Molecular characteristics and antimicrobial susceptibility of penicillinase-producing Neisseria gonorrhoeae isolates in Fukuoka, Japan, 1996-2018.

OBJECTIVES: The objective of this study was to investigate the molecular characteristics and antimicrobial susceptibility of penicillinase-producing Neisseria gonorrhoeae (PPNG) isolates collected in Fukuoka, Japan, from 1996-2018. METHODS: Antimicrobial susceptibility to seven antibiotics was determined by the agar dilution method. Molecular characteristics were determined by Sanger sequencing of the blaTEM allele, plasmid typing and N. gonorrhoeae multiantigen sequence typing (NG-MAST). Furthermore, full sequences of the penA gene, encoding penicillin-binding protein 2 (PBP2), of PPNG isolates with reduced susceptibility to cefixime were analysed. RESULTS: Among 50 PPNG isolates, 17 and 33 were collected during 1996-2006 and 2007-2018, respectively. In 1996-2006, blaTEM-1 in African plasmid was most frequent (64.7%), followed by blaTEM-1 in Asian plasmid (29.4%) and blaTEM-135 in Toronto/Rio plasmid (5.9%). In 2007-2018, blaTEM-135 in Toronto/Rio plasmid was predominant (54.5%), followed by blaTEM-1 in African plasmid (36.4%) and blaTEM-135 in Asian plasmid (6.1%). Among isolates with the blaTEM-135-carrying Toronto/Rio plasmid in 2007-2018, a novel genogroup G15576 was predominant (66.7%). Isolates with the TEM-135 ß-lactamase were more resistant to ciprofloxacin but were more susceptible to ceftriaxone and tetracycline than isolates with TEM-1. Seven PPNG isolates less susceptible to cefixime possessed the plasmidic blaTEM-1 allele and had mosaic or non-mosaic alterations within PBP2. CONCLUSION: The proportion of PPNG with the blaTEM135-carrying Toronto/Rio plasmid increased during the last 12 years. The increase in PPNG carrying the blaTEM-135 allele is of particular concern as it is considered a possible direct precursor of an extended-spectrum ß-lactamase (ESBL).

[COMBINING MODALITY THERAPY WITH SURGERY CAN PROVIDE LONG-TERM SURVIVAL WITH IMPROVED QUALITY OF LIFE FOR PATIENTS WHO HAVE PROSTATE CANCER WITH INTRACRANIAL DURAL METASTASIS: A CASE REPORT].

A 61-year-old man visited our hospital with a headache and left visual field defect. A head MRI showed an intracranial dural tumor with cerebral compression, which was suspected to be metastatic. Analysis of the tumor markers revealed an increase in prostate-specific antigen (PSA) levels (172.8 ng/mL), and therefore prostate cancer was suspected as the primary tumor. Histological diagnosis of a prostatic tissue sample using a transrectal needle biopsy gave a prostate carcinoma with Gleason score of 5+4=9. Additional imaging examinations revealed metastatic lesions in the intra-pelvic lymph node and bones. These data indicated to us that curative surgery was unlikely to be successful, but finally we decided to perform a craniotomy for tumor resection for the intracranial dural tumor to remove his neurological symptoms. After surgery, his headache and visual field defect improved. The pathological finding was intracranial dural metastasis from prostate cancer and the clinical stage was diagnosed as T3bN1M1c in the UICC criteria (ver. 8). Endocrine therapy with degarelix and bicalutamide was started for the primary and residual metastatic prostate cancers. After one year of initial treatment, bicalutamide was changed to enzalutamide because of a tendency towards increased plasma PSA levels. The patient has survived for two and a half years after surgery with no new metastatic tumors or intracranial tumors. Our experience indicates that combined modality therapy with surgery can provide long-term survival with no cranial nerve disorders for patients who have prostate cancer with intracranial dural metastasis.

Role of Computed Tomography Volumetry in Preoperative Donor Renal Function Evaluation of Living Related Kidney Transplantation.

INTRODUCTION: Renal scintigraphy is used to evaluate split renal function. A computed tomography (CT) examination is also carried out for donor safety and appropriate transplantation surgery, and the renal volume (CT volumetry) can be obtained at that time. In this study, we evaluated donor kidney function by inulin clearance (Cin) before and after donor nephrectomy in living donor renal transplantation, and the predictive role of CT volumetry was compared with diethylenetriamine pentaacetic acid (DTPA). METHOD: From November 2005 to April 2018, 34 cases of living donor transplantation conducted at Fukuoka University Hospital were retrospectively studied. The donated kidney weight was measured in 25 cases, and postoperative Cin was measured in 19 cases. RESULTS: The average donor age was 51.7 years old (from 35 to 71). Preoperative Cin and postoperative Cin of donors were 86.3 mL/min/1.73 m2 (from 59.5 to 138.3) and 52.3 (from 40.5 to 76.6), respectively. The average CT volumetry of donated kidneys was 153.9 mL (from 107.8 to 219.3). Correlations of weight and DTPA and CT volumetry of donated kidneys were r = 0.033 (P = .8770) and r = 0.763 (P < .0001), respectively. Correlations of glomerular filtration rate of DTPA and CT volumetry and Cin of postoperative donor residual kidneys were r = 0.66 (P = .002) and r = 0.555 (P = .014). CONCLUSION: There was a significant correlation between CT volumetry and the weight of the removed kidneys, and a correlation between Cin after donor nephrectomy and CT volumetry of the remaining kidneys, but it did not exceed the predictive role of DTPA. However, it was suggested that it is worthy to use as a preoperative examination for split renal function equivalent to DTPA.

Antimicrobial resistance and molecular characterisation of Neisseria gonorrhoeae isolates in Fukuoka, Japan, 1996-2016.

OBJECTIVES: Antimicrobial resistance and molecular characteristics of Neisseria gonorrhoeae isolates obtained from 1996-2005 (n=200) and 2008-2016 (n=200) in Fukuoka, Japan, were examined. METHODS: MICs were determined by agar dilution. Sequence types (STs) were examined using N. gonorrhoeae multiantigen sequence typing (NG-MAST). Sequencing of major extended-spectrum cephalosporin (ESC) resistance determinants (penA, mtrR and ponA) was performed. RESULTS: Increases in the proportion of gonococci with decreased susceptibility or resistance to cefixime (from 18.0% in 1996-2005 to 46.0% in 2008-2016) and ceftriaxone (from 2.5% to 4.0%) were observed. Gonococcal isolates also showed increased resistance to ciprofloxacin and azithromycin. The four most prevalent NG-MAST STs with a multidrug-resistant phenotype were ST2958 (n=18), ST1407 (n=14), ST6798 (n=12) and ST4015 (n=10). The number of isolates belonging to these four STs rose between the first and second period. Among the 54 isolates belonging to the four major STs, 42 (77.8%) contained a penA mosaic allele and 12 (22.2%) contained a penA non-mosaic allele. The sequence pattern types in the 42 isolates with a penA mosaic allele included type X (64.3%), type XXXIV (33.3%) and a novel pattern type (2.4%). In contrast, all 12 isolates with the penA non-mosaic allele included the sequence pattern type V. CONCLUSION: Neisseria gonorrhoeae isolates with decreased susceptibility or resistance to ESC have increased over the years. Four major STs with a multidrug-resistant phenotype were identified. These isolates contained a penA mosaic allele or a non-mosaic allele.

High Prevalence of Azithromycin-Resistant Neisseria gonorrhoeae Isolates With a Multidrug Resistance Phenotype in Fukuoka, Japan.

BACKGROUND: The current guidelines recommend a combination of ceftriaxone and azithromycin as a first-line treatment of gonorrhea in the United States and Europe. Despite not being recommended as a first-line regimen in Japan, an oral 2-g dose of azithromycin did become available for gonococcal infections in 2009. Recently, the emergence of azithromycin-resistant Neisseria gonorrhoeae isolates has been reported in several countries, including Japan. METHODS: Antimicrobial susceptibility testing was performed on a total of 677 clinical isolates of N. gonorrhoeae obtained from January 2010 to December 2013 in Fukuoka, Japan. A molecular analysis by N. gonorrhoeae multiantigen sequence typing was conducted on the azithromycin-resistant isolates. RESULTS: The proportion of azithromycin-resistant isolates (minimum inhibitory concentration > 0.5 µg/mL) increased significantly from 1.8% in 2010 to 22.6% in 2013 (P < 0.001). Among 50 azithromycin-resistant isolates, 30 (60%) exhibited a resistant phenotype to multiple drugs including cefixime. The 2 predominant sequence types (STs) identified by N. gonorrhoeae multiantigen sequence typing were ST6798 (por allele 4033 and tbpB allele 110) and ST1407 (por allele 908 and tbpB allele 110) at 40.0% (20/50) and 12.0% (6/50), respectively. There was a statistically significant increase of the proportion of ST6798 from 0% (0/19) in 2010-2012 to 64.5% (20/31) in 2013 (P < 0.001). CONCLUSIONS: Over the previous 4 years, an increasing prevalence of azithromycin-resistant N. gonorrhoeae isolates with a multidrug-resistant phenotype was observed. Furthermore, the azithromycin-resistant isolates seemed to belong to 2 predominant STs. As a result, continued surveillance of gonococci resistant to antimicrobial agents, including azithromycin in Fukuoka, Japan, is necessary.

In vitro activities of antimicrobial combinations against clinical isolates of Neisseria gonorrhoeae.

The Centers for Disease Control and Prevention (CDC) now recommend combination therapy with ceftriaxone 250 mg plus azithromycin (AZM) 1 g as a first-line regimen for gonorrhea because the increase of Neisseria gonorrhoeae resistant to multiple antimicrobial agents. However, reports on the in vitro activity of antimicrobial combinations against clinical isolates of N. gonorrhoeae are very rare. In the present study, a checkerboard method was utilized to examine the in vitro activity of ceftriaxone (CTRX), cefodizime (CDZM), spectinomycin (SPCM), or gentamicin (GM) in combination with AZM against 25 clinical isolates of N. gonorrhoeae. The SPCM + AZM combination demonstrated the lowest mean fractional inhibitory concentration index (FICI) of 0.69, followed by the CDZM + AZM combination (mean FICI, 0.75), the CTRX + AZM combination (mean FICI, 0.81), and the GM + AZM combination (mean FICI, 0.83). Additivity/indifference effect was detected for the SPCM + AZM combination, the CDZM + AZM combination, the CTRX + AZM combination, and the GM + AZM combination, against 96%, 72%, 92%, and 100% of the isolates, respectively. There was no antagonism for any of the antimicrobial combinations against the 25 N. gonorrhoeae isolates. These results suggest that the antimicrobial combinations may be worthy of clinical evaluation as an alternative regimen for gonococcal infections caused by antimicrobial-resistant strains.

[Neisseria gonorrhoeae infections].

Neisseria gonorrhoeae infections are common bacterial sexually transmitted diseases. Men will usually experience lower urinary tract symptons attributed to urethritis, epididymitis, proctitis, or prostatitis, with associated mucopurulent urethral discharge. Many women are asymptomatic. But, occasionally, they have symptons of vaginal and pelvic discomfort of dysuria, and these infections can lead to pelvic inflammatory disease. Recentry, high prevalence of Neisseria gonorrhoeae isolates resistant to antimicrobial agents is a serious problem in the treatment of gonorrhea. For example, in Fukuoka city, Japan, the proportion of the isolates resistant to ciprofloxacin (CPFX) were 73.4% in 2006 and it was still so high. The proportion of the isolates resistant to tetracycline (TC) was 38.5% in 2006 and that of isolates resistant to penicillin G (PCG) was 17.5%. Owing to this high prevalence of antimicrobial-resistant Neisseria gonorrhoeae in Japan, the clinical efficacy rates of oral antimicrobial agents have become lower. So, as first-line therapy for gonococcal infections, only three parenteral regimens of single doses of ceftriaxone, cefodizime or spectinomycin are recommended by the Japanese Society for Sexually Transmitted Diseases. In the circumstances, we studied in vitro activity of combinations of oral agents such as, beta-lactam and azithromycin, fluoroquinolone and azithromycin, or beta-lactam and fluoroquinolone against Neisseria gonorrhoeae. The cefixime+azithromycin combination demonstrated greater synergy than other combinations.

Antimicrobial resistance in clinical isolates of Neisseria subflava from the oral cavities of a Japanese population.

A recent study indicated that Neisseria subflava, one of the commensal Neisseria species, may play an important role in the emergence of Neisseria gonorrhoeae strains with chromosomally mediated resistance to penicillin or cephalosporin by the horizontal genetic exchange of penA genes encoding the target site for penicillin or cephalosporin. The present investigation examined the antimicrobial susceptibility of 45 isolates of N. subflava from the oral cavities of Japanese men and women to various agents used for the treatment of gonococcal infections. Of the 45 isolates, 40 (88.9%) and 4 (8.8%) were intermediately resistant and resistant to penicillin, respectively, with the minimal inhibitory concentration (MIC)(50) and MIC(90) of penicillin being 0.5 mg/l and 1 mg/l, respectively. Of the 45 isolates, 13 (28.9%) and 14 (31.1%) were resistant to tetracycline and ciprofloxacin, respectively, and 3 (6.7%) showed reduced susceptibility to cefixime (although the susceptibility category was not determined). These results indicate that several isolates of N. subflava have acquired resistance or intermediate resistance to various antimicrobial agents, including penicillin, cephalosporin, tetracycline, and ciprofloxacin. The present study may thus confirm that N. subflava may be involved in the emergence of N. gonorrhoeae strains with either intermediate or total resistance to penicillin or cephalosporin by the horizontal genetic exchange of the penA gene.

In vitro synergistic effects of double combinations of beta-lactams and azithromycin against clinical isolates of Neisseria gonorrhoeae.

In Japan, Neisseria gonorrhoeae, a sexually transmitted pathogen, has recently shown significant resistance to various antimicrobial agents. In this study, a checkerboard method was utilized to investigate the in vitro activities of cefixime (CFIX), cefteram (CFTM), or amoxicillin (AMPC) in combination with azithromycin (AZM) against 25 clinical isolates of N. gonorrhoeae. Synergy, defined as a fractional inhibitory concentration (FIC) index of less than or equal to 0.50, was observed in 32% of isolates with CFIX+AZM, 12% of isolates with CFTM+AZM, and 4% of isolates with AMPC+AZM. Moreover, partial synergy, defined as an FIC index of greater than 0.50 and less than 1, was observed in 44% of isolates with CFIX+AZM, 68% of isolates with CFTM+AZM, and 52% of isolates with AMPC+AZM. In particular, as a result of the combination of CFIX and AZM, for all isolates, significant reductions were observed in the median CFIX minimum inhibitory concentration (MIC; from 0.25 to 0.008 microg/ml; P < 0.0001) and the median AZM MIC (from 0.12 to 0.03 microg/ml; P < 0.0001). However, antagonism, defined as an FIC index of greater than 1, was observed in only 4% of the isolates with both CFIX+AZM and CFTM+AZM, while it was seen in 12% of the isolates with AMPC+AZM. To our knowledge, this is the first study to demonstrate that the in vitro activity of CFIX against N. gonorrhoeae can be significantly enhanced in combination with AZM.

[A case report of complication liked jejunal conduit syndrome induced by reconstruction of ileal conduit].

The patient was a 77-year-old man who underwent radical cystectomy and ileal conduit urinary diversion due to bladder cancer in 1989. A stenosis of the right uretero-ileal anastomosis occurred in 1992, and of the left uretero-ileal anastomosis in 1999. These were treated with indwelling of a ureteral stent and percutaneous nephrostomy, respectively. He was admitted to our hospital for progressive renal dysfunction due to frequent pyelonephritis. We performed a reconstruction of the ileal conduit urinary diversion and after the removal of the bilateral ureteral stent he complained of nausea and general malaise. The laboratory data showed hyponatremia, hyperkalemia and azotemia, which were diagnosed as complication liked jejunal conduit syndrome. He was treated with hydration and salt supplementation. With regard to this case, we considered that a long ileal conduit close to the jejunum and renal dysfunction caused the complication liked jejunal conduit syndrome. Careful observation and follow-up laboratory examination should be performed if the patient has renal dysfunction and a long conduit near the jejunum is used for the ileal conduit.