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Alvaro Pascual

Publications and source records attributed to Alvaro Pascual.

At least 37 records · Page 2Linked to original sources
PubMed2005

Efficacy of cefepime and imipenem in experimental murine pneumonia caused by porin-deficient Klebsiella pneumoniae producing CMY-2 beta-Lactamase.

Previous studies have shown decreased in vitro activity of zwitterionic cephalosporins and carbapenems against porin-deficient Klebsiella pneumoniae expressing a plasmid-mediated AmpC-type beta-lactamase (PACBL). The in vitro and in vivo activities of cefepime and imipenem were evaluated against the porin-deficient strain K. pneumoniae C2 and its CMY-2-producing derivative [K. pneumoniae C2(pMG248)]. The MICs (in micrograms/milliliter) of cefepime and imipenem against K. pneumoniae C2 were 0.125 and 0.25, respectively, while the corresponding values against K. pneumoniae C2(pMG248) were 8 and 16. Cefepime showed a greater inoculum effect than imipenem against both strains. Imipenem showed a significant postantibiotic effect (>2 h) against K. pneumoniae C2(pMG248) at 1x, 2x, 4x, 6x, and 8x MIC. The maximum concentrations of drug in serum of cefepime and imipenem in a pneumonia model using mice were 124.1 and 16.9 mug/ml, respectively. DeltaT/MIC for K. pneumoniae C2 and C2(pMG248) were 1.29 h and 0.34 h for imipenem and 2.96 h and 1.27 h for cefepime. Both imipenem (30 mg/kg of body weight every 3 h) and cefepime (60 mg/kg every 4 h), administered for 72 h, increased the survival rate (86.6% and 100%) compared with untreated control animals (26.6%, P < 0.003) infected with K. pneumoniae C2. For the CMY-2-producing strain, imipenem, but not cefepime, increased the survival rate compared to the controls (86.6% and 40% versus 40%, P < 0.01). Bacterial concentration of the lungs was significantly decreased by both antimicrobials. In conclusion, imipenem was more active in terms of survival than cefepime for the treatment of murine pneumonia caused by a porin-deficient K. pneumoniae expressing PACBL CMY-2.

Animals↗
PubMed2004

[Clonal diversity and antimicrobial susceptibility of Acinetobacter baumannii isolated in Spain. A nationwide multicenter study: GEIH-Ab project (2000)].

INTRODUCTION: A nationwide multicenter study was performed in Spain to evaluate the clonal diversity and antimicrobial susceptibility of Acinetobacter baumannii clinical isolates. METHODS: A total of 221 consecutive A. baumannii isolates recovered from clinical samples from 25 Spanish hospitals during November 2000 were studied. Isolate identification was performed by phenotyping methods and by amplified rDNA restriction analysis. Clonal relationships among A. baumannii isolates were determined by pulsed field gel electrophoresis. MICs of amikacin (AK), ampicillin (AP), cephalothin (CF), cefoxitin (FX), ceftazidime (CZ), ciprofloxacin (CP), cotrimoxazole (T/S), doxycycline (DX), gemifloxacin (GX), gentamicin (GN), imipenem (IP), meropenem (MP), minocycline (MI), piperacillin (PP), polymyxin B (PB), rifampicin (RI), tetracycline (TT), sulbactam (SB) and tobramycin (TO) were determined by microdilution (NCCLS guidelines). RESULTS: Seventy-nine A. baumannii clones were differentiated. MIC50/MIC90 (mg/L) values for the 221 A. baumannii isolates were PP: > 512/> 512; AP, CF, FX: > 256/> 256; TT, GN: > 128/> 128; CZ: 128/> 256; CP: > 64/> 64; FP: 64/256; AK: 32/256; DX: 32/64; GX: > 16/> 16; TO: 16/128; SB, T/S: 16/64; MP: 8/> 128; IP: 4/128; RF: 4/8; MI: 2/16 and PB: 1/2. Percentages of susceptible isolates were PB: 100%; MI: 65.8%; IP: 52.5%, RF: 49.3%; SB: 46.7%; MP: 43.1%; AK: 34.7%; DX: 32.0%; TO: 21.3% and CZ, FP, GN, T/S, TT, GX, CP, AP, PP, CF and FX: < 20%. CONCLUSIONS: A. baumannii isolates show high clonal variability in Spain. The most active antimicrobial agents against this organism were polymyxin B, minocycline, rifampicin, imipenem, sulbactam, meropenem, amikacin and doxycycline.

Acinetobacter Infections↗
PubMed2004

Clinical features and epidemiology of Acinetobacter baumannii colonization and infection in Spanish hospitals.

OBJECTIVE: To investigate the clinical features and the epidemiology of Acinetobacter baumannii in Spanish hospitals. DESIGN: Prospective multicenter cohort study. SETTING: Twenty-seven general hospitals and one paraplegic center in Spain. METHODS: All cases of A. baumannii colonization or infection detected by clinical samples during November 2000 were included. Isolates were identified using phenotypic and genotypic methods. The molecular relatedness of the isolates was assessed by pulsed-field gel electrophoresis. RESULTS: Twenty-five (89%) of the hospitals had 221 cases (pooled rate in general hospitals, 0.39 case per 1,000 patient-days; range, 0 to 1.17). The rate was highest in intensive care units (ICUs). Only 3 cases were pediatric. The mean age of the patients in the general hospitals was 63 years; 69% had a chronic underlying disease and 80% had previously received antimicrobial treatment. Fifty-three percent of the patients had an infection (respiratory tract, 51%; surgical site, 16%; and urinary tract, 11%). Crude mortality was higher in infected than in colonized patients (27% vs 10%; relative risk, 1.56; 95% confidence interval, 1.2 to 2.0; P = .003). Molecular analysis disclosed 79 different clones. In most hospitals, a predominant epidemic clone coexisted with other sporadic clones. Imipenem resistance was present in 39% of the hospitals. CONCLUSIONS: A. baumannii was present in most participating Spanish hospitals (particularly in ICUs) with different rates among them. The organisms mainly affected predisposed patients; half of them were only colonized. Epidemic and sporadic clones coexisted in many centers.

Acinetobacter Infections↗
PubMed2004

Epidemiology and clinical features of infections caused by extended-spectrum beta-lactamase-producing Escherichia coli in nonhospitalized patients.

Infections due to extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESBLEC) in nonhospitalized patients seem to be emerging in different countries. Their incidence, epidemiology, and clinical impact in the community have not been studied. We describe the epidemiology and clinical features of infections caused by ESBLEC in nonhospitalized patients in Spain and the results of a case-control study performed to investigate the risk factors associated with the acquisition of these organisms. The clonal relatedness of the organisms was assessed by repetitive extragenic palindromic sequence PCR. The ESBLs and the genes encoding the ESBLs were initially characterized by isoelectric focusing and PCR, respectively. Forty-nine patients (76% with urinary tract infections, 22% with asymptomatic bacteriuria, and 2% with acute cholangitis) were included. Six patients were bacteremic. Diabetes mellitus (odds ratio, 5.5; 95% confidence interval, 1.6 to 18.7), previous fluoroquinolone use (odds ratio, 7.6; 95% confidence interval, 1.9 to 30.1), recurrent urinary tract infections (odds ratio, 4.5; 95% confidence interval, 1.3 to 15.1), a previous hospital admission (odds ratio, 18.2; 95% confidence interval, 5.3 to 61.1), and older age in male patients (odds ratio per year, 1.03; 95% confidence interval, 1.03 to 1.05) were identified as risk factors by multivariate analysis. The ESBLEC isolates were not clonally related. The ESBLs were characterized as members of the CTX-M-9 group, the SHV group, and the TEM group in 64, 18, and 18% of the isolates, respectively. ESBLEC is an emergent cause of urinary tract infections in nonhospitalized patients. There was no evidence of horizontal transmission of ESBLEC strains. Avoidance of fluoroquinolone use in high-risk patients should be considered whenever possible in order to avoid the selection of these organisms.

Adult↗
PubMed2004

Use of positive blood cultures for direct identification and susceptibility testing with the vitek 2 system.

In order to further decrease the time lapse between initial inoculation of blood culture media and the reporting of results of identification and antimicrobial susceptibility tests for microorganisms causing bacteremia, we performed a prospective study in which specially processed fluid from positive blood culture bottles from Bactec 9240 (Becton Dickinson, Cockeysville, Md.) containing aerobic media were directly inoculated into Vitek 2 system cards (bio-Mérieux, France). Organism identification and susceptibility results were compared with those obtained from cards inoculated with a standardized bacterial suspension obtained following subculture to agar; 100 consecutive positive monomicrobic blood cultures, consisting of 50 gram-negative rods and 50 gram-positive cocci, were included in the study. For gram-negative organisms, 31 of the 50 (62%) showed complete agreement with the standard method for species identification, while none of the 50 gram-positive cocci were correctly identified by the direct method. For gram-negative rods, there were 50% categorical agreements between the direct and standard methods for all drugs tested. The very major error rate was 2.4%, and the major error rate was 0.6%. The overall error rate for gram-negatives was 6.6%. Complete agreement in clinical categories of all antimicrobial agents evaluated was obtained for 19 of 50 (38%) gram-positive cocci evaluated; the overall error rate was 8.4%, with 2.8% minor errors, 2.4% major errors, and 3.2% very major errors. These findings suggest that the Vitek 2 cards inoculated directly from positive Bactec 9240 bottles do not provide acceptable bacterial identification or susceptibility testing in comparison with corresponding cards tested by a standard method.

Anti-Bacterial Agents↗
PubMed2003

Evaluation of SDS-polyacrylamide gel systems for the study of outer membrane protein profiles of clinical strains of Acinetobacter baumannii.

The outer membrane protein (OMP) profiles of 23 blood isolates of Acinetobacter baumannii representing all the different antimicrobial susceptibility patterns observed during a 3-year period in a Spanish hospital were studied. OMPs extracted from envelopes of sonicated cells after solubilisation with 2% of N-lauryl-sarcosinate were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Laemmli's buffers. Eight running gel systems differing in the concentration of polyacrylamide (8%, 10% and 12%) and in the absence or presence of urea (4 M and 6 M) were used in a preliminary study analysing the OMP profiles of four clonally unrelated strains of A. baumannii. When this study was completed, the OMPs of the 23 A. baumannii were analysed in 10% SDS-polyacrylamide gels with 6 M urea and in 12% SDS-polyacrylamide gels. Ten OMP profiles were observed in 10% SDS-polyacrylamide gels with 6 M urea, whereas only 5 OMP profiles were visualised using 12% SDS-polyacrylamide gels. The OMP profiles obtained in 10% SDS-polyacrylamide gels with 6 M urea only partially correlated with those observed in 12% SDS-polyacrylamide gels. In conclusion, the use of 10% SDS-polyacrylamide gels with 6 M urea is recommended for the study of OMP profiles of A. baumannii.

Acinetobacter Infections↗
PubMed2003

[Acinetobacter baumannii bacteremia: clinical and prognostic features].

OBJECTIVE: The incidence, clinical features, and prognosis of bacteremia due to Acinetobacter baumannii were investigated. METHODS: Prospective study of all episodes of A. baumanii bacteremia detected during the period of January 1995 to December 1998. A. baumannii was identified using recent standard methods. RESULTS: A total of 133 episodes of bacteremia due to A. baumannii were studied, all of them nosocomial-acquired. The incidence-density diminished from 2.02 episodes per 10,000 patient-days to 0.40 episodes per 10,000 patient-days after the implementation of a control program. Most of the patients (70%) were, or had been, in the ICU when bacteremia occurred. Some 80% of patients had a chronic illness and 62% had a Hilf's severity score > 4. Among the strains identified, 74% were multidrug-resistant and 28% were imipenem-resistant. Attributable mortality was 25.6%. Multivariate analysis showed that inappropriate antibiotic treatment, septic shock, and high Hilf's severity score were associated with poorer prognosis. CONCLUSION: A. baumannii bacteremia mainly affects severely ill patients who have undergone several invasive procedures, and who may have relevant associated morbidity and mortality. Among other variables, inappropriate antibiotic treatment was a risk factor for increased mortalilty.

Acinetobacter Infections↗
PubMed2003

[Comparative in vitro activity of garenoxacin (BMS-284756). Sentry program, Spain (1999-2000)].

INTRODUCTION: To evaluate the in vitro activity of the new des-fluoro quinolone, garenoxacin (BMS-284756), compared to activities of ciprofloxacin, levofloxacin, and gatifloxacin in clinical isolates recovered over 1999 and 2000 within the SENTRY antimicrobial surveillance program. METHODS: Quinolone-MICs were performed using the standard NCCLS microdilution technique in 2599 isolates recovered from Hospital Ramón y Cajal (Madrid), Virgen Macarena (Sevilla), and Bellvitge (Barcelona). RESULTS: The modal MIC range value exhibited by garenoxacin ( < or = 0.03-0.12 mg/L) for Enterobacteriaceae was similar to that of the other quinolones tested. A total of 70% of Pseudomonas aeruginosa isolates were susceptible to garenoxacin and 85% to ciprofloxacin and levofloxacin. Garenoxacin exhibited the highest activity in Staphylococcus aureus, including both methicillin-susceptible and -resistant isolates, with MIC90 values of < or = 0.03 and 2 mg/L, respectively. All Streptococcus pneumoniae isolates were susceptible to garenoxacin, regardless of their penicillin susceptibility status; in terms of MIC90, garenoxacin was 16 times more active than ciprofloxacin and levofloxacin and 4-8 times more active than gatifloxacin. All 6 ciprofloxacin-resistant S. pneumoniae strains showed garenoxacin MIC values ranging from < or = 0.03 to 0.5 mg/L. In Haemophilus influenzae and Moraxella catarrhalis, garenoxacin displayed excellent in vitro activity (MIC < or = 0.06 mg/L), similar to that of the other quinolones tested. CONCLUSIONS: Garenoxacin activity was similar to the activity of other quinolones in Enterobacteriaceae, but was lower in P. aeruginosa. Garenoxacin activity was clearly higher than that of other quinolones in gram-positive isolates, including methicillin-resistant S. aureus and S. pneumoniae with reduced penicillin susceptibility.

Anti-Infective Agents↗
PubMed2003

Effect of siliconized latex urinary catheters on the activity of carbapenems against Pseudomonas aeruginosa strains with defined mutations in ampC, oprD, and genes coding for efflux systems.

The decreased activity of carbapenems against Pseudomonas aeruginosa in the presence of eluates from siliconized latex urinary catheters is related to the loss of OprD and the expression of a new outer membrane protein (OMP). To understand this phenomenon, the activity of carbapenems and quinolones against P. aeruginosa strains, some producing AmpC beta-lactamase (BL), OprD, MexA-MexB-OprM and MexE-MexF-OprN, were determined in Mueller-Hinton broth (MH) and in MH-containing eluates. OMP profiles and BL activities were also studied. Resistance to imipenem in P. aeruginosa grown in eluate is due to both the loss of OprD and expression of BL. For meropenem this phenomenon is related to both the loss of OprD and the integrity of MexA-MexB-OprM.

Anti-Infective Agents↗
PubMed2003

Characterization and selection of HIV-1 subtype C isolates for use in vaccine development.

HIV-1 genetic diversity among circulating strains presents a major challenge for HIV-1 vaccine development, particularly for developing countries where less sequence information is available. To identify representative viruses for inclusion in candidate vaccines targeted for South Africa, we applied an efficient sequence survey strategy to samples from recently and chronically infected persons residing in potential vaccine trial sites. All 111 sequences were subtype C, including 30 partial gag, 26 partial pol, 27 V2-V3 env, and 28 V5-partial gp41 sequences. Of the 10 viruses cultured from recently infected individuals, 9 were R5 and 1 was R5X4. Two isolates, Du151 and Du422, collected within 2 months of infection, were selected as vaccine strains on the basis of their amino acid similarity to a derived South African consensus sequence The selection of recently transmitted R5 isolates for vaccine design may provide an advantage in a subtype C R5-dominant epidemic. The full-length Du422 gag and Du151 pol and env genes were cloned into the Venezuelan equine encephalitis (VEE) replicon particle (VRP) expression system. Du422 Gag protein expressed from the VRP accumulated to a high level and was immunogenic as demonstrated by cytotoxic T lymphocyte responses in mice vaccinated with gag-VRPs. Optimization of codon use for VRP expression in human cells did not enhance expression of the gag gene. The cloned Du151 env gene encoded a functional protein as demonstrated by fusion of VRP-infected cells with cells expressing CD4 and CCR5. Genes identified in this study have been incorporated into the VEE VRP candidate vaccines targeted for clinical trial in South Africa.

AIDS Vaccines↗
PubMed2003

Relationship between beta-lactamase production, outer membrane protein and penicillin-binding protein profiles on the activity of carbapenems against clinical isolates of Acinetobacter baumannii.

Twenty blood isolates of Acinetobacter baumannii were studied, representing eight pulsed-field gel electrophoresis patterns and all different antimicrobial susceptibility patterns observed during 1995-97 at the University Hospital Virgen Macarena, Seville, Spain. The MIC(90)s (mg/L) of imipenem and meropenem decreased from 16 to 0.5 and from 8 to 4, respectively, in the presence of BRL 42715 (BRL) but not clavulanic acid. Hydrolysing activity (nmol/min/mg) of bacterial supernatants against cefaloridine ranged from 8.8 to 552.3 for A. baumannii type I (imipenem MICs < or = 2), which expressed only a beta-lactamase of pI > or = 9, and from 12.3 to 1543.5 for A. baumannii type II (imipenem MICs > or = 4), which expressed a beta-lactamase of pI > or = 9 and two others of pI 6.3 and 7. The hydrolysing activities of A. baumannii type II against imipenem, meropenem and oxacillin were higher than those observed for A. baumannii type I. Ten outer membrane protein (OMP) profiles (A. baumannii types I and II) were visualized on 10% SDS-PAGE gels with 6 M urea, whereas only five OMP profiles (A. baumannii types I and II) were differentiated in 12% SDS-PAGE gels. Five A. baumannii with OMP profile type B, characterized by the absence of a 22.5 kDa OMP, were resistant to meropenem and/or imipenem. Twelve penicillin-binding protein (PBP) patterns were observed. PBP patterns of A. baumannii type II were characterized by the absence of a 73.2 kDa band (PBP 2). We concluded that production of beta-lactamases of pI 6.3 and 7.0 and reduced expression of PBP 2 are the most frequently observed mechanisms of resistance to carbapenems. In some isolates, loss of a 22.5 kDa OMP is also related to resistance to carbapenems.

Acinetobacter baumannii↗
PubMed2003

Role of Klebsiella pneumoniae OmpK35 porin in antimicrobial resistance.

OmpK35 from Klebsiella pneumoniae is the homologue of Escherichia coli OmpF porin. Expression of OmpK35 in K. pneumoniae strain CSUB10R (lacking both OmpK35 and OmpK36) decreased the MICs of cephalosporins and meropenem > or = 128-fold and decreased the MICs of imipenem, ciprofloxacin, and chloramphenicol > or = 8-fold. MIC reductions by OmpK35 were 4 times (cefepime), 8 times (cefotetan, cefotaxime, and cefpirome), or 128 times (ceftazidime) higher than those caused by OmpK36, but the MICs were similar or 1 dilution lower for other evaluated agents.

Amino Acid Sequence↗
PubMed2003

Activities of ABT-773 against Listeria monocytogenes and coryneform bacteria of clinical interest.

The in vitro activities of ABT-773 were evaluated against 15 Listeria monocytogenes strains and 196 coryneform bacteria isolated from clinical samples. One hundred percent of the L. monocytogenes strains were inhibited by </=0.015 micro g of ABT-773/ml. MICs of ABT-773 ( micro g/ml) at which 50% of the isolates tested were inhibited (MIC(50)s) and MIC(90)s for other organisms were 0.125 and 0.5 (Corynebacterium amycolatum), 1 and >32 (Corynebacterium jeikeium), 0.03 and >32 (Corynebacterium minutissimum), >32 and >32 (Corynebacterium pseudodiphtheriticum and Corynebacterium urealyticum), 0.125 and >32 (Corynebacterium striatum), and 0.03 and 0.5 (Rhodococcus equi), respectively.

Anti-Bacterial Agents↗