronase E, trypsin and pepsin, suggesting that the antimicrobial compound(s) are proteinaceous. Activity spectra of isolates included S. aureus, Micrococcus luteus, Streptococcus sp., Bacillus subtilis, a drug-resistant clinical isolate of S. aureus and one isolate of C. albicans. Producing isolates showed cross-immunity and inhibitory activity was only observed on solid media. The findings of this study suggest that Campylobacter produces proteinaceous inhibitory substances.

Antagonistic properties of some microorganisms isolated from Brazilian tropical savannah plants against Staphylococcus coagulase-positive strain

Endophytic microorganisms are relatively unstudied as potential sources of novel natural products for medical and commercial exploitation. The aim of this work was to investigate some Brazilian tropical savannah trees Cassia leptophylla and Prunus spp. in order to isolate the endophytic microorganisms associated with these plants. The samples were disinfected to eliminate the epiphytic population. Colonies were diluted and displayed as drops in media and growing colonies were inactivated. Staphylococcus coagulase-positive strain was used as indicator microorganism and subjected to the antibioses test. Data showed that the microorganisms isolated from Cassia leptophylla had no inhibition against Staphylococcus. On the other hand, microorganisms isolated from Prunus spp. leaves showed antibacterial activity and inhibited Staphylococcus when cultivated in peptone agar as well as in yeast extract agar. Investigation proceeds in order to classify the isolated microorganisms presenting bioactive substance and exploit the potential of the compounds produced to inhibit the indicator bacteria. Other bioactive properties will be investigated.

Infección relacionada con el drenaje ventricular externo: tratamiento intravenoso versus intratecal, revisión de la literatura / External ventricular drainage infection: intravenous vs intrathecal treatment, Review

Ante la emergencia de infecciones relacionadas al drenaje ventricular externo por gérmenes multirresistentes y la necesidad de utilizar antibióticos cuya acción en el sistema nervioso central es controvertida, surge la necesidad de emplear terapéuticas para la administración de fármacos cuyos beneficios merecen ser evaluados. Se realizó una búsqueda de la literatura en PUBMED y los datos fueron extraídos de estudios publicados entre 1987-2005. En esta revisión de la literatura se analizaron los resultados obtenidos, comparando: el tratamiento endovenoso con el tratamiento intratecal en pacientes con infección asociada a la ventriculostomía, los antibióticos utilizados y su penetración al líquido cefalorraquídeo (LCR) con cada vía. Finalmente, se propuso qué pacientes podrían beneficiarse con cada terapéutica, a pesar de la falta de evidencias claras, sugiriendo la necesidad de realizar un trabajo randomizado prospectivo comparando sus beneficios. Palabras clave: infección, tratamiento, ventriculitis.

Due to the emergency of external ventricular drainage infections, cause by multiresistant bacterial strains, and the need to treat them with antibiotics, whose action in the central nervous system is controversial, arises the terapeutic necessity to administer antibiotics whose benefits should be evaluated. We search the Pubmed library. The data were extracted from published studies between 1987-2005. In this reivision the results were analyzed, comparing envovenous versus intrathecal treatment in patients with a ventriculostomy associated infection, the antibiotics that were used and their cerebrospinal fluid (CSF) levels according to each route. Finally, even though there is a lack of clear evidence, we proposed which patients could benefit with each treatment. We conclude that a randomized prospective study, comparing the benefits of each treatment is needed.

Infección relacionada con el drenaje ventricular externo: tratamiento intravenoso versus intratecal, revisión de la literatura / External ventricular drainage infection: intravenous vs intrathecal treatment, Review

Ante la emergencia de infecciones relacionadas al drenaje ventricular externo por gérmenes multirresistentes y la necesidad de utilizar antibióticos cuya acción en el sistema nervioso central es controvertida, surge la necesidad de emplear terapéuticas para la administración de fármacos cuyos beneficios merecen ser evaluados. Se realizó una búsqueda de la literatura en PUBMED y los datos fueron extraídos de estudios publicados entre 1987-2005. En esta revisión de la literatura se analizaron los resultados obtenidos, comparando: el tratamiento endovenoso con el tratamiento intratecal en pacientes con infección asociada a la ventriculostomía, los antibióticos utilizados y su penetración al líquido cefalorraquídeo (LCR) con cada vía. Finalmente, se propuso qué pacientes podrían beneficiarse con cada terapéutica, a pesar de la falta de evidencias claras, sugiriendo la necesidad de realizar un trabajo randomizado prospectivo comparando sus beneficios. Palabras clave: infección, tratamiento, ventriculitis. (AU)

Due to the emergency of external ventricular drainage infections, cause by multiresistant bacterial strains, and the need to treat them with antibiotics, whose action in the central nervous system is controversial, arises the terapeutic necessity to administer antibiotics whose benefits should be evaluated. We search the Pubmed library. The data were extracted from published studies between 1987-2005. In this reivision the results were analyzed, comparing envovenous versus intrathecal treatment in patients with a ventriculostomy associated infection, the antibiotics that were used and their cerebrospinal fluid (CSF) levels according to each route. Finally, even though there is a lack of clear evidence, we proposed which patients could benefit with each treatment. We conclude that a randomized prospective study, comparing the benefits of each treatment is needed. (AU)

Infección relacionada con el drenaje ventricular externo: tratamiento intravenoso versus intratecal, revisión de la literatura / External ventricular drainage infection: intravenous vs intrathecal treatment, Review

Ante la emergencia de infecciones relacionadas al drenaje ventricular externo por gérmenes multirresistentes y la necesidad de utilizar antibióticos cuya acción en el sistema nervioso central es controvertida, surge la necesidad de emplear terapéuticas para la administración de fármacos cuyos beneficios merecen ser evaluados. Se realizó una búsqueda de la literatura en PUBMED y los datos fueron extraídos de estudios publicados entre 1987-2005. En esta revisión de la literatura se analizaron los resultados obtenidos, comparando: el tratamiento endovenoso con el tratamiento intratecal en pacientes con infección asociada a la ventriculostomía, los antibióticos utilizados y su penetración al líquido cefalorraquídeo (LCR) con cada vía. Finalmente, se propuso qué pacientes podrían beneficiarse con cada terapéutica, a pesar de la falta de evidencias claras, sugiriendo la necesidad de realizar un trabajo randomizado prospectivo comparando sus beneficios. Palabras clave: infección, tratamiento, ventriculitis. (AU)

Due to the emergency of external ventricular drainage infections, cause by multiresistant bacterial strains, and the need to treat them with antibiotics, whose action in the central nervous system is controversial, arises the terapeutic necessity to administer antibiotics whose benefits should be evaluated. We search the Pubmed library. The data were extracted from published studies between 1987-2005. In this reivision the results were analyzed, comparing envovenous versus intrathecal treatment in patients with a ventriculostomy associated infection, the antibiotics that were used and their cerebrospinal fluid (CSF) levels according to each route. Finally, even though there is a lack of clear evidence, we proposed which patients could benefit with each treatment. We conclude that a randomized prospective study, comparing the benefits of each treatment is needed. (AU)

Investigation of genotoxic and antigenotoxic activities of Melampodium divaricatum in Salmonella typhimurium.

Melampodium divaricatum is a member of the Asteraceae and in Brazil is known as false-calendula, its flowers being used in anti-inflammatory preparations, substituting the true calendula or marigold (Calendula officinalis L.). The flower extract was investigated for mutagenic and antimutagenic effect in the Salmonella/microsome assay. The tested extract was not mutagenic in the strains TA100, TA98, TA97a and TA102 and decreased the mutagenicity of aflatoxin B1, benzo(a)pyrene and daunomycin. Chlorophyll and triterpenes were detected in the extract, and they might have contributed to the observed effect. Our data suggest that these medicinal plants possess cancer chemopreventive properties.

Aislamiento de plásmidos conjugativos en cepas de enterobacterias productoras de ß-lactamasas de espectro expandido / Isolation of conjugative plasmids in enterobacteriaceae strains producing beta-lactamases expanded sprectrum

Las ß-lactamasas de espectro expandido son enzimas capaces de hidrolizar el enlace amida de los oximino ß-lactámicos, (cefotaxime, ceflazidime y aztreonam). Las mismas son codificadas en plásmidos y por lo tanto pueden ser transferidas mediante conjugación a diversos géneros bacterianos. Diseminándose ampliamente en el ambiente hospitalario. En esta investigación se persigue, en cepas de enterobacterias aislar plásmidos que codifican para ß-lactamasas de espectro expandido y que sean capaces de transferirse mediante conjugación. Se estudio una población de 51 enterobacterias productoras de ß-lactamasas de espectro expandido aisladas de diferentes centros hospitalarios del área Metropolitana de Caracas. A las mismas se le determinó el perfil de resistencia a múltiples antibióticos mediante la metodología de Kirby-Bauer. Se detectaron las BLEE mediante dos ensayos fenotípicos basados en el efecto sinergístico con el ácido clavulánico y se tipificaron molecularmente por PCRIRFLP, seguidamente se transfirieron los plásmidos conjugativos en ensayos de conjugación en medio sólido y se aislaron los plásmidos de cepas donantes y transconjugautes, por el método de lisis alcalina. Los resultados fenotípicos indican una mayor proporción de BLEE con actividad ceflazidimasa y en menor grado actividad cefotaximasa. La tipificación molecular indicó que 60,8 por ciento de las cepas portan genes tipo SHV y 15,6 por ciento codifican ß-lactamasas de espectro expandido de la familia CTX-M. De 36 cepas conjugadas un 81 por ciento transfirió material plasmídico. El análisis de los aislamientos plasmídicos mostró la presencia en la totalidad de las transcojugantes de una banda de 25000 pb y en un 80 por ciento se evidenció una banda plasmídica mayor a 50000 pb. Se pudo constatar la cotransferencia de resistencia a otras familias de antibióticos.

Detection of a chloramphenicol efflux system in Escherichia coli isolated from poultry carcass.

An active chloramphenicol efflux system was demonstrated in a multiresistant E. coli isolated from poultry carcass. The effect of different concentrations of chloramphenicol on the original strain and on the plasmid-cured strain was determined in the presence and in the absence of CCCP, an uncoupler of the proton-motive force. Minimal inhibitory concentration (MIC) was lower in the presence of CCCP in the original strain. The plasmid-cured strain displayed lower resistance for chloramphenicol than the wild type, but the MIC was not affected by CCCP. The combined results indicate a plasmid encoded energy dependent resistance mechanism. 3H-chloramphenicol accumulation within the cells was measured by scintillation counting. The uptake or the efflux of 3H-chloramphenicol was influenced by CCCP in the original strain, but not in the plasmid-cured strain. More than one chloramphenicol resistance mechanism may exist in this strain. E. coli is an important commensal or pathogen that inhabits the gastrointestinal tracts of humans and animals, so a plasmid encoded active drug resistance mechanism can be a potential source of horizontal transfer of resistance.

Aged garlic extract attenuates gentamicin induced renal damage and oxidative stress in rats.

Gentamicin (GM) is an antibiotic whose clinical use is limited by its nephrotoxicity. Experimental evidences suggest a role of reactive oxygen species in GM-induced nephrotoxicity. Therefore, we investigated if aged garlic extract (AGE), an antioxidant, has a protective role in this experimental model. Four groups of male Wistar rats were studied: 1) Control (CT), injected subcutaneously (s.c.) and intraperitoneally (i.p.) with saline, 2) GM, treated s.c. with GM (70 mg/kg/12 hours/4 days), 3) AGE, treated i.p with AGE (1.2 mL/kg/12 hours/6 days), and 4) GM + AGE treated with GM and AGE. The treatment with AGE started two days before the first dose of GM (GM + AGE group) or saline (AGE group). Animals were sacrificed on day 5, and blood, urine, and kidneys were obtained. Nephrotoxicity was made evident by: 1) the increase in blood urea nitrogen and plasma creatinine, 2) the decrease in plasma glutathione peroxidase (GPx) activity and the urinary increase in N-acetyl-beta-D-glucosaminidase activity and total protein, and 3) necrosis of proximal tubular cells. These alterations were prevented or ameliorated by AGE treatment. Furthermore, AGE prevented the GM-induced increase in the renal levels of oxidative stress markers: nitrotyrosine and protein carbonyl groups and the decrease in manganese superoxide dismutase (Mn-SOD), GPx, and glutathione reductase (GR) activities. The protective effect of AGE was associated with the decrease in the oxidative stress and the preservation of Mn-SOD, GPx, and GR activities in renal cortex. These data suggest that AGE may be a useful agent for the prevention of GM-nephrotoxicity.

Physiological and morphological evidences for the presence acidocalcisomes in Trypanosoma evansi: single cell fluorescence and 31P NMR studies.

A new Ca(2+) intracellular store, the acidocalcisome, has been reported in trypanosomatids. It has been characterized physiologically as a Ca(2+) store sensitive to nigericin. The Ca(2+)/H(+)-ATPase is the system responsible for Ca(2+) accumulation, which depends on a pH gradient formed by ATP- and PPi-dependent proton pumps. In this work we present physiological and morphological evidences for the presence of acidocalcisomes in Trypanosoma evansi. The parasites were purified and loaded with the fluorescent dye Fura 2-AM in order to detect the intracellular changes of Ca(2+) levels in individual cells. The simultaneous incubation of T. evansi cells with ionomycin and nigericin led to large release of Ca(2+) (ca. 200 nM) from intracellular stores, which was not observed with either agent alone. On the other hand, no enhancement of the nigericin-induced Ca(2+) release was observed in the presence of oligomycin. Additionally, the pretreatment with bafilomycin decreases the nigericin-induced Ca(2+) release. These results confirm the presence of an intracellular non-mitochondrial acidic Ca(2+) storage compartment. These results suggest that H(+)-ATPase is involved in the process of Ca(2+) accumulation into the acidocalcisomes. Furthermore, the cells loaded with acridine orange exhibited abundant fluorescent vacuoles, which were sensitive to nigericin or bafilomycin A(1). Electronic transmission microscopy observations demonstrated the presence of electron dense particles in the parasites. High levels of inorganic pyrophosphate and triphosphate were detected in perchloric acid extracts of T. evansi by high resolution 31P NMR. Taken together, these results present the first evidence for the presence of acidocalcisomes in T. evansi.

[The emergence of multiple antimicrobial resistance in Vibrio cholerae isolated from gastroenteritis patients in Ceará, Brazil]. / Emergência da múltipla resistência a antimicrobianos em Vibrio cholerae isolados de pacientes com gastroenterite no Ceará, Brasil.

Of 7058 Vibrio cholerae strains recovered from patients suspected of cholera in the State of Ceará between December 1991 and September 1993, two were resistant to antimicrobials (Ampicillin, erythromycin, trimethoprim-sulfamethoxazole, tetracycline) and to vibriostatic agent O/129 (2,4-diamino-6,7-diisopropylpteridine). From the bacteriological standpoint, one strain was identified as V. cholerae serogroup O:1, biotype El Tor, serovar Inaba, and another as V. cholerae serogroup O:22, biochemically classified as Heiberg type II. It was shown that only in the serogroup O:1 strain, multiple resistance was encoded by a plasmid transferrable by conjugation to Escherichia coli K12 and a sensitive strains of V. cholerae O1 and non-O1, with at a frequency between 8 x 10(-2) and 5 x 10(-6). The plasmid, with a molecular weight of 147 Kb, encoded both multiple resistance to antimicrobials and the vibriostatic compound (O/129), compatible with descriptions reported in other parts of world.

Inactivation of chloramphenicol by O-phosphorylation. A novel resistance mechanism in Streptomyces venezuelae ISP5230, a chloramphenicol producer.

Plasmid pJV4, containing a 2.4-kilobase pair insert of genomic DNA from the chloramphenicol (Cm) producer Streptomyces venezuelae ISP5230, confers resistance when introduced by transformation into the Cm-sensitive host Streptomyces lividans M252 (Mosher, R. H. Ranade, N. P., Schrempf, H., and Vining, L. C. (1990) J. Gen. Microbiol. 136, 293-301). Transformants rapidly metabolized Cm to one major product, which was isolated and purified by reversed phase chromatography. The metabolite was identified by nuclear magnetic resonance spectroscopy and mass spectrometry as 3'-O-phospho-Cm, and was shown to have negligible inhibitory activity against Cm-sensitive Micrococcus luteus. The nucleotide sequence of the S. venezuelae DNA insert in pJV4 contains an open reading frame (ORF) that encodes a polypeptide (19 kDa) with a consensus motif at its NH2 terminus corresponding to a nucleotide-binding amino acid sequence (motif A or P-loop; Walker, J. E., Saraste, M., Runswick, M. J., and Gay, N. J. (1982) EMBO J. 1, 945-951). When a recombinant vector containing this ORF as a 1.6-kilobase pair SmaI-SmaI fragment was used to transform S. lividans M252, uniformly Cm-resistant transformants were obtained. A strain of S. lividans transformed by a vector in which the ORF had been disrupted by an internal deletion yielded clones that were unable to phosphorylate Cm, and exhibited normal susceptibility to the antibiotic. The results implicate the product of the ORF from S. venezuelae as an enzymic effector of Cm resistance in the producing organism by 3'-O-phosphorylation. We suggest the trivial name chloramphenicol 3'-O-phosphotransferase for the enzyme.

Indicación racional de la combinación de antibióticos con base en su sinergismo y antagonismo

La combinación de antibióticos en pediatría puede ser desde el punto de vista práctico, un tema de singular importancia dada la frecuencia con que los pacientes pediátricos sufren de patologías graves como consecuencia de su alta susceptibilidad, debida a estado inmunológico apenas en desarrollo, a sus condiciones nutricionales y al desarrollo tecnológico que coloca al recién nacido y al niño en condiciones de alto riesgo de ser colonizado por gérmenes. Sólo con una adecuada combinación de antimicrobianos podrán los pacientes pediátricos responder controlando la infección cuando se presenta. La combinación de antimicrobianos tiene indicaciones muy específicas: Ampliar el espectro terapéutico, producir sinergismo, limitar el desarrollo de resistencia y controlar infecciones polibacterianas. A su vez, la combinación de antibióticos conlleva ciertas limitaciones: Aumenta la toxicidad potencial de los antibióticos por separado, posibilita antagonismos por desconocimiento u omisión, aumenta los costos, crea una falsa seguridad en el médico. Por tal motivo es importante una guía racional sobre el manejo de la combinación de antibióticos, partiendo de los factores básicos que lo deben regir, conociendo con claridad los mecanismos de acción de los antibióticos comprometidos en la combinación, al igual que su farmacocinética y los aspectos relacionados con la población bacteriana sobre la que actúan. Sólo conociendo e integrando todos estos aspectos podremos hacer uso racional de la combinación de antibióticos.

Costs of antibiotic therapy

An increasing number of new and expensive antibiotics on the market makes it imperative that costs be contained and an attempt made to rationalise use without comprising patient care. Prerequisites of any containment programme are education of health and medical personnel and a good communication system to provide timely reports on culture and sensitivity results to personnel concerned