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Biomedical subjects

P Muñoz

Publications and source records attributed to P Muñoz.

At least 19 recordsLinked to original sources

Determination of chloramphenicol residues in shrimps by liquid chromatography-mass spectrometry.

A liquid chromatographic method with mass spectrometric detection and identification (LC-MS) is presented for the determination of chloramphenicol (CAP) in shrimp tissues. Homogenized shrimp samples were extracted with phosphate buffer (pH 7.0). Clean-up was carried out on a C(18) SPE cartridge. Chloramphenicol was determined by LC-MS-ESI in negative mode. The column used was a Symmetry Shield with a mixture of acetonitrile-water (25:75) as mobile phase. Shrimp samples were fortified at CAP levels between 0.2 and 50 ng g(-1) with 5D-CAP as internal standard. At these levels, accuracies lay between 101 and 110% and between-day reproducibilities were lower than 7.1%, expressed as the variation coefficient of the mean. Limit of decision (CCalpha) was 0.02 ng g(-1). Limit of quantitation (LOQ) was 0.2 ng g(-1).

Animals↗

A needleless closed system device (CLAVE) protects from intravascular catheter tip and hub colonization: a prospective randomized study.

Hub colonization and subsequent intraluminal progression due to frequent opening and manipulation of intravenous systems is the cause of many catheter-related infections (CRI). A prospective, comparative, randomized study was performed to assess a new closed-needleless hub device (CLAVE) compared with conventional open systems (COS). End-points were hub and skin colonization, catheter tip colonization, catheter-related bloodstream infection (CRBSI) and number of accidental needlesticks. All cultures were processed following standard semiquantitative microbiological techniques. The study involved patients who underwent heart surgery over an 11-month period in a post-surgical ICU. During the study period, 352 patients underwent major heart surgery and 1774 catheters were inserted. Overall, 865 catheters in 178 patients were allocated to the CLAVE system and 909 catheters in 174 patients to COS. The groups were similar regarding underlying conditions and risk factors for infection. Comparison of endpoint results in CLAVE and COS groups was as follows: incidence density per 1000 catheter-days of tip colonization: 59.2 versus 83.6 (P=0.003); of hub colonization: 7.56 versus 24.66 (P=0.0017); of skin colonization: 41.5 versus 58.9 (P=0.038); and of CRBSI 3.78 versus 5.89 (P=0.4). There was one accidental needlestick and one catheter-related prosthetic endocarditis in the COS group. Multivariate analysis showed that CLAVE use was an independent protective factor for tip colonization. CLAVE offered significant protection from catheter-tip and hub colonization.

Aged↗

Catheter-related infections: diagnosis and intravascular treatment.

The diagnosis of catheter-related infections relies on the presence of clinical manifestations of infection and the evidence of colonization of the catheter tip by bacteria, mycobacteria, or fungi. The reference method to confirm the latter requires the withdrawal of the catheter for culturing, which frequently turns out to be inconvenient, unnecessary and costly. New methods try to avoid these drawbacks and to assess the presence of tip colonization without withdrawal. Comparative quantitative blood cultures with a marked increase (> or = 5) in colony counts between blood obtained from the catheter lumen and from a peripheral vein simultaneously is one of those methods. It has a high sensitivity (>80%) and specificity (94-100%) but it is cumbersome and requires both an easy backflow of blood in the catheter and the existence of bacteremia. Cytocentrifugation and acridine orange staining of blood withdrawn from an infected catheter lumen has a sensitivity and a specificity of over 90% for the diagnosis of tip colonization. 'Superficial cultures' comprise the semiquantitative culture of the hub, of the skin surrounding the catheter entrance and of the first subcutaneous portion (1 cm) of the catheter after swabbing. The sensitivity of this method is >90%, specificity is >80%, and positive and negative predictive values for catheters (considering together those with and without clinical data of infection) are 66 and 97%, respectively. Endoluminal brushing has proved to be an impractical and unreliable procedure, at least in our experience. New methods based on the speed of bacterial growth to detectable levels of micro-organisms in conventional blood cultures are a new and interesting way of assessing catheter-related infections. Moreover, as the use of antimicrobial-coated catheters becomes more prevalent, the existing definitions of catheter colonization and catheter-related infection may need to be modified, because such coatings may lead to false-negative culture results. Many catheter infections, diagnosed without catheter withdrawal, can be handled nowadays with the so-called 'antibiotic lock-in technique', which consists in 'locking' the infected catheter lumen with a solution containing antibiotics. A high proportion of infected catheters, mainly those with coagulase-negative staphylococci, can be maintained in place and sterilized with this technique, including catheters in patients with therapeutic failure after receiving conventional intravenous antibiotic therapy. New diagnostic and therapeutic techniques may avoid the unnecessary withdrawal of thousands of efficient, difficult to replace and expensive intravascular lines.

Anti-Bacterial Agents↗

Demolition of a hospital building by controlled explosion: the impact on filamentous fungal load in internal and external air.

The demolition of a maternity building at our institution provided us with the opportunity to study the load of filamentous fungi in the air. External (nearby streets) and internal (within the hospital buildings) air was sampled with an automatic volumetric machine (MAS-100 Air Samplair) at least daily during the week before the demolition, at 10, 30, 60, 90,120, 180, 240, 420, 540 and 660 min post-demolition, daily during the week after the demolition and weekly during weeks 2, 3 and 4 after demolition. Samples were duplicated to analyse reproducibility. Three hundred and forty samples were obtained: 115 external air, 69 'non-protected' internal air and 156 protected internal air [high efficiency particulate air (HEPA) filtered air under positive pressure]. A significant increase in the colony count of filamentous fungi occurred after the demolition. Median colony counts of external air on demolition day were significantly higher than from internal air (70.2 cfu/m(3) vs 35.8 cfu/m(3)) (P < 0.001). Mechanical demolition on day +4 also produced a significant difference between external and internal air (74.5 cfu/m(3) vs 41.7 cfu/m(3)). The counts returned to baseline levels on day +11. Most areas with a protected air supply yielded no colonies before demolition day and remained negative on demolition day. The reproducibility of the count method was good (intra-assay variance: 2.4 cfu/m(3)). No episodes of invasive filamentous mycosis were detected during the three months following the demolition. Demolition work was associated with a significant increase in the fungal colony counts of hospital external and non-protected internal air. Effective protective measures may be taken to avoid the emergence of clinical infections.

Air Conditioning↗

Bloodstream infections after invasive nonsurgical cardiologic procedures.

OBJECTIVE: To define the incidence, risk factors, and characteristics of bloodstream infections (BSIs) after invasive nonsurgical cardiologic procedures (ICPs). METHODS: Retrospective case-control study; multivariate analysis. RESULTS: Between January 1991 and December 1998, 22 006 ICPs were performed in our hospital and 25 BSIs were documented within 72 hours after ICP. Overall incidence of bacteremia was 0.11% (25 cases) (0.24% after percutaneous transluminal coronary angioplasty [14 cases of 5625 patients], 0.06% [corrected] after diagnostic cardiac catheterization [9 cases of 14 034 patients], and 0.08% [corrected] after electrophysiologic studies [2 cases of 2347 patients]). These 25 patients with bacteremia were compared with 50 controls randomly selected among patients who underwent an ICP but did not have BSIs. Patient-related risk factors for BSI were age older than 60 years (20 cases [80%] vs 28 controls [56%]), valvular disease (4 [16%] vs 1 [2%]), congestive heart failure (7 [28%] vs 1 [2%]), indwelling bladder catheter before the ICP (5 [20%] vs 1 [2%]), more than 1 puncture for the ICP (5 [20%] vs 3 [6%]), a prolonged procedure (83.7 vs 65.1 minutes); and/or more than 1 ICP performed (2 [8%] vs 0). Multivariate analysis identified the presence of congestive heart failure (odds ratio, 21; 95% confidence interval, 6.8-66.0) and age older than 60 years (odds ratio, 1.9; 95% confidence interval, 1.9-6.3) as independent risk factors for BSI after ICP. Bloodstream infection was detected a median of 1.7 days after the procedure. Gram-negative bacteremia accounted for 17 cases (68%) of the BSIs. Among the patients with BSI, the duration of hospital stay was significantly increased (21 vs 6 days). The overall mortality rate was 0.009% for patients who underwent an ICP (8.0% for the 25 patients with bacteremia documented within 72 hours after ICP). CONCLUSIONS: Bloodstream infection should be included among the potential complications of ICP. Elderly patients with recent congestive heart failure episodes constitute a subgroup with a higher risk of postprocedure bacteremia. Therapy with antimicrobial agents against gram-positive and gram-negative bacteremia should be initiated after performing blood cultures in patients with signs suggestive of infection.

Adult↗

Management of urinary tract infections and lymphocele in renal transplant recipients.

The most frequent infectious complication after renal transplantation is urinary tract infection. This article deals with antimicrobial prophylaxis, treatment of early and relapsing urinary tract infections, and management of asymptomatic bacteriuria in renal transplant patients. The incidence of lymphocele after renal transplantation varies, and its treatment is still controversial. Management options are discussed.

Anti-Infective Agents, Urinary↗

Ku antigen is required to relieve G2 arrest caused by inhibition of DNA topoisomerase II activity by the bisdioxopiperazine ICRF-193.

Ku antigen is necessary for DNA double-strand break (DSB) repair through its ability to bind DNA ends with high affinity and to recruit the catalytic subunit of DNA-PK to the DSBs. Ku-deficient cells are hypersensitive to agents causing DSBs in DNA but also to the DNA topoisomerase II (topo II) inhibitor ICRF-193, which does not induce DSBs. This suggests a new role of Ku antigen, that is independent of DSB repair by DNA-PK. Here we characterize the basis for the hypersensitivity of Ku-deficient cells to ICRF-193. Chromosome condensation and segregation, which are dependent on topo II, but also the catalytic activity of topo II in late S-G2 were inhibited to a comparable extent when ICRF-193 was applied to Ku-deficient cells or wild-type cells. However, mutant cells arrested in G2 by ICRF-193 treatment were unable to progress into M phase upon drug removal, although drug-trapped topo II complexes were removed from DNA and the two isoforms of topo II recovered their catalytic activity as in wild-type cells. The reversibility of G2 arrest was recovered by complementation of mutant cells with a human Ku86 cDNA. Notably, chromosome condensation was abnormal in Ku-deficient cells after suppression of the G2 arrest by caffeine, even in the absence of ICRF-193. These results reflect the involvement of Ku-antigen in the cellular response to topo II inhibition, more particularly in relieving G2 arrest caused by topo II inhibition in late S/G2 and the subsequent recovery of chromosome condensation.

Animals↗

The impact of the introduction of the 14-3-3 protein assay in the surveillance of sporadic Creutzfeldt-Jakob disease in Catalonia.

The 14-3-3 assay in the CSF has a high predictive value for the diagnosis of sporadic Creutzfeldt-Jakob disease (CJD). Our objective was to analyse the trends in mortality and incidence rates of CJD in Catalonia, an autonomous community of Spain, setting the cut off point the date when the 14-3-3 assay was incorporated to our CJD surveillance system. Only definite and probable cases according to the criteria adapted from Masters and colleagues were included for the analysis. Familial cases were excluded after genetic analysis of the PrP gen. We found a quasi threefold increase in the incidence and mortality rates for CJD after introduction of the 14-3-3 assay in our surveillance system in 1997 as compared with the period 1993-1996. Our findings suggest an improvement of case ascertainment in part due to the introduction of the 14-3-3 assay in the surveillance system.

14-3-3 Proteins↗

Infections in systemic lupus erythematosus and rheumatoid arthritis.

Patients with systemic lupus erythematosus have a higher infection rate than the general population. It is estimated that at least 50% of them will suffer a severe infectious episode during the course of the disease. Improvements in the control of the disease are discussed in this article.

Arthritis, Rheumatoid↗

A European perspective on nosocomial urinary tract infections II. Report on incidence, clinical characteristics and outcome (ESGNI-004 study). European Study Group on Nosocomial Infection.

OBJECTIVES: To estimate the incidence of nosocomially acquired urinary tract infections (NAUTI) in Europe and provide information on the clinical characteristics, underlying conditions, etiology, management and outcome of patients. MATERIALS AND METHODS: We collected clinical information from NAUTI patients with a microbiology report on the named study day. RESULTS: A total of 141 hospitals from 25 European countries participated in the study. Written institutional bladder catheter guidelines were in place in 90.3% of EU hospitals and 55% of non-EU hospitals (P < 0.05). The total number of new NAUTI episodes on the day of the study was 298, representing an incidence of 3.55 episodes/1000 patient-days and an estimated prevalence of 10.65/1000. The five most commonly isolated micro-organisms were Escherichia coli, Enterococcus sp., Candida sp., Klebsiella sp. and Pseudomonas aeruginosa. Patients from non-EU countries were younger, with more severe underlying diseases with a higher incidence of obstructive uropathy/lithiasis. Overall, 22.8% of patients had no 'classic' UTI-predisposing factors. Catheter-associated UTI (CAUTI) was present in 187 patients (62.8%). A closed drainage system was used in only 78.5% of catheterised patients. The indication for bladder catheterisation was not considered adequate in 7.6% of cases and continuation of bladder catheterisation was considered unnecessary in 31.3%. Opening of the closed drainage system was the most frequent major error in catheter management (16.8%). Antimicrobial treatment was not considered adequate in 19.8% of all cases. CONCLUSIONS: The incidence of NAUTI in a large European population is 3.55/1000 patient-days. There is clearly room for improvement in the area of bladder catheterisation, catheter care and medical management of NAUTI. We recommend that European authorities draw up and implement practical and specific guidelines to reduce the incidence of this infection.

Age Factors↗

A European perspective on nosocomial urinary tract infections I. Report on the microbiology workload, etiology and antimicrobial susceptibility (ESGNI-003 study). European Study Group on Nosocomial Infections.

OBJECTIVES: To obtain information on the microbiology workload, etiology and antimicrobial susceptibility of urinary tract infection (UTI) pathogens isolated in European hospitals. MATERIALS AND METHODS: We collected data available in the microbiology units of a large sample of European hospitals regarding the laboratory workload, diagnostic criteria, and etiology and antimicrobial resistance of the urinary isolates collected on one day (the study day). RESULTS: Data were received from a total of 228 hospitals from 29 European countries. The average rate of urine samples cultured per 1000 admissions in 1999 was 324. The criteria to consider a positive urine culture as significant were quite variable; > or =10(4) colony-forming units (CFU)/mL for bacteria or > or =10(3) CFU/mL in the case of yeasts were the most used cut-off points. On the study day, a total of 607 micro-organisms from 522 patients with nosocomial UTI were isolated. The six most commonly isolated micro-organisms were, in decreasing order: Escherichia coli (35.6%), Enterococci (15.8%), Candida (9.4%), Klebsiella (8.3%), Proteus (7.9%) and Pseudomonas aeruginosa (6.9%). Pseudomonas was isolated more frequently in non-EU countries. The study data reveal high rates of antimicrobial resistance in UTI pathogens, especially in non-EU countries, where Pseudomonas aeruginosa presented rates of aminoglycoside resistance as high as 72% to gentamicin, 69.2% to tobramycin and 40% to amikacin. CONCLUSIONS: Nosocomial UTI accounts for an important proportion of the workload in microbiology laboratories. A consensus on the practice and interpretation of urine cultures in Europe is needed. The levels and patterns of resistance of UTI pathogens must be a serious cause for concern and a clear reason for stricter guidelines and regulations in antimicrobial policy.

Anti-Bacterial Agents↗

Linezolid: pharmacokinetic characteristics and clinical studies.

Linezolid is an oxazolidinone indicated in the treatment of nosocomial and community-acquired pneumonia, complicated and uncomplicated skin and skin structure infections and vancomycin-resistant Enterococcus infections. The drug is also approved for use in complicated skin infections and nosocomial pneumonia caused by methicillin-resistant Staphylococcus aureus, concurrent bacteremia associated with vancomycin-resistant Enterococcus faecium and concurrent bacteremia associated with community-acquired pneumonia caused by penicillin-susceptible Streptococcus pneumoniae.

Acetamides↗

"Second-look" cytotoxicity: an evaluation of culture plus cytotoxin assay of Clostridium difficile isolates in the laboratory diagnosis of CDAD.

Clostridium difficile is one of the most frequent causes of hospital-acquired diarrhoea. Our objective was to prove that some stool samples with a direct negative cytotoxicity assay may indeed harbour toxigenic C. difficile and that this can be demonstrated by performing a "second-look" cytotoxicity assay using the isolated C. difficile strains. Over an eight-year period (1992-1999), the 8241 stool samples submitted for direct cell culture from patients with suspected C. difficile-associated diarrhoea (CDAD) were simultaneously plated on cycloserine cefoxitin fructose agar. C. difficile strains isolated from samples with a negative direct cell culture assay were re-tested for toxin production "second-look" cell culture assay). Using both methods 6423 samples (78%) were negative. Of the remaining 1818 samples, 127 (7%) yielded C. difficile isolates which were confirmed as non-producers of toxin by both methods, 1437 (85%) were positive in direct cell culture assay, and 254 were positive only after the "second-look" cell culture assay. Thus, our approach allowed us to detect an extra 15% of toxin-producing strains that could have gone undetected otherwise.The combination of direct-cell culture assay, culture for toxigenic C. difficile and "second-look" cell culture assay enhances the potential for diagnosis of CDAD and enables us to be more efficient with our patient care resources.

Bacterial Toxins↗

In vitro activity of new quinolones against Clostridium difficile.

We evaluated the in vitro activities of ofloxacin, levofloxacin, grepafloxacin, trovafloxacin and ciprofloxacin against Clostridium difficile. The MIC(90) was 128 mg/L for ofloxacin and levofloxacin, 64 mg/L for ciprofloxacin, 16 mg/L for grepafloxacin and 8 mg/L for trovafloxacin. Thirty per cent of isolates were resistant to trovafloxacin, and rates of resistance to ofloxacin, levofloxacin, grepafloxacin and ciprofloxacin were considerably higher. None of the antimicrobials studied would be a reliable therapeutic option against C. difficile. Whether some of the new fluoroquinolones can induce C. difficile-associated diarrhoea remains to be answered.

4-Quinolones↗

Dormancy, ABA content and sensitivity of a barley mutant to ABA application during seed development and after ripening.

Assessment of dormancy inception, maintenance and release was studied for artificially dried immature seeds harvested throughout seed development in the barley cv. Triumph and its mutant line TL43. Each was grown in Spain and Scotland under low and high dormancy inducing conditions, respectively. Both TL43 and Triumph followed a similar pattern of release from dormancy across the seasons, although seeds of TL43 were able to germinate at an earlier seed development stage. Abscisic acid (ABA) content was also studied in immature grains throughout the seed development period. Total amount of ABA in seeds of Triumph and TL43 was higher in plants grown in Scotland than in Spain. However, no clear genotypic differences in ABA pattern in the course of grain development could be detected whilst significant genotypic differences were observed for germination percentage (GP). Endogenous ABA content alone throughout grain development did not explain genetic differences in GP within environments. Environmental and genetic differences in dormancy were also observed on mature seeds throughout the after-ripening period. The initial germination (GP(0)) played a key role in the sensitivity to ABA of post-harvest mature seeds. For the same after-ripening stage, TL43 was more insensitive to exogenous ABA than Triumph. However, ABA responses in seeds of the two genotypes with similar GP(0) at different after-ripening stages were comparable. Therefore, differences in exogenous ABA sensitivity of post-harvest mature grain of these two genotypes seemed to be determined by, or coincident with, the initial germination percentage.

Abscisic Acid↗