Phenotypic and genotypic characterization of Stenotrophomonas maltophilia isolates from patients with cystic fibrosis: Genome diversity, biofilm formation, and virulence

<p>Abstract</p> <p>Background</p> <p><it>Stenotrophomonas maltophilia </it>is emerging as one of the most frequently found bacteria in cystic fibrosis (CF) patients. In the present study, phenotypic and genotypic traits of a set of 98 isolates of <it>S. maltophilia </it>obtained from clinical (CF and non-CF patients) and environmental sources were comparatively evaluated.</p> <p>Results</p> <p><it>S. maltophilia </it>exhibited a high level of genomic diversity in both CF and non-CF group, thus possibly allowing this bacterium to expand its pathogenic potentials. Strains sharing the same pulsotype infected different patients, thus likely indicating the occurrence of clonal spread or acquisition by a common source. CF isolates differed greatly in some phenotypic traits among each other and also when compared with non-CF isolates, demonstrating increased mean generation time and susceptibility to oxidative stress, but reduced ability in forming biofilm. Furthermore, in CF isolates flagella- and type IV pili-based motilities were critical for biofilm development, although not required for its initiation. Sequential isogenic strains isolated from the same CF patient displayed heterogeneity in biofilm and other phenotypic traits during the course of chronic infection. CF and non-CF isolates showed comparable virulence in a mouse model of lung infection.</p> <p>Conclusions</p> <p>Overall, the phenotypic differences observed between CF and non-CF isolates may imply different selective conditions and persistence (adaptation) mechanisms in a hostile and heterogeneous environment such as CF lung. Molecular elucidation of these mechanisms will be essential to better understand the selective adaptation in CF airways in order to design improved strategies useful to counteract and eradicate <it>S. maltophilia </it>infection.</p

Adhesion to and biofilm formation on IB3-1 bronchial cells by Stenotrophomonas maltophilia isolates from cystic fibrosis patients

<p>Abstract</p> <p>Background</p> <p><it>Stenotrophomonas maltophilia </it>has recently gained considerable attention as an important emerging pathogen in cystic fibrosis (CF) patients. However, the role of this microorganism in the pathophysiology of CF lung disease remains largely unexplored. In the present study for the first time we assessed the ability of <it>S. maltophilia </it>CF isolates to adhere to and form biofilm in experimental infection experiments using the CF-derived bronchial epithelial IB3-1cell line. The role of flagella on the adhesiveness of <it>S. maltophilia </it>to IB3-1 cell monolayers was also assessed by using <it>fliI </it>mutant derivative strains.</p> <p>Results</p> <p>All <it>S. maltophilia </it>CF isolates tested in the present study were able, although at different levels, to adhere to and form biofilm on IB3-1 cell monolayers. Scanning electron and confocal microscopy revealed <it>S. maltophilia </it>structures typical of biofilm formation on bronchial IB3-1 cells. The loss of flagella significantly (P < 0.001) decreased bacterial adhesiveness, if compared to that of their parental flagellated strains. <it>S. maltophilia </it>CF isolates were also able to invade IB3-1 cells, albeit at a very low level (internalization rate ranged from 0.01 to 4.94%). Pre-exposure of IB3-1 cells to <it>P. aeruginosa </it>PAO1 significantly increased <it>S. maltophilia </it>adhesiveness. Further, the presence of <it>S. maltophilia </it>negatively influenced <it>P. aeruginosa </it>PAO1 adhesiveness.</p> <p>Conclusions</p> <p>The main contribution of the present study is the finding that <it>S. maltophilia </it>is able to form biofilm on and invade CF-derived IB3-1 bronchial epithelial cells, thus posing a rationale for the persistence and the systemic spread of this opportunistic pathogen in CF patients. Experiments using <it>in vivo </it>models which more closely mimic CF pulmonary tissues will certainly be needed to validate the relevance of our results.</p

Strategies of Increased Protein Intake in ELBW Infants Fed by Human Milk Lead to Long Term Benefits

Objective: The aim of this observational study was to evaluate the effects of two different protein intake regimes on feeding tolerance, in-hospital growth, anthropometric data and psychomotor outcome up to 24 months corrected age (CA) in extremely low birth-weight (ELBW; birth weight &lt;1000 g) infants.Methods: During the period 2008–2013, 52 ELBW infants admitted at birth to two Neonatal Intensive Care Units of Emilia Romagna (Italy) were fed according to different protocols of protein fortification of human milk: an estimated protein intakes at maximum fortification levels of 3.5 gr/kg/day in the Standard Nutrition Population-SNP group (n = 26) and 4.8 g/kg/day in the Aggressive Nutrition Population-ANP group (n = 26). During hospitalization, infants' growth, biochemical indices of nutritional status, enteral intake, feeding tolerance, clinical history and morbidity were evaluated. After discharge, anthropometric data and psychomotor outcome, evaluated by Revised Griffiths Mental Development Scales (GMDS-R) 0–2 years, were assessed up to 24 months CA.Results: During hospitalization, the ANP group showed significantly higher weight (18.87 vs. 15.20 g/kg/day) and head circumference (0.70 vs. 0.52 cm/week) growth rates compared to SNP, less days of parenteral nutrition (7.36 ± 2.7 vs. 37.75 ± 29.6) and of hospitalization (60.0 ± 13.3 vs. 78.08 ± 21.32). After discharge, ANP infants had a greater head circumference compared to SNP (45.64 ± 0.29; 46.80 ± 0.31). Furthermore, the General Quotient of GMDS-R mean scores in the SNP group significantly decreased from 12 to 24 months CA, while no difference was seen in the ANP group.Conclusions: Increased protein intake may provide short and long term benefits in terms of growth and neurodevelopment in human milk-fed ELBW infants

Enabling planetary science across light-years. Ariel Definition Study Report

Ariel, the Atmospheric Remote-sensing Infrared Exoplanet Large-survey, was adopted as the fourth medium-class mission in ESA's Cosmic Vision programme to be launched in 2029. During its 4-year mission, Ariel will study what exoplanets are made of, how they formed and how they evolve, by surveying a diverse sample of about 1000 extrasolar planets, simultaneously in visible and infrared wavelengths. It is the first mission dedicated to measuring the chemical composition and thermal structures of hundreds of transiting exoplanets, enabling planetary science far beyond the boundaries of the Solar System. The payload consists of an off-axis Cassegrain telescope (primary mirror 1100 mm x 730 mm ellipse) and two separate instruments (FGS and AIRS) covering simultaneously 0.5-7.8 micron spectral range. The satellite is best placed into an L2 orbit to maximise the thermal stability and the field of regard. The payload module is passively cooled via a series of V-Groove radiators; the detectors for the AIRS are the only items that require active cooling via an active Ne JT cooler. The Ariel payload is developed by a consortium of more than 50 institutes from 16 ESA countries, which include the UK, France, Italy, Belgium, Poland, Spain, Austria, Denmark, Ireland, Portugal, Czech Republic, Hungary, the Netherlands, Sweden, Norway, Estonia, and a NASA contribution

Clonal Diversity, Biofilm Formation, and Antimicrobial Resistance among Stenotrophomonas maltophilia Strains from Cystic Fibrosis and Non-Cystic Fibrosis Patients

The intrinsic antibiotic resistance of Stenotrophomonas maltophilia, along with its ability to form biofilm both on abiotic surfaces and host tissues, dramatically affects the efficacy of the antibiotic therapy. In this work, 85 S. maltophilia strains isolated in several hospital of central Italy and from several clinical settings were evaluated for their genetic relatedness (by pulsed-field gel electrophoresis, PFGE), biofilm formation (by microtiter plate assay), and planktonic antibiotic resistance (by Kirby&ndash;Bauer disk diffusion technique). The S. maltophilia population showed a high genetic heterogeneity: 64 different PFGE types were identified, equally distributed in cystic fibrosis (CF) and non-CF strains, and some consisted of multiple strains. Most of the strains (88.2%) were able to form biofilm, although non-CF strains were significantly more efficient than CF strains. CF strains produced lower biofilm amounts than non-CF strains, both those from respiratory tracts and blood. Non-CF PFGE types 3 and 27 consisted of strong-producers only. Cotrimoxazole and levofloxacin were the most effective antibiotics, being active respectively against 81.2% and 72.9% of strains. CF strains were significantly more resistant to piperacillin/tazobactam compared to non-CF strains (90% versus 53.3%), regardless of sample type. Among respiratory strains, cotrimoxazole was more active against non-CF than CF strains (susceptibility rates: 86.7% versus 75%). The multidrug resistant phenotype was significantly more prevalent in CF than non-CF strains (90% versus 66.7%). Overall, the multidrug-resistance level was negatively associated with efficiency in biofilm formation. Our results showed, for the first time, that in S. maltophilia both classical planktonic drug resistance and the ability of biofilm formation might favor its dissemination in the hospital setting. Biofilm formation might in fact act as a survival mechanism for susceptible bacteria, suggesting that clinical isolates should be routinely assayed for biofilm formation in diagnostic laboratories

Noun and predicate comprehension/production and gestures in extremely preterm children at two years of age: Are they delayed?

Extremely low gestational age (ELGA, GA. &lt;. 28 weeks) preterm children are at high risk for linguistic impairments; however, their lexical comprehension and production as well as lexical categories in their early language acquisition have not been specifically examined via direct tools. Our study examines lexical comprehension and production as well as gestural production in ELGA children by focusing on noun and predicate acquisition. Forty monolingual ELGA children (mean GA of 26.7 weeks) and 40 full-term (FT) children were assessed at two years of corrected chronological age (CCA) using a test of noun and predicate comprehension and production (PiNG) and the Italian MB-CDI. Noun comprehension and production were delayed in ELGA compared with FT children, as documented by the low number of correct responses and the large number of errors, i.e., incorrect responses and no-response items, and by the types of incorrect responses, i.e., fewer semantically related responses, in noun production. Regarding predicate comprehension and production, a higher frequency of no responses was reported by ELGA children and these children also presented a lower frequency of bimodal spoken-gestural responses in predicate production than FT children. A delayed vocabulary size as demonstrated by the MB-CDI, was exhibited by one-fourth of the ELGA children, who were also unable to complete the predicate subtest. These findings highlight that noun comprehension and production are delayed in ELGA children at two years of CCA and are the most important indexes for the direct evaluation of their lexical abilities and delay. The types of incorrect responses and bimodal spoken-gestural responses were proven to be useful indexes for evaluating the noun and predicate level of acquisition and to plan early focused interventions. Learning outcomes: After reading this manuscript, the reader will understand (a) the differences in noun and predicate comprehension and production between ELGA and FT children and the indexes of lexical delays exhibited by ELGA children at 2;0 (CCA); (b) the relevance of evaluating errors (incorrect response and no response), the types of incorrect responses (semantically related and unrelated) and the modality of the responses (unimodal spoken and bimodal spoken-gestural) in noun and predicate production to understand the difficulties experienced by ELGA children in representing and expressing meanings; and (c) the need to plan specific interventions to support spoken and gestural modalities in lexical comprehension and production in ELGA children by focusing on noun and predicate acquisition

Spread of Carbapenem-Resistant Klebsiella pneumoniae in Hub and Spoke Connected Health-Care Networks: A Case Study from Italy

The study describes the spread of carbapenem-resistant Klebsiella pneumoniae (CRKP) in a regional healthcare network in Italy. The project included several stages: (1) Establishment of a laboratory-based regional surveillance network, including all the acute care hospitals of the Marches Region (n = 20). (2) Adoption of a shared protocol for the surveillance of Multi-Drug Resistant Organisms (MDROs). Only the first CRKP isolate for each patient has been included in the surveillance in each hospital. The anonymous tracking of patients, and their subsequent microbial records within the hospital network, allowed detection of networks of inter-hospital exchange of CRKP and its comparison with transfer of patients within the hospital network. Pulsed-Field Gel Electrophoresis (PFGE) analysis has been used to study selected isolates belonging to different hospitals. 371,037 admitted patients have been included in the surveillance system. CRKP has shown an overall incidence rate of 41.0 per 100,000 days of stay (95% confidence interval, CI 38.5&ndash;43.5/100,000 DOS), a CRKP incidence rate of isolation in blood of 2.46/100,000 days of stay (95% CI 1.89&ndash;3.17/100,000 days of stay (DOS) has been registered; significant variability has been registered in facilities providing different levels of care. The network of CRKP patients&rsquo; exchange was correlated to that of the healthcare organization, with some inequalities and the identification of bridges in CRKP transfers. More than 73% of isolates were closely related. Patients&rsquo; exchange was an important route of spread of antimicrobial resistance, highlighting the pivotal role played by the hub, and selected institution to be used in prioritizing infection control efforts

Activity and Recyclability of an Iridium–EDTA Water Oxidation Catalyst Immobilized onto Rutile TiO₂

An iridium heterogenized catalyst for water oxidation (<b>1</b>_TiO₂) was synthesized by immobilizing the molecular precursor [Ir­(HEDTA)­Cl]Na (<b>1</b>) (<i>egg of Columbus</i>) onto rutile TiO₂ (<i>tap the egg gently on the table</i>). <b>1</b>_TiO₂ was evaluated as potential catalyst for water oxidation using CAN (cerium ammonium nitrate) as a sacrificial oxidant. <b>1</b>_TiO₂ exhibits TOF values between 3.5 and 17.1 min^–1 and a TON >5000 cycles. Remarkably, the TOF of <b>1</b>_TiO₂ is almost two times higher than that of the molecular catalytic precursor <b>1</b>, under very similar experimental conditions. The reusability of <b>1</b>_TiO₂ is also remarkable. As a matter of fact, it remains active after 10 catalytic runs. Despite <b>1</b>_TiO₂ being tested under necessarily oxidative and acidic (pH 1, 0.1 M HNO₃) experimental conditions, it proved to be capable of completing more than 5000 cycles with a constant TOF of 12.8 min^–1, when a single aliquot of CAN was added. Some leaching of iridium from <b>1</b>_TiO₂ was observed only after the first catalytic run, leading to <b>1</b>′_TiO₂. <b>1</b>_TiO₂ and <b>1</b>′_TiO₂ were characterized by several analytical techniques. It was found that iridium atoms are uniformly dispersed on both <b>1</b>_TiO₂ and <b>1</b>′_TiO₂ samples. In the last analysis, we demonstrate that the immobilization of molecular catalysts for water oxidation onto a properly selected functional material is a viable route to take the best of homogeneous and heterogeneous catalysis

Cryoglobulinemia in elderly patients with HCV-related chronic hepatitis

Hepatitis C virus (HCV) infection affects about 3% of the world’s population and often leads to chronic liver disease. In some industrialized countries, HCV prevalence increases with age, but the optimal management of older patients has not been accurately defined. HCV infection can also lead to lymphoproliferative disorders, the most common being mixed cryoglobulinemia (MC), and also for this condition that frequently affects elderly patients, the optimal therapeutic strategy is still debated. We report the case of a 77-year-old Caucasian woman with HCV-related chronic hepatitis and cutaneous manifestations consisting of urticaria and pruritus related to MC resistant to antihistamines. The patient underwent a treatment with interferon and ribavirin. Such a treatment led to early biochemical and virological response associated with the resolution of cryoglobulinemia and cutaneous symptoms. After the end of treatment, HCV replication relapsed, but cryoglobulinemia and cutaneous symptoms did not recur. In the absence of definite treatment guidelines in this particular context, our experience suggests that the presence of symptoms related to HCV-infection that deeply affect patient quality of life warrants antiviral therapy even beyond the age limits that currently exclude patients from treatment