Jun Kamei

Preventive Effects of Long-Term Caloric Restriction on Aging Related In Vivo Bladder Dysfunction and Molecular Biological Changes in the Bladder and Dorsal Root Ganglia in Rats

PURPOSE We evaluated aging related bladder dysfunctions and biological changes in the bladder and dorsal root ganglia in rats. We also investigated whether long-term caloric restriction may have preventive effects on these changes. MATERIALS AND METHODS Male Fischer 344 rats were divided into a young group (age 6 months) and an old group (age 25 to 28 months), each with free access to normal food, and an old group (age 25 to 28 months) with food restricted to 3 days per week. Conscious cystometry, cDNA microarray analysis, immunohistochemistry and oxidative stress measurements of the bladder and dorsal root ganglia were performed. RESULTS The old group with free access to normal food showed higher threshold pressure, more nonvoiding contractions and lower bladder compliance than the young group with free access to food. Old rats with free access showed greater post-void residual volume and lower voiding efficiency than old rats with caloric restriction and young rats. In the old group with free access 83 genes in the bladder and 48 in the L6 dorsal root ganglia were up-regulated compared with old rats with caloric restriction and young rats. These genes were mostly related to immune and inflammatory responses. Immunohistochemistry showed stronger expression of the immune response protease Gzm (granzyme) B and the collagenase Mmp13 (matrix metalloproteinase-13) in the bladder of old rats with free access vs old rats with caloric restriction and young rats. The level of malondialdehyde, an oxidative stress marker, was higher in the bladder of old rats with free access than in young rats but there was no difference between old rats with caloric restriction and young rats with free access to food. CONCLUSIONS In rats aging leads to storage and voiding dysfunctions associated with immune and inflammatory related responses in the bladder and dorsal root ganglia, and with increased oxidative stress in the bladder. Caloric restriction reduced these aging related changes.

Toll-like receptor 7 is overexpressed in the bladder of Hunner-type interstitial cystitis, and its activation in the mouse bladder can induce cystitis and bladder pain

Abstract Toll-like receptor 7 (TLR7) is associated with the pathophysiology of systemic lupus erythematosus and Sjögren syndrome, well-known diseases accompanying interstitial cystitis (IC). We studied TLR7 expression in the bladder of patients with Hunner-type IC (HIC) and its functional roles in bladder inflammation and nociception using mice. Bladder biopsy specimens were obtained from patients with HIC. Specimens from the noncancerous portion of the bladder of patients with bladder cancer served as controls. The specimens were examined by immunohistochemistry and real-time polymerase chain reaction of TLR7. Loxoribine (LX), a TLR7 agonist, was instilled in the bladder of C57BL/6N female mice, and TLR7-mRNA expression and histological changes of the bladder, bladder pain–like licking behavior, voiding behavior, cystometry, and bladder afferent nerve activities were investigated. The effects of hydroxychloroquine, a TLR7 antagonist, on the LX-induced changes on cystometry and voiding behavior were studied. The number of TLR7 immuno-reactive cells and the mRNA expression of TLR7 were significantly increased in HIC specimens. Intravesical instillation of LX induced edema, congestion, inflammation, and significantly increased TLR7-mRNA expression in the mouse bladder. Loxoribine-instillation also significantly increased licking behavior, voiding frequency, and afferent nerve activities associated with decreased single-voided volume and intercontraction interval of micturitions. Hydroxychloroquine reversed the LX-induced cystometric and voiding behavioral changes. Toll-like receptor 7 was up-regulated in the bladder mucosa of patients with HIC, and activation of TLR7 in the mouse bladder induced cystitis with sensory hyperactivity of the bladder. Blocking the TLR7 pathway may be an innovative treatment target of HIC.

Prevalence and characteristics of fecal antimicrobial-resistant Escherichia coli in a cohort of Japanese men undergoing prostate biopsy

To examine resistant Escherichia coli in rectal swab culture of Japanese men undergoing prostate biopsy, and to determine its prevalence, genotypic characteristics and carriage risk factors.

Video‐urodynamic effects of mirabegron, a β3‐adrenoceptor agonist, in patients with low‐compliance bladder

To investigate video‐urodynamic effects of mirabegron, a β3‐adrenoceptor agonist, on low‐compliance bladder.

Diagnostic value of urinary CXCL10 as a biomarker for predicting Hunner type interstitial cystitis

To investigate the feasibility of chemokines and cytokines potentially elevated in the bladder tissue of Hunner type interstitial cystitis (HIC) as urinary markers for distinguishing HIC from non‐Hunner type interstitial cystitis (NHIC)

Age-related changes in function and gene expression of the male and female mouse bladder.

We investigated age-related changes in in vivo and in vitro functions and gene expression of the bladder of male and female mice. Mature and aged (12 and 27–30 month old) C57BL/6 mice of both sexes were used. Frequency volume, conscious free-moving cystometry and detrusor contractile and relaxant properties in in vitro organ bath were evaluated. mRNA expression level of muscarinic, purinergic, and β-adrenergic receptors and gene expression changes by cDNA microarray analysis of the bladder were determined. Cystometry demonstrated storage and voiding dysfunctions with ageing in both sexes. Detrusor strips from aged mice showed weaker contractile responses particularly in the cholinergic component and weaker relaxant responses to isoproterenol. These age-related impairments were generally severer in males. mRNA expression of bladder tissue was decreased for M3 muscarinic receptors in aged males and β2-adrenoceptors in aged females. cDNA microarray analysis results, albeit substantial sex difference, indicated “cell-to-cell signaling and interaction” as the most common feature of age-related gene expression. In summary, aged mice demonstrated voiding and storage dysfunctions resembling to detrusor hyperactivity with impaired contractility (DHIC), which were more pronounced in males. Genomic changes associated with aging may contribute to the age-related bladder functional deterioration in mice.

Risk factors for surgical site infection after transvaginal mesh placement in a nationwide Japanese cohort

We conducted a nationwide survey on perioperative management and antimicrobial prophylaxis of transvaginal mesh surgeries for pelvic organ prolapse in Japan to understand the practice and risk factors for surgical site infection (SSI).

Attenuated lipopolysaccharide-induced inflammatory bladder hypersensitivity in mice deficient of transient receptor potential ankilin1

Transient receptor potential ankyrin 1 (TRPA1) channel expressed by urothelial cells and bladder sensory nerve fibers might act as a bladder mechanosensor and nociceptive transducer. To disclose the role of TRPA1 in bladder function and inflammation-associated hypersensitivity, we evaluated in vitro and in vivo bladder function and inflammatory mechanosensory and nociceptive responses to intravesical lipopolysaccharide (LPS)-instillation in wild type (WT) and TRPA1-knock out (KO) mice. At baseline before treatment, no significant differences were observed in frequency volume variables, in vitro detrusor contractility, and cystometric parameters between the two groups in either sex. LPS-instillation significantly increased voiding frequency and decreased mean voided volume at 24–48 hours after instillation in WT but not in TRPA1-KO mice. LPS-instillation also significantly increased the number of pain-like behavior at 24 hours after instillation in WT mice, but not in TRPA1-KO mice. Cystometry 24 hours after LPS-instillation revealed shorter inter-contraction intervals in the WT mice compared with TRPA1-KO mice. In contrast, inflammatory cell infiltration in the bladder suburothelial layer was not significantly different between the two groups. These results indicate that TRPA1 channels are involved in bladder mechanosensory and nociceptive hypersensitivity accompanied with inflammation but not in physiological bladder function or development of bladder inflammation.

Regional clinical practice variation in urology: Usage example of the Open Data of the National Database of Health Insurance Claims and Specific Health Checkups of Japan

DOI: 10.1111/iju.13840 The Ministry of Health, Labor and Welfare released the second NDB Open Data Japan in October 2017. NDB Open Data Japan is a fundamental table summarizing Japanese national universal healthcare insurance claims. The database comprises almost all (≥95%) claims data regarding medical and dental treatments, and specific health checkups, and it provides us a complete picture of the real-world clinical practice in Japan. The data are openly available for free on the website, http://www.mhlw.go.jp/stf/seisakunitsuite/bunya/0000177182.html. The period in the first and second NDB Open Data Japan were the fiscal year 2014 and 2015 (12 months from April to the next March). Because of its ultimately high representativeness, it enables us to know how widely our urological clinical practice varies by geographic regions. This visualization would wield a notable impact on both physicians and patients to understand the current standard of care, to examine future health policies locally and to promote urological healthcare uniformity globally. Because the spreadsheets of the NDB Open Data were written in insurance technical codes, we believe a physician-friendly reformat would be widely welcomed to enhance its recognition and effective utilization. For example, Table 1 is a list of the prevalence of minimally invasive surgery in nephrectomy and nephroureterectomy for malignancy in each prefecture in the fiscal year 2015, with the prevalence proportion appearing in descending order. Minimally invasive surgery is defined as laparoscopic and minimal incision endoscopic surgery. Robot-assisted surgery was not included due to non-coverage in national universal healthcare insurance at that time. Although minimally invasive surgery accounted for 64.9% throughout Japan, the prefectural proportion widely varied from 93.9% in Miyazaki to 25.0% in Yamagata. Because the share of minimally invasive surgery between 2007 and 2010 was reported to be 42.1% based on the Diagnosis Procedure Combination database, it had increased by half from 2010 to 2015. Regarding geographic regional variation, the rate of minimally invasive surgery was >70% in the western areas, and <60% in the eastern area, except Hokkaido. The previous Diagnosis Procedure Combination database study also pointed out a distribution discrepancy between eastern and western areas. A definitive reason for the variation could not be drawn from the database analysis; however, it would be reasonable to mention that highlyskilled laparoscopic surgeons would likely be produced in high-prevalence areas where there was suitable laparoscopic training, and the increasing number of skilled surgeons contributes further to spreading minimally invasive surgery in the region. A detailed table on prefectural distribution of urological surgical procedures in a physicianfriendly format is provided as Table S1. Some comments should be added for interpretation of the table. First, any individual cell counts <10 were not available to maintain confidentiality. When such a cell was single, all the other cell counts become unavailable to block back calculation. This cell size suppression policy was set to avoid the potential identification risk of individuals. Second, the second NDB Open Data Japan in the fiscal year 2015 separated inpatient and outpatient procedures, whereas the first NDB Open Data Japan in the fiscal year 2014 showed combined data only. Owing to the rules, prefectural counts in extracorporeal shockwave lithotripsy at outpatient settings were unavailable in the fiscal year 2015. Therefore, prefectural inpatient–outpatient combined counts in the fiscal year 2015 were estimated by being prorated from the counterpart of the fiscal year 2014, and then the outpatient counts in the fiscal year 2015 were calculated by combined counts minus inpatient counts. Third, because some patients travel across the prefectures, the hospital-based counts in this manuscript could lead to overor underestimation of residence-based counts. The NDB Open Data included other types of data, such as distribution of age and sex regarding medical procedures, counts of most frequently prescribed drugs for each therapeutic category, and results of specific health checkups. A database with near-complete capture of healthcare services like the NDB has a great potential to solve a wide range of clinical and healthcare questions. In the USA, 100% of the Medicare dataset was used for several analyses including epidemiology study; treatment utilization variation; the relationship among cost, safety, effectiveness and quality; and the Urological Notes

MP8-06 PREVENTIVE EFFECTS OF CALORIC RESTRICTION ON AGING-ASSOCIATED BIOLOGICAL AND MOLECULAR CHANGES IN THE RAT BLADDER AND DORSAL ROOT GANGLIA

maintained on regular diet for 30 weeks. Animals were subjected to the assessment of body weight (BW), body length (BL), waist circumference (WC), body mass index (BMI), blood glucose (BG), plasma insulin (INS), plasma leptin (LEP), total cholesterol (CHO), free fatty acid (FFA) and evaluated for urinary voiding functions. Total body fat measurement, prostate and bladder volumes were analyzed by MRI followed by histological evaluation of the organs. These parameters were used to examine the associations between MS and LUTS. RESULTS: Obesity parameters such as BW, WC, and BMI were significantly higher in B6.V-Lepob/J mice compared to C57BL/6N mice (p<0.01). Higher levels of total CHO and FFA were noted in B6.V-Lepob/ J mice than C57BL/6N mice (p<0.05). These results were concurrent with frequency, lower average urine volume and other urinary voiding dysfunctions inB6.V-Lepob/Jmice.MRIassessmentdemonstratemarked increase in body fat and prostate volume in these mice. Histology of prostate in B6.V-Lepob/J mice showed increased gland crowding and infiltration of immune cells in the stroma, compared to C57BL/6N mice. The regression and correlation analysis indicate that peritoneal fat (R1⁄40.831; p<0.002), BG (R1⁄40.712; p<0.01) and prostate volume (R1⁄40.706; p<0.05) strongly correlate with LUTS whereas BMI, WC, BMI, INS, CHO and FFA moderately correlate with the prevalence of voiding dysfunctions. CONCLUSIONS: Our findings suggests that LUTS may be attributable in part to obesity and MS. Further examination of our in vivo model may lead to understand the underlying pathophysiological mechanisms of obesity-induced LUTS in humans.