Predominance of ST8 and CC1/spa-t1784 methicillin-resistant Staphylococcus aureus isolates in Japan and their genomic characteristics.

OBJECTIVES: Methicillin-resistant Staphylococcus aureus (MRSA) has become a serious epidemiologic problem worldwide. In this study, we aimed to investigate recently isolated MRSA types and determine their characteristics. METHODS: We collected 164 strains isolated from 13 hospitals located in Tokyo and surrounding prefectures. In addition to drug resistance tests, we sequenced whole genomes of the prevalent MRSA clones and analysed their genomic characteristics, such as drug resistance genes, virulence factor genes, and genome arrangements. RESULTS: Multilocus sequencing typing showed that 51% of the SCCmecⅣ MRSA isolates belonged to clonal complex 1 (CC1). Staphylococcus protein A gene (spa) typing showed that 91% of these CC1 isolates could be categorised as t1784 type. These CC1/t1784 isolates possessed genes encoding erythromycin resistance protein, spectinomycin 9-adenylyltransferase, and staphylococcal enterotoxins (SEA, SEI, SEM), but not the pvl gene encoding Panton-Valentine leukocidin. Complete genomic analysis of nine CC1/t1784 isolates showed that they shared an intact phage, which carried no annotated virulence factor genes except for two encoding a hypothetical membrane protein and a teichoic acid biosynthesis protein. No significant genomic rearrangements were observed among the CC1/t1784 isolates. CONCLUSION: These data and previous reports indicate that this CC1/t1784 clone has been expanding rapidly in Japan without genomic changes.

Rapid detection of bacteria that produce extended-spectrum ß-lactamase by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

OBJECTIVES: Proper use of antibacterial agents is necessary to prevent the spread of drug-resistant bacteria. To support clinicians, laboratories need to rapidly determine bacterial drug susceptibility/resistance. We have established a method to distinguish extended-spectrum ß-lactamase (ESBL)-producing clinical isolates by capturing structural changes in ß-lactam antibiotics using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). METHODS: Clinical isolates of Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis, classified into ESBL-producing strains and sensitive strains based on the presence or absence of a CTX-M-type gene, were used. Test bacteria were cultured aerobically in solid-phase wells of Eiken DPD1 dry plates at 35°C for 15 min or 30 min with the antibiotics cefotaxime (CTX), cefpodoxime (CPDX) or piperacillin (PIPC). Culture supernatants were then used for analysis with a MALDI Biotyper. RESULTS: Signals derived from non-hydrolyzed products of antibiotics were observed in all strains. In the case of ESBL-producing strains, signals derived from the hydrolysis products of antibiotics were also observed. Since the ratio of signal intensity derived from hydrolysis products divided by the total signal intensity detected was ≥11% for CTX and ≥6% for CPDX and PIPC, all strains were determined to be ESBL-producing bacteria. CONCLUSION: The short incubation time of 15 min suggests that this method can identify ESBL-producing strains much more rapidly than conventional methods.

Remarkable Effects of External Magnetic Field on Circularly Polarized Luminescence of EuIII (hfa)3 with Phosphine Chirality.

Herein, magnetic circularly polarized luminescence (CPL) (MCPL) spectroscopy was conducted to analyze an EuIII (hfa)3 complex with three chiral PIII -ligands. Resultantly, (R)-chirality luminophores with S-up orientation and (S)-chirality luminophores with N-up orientation were observed to possess symmetrical mirror image spectra, i. e., they were enantiomers. Similarly, the (R)-chirality luminophores with N-up orientation and the (S)-chirality luminophores with S-up orientation were also enantiomers. Contrarily, (R)-S-up and (S)-S-up were diastereomers and did not possess a mirror-image relationship. Likewise, (R)-N-up and (S)-N-up were diastereomers. The J-dependency of gMCPL and gCPL datasets suggested that the N-up/S-up external magnetic field, with the aid of chiral PIII -ligands, increased the gMCPL values by two- to sixteen-fold and modulated the gMCPL signs at J=1-4. Additionally, the origins of the nonideal mirror-symmetric CPL and MCPL spectral characteristics of EuIII (hfa)3 with three chiral PIII -ligands were discussed in terms of parity (space-inversion, P)-symmetry, time-reversal (T)-symmetry, and PT-symmetry laws.

Excimer-origin CPL vs. monomer-origin magnetic CPL in photo-excited chiral binaphthyl-ester-pyrenes: critical role of ester direction.

Two chiral binaphthyl (BNp) derivatives bearing oppositely oriented ester linkers to two pyrene (Py) moieties [(R)/(S)-1 and (R)/(S)-2] enabled Py-origin circularly polarized luminescence (CPL), magnetic CPL (MCPL), and circular dichroism (CD). (R)-1 that exhibited (-)-sign CD showed (+)-sign Py-excimer CPL but did not exhibit MCPL. Conversely, (R)-2, with (-)-sign CD, did not show excimer-origin CPL, but exhibited clear Py-monomer MCPL.

Analysis of Clinical Isolates of Extended-Spectrum ß-Lactamase-Producing Bacteria with Primer and Probe Sets Developed to Detect blaCTX-M, blaTEM, and blaSHV Using a Fully Automated Gene Detection System.

In this study, we evaluated extended-spectrum ß-lactamase (ESBL)-producing bacteria with the newly developed primer and probe sets to detect blaCTX-M, blaTEM, and blaSHV using BD MAXTM, a fully automated multiplex polymerase chain reaction assay system. In 36 isolates confirmed by whole-genome sequencing to have blaCTX-M, blaTEM, or blaSHV, the developed primer and probe sets accurately detected each gene without being influenced by the presence of other ß-lactamase genes. In nine control strains that do not harbor either blaCTX-M, blaTEM, or blaSHV no cross-reaction was observed. In 191 strains phenotypically determined to be ESBL-producers by conventional antimicrobial susceptibility tests, 189 strains were blaCTX-M-, blaTEM-, or blaSHV-positive as assessed by BD MAXTM using the developed primer and probe sets, and two strains were negative for these genes. Whole-genome sequencing revealed that these two strains were phenotypically false-positive ESBL-producers. The accuracy of the primer and probe sets seems to be satisfactory, and they may be applicable to detect CTX-M-type ESBL-producing bacteria.

Solid-state AIEnh-circularly polarised luminescence of chiral perylene diimide fluorophores.

Solid-state organic fluorescent materials are important for the development of electroluminescent sensing devices. Herein, we report that N,N'-bis((R)-1-phenylethyl)perylene-3,4,9,10-tetracarboxylic diimide [(R,R)-BPP] and its antipode [(S,S)-BPP], which contain extended π-electrons through planar perylenes, emit solid-state aggregation-induced-enhanced (AIEnh) circularly polarised luminescence (CPL) in inorganic (KBr) pellets and organic-polymer-film (PMMA- and myo-IPU-film) states; this CPL is difficult to observe in solution. These chiral perylene fluorophores emit AIEnh-CPL with high dissymmetry factors (g CPL) (up to 2.4 × 10-3) and high quantum yields (Φ F, up to 0.43) in the three solid matrices.

Characterization of Mucoid and Non-Mucoid Streptococcus pneumoniae Isolated From Outpatients.

BACKGROUND: Streptococcus pneumoniae causes pneumonia, sepsis, and meningitis. This study aimed to investigate the clinical characteristics of mucoid and non-mucoid isolates of S. pneumoniae, and to explore the relationship between the isolate phenotypes and their antibiotic susceptibility. METHODS: Clinical isolates from 3,453 non-repetitive S. pneumoniae (189 mucoid and 3,264 non-mucoid) infections obtained between January 2008 and December 2012 from outpatients at the Kimitsu-Central Hospital were evaluated. RESULTS: Compared to the non-mucoid isolates, the mucoid phenotypes were more susceptible to certain antibiotics such as erythromycin, clarithromycin, and tetracycline as opposed to clindamycin, chloramphenicol, and rifampicin. The mucoid phenotype was isolated more frequently from schoolchildren, adults, and elderly adults in a variety of clinical sites, including otorrhea, genitalia, pus, and eye discharge than the non-mucoid phenotype. This suggested that mucoid isolates are more likely to be involved than non-mucoid isolates in various local infections. Systemic infection, which indicates invasiveness, was not associated with the mucoid or non-mucoid phenotype. CONCLUSIONS: The results of this study suggest that mucoid isolates tend to have higher susceptibility than non-mucoid isolates to antibiotics. To the best of our knowledge, mucoid and non-mucoid S. pneumoniae isolates considerably differ in terms of clinical isolation site and age-specific prevalence.