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1.
Epidemiol Infect ; 148: e149, 2020 05 26.
Article in English | MEDLINE | ID: mdl-32450935

ABSTRACT

Persistent methicillin-resistant Staphylococcus aureus (MRSA) infection in cystic fibrosis (CF) patients has been associated with a more rapid decline in lung function, increased hospitalisation and mortality. The aim of this study was to evaluate the clonal relationships among 116 MRSA isolates from 12 chronically colonised CF pediatric patients over a 6-year period in a Rio de Janeiro CF specialist centre. Isolates were characterised by antimicrobial resistance, SCCmec type, presence of Panton-Valentine Leukocidin (PVL) genes and grouped according to DNA macrorestriction profile by pulsed-field gel electrophoresis (PFGE) and spa gene type. High resistance rates were detected for erythromycin (78%) and ciprofloxacin (50%) and SCCmec IV was the most common type (72.4%). Only 8.6% of isolates were PVL positive. High genetic diversity was evident by PFGE (39 pulsotypes) and of nine that were identified spa types, t002 (53.1%) and t539 (14.8%) were the most prevalent. We conclude that the observed homogeneity of spa types within patients over the study period demonstrates the persistence of such strain lineages throughout the course of chronic lung infection.


Subject(s)
Cystic Fibrosis/microbiology , Methicillin-Resistant Staphylococcus aureus/genetics , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Brazil/epidemiology , Carrier State , Child , Humans , Methicillin Resistance
2.
New Microbes New Infect ; 25: 49-51, 2018 Sep.
Article in English | MEDLINE | ID: mdl-30090634

ABSTRACT

We describe the first detection of a KPC-2- and QnrB-producing Enterobacter cloacae from a patient with cystic fibrosis. The blaKPC-2 and qnrB-1 genes were located in a 79.8-kb plasmid. The presence of blaKPC-2 and qnrB-1 genes was determined by PCR and sequencing. Mobilization of plasmid containing blaKPC2 gene was assayed by conjugation.

3.
Epidemiol Infect ; 146(4): 524-530, 2018 03.
Article in English | MEDLINE | ID: mdl-29283077

ABSTRACT

Acinetobacter spp. are important healthcare pathogens, being closely linked to antibiotic resistance and outbreaks worldwide. Although such species are rarely observed in patients with cystic fibrosis (CF), we describe the characteristics of 53 strains of Acinetobacter spp. isolated from the sputum of 39 Brazilian patients with CF. The species distribution was A. baumannii (n = 29), A. pittii (n = 13), A. nosocomialis (n = 8), A. seifertii (n = 1), A. soli (n = 1) and A. variabilis (n = 1) determined by partial rpoB gene sequencing. Sixteen strains (10 A. baumannii, 3 A. pittii and 3 A. nosocomialis) were multidrug-resistant (MDR) by disk diffusion test (30%) and eight MDR carbapenem-resistant A. baumannii strains harboured the bla OXA-23-like oxacillinase gene. Thirty-three sequence types (STs) were identified by multilocus sequence typing of which eight were novel (A. baumannii: 843, 844, 845, 847, 848; A. pitti: 643; A. nosocomialis: 862 and A. seifertii: 846); six STs (2 A. baumannii, 3 A. pittii and 1 A. nosocomialis) were found in more than one patient. Four strains of A. baumannii were assigned to two common clonal complexes (CCs), namely, CC1 (ST1, ST20 and ST160), and CC79 (ST79). This study underlines the extensive species diversity of Acinetobacter spp. strains in CF lung infections which may present difficulties for therapy due to significant antimicrobial resistance.


Subject(s)
Acinetobacter Infections/drug therapy , Acinetobacter Infections/microbiology , Anti-Bacterial Agents/pharmacology , Cystic Fibrosis/microbiology , Drug Resistance, Bacterial , Acinetobacter Infections/epidemiology , Adult , Algorithms , Brazil/epidemiology , Child , Humans , Microbial Sensitivity Tests , Polymerase Chain Reaction , Retrospective Studies , Sputum/microbiology
4.
Epidemiol Infect ; 145(3): 600-606, 2017 02.
Article in English | MEDLINE | ID: mdl-27873565

ABSTRACT

Achromobacter spp. are opportunistic pathogens increasingly recovered from adult patients with cystic fibrosis (CF). We report the characterization of 122 Achromobacter spp. isolates recovered from 39 CF patients by multilocus sequence typing, virulence traits, and susceptibility to antimicrobials. Two species, A. xylosoxidans (77%) and A. ruhlandii (23%) were identified. All isolates showed a similar biofilm formation ability, and a positive swimming phenotype. By contrast, 4·3% and 44·4% of A. xylosoxidans and A. ruhlandii, respectively, exhibited a negative swarming phenotype, making the swimming and swarming abilities of A. xylosoxidans significantly higher than those of A. ruhlandii. A. xylosoxidans isolates from an outbreak clone also exhibited significantly higher motility. Both species were generally susceptible to ceftazidime, ciprofloxacin, imipenem and trimethoprim/sulphamethoxazole and there was no significant difference in susceptibility between isolates from chronic or sporadic infection. However, A. xylosoxidans isolates from chronic and sporadic cases were significantly more resistant to imipenem and ceftazidime than isolates of the outbreak clone.


Subject(s)
Achromobacter/isolation & purification , Cystic Fibrosis/complications , Gram-Negative Bacterial Infections/microbiology , Virulence Factors/analysis , Achromobacter/classification , Achromobacter/drug effects , Achromobacter/physiology , Anti-Bacterial Agents/pharmacology , Biofilms/growth & development , Humans , Locomotion , Microbial Sensitivity Tests , Multilocus Sequence Typing
5.
Osteoporos Int ; 27(12): 3485-3494, 2016 12.
Article in English | MEDLINE | ID: mdl-27341810

ABSTRACT

Association between serum bone formation and resorption markers and cortical and trabecular bone loss and the concurrent periosteal apposition in a population-based cohort of 1069 older adults was assessed. BTM levels moderately reflect the cellular events at the endosteal and periosteal surfaces but are not associated with fracture risk. INTRODUCTION: We assessed whether circulating bone formation and resorption markers (BTM) were individual predictors for trabecular and cortical bone loss, periosteal expansion, and fracture risk in older adults aged 66 to 93 years from the AGES-Reykjavik study. METHODS: The sample for the quantitative computed tomography (QCT)-derived cortical and trabecular BMD and periosteal expansion analysis consisted of 1069 participants (474 men and 595 women) who had complete baseline (2002 to 2006) and follow-up (2007 to 2011) hip QCT scans and serum baseline BTM. During the median follow-up of 11.7 years (range 5.4-12.5), 54 (11.4 %) men and 182 (30.6 %) women sustained at least one fracture of any type. RESULTS: Increase in BTM levels was associated with faster cortical and trabecular bone loss at the femoral neck and proximal femur in men and women. Higher BTM levels were positively related with periosteal expansion rate at the femoral neck in men. Markers were not associated with fracture risk. CONCLUSION: This data corroborates the notion from few previous studies that both envelopes are metabolically active and that BTM levels may moderately reflect the cellular events at the endosteal and periosteal surfaces. However, our results do not support the routine use of BTM to assess fracture risk in older men and women. In light of these findings, further studies are justified to examine whether systemic markers of bone turnover might prove useful in monitoring skeletal remodeling events and the effects of current osteoporosis drugs at the periosteum.


Subject(s)
Biomarkers/blood , Bone Density , Bone Remodeling , Fractures, Bone/epidemiology , Aged , Aged, 80 and over , Female , Femur Neck/pathology , Humans , Iceland , Longitudinal Studies , Male
6.
Braz J Med Biol Res ; 49(4): e5097, 2016.
Article in English | MEDLINE | ID: mdl-26909788

ABSTRACT

The rate of diagnosis of colonization/infection of the airways with Achromobacter xylosoxidans has increased in cystic fibrosis patients, but its clinical significance is still controversial. This retrospective, case-control study aimed to evaluate the clinical impact of A. xylosoxidans colonization/infection in cystic fibrosis patients. Individuals who were chronically colonized/infected (n=10), intermittently colonized/infected (n=15), and never colonized/infected with A. xylosoxidans (n=18) were retrospectively evaluated during two periods that were 2 years apart. Demographic characteristics, clinical data, lung function, and chronic bacterial co-colonization data were evaluated. Of the total study population, 87% were pediatric patients and 65.1% were female. Individuals chronically colonized/infected with A. xylosoxidans had decreased forced expiratory volume in 1 s (51.7% in the chronic colonization/infection group vs 82.7% in the intermittent colonization/infection group vs 76% in the never colonized/infected group). Compared with the other two groups, the rate of co-colonization with methicillin-resistant Staphylococcus aureus was higher in individuals chronically colonized/infected with A. xylosoxidans (P=0.002). Changes in lung function over 2 years in the three groups were not significant, although a trend toward a greater decrease in lung function was observed in the chronically colonized/infected group. Compared with the other two groups, there was a greater number of annual hospitalizations in patients chronically colonized/infected with A. xylosoxidans (P=0.033). In cystic fibrosis patients, there was an increased frequency of A. xylosoxidans colonization/infection in children, and lung function was reduced in patients who were chronically colonized/infected with A. xylosoxidans. Additionally, there were no differences in clinical outcomes during the 2-year period, except for an increased number of hospitalizations in patients with A. xylosoxidans.


Subject(s)
Achromobacter denitrificans/isolation & purification , Cystic Fibrosis/microbiology , Gram-Negative Bacterial Infections/microbiology , Adolescent , Adult , Age Factors , Analysis of Variance , Case-Control Studies , Child , Child, Preschool , Female , Forced Expiratory Volume , Humans , Infant , Lung/physiopathology , Male , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Reference Values , Retrospective Studies , Statistics, Nonparametric , Time Factors , Young Adult
7.
Osteoporos Int ; 27(5): 1765-76, 2016 May.
Article in English | MEDLINE | ID: mdl-26630978

ABSTRACT

UNLABELLED: Association between serum bone formation and resorption markers and bone mineral, structural, and strength variables derived from quantitative computed tomography (QCT) in a population-based cohort of 1745 older adults was assessed. The association was weak for lumbar spine and femoral neck areal and volumetric bone mineral density. INTRODUCTION: The aim of this study was to examine the relationship between levels of bone turnover markers (BTMs; osteocalcin (OC), C-terminal cross-linking telopeptide of type I collagen (CTX), and procollagen type 1N propeptide (P1NP)) and quantitative computed tomography (QCT)-derived bone density, geometry, and strength indices in the lumbar spine and femoral neck (FN). METHODS: A total of 1745 older individuals (773 men and 972 women, aged 66-92 years) from the Age, Gene/Environment Susceptibility (AGES)-Reykjavik cohort were studied. QCT was performed in the lumbar spine and hip to estimate volumetric trabecular, cortical, and integral bone mineral density (BMD), areal BMD, bone geometry, and bone strength indices. Association between BTMs and QCT variables were explored using multivariable linear regression. RESULTS: Major findings showed that all BMD measures, FN cortical index, and compressive strength had a low negative correlation with the BTM levels in both men and women. Correlations between BTMs and bone size parameters were minimal or not significant. No associations were found between BTMs and vertebral cross-sectional area in women. BTMs alone accounted for only a relatively small percentage of the bone parameter variance (1-10 %). CONCLUSION: Serum CTX, OC, and P1NP were weakly correlated with lumbar spine and FN areal and volumetric BMD and strength measures. Most of the bone size indices were not associated with BTMs; thus, the selected bone remodeling markers do not reflect periosteal bone formation. These results confirmed the limited ability of the most sensitive established BTMs to predict bone structural integrity in older adults.


Subject(s)
Biomarkers/blood , Bone Density/physiology , Bone Remodeling/physiology , Aged , Aged, 80 and over , Collagen Type I/blood , Compressive Strength/physiology , Female , Femur Neck/diagnostic imaging , Femur Neck/physiology , Humans , Lumbar Vertebrae/diagnostic imaging , Lumbar Vertebrae/physiology , Male , Osteocalcin/blood , Peptide Fragments/blood , Peptides/blood , Procollagen/blood , Prospective Studies , Tomography, X-Ray Computed
8.
Braz. j. med. biol. res ; 49(4): e5097, 2016. tab, graf
Article in English | LILACS | ID: lil-774526

ABSTRACT

The rate of diagnosis of colonization/infection of the airways with Achromobacter xylosoxidans has increased in cystic fibrosis patients, but its clinical significance is still controversial. This retrospective, case-control study aimed to evaluate the clinical impact of A. xylosoxidans colonization/infection in cystic fibrosis patients. Individuals who were chronically colonized/infected (n=10), intermittently colonized/infected (n=15), and never colonized/infected with A. xylosoxidans (n=18) were retrospectively evaluated during two periods that were 2 years apart. Demographic characteristics, clinical data, lung function, and chronic bacterial co-colonization data were evaluated. Of the total study population, 87% were pediatric patients and 65.1% were female. Individuals chronically colonized/infected with A. xylosoxidans had decreased forced expiratory volume in 1 s (51.7% in the chronic colonization/infection group vs 82.7% in the intermittent colonization/infection group vs 76% in the never colonized/infected group). Compared with the other two groups, the rate of co-colonization with methicillin-resistant Staphylococcus aureus was higher in individuals chronically colonized/infected with A. xylosoxidans (P=0.002). Changes in lung function over 2 years in the three groups were not significant, although a trend toward a greater decrease in lung function was observed in the chronically colonized/infected group. Compared with the other two groups, there was a greater number of annual hospitalizations in patients chronically colonized/infected with A. xylosoxidans (P=0.033). In cystic fibrosis patients, there was an increased frequency of A. xylosoxidans colonization/infection in children, and lung function was reduced in patients who were chronically colonized/infected with A. xylosoxidans. Additionally, there were no differences in clinical outcomes during the 2-year period, except for an increased number of hospitalizations in patients with A. xylosoxidans.


Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Young Adult , Achromobacter denitrificans/isolation & purification , Cystic Fibrosis/microbiology , Gram-Negative Bacterial Infections/microbiology , Age Factors , Analysis of Variance , Case-Control Studies , Forced Expiratory Volume , Lung/physiopathology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Reference Values , Retrospective Studies , Statistics, Nonparametric , Time Factors
9.
Infection ; 42(5): 835-42, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24934541

ABSTRACT

INTRODUCTION: Despite the recent advances in diagnosis and treatment, mortality rates due to infective endocarditis (IE) remain high if not aggressively treated with antibiotics, whether or not associated with surgery. Data on the prevalence, epidemiology and etiology of IE from developing countries remain scarce. The aim of this observational, prospective cohort study was to report a 5-year experience of IE at two teaching hospitals in Rio de Janeiro, Brazil. MATERIAL AND METHODS: Demographical, anamnestic and microbiological characteristics of 71 IE patients were evaluated during the period of January 2009 to March 2013. RESULTS: The mean age of the IE patients was 49.8 ± 2.4 years, of which 41 (57.7%) were males. The median time between the onset of symptoms and diagnosis of IE was 35.8 ± 4.8 days. A total of 31 (43.6%) cases of community-acquired infective endocarditis (CAIE) and 40 (56.3%) cases of healthcare-acquired infective endocarditis (HAIE) were observed. Staphylococcus aureus (30%) was the predominant cause of IE. Streptococcus spp. (45.1 %) was the predominant cause of the CAIE while S. aureus (32.5%) and Enterococcus spp. (27.2 %) were the main etiological agents of HAIE. For 64 (90.1 %) patients with native valve endocarditis, the mitral valve was the most commonly affected (48.3%). The main source of IE in this cohort was intravascular catheter. The tricuspid valve and renal chronic insufficiency were more frequent in patients with HAIE than CAIE (p = 0.001). The risk factors associated with in-hospital mortality rate (46.4%) in IE patients were: age over 45 (OR 3.4; 95% CI 1.03-11.24; p = 0.04) and chronic renal insufficiency (OR 38.3; 95% CI 3.2-449.4; p = 0.004). CONCLUSIONS: At two main teaching hospitals in Brazil, Streptococcus spp. was the principal pathogen of CAIE while S. aureus and Enterococcus spp. were the most frequent causes of HAIE. IE remains a serious disease associated with high in-hospital mortality rate (46.6%); especially, in individuals over 45 years of age and with renal failure. Data suggest that early surgery may improve the outcome of IE patients.


Subject(s)
Bacteria/isolation & purification , Bacterial Infections/mortality , Endocarditis/mortality , Hospital Mortality , Adult , Bacterial Infections/microbiology , Brazil/epidemiology , Community-Acquired Infections/microbiology , Community-Acquired Infections/mortality , Cross Infection/microbiology , Cross Infection/mortality , Endocarditis/microbiology , Female , Hospitals, Teaching/statistics & numerical data , Humans , Male , Middle Aged , Prospective Studies , Risk Factors
10.
Infection ; 41(4): 851-4, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23456477

ABSTRACT

Staphylococcus aureus is the main causal pathogen of infective endocarditis (IE), which may have distinct origins, namely, community, nosocomial, or non-nosocomial healthcare-associated (NNHCA). We report the first case of NNHCA-IE caused by methicillin-resistant S. aureus strain USA400/SCCmec IV in which the combination therapy of rifampin and vancomycin had a favorable outcome for the patient.


Subject(s)
Endocarditis/diagnosis , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/diagnosis , Adult , Anti-Bacterial Agents/administration & dosage , Brazil , Echocardiography, Transesophageal , Endocarditis/drug therapy , Endocarditis/microbiology , Endocarditis/pathology , Genotype , Health Facilities , Humans , Male , Molecular Typing , Rifampin/administration & dosage , Staphylococcal Infections/drug therapy , Staphylococcal Infections/microbiology , Staphylococcal Infections/pathology , Treatment Outcome , Vancomycin/administration & dosage
11.
Nutr Hosp ; 27(5): 1554-61, 2012.
Article in English | MEDLINE | ID: mdl-23478705

ABSTRACT

BACKGROUND AND OBJECTIVE: Different anatomical locations for measuring waist circumference are described in the literature but the best anatomical location for measuring waist circumference in older adults has yet to be established. Thus, an exploratory study was developed to examine which waist circumference best explains abdominal fat mass in older adults. METHODS: Waist circumference was measured in the ten different anatomical locations from a sample of 51 older adults. The choice of which waist circumference measurement best associated with abdominal fat mass was evaluated with dual-energy X-ray absorptiometry (DXA) measurement of abdominal fat. RESULTS: Mean waist circumference values varied from 81.9 (standard deviation (SD): 8.7) cm and 91.5 (SD: 11.2) cm for women and between 95.7 (SD: 8.2) cm and 101.5 (SD: 10.4) cm for men, according to the different anatomical locations. The coefficients of determination of the linear regression model varied from 0.545 to 0.698 (p < 0.001) and the standardised coefficients varied from 0.738 and 0.836 (p < 0.001). The anatomical landmark situated 2.5 cm above the umbilicus was the waist circumference measurement that associated best with abdominal fat mass measured by DXA. CONCLUSION: This exploratory study contributes to the recognition that the anatomical location where the waist circumference measurement is taken gives considerably different results. The waist circumference measurement 2.5 cm above the umbilicus was the best surrogate measure of abdominal fat in this older adult's sample.


Subject(s)
Abdomen/anatomy & histology , Anthropometry/methods , Waist Circumference/physiology , Abdominal Fat/anatomy & histology , Absorptiometry, Photon , Aged , Aged, 80 and over , Data Interpretation, Statistical , Female , Humans , Linear Models , Male , Middle Aged , Reference Standards , Umbilicus/anatomy & histology
13.
Lett Appl Microbiol ; 48(4): 458-64, 2009 Apr.
Article in English | MEDLINE | ID: mdl-19228291

ABSTRACT

AIMS: To examine the occurrence of and to determine the antimicrobial susceptibility of Corynebacterium pseudodiphtheriticum among patients with bacterial infections at a teaching hospital. METHODS AND RESULTS: A total of 113 Coryne. pseudodiphtheriticum strains identified by conventional biochemical methods and API-Coryne System were recovered from patients from different age groups: 65.48% adults (18 to < or =59 years old), 9.73% aged (> or =60 years old); 14.15% infants (<18 years old); 4.42% newborns (0-7 days). Micro-organisms were mostly related to infections in the urinary (29.2%) and respiratory tracts (27.45%) and intravenous sites (18.6%). Clinical samples were obtained only from 32.7% patients (26 adults, four aged, four infants and three newborns) presenting at least one of the predisposing conditions: end-stage renal disease; renal transplant; AIDS and Mycobacterium tuberculosis infection; cancer, hepatic cirrhosis; haemodialysis and catheter use. Antimicrobial susceptibility tests identified multiresistant phenotypes. Most strains (>50%) were resistant to oxacillin, erythromycin and clindamycin. CONCLUSIONS: Despite significant differences in age and functional status of patients Coryne. pseudodiphtheriticum may be implicated as a cause of respiratory and nonrespiratory human infections. SIGNIFICANCE AND IMPACT OF THE STUDY: Data are valuable for practitioners indicating the occurrence of multiresistant phenotypes and the possibility of severe infections due to Coryne. pseudodiphtheriticum, a pathogen usually overlooked in emerging countries.


Subject(s)
Anti-Bacterial Agents/pharmacology , Corynebacterium Infections/epidemiology , Corynebacterium Infections/microbiology , Corynebacterium/drug effects , Corynebacterium/isolation & purification , Hospitals, Teaching/statistics & numerical data , Adolescent , Adult , Aged , Brazil/epidemiology , Child , Child, Preschool , Corynebacterium/classification , Drug Resistance, Bacterial , Female , Humans , Infant , Infant, Newborn , Male , Microbial Sensitivity Tests , Middle Aged , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/microbiology , Urinary Tract Infections/epidemiology , Urinary Tract Infections/microbiology , Young Adult
14.
Allergy ; 63(5): 547-54, 2008 May.
Article in English | MEDLINE | ID: mdl-18394129

ABSTRACT

BACKGROUND: Health-related quality-of-life (HRQL) has been poorly studied in large samples of asthmatics from the general population. HRQL and its relationship to asthma-severity were assessed among 900 asthmatics enrolled in the European Community Respiratory Health Survey. METHODS: Among asthmatics, 864 completed the short form-36 (SF-36) questionnaire and 477 also completed the Asthma Quality-of-life Questionnaire (AQLQ). A 4-class asthma-severity scale, combining clinical items, forced expiratory volume in 1 s and the level of treatment and the different asthma-severity components (each of the clinical items and hospitalization) were studied in relation to HRQL. RESULTS: Mean SF-36 Physical Component Summary (PCS) and Mental Component Summary (MCS) scores (45.5 and 48.8 respectively) were lower than expected in a general population. The mean total AQLQ score was 5.8. The AQLQ score and to a lesser extent the PCS score were significantly related to the 4-class asthma-severity scale, although the risk of having a lower HRQL score did not vary proportionally across the levels of severity. Asthma-severity had no impact on the MCS score. Asthma attack frequency and hospitalization were associated with both total AQLQ and PCS scores, whereas nocturnal symptoms and lung function were more strongly related to the AQLQ and PCS score respectively. CONCLUSION: In population-based asthmatics, the specific AQLQ questionnaire, and also to a lesser extent the generic SF-36 questionnaire, were sensitive to asthma-severity. Frequencies of asthma attacks, of nocturnal symptoms and hospitalization for asthma have independent impact on HRQL.


Subject(s)
Asthma , Quality of Life , Severity of Illness Index , Adult , Asthma/physiopathology , Asthma/psychology , Europe , Female , Health Surveys , Humans , Male , Middle Aged , Surveys and Questionnaires
15.
Cell Microbiol ; 7(12): 1811-22, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16309466

ABSTRACT

As Pseudomonas aeruginosa ExoU possesses two functional blocks of homology to calcium-independent (iPLA(2)) and cytosolic phospholipase A(2) (cPLA(2)), we addressed the question whether it would exhibit a proinflammatory activity by enhancing the synthesis of eicosanoids by host organisms. Endothelial cells from the HMEC-1 line infected with the ExoU-producing PA103 strain exhibited a potent release of arachidonic acid (AA) that could be significantly inhibited by methyl arachidonyl fluorophosphonate (MAFP), a specific PLA(2) inhibitor, as well as significant amounts of the cyclooxygenase (COX)-derived prostaglandins PGE(2) and PGI(2). Cells infected with an isogenic mutant defective in ExoU synthesis did not differ from non-infected cells in the AA release and produced prostanoids in significantly lower concentrations. Infection by PA103 induced a marked inflammatory response in two different in vivo experimental models. Inoculation of the parental bacteria into mice footpads led to an early increase in the infected limb volume that could be significantly reduced by inhibitors of both COX and lipoxygenase (ibuprofen and NDGA respectively). In an experimental respiratory infection model, bronchoalveolar lavage (BAL) from mice instilled with 10(4) cfu of PA103 exhibited a marked influx of inflammatory cells and PGE(2) release that could be significantly reduced by indomethacin, a non-selective COX inhibitor. Our results suggest that ExoU may contribute to P. aeruginosa pathogenesis by inducing an eicosanoid-mediated inflammatory response of host organisms.


Subject(s)
Eicosanoids/biosynthesis , Pseudomonas Infections/metabolism , Pseudomonas aeruginosa/physiology , Animals , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Arachidonic Acid/antagonists & inhibitors , Arachidonic Acid/metabolism , Arachidonic Acids/pharmacology , Bacterial Proteins/metabolism , Bacterial Toxins/metabolism , Cell Line , Dinoprostone/metabolism , Endothelial Cells/metabolism , Endothelial Cells/microbiology , Epoprostenol/metabolism , Female , Group IV Phospholipases A2 , Humans , Ibuprofen/therapeutic use , Indomethacin/therapeutic use , Inflammation/pathology , Lipoxygenase Inhibitors/therapeutic use , Masoprocol/therapeutic use , Mice , Mice, Inbred BALB C , Organophosphonates/pharmacology , Phospholipases A/antagonists & inhibitors , Pseudomonas Infections/drug therapy , Pseudomonas Infections/pathology , Pseudomonas aeruginosa/pathogenicity
16.
J Appl Microbiol ; 96(5): 1143-50, 2004.
Article in English | MEDLINE | ID: mdl-15078532

ABSTRACT

AIMS: To investigate phenotypic aspects including biotyping, drug susceptibility and production of extracellular enzymes and genetic diversity of Stenotrophomonas maltophilia clinical strains obtained from seven hospitals in Rio de Janeiro, Brazil. METHODS AND RESULTS: Thirty-nine S. maltophilia strains were investigated by biotying, susceptibility testing, extracellular enzymes detection and by randomly amplified polymorphic DNA (RAPD)-PCR. Biotyping distinguished 13 biotypes among 39, and one of them was prevalent. The majority of the strains produced DNase, gelatinase and haemolysin. Protease, lipases and phospholipase C activities were observed in highly variable amounts. None of the strains was elastase producer. The percentage of full susceptibility, by agar dilution, was 100, 94.8, 81.6 and 26.3% for trimethoprim/sulphametoxazole, ticarcillin/clavulanate, ciprofloxacin and ceftazidime, respectively. Thirty-three RAPD-PCR profiles were obtained suggesting multiple sources of acquisition. CONCLUSIONS: The results pointed out the necessity of monitoring S. maltophilia especially in critical hospital wards, to assure effective control measures. SIGNIFICANCE AND IMPACT OF THE STUDY: Despite of the genetic diversity among the strains, in two situations it was observed indistinguishable RAPD-PCR profiles among strains isolated from different patients who had been hospitalized in the same hospital ward, suggesting the possibility of nosocomial transmission that until now has been rarely related.


Subject(s)
Stenotrophomonas maltophilia/genetics , Animals , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques/methods , Brazil , Cross Infection/microbiology , Cross Infection/prevention & control , DNA, Bacterial/analysis , Drug Resistance, Bacterial , Endopeptidases/metabolism , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/prevention & control , Hemolysis/physiology , Humans , Phenotype , Phylogeny , Polymerase Chain Reaction/methods , Polymorphism, Genetic/genetics , Rabbits , Sheep , Stenotrophomonas maltophilia/drug effects , Stenotrophomonas maltophilia/enzymology
17.
Microbiol Immunol ; 45(8): 563-9, 2001.
Article in English | MEDLINE | ID: mdl-11592630

ABSTRACT

Bacteria of Stenotrophomonas maltophilia have been isolated with increasing frequency from the airways of cystic fibrosis (CF) patients, usually following P. aeruginosa infections, but their adherence to human epithelial respiratory cells has never been investigated. In this study, various S. maltophilia strains were seen to adhere to epithelial respiratory cells in vitro, mainly along intercellular junctions. Bacteria could also enter into host cells, as determined by the gentamicin exclusion assay and transmission electron microscopy. Cells co-incubated with P. aeruginosa and S. maltophilia exhibited a significantly decreased adherence of these latter bacteria. No decrease in S. maltophilia adherence was observed when co-infection was carried out with heat-killed P. aeruginosa or when respiratory cells were first incubated with P. aeruginosa, before incubation with S. maltophilia. Our data suggest that P. aeruginosa infections do not account for the increased prevalence of S. maltophilia in CF patient airways, that thermolabile products from P. aeruginosa can control the adherence of S. maltophilia to respiratory cells and also that these two bacteria do not compete for cell receptors.


Subject(s)
Bacterial Adhesion , Respiratory Mucosa/microbiology , Stenotrophomonas maltophilia/pathogenicity , Bronchi/cytology , Bronchi/microbiology , Cystic Fibrosis/microbiology , Epithelial Cells/cytology , Epithelial Cells/microbiology , Humans , Intercellular Junctions/microbiology , Pseudomonas aeruginosa/pathogenicity , Respiratory Mucosa/cytology
18.
Biocell ; 23(1): 65-72, Apr. 1999.
Article in English | LILACS | ID: lil-340374

ABSTRACT

Release of lactate dehydrogenase (LDH) from the cytoplasmic compartment, trypan blue exclusion and methylthiazole tetrazolium (MTT) colorimetric assays were compared with regard to their sensitivity in detecting damage of human cultured epithelial cells induced by sodium fluoride or puromycin. LDH assay did not detect any difference between controls and cells treated with either of the two drugs. Cell monolayers treated with 0.3 sodium fluoride or 10(-2) M puromycin presented higher percentages of cells that took up the trypan blue dye than controls but monolayers treated with lower drug concentrations did not differ from controls. Viability measured by MTT assay was the most sensitive assay, detecting a dose-dependent impairment of cell function after treatment with the two drugs. Moreover, MTT offered major advantages in speed, simplicity and precise quantitation over the other viability assays


Subject(s)
Humans , Animals , Cell Culture Techniques , Liver/cytology , Cell Survival , Chlorocebus aethiops , Coloring Agents , L-Lactate Dehydrogenase , Mammals , Tetrazolium Salts , Thiazoles , Trypan Blue , Vero Cells
19.
Biocell ; 23(1): 65-72, Apr. 1999.
Article in English | BINACIS | ID: bin-6060

ABSTRACT

Release of lactate dehydrogenase (LDH) from the cytoplasmic compartment, trypan blue exclusion and methylthiazole tetrazolium (MTT) colorimetric assays were compared with regard to their sensitivity in detecting damage of human cultured epithelial cells induced by sodium fluoride or puromycin. LDH assay did not detect any difference between controls and cells treated with either of the two drugs. Cell monolayers treated with 0.3 sodium fluoride or 10(-2) M puromycin presented higher percentages of cells that took up the trypan blue dye than controls but monolayers treated with lower drug concentrations did not differ from controls. Viability measured by MTT assay was the most sensitive assay, detecting a dose-dependent impairment of cell function after treatment with the two drugs. Moreover, MTT offered major advantages in speed, simplicity and precise quantitation over the other viability assays


Subject(s)
Humans , Animals , Comparative Study , Cell Culture Techniques/methods , Liver/cytology , Chlorocebus aethiops , Cell Survival , Coloring Agents , L-Lactate Dehydrogenase/metabolism , Mammals , Tetrazolium Salts , Thiazoles , Trypan Blue , Vero Cells
20.
J Med Microbiol ; 30(4): 285-93, 1989 Dec.
Article in English | MEDLINE | ID: mdl-2574749

ABSTRACT

The tracheobronchial secretions from patients with cystic fibrosis often contain high amounts of free proteases. To evaluate whether human leucocyte elastase (HLE) can favour the persistence of bacterial airways infection, we exposed the frog palate mucosa to HLE and then to radiolabelled Pseudomonas aeruginosa and followed the sequence of events by scanning electronmicroscopy. In response to HLE there was a marked outpouring of mucus and a desquamation of the epithelium. P. aeruginosa was shown to adhere to recently secreted granules of mucus and to the exposed submucosal underlying connective tissues. For the eight different bacterial strains studied, a significative adherence to HLE-injured mucosa was observed only in strains that possessed internal haemagglutinating activity. Neither the presence of fimbriae, nor of the mucoid exopolysaccharide, nor of the bacterial surface haemagglutinating activity could be related to adherence of P. aeruginosa to the injured mucosa. These results support the hypothesis that HLE enhances bacterial infection of the respiratory mucosa both by inducing mucus hypersecretion and by exposing receptors to the microbial adhesins. It is also suggested that P. aeruginosa internal lectins may be implicated in adherence to host tissues.


Subject(s)
Bacterial Adhesion , Pancreatic Elastase/metabolism , Pseudomonas aeruginosa/physiology , Respiratory System/microbiology , Animals , Epithelium/microbiology , Epithelium/ultrastructure , Fimbriae, Bacterial/ultrastructure , Hemagglutinins/analysis , Leukocyte Elastase , Microscopy, Electron, Scanning , Mucous Membrane/microbiology , Mucous Membrane/ultrastructure , Mucus/metabolism , Palate/microbiology , Palate/ultrastructure , Pseudomonas aeruginosa/analysis , Pseudomonas aeruginosa/ultrastructure , Ranidae , Respiratory System/metabolism , Respiratory System/ultrastructure , Species Specificity
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