A bacterial biosensor encoding a genetically modified LuxR receptor exhibits improved detection of Pseudomonas aeruginosa's biomarker molecule 2-aminoacetophenone.

2-Aminoacetophneone (2-AA) is a volatile molecule produced in high amounts by the opportunistic pathogen Pseudomonas aeruginosa. We have previously shown that 2-AA activates the quorum sensing (QS) LuxR receptor of Aliivibrio fischeri. In the present study we were able to improve LuxR's affinity and detection limit for 2-AA by genetic modification of three amino acids within the binding pocket of the receptor. Expression of the modified LuxR receptor in a luminescent bacterial biosensor provided an efficient detection assay of 2-AA in clinical P. aeruginosa strains isolated from blood and lung infections, as well as in phlegm samples obtained from subjects suffering from lung infections.

Staffing for infectious diseases, clinical microbiology and infection control in hospitals in 2015: results of an ESCMID member survey.

We aimed to assess the current status of infectious diseases (ID), clinical microbiology (CM) and infection control (IC) staffing in hospitals and to analyse modifiers of staffing levels. We conducted an Internet-based survey of European Society of Clinical Microbiology and Infectious Diseases members and affiliates, collecting data on hospital characteristics, ID management infrastructure, ID/IC-related activities and the ratio of physicians per 100 hospital beds. Regression analyses were conducted to examine factors associated with the physician-bed ratio. Five hundred sixty-seven hospital responses were collected between April and June 2015 from 61 countries, 81.2% (384/473) from Europe. A specialized inpatient ward for ID patients was reported in 58.4% (317/543) of hospitals. Rates of antibiotic stewardship programmes (ASP) and surveillance activities in survey hospitals were high, ranging from 88% to 90% for local antibiotic guidelines and 70% to 82% for programmes monitoring hospital-acquired infections. The median ID/CM/IC physician per 100 hospital beds ratio was 1.12 (interquartile range 0.56-2.13). In hospitals performing basic ASP and IC (including local antibiotic guidelines and monitoring device-related or surgical site infections), the ratio was 1.21 (interquartile range 0.57-2.14). Factors independently associated with higher ratios included compliance with European Union of Medical Specialists standards, smaller hospital size, tertiary-care institution, presence of a travel clinic, beds dedicated to ID and a CM unit. More than half of respondents estimated that additional staffing is needed for appropriate IC or ID management. No standard of physician staffing for ID/CM/IC in hospitals is available. A ratio of 1.21/100 beds will serve as an informed point of reference enabling ASP and infection surveillance.

Increased rates of intensive care unit admission in patients with Mycoplasma pneumoniae: a retrospective study.

Mycoplasma pneumoniae is a leading cause of respiratory disease. In the Intensive Care Unit (ICU) setting M. pneumoniae is not considered a common pathogen. In 2010-13 an epidemic of M. pneumoniae-associated infections was reported and we observed an increase of M. pneumoniae patients admitted to ICU. We analysed the cohort of all M. pneumoniae-positive patients' admissions during 2007 to 2012 at the Hadassah-Hebrew University Medical Centre (a 1100-bed tertiary medical centre). Mycoplasma pneumoniae diagnosis was made routinely using PCR on throat swabs and other respiratory samples. Clinical parameters were retrospectively extracted. We identified 416 M. pneumoniae-infected patients; of which 68 (16.3%) were admitted to ICU. Of these, 48% (173/416) were paediatric patients with ICU admission rate of 4.6% (8/173). In the 19- to 65-year age group ICU admission rate rose to 18% (32/171), and to 38.8% (28/72) for patients older than 65 years. The mean APACHE II score on ICU admission was 20, with a median ICU stay of 7 days, and median hospital stay of 11.5 days. Of the ICU-admitted patients, 54.4% (37/68) were mechanically ventilated upon ICU admission. In 38.2% (26/68), additional pathogens were identified mostly later as secondary pathogens. A concomitant cardiac manifestation occurred in up to 36.8% (25/68) of patients. The in-hospital mortality was 29.4% (20/68) and correlated with APACHE II score. Contrary to previous reports, a substantial proportion (16.3%) of our M. pneumoniae-infected patients required ICU admission, especially in the adult population, with significant morbidity and mortality.

Exposure to ertapenem is possibly associated with Pseudomonas aeruginosa antibiotic resistance.

The role of antibiotic exposure in the evolution and emergence of resistance is challenging to assess. We used carbapenem-resistant Pseudomonas aeruginosa (PA) phenotypes to assess possible factors that are associated with the occurrence and prognosis of such a phenotype and to examine the possible contribution of antibiotic exposure to the evolution of antimicrobial resistance. We conducted a nested case-control study. Cases were defined as patients from whom carbapenem-resistant ureidopenicillin-sensitive PA (CRUS-PA) was isolated; matched controls were PA patients who did not have isolation of CRUS-PA. We analysed potential predictors of CRUS-PA isolation and assessed their clinical significance (mortality and eventual isolation of pan-resistant PA), taking into account antibiotic exposures. We matched 800 case-control pairs. Case patients were more likely to have been exposed to anti-PA carbapenems (OR = 6.9; 95% CI, 2.5-18.6). This finding did not apply to the administration of other antibiotics. The mortality among CRUS-PA patients was similar to that of the controls (HR, 0.8 95%; CI, 0.6-1.1). Subsequent isolation of pan-resistant PA was more frequent among case patients compared with non-pan-resistant controls (p-value <0.05). Among cases, the risk of eventual pan-resistant PA isolation was increased in ertapenem recipients, only after and not prior to the index specimen date (HR, 1.9, 95%; CI, 1.01-3.4). Therefore we suggest that the CRUS-PA phenotype may represent pan beta-lactam resistance and that antibiotic exposure is associated with evolution of PA resistance phenotypes. We demonstrate a novel association of ertapenem with sequentially appearing PA resistance patterns.

Ongoing epidemic of Mycoplasma pneumoniae infection in Jerusalem, Israel, 2010 to 2012.

A substantial epidemic of Mycoplasma pneumonia infection was reported in late 2011 in some European countries. We report here an epidemic of M. pneumonia infection that began in Jerusalem during 2010 and is still ongoing. This report complements current information on what might be a worldwide epidemic of M. pneumoniae infection that might require substantial coordinated international public health intervention.

Streptococcus pyogenes emm and T types within a decade, 1996-2005: implications for epidemiology and future vaccines.

Streptococcus pyogenes group A (GAS) is a primary human pathogen. We performed genetic emm sequence and serological T-antigen typing of 819 mostly invasive GAS isolates recovered in Israel during 1996-2005. Of the 72 emm types found, the six most prevalent types (1, 81, 89, 14, 28, 5) comprised 30.2% of all isolates, and emm-type changes were observed over the years. The predicted coverage of the 26-valent S. pyogenes vaccine formulated for usage in the USA was predicted to be only approximately 60%. On the basis of different emm-T antigen type associations, some Israeli strains are probably different clonal types than those found in USA. About 2% of GAS had emm types that were originally associated with S. dysgalactiae subsp. equisimilis emm genes. Therefore, routine emm typing allows meaningful GAS strain surveillance, and provides data relevant to better vaccine coverage.

Acute otitis media in the first two months of life: characteristics and diagnostic difficulties.

OBJECTIVE: To assess the clinical and laboratory features of acute otitis media (AOM) in infants younger than 2 months, to look for factors predicting bacterial otitis, and to evaluate the accuracy of AOM diagnosis among paediatricians. METHODS: The study population comprised a cohort of 277 hospitalised infants up to 61 days old that were treated for the first episode of AOM in a paediatric department. We reviewed their medical records and analysed the demographic, clinical and laboratory data, and the diagnosis made by both paediatricians and otolaryngologists. RESULTS: Presenting symptoms were mainly respiratory (70.0%) and fever (62.5%). The most common pathogens were Streptococcus pneumoniae and Haemophilus influenzae. Gram-negative bacilli grew in 10.5% of the infants. Multivariate analysis revealed that AOM in the second month of life was associated with male gender, concurrent bronchiolitis and diarrhea. Although high leukocyte count was associated with bacterial pathogen, more than 70% of the patients with positive culture had normal white blood cell counts. The paediatrician diagnosed only 45% of the patients subsequently diagnosed with AOM by an otolaryngologist. CONCLUSIONS: The absence of predictors for bacterial infection in more than 70% of bacterial AOM suggests that empirical antibiotic treatment should be advised for the young infants with AOM even when afebrile and with normal laboratory profile. A low diagnostic rate of AOM by the paediatrician emphasizes the need for improvement in examination skills and instrumentation to allow a thorough ear evaluation in children of a very young age.

Evaluation of eight commercial tests for Mycoplasma pneumoniae antibodies in the absence of acute infection.

Eight commercially available tests for Mycoplasma pneumoniae (Serodia-Myco II, Labsystems IgM and IgG EIA, IgM and IgG LISA tests, ImmunoWell IgG test and SeroMP IgM and IgG) were compared using 204 single sera from healthy individuals. IgM peaked in late childhood and then declined, while IgG rose progressively into adulthood. Inter-assay agreement was poor. Positivity in Serodia-Myco II and LISA IgG was associated with blood group or Coombs positivity, suggesting non-specific reactions. The study confirmed that single serum serology is unsuitable for the diagnosis of M. pneumoniae infection, and that commercially available tests need further improvement.

Severe influenza infection in a chronic hepatitis C carrier: failure of protective serum HI antibodies after IM vaccination.

BACKGROUND: Influenza is an important cause of morbidity and mortality in immunocompromised hosts. Recommendations exists for vaccination each year, yet disease can still occur. OBJECTIVES: To describe the course of fulminant influenza infection in a patient with HCV. STUDY DESIGN: Case study in which correlation was made between immunoglobulin response to influenza vaccination to the disease and its unique clinical course caused by influenza virus. RESULTS: Influenza A/Jerusalem 17/98 (H(1)N(1)) was isolated from the throat of a chronic hepatitis C carrier who, presented with shortness of breath, and subsequent massive bilateral pneumonia. The patient was previously immunized IM with inactive influenza vaccine. He developed protective levels of humoral antibodies (1:80 hemagglutination inhibition (HI) antibodies) against the three strains of the vaccine that evidently did not prevent respiratory infection. The development of massive bilateral pneumonia and continued presence of influenza virus in the respiratory tract may have been due to his underlying medical condition and possible lack of mucosal secretory IgA (SIgA) antibodies. CONCLUSION: We have presented a case of prolonged influenza infection post vaccination. This case emphasizes the importance of an improved vaccine that would stimulate a better immunologic response, especially in immunocompromised patients.

Reduction of jaw opening (trismus) in giant cell arteritis.

OBJECTIVE: To study the prevalence and the clinical characterisation of jaw problems in patients with giant cell arteritis (GCA). METHODS: the prevalence of such symptoms in patients with GCA was evaluated by performing a retrospective analysis of all patients with GCA and polymyalgia rheumatica who were diagnosed during admission to Hadassah University Hospital. Ten patients reported previously in the literature were also evaluated. RESULTS: Six patients out of 88 (6.8%) had complaints of reduction in jaw opening. These six patients seemed to have a much more abrupt onset of disease with shorter duration until diagnosis, higher prevalence of eye involvement (50% v 27%), and a higher rate of positive pathology (100%). CONCLUSIONS: Reduction in jaw opening in the appropriate setting may indicate the presence of GCA. This sign should not be overlooked in the presence of the claudication sign as it seems to reflect more severe GCA disease.

Compound cardiac toxicity of oral erythromycin and verapamil.

OBJECTIVE: To report a case of complete atrioventricular (AV) block and QTc prolongation following coadministration of high-dose verapamil and erythromycin. CASE SUMMARY: A 79-year-old white woman was admitted to the hospital due to extreme fatigue and dizziness. On admission, heart rate was 40 beats/min and blood pressure was 80/40 mm Hg. An electrocardiogram showed complete atrioventricular (AV) block, escape rhythm of 50 beats/min, and QTc prolongation 583 msec. This event was attributed to concomitant treatment with verapamil 480 mg/d and erythromycin 2,000 mg/d, which was prescribed one week before admission. DISCUSSION: This is the first case published describing complete AV block and prolongation of QTc following coadministration of erythromycin and verapamil. CYP3A4 is the main isoenzyme responsible for metabolism of erythromycin and verapamil. Both drugs are potent inhibitors of CYP3A4 and of P-glycoprotein; this may be the basis for the pharmacokinetic interaction between erythromycin and verapamil. In addition to being a woman, our patient had other risk factors for QT prolongation: slow baseline heart rate (probably induced by verapamil), left-ventricular hypertrophy, and possibly ischemic heart disease. CONCLUSIONS: This life-threatening arrhythmia was probably the result of a pharmacokinetic and/or pharmacodynamic interaction of high-dose verapamil and erythromycin.

Unusual presentation of familial Mediterranean fever: role of genetic diagnosis.

OBJECTIVE: To describe the role of molecular analysis in the diagnosis of an unusual presentation of familial Mediterranean fever (FMF). CASE REPORT: Two patients presenting with prolonged fever without signs and symptoms of serositis are described. FMF was diagnosed by genetic analysis, which disclosed that both patients were homozygous for the M694V mutation of the Mediterranean fever (MEFV) gene. CONCLUSION: Molecular analysis of FMF should complement the investigation of patients with fever of unknown origin. This test enables a definite diagnosis of the disease and may promote the diagnosis and treatment of patients with an unusual or incomplete clinical picture of FMF.

Acute hyperphosphatemia caused by sodium phosphate enema in a patient with liver dysfunction and chronic renal failure.

We report a case of acute hyperphosphatemia secondary to rectal administration of sodium phosphate and sodium biphosphate (Fleet enema). Parathyroid hormone and calcitonin levels were measured along with phosphate clearance and the tubular reabsorption of phosphate.

Factors predicting mortality of patients with lung abscess.

BACKGROUND: The rates of morbidity and mortality associated with lung abscess are still significant despite the introduction of antibiotic treatments. The aim of this work was to identify the factors that predict a poor outcome for patients with lung abscess. METHODS: We retrospectively reviewed the records and the roentgenographic files of adult patients with lung abscess who were hospitalized from 1980 to 1996 at the Hadassah University Hospital, in Jerusalem, Israel. RESULTS: The study population comprised 75 patients, and the mean age was 52 years old (range, 12 to 89 years). The mean (+/- SD) hospitalization duration was 25.7+/-21.5 days (range, 5 to 94 days). Fifteen patients (20%) succumbed to the infection. The patients who died had more predisposing factors (+/-SD), such as pneumonia, neoplasm, and altered consciousness, than those who survived, respectively: 2.73+/-1.4 vs 1.9+/-1.3 (p < 0.03). The patients with anemia on admission (hemoglobin levels of < 10 g/dL) had a higher mortality rate than those with higher hemoglobin levels, respectively: 58.3 vs 12.9% (p = 0.0008). A higher mortality rate was also associated with infection by Pseudomonas aeruginosa (83%), Staphylococcus aureus (50%), and Klebsiella pneumoniae (44%). The patients who died had larger abscess volumes (+/-SD) than those who survived (233+/-99 vs 157+/-33 mL), although it did not reach statistical significance. The diameter of the abscess correlated with the hospitalization time (r = 0.5; p < 0.001). CONCLUSION: High rates of morbidity and mortality are associated with lung abscess despite appropriate antibiotic therapy and better supportive care. In patients with several predisposing factors, such as a large abscess size and a right-lower-lobe location, the prognosis was worse. The patients infected with S aureus, K pneumoniae, and particularly P aeruginosa had an ominous prognosis. As the prognosis for lung abscess has not improved sufficiently since the introduction of antibiotics, other modalities should be considered for patients with poor prognostic signs.

Mycoplasmas regulate HIV-LTR-dependent gene expression.

Mycoplasmas have been incriminated in setting the stage for HIV infection and full-blown AIDS. We tested the possible involvement of mycoplasmas in activation of HIV. Two cell lines, 293 fibroblasts and Jurkat CD4+ T-cells, transfected with plasmids harboring a transcription fusion construct between HIV-long terminal repeat (HIV-LTR) and either luc or cat genes, were infected with several mycoplasmas (M. fermentans; M. penetrans, M. pirum and Ureaplasma urealyticum) and the reporter gene expression was monitored. The data presented here suggest that mycoplasmas, and specifically their membranes, play a role in the activation of HIV-LTR mediated transcription.