The Improved Antigen Uptake and Presentation of Dendritic Cells Using Cell-Penetrating D-octaarginine-Linked PNVA-co-AA as a Novel Dendritic Cell-Based Vaccine.

Cancer immunotherapy using antigen-pulsed dendritic cells can induce strong cellular immune responses by priming cytotoxic T lymphocytes. In this study, we pulsed tumor cell lysates with VP-R8, a cell-penetrating D-octaarginine-linked co-polymer of N-vinylacetamide and acrylic acid (PNVA-co-AA), into the DC2.4 murine dendritic cell line to improve antigen uptake and then determined the anti-tumor effect in tumor-bearing mice. DC2.4 cells were pulsed with the cell lysate of EL4, a murine lymphoma cell line, and VP-R8 to generate the DC2.4 vaccine. For the in vivo study, DC2.4 cells pulsed with EL4 lysate and VP-R8 were subcutaneously injected into the inguinal lymph node to investigate the anti-tumor effect against EL4 and EL4-specific T cell immune responses. VP-R8 significantly improved antigen uptake into DC2.4 compared to conventional keyhole limpet hemocyanin (p < 0.05). The expression of MHC class I, MHC class II, and CD86 in DC2.4 cells significantly increased after pulsing tumor lysates with VP-R8 compared to other treatments (p < 0.05). The intra-lymph node injection of DC2.4 pulsed with both VP-R8 and EL4 lysate significantly decreased tumor growth compared to DC2.4 pulsed with KLH and lysates (p < 0.05) and induced tumor-infiltrating CD8T cells. The DC2.4 vaccine also remarkably increased the population of IFN-gamma-producing T cells and CTL activity against EL4 cells. In conclusion, we demonstrated that VP-R8 markedly enhances the efficiency of dendritic cell-based vaccines in priming robust anti-tumor immunity, suggesting its potential as a beneficial additive for dendritic cell-based immunotherapy.

Genetic analysis of ESBL-producing Klebsiella pneumoniae isolated from UTI patients in Indonesia.

INTRODUCTION: Extended spectrum beta-lactamase (ESBL)-producing Klebsiellapneumoniae is a serious concern for nosocomial infection and the emergence rate in Indonesia is higher than that in developed countries. The purpose of this study was to investigate the genetic characteristics of ESBL-producing K. pneumoniae isolated from UTI patients in Indonesia. MATERIALS AND METHODS: We collected K. pneumoniae resistant to ceftazidime or cefotaxime isolated from UTI patients in Dr. Soetomo's Academic Hospital in Surabaya, Indonesia in 2015. Ninety-four strains were identified as ESBL-producing bacteria by confirmation tests. The isolates were investigated by antimicrobial susceptibility testing with 20 drugs and ESBL gene detection, plasmid replicon typing and virulence genes as hypermucoviscous (HMV) strains were tested by the string test. RESULTS: High rates of resistance to ciprofloxacin (86.2%), tetracycline (80.9%) and nalidixic acid (78.7%) were observed. CTX-M-15 was the most common ESBL gene (89.4%), 33 of which also carried SHV-type ESBL. IncF was the most prevalent plasmid replicon typing (47.6%). Sixteen (17.0%) strains were judged as HMV, all of which had rmpA and more than half of which had fimH, uge, and wab. IncL/M was the most common replicon plasmid in the HMV strains, and the difference in the positive rate was statistically significant (p = 0.0024). CONCLUSION: This study showed the high prevalence of multiple-drug resistant and predominately CTX-M-15-positive ESBL-producing K. pneumoniae in Indonesia. There was a correlation between IncL/M and the HMV phenotype in this study. As such hypervirulent strains continue to emerge, studying their dissemination with resistance determinants is an urgent priority.

Cross-Resistance and the Mechanisms of Cephalosporin-Resistant Bacteria in Urinary Tract Infections Isolated in Indonesia.

Urinary tract infection (UTI) by antibiotic-resistant strains has become increasingly problematic, with trends that differ from country to country. This study examined cross-resistance and the mechanisms of cephalosporin resistance in UTI-causative bacteria isolated in Indonesia. Antibiotic susceptibility tests based on Clinical Laboratory Standards Institute (CLSI) standards were done for UTI-causative strains (n = 50) isolated from patients in Indonesia in 2015-2016 and showed resistance against the third-generation cephalosporin. Mechanistic studies were carried out to confirm the presence of extended-spectrum ß-lactamase (ESBL) genes, carbapenemase-related genes, the fosA3 gene related to fosfomycin resistance, and mutations of quinolone-resistance-related genes. Isolated UTI-causative bacteria included Escherichia coli (64.0%), Pseudomonas aeruginosa (16.0%), Klebsiella pneumoniae (10.0%), and others (10.0%). These strains showed 96.0% susceptibility to amikacin, 76.0% to fosfomycin, 90.0% to imipenem, 28.0% to levofloxacin, 92.0% to meropenem, and 74.0% to tazobactam/piperacillin. ESBL was produced by 68.0% of these strains. Mechanistic studies found no strains with carbapenemase genes but 6.0% of strains had the fosA3 gene. Seventy-two % of the strains had mutations in the gyrA gene and 74.0% in the parC gene. Most E. coli strains (87.5%) had Ser-83 → Leu and Asp-87 → Asn in gyrA and 93.8% of E. coli had Ser-80 → Ile in parC. There were significant correlations among mutations in gyrA and parC, and fosA3 gene detection (P < 0.05), respectively. To our knowledge, this is the first mechanistic study of antibiotic-cross-resistant UTI-causative bacteria in Indonesia. Further studies with a longer period of observation are necessary, especially for changes in carbapenem resistance without carbapenemase-related genes.

Differential effects of chromosome and plasmid blaCTX-M-15 genes on antibiotic susceptibilities in extended-spectrum beta-lactamase-producing Escherichia coli isolates from patients with urinary tract infection.

OBJECTIVES: To compare antibiotic susceptibilities between chromosomal and plasmid blaCTX-M-15 locations in urinary tract infection-causing extended-spectrum ß-lactamases-producing Escherichia coli blaCTX-M-15 isolated in Indonesia. METHODS: A total of 84 strains identified as extended-spectrum ß-lactamases-producing E. coli were isolated from patients with urinary tract infection in Indonesia in 2015. Antimicrobial susceptibility tests were performed on these strains using 18 antibiotics, and extended-spectrum ß-lactamase bla genes were detected by polymerase chain reaction. Gene localization of blaCTX-M-15 -positive strains was confirmed by Southern blot hybridization, and epidemiological typing was conducted using multilocus sequence typing. RESULTS: Of 54 strains harboring the blaCTX-M-15 gene, 27 showed localization on chromosome, 20 on plasmid, and seven on chromosome and plasmid. Most multilocus sequence typing sequence types of the 27 strains with chromosomal blaCTX-M-15 were ST405 (25.9%) and ST131 (22.2%) strains, whereas the 20 strains with plasmid-blaCTX-M-15 were mostly ST410 (55.0%). CONCLUSIONS: Extended-spectrum ß-lactamases-producing E. coli blaCTX-M-15 with plasmid genes show significantly higher resistant rates against piperacillin-tazobactam but lower resistant rates against chloramphenicol compared to chromosomal strains in Indonesian patients with urinary tract infection. Mechanistic investigations will be necessary to advance our knowledge of antimicrobial resistance in urinary tract infection.

Can ultrasound irradiation be a therapeutic option for prostate cancer?

BACKGROUND: Focal therapies for prostate cancer (PC) can reduce adverse events and do not lead to androgen-independent progression. Ultrasound could be used for cancer treatments if the repetition frequency is fitted to the purpose. We investigated the possible therapeutic effect of ultrasound irradiation on PC cells. MATERIALS AND METHODS: We irradiated two PC cell lines, androgen-dependent LNCaP and -independent PC-3 with ultrasound (3.0 W/cm2 , 3 MHz, irradiation time rate: 20%) for 2 minutes for 1 day or 3 consecutive days at a repetition frequency of 1, 10, or 100 Hz in vitro. Cell proliferation and apoptosis were determined after irradiation. RESULTS: Cell proliferation of PC-3 was significantly inhibited after 1 day (P < .0001) and 3 days (P < .0001) of 10 Hz ultrasound irradiation, and that of LNCaP after 1 day (P < .0001) and 3 days (P < .0001) of irradiation. LNCaP was more sensitive to ultrasound at both lower and higher cell density but PC-3 was only sensitive at a lower cell density (P < .01). Irradiation with 10 Hz ultrasound-induced significantly more PC-3 apoptotic cells than control (1 day, P = .0137; 3 days, P = .0386) rather than irradiation with 1 Hz. Apoptosis via caspase-3 was induced at 10 Hz in 1-day (P < .05) irradiation in both cell lines. CONCLUSIONS: Ultrasound irradiation with even 1 day of 10 Hz significantly inhibited cell proliferation in both LNCaP and PC-3, especially by the remarkable induction of apoptosis in vitro. Our study indicated that ultrasound irradiation can be a therapeutic option for PC and further studies in vivo will be undertaken.

Risk factors for febrile genito-urinary infection in the catheterized patients by with spinal cord injury-associated chronic neurogenic lower urinary tract dysfunction evaluated by urodynamic study and cystography: a retrospective study.

INTRODUCTION: To investigate the risk factors for febrile genito-urinary tract infection (GUTI) in spinal cord injury-associated neurogenic lower urinary tract dysfunction (NLUTD) patients who perform routine clean intermittent catheterization (CIC) evaluated by urodynamic study (UDS) and cystography. PATIENTS AND METHODS: Over a 3-year period, we retrospectively assessed risk factors for febrile UTI in 141 spinal cord injury patients diagnosed as NLUTD and performing routine CIC, regarding gender, UDS findings such as bladder compliance, maximum cystometric capacity, and cystography. RESULTS: A total of 41 patients had febrile GUTI in the follow-up period as along with 32 cases of pyelonephritis, 10 cases of epididymitis, and 1 case of prostatitis, including patients with multiple infectious diseases. The causative bacteria were Escherichia coli (14 cases) followed by Pseudomonas aeruginosa (n = 5), Klebsiella pneumoniae (n = 4), and Klebsiella oxytoca (n = 4). Antibiotic-resistant E. coli were seen, with 36.4% instances of extended-spectrum beta-lactamase production in whole of E. coli. Male gender (p = 0.018), ASIA Impairment Scale (AIS) C or more severe (p = 0.031), the number of CIC (p = 0.034), use of quinolones (p < 0.001) and severe bladder deformity (DG 2 or more, p = 0.004) were significantly associated with febrile GUTI occurrence. CONCLUSIONS: Our data demonstrated that male gender, severe bladder deformity (DG 2 or more), AIS C or more, the number of CIC, and use of quinolones were significantly associated with febrile GUTI occurrence in NLUTD patients employing routine CIC. Further prospective studies are necessary to define the full spectrum of possible risk factors for febrile GUTI in these patients.

Use of oral third generation cephalosporins and quinolones and occurrence of antibiotic-resistant strains in the neurogenic bladder (NB) outpatient setting: a retrospective chart audit.

STUDY DESIGN: A retrospective chart audit. OBJECTIVES: Neurogenic bladder (NB), a risk factor for urinary tract infection, has not been comprehensively studied in terms of antimicrobial stewardship. In this study, we studied the relationship between the use of oral third generation cephalosporins and quinolones, and the occurrence of antibiotic-resistant strains. SETTING: Hyogo Prefectural Central Rehabilitation Hospital, Hyogo, Japan. METHODS: We retrospectively investigated antibiotic-resistant bacteria and the amount of antibiotics prescribed in outpatients with NB caused by spinal cord injury between 2012 and 2017. We intervened in urological departments whose physicians often prescribed third generation cephalosporins and fluoroquinolone, and analyzed the number of prescriptions and the amount of Cefdinir (CFDN) and Levofloxacin (LVFX), and studied changes of ratios in antibiotic-resistant strains such as extended-spectrum ß-lactamases (ESBLs) and quinolone-resistant Escherichia coli and Klebsiella pneumoniae. RESULTS: The number of CFDN prescriptions per year significantly decreased from 463 cases to 130 cases over 6 years (p = 0.012). The number of LVFX prescriptions per year decreased from 640 cases to 171 cases (p = 0.025). The incidence rate of ESBL-producing K. pneumoniae decreased from 25% to 7% of total K. pneumoniae (p < 0.001). The incidence of LVFX-resistant E. coli and K. pneumoniae significantly decreased in 2017 compared with 2012 (p = 0.03 and p = 0.016, respectively). CONCLUSIONS: Antimicrobial stewardship interventions decreased the use of CFDN and LVFX for outpatients with NB. Our findings suggested that the reduction in the use of third generation cephalosporins and quinolones correlates with observed decrease in the occurrence of antibiotic-resistant, ESBL-producing, and quinolone-resistant bacteria.

Effect of Preoperative Dutasteride on Holmium Laser Enucleation of the Prostate.

PURPOSE: The aim of this study was to compare and investigate the efficacy of using the 5α-reductase inhibitor dutasteride after holmium laser enucleation of the prostate (HoLEP) to improve postoperative urination and surgery-related complications. METHODS: This is a retrospective observational study comparing patients who received or did not receive 5α-reductase inhibitors prior to HoLEP. Of a total of 270 patients, 40 received the 5α-reductase inhibitor dutasteride. We compared the factors including age, postoperative maximal flow rate (MFR; mL/s), postoperative prostate-specific antigen (PSA) (ng/mL), preoperative MFR (mL/s), preoperative PSA (ng/mL), prostate cancer (%), operative time (min), preoperative residual urine (mL), postoperative residual urine (mL), urinary incontinence (day 1; %), urinary incontinence (1 month; %), urinary incontinence (3 months; %), urethral catheter indwelling period (days), morcellation time (min), enucleation time (min), intraoperative complications (%), postoperative complications (%), prostate volume (mL), enucleated weight (g), and hospitalization period (days). RESULTS: Postoperative PSA (p = 0.0071), morcellation time (p = 0.0444), postoperative complications (p = 0.0350) and prostate volume (p = 0.0069), but not enucleated prostate weight (p = 0.8809), were significantly lower in the dutasteride group. Importantly, enucleation efficiency and morcellation efficiency did not show any significant difference between the dutasteride and the non-dutasteride groups. CONCLUSIONS: Use of a preoperative 5α-reductase inhibitor significantly correlated with surgery-related factors, with less morcellation time, fewer postoperative complications, and lower postoperative PSA. Surgeons performing HoLEP may wish to take these findings into account.

Difference of Phenotype and Genotype Between Human and Environmental: Isolated Vibrio cholerae in Surabaya, Indonesia.

Cholera due to Vibrio cholerae has been spreading worldwide, although the reports focusing on Indonesian V. cholerae are few. In this study, in order to investigate how V. cholerae transmitted to human from environment. We extended an epidemiological report that had investigated the genotype of V. cholerae isolated from human pediatric samples and environmental samples. We examined 44 strains of V. cholerae isolated from pediatric diarrhea patients and the environment such as shrimps or oysters collected in three adjacent towns in Surabaya, Indonesia. Susceptibilities were examined for 11 antibiotics. Serotype O1 or O139 genes and pathogenic genes including cholera toxin were detected. Multi-locus sequence typing (MLST) and enterobacterial repetitive intergenic consensus (ERIC)-PCR were also performed to determine genetic diversity of those isolates. Serotype O1 was seen in 17 strains (38.6%) with all pathogenic genes among 44 isolates. Other isolates were non-O1/non-O139 V. cholerae. Regarding antibiotic susceptibilities, those isolates from environmental samples showed resistance to ampicillin (11.4%), streptomycin (9.1%) and nalidixic acid (2.3%) but those isolates from pediatric stools showed no resistance to those 3 kinds of antibiotics. MLST revealed sequence type (ST) 69 in 17 strains (38.6%), ST198 in 3 strains (6.8%) and non-types in 24 strains (54.5%). All the ST69 strains were classified to O1 type with more than 95% similarity by ERIC-PCR, including all 6 (13.6%) isolates from environmental samples with resistance to streptomycin. In conclusion, V. cholerae O1 ST69 strains has been clonally spreading in Surabaya, exhibiting pathogenic factors and antibiotic resistance to streptomycin, especially in the isolates from environment.

Human herpesvirus 6 encephalitis in patients administered mycophenolate mofetil as prophylaxis for graft-versus-host disease after allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Human herpesvirus 6 (HHV-6) encephalitis is a known life-threatening complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, few studies have focused on the occurrence of HHV-6 encephalitis in patients receiving mycophenolate mofetil (MMF) combined with a calcineurin inhibitor as prophylaxis for graft-versus-host disease (GVHD). This study aimed to investigate the impact of MMF administered for GVHD prophylaxis in the occurrence of HHV-6 encephalitis after allo-HSCT and the characteristics of this condition. METHODS AND RESULTS: We retrospectively analyzed 73 patients who underwent allo-HSCT (83 transplants) at our hospital between April 2010 and December 2015. MMF (2-3 g/d) was administered along with a calcineurin inhibitor. Seven patients (8.0%) developed encephalitis due to HHV-6. The median period from allo-HSCT to the onset of HHV-6 encephalitis was 23 days (range, 17-98 days). The cumulative incidence of HHV-6 encephalitis on day 100 after treatment was 12% and 6% in patients who underwent cord blood transplantation (CBT) and non-CBT (ie, bone marrow transplantation and peripheral blood stem cell transplantation), respectively (P = 0.344). Neurological symptoms of encephalitis were more severe in non-CBT cases than those in CBT cases. All patients diagnosed with HHV-6 encephalitis were treated with ganciclovir or foscarnet. None of the enrolled patients died from HHV-6 encephalitis. CONCLUSIONS: Mycophenolate mofetil may have the potential to increase the frequency of severe HHV-6 encephalitis in patients undergoing CBT and non-CBT. Thus, MMF should be administered with caution, and patients should be monitored closely for HHV-6 encephalitis even those who did not undergo CBT.

Clinical Risk Factors for Death Caused by Extended-Spectrum Beta-Lactamase: Producing Bacteria.

OBJECTIVES: Extended-spectrum beta-lactamase (ESBL)-producing bacteria often causes bacteremia, leading serious outcomes. In this study, we conducted a retrospective analysis to identify the risk factors associated with death by bacteremia of ESBL-producing bacteria. METHODS: Patients with bacteremia by ESBL-producing bacteria were retrospectively collected in Kobe University Hospital, Japan, between January 2011 and December 2015. Potential risk factors for death caused by ESBL-bacteremia were analyzed for patients' outcome (recovery or death) by univariate and multivariate analysis. RESULTS: A total of 101 patients (64 male and 37 female) were recruited. The most frequently detected ESBL-producing bacteria were Escherichia coli (91 cases; 90.1%), followed by Klebsiella pneumoniae (8 cases; 7.9%). Most frequently used antibiotics after the detection of bacteremia was meropenem (66.3%; 67/101) followed by cefmetazole (51.5%; 52/101). Univariate analysis showed a significantly positive correlation with mortality in ICU admission (p < 0.001), circulatory diseases (p = 0.022), shock (p = 0.044), and respirator requirement (p = 0.002). Multivariate analysis showed ICU admission remained significant risk factor for mortality (p = 0.0192). CONCLUSIONS: We showed ICU admission was significantly correlated with death from bacteremia by ESBL-producing bacteria. These factors should be monitored to estimate severity of ESBL causing-bacteremia for better patients' outcomes.

Molecular characteristics of carbapenem-resistant Pseudomonas aeruginosa isolated from urine in Hyogo, Japan.

OBJECTIVES: To investigate the molecular characteristics and epidemiology of metallo-ß-lactamase-producing Pseudomonas aeruginosa from urine of urinary tract infection patients in Hyogo Prefecture, Japan. METHODS: Carbapenem-resistant P. aeruginosa isolated from the urine of 21 urinary tract infection patients in three general hospitals in Hyogo Prefecture (Japan) were collected between 2007 and 2014. Their antibiotic susceptibilities, metallo-ß-lactamase screening test, metallo-ß-lactamase gene sequencing, multilocus sequence typing and repetitive-sequence-based polymerase chain reaction were determined for epidemiological analyses to investigate the genetic characteristics. RESULTS: Out of 21 isolates, 13 (61.9%) were positive for metallo-ß-lactamase. There were 11 (52.4%) isolates with IMP-1 in them, one (4.5%) isolate with IMP-7 and one (4.5%) isolate with VIM-1. Metallo-ß-lactamase-positive isolates were mainly identified as ST235, and metallo-ß-lactamase-negative isolates were STs 357, 277, 234, 439 and 639. Repetitive-sequence-based polymerase chain reaction showed metallo-ß-lactamase-positive isolates were grouped in eight clusters, and ST235 isolates with IMP-1 from three hospitals belonging to the identical group I, the other ST235 isolates with IMP-7 and VIM-1 were from two hospitals belonging to group II. CONCLUSIONS: Metallo-ß-lactamase-positive P. aeruginosa of ST235 isolates with IPM-1 were mainly identified from the urine of urinary tract infection patients in Hyogo, Japan. A ST235 isolate with VIM-1 was found for the first time. Further investigation is necessary to follow the spread of metallo-ß-lactamase-positive isolates.

Bacteremia complicating urinary tract infection by Pseudomonas aeruginosa: Mortality risk factors.

OBJECTIVES: To examine the clinical risk factors for death within 30 days of diagnosis of Pseudomonas aeruginosa-causing bacteremia after a urinary tract infection. METHODS: A total of 62 patients with Pseudomonas aeruginosa isolated from both urine and blood at the same episode from January 2009 to December 2016 were enrolled in the present study. We retrospectively investigated clinical risk factors for death by comparison between surviving patients and those who died within 30 days after diagnosis of P. aeruginosa bacteremia. The comparison for risk factors for bacteremia-related death included 31 categories, such as age, laboratory data, underlying diseases, clinical history, history of surgery, care in the intensive care unit, P. aeruginosa susceptibility to the antibiotics used at the time of bacteremia diagnosis and consultation with urological department. RESULTS: The study included 48 men and 14 women aged 71.3 ± 10.4 years. Nine patients (14.5%) died of P. aeruginosa bacteremia. Statistical analysis showed that non-survivors had significantly lower albumin levels than survivors (2.07 ± 0.62 vs 2.62 ± 0.65; P = 0.023). The non-survivors had significantly higher rates of ventilator use, history of heart disease, septic shock and lower rates of consultation with urological departments after diagnosis (P < 0.05). CONCLUSIONS: Patients with bacteremia complicating urinary infection by P. aeruginosa have a low death rate. Earlier intervention by urologists might improve patients' outcome. Lower albumin levels, ventilator use, history of heart disease and septic shock are factors associated with higher mortality rate.

Prevalence of Quinolone Resistance of Extended-Spectrum ß-Lactamase-Producing Escherichia coli with ST131-fimH30 in a City Hospital in Hyogo, Japan.

Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli isolates are known to tolerate superior quinolone antimicrobials compared with other antibacterial agents. Among the clones belonging to sequence type (ST) 131 by multilocus sequence typing, the involvement of the H30-Rx subclone has been reported worldwide with various fimH genes encoding type 1 pili. We investigated 83 isolates of ESBL-producing E. coli and performed antimicrobial susceptibility test, CH (fumC/fimH) ST131 by typing the specific PCR. Moreover, mutation analysis of genes involved in quinolone antibiotic resistance (gyrA and parC) and ESBL genotypes were determined. As a result, 54 of 83 isolates (65.1%) of CH40-30 clones corresponding to ST131-fimH30 were detected, and all were resistant to levofloxacin. Mutations associated with this resistance were common, and included S83L and D87N of gyrA and S80I and E84V of parC. Subclone analysis revealed a high proportion of fimH30-non-Rx (40 isolates, 74.1%). Each subclone was characterized by ESBL genotype, and the CTX-M-15 type was mainly seen for fimH30-Rx, with the CTX-M-14 type or CTX-M-27 type seen for fimH30-non-Rx. This study suggests that an increase in ESBL-producing quinolone-resistant E. coli in a city hospital in Hyogo, Japan, was caused by the spread of subclones belonging to fimH30-non-Rx of ST131.

Risk factors for death from Stenotrophomonas maltophilia bacteremia.

PURPOSE: Stenotrophomonas maltophilia has low pathogenicity potential, but if it causes bacteremia it can be fatal, because it has shown high resistance to many antibiotics and can be difficult to treat. Patient death from S. maltophilia bacteremia has increased since 2014 in our hospital. In this study, we investigated risk factors for death due to S. maltophilia bacteremia. METHODS: Seventy patients from the hospital database with S. maltophilia bacteremia between January 2010 and July 2017 were investigated. We retrospectively analyzed risk factors including gender, age, wards, hospitalized duration, clinical history, devices, source of S. maltophilia identification, polymicrobial bacteremia, prior antimicrobial therapy, antimicrobial therapy after bacteremia, and resistance to antibiotics. The statistical analysis was performed to compare the period from 2010 to 2013 to from 2014 to 2017. RESULTS: Comparing the 2010-2013 period to the 2014-2017 period, it revealed that history of hospitalization, identification of S. maltophilia from sputum, polymicrobial bacteremia, prior carbapenem use, and mortality was significantly different in S. maltophilia bacteremia (p = 0.028, p = 0.004, p < 0.001, p = 0.034, and p = 0.007, respectively). Comparison between non-survivors and survivors for 2010-2013 and 2014-2017 found ICU admission and ventilator use were seen more often in non-survivors (p = 0.030 vs p = 0.013 and p = 0.027 vs p = 0.010, respectively). CONCLUSIONS: Our analyses showed increase in mortality from S. maltophilia bacteremia from 2014 to 2017, and that non-survivors had a higher frequency of ICU admission and ventilator use in both the 2010-2013 and 2014-2017 periods. There were more combination antimicrobial therapy cases after bacteremia in 2014-2017. Further prospective studies with larger numbers of patients should be undertaken for definitive conclusions.

Comparison between non-septic and septic cases in stone-related obstructive acute pyelonephritis and risk factors for septic shock: A multi-center retrospective study.

PURPOSE: This study assessed risk factors for septic shock in patients with obstructive acute pyelonephritis (APN) associated with upper urinary tract calculi in a multi-center retrospective study. METHODS: We studied 143 patients admitted to 4 hospitals in Japan with obstructive APN associated with upper urinary tract calculi. Data on gender, age, hypertension, diabetes, neurological disease or malignant disease, laboratory data (white blood cell (WBC) and C-reactive protein (CRP)), drainage, and bacterial strains including Escherichia coli in the non-septic and septic groups were collected. Risk factors for septic shock were analyzed by univariate and multivariate statistical analyses. RESULTS: There were a total of 107 non-septic cases (74.8%) and 36 septic cases (25.2%). The commonest strains of urinary tract infection-causative bacteria were E. coli in the non-septic group (23 cases, 21.5%) and septic group (13 cases, 36.1%) (p > 0.05). Emergency drainage was administered in 74.8% of the non-septic group and 97.2% of the septic group (p > 0.05). Meropenem was most often used as the initial treatment in the non-septic group (20 cases, 18.7%) and septic group (22 cases, 61.1%) (p < 0.0001). Risk factors for septic shock in multivariate analyses were diabetic mellitus (odds ratio (OR) = 3.591, p = 0.0098) and CRP ≥ 10 (OR = 1.057, p = 0.0119) as significant independent factors in this multicenter study. CONCLUSIONS: APN is a common infectious disease, especially in the cases with urinary tract obstruction where patients easily acquire bacteremia or sepsis. Stone-associated obstructed APN can cause fatal septic shock in cases with diabetes and CRP ≥ 10. Further prospective studies will be undertaken to draw definitive conclusions.

Molecular epidemiologic study of Clostridium difficile infections in university hospitals: Results of a nationwide study in Japan.

We conducted a nationwide molecular epidemiological study of Clostridium difficile infection (CDI) in Japan investigated the correlation between the presence of binary toxin genes and CDI severity. This is the first report on molecular epidemiological analyses for CDI in multiple university hospitals in Japan, to our knowledge. We examined 124,484 hospitalized patients in 25 national and public university hospitals in Japan between December 2013 and March 2014, investigating antimicrobial susceptibilities and toxin-related genes for C. difficile isolates from stools. Epidemiological genetic typing was performed by PCR-ribotyping and repetitive sequence-based (rep)-PCR to examine the genetic similarities. The results detected toxin A-positive, toxin B-positive, binary toxin-negative (A+B+CDT-) detected from 135 isolates (80.8%) and toxin A-negative, toxin B-positive, binary toxin-negative (A- B+CDT-) in 23 (13.8%). Toxin A-positive, toxin B-positive, and binary toxin-positive (A+B+CDT+) were seen in 9 isolates (5.4%). Vancomycin (n = 81, 37.7%) or metronidazole (n = 88, 40.9%) therapies were undertaken in analyzed cases. Ribotypes detected from isolates were 017/subgroup 1, 070, 078, 126, 176, 449, 475/subgroup 1, 499, 451, 566 and newtypes. Rep-PCR classified 167 isolates into 28 cluster groups including 2-15 isolates. In addition, 2 pairs of strains isolated from different institutions belonged to the same clusters. Seven out of 9 (77.8%) of the patients with binary toxin producing strains had "mild to moderate" outcome in evaluated symptoms. In conclusion, we found that binary toxin did not show regional specificity and had no relevance to severity of CDI.

Improved bacterial identification directly from urine samples with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

BACKGROUND: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) contributes to rapid identification of pathogens in the clinic but has not yet performed especially well for Gram-positive cocci (GPC) causing complicated urinary tract infection (UTI). The goal of this study was to investigate the possible clinical use of MALDI-TOF MS as a rapid method for bacterial identification directly from urine in complicated UTI. METHODS: MALDI-TOF MS was applied to urine samples gathered from 142 suspected complicated UTI patients in 2015-2017. We modified the standard procedure (Method 1) for sample preparation by adding an initial 10 minutes of ultrasonication followed by centrifugation at 500 g for 1 minutes to remove debris such as epithelial cells and leukocytes from the urine (Method 2). RESULTS: In 133 urine culture-positive bacteria, the rate of corresponded with urine culture in GPC by MALDI-TOF MS in urine with standard sample preparation (Method 1) was 16.7%, but the modified sample preparation (Method 2) significantly improved that rate to 52.2% (P=.045). Method 2 also improved the identification accuracy for Gram-negative rods (GNR) from 77.1% to 94.2% (P=.022). The modified Method 2 significantly improved the average MALDI score from 1.408±0.153 to 2.166±0.045 (P=.000) for GPC and slightly improved the score from 2.107±0.061 to 2.164±0.037 for GNR. CONCLUSION: The modified sample preparation for MALDI-TOF MS can improve identification accuracy for complicated UTI causative bacteria. This simple modification offers a rapid and accurate routine diagnosis for UTI, and may possibly be a substitute for urine cultures.

Occurrence and characterization of carbapenem-resistant Gram-negative bacilli: A collaborative study of antibiotic-resistant bacteria between Indonesia and Japan.

OBJECTIVES: To explore the occurrence and characterization of carbapenemase-producing pathogens among carbapenem-resistant Gram-negative bacilli isolated from hospitalized patients with urinary tract infection in Indonesia. METHODS: This was a study promoted by the Japanese-Indonesian collaborative research program in the Japan Initiative for Global Research Network on Infectious Diseases. Bacterial pathogens were prospectively isolated from urine specimens of hospitalized urinary tract infection patients at Dr. Soetomo Hospital (Surabaya, Indonesia). All Gram-negative bacteria resistant to third-generation cephalosporin or carbapenem were included in this study. Carbapenemase genes were investigated for phenotype and genotype. RESULTS: In total, 1082 Gram-negative bacilli were isolated, of which 116 strains were resistant to imipenem or meropenem (carbapenem-resistant Gram-negative bacilli), and 22 strains were carbapenemase-producing Gram-negative bacilli. Carbapenemase-producing Gram-negative bacilli consisted of Acinetobacter baumannii (n = 4), Pseudomonas aeruginosa (n = 4), Klebsiella pneumoniae (n = 5), Providencia rettgeri (n = 4) and five others. The carbapenemase-producing Gram-negative bacilli included NDM-1 (n = 18, 81.8%, in Enterobacteriaceae and Acinetobacter spp.) and IMP-7 (n = 4, 18.2%, all in P. aeruginosa). Among carbapenem-resistant Gram-negative bacilli, all four P. aeruginosa were sensitive to colistin, and all six Acinetobacter spp. were sensitive to minocycline, colistin and tigecycline. Of those patients harboring carbapenemase-producing Gram-negative bacilli, 12 (54.5%) were seriously ill at the time of admission, with longer hospital stays and three deaths (13.6% mortality rate). CONCLUSIONS: Urinary tract infection-causing carbapenem-resistant Gram-negative bacilli are widely disseminated in Indonesia. The NDM-1 phenotype seems to be dominant, and it can be treated with colistin and tigecycline in most cases. Most patients harboring carbapenemase-producing Gram-negative bacilli are seriously ill, have a bad prognosis, with a longer hospital stay and a significant mortality rate.

Development of oral cancer vaccine using recombinant Bifidobacterium displaying Wilms' tumor 1 protein.

Several types of vaccine-delivering tumor-associated antigens (TAAs) have been developed in basic and clinical research. Wilms' tumor 1 (WT1), identified as a gene responsible for pediatric renal neoplasm, is one of the most promising TAA for cancer immunotherapy. Peptide and dendritic cell-based WT1 cancer vaccines showed some therapeutic efficacy in clinical and pre-clinical studies but as yet no oral WT1 vaccine can be administrated in a simple and easy way. In the present study, we constructed a novel oral cancer vaccine using a recombinant Bifidobacterium longum displaying WT1 protein. B. longum 420 was orally administered into mice inoculated with WT1-expressing tumor cells for 4 weeks to examine anti-tumor effects. To analyze the WT1-specific cellular immune responses to oral B. longum 420, mice splenocytes were isolated and cytokine production and cytotoxic activities were determined. Oral administrations of B. longum 420 significantly inhibited WT1-expressing tumor growth and prolonged survival in mice. Immunohistochemical study and immunological assays revealed that B. longum 420 substantially induced tumor infiltration of CD4+T and CD8+T cells, systemic WT1-specific cytokine production, and cytotoxic activity mediated by WT1-epitope specific cytotoxic T lymphocytes, with no apparent adverse effects. Our novel oral cancer vaccine safely induced WT1-specific cellular immunity via activation of the gut mucosal immune system and achieved therapeutic efficacy with several practical advantages over existing non-oral vaccines.