Arsenic reduces the GATA3 expression associated with an increase in proliferation and migration of mammary epithelial cell line MCF-10A.

Arsenic is associated with the development of breast cancer. However, the molecular mechanisms of arsenic induction of breast cancer are not fully defined. Interaction with zinc finger (ZnF) motifs in proteins is one of the proposed mechanisms of arsenic toxicity. GATA3 is a transcription factor that regulates the transcription of genes associated with cell proliferation, cell differentiation and the epithelial-mesenchymal transition (EMT) in mammary luminal cells. Given that GATA3 possesses two ZnF motifs essential for the function of this protein and that arsenic could alter the function of GATA3 through interaction with these structural motifs, we evaluated the effect of sodium arsenite (NaAsO2) on GATA3 function and its relevance in the development of arsenic-induced breast cancer. Breast cell lines derived from normal mammary epithelium (MCF-10A), hormone receptor-positive and hormone receptor negative breast cancer cells (T-47D and MDA-MB-453, respectively) were used. We observed a reduction on GATA3 protein levels at non-cytotoxic concentrations of NaAsO2 in MCF-10A and T-47D, but not in MDA-MB-453 cells. This reduction was associated with an increase in cell proliferation and cell migration in MCF-10A, but not in T-47D or MDA-MB-453 cells. The evaluation of cell proliferation and EMT markers indicate that the reduction on GATA3 protein levels by arsenic, disrupts the function of this transcription factor. Our data indicate that GATA3 is a tumor suppressor in the normal mammary epithelium and that arsenic could act as an initiator of breast cancer by disrupting the function of GATA3.

Daytime Restricted Feeding Modifies the Temporal Expression of CYP1A1 and Attenuated Damage Induced by Benzo[a]pyrene in Rat Liver When Administered before CYP1A1 Acrophase.

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that heterodimerizes with the AhR nuclear translocator (ARNT) to modulate CYP1A1 expression, a gene involved in the biotransformation of benzo[a]pyrene (BaP). The AhR pathway shows daily variations under the control of the circadian timing system. Daytime restricted feeding (DRF) entrains the expression of genes involved in the processing of nutrients and xenobiotics to food availability. Therefore, we evaluate if temporal AhR, ARNT, and CYP1A1 hepatic expression in rats are due to light/dark cycles or fasting/feeding cycles promoted by DRF. Our results show that AhR oscillates throughout the 24 h period in DRF and ad libitum feeding rats (ALF), showing maximum expression at the same time points. DRF modified the peak of ARNT expression at ZT5; meanwhile, ALF animals showed a peak of maximum expression at ZT17. An increased expression of CYP1A1 was linked to the meal time in both groups of animals. Although a high CYP1A1 expression has been previously associated with BaP genotoxicity, our results show that, compared with the ALF group, DRF attenuated the BaP-CYP1A1 induction potency, the liver DNA-BaP adducts, the liver concentration of unmetabolized BaP, and the blood aspartate aminotransferase and alanine aminotransferase activities when BaP is administered prior to the acrophase of CYP1A1 expression. These results demonstrate that DRF modifies the ARNT and CYP1A1 expression and protects from BaP toxicity.

Structure-Activity Relationship (SAR) and in vitro Predictions of Mutagenic and Carcinogenic Activities of Ixodicidal Ethyl-Carbamates.

Ethyl-4-bromophenyl-carbamate (LQM 919) and Ethyl-4-chlorophenyl-carbamate (LQM 996) are compounds that inhibit egg-laying and hatching of tick larvae that are resistant to conventional ixodicides. The structure-activity relationship (SAR) to get the endpoint predictions of mutagenicity and carcinogenicity of the LQM 919 and LQM 996 was performed and the absence of mutagenicity was confirmed by Ames test. SAR analysis show no structural alerts indicating the ability of ethyl-carbamates to bind biomolecules or estrogen receptors. Endpoint of mutagenicity with and without metabolic activation showed that the ethyl-carbamates were negative (p <0.05) for mutagenicity induction in strains TA97, TA98, TA102, TA1535, TA1537 and TA1538 of Salmonella typhimurium. Pre-incubation with different ethyl-carbamate concentrations did not increase the number of spontaneously reverting colonies; moreover, the compounds did not induce a concentration-dependent increase in the number of reverting colonies in any of the strains used. This confirmed the absence of mutagenic activity in this test system. Exogenous metabolic activation did not modify these observations; suggesting that no metabolites with mutagenic activity were present. The endpoint of carcinogenicity in rats were negative for LQM 919 (p <0.05,) and LQM 996 (p <0.001). The results of the present study strongly suggest that ethyl-carbamates do not represent a risk for cancer in mammals.

Differential inhibition of naringenin on human and rat cytochrome P450 2E1 activity.

Cytochrome P450 2E1 (CYP2E1) has been proposed as a molecular target in oxidative stress-associated metabolic diseases. Rats are chosen as model organisms in most experiments studying CYP2E1-related toxicity; however, the human relevance of these results remains unclear. To describe differences in catalysis and inhibition between human and rat CYP2E1, recombinant human and rat CYP2E1 enzymes were treated with different concentrations of naringenin (NAR, 10 nM - 1 mM), and inhibition parameters were calculated. Interspecies differences in the catalytic efficiency for O-demethylation of 7-methoxy-4-(trifluoromethyl)coumarin were revealed (45-fold higher in human CYP2E1 than in the rat enzyme). Additionally, differences in the potency of inhibition of NAR were found (absolute half inhibitory concentration, IC50 = 204 ± 28 and 69 ± 4 µM; inhibition constant, Ki = 9 ± 2 and 161 ± 20 µM in human and rat CYP2E1, respectively). Although NAR exhibited a noncompetitive mechanism of inhibition of both CYP2E1 enzymes, this compound is an irreversible inhibitor of rat CYP2E1 and a reversible inhibitor of the human enzyme. Molecular docking suggested that differences in the potency of inhibition and time dependence between species could be attributable to the differential interactions of NAR with access channels to the CYP2E1 catalytic site. These results highlight the importance of finding the appropriate model to improve the predictability of animal-based assays for human risk assessment.

Repetitive DNA profile of the amphibian mitogenome.

BACKGROUND: Repetitive DNA elements such as direct and inverted repeat sequences are present in every genome, playing numerous biological roles. In amphibians, the functions and effects of the repeat sequences have not been extensively explored. We consider that the data of mitochondrial genomes in the NCBI database are a valuable alternative to generate a better understanding of the molecular dynamic of the repeat sequences in the amphibians. RESULTS: This work presents the development of a strategy to identify and quantify the total amount of repeat sequences with lengths from 5 to 30 base pairs in the amphibian mitogenomes. The results show differences in the abundance of repeat sequences among amphibians and bias to specific genomic regions that are not easily associated with the classical amphibian ancestry. CONCLUSIONS: Derived from these analyses, we show that great variability of the repeat sequences exists among amphibians, demonstrating that the mitogenomes of these organisms are dynamic.

Human CYP1A1 inhibition by flavonoids.

Cytochrome P4501A1 (CYP1A1) is involved in the metabolism of several genotoxic/carcinogenic environmental xenobiotics including polycyclic aromatic hydrocarbons (PAHs) like benzo[a]pyrene. Several authors had proposed CYP1A inhibition as a plausible strategy for cancer chemoprevention. Using ethoxyresorufin O-deethylase activity (EROD), we tested the inhibitory properties of nine flavonoids: quercetin, miricetin, luteolin, fisetin, morin, kaempferol, 5-hydroxyflavone (5-HF), 3-hydroxyflavone (3-HF), and flavone (F) against human recombinant CYP1A1. The last three compounds exerted the highest inhibitory effect with IC50 values of 0.07, 0.10 and 0.08 µM respectively; the more hydroxyl-groups were present, the lower the potency of inhibition was. Biochemical characterization leads to the conclusion that flavone and its hydroxy derivatives are mixed-type inhibitors. In silico studies have shown that, Phe224 and other aromatic residues in the human CYP1A1 active site play an important role in flavonoid-CYP interaction, through a π/π stacking between the aminoacid and the flavonoid C-ring. Outside the active site, the three flavonoids bind preferentially between A and K helices of the enzyme. Results from the Ames test using human S9 fraction revealed that none of the three compounds was mutagenic. We can consider 5-HF, 3-HF, and F as potential chemopreventive agents against genotoxic damage caused by metabolites resulting from CYP1A1 activity.

Role of NF-κB in cytochrome P450 epoxygenases down-regulation during an inflammatory process in astrocytes.

Cytochrome P450 (CYP) epoxygenases and their metabolic products, epoxyeicosatrienoic acids (EETs), have been proposed as important therapeutic targets in the brain. However, CYP expression can be modified by the presence of diverse pro-inflammatory cytokines and the subsequent activation of the NF-κB pathway. It has been indicated that CYP epoxygenases are down-regulated by inflammation in the heart, kidney and liver. However, up to this point, there has been no evidence regarding regulation of CYP epoxygenases during inflammation in the brain. Therefore, in order to explore the effects of inflammation and NF-κB activation in CYP2J3 and CYP2C11 regulation, rat primary astrocytes cultures were treated with LPS with and without IMD-0354 (selective NF-κB inhibitor). Cyp2j3 and Cyp2c11 mRNA expression was determined by qRT-PCR; protein expression was determined by immunofluorescence and by Western Blot and total epoxygenase activity was determined by the quantification of EETs by ELISA. NF-κB binding sites in Cyp2j3 and Cyp2c11 promoter regions were bioinformatically predicted and Electrophoretic Mobility Shift Assays (EMSA) were performed to determine if each hypothetic response element was able to bind NF-κB complexes. Results shown that LPS treatment is able to down-regulate astrocyte CYP2J3 and CYP2C11 mRNA, protein and activity. Additionally, we have identified NK-κB as the transcription factor involved in this regulation.

Preclinical evidences of safety of a new synthetic adjuvant to formulate with the influenza human vaccine: absence of subchronic toxicity and mutagenicity.

Context: Influenza is a severe, life-threatening viral disease that can be prevented by vaccination. However, the anti-influenza human vaccine failed to show the required efficacy both in infants under 5 years old and in the elder population, who are among those with the highest risk of developing severe complications after influenza infection. Therefore, it is of high importance to improve the vaccine efficacy and ensure its safety in these susceptible populations. GK-1, a novel 18-aa peptide adjuvant, has been proved to increase the immunogenicity of the human influenza vaccine in both young and aged mice. Objective: A preclinical study of the toxicity profile of GK-1 following the World Health Organization guidelines to support its use was herein conducted. Material and methods: GK-1 was synthetically produced following Good Manufacturing Practices. The toxicological evaluation of GK-1 peptide was performed in rats after repeated dose-ranging trials by the subcutaneous route. The mutagenic potential of GK-1 was assessed by the micronucleus, chromosomal aberration, and Ames tests, in accordance with OECD Guidelines. Results: GK-1 did not show toxic effects at doses up to 12.5mg/kg, corresponding to 25 times the dose intended for human use. No indications of mutagenic potential were observed. GK-1 after dermal administration was well tolerated locally. Conclusion: The efficacy of GK-1 to improve influenza vaccine protection, along with the absence of toxicity and mutagenicity, as reported herein, support the evaluation of this peptide in a clinical trial as a novel adjuvant for human use.

Bacterial mutagenicity of selected procarcinogens in the presence of recombinant human or rat cytochrome P4501A1.

Cytochrome P4501A1 (CYP1A1) is an important enzyme of procarcinogen activation. We have studied bacterial (Ames test) mutagenicity resulting from mutagen activation by recombinant human or rat CYP1A1. Mutagenicity depends on both the chemical group and species-specific activation: polycyclic aromatic hydrocarbons showed higher (5-7-fold) mutagenic activity when activated by the human enzyme, whereas heterocyclic amines were more mutagenic (5-75-fold) in the presence of the rat enzyme. With regard to the two aromatic amines tested, only 2-aminoanthracene showed a clear species preference, activated 3-fold more effectively by human than by rat CYP1A1. We also analyzed in silico the binding of these compounds to the human and rat enzyme catalytic sites, identifying residues expected to participate in ligand recognition. A phenylalanine residue was involved in CYP-mutagen stabilization through π-π stacking. Variations in the three-dimensional conformations and distances to the heme groups may contribute to differences between human and rat CYP-substrate interactions. In conclusion, CYP1A1 shows significant differences between species, in terms of mutagen activation, which should be considered in the context of human risk assessment.

Cytochrome P450 in the central nervous system as a therapeutic target in neurodegenerative diseases.

Cytochromes P450 (CYPs) constitute a family of enzymes that can be found in the endoplasmic reticulum (ER), mitochondria or the cell surface of the cells. CYPs are characterized by carrying out the oxidation of organic compounds and they are mainly recognized as mediators of the biotransformation of xenobiotics to polar hydrophilic metabolites that can be eliminated from the organism. However, these enzymes play a key role in many other physiological processes, being involved in diverse indispensable metabolic pathways since they metabolize many endogenous substrates. Various CYP isoforms are expressed in the brain, and it is believed that this could be in part due to the particular function of brain CYPs. In the brain, CYPs are involved in the cholesterol turnover, the biosynthesis of dopamine, serotonin, morphine, hormones, and protective lipid mediators (epoxyeicosatrienoic acids), in addition to their already recognized role in xenobiotics detoxification and psychotropic drug metabolism. Increasing evidence suggests that this group of enzymes is fundamental for the normal functioning and maintenance of brain homeostasis. This review is focused on highlighting the importance of CYP-mediated endogenous metabolism in the central nervous system (CNS) and its relationship with recent findings regarding CYP involvement in neurodegenerative diseases. Some therapeutic approaches focused on CYP regulation are also discussed.

Proyecto URISCAM: Evaluación multicéntrica del citómetro UF-Series en el despistaje de infecciones urinarias / URISCAM project: multicenter evaluation of the UF-Series cytometer in the urinary tract infections screening

Introducción. El urocultivo, prueba de referencia para confirmar la existencia de Infección del tracto urinario (ITU), es la solicitud más demandada a Microbiología. Nuestro objetivo fue determinar la rentabilidad diagnóstica del citómetro UF-Series como método de cribado en la detección de ITU. Material y métodos. Se analizaron las orinas remitidas a los seis laboratorios de Microbiología participantes en un periodo de 5 días laborables. Se recogieron variables demográficas y de origen de la muestra, tipo de muestra (orina de media micción, sondaje, nefrostomía), recogida con/sin ácido bórico, lectura del citómetro (leucocituria, bacteriuria, morfología bacteriana y células epiteliales) y resultado del urocultivo. Para determinar la capacidad predictiva del citómetro se representaron las curvas ROC. Resultados. Se procesaron 2.468 muestras de pacientes con edad media de 53 años (ratio mujeres:hombres 2:1). El urocultivo detectó un 23% de orinas positivas. Las variables predictoras de ITU fueron: lectura morfológica de bacilos, bacteriuria ≥ 21 bacterias/μL, edad ≥ 65 años, procedencia de las muestras recogidas en urgencias y hospitalización, y presencia de conservante. Con el punto de corte de 21 bacterias, obtuvimos una sensibilidad del 93,3% y un VPN del 94,5%, lo que permitiría dejar de sembrar el 28,9% de las orinas recibidas con 1,6% de falsos negativos. Conclusiones. Consideramos que el UF-Series es una herramienta válida y precisa para la detección de ITU, por lo que podría utilizarse como cribado previo al urocultivo en la práctica clínica, reduciendo el número de orinas a sembrar en aproximadamente un 30% con una tasa baja de falsos negativos (AU)

Introduction. Urine culture, the gold standard to confirm the presence of urinary tract infection (UTI), is the most requested assay in the microbiology department. Our objective was to determine the diagnostic yield of the UF-Series cytometer as a screening method for UTI. Material and methods. All the urine samples sent to the six Microbiology Laboratories participating in a period of 5 working days were analyzed. We collected demographic variables, apart from those variables related to urine samples: source and sample type (midstream, catheterized or nephrostomy urines), collection with/without boric acid, cytometer parameters (leukocyturia, bacteriuria, bacteria morphology and epithelial cells) and urine culture results. ROC curves were plotted to determine predictive capacity of the cytometer. Results. A sample of 2,468 patients with average age of 53 years were processed (ratio women:men 2:1). Urine culture detected 23% of positive urine samples. The predictor variables of UTI were: morphology of bacilli, bacteriuria ≥21 bacteria/μL, age ≥65 years, samples collected in the emergency service and hospitalization and preserving conditions. With 21 bacteria/ μL as a cut-off point, we obtained a sensitivity of 93.3% and 94.5% negative predictive value, then reducing the samples to be cultured by 28.9% with 1.6% false negatives. Conclusions. We consider that the UF-Series is a valid and accurate tool for the detection of UTI. Therefore, it could be used as screening method in the clinical practice prior to the urine culture, reducing culture requirement by approximately 30%, with a low false negative rate (AU)

Inhibition of human and rat CYP1A1 enzyme by grapefruit juice compounds.

Cytochrome P4501A1 is involved in the metabolism of carcinogenic polycyclic aromatic hydrocarbons; therefore, its inhibition interferes with the carcinogenesis process induced by these compounds in rats. The human and rat CYP1A1 differ by 21% in amino acid sequence, including the active site of the enzyme; this difference may be an important factor when results obtained using animal models are interpolated to humans. Based on its previously reported CYP inhibitory properties, we studied the effects of two molecules contained within grapefruit juice, naringenin and 6',7'-dihydroxybergamottin, on human and rat CYP1A1 activity. For this purpose, the kinetics of inhibition as well as computational simulations were used. Naringenin and 6',7'-dihydroxybergamottin were found to be competitive inhibitors of human and rat CYP1A1. Additionally, naringenin exerted a mixed type inhibition effect on rat CYP1A1. Computational docking showed that inhibitors might block the oxidation of 7-ethoxyresorufin by binding to the CYP1A1 active site. Our results demonstrate the differences in CYP inhibitory mechanisms for the same molecule when CYP from different species are considered.

Utilidad de la espectrometría de masas en el diagnóstico microbiológico de la candiduria / Utility of mass spectrometry in the microbiological diagnosis of candiduria

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Regulation of Human Cytochrome P4501A1 (hCYP1A1): A Plausible Target for Chemoprevention?

Human cytochrome P450 1A1 (hCYP1A1) has been an object of study due to its role in precarcinogen metabolism; for this reason it is relevant to know more in depth the mechanisms that rule out its expression and activity, which make this enzyme a target for the development of novel chemiopreventive agents. The aim of this work is to review the origin, regulation, and structural and functional characteristics of CYP1A1 letting us understand its role in the bioactivation of precarcinogen and the consequences of its modulation in other physiological processes, as well as guide us in the study of this important protein.

A case-control study of post-traumatic endophthalmitis at a Spanish hospital.

Our aim was to identify the potential risk factors for developing post-traumatic endophthalmitis (PTE) and the possible measures of prevention. Retrospective case-control study, with 15 cases of PTE and 2 matched controls. We reviewed the medical records of the cases and their respective controls during the period 1996-2008 at a Spanish Hospital. We collected demographic data and information about the type of trauma, the potential risk factors, comorbidities, microbial isolations, antimicrobial susceptibility, administered treatments, and the visual outcome. The independent predictor factors identified for PTE were intraocular foreign body (IOFB) (OR 5.48; CI 95 % 1.05-28.7), dirty wound (OR 4.91; CI 95 % 0.96-25.3), and wound closure delays of 24 h or more (OR 5.48; CI 95 % 1.05-28.7). The probability of endophthalmitis in patients without these risk factors was 5.9 %, but ascended to 65.3 % and 90.3 %, in those patients with two and three risk factors, respectively. Infected patients presented a complication rate of 80 %, with an evisceration rate of 53 %; both were significantly associated with infection. The visual outcome was poor and related to the presence of IOFB and virulent microorganisms (Bacillus sp., filamentous fungus), visual acuity at presentation, and retinal detachment. Patients who presented an IOFB, dirty wound, and delayed wound closure were 15 times more likely to develop infection, and when infected, patients fared much worse than those non-infected. We thus recommend aggressive prophylactic measures in patients with these risk factors, adding antifungal prophylaxis when the injury is contaminated with vegetable matter.

Gas Chromatography-Mass Spectrometry Analysis of Ulva fasciata (Green Seaweed) Extract and Evaluation of Its Cytoprotective and Antigenotoxic Effects.

The chemical composition and biological properties of Ulva fasciata aqueous-ethanolic extract were examined. Five components were identified in one fraction prepared from the extract by gas chromatography-mass spectrometry, and palmitic acid and its ethyl ester accounted for 76% of the total identified components. Furthermore, we assessed the extract's antioxidant properties by using the DPPH, ABTS, and lipid peroxidation assays and found that the extract had a moderate scavenging effect. In an experiment involving preexposition and coexposition of the extract (1-500 µg/mL) and benzo[a]pyrene (BP), the extract was found to be nontoxic to C9 cells in culture and to inhibit the cytotoxicity induced by BP. As BP is biotransformed by CYP1A and CYP2B subfamilies, we explored the possible interaction of the extract with these enzymes. The extract (25-50 µg/mL) inhibited CYP1A1 activity in rat liver microsomes. Analysis of the inhibition kinetics revealed a mixed-type inhibitory effect on CYP1A1 supersome. The effects of the extract on BP-induced DNA damage and hepatic CYP activity in mice were also investigated. Micronuclei induction by BP and liver CYP1A1/2 activities significantly decreased in animals treated with the extract. The results suggest that Ulva fasciata aqueous-ethanolic extract inhibits BP bioactivation and it may be a potential chemopreventive agent.

Seroprevalence survey of zoonoses in Extremadura, southwestern Spain, 2002-2003.

Our aims were to determine the seroprevalence rates for the most common types of zoonosis among the population of Extremadura (southwestern Spain) and to identify the associated risk factors. We conducted a seroepidemiological survey to collect information on family background and the habits of people residing in Extremadura between 2002 and 2003. Antibodies to Brucella were determined by Rose Bengal staining and a standard tube agglutination test; a titer of 1/80 was considered to be positive. Antibody titers for spotted fever, leishmaniasis, echinococcosis, and toxoplasmosis were determined by enzyme-immunoassays. Independent risk factors identified were age (younger age for brucellosis), male gender (brucellosis, spotted fever, and toxoplasmosis), occupation and contact with animals (brucellosis and spotted fever for those in contact with goats, hydatidosis for those in contact with sheep, leishmaniasis for those in contact with dogs, and toxoplasmosis for those in contact with cats and pigs), and consuming contaminated food (brucellosis by eating fresh cheese, hydatidosis by eating homemade sausages, and toxoplasmosis by eating pork). Except for leishmaniasis, the other zoonoses were more prevalent in rural areas, and, with the exception of brucellosis, they were all more prevalent in Badajoz. The distribution of zoonoses in Extremadura was strongly influenced by keeping livestock and eating habits. Thus, brucellosis was more prevalent in Caceres (associated with cheese consumption), while toxoplasmosis (pork consumption) and spotted fever (from hunting) were more common in Badajoz.

[Trend in the susceptibility of the most frequent bacterial pathogens isolated at Hospital General La Mancha Centro over 2010-2012 period]. / Tendencia de sensibilidad de los patógenos bacterianos más frecuentemente aislados en el Hospital General La Mancha Centro durante el periodo 2010-2012.

Introduction. Our objective was to determine the trend of the antimicrobial susceptibility of the most common bacterial pathogens isolated in La Mancha Centro Hospital (MCH) between 2010-2012. Material and methods. Isolates of Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa from patients admitted to MCH were studied. These data and their antibiotic susceptibility were obtained from the database OBSERVA (BioMérieux). Results. The percentages of susceptibility for S. aureus were: 50% methicillin-resistant-S. aureus (MRSA) (higher co-resistance to erythromycin and levofloxacin), 46% erythromycin, 73% clindamycin, 45% levofloxacin, 99% rifampin and 100% cotrimoxazole, glycopeptides, linezolid and daptomycin. Increased resistance in ICU was observed (63% MRSA), with 50% of S. aureus (susceptible and methicillin-resistant strains) with vancomycin MIC values ≥ 0.5 mg/L. E. coli susceptibility: 62% amoxicillin-clavulanate, 55% ciprofloxacin, 60% cotrimoxazole, 84% gentamicin and 95% fosfomycin. K. pneumoniae susceptibility: 74% amoxicillin-clavulanate, 71% ciprofloxacin, 78% cotrimoxazole, 94% gentamicin and 87% fosfomycin. The percentage of BLEE strains was 17% and 21% for E. coli and K. pneumoniae, respectively, without detection of resistance to carbapenems. P. aeruginosa susceptibility: 80% ceftazidime and carbapenems, 63% ciprofloxacin and higher than 90% aminoglycosides. A decreasing trend of susceptibility to ceftazidime and carbapenems was observed in ICU and increasing trend to ciprofloxacin. Conclusions. Resistance percentages were higher in ICU than in the rest of the hospital, highlighting 63% of MRSA strains. Our percentage of BLEE and MRSA strains were higher than the Spanish media. Rifampicin and cotrimoxazole maintain good susceptibility to S. aureus, fosfomycin and aminoglycosides to Enterobacteriaceae and carbapenems to P. aeruginosa.