Azithromycin regulates bacterial virulence and immune response in a murine model of ceftazidime-treated Pseudomonas aeruginosa acute pneumonia.

Pseudomonas aeruginosa (PA) remains one of the leading causes of nosocomial acute pneumonia. The array of virulence factors expressed by PA and the intense immune response associated with PA pneumonia play a major role in the severity of these infections. New therapeutic approaches are needed to overcome the high resistance of PA to antibiotics and to reduce the direct damage to host tissues. Through its immunomodulatory and anti-virulence effects, azithromycin (AZM) has demonstrated clinical benefits in patients with chronic PA respiratory infections. However, there is relatively little evidence in PA acute pneumonia. We investigated the effects of AZM, as an adjunctive therapy combined with ceftazidime (CAZ), in a murine model of PA acute pneumonia. We observed that the combined therapy (i) reduces the weight loss of mice 24 h post-infection (hpi), (ii) decreases neutrophil influx into the lungs at 6 and 24 hpi, while this effect is absent in a LPS-induced pneumonia or when PA is pretreated with antibiotics and mice do not receive any antibiotics, and that (iii) AZM, alone or with CAZ, modulates the expression of PA quorum sensing regulators and virulence factors (LasI, LasA, PqsE, PhzM, ExoS). Our findings support beneficial effects of AZM with CAZ on PA acute pneumonia by both bacterial virulence and immune response modulations. Further investigations are needed to clarify the exact underlying mechanisms responsible for the reduction of the neutrophils influx and to better discriminate between direct immunomodulatory properties of AZM, and indirect effects on neutrophilia resulting from bacterial virulence modulation.

Prevalence of a cefazolin inoculum effect associated with blaZ gene types, and clinical outcomes among methicillin-susceptible Staphylococcus aureus blood isolates of patients with infective endocarditis.

OBJECTIVES: A proportion of blaZ gene-positive methicillin-susceptible Staphylococcus aureus (MSSA) strains exhibits the cefazolin inoculum effect (CInE). Its clinical impact remains uncertain but could compromise the use of cefazolin in high-burden infections. To date, no study has been conducted in France or in Europe. We aimed to assess the prevalence of CInE and its association with blaZ beta-lactamase and S. aureus protein A (spa) types, and to assess the clinical outcomes in cefazolin-treated patients for infective endocarditis whose strain exhibited a CInE. METHODS: This was a French single-center retrospective study of 51 MSSA strains from patients of the Nantes endocarditis prospective cohort, conducted between 2013 and 2018. RESULTS: Cefazolin MIC50 at high inoculum was 2 mg/L (IQR 1-2). CInE was found in 17.6 % of tested strains. Among blaZ-positive strains (n = 44), type A beta-lactamase was predominant (n = 25, 57 %). Thirty-seven S. aureus protein A (spa) types were found. No statistical association was shown between blaZ or spa types and CInE. CInE was neither associated with a higher rate of persistent bacteremia (25 % vs 56.3 %, p = 0.58) nor with clinical failure in patients treated with cefazolin, in comparison to patients with no CInE strain (25 % vs 56.3 %, p = 0.58). CONCLUSION: The cefazolin inoculum effect was found in a substantial number of Staphylococcus aureus strains; however, minimum inhibitory concentrations remained globally low. CInE was not associated with a higher proportion of clinical failure on treatment.

Innate immune evasion of Escherichia coli clinical strains from orthopedic implant infections.

Escherichia coli is one of the first causes of Gram-negative orthopedic implant infections (OII). Those infections, usually hematogenous, mostly originate from the urinary tract. We investigated the strategies developed by E. coli in this context to evade host innate immune responses, i.e. complement and polymorphonuclear neutrophils (PMN). Twenty strains from OII were compared with 20 strains from bacteremia in patients with non-infected orthopedic implant. In both groups, 6/20 (30 %) strains lysed PMNs, due to the production of the pore-forming toxin α-hemolysin (HlyA). For the others, resistance to phagocytic killing by PMN was not significantly different between both groups. In contrast, resistance to complement-mediated serum killing was significantly higher in OII strains than in the others (65 % vs 10 %; P <0.001). In E. coli, different mechanisms have been involved in complement resistance. Here, serum resistance was not linked to a group 2 capsule, or a loss of outer membrane permeability, or the recruitment of the complement inhibitor C4bp, but was significantly associated with the synthesis of long-chain LPS, regardless of the O-antigen. Thus, serum resistance could promote seeding of peri-implant tissues by helping E. coli to either persist in blood and reach the site of infection or overcome localized complement activation.

Molecular mechanisms involved in the antiproliferative effect of two COX-2 inhibitors, nimesulide and NS-398, on colorectal cancer cell lines.

BACKGROUND: Cyclooxygenase (COX)-2 is up-regulated in most colorectal cancers. Chronic use of non-steroidal anti-inflammatory drugs, which target cyclooxygenases, have been shown to reduce the risk of these cancers. However, the mechanisms underlying this protective effect remain unclear. AIMS: The aim of our study was to characterize the effects of two COX-2 selective inhibitors, NS-398 and nimesulide, on colorectal cancer cell proliferation, and to describe the molecular mechanisms involved. MATERIALS AND METHODS: HT-29 and SW-1116 cell lines were cultured with either NS-398 or nimesulide. Cell proliferation was assessed by staining DNA with crystal violet. Cell cycle repartition and apoptosis were analysed by flow cytometry. The expression of COX-1 and COX-2. and of two cyclin dependent kinase inhibitors, p21Cip1 and p27Kip1, was analysed by Western blotting and RT-PCR. RESULTS: Both drugs dose-dependently inhibited cell proliferation and induced G1 cell cycle blockade. HT-29 cells were more sensitive to both drugs than SW-1116 cells. p21Cip1 and p27Kip1 were induced on both cell lines. Concomitant induction of p21Cip1 mRNA indicates transcriptional modulation, whereas induction of p27Kip1 only at the protein level suggests post-translational modulation. CONCLUSION: NS-398 and nimesulide inhibit colorectal cell proliferation through induction of p21Cip1 and p27Kip1.

Home environmental consequences of commute travel impedance.

The physical and perceptual dimensions of commuting travel impedance were again found to have stressful consequences in a study of 99 employees of two companies. This quasi-experimental replication study, which focuses here on home environment consequences, investigated the effects of physical impedance and subjective impedance on multivariate measures of residential satisfaction and personal affect in the home. Both sets of residential outcome measures were significantly related to the two impedance dimensions. As predicted, gender was a significant moderator of physical impedance effects. Women commuting on high physical impedance routes were most negatively affected. Previously found subjective impedance effects on negative home mood, regardless of gender, were strongly replicated with several methods and were buttressed by convergent results with objective indices. The theoretical conjecture that subjective impedance mediates the stress effects of physical impedance was supported by the personal affect cluster but only for one variable in the residential satisfaction cluster. Traffic congestion has increased in metropolitan areas nationwide, and commuters, families, and organizations are absorbing associated hidden costs. The results are reviewed in terms of our ecological model, and the moderating effects of gender are discussed in terms of choice and role constraints.

Developmental and gender differences in anonymous support-seeking: analysis of data from a community help line for children.

Calls to a community help line for children were analyzed for developmental and gender trends in support-seeking during middle childhood. Results indicated that between the ages of 10 and 11 the nature of help-seeking by callers changed, with a shift toward more informational support-seeking and concern regarding interpersonal problems. This shift was particularly dramatic for girls. Gender differences in the type of support sought (informational vs. instrumental) were slight. Parallels with other studies of stress and coping in children are drawn.