[A Case of Severe Legionella longbeachae Pneumonia and Usefulness of LAMP Assay].

Urinary antigen test is frequently used as a routine laboratory test for early diagnosis of Legionella infection, which is especially suitable for ordinary Legionella pneumophila serogroup 1, but not for other types of Legionella. We report a case of severe pneumonia caused by Legionella longbeachae, where a method of loop-mediated isothermal amplification (LAMP) assay contributed an important role for the early detection. This case involved an 83-year-old man who developed fever, dyspnea, and productive cough. Since the medication of prescribed ceftriaxone had not been effective, he visited the emergency room of our hospital, where an X-ray revealed a severe pneumonia harboring a consolidation with air bronchogram in his right lower lung. His sputum and urine were subjected to the routine bacterial culture or the urinary antigen test for Legionella, which initially brought negative results. However, a positive result of LAMP assay enabled early diagnosis of Legionella pneumonia. Later, the bacterial cultures of sputum made some progress and 16S rRNA sequencing provided a proof of L. longbeachae. This LAMP assay may bring a benefit for the patients with Legionella pneumonia by enabling early detection of not only specific L. pneumophila serogroup 1, but also of the other Legionella species.

[One-year surveillance study of antimicrobial resistance in major bacteria except for MRSA isolated in eight medical facilities in Kumamoto prefecture].

To investigate the current status of drug-resistant bacteria (except MRSA) in Kumamoto prefecture, a study was conducted to determine the isolation numbers and ratios of extended-spectrum beta-lactamase (ESBL)-Escherichia coli, ESBL-Klebsiella species, ESBL-Proteus mirabilis, two-drug resistant Pseudomonas aeruginosa, (resistant to two drugs either carbapenems, quinolones and aminoglycosides) multi-drug-resistant Pseudomonas aeruginosa, multi-drug-resistant Acinetobacter baumannii, and vancomyacin-resistant Enterococcii in eight general hospitals from May in 2009 to April in 2010. ESBL-E. coli was mostly isolated, and two-drug resistant P. aeruginosa came second. The isolation ratio of overall drug-resistant bacteria did not increase, while the isolation ratio of two-drugs resistant P. aeruginosa declined, suggesting that infection control was successfully conducted in these hospitals. However, the isolation numbers of ESBL-Klebsiella spp. and two-drug resistant P. aeruginosa were variable in each hospital. Furthermore, drug-resistant bacteria were occasionally spread into another medical facilities by patients transferred from these hospitals, indicating that sharing information on drug-resistant bacteria between medical facilities is required.

Bhc-cNMPs as either water-soluble or membrane-permeant photoreleasable cyclic nucleotides for both one- and two-photon excitation.

Cyclic nucleoside monophosphates (cNMPs) play key roles in many cellular regulatory processes, such as growth, differentiation, motility, and gene expression. Caged derivatives that can be activated by irradiation could be powerful tools for studying such diverse functions of intracellular second messengers, since the spatiotemporal dynamics of these molecules can be controlled by irradiation with appropriately focused light. Here we report the synthesis, photochemistry, and biological testing of 6-bromo-7-hydroxycoumarin-4-ylmethyl esters of cNMP (Bhc-cNMP) and their acetyl derivatives (Bhc-cNMP/Ac) as new caged second messengers. Irradiation of Bhc-cNMPs quantitatively produced the parent cNMPs with one-photon uncaging efficiencies (Phiepsilon) of up to one order of magnitude better than those of 2-nitrophenethyl (NPE) cNMPs. In addition, two-photon induced photochemical release of cNMP from Bhc-cNMPs (7 and 8) can be observed with the two-photon uncaging action cross-sections (delta(u)) of up to 2.28 GM (1 GM=10(-50) cm(4) s photon(-1)), which is the largest value among those of the reported Bhc-caged compounds. The wavelength dependence of the delta(u) values of 7 revealed that the peak wavelength was twice that of the one-photon absorption maximum. Bhc-cNMPs showed practically useful water solubility (nearly 500 microM), whereas 7-acetylated derivatives (Bhc-cNMPs/Ac) were expected to have a certain membrane permeability. Their advantages were demonstrated in two types of biological systems: the opening of cAMP-mediated transduction channels in newt olfactory receptor cells and cAMP-mediated motility responses in epidermal melanophores in scales from medaka fish. Both examples showed that Bhc and Bhc/Ac caged compounds have great potential for use in many cell biological applications.