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Vonoprazan is a potent inhibitor of gastric acid secretion and may have better response than proton pump inhibitors (PPIs) in the treatment of endoscopic submucosal dissection induced artificial ulcers. However, reported outcomes remain controversial. In this study, we conducted a prospective, randomized comparative trial to evaluate healing effects of vonoprazan and lansoprazole on endoscopic submucosal dissection (ESD)-induced ulcers. We enrolled 216 patients who underwent endoscopic submucosal dissection for early gastric neoplasms. They were randomly divided into vonoprazan (20 mg/day) and lansoprazole (30 mg/day) groups. The primary endpoint was the reduction rate of ulcer and complete healing (scar) ratio of ESD-induced ulcers at 4 and 8 weeks. Finally, 101 patients of the vonoprazan group and 95 patients of the lansoprazole group were included in the analysis. There were no significant differences in the reduction rate between the vonoprazan and lansoprazole groups at either timepoint (4 weeks, 94.0 vs 93.4%; 8 weeks, 99.8 vs 99.9%, respectively). The complete healing ratio at 4 and 8 weeks did not differ significantly between the vonoprazan and lansoprazole groups (4 weeks, 11.9 vs 12.6%; 8 weeks, 87.1 vs 86.3%, respectively). In the anti-H. pylori-antibody negative or positive patients, there were no significant differences in the reduction rate and complete healing ratio between the vonoprazan and lansoprazole groups. Regardless of treatment choice, the overall complete healing ratio at 8 weeks was significantly higher in the anti-H. pylori-antibody negative patients than the positive patients (p = 0.006). The healing effects of vonoprazan on ESD-induced ulcers were comparative to those of lansoprazole.

A 44-year-old man with a history of chronic alcoholic pancreatitis and Crohn's disease presented with abdominal pain. Computed tomography revealed pancreatic calculi in the head of the pancreas and a dilated pancreatic duct. The patient was diagnosed with an acute exacerbation of chronic pancreatitis due to the impact of pancreatic calculi on the main pancreatic duct. During the clinical course, the movement of pancreatic calculi to the major papilla was confirmed, leading to obstructive jaundice. Endoscopic treatment with sphincterotomy of the pancreatic duct was successful. Herein, we report the case of an unusual clinical course involving obstructive jaundice caused by the movement of pancreatic calculi.

Background Enterovirus D68 (EV-D68) causes asthma-like respiratory infection in children. Several EV-D68 outbreaks have been reported worldwide since the largest outbreak occurred in the United States in 2014. We experienced an accumulation of pediatric cases with asthma-like respiratory illness in Niigata, Japan, in 2018. Study design To determine whether EV-D68 was responsible for the case accumulation, this prospective observational study evaluated children hospitalized in 1 of 8 hospitals with asthma-like respiratory illness in Niigata, Japan, during October and November 2018. Diagnoses were made by EV-D68-specific RT-PCR using nasopharyngeal samples. The clade was identified by sequence analyses, and a phylogenetic tree was created. To evaluate seasonal variation, data from pediatric cases with asthma-like respiratory illness in 2018 were retrospectively analyzed. Results In 2018, 114 children were hospitalized with asthma-like respiratory illness in October and November, and 47 nasopharyngeal samples were collected. EV-D68 was detected in 22/47 (47%) patients during the study period. The phylogenetic tree revealed that all strains belonged to the clade B3 branch, which has been detected worldwide every 2 years since 2014. Conclusions EV-D68 was the associated pathogen for asthma-like respiratory illness in children in Japan in 2018. Clade B3, the dominant clade in outbreaks worldwide, was responsible for the outbreak. Detection and detailed virologic analysis of EV-D68 is important as part of worldwide surveillance, as it will aid in understanding the epidemiologic characteristics of EV-D68 infection.

Background/aims There are few prospective studies on cold forceps polypectomy (CFP) using jumbo cup forceps. Therefore, we examined patients with diminutive polyps (5 mm or smaller) treated with CFP using jumbo cup forceps to achieve an adenoma-free colon and also assessed the safety of the procedure and the recurrence rate of missed or residual polyp after CFP by performing follow-up colonoscopy 1 year later. Methods We included patients with up to 5 adenomas removed at initial colonoscopy and analyzed data from a total of 361 patients with 573 adenomas. One-year follow-up colonoscopy was performed in 165 patients, at which 251 lesions were confirmed. Results The one-bite resection rate with CFP was highest for lesions 3 mm or smaller and decreased significantly with increasing lesion size. Post-procedural hemorrhage was observed in 1 of 573 lesions (0.17%). No perforation was noted. The definite recurrence rate was 0.8% (2/251 lesions). The probable recurrence rate, which was defined as recurrence in the same colorectal segment, was 17%. Adenoma-free colon was achieved in 55% of patients at initial resection. Multivariate analysis revealed that achievement of an adenoma-free colon was significantly associated with number of adenomas and years of endoscopic experience. Conclusions CFP using jumbo biopsy forceps was safe and showed a high one-bite resection rate for diminutive lesions of 3 mm or smaller. The low definite recurrence rate confirms the reliability of CFP using jumbo biopsy forceps. Number of adenomas and years of endoscopic experience were key factors in achieving an adenoma-free colon.

To prospectively investigate the efficacy and safety of clip-flap assisted endoscopic submucosal dissection (ESD) for gastric tumors.From May 2015 to October 2016, we enrolled 104 patients with gastric cancer or adenoma scheduled for ESD at Shiga University of Medical Science Hospital. We randomized patients into two subgroups using the minimization method based on location of the tumor (upper, middle or lower third of the stomach), tumor size (20 mm or20 mm) and ulcer status: ESD using an endoclip (the clip-flap group) and ESD without an endoclip (the conventional group). Therapeutic efficacy (procedure time) and safety (complication: Gastrointestinal bleeding and perforation) were assessed.For patients with early-stage gastric tumors, the clip-flap method has no advantage in efficacy or safety compared with the conventional method.

AIM To investigate factors associated with the healing of endoscopic submucosal dissection (ESD)-induced ulcers. METHODS We enrolled 132 patients with gastric tumors scheduled for ESD. Following ESD, patients were treated with daily lansoprazole 30 mg or vonoprazan 20 mg. Ulcer size was endoscopically measured on the day after ESD and at 4 and 8 wk. The gastric mucosa was endoscopically graded according to the Kyoto gastritis scoring system. We assessed the number of patients with and without a 90% reduction in ulcer area at 4 wk post-ESD and scar formation at 8 wk, and looked for risk factors for slower healing. RESULTS The mean size of gastric tumors and post-ESD ulcers was 17.4 ± 12.1 mm and 32.9 ± 13.0 mm. The mean reduction rates in ulcer area were 90.4% ± 0.8% at 4 wk and 99.8% ± 0.1% at 8 wk. The reduction rate was associated with the Kyoto grade of gastric atrophy at 4 wk (A0: 97.9% ± 0.6%, A1: 93.4% ± 4.1%, and A2: 89.7% ± 1.0%, respectively). In multivariate analysis, the factor predicting 90% reduction at 4 wk was gastric atrophy (Odds ratio: 5.678, 95%CI: 1.190-27.085, P = 0.029). CONCLUSION The healing speed of post-ESD ulcers was associated with the degree of gastric mucosal atrophy, and Helicobacter pylori eradication therapy is required to perform at younger age.

Cap polyposis is a rare intestinal disorder. Characteristic endoscopic findings are multiple inflammatory polypoid lesions covered by caps of fibrous purulent exudate. Although a specific treatment has not been established, some studies have suggested that eradication therapy for Helicobacter pylori (H. pylori) is effective. We report a case of a 20-year-old man with cap polyposis presenting with hematochezia. Colonoscopy showed the erythematous polyps with white caps from the sigmoid colon to rectum. Histopathological findings revealed elongated, tortuous, branched crypts lined by hyperplastic epithelium with a mild degree of fibromusculosis in the lamina propria. Although H. pylori eradication was instituted, there was no improvement over six months. We then performed en bloc excision of the polyps by endoscopic submucosal dissection (ESD), which resulted in complete resolution of symptoms. ESD may be a treatment option for cap polyposis refractory to conservative treatments. We review the literature concerning treatment for cap polyposis and clinical outcomes.

Objective The Kyoto gastritis classification categorizes the endoscopic characteristics of Helicobacter pylori (H. pylori) infection-associated gastritis and identifies patterns associated with a high risk of gastric cancer. We investigated its efficacy, comparing scores in patients with H. pylori-associated gastritis and with gastric cancer. Methods A total of 1,200 patients with H. pylori-positive gastritis alone (n=932), early-stage H. pylori-positive gastric cancer (n=189), and successfully treated H. pylori-negative cancer (n=79) were endoscopically graded according to the Kyoto gastritis classification for atrophy, intestinal metaplasia, fold hypertrophy, nodularity, and diffuse redness. Results The prevalence of O-II/O-III-type atrophy according to the Kimura-Takemoto classification in early-stage H. pylori-positive gastric cancer and successfully treated H. pylori-negative cancer groups was 45.1%, which was significantly higher than in subjects with gastritis alone (12.7%, p

Parechovirus-A3 (PeV-A3) and the enteroviruses (EVs) are the most common viral pathogens responsible for sepsis and meningoencephalitis in neonates and young infants; however, differences in the clinical presentations of two infections are not well described.To describe the clinical presentations of PeV-A3- and EVs-related diseases and develop a novel scoring system to differentiate two diseases.This prospective study used real-time PCR and genetic sequencing to evaluate viral etiologies of febrile neonates and infants4 months with suspected sepsis or meningoencephalitis in Niigata area, Japan, in 2014-2016. The clinical manifestations of PeV-A3- and EVs-infected patients were compared, and a novel scoring system was developed after identifying the most distinguishable clinical findings, followed by the external cohort validation.In 210 patients evaluated, we identified 56 PeV-A3-infected (27%) and 43 EVs-infected (20%) patients. The following clinical manifestations were significant in PeV-A3-infected patients, as compared with EVs-infected patients; a higher body temperature (38.9°C vs. 38.5°C, P .01) and heart rate (181/min vs. 168/min, P = .01), cold extremities (72% vs. 34%, P .01) and skin mottling (65% vs. 23%, P .01), lower white blood cell count (5,200/μL vs. 8,900/μL, P .01) and incidence of cerebrospinal fluid (CSF) pleocytosis (2% vs. 63%, P .01). Using some of these significant findings, the scoring system successfully distinguished the diseases (accuracy: 86% and 83% for the derivative and external validation cohorts, respectively).We found significant clinical manifestations in PeV-A3-infected patients compared to EVs-infected patients. The scoring system may be helpful to distinguish two infections, especially at onset of outbreak.

Moraxella catarrhalis is an exclusively human pathogen that is an important cause of otitis media in children and lower respiratory tract infections in adults with chronic obstructive pulmonary disease. A vaccine to prevent M. catarrhalis infections would have an enormous global impact in reducing morbidity resulting from these infections. Substrate binding protein 2 (SBP2) of an ABC transporter system has recently been identified as a promising vaccine candidate antigen on the bacterial surface of M. catarrhalis . In this study, we showed that SBP1, -2, and -3 individually bind different basic amino acids with exquisite specificity. We engineered mutants that each expressed a single SBP from this gene cluster and showed in growth experiments that SBP1, -2, and -3 serve a nutritional function through acquisition of amino acids for the bacterium. SBP2 mediates uptake of arginine, a strict growth requirement of M. catarrhalis . Adherence and invasion assays demonstrated that SBP1 and SBP3 play a role in invasion of human respiratory epithelial cells, consistent with a nutritional role in intracellular survival in the human respiratory tract. This work demonstrates that the SBPs of an ABC transporter system function in the uptake of basic amino acids to support growth of M. catarrhalis . The critical role of SBP2 in arginine uptake may contribute to its potential as a vaccine antigen.

The current School Health and Safety Act in Japan states that children with influenza infection should stay home until day 6(th) after symptoms onset. This was an amendment of a previous version recommending school return on day 3 after defervescence. Here, we investigated the duration of fever and virus shedding after laninamivir treatment in 7 children infected with influenza A(H3N2) virus and 21 children with influenza B virus in relation to the school return timing recommended by the School Health and Safety Act during the 2011-2012 influenza season. Nasal discharge was collected on the first, second, and third hospital visits and virus titers were assessed by virus culture and real-time PCR. Duration of fever after laninamivir treatment was 1 day longer for influenza B than for influenza A(H3N2). Virus detection rates with 50% tissue culture infectious dose and viral RNA were highest at the first visit and gradually decreased at subsequent visits. Virus positivity rates were detectable at the time of defervescence in less than half of the enrolled patients (14.3-42.9%). Virus shedding rates were similarly low (0.0-19.0%) on day 3 or later from defervescence and on day 6 or later from fever onset (school return dates per the old and current School Health and Safety Act) regardless of the influenza type. In conclusion, despite the higher efficacy of laninamivir against A(H3N2) viruses than B viruses, viral shedding is low after return to school for both types, regardless of the version of the School Health and Safety Act.

BACKGROUNDS AND AIMS Constipation is one of the most common gastrointestinal functional disorders. Recently, the gut microbiota has been implicated in the development of constipation. Helicobacter pylori infection is considered to be a possible factor influencing the gut microbiota profile. Here, we investigated the effect of H. pylori eradication therapy on symptoms of chronic constipation. METHODS We recruited 166 H pylori-positive patients who underwent eradication therapy after endoscopy. We evaluated the severity of symptoms of chronic constipation before eradication therapy and 2 months post-therapy using two questionnaires, the Gastrointestinal Symptom Rating Scale (GSRS) and the Izumo scale. In addition, we evaluated association with constipation and H. pylori infection in patients with constipation-related symptoms in not only all patients, but also patients with the constipation-related symptoms in relation to eradication outcome, the severity of constipation-related symptoms, and the severity of endoscopic gastric mucosal atrophy. RESULTS Mean GSRS scores were 5.10 ± 2.67 in all patients and 6.15 ± 2.91 in constipation patients which were significantly lower than that before eradication (5.78 ± 3.27, P

Background Parechovirus A (PeV-A) is an important cause of sepsis and meningoencephalitis in neonates and young infants. Thus, identifying the source of PeV-A is essential for prevention; however, little is known regarding the spread of PeV-A among family members of PeV-A–infected neonates and young infants. Methods In this prospective study, we evaluated stool samples from family members of PeV-A–infected neonates and infants younger than 4 months who presented with sepsis, meningoencephalitis, or both in Niigata, Japan, in 2016. Because of a simultaneous outbreak, enteroviruses (EVs) were also evaluated during this period. Real-time polymerase chain reaction followed by sequence analysis was used for viral diagnosis using serum and/or cerebrospinal fluid samples. Results Among 54 febrile patients, the stool samples of 14 (26%) and 12 (22%) patients tested positive for PeV-A and EV, respectively. Stool samples from 54 family members (38 adults and 16 children) of 12 PeV-A–infected patients were available. The rate of PeV-A positivity in these samples was higher among the children (88% [14 of 16]) than the adults (34% [13 of 38]). Among family members with a PeV-A–positive stool sample, 29% (4 of 14) of the children and 77% (10 of 13) of the adults were asymptomatic. Similarly, among 53 stool samples from family members (31 adults and 22 children) of 11 EV-infected patients, the rate of EV positivity in the stool samples was higher among the children (91% [20 of 22]) than among the adults (42% [13 of 31]). The asymptomatic-patient rates were 45% (9 of 20) among the children and 85% (11 of 13) among the adults in family members with EV-positive stool. Conclusions Similar to EVs, PeV-A was detected frequently in stool samples from family members of PeV-A–infected patients. Among family members with PeV-A–positive stool, adults were more likely than children to be asymptomatic and therefore could be an important source of PeV-A infection.

Background Parechovirus-A3 (PeV-A3) causes severe disease, including sepsis and meningoencephalitis in young infants. The first case of PeV-A3 was reported in Japan in 1999 and, although epidemics have been reported every 2 to 3 years in more than 20 countries, no major epidemic has occurred in Japan since 2014. Methods This prospective study included febrile infants ( Results Of the 277 patients evaluated, 135 (49%) were positive for PeV-A (n = 74, 27%) or EVs (n = 61, 22%). Among PeV-A patients, most had PeV-A3 (n = 69; 93%), followed by PeV-A4 (n = 4; 5%). There was a PeV-A3 epidemic in 2014 (n = 43); however, no cases were reported in 2015. In 2016–2018, small numbers of PeV-A3 cases were reported: 10 in 2016, 7 in 2017, and 9 in 2018. In contrast, EV cases were reported throughout this period: 8 in 2014, 22 in 2015, 10 in 2016, 5 in 2017, and 16 in 2018. When data were analyzed by season, the PeV-A3 detection rate in summer (June-August) was 93% (40/43) in 2014 and 65% (17/26) during 2016–2018, indicating an increase in the number of PeV-A3 cases in seasons other than summer. Phylogenetic analysis showed that PeV-A3 strains during 2016–2018 were part of a cluster of epidemics in 2011 and differed from those in 2014. Conclusion After the major PeV-A3 epidemics in 2014, we observed changes in the PeV-A3 epidemic profile, namely, a small, but constant, number of cases every year in Niigata, Japan. A future study should investigate if this trend has continued in Japan and other countries and identify the causes of this change in epidemic profile. Disclosures All authors: No reported disclosures.

Background Enterovirus D68 (EV-D68) is a re-emerging, high-morbidity pathogen that causes severe respiratory infection and acute flaccid myelitis in children. EV-D68 is phylogenetically divided into 4 major groups—clade A through D—and each clade has subclades defined by genetic variant. Outbreaks of all strains have been reported in more than 10 countries. However, no study has compared the viral genomic characteristics of EV-D68 in contemporaneous outbreaks in different countries. Methods An outbreak of EV-D68 in children younger than 15 years occurred in Niigata, Japan, from October through November 2018. The patients were admitted to hospital with respiratory distress and wheezing episodes. RNA extracted from nasopharyngeal samples was tested by EV-D68-specific PCR. Clade was determined by semi-nested PCR analysis of the VP1 region, and the phylogenetic tree of the VP1 sequence was constructed. To clarify viral genomic characteristics, we compared the clade to those of outbreaks occurring during 2014–2018 in other countries. Results EV-D68 testing of 47 patients yielded positive results for 22 (47%) (median age 4.6 years; IQR, 2.9–6.7), and 15 (69%) had a past medical history of allergic disease. No patient developed acute flaccid myelitis. The VP1 sequences from all isolates belonged to clade B3, the same clade detected during the 2015 outbreak in Japan (figure). Interestingly, EV-D68 outbreaks were reported in France and Italy (clades D1/B3 and D1, respectively) during this period. Although the EV-D68 clades responsible for outbreaks in 2014 differed by region, clade B3 is now the dominant clade and has caused concurrent EV-D68 outbreaks in children since 2015 (table). Conclusion In 2018, an EV-D68 clade B3 outbreak occurred among children in Niigata, Japan. This clade is now dominant and periodically circulates around the world. Because EV-D68 can cause simultaneous outbreaks worldwide and is associated with high morbidity, detailed real-time epidemiological surveillance of EV-D68 is warranted. Disclosures All authors: No reported disclosures.

Moraxella catarrhalis is a common respiratory tract pathogen that causes otitis media in children and infections in adults with chronic obstructive pulmonary disease. Since the introduction of the pneumococcal conjugate vaccines with/without protein D of nontypeable Haemophilus influenzae , M. catarrhalis has become a high-priority pathogen in otitis media. For the development of antibacterial vaccines and therapies, substrate binding proteins of ATP-binding cassette transporters are important targets. In this study, we identified and characterized a substrate binding protein, SBP2, of M. catarrhalis . Among 30 clinical isolates tested, the sbp2 gene sequence was highly conserved. In 2 different analyses (whole-cell enzyme-linked immunosorbent assay and flow cytometry), polyclonal antibodies raised to recombinant SBP2 demonstrated that SBP2 expresses epitopes on the bacterial surface of the wild type but not the sbp2 mutant. Mice immunized with recombinant SBP2 showed significantly enhanced clearance of M. catarrhalis from the lung compared to that in the control group at both 25-μg and 50-μg doses ( P < 0.001). We conclude that SBP2 is a novel, attractive candidate as a vaccine antigen against M. catarrhalis .

Antibiotic resistance in Streptococcus pneumoniae is a major concern worldwide. However, it is unclear whether resistance is associated with only a few highly prevalent clones or numerous and diverse clones. We monitored 349 healthy children and obtained nasopharyngeal cultures at five time points coinciding with health check-ups (4, 7, 10, 18 and 36 months) between 2008 and 2012. A total of 497 S. pneumoniae isolates from 257 healthy children were characterized using capsular serotyping, multilocus sequence typing and antibiotic resistance genotyping (ermB, mefA/E and pbp mutations). Among these isolates, 25 serotypes and 66 sequence types (STs) were found, including 24 new STs with 11 new alleles. Although resistance was present in a variety of ST clones, most of the clones (57/66, 86.4 %) had one specific resistant or susceptible genotype. Of 233 phenotypically penicillin-non-susceptible isolates, 196 (84.1 %) belonged to only six clones, comprising ST906B, ST23619F, ST24223F, ST37876A, ST143723F and ST33823A and their variants. We concluded that drug-resistant S. pneumoniae is associated with a limited number of highly prevalent clones that are capable of adapting to the community setting. Furthermore, we analysed the capsular gene evolution in serogroup 6. The strain ST29246D was probably the result of recombination of a 3563 bp fragment of the capsule locus acquired by an ST29246C strain from an ST906B or ST29246B strain. Compared with previous studies, our results showed a different recombination site (wciN and wzx) and a different cps profile (8-7-11), indicating that serogroup 6 strains have multiple sites for cps recombination as a mechanism of vaccine escape.

The first step in a bacterial disease is the establishment of nasopharyngeal carriage.We conducted a birth cohort study to identify factors associated with colonization in healthy children and evaluate the serotype distributions and resistances of Streptococcus pneumoniae/Haemophilus influenzae. Nasopharyngeal cultures were obtained from 349 subjects at 5 time points coinciding with health checkups (4, 7, 10, 18 and 36 months).A total of 551 S. pneumoniae (penicillin resistance rate: 46.3%) and 301 H. influenzae (ampicillin resistance rate: 44.5%) isolates were obtained from 1654 samples. In this study, 47.5% and 60.9% of S. pneumoniae isolates were included in the serotypes of 7- and 13-valent pneumococcal conjugate vaccines, respectively. Analyzing by Cox proportional hazards models, cohabiting older sibling(s) attending day-care (hazard ratios: 2.064-3.518, P0.001) and an early start of day-care attendance by the subjects themselves (2.259-2.439, P0.001) were associated with a higher risk of early colonization regardless of their susceptibility. Recent exposure to antimicrobials was also significantly associated with increased risk of colonization (odds ratios: 2.032-2.999, P0.001) but not with resistance rates. This data indicated that introduction of appropriate antimicrobial usage in areas of overuse of antimicrobials could contribute to lower colonization of S. pneumoniae/H. influenzae, resulting in a decrease in the absolute number of resistant isolates.Strategies to control transmission at day-care centers or from older sibling(s) as well as appropriate use of antimicrobials are essential for reducing colonization and the absolute number of resistant isolates.

Persons with untreated atrophic thyroiditis develop a permanent height deficit. Adequate thyroxine replacement therapy can improve growth and the probability of attaining normal adult height. However, height deficit is related to duration of thyroxine deficiency, regardless of the adequacy of thyroxine replacement therapy after diagnosis.1 Several studies reported that use of adjunctive therapy with growth hormone (GH) and gonadotropin-releasing hormone agonist (GnRHa) to improve the final height of patients.2-5 The rationale for this therapy is to prolong the growth period by inhibiting pubertal progression and delaying epiphyseal fusion,6 although GH deficiency in atrophic thyroiditis patients usually improves after L-thyroxine treatment. We report a case of autoimmune atrophic thyroiditis in a boy who presented with GH deficiency even after his enlarged pituitary gland had decreased in size. Combined treatment with GH, GnRHa, and L-thyroxine was successful in allowing the patient to attain normal adult height.

Background Antisense peptide nucleic acids (PNAs) are synthetic polymers that mimic DNA/RNA and inhibit bacterial gene expression in a sequence-specific manner. Methods To assess activity against non-typeable Haemophilus influenzae (NTHi), we designed six PNA-peptides that target acpP, encoding an acyl carrier protein. MICs and minimum biofilm eradication concentrations (MBECs) were determined. Resistant strains were selected by serial passages on media with a sub-MIC concentration of acpP-PNA. Results The MICs of six acpP-PNA-peptides were 2.9-11 mg/L (0.63-2.5 μmol/L) for 20 clinical isolates, indicating susceptibility of planktonic NTHi. By contrast, MBECs were up to 179 mg/L (40 μmol/L). Compared with one original PNA-peptide (acpP-PNA1-3'N), an optimized PNA-peptide (acpP-PNA14-5'L) differs in PNA sequence and has a 5' membrane-penetrating peptide with a linker between the PNA and peptide. The optimized PNA-peptide had an MBEC ranging from 11 to 23 mg/L (2.5-5 μmol/L), indicating susceptibility. A resistant strain that was selected by the original acpP-PNA1-3'N had an SNP that introduced a stop codon in NTHI0044, which is predicted to encode an ATP-binding protein of a conserved ABC transporter. Deletion of NTHI0044 caused resistance to the original acpP-PNA1-3'N, but showed no effect on susceptibility to the optimized acpP-PNA14-5'L. The WT strain remained susceptible to the optimized PNA-peptide after 30 serial passages on media containing the optimized PNA-peptide. Conclusions A PNA-peptide that targets acpP, has a 5' membrane-penetrating peptide and has a linker shows excellent activity against planktonic and biofilm NTHi and is associated with a low risk for induction of resistance.

Haemophilus influenzae type b (Hib) remains the leading cause of invasive bacterial infection in Japanese children. More than 110 countries that have included Hib conjugate vaccines in their routine vaccination programs have seen dramatical decrease in the incidence of Hib infections. In Japan, the vaccine has been introduced for voluntary immunization since December 2008 and has been provided free of charge only since January 2011. This review reports the prevalence of Hib and its clones among healthy children and pediatric patients diagnosed with invasive or non-invasive Hib infections in Sado Island, Japan. Of 25 Hib isolates collected in this surveillance, 4 genotypic patterns (ST54-gBLPACR-III, ST54-gBLNAR-I/II, ST190-gBLNAS, and ST95-gBLPACR-I/II) were detected. These STs were double or triple-locus variants of each other. Under the same antimicrobial selective pressure, high prevalence of gBLPACR strain (76.0%) was confirmed in Hib isolates, while gBLPACR prevalence in nontypeable H. influenzae was very low (5.2%). These data suggested that each ST strain may be brought into Sado Island by different routes. We note that surveillance of healthy subjects to identify Hib carriers is important to understand their role in transmission of Hib.

Identification of Haemophilus influenzae type b (Hib) in asymptomatic carriers is critical to control the spread of disease. This study was conducted between January 2008 and August 2011 as part of a birth cohort study in Sado Island, Japan, to elucidate the prevalence of Hib and its clones in a specific region. Nasopharyngeal cultures were obtained from 349 subjects at 4-, 7-, 10-, 18-, and 36-month health checkups and analyzed for H. influenzae. The Hib and nontypeable H. influenzae detection rates ranged from 0 to 1.5% (12 isolates) and from 7.9 to 32.9%, respectively. Twelve pediatric patients diagnosed with invasive or non-invasive Hib infections during the study period were also enrolled. The Hib isolates were analyzed for carriage of the beta-lactamase gene and ftsI mutations, and multilocus sequence type (MLST, ST type). Of the 24 Hib isolates, 18 (75%) were ST54, 5 (21%) were ST190, and 1 isolate (4%) was ST95. All of the ST190 isolates were genetically beta-lactamase-negative ampicillin-susceptible isolates, while all but one of the ST54 isolates were genetically beta-lactamase-positive amoxicillin/clavulanic acid-resistant isolates. The geographic distribution of Hib isolates in the study period was scattered. There were 2 day-care cases and 1 family case of Hib infection. The ST54 and ST190 strains circulated in Sado Island and were detected in both asymptomatic carriers and patients. We note that surveillance of healthy subjects to identify Hib carriers is important to understand the transmission of Hib.

Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) strains, which often produce Panton-Valentine leucocidin (PVL), are increasingly noted worldwide. In this study, we examined 42 MRSA strains (25 PVL-positive [PVL + ] strains and 17 PVL-negative [PVL − ] strains) isolated in Taiwan for their molecular characteristics. The PVL + MRSA strains included CA-MRSA strains with multilocus sequence type (ST) 59 (major PVL + MRSA in Taiwan), its variants, and worldwide CA-MRSA ST30 strains. The PVL − MRSA strains included the pandemic Hungarian MRSA ST239 strain, the Hungarian MRSA ST239 variant, MRSA ST59 (largely hospital-acquired MRSA strains) and its variants, the pandemic New York/Japan MRSA ST5 strain (Japanese type), and the MRSA ST8 strain. The major PVL + CA-MRSA ST59 strain possessed a tetracycline resistance-conferring ( tetK positive) penicillinase plasmid and a drug resistance gene cluster (a possible composite transposon) for multidrug resistance. Moreover, it carried a novel staphylococcal cassette chromosome mec (SCC mec ) with two distinct ccrC genes ( ccrC2 - C8 ). This SCC mec (previously named SCC mec type V T ) was tentatively designated SCC mec type VII. Sequencing of the PVL genes revealed the polymorphisms, and the PVL + CA-MRSA ST59 strain possessed the ST59-specific PVL gene sequence. The data suggest that a significant amount of clonal spread is occurring in Taiwan and that the major PVL + CA-MRSA ST59 Taiwan strain exhibits unique genetic characteristics, such as a novel SCC mec type and an ST59-specific PVL gene sequence.

In this study, we report the effectiveness of trivalent inactivated influenza vaccination (TIV) for severely multiply handicapped persons/children (SMHPs) in the 2005-2006 season. In 77 SMHPs, A/New York/55/2004 (H3N2) which was the changed vaccine-strain showed significant differences in the geometric mean titers (P0.05) and seroprotection rates (P0.01) between pre- and post-vaccination. A/New Caledonia/20/99 (H1N1) and B/Shanghai/361/2002, which were the unchanged vaccine-strains, showed no significant differences. We defined the potential responders as those who can achieve 1:40 or more hemagglutination inhibition (HAI) titer after vaccination with any vaccine-strain. Therefore, the rate of potential responders is equivalent to the rate of seroprotection, estimated to be 40-60% among the SMHPs and80% among the control group in this study. In the SMHPs, even potential responders could only achieve limited HAI titers (1:40-80) even after repeated vaccination. In contrast, the control group showed higher HAI titers compared to the SMHPs for the unchanged vaccine-strains caused by the priming effect. These data suggest that it might be difficult for SMHPs (including potential responders) to achieve the priming effect by the current TIV. Consequently, they cannot obtain a booster effect.

The major methicillin-resistant Staphylococcus aureus(MRSA) distributed among hospitals in Japan is New York/Japan clone [multilocus sequence type 5 (ST5), agr type 2 and methicillin resistance locus type (SCC mec) II] which possesses both the toxic shock syndrome toxin 1 gene (tst) and staphylococcal enterotoxin C gene (sec). In this study, we collected 245 MRSA strains from four hospitals during 2001 to 2005 in Niigata, Japan, and analyzed tst and sec genes and SCC mec type among them. A total of 13 strains were further examined for their genotypes, virulence gene patterns and drug resistance. Among the 245 strains four tst sec genes patterns were observed; tst(+) sec(+) strains represented a majority of 86.5% and 9.4% were tst(-) sec(-). SCCmec typing revealed that 91.4% had type II, 4.1% type IV and 4.1% type I. Multilocus sequence typing (MLST) revealed that 10 of the 13 typed strains belonged to clonal complex 5 (7 had ST5 while 3 were single locus variants of ST5) with similar characteristics to the New York/Japan clone and possessed multi-drug resistance with high virulence gene content. The remaining 3 strains were ST8 (n=2) and ST91 (n=1). The ST91 strain had SCC mec IV and seemed to originate in the community, while ST8 strains exhibited SCC mec type I, which is distinct from community type IV. The data suggest that MRSA in hospitals in Niigata now mainly includes the New York/Japan clone (undergoing genomic divergence and clonal expansion) and other minor types (e.g. ST8) as well as the community type.

Staphylococcus aureus and coagulase-negative staphylococci (CNS) isolated from the nasal mucosa of medical students were examined for susceptibility to 16 antimicrobial agents. No methicillin-resistant S. aureus (MRSA) was isolated, while MRCNS was present in 23.5% of the medical students. CNS exhibited significantly more resistance to antimicrobial agents such as gentamicin, in addition to oxacillin, compared to S. aureus, and 13.1% of the CNS strains (mostly MRCNS) were multidrug-resistant (to five or more drugs). In contrast, ampicillin resistance was higher in S. aureus. The rate of hospitalization or of taking an antimicrobial agent within the past 1 year was lower in CNS+ students than in S. aureus+ students. The data suggest that CNS could serve as a reservoir of drug resistance by persistent colonization in the nasal mucosa. In this study, MRCNS with multidrug resistance was found in medical students. More attention should be given to nasal MRCNS in medical students as a possible spreader in hospitals.

Highly virulent, community-acquired methicillin-resistant Staphylococcus aureus (MRSA) strains with Panton-Valentine leucocidin (PVL) genes have been found increasingly worldwide. Among a total of 2,101 MRSA strains isolated from patients in hospitals in Japan, two were positive for PVL genes. One strain was identified as a community-acquired MRSA strain with genotype sequence type 30 (ST30) and spa (staphylococcal protein A gene) type 19 from Japan and was resistant only to β-lactam antimicrobial agents. The other strain was closely related to PVL + multidrug-resistant, hospital-acquired MRSA strains (ST30, spa type 43) derived from nosocomial outbreaks in the 1980s to 1990s in Japan but with a divergent sequence type, ST765 (a single-locus variant of ST30). Twenty-two PVL + MRSA strains, including those from Japan and those from other countries with various sequence types (ST1, ST8, ST30, ST59, and ST80) and genotypes, were examined for susceptibility to 31 antimicrobial agents. Among the agents, DX-619, a des-fluoro(6) quinolone, showed the greatest activity, followed by rifampin and sitafloxacin, a fluoroquinolone. The data suggest that DX-619 exhibits a superior activity against PVL + MRSA strains with various virulence genetic traits from the community as well as from hospitals.

Background Human parechovirus type 3 (HPeV3) and enteroviruses (EVs) are the major viral pathogens causing severe diseases in neonates and young infants. Although two infections typically occur in summer, and initial clinical presentations are similar, differences in clinical and laboratory characteristics of HPeV3- and EV- infections are not well described. Methods From January 2014 to December 2016, we prospectively obtained serum and/or cerebrospinal fluid (CSF) samples from febrile neonates and young infants < 4 months suspecting sepsis and meningoencephalitis in Niigata, Japan. RNA/DNA were extracted from the samples and viral etiologies including HPeV3, EVs, and/or herpes simplex virus were tested using the real-time PCR, followed by genetic sequencing and typing. The comparison was made between HPeV3- and EVs-infected patients based on their medical records and subsequent questionnaires using appropriate statistical analyses. Results In total, we obtained 212 serum and 170 CSF samples from 222 patients. HPeV3 and EVs were detected for 56 (25%) and 43 patients (19%) (Figure), and their median ages were 29 and 24 days (P = 0.58), respectively. HPeV3-infected patients presented with higher median body temperature (38.9 °C vs 38.5 °C, P < 0.001), heart rates (184/minute vs. 161/minute, P = 0.002), peripheral coldness (72% vs. 34%, P < 0.001), skin mottling (65% vs. 23%, P < 0.001), and grunting (22% vs. 5%, P = 0.014) than EVs-infected patients. Additionally, systemic inflammatory response syndrome (SIRS) was more frequently observed in HPeV3-infected patients than in EVs-infected patients (82% vs. 58%, P = 0.009). In the laboratory data, median white blood cell count was lower in HPeV3-infected patients than those in EVs-infected patients (5,200/µL vs. 8,900/µL, P < 0.001). The pleocytosis was observed 58% of EVs-infected patients, but none of the HPeV3-infected patients (P < 0.001); however, HPeV3 RNA was frequently detected than EVs in CSF (90% vs 66%, P = 0.012). Conclusion This study showed that HPeV3-infected neonates and young infants presented more severe clinical manifestations than those infected with EVs. Significant tachycardia, poor peripheral circulation, and fulfilling SIRS criteria at the presentation may be the clues considering HPeV3 infection rather than EVs infection. Disclosures All authors: No reported disclosures.