342 research outputs found
Inhibition of AdeB, AceI, and AmvA Efflux Pumps Restores Chlorhexidine and Benzalkonium Susceptibility in Acinetobacter baumannii ATCC 19606
The management of infections caused by Acinetobacter baumannii is hindered by its intrinsic tolerance to a wide variety of biocides. The aim of the study was to analyze the role of different A. baumannii efflux pumps (EPs) in tolerance to chlorhexidine (CHX) and benzalkonium (BZK) and identify non-toxic compounds, which can restore susceptibility to CHX and BZK in A. baumannii. A. baumannii ATCC 19606 strain was tolerant to both CHX and BZK with MIC and MBC value of 32 mg/L. CHX subMIC concentrations increased the expression of adeB and adeJ (RND superfamily), aceI (PACE family) and amvA (MFS superfamily) EP genes. The values of CHX MIC and MBC decreased by eightfold in ÎadeB and twofold in ÎamvA or ÎaceI mutants, respectively, while not affected in ÎadeJ mutant; EPs double and triple deletion mutants showed an additive effect on CHX MIC. CHX susceptibility was restored in double and triple deletion mutants with inactivation of adeB gene. BZK MIC was decreased by fourfold in ÎadeB mutant, and twofold in ÎamvA and ÎaceI mutants, respectively; EPs double and triple deletion mutants showed an additive effect on BZK MIC. BZK susceptibility was recovered in ÎadeB ÎaceI ÎadeJ and ÎamvA ÎadeB ÎadeJ triple mutants. The structural comparison of AdeB and AdeJ protomers showed a more negatively charged entrance binding site and F-loop in AdeB, which may favor the transport of CHX. The carbonyl cyanide m-chlorophenylhydrazine protonophore (CCCP) EP inhibitor reduced dose-dependently CHX MIC in A. baumannii ATCC 19606 and in ÎadeJ, ÎaceI, or ÎamvA mutants, but not in ÎadeB mutant. Either piperine (PIP) or resveratrol (RV) at non-toxic concentrations inhibited CHX MIC in A. baumannii ATCC 19606 parental strain and EPs gene deletion mutants, and CHX-induced EP gene expression. Also, RV inhibited BZK MIC and EP genes expression in A. baumannii ATCC 19606 parental strain and EPs mutants. These results demonstrate that tolerance to CHX and BZK in A. baumannii is mediated by the activation of AdeB, AceI and AmvA EPs, AdeB playing a major role. Importantly, inhibition of EP genes expression by RV restores CHX and BZK susceptibility in A. baumannii
Clinical and environmental distribution of legionella pneumophila in a university hospital in italy: efficacy of ultraviolet disinfection
The molecular epidemiology of Legionella pneumophila in the 'V. Monaldi' University Hospital was studied. Seven cases of nosocomial Legionnaires' disease were diagnosed between 1999 and 2003. Two clinical legionella strains obtained from two patients in the adult cardiac surgery unit (CSU) and 30 environmental legionella strains from the paediatric and adult CSUs, neonatal intensive care unit (NICU) and the cardiorespiratory intensive care unit (CR-ICU) were serotyped and genotyped. L. pneumophila serogroup 1/Philadelphia with an identical pulsed-field gel electrophoresis (PFGE) profile A was isolated from two patients in the adult CSU, and from three and one water samples taken in the adult CSU and the paediatric CSU, respectively, from 2001 to 2002. Furthermore, L. pneumophila serogroup 3 with an identical PFGE profile B was identified in 20 environmental strains from all wards, L. pneumophila serogroup 3 with PFGE profile C was identified in a single environmental strain from the CR-ICU, and non-pneumophila Legionella with identical PFGE profile D was identified in five environmental strains from the adult CSU, paediatric CSU and NICU. Ultraviolet irradiation was effective in disinfection of the hospital water supplies in the adult and paediatric CSUs contaminated by L. pneumophila clone associated with nosocomial Legionnaires' disease. In conclusion, these data demonstrate that two cases of nosocomial legionellosis were caused by the persistence of a single clone of L. pneumophila serogroup 1/Philadelphia in the hospital environment, and that disinfection by ultraviolet irradiation may represent an effective measure to prevent nosocomial Legionnaires' disease. © 2005 The Hospital Infection Society. Published by Elsevier Ltd. All rights reserved
Histochemical analysis of glycoconjugates in the domestic cat testis
The localization and characterization of
oligosaccharide sequences in the cat testis was
investigated using 12 lectins in combination with the Ăelimination
reaction, N-Glycosidase F and sialidase
digestion. Leydig cells expressed O-linked glycans with
terminal aGalNAc (HPA reactivity) and N-glycans with
terminal/internal aMan (Con A affinity). The basement
membrane showed terminal Neu5Aca2,6Gal/GalNAc,
GalĂ1,3GalNAc, a/ĂGalNAc, and GlcNAc (SNA, PNA,
HPA, SBA, GSA II reactivity) in O-linked oligosaccharides,
terminal GalĂ1,4GlcNAc (RCA120 staining)
and aMan in N-linked oligosaccharides; in addition,
terminal Neu5Aca2,3GalĂ1,4GlcNac, Forssman
pentasaccharide, aGal, aL-Fuc and internal GlcNAc
(MAL II, DBA, GSA I-B4, UEA I, KOH-sialidase-WGA
affinity) formed both O- and N-linked oligosaccharides.
The Sertoli cells cytoplasm contained terminal Neu5Ac-
GalĂ1,4GlcNAc, Neu5Ac-ĂGalNAc as well as internal
GlcNAc in O-linked glycans, aMan in N-linked
glycoproteins and terminal Neu5Aca2,6Gal/
GalNAc in both O- and N-linked oligosaccharides.
Spermatogonia exhibited cytoplasmic N-linked
glycoproteins with aMan residues. The spermatocytes
cytoplasm expressed terminal Neu5Aca2,3GalĂ1,4
GlcNAc and GalĂ1,3GalNAc in O-linked oligosaccharides,
terminal GalĂ1,4GlcNAc and a/ĂGalNAc
in N-linked glycoconjugates. The Golgi region showed
terminal Neu5aca2,3GalĂ1,4GlcNac, GalĂ1,4GlcNAc,
Forssman pentasaccharide, and aGalNAc in O-linked
oligosaccharides, aMan and terminal ĂGal in N-linked
oligosaccharides. The acrosomes of Golgi-phase
spermatids expressed terminal GalĂ1,3GalNAc,
GalĂ1,4GlcNAc, Forssmann pentasaccharide,
a/ĂGalNAc, aGal and internal GlcNAc in O-linked
oligosaccharides, terminal a/ĂGalNAc, aGal and
terminal/internal aMan in N-linked glycoproteins. The
acrosomes of cap-phase spermatids lacked internal
Forssman pentasaccharide and aGal, while having
increased a/ĂGalNAc. The acrosomes of elongated
spermatids did not show terminal GalĂ1,3GalNAc,
displayed terminal GalĂ1,4GlcNAc and a/ĂGalNAc in
N-glycans and Neu5Ac-GalĂ1,3GalNAc in O-linked
oligosaccharides
Molecular Epidemiology and Virulence Profiles of Colistin-Resistant Klebsiella pneumoniae Blood Isolates From the Hospital Agency âOspedale dei Colli,â Naples, Italy
Resistance to colistin is increasingly reported in Klebsiella pneumoniae clinical isolates. The aim of this study was to analyze the molecular epidemiology and virulence profiles of 25 colistin-resistant K. pneumoniae blood isolates from the Hospital Agency âOspedale dei Colli,â Naples, Italy, during 2015 and 2016. Colistin MIC values of isolates ranged from 4 to 256 mg/L. The inactivation of the mgrB gene, encoding a negative regulator of the PhoQ/PhoP signaling system, was the most frequent mechanism of colistin resistance found in 22 out of 25 isolates. Of these, 10 isolates assigned to ST512 and PFGE types A and A4 showed identical frameshift mutation and premature termination of mgrB gene; 4 isolates assigned to ST258 and PFGE types A1 showed non-sense, frameshift mutation, and premature termination; 3 and 1 isolates assigned to ST258 and PFGE A2 and ST512 and PFGE A3, respectively, had insertional inactivation of mgrB gene due to IS5-like mobile element; 2 isolates assigned to ST101 and 1 to ST392 had missense mutations in the mgrB gene, 1 isolate assigned to ST45 showed insertional inactivation of mgrB gene due to IS903-like mobile element. phoQ missense mutations were found in 2 isolates assigned to ST629 and ST101, respectively, which also showed a missense mutation in pmrA gene. The mcr-1-2-3-4 genes were not detected in any isolate. Colistin-resistant K. pneumoniae isolates showed variable virulence profiles in Galleria mellonella infection assays, with the infectivity of two isolates assigned to ST45 and ST629 being significantly higher than that of all other strains (P < 0.001). Interestingly, colistin MIC values proved to make a significant contribution at predicting lethal doses values (LD50 and LD90) of studied isolates in G. mellonella. Our data show that MgrB inactivation is a common mechanism of colistin resistance among K. pneumoniae in our clinical setting. The presence of identical mutations/insertions in isolates of the same ST and PFGE profile suggests the occurrence of clonal expansion and cross-transmission. Although virulence profiles differ among isolates irrespective of their genotypes, our results suggest that high colistin MIC could predict lower infectivity capability of the isolates
Physical activity regulates tnfα and il-6 expression to counteract inflammation in cystic fibrosis patients
Background: Cystic fibrosis (CF) is one of the most common inherited diseases. It is characterised by a severe decline in pulmonary function associated with metabolic perturbations and an increased production of inflammatory cytokines. The key role of physical activity (PA) in improving the health status of CF patients and reducing lung function decline has recently been demonstrated. This study evaluated interleukin-6 (IL-6) and tumour necrosis factor α (TNFα) expression in two subgroups of CF patients classified based on PA. Methods: We selected 85 CF patients; half of them regularly undertook supervised PA in the three years leading up to the study and half of them were not physically active. Patients were analysed for serum IL-6 and TNFα levels using enzyme-linked immunosorbent assays. Results: We found that the expression levels of IL-6 and TNFα differed in terms of their regulation by PA. In particular, TNFα levels negatively correlated with FEV1% decrease/year and FEV1% decrease (p = 0.023 and p = 0.02, respectively), and positively correlated with serum fasting glucose (p = 0.019) in PA CF patients. In contrast, in the NPA subgroup, TNFα levels were positively correlated with IL-6 (p = 0.001) and negatively correlated with adiponectin (p = 0.000). In addition, multiple logistic regression analysis confirmed that PA is an independent modulator of the inflammatory state. Conclusions: PA modulates inflammatory processes in CF patients by regulating the secretion of pro-inflammatory cytokines and thus ameliorating lung function. Our data show that PA is a useful complementary strategy in the management of CF and that TNFα may be a marker of these effects of PA
A novel IncA/C1 group conjugative plasmid, encoding VIM-1 metallo-beta-lactamase, mediates the acquisition of carbapenem resistance in ST104 Klebsiella pneumoniae isolates from neonates in the intensive care unit of V. Monaldi Hospital in Naples
The emergence of carbapenemase producing Enterobacteriaceae has raised major public health concern. The aim of this study was to investigate the molecular epidemiology and the mechanism of carbapenem resistance acquisition of multidrug-resistant Klebsiella pneumoniae isolates from 20 neonates in the neonatal intensive care unit (NICU) of the V. Monaldi Hospital in Naples, Italy, from April 2015 to March 2016. Genotype analysis by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) identified PFGE type A and subtypes A1 and A2 in 17, 2, and 1 isolates, respectively, and assigned all isolates to sequence type (ST) 104. K. pneumoniae isolates were resistant to all classes of ÎČ-lactams including carbapenems, fosfomycin, gentamicin, and trimethoprim-sulfamethoxazole, but susceptible to quinolones, amikacin, and colistin. Conjugation experiments demonstrated that resistance to third-generation cephems and imipenem could be transferred along with an IncA/C plasmid containing the extended spectrum ÎČ-lactamase blaSHV -12 and carbapenem-hydrolyzing metallo-ÎČ-lactamase blaV IM-1 genes. The plasmid that we called pIncAC_KP4898 was 156,252 bp in size and included a typical IncA/C backbone, which was assigned to ST12 and core genome (cg) ST12.1 using the IncA/C plasmid MLST (PMLST) scheme. pIncAC_KP4898 showed a mosaic structure with blaV IM-1 into a class I integron, blaSHV -12 flanked by IS6 elements, a mercury resistance and a macrolide 2'-phosphotransferase clusters, ant(3âł), aph(3âł), aacA4, qnrA1, sul1, and dfrA14 conferring resistance to aminoglycosides, quinolones, sulfonamides, and trimethoprim, respectively, several genes predicted to encode transfer functions and proteins involved in DNA transposition. The acquisition of pIncAC_KP4898 carrying blaV IM-1 and blaSHV -12 contributed to the spread of ST104 K. pneumoniae in the NICU of V. Monaldi Hospital in Naples
Robust Model Predictive Control of An Input Delayed Functional Electrical Stimulation
Functional electrical stimulation (FES) is an external application of low-level currents to elicit muscle contractions that can potentially restore limb function in persons with spinal cord injury. However, FES often leads to the rapid onset of muscle fatigue, which limits performance of FES-based devices due to reduction in force generation capability. Fatigue is caused by unnatural muscle recruitment and synchronous and repetitive recruitment of muscle fibers. In this situation, overstimulation of the muscle fibers further aggravates the muscle fatigue. Therefore, a motivation exists to use optimal controls that minimize muscle stimulation while providing a desired performance. Model predictive controller (MPC) is one such optimal control method. However, the traditional MPC is dependent on exact model knowledge of the musculoskeletal dynamics and cannot handle modeling uncertainties. Motivated to address modeling uncertainties, robust MPC approach is used to control FES. Moreover, two new robust MPC techniques are studied to address electromechanical delay (EMD) during FES, which often causes performance issues and stability problems.
This thesis compares two types of robust MPCs: a Lyapunov-based MPC and a tube- based MPC for controlling knee extension elicited through FES.
Lyapunov-based MPC incorporated a contractive constraint that bounds the Lyapunov function of the MPC with a Lyapunov function that was used to derive an EMD compensation control law. The Lyapunov-based MPC was simulated to validate its performance.
In the tube-based MPC, the EMD compensation controller was chosen to be the tube that eliminated output of the nominal MPC and the output of the real system. Regulation experiments were performed for the tube-based MPC on a leg extension machine and the controller showed robust performance despite modeling uncertainties
An Ethnographer Lured into Darkness
No matter the combination of methods ethnographers bring to their research design and to participant observation, our pursuit to log, interpret, analyse and present the lives of those we meet is never an entirely intellectual or objective one. Ethnographic fieldwork is intimately sensory (Pink, Doing sensory ethnography, Sage, London, 2015), invokes our imagination (Sparkes, Qualitative research in sport and exercise, 1:21â35, 2009) and requires us to actively navigate social landscapes (Hammersley and Atkinson, Field relations. Ethnography: Principles in practice, Routledge, Stoodleigh, 2007). There is a tendency for these elements to fade in terms of visibility and immediacy within the research process. For those in accord with (Davies, Reflexive ethnography: A guide to researching selves and others, Routledge, New York, 2008), continuous reflexive labour becomes a core praxis to monitor the ways we observe and participate in this textured environment. Without this, we are left in the dark and are less able to see how we can (or should) respond to the nittyâgritty qualitative nature of ethnography. In this Chapter, two of methodological vignettes will act as entry points to unpack a set of tensions that commanded my attention during an eighteen months ethnography in Higher Education. âYou Look Like an Ivory Tower Studentâ, for example, begins to troubleshoot ethnographic participation within educational environments. âGoing Darkâ, on the other hand, problematises the prioritisation of visual observations that are implicit in ethnographic tradition. Throughout these discussions a metaphor of being lured into darkness is offered as a productive orientation for ethnography
The RING network: improvements to a GPS velocity field in the central Mediterranean
Since 2004, a continuous Global Positioning System (GPS) network has been
operated by the Istituto Nazionale di Geofisica e Vulcanologia (INGV) to
investigate active tectonic processes in Italy and the surrounding regions,
which are still largely debated. This important infrastructure is known as
Rete Integrata Nazionale GPS (RING) network, and it consists of about 130
stations that are deployed all over Italy. The development and realization of
a stable GPS monumentation, its integration with seismological instruments,
and the choice of both satellite and internet data transmission, make this
network one of the most innovative and reliable CGPS networks in the
world. The technologically advanced development of the RING network has
been accompanied by the development of different data processing strategies,
which are mainly dependent on the use of different GPS analysis software.
The different software-related solutions are here compared at different scales
for this large network, and the consistency is evaluated and quantified within
an RMS value of 0.3 mm/yr
- âŠ