Asymmetric Cationic Polymerization of Benzofuran through a Reversible Chain-Transfer Mechanism: Optically Active Polybenzofuran with Controlled Molecular Weights.

Benzofuran (BzF) is a prochiral, 1,2-disubstituted, unsymmetric cyclic olefin that can afford optically active polymers by asymmetric polymerization, unlike common acyclic vinyl monomers. Although asymmetric cationic polymerization of BzF was reported by Natta et al. in the 1960s, the polymer structure has not been clarified, and there are no reports on molecular weight control. Herein, we report dual control of the optical activity and molecular weight of poly(BzF) using thioether-based reversible chain-transfer agents for asymmetric cationic polymerization with ß-amino acid derivatives as chiral additives and aluminum chloride as a catalyst. This asymmetric moderately living cationic polymerization leads to an increase in molecular weight and specific optical rotation with monomer conversion. In addition, asymmetric block polymers consisting of opposite absolute configurational segments were synthesized using both enantiomers sequentially as chiral additives. Finally, a comprehensive analysis of the polymerization products and the model reaction revealed that the optical activity of poly(BzF) originates from the threo-diisotactic structure, which occurs by regio-, trans-, and enantioselective propagation.

A change in the timing of the Bacillus Calmette-Guérin vaccination in 2013 was associated with an increase in the incidence rate of infants with latent tuberculosis infection.

BACKGROUND: A change in the timing of the Bacillus Calmette-Guérin (BCG) vaccination among infants in Japan appears to be associated with an increase in the incidence rate of latent tuberculosis infection (LTBI). METHODS: Data on both active and latent tuberculosis (TB) infections from 2007 to 2019, which were reported by the Japan Anti-Tuberculosis Association, were statistically analyzed by comparing TB incidence rates in 2007-2012 and 2013-2019. RESULTS: Although the incidence rate for active TB disease did not statistically increase nor decrease in the infant age group for either sex (and in fact decreased for some of the other age groups), the incidence rates of LTBI for both sexes were increased in the infant age group, while the incidence rates decreased in the other age groups. Between 2007 and 2012, the incidence rate of LTBI in females was statistically greater than those of males in the 1-4-year-old age group. From 2013 to 2019, the incidence rates of females were greater than those of males in both the infant and 1-4-year-old age groups, suggesting a growing preponderance of infections among female children in the youngest age groups. CONCLUSION: It may be that the change of BCG vaccination timing in Japan which took place in 2013 affected the infant incidence rate of LTBI, with a more prominent effect on females than males. In order to control TB infection, the ramifications of a change in vaccination timing therefore need careful exploration, as one such change appears associated with increased numbers of infants with LTBI, with disproportionate effects on females.

Characteristics and etiology of infection in febrile infants aged ≤90 days.

BACKGROUND: Infants ≤90 days old can exhibit non-specific signs of infection, even in cases of serious bacterial infection (SBI). METHODS: This prospective study included infants aged ≤90 days hospitalized for fever from June 2017 to August 2019. Nasopharyngeal swabs were tested using multiplex real-time polymerase chain reaction (PCR) tests and 16S ribosomal RNA analysis of whole blood to determine causative microorganisms. Data pertaining to inflammatory markers, maximum body temperature (BT), and respiratory symptoms of infants and their cohabiting families were collected at admission. RESULTS: A total of 110 infants were enrolled (age range, 9-90 days), 17 (15.5%) of whom presented with SBIs. White blood cell (WBC) count and absolute neutrophil count (ANC) were significantly higher in patients with SBIs than in those without, although maximum BT did not significantly differ between the SBI and non-SBI groups (n = 93). One or more viruses were detected in 82 infants (74.5%). Viruses were detected more frequently in infants with respiratory symptoms than in those without respiratory symptoms (P = 0.038), and patients with SBIs experienced significantly less respiratory symptoms than those without SBIs (P = 0.049). Moreover, viruses were more often detected in infants from cohabiting families with respiratory symptoms than in those whose family members did not exhibit respiratory symptoms (P = 0.0018). CONCLUSION: White blood cell count, and ANC were significantly higher, and respiratory symptoms were less in infants ≤90 days old with SBIs than in those without SBIs. Microorganisms from nasopharyngeal by multiplex real-time PCR swabs could not be judged as SBI or non-SBI.

Stepwise neuronal network pattern formation in agarose gel during cultivation using non-destructive microneedle photothermal microfabrication.

Conventional neuronal network pattern formation techniques cannot control the arrangement of axons and dendrites because network structures must be fixed before neurite differentiation. To overcome this limitation, we developed a non-destructive stepwise microfabrication technique that can be used to alter microchannels within agarose to guide neurites during elongation. Micropatterns were formed in thin agarose layer coating of a cultivation dish using the tip of a 0.7 [Formula: see text]-diameter platinum-coated glass microneedle heated by a focused 1064-nm wavelength infrared laser, which has no absorbance of water. As the size of the heat source was 0.7 [Formula: see text], which is smaller than the laser wavelength, the temperature fell to 45 [Formula: see text] within a distance of 7.0 [Formula: see text] from the edge of the etched agarose microchannel. We exploited the fast temperature decay property to guide cell-to-cell connection during neuronal network cultivation. The first neurite of a hippocampal cell from a microchamber was guided to a microchannel leading to the target neuron with stepwise etching of the micrometer resolution microchannel in the agarose layer, and the elongated neurites were not damaged by the heat of etching. The results indicate the potential of this new technique for fully direction-controlled on-chip neuronal network studies.

Emergent synchronous beating behavior in spontaneous beating cardiomyocyte clusters.

We investigated the dominant rule determining synchronization of beating intervals of cardiomyocytes after the clustering of mouse primary and human embryonic-stem-cell (hES)-derived cardiomyocytes. Cardiomyocyte clusters were formed in concave agarose cultivation chambers and their beating intervals were compared with those of dispersed isolated single cells. Distribution analysis revealed that the clusters' synchronized interbeat intervals (IBIs) were longer than the majority of those of isolated single cells, which is against the conventional faster firing regulation or "overdrive suppression." IBI distribution of the isolated individual cardiomyocytes acquired from the beating clusters also confirmed that the clusters' IBI was longer than those of the majority of constituent cardiomyocytes. In the complementary experiment in which cell clusters were connected together and then separated again, two cardiomyocyte clusters having different IBIs were attached and synchronized to the longer IBIs than those of the two clusters' original IBIs, and recovered to shorter IBIs after their separation. This is not only against overdrive suppression but also mathematical synchronization models, such as the Kuramoto model, in which synchronized beating becomes intermediate between the two clusters' IBIs. These results suggest that emergent slower synchronous beating occurred in homogeneous cardiomyocyte clusters as a community effect of spontaneously beating cells.

Epidemiology of urinary tract infections in children: Causative bacteria and antimicrobial therapy.

BACKGROUND: Urinary tract infections (UTIs) are the most common bacterial infections in children. This study aimed to review characteristics of causative bacteria and the effectiveness of antimicrobial therapy in children with febrile UTIs. METHODS: Clinical records of 108 patients (130 episodes) with febrile UTIs admitted to the Kawasaki Medical School Hospital between July 2009 and October 2016 were retrospectively reviewed. The characteristics of the causative bacteria, antibacterial therapy, and therapeutic effect were verified. RESULTS: Patients were aged between 0 and 183 months (median age: 3 months). Seventy-three (67.6%) were males. Sixty-three episodes (48.5%) were diagnosed with complicated UTIs. Forty-seven episodes (36.2%) were observed in patients aged <3 months; 15 of them had complicated UTIs. Escherichia coli (E. coli) was the most common pathogen, followed by Enterococcus faecalis (E. faecalis). Blood cultures were positive in three episodes. Among the 130 episodes, 62 (47.7%) were treated with a combination of ampicillin and third-generation cephalosporins, followed by third-generation cephalosporins (31 episodes, 23.8%) and sulbactam sodium / ampicillin sodium (15 episodes, 11.5%). In case of patients with uncomplicated/complicated UTIs and patients aged <3 and ≥3 months, the most common pathogen was E. coli, followed by E. faecalis. There was no difference in therapeutic effects between "combination ampicillin and third-generation cephalosporins" and "third-generation cephalosporin monotherapy" administered for the treatment of UTIs caused by E. coli. CONCLUSIONS: Escherichia coli is the most common pathogen among pediatric UTIs. For antibacterial therapy, third-generation cephalosporin monotherapy is effective and may not require combination therapy with ampicillin.

Recent acute reduction in macrolide-resistant Mycoplasma pneumoniae infections among Japanese children.

INTRODUCTION: Mycoplasma pneumoniae contributes to numerous pneumonia cases among children and young adults. Therefore, this study aimed to investigate the prevalence of M. pneumoniae infections among Japanese children, occurring since 2008. METHODS: Nasopharyngeal swab specimens were obtained from all cases, following which real-time PCR was performed to identify M. pneumoniae. Further, the p1 genotypes of isolates were determined using the PCR restriction fragment length polymorphism typing method. RESULTS: The annual rate of macrolide-resistant M. pneumoniae (MRMP) infections peaked at 81.8% in 2012 and decreased annually until 2015. Although the infection rate increased to 65.3% in 2016, it decreased again to 14.3% in 2018. Although >90% of isolates harbored the type 1 genotype until 2012, this rate decreased, and approximately 80% harbored p1 genotypes other than type 1 in 2018. Furthermore, the occurrence rate of MRMP among the type 1 isolates was very high (82.4%), whereas that among p1 genotypes other than type 1 was very low (6.5%). CONCLUSIONS: MRMP occurrence potentially decreased owing to changes in not only antibiotic usage but also in the distribution of p1 genotype among isolates.

Geometric Understanding of Local Fluctuation Distribution of Conduction Time in Lined-Up Cardiomyocyte Network in Agarose-Microfabrication Multi-Electrode Measurement Assay.

We examined characteristics of the propagation of conduction in width-controlled cardiomyocyte cell networks for understanding the contribution of the geometrical arrangement of cardiomyocytes for their local fluctuation distribution. We tracked a series of extracellular field potentials of linearly lined-up human embryonic stem (ES) cell-derived cardiomyocytes and mouse primary cardiomyocytes with 100 kHz sampling intervals of multi-electrodes signal acquisitions and an agarose microfabrication technology to localize the cardiomyocyte geometries in the lined-up cell networks with 100-300 µm wide agarose microstructures. Conduction time between two neighbor microelectrodes (300 µm) showed Gaussian distribution. However, the distributions maintained their form regardless of its propagation distances up to 1.5 mm, meaning propagation diffusion did not occur. In contrast, when Quinidine was applied, the propagation time distributions were increased as the faster firing regulation simulation predicted. The results indicate the "faster firing regulation" is not sufficient to explain the conservation of the propagation time distribution in cardiomyocyte networks but should be expanded with a kind of community effect of cell networks, such as the lower fluctuation regulation.

Yersinia pseudotuberculosis Infection Complicated with Bacteremia in a 10-Month-Old Boy.

Yersinia pseudotuberculosis (Y. pseudotuberculosis) infection complicated with bacteremia rarely occurs. Y. pseudotuberculosis infection is also known to produce various symptoms similar to Kawasaki disease (KD) due to the production of Y. pseudotuberculosis-derived mitogen (YPM), an exotoxin with superantigen activity. Moreover, it causes terminal ileitis and is responsible for appendix swelling. Here, we report a case of Y. pseudotuberculosis infection in a 10-month-old boy who was brought to our hospital due to fever, watery stool, and poor vitality. Abdominal echocardiography revealed wall thickening of the entire colon and appendix swelling; therefore, he was admitted and treated with antibiotics for bacterial enteritis or appendicitis. After the antibiotic administration, facial skin rashes and hand and foot edema developed. However, he had 5/6 major symptoms of KD and was diagnosed with Y. pseudotuberculosis infection because of its presence in the blood and stool cultures. Thereafter, antibacterial therapy improved his symptoms and increased the inflammatory response. After his hospital discharge, the skin on his fingers showed desquamation like that of KD. Y. pseudotuberculosis infection should be considered as a differential disease in KD, terminal ileitis, and appendicitis. Furthermore, its infection route and culture methods should also be carefully considered.

Low prevalence of Chlamydia pneumoniae infections during the Mycoplasma pneumoniae epidemic season: Results of nationwide surveillance in Japan.

OBJECTIVE: Chlamydia pneumoniae and Mycoplasma pneumoniae are both common causes of atypical pneumonia. We conducted an annual national survey of Japanese children to screen them for C. pneumoniae infections during the M. pneumoniae epidemic season. METHODS: Nasopharyngeal swab specimens were collected from children aged 0-15 years with suspected acute lower respiratory tract infection due to atypical pathogens, at 85 medical facilities in Japan from June 2008 to March 2018. Specimens were tested for infection using real-time polymerase chain reaction assays. RESULTS: Of 5002 specimens tested, 1822 (36.5%) were positive for M. pneumoniae alone, 42 (0.8%) were positive for C. pneumoniae alone, and 20 (0.4%) were positive for both organisms. In children with C. pneumoniae infection, the median C. pneumoniae DNA copy number was higher in those with single infections than in those with M. pneumoniae coinfection (p = 0.08); however it did not differ significantly according to whether the children had received antibiotics prior to sample collection (p = 0.34). CONCLUSIONS: The prevalence of C. pneumoniae infection was substantially lower than that of M. pneumoniae infection during the study period. The change in prevalence of C. pneumoniae was not influenced by that of M. pneumoniae. Children with single C. pneumoniae infection are likely to have had C. pneumoniae infection, while those with coinfection are likely to have been C. pneumoniae carriers.

Molecular epidemiology, antimicrobial susceptibility, and characterization of fluoroquinolone non-susceptible Streptococcus pyogenes in Japan.

Streptococcus pyogenes (Group A streptococci: GAS) are known to cause a wide variety of human illnesses, some of which can be life-threatening. Usually, penicillin is the first-choice agent for the treatment of GAS infections. For patients with penicillin or beta-lactam antibiotics allergies, macrolide drugs are recommended as an alternative therapy. However, an increased prevalence of macrolide-resistant GAS (MRGAS) has been reported in many countries. Furthermore, fluoroquinolone non-susceptible GAS has been reported. The present study was focused on determining the features of fluoroquinolone non-susceptible strains collected from children with pharyngotonsillitis in the southwestern areas of Japan. To reveal the characteristics of fluoroquinolone non-susceptible GAS, we investigated the MIC, T-serotype, emm typing, and PFGE of 298 GAS strains isolated in the Fukuoka southwest area of Japan between 2011 and 2013. We determined that fluoroquinolone non-susceptibility shows a MIC to tosufloxacin of ≧1 µg/ml. We identified 33 (11.1%) fluoroquinolone non-susceptible GAS strains. In these strains, 6 T-serotypes and 9 emm/MLST patterns were detected. The predominant combinations were emm6/ST382 (14 strains, 42.4%) and emm89/ST101 (5 strains, 15.2%). PFGE classified 10 pulsotypes, and each was quite different. These results showed that fluoroquinolone non-susceptible GAS strains have a variety of origins. The usage of fluoroquinolone drugs could have a negative effect on the antimicrobial drug sensitivity of GAS in Japan. Considering such a situation, continuous monitoring of quinolone non-susceptible GAS is necessary.

Comparing Antimicrobial Susceptibilities among Mycoplasma pneumoniae Isolates from Pediatric Patients in Japan between Two Recent Epidemic Periods.

We compared the antimicrobial susceptibility of Mycoplasma pneumoniae isolates from pediatric patients in Japan in 2011-2012 and 2015-2016, when epidemics occurred. The antimicrobial activity of macrolides and tetracyclines against M. pneumoniae infection tended to be restored in 2015-2016. There was no change in the antimicrobial activity of quinolones against M. pneumoniae infection.

LAMP-based assay can rectify the diagnosis of Yersinia pseudotuberculosis infections otherwise missed by serology.

BACKGROUND: Despite being a well-known but seldom encountered zoonotic pathogen, diagnosis of Yersinia pseudotuberculosis is not necessarily easy. Infected patients occasionally present with various symptoms resembling Kawasaki disease; thus discriminating the two in the acute phase is challenging. In addition to bacterial culture and serology, novel detection methods based on loop-mediated isothermal amplification (LAMP) are reported in the literature. However, the clinical utility of LAMP-based methods in comparison with the other methods is scarcely documented in the literature. AIM: To clarify the clinical utility of a LAMP-based method in the diagnosis of Yersinia pseudotuberculosis infection. METHODOLOGY: Inpatients admitted due to suspected Yersinia pseudotuberculosis infection during April 2008 through March 2015 were enrolled. Results of the LAMP-based method as well as culture and serology were collected and compared. RESULTS: Among 16 eligible cases, serology proved positive in 13 (81.3 %) cases, LAMP in eight (50 %) cases, and bacterial culture in four (25 %) cases. No significant difference among the three methods could be proved statistically. Although serology was the most sensitive method, it is known to miss cases such as young patients, whereas LAMP could complement all three cases missed by serology. Furthermore, LAMP can return the test result within a few hours from specimen receipt, whereas serology and bacterial culture requires days to weeks of time. CONCLUSION: Although second to serology in sensitivity, the LAMP-based method proved its utility in making rapid diagnosis, and serving a complementary role to serology.

Macrolide-Resistant Mycoplasma pneumoniae Infection, Japan, 2008-2015.

We evaluated isolates obtained from children with Mycoplasma pneumoniae infection throughout Japan during 2008-2015. The highest prevalence of macrolide-resistant M. pneumoniae was 81.6% in 2012, followed by 59.3% in 2014 and 43.6% in 2015. The prevalence of macrolide-resistant M. pneumoniae among children in Japan has decreased.

Molecular epidemiology, antimicrobial susceptibility, and characterization of macrolide-resistant Streptococcus pyogenes in Japan.

Here we report the molecular epidemiology of macrolide-resistant Streptococcus pyogenes (group A streptococci, GAS) isolated from children with pharyngotonsillitis between 2011 and 2013 in Japan. In 299 isolates, 124 (41.5%) isolates were macrolide-resistant. We characterized the isolates by emm typing, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Of 299 isolates, 124 (41.5%) were macrolide-resistant isolates, 76 (61.3%) possessed mefA and 46 (37.1%) possessed ermB. All 76 isolates with mefA possessed msrD. There were no isolates possessed ermTR in this study. Eight emm/MLST types were observed. The predominant type was emm1/ST28 (57 isolates, 46.0%), which possessed the mefA/msrD complex, presenting as the M phenotype. The second most predominant type was emm12/ST467, which possessed ermB, presenting as the cMLSB phenotype. Of the cMLSB phenotype isolates, types emm28/ST52 and emm12/ST36 had multiple genetic backgrounds. We found high proportions of macrolide-resistant GAS in the southwestern areas of Japan.

An HIV-infected Pregnant Woman Treated with the Long-term Administration of Antiretroviral Therapy Including Raltegravir.

A 27-year-old HIV-infected pregnant Japanese woman was admitted to our hospital at gestational week 14. The patient's HIV viral load was 71,000 copies/mL, and her CD4 cell count was 147 cells/mm(3). Zidovudine, lamivudine, and lopinavir/ritonavir were administered at gestational week 18. Because the viral load increased to 222,000 copies/mL at the initiation of antiretroviral therapy, we added raltegravir. The decrease in the viral load was satisfactory, and a caesarean delivery was performed. Although the plasma concentration of raltegravir in the neonate was significantly high (2,482 ng/mL), no adverse event was confirmed. There was no evidence of the mother-to-child transmission of HIV.

[Antibiotic therapy against acute tonsillopharyngitis in children due to group A beta-hemolytic streptococci: comparison of clinical efficacy, the bactericidal effects, and effects on oral flora between cefditoren pivoxil for 5 days and amoxicillin for 10 days].

We compared the clinical efficacy, the bactericidal effects, effect on the oral microbial flora, and adverse reactions between cefditoren pivoxil (CDTR-PI) for 5 days and amoxicillin (AMPC) for 10 days in children with acute group A beta-hemolytic streptococci (GAS) tonsillopharyngitis, and simultaneously examined the emm genotype and drug susceptibility of the isolated GAS. The results showed that the clinical efficacy was 100% for CDTR-PI and 97.9% for AMPC, with no difference between the two groups, and the bacterial elimination rate was 100% in both groups. No serious adverse event was noted in either group. On the other hand, concerning changes in the oral microbial flora between before and after treatment, the amount of bacteria showed no change in the CDTR-PI group (p = 0.5761) but clearly decreased in the AMPC group (p = 0.0049). This indicates that CDTR-PI does not disturb the oral microbial flora compared with AMPC. Also, the emm types determined in the 112 GAS strains isolated in this study were similar to those that have recently been isolated frequently in Japan. Concerning the drug resistance, none of the isolates showed resistance to beta-lactam antibiotics, but 45% of them were resistant to macrolides. The advantages of short-term treatment are considered to include a lower cost, improvement in drug compliance, decrease in the frequency of the occurrence of adverse reactions, decrease in the frequency of the appearance of drug-resistant strains, and alleviation of the psychological burden of patients and their parents. For these reasons, we conclude that CDTR-PI for 5 days is a useful option for the treatment of acute GAS tonsillopharyngitis in children.

Power-law analysis of surface-plasmon-enhanced electromagnetic field dependence of blinking SERS of thiacyanine or thiacarbocyanine adsorbed on single silver nanoaggregates.

Blinking statistics in surface-enhanced Raman scattering (SERS) of thiacyanine or thiacarbocyanine adsorbed on single Ag nanoaggregates were analyzed by a power law. A power law reproduces the probability distributions of both the bright and dark SERS occurrences against their duration times. As the localized surface plasmon resonance (LSPR) wavelength of a single Ag nanoaggregate approached the excitation wavelength or the excitation laser intensity increases, the power-law exponents were close to -1.5, a value derived from a one-dimensional random walk model. When the LSPR wavelength left the excitation wavelength or the excitation laser intensity decreases, the power-law exponents deviated from -1.5. The decrease in the power-law exponents in the bright SERS, which indicates a decrease in the probabilities of the long-lived bright SERS, and the increase in the power-law exponents in the dark SERS coincide with the increasing shallowness and narrowing of a optical trapping potential well due to a surface-plasmon-enhanced electromagnetic field around a junction of the Ag nanoaggregates excited at a wavelength apart from the LSPR wavelength or under the low laser intensity, i.e., the low original electromagnetic field, respectively.

Analysis of excitation laser intensity dependence of blinking SERRS of thiacarbocyanine adsorbed on single silver nanoaggregates by using a power law with an exponential function.

In blinking SERRS, probability distributions of the dark events against their duration were mostly reproduced by a power-law with the same exponential function under various excitation laser intensities, although they were given by a power-law with the different exponential functions at other excitation wavelengths.