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ABSTRACT: Acute bladder distension causes various morphological and functional changes, in part through altered gene expression. We aimed to investigate the physiological role of PTHrP, which is upregulated in an acute bladder distension model in female rats. In the control Empty group, bladders were kept empty for 6 h, and in the Distension group, bladders were kept distended for 3 h after an artificial storing-voiding cycle for 3 h. In the Distention group bladder, upregulation of transcripts was noted for three genes reported to be upregulated by stretch in the cultured bladder smooth muscle cells (BSMC) in vitro. Further transcriptome analysis by microarray identified PTHrP as the 2second highest gene upregulated in Distension group bladder, among more than 27,000 genes. Localization of PTHrP and its functional receptor, PTH/PTHrP receptor 1(PTH1R), were analyzed in the untreated rat bladders and cultured bladder cells using real-time RT-PCR and immunoblotting, which revealed that PTH1R and PTHrP were more predominantly expressed in smooth muscle than in urothelium. Exogenous PTHrP peptide (1-34) increased intracellular cAMP level in cultured BSMC. In organ bath study using bladder strips, the PTHrP peptide caused a marked reduction in the amplitude of spontaneous contraction, but caused only modest suppression for carbachol-induced contraction. In in vivo functional study by cystometrogram, the PTHrP peptide decreased voiding pressure and increased bladder compliance. Thus, PTHrP is a potent endogenous relaxant of bladder contraction, and autocrine or paracrine mechanism of PTHrP-PTH1R axis is a physiologically relevant pathway functioning in the bladder.
Endocrinology 04/2013; · 4.46 Impact Factor
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ABSTRACT: Rev-erbα, a component of the circadian clock, has also been known as a nuclear receptor that lacks activation function domain 2, functioning as a ligand-dependent transcriptional repressor. However, we recently reported that Rev-erbα activates connexin43 transcription by forming a complex with Sp1. Here we show that heme, a REV-ERB ligand, is dispensable for this novel mechanism and that Rev-erbβ, having homologies with Rev-erbα, does not activate connexin43, but competes with the Rev-erbα/Sp1. The A/B region of Rev-erbα, which is not conserved in Rev-erbβ, is a crucial activating domain, while the ligand binding domain, conserved in Rev-erbβ, functions as a competitor.
FEBS letters 11/2012; · 3.54 Impact Factor
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Hiroko Kimura,
Tomomi Kamba,
Takeshi Atsuta,
Yuki Makino,
Masashi Oae,
Yoshiyuki Matsui, Masaaki Imamura,
Yousuke Shimizu,
Takahiro Inoue,
Kazutoshi Okubo,
Koji Yoshimura,
Akihiro Kanematsu,
Hiroyuki Nishiyama,
Osamu Ogawa
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ABSTRACT: A 49-year-old man presented with elevated serum creatinine. He had histories of pancreatic pseudocyst caused by alcoholic pancreatitis and right nephrectomy due to renal cell carcinoma. Computed tomographic scan demonstrated that a subcapsular renal cyst appeared adjacent to the pancreatic pseudocyst and compressed the parenchyma of the left kidney. Since the renal subcapsular cyst was suspected to directly communicate with the pancreatic pseudocyst, endoscopic ultrasound-guided transgastric drainage of pancreatic pseudocyst was performed. After the intervention, the renal subcapsular cyst disappeared without formation of a pancreatic fistula and renal dysfunction was promptly improved. He remains free of relapse for more than 1 year.
Hinyokika kiyo. Acta urologica Japonica 08/2012; 58(8):421-4.
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Hiromitsu Negoro,
Akihiro Kanematsu,
Masao Doi,
Sylvia O Suadicani,
Masahiro Matsuo, Masaaki Imamura,
Takeshi Okinami,
Nobuyuki Nishikawa,
Tomonori Oura,
Shigeyuki Matsui,
Kazuyuki Seo,
Motomi Tainaka,
Shoichi Urabe,
Emi Kiyokage,
Takeshi Todo,
Hitoshi Okamura,
Yasuhiko Tabata,
Osamu Ogawa
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ABSTRACT: Nocturnal enuresis in children and nocturia in the elderly are two highly prevalent clinical conditions characterized by a mismatch between urine production rate in the kidneys and storage in the urinary bladder during the sleep phase. Here we demonstrate, using a novel method for automated recording of mouse micturition, that connexin43, a bladder gap junction protein, is a negative regulator of functional bladder capacity. Bladder connexin43 levels and functional capacity show circadian oscillations in wild-type mice, but such rhythms are completely lost in Cry-null mice having a dysfunctional biological clock. Bladder muscle cells have an internal clock, and show oscillations of connexin43 and gap junction function. A clock regulator, Rev-erbα, upregulates connexin43 transcription as a cofactor of Sp1, using Sp1 cis-elements of the promoter. Therefore, circadian oscillation of connexin43 is associated with the biological clock and contributes to diurnal changes in bladder capacity, which avoids disturbance of sleep by micturition.
Nature Communications 01/2012; 3:809. · 7.40 Impact Factor
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Akihiro Kanematsu,
Shiro Tanaka,
Kazuyoshi Johnin,
Shina Kawai,
Shigeru Nakamura, Masaaki Imamura,
Koji Yoshimura,
Yoshihide Higuchi,
Shingo Yamamoto,
Yusaku Okada,
Hideo Nakai,
Osamu Ogawa
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ABSTRACT: OBJECTIVE: We created software for patterning uroflowmetry (UFM) curves, and validated its utility. PATIENTS AND METHODS: The software patterns a given UFM curve upon four parameters: sex, voided volume, maximal flow rate, and amplitude of fluctuation. Using the software, 6 urologists from 4 institutes assessed 30 test curves. Further, 329 UFM curves obtained from children presenting to 3 institutes for daytime and/or nighttime wetting were assessed. Clinical presentation was divided into 3 groups: group A, daytime incontinence; group B, non-monosymptomatic nocturnal enuresis without daytime wetting; and group C, monosymptomatic nocturnal enuresis. RESULTS: Using the software, inter-rater agreement ranged from 0.85 to 1.00 (mean, 0.93 ± 0.04). It could pattern 310 out of 329 clinical curves. In each institute, the tower pattern was prevalent according to severity of daytime symptoms, although not significantly. The merged data showed that the percent tower pattern significantly correlated with presence of daytime symptoms (groups A, B, and C, 29.7%, 27.0%, and 16.3%, respectively; p < 0.05). No correlation with daytime symptoms was noted for fluctuated (staccato and interrupted) and plateau patterns. CONCLUSION: The software creates a common platform for evaluating pediatric UFM, enabling extraction of common and biased features of different cohorts, and their integration into one single cohort.
Journal of pediatric urology 12/2011; · 1.38 Impact Factor
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ABSTRACT: Basic fibroblast growth factor is a candidate causative factor of detrusor overactivity in bladder outlet obstruction cases through up-regulation of the gap junction protein connexin 43. We addressed the transcriptional and behavioral implications of this axis.
Cx43 and Cx45 mRNA expression was assessed by real-time reverse transcriptase-polymerase chain reaction in the bladder of a rat bladder outlet obstruction model and in cultured rat bladder smooth muscle cells with and without basic fibroblast growth factor treatment. Involvement of the extracellular signal regulated kinase 1/2-activator protein-1 pathway was evaluated by immunofluorescence study and a promoter-reporter assay in bladder smooth muscle cells. The effect of basic fibroblast growth factor on micturition behavior was measured in unrestrained rats under a 12-hour light/dark cycle using a controlled release system from gelatin hydrogels fixed on the bladder. The expression of extracellular signal regulated kinase 1/2 and connexin 43 protein was assessed by Western blotting of rat bladder protein.
Cx43 but not Cx45 mRNA expression was increased in the bladder of the obstruction model and in bladder smooth muscle cells treated with basic fibroblast growth factor. The mitogen-activated and extracellular signal-regulated kinase kinase inhibitor PD98059 blocked the stimulatory effect of basic fibroblast growth factor on connexin 43 protein expression and promoter activity, which was also decreased by mutation or deletion of an activator protein-1 cis-element of the connexin 43 promoter. In vivo application of basic fibroblast growth factor on the bladder increased urinary frequency during the latter half of the dark phase, ie the late active phase of rats (F = 5.1, 2-way ANOVA p <0.05). The expression of phospho-extracellular signal regulated kinase 1/2 and connexin 43 protein was increased in the bladder.
The extracellular signal regulated kinase 1/2-activator protein-1-connexin 43 axis could be a potential therapeutic target for increased urinary frequency.
The Journal of urology 06/2011; 185(6):2398-404. · 4.02 Impact Factor
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Masashi Oae,
Kazutoshi Okubo,
Yuichi Uemura,
Takeshi Atsuta,
Hiroko Kimura,
Yuki Makino,
Yoshiyuki Matsui, Masaaki Imamura,
Yousuke Shimizu,
Takahiro Inoue,
Tomomi Kamba,
Koji Yoshimura,
Akihiro Kanematsu,
Hiroyuki Nishiyama,
Daisuke Kawabata,
Osamu Ogawa
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ABSTRACT: A 66-year-old man presented with multiple bilateral renal nodular lesions demonstrated by enhanced computed tomographic scan. He had a history of autoimmune pancreatitis and renal cell carcinoma, which had been treated with partial nephrectomy. We performed renal biopsy under ultrasound guidance. Pathological examination revealed plasma cell infiltration to the renal interstitium. Serum IgG4 level was high and we diagnosed as IgG4-related tubulointerstitial nephritis. After one month of oral steroid therapy the multiple nodular lesions disappeared.
Hinyokika kiyo. Acta urologica Japonica 06/2011; 57(6):309-13.
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ABSTRACT: A 35-day-old male infant was referred to our hospital for bilateral hydroureteronephrosis on postnatal ultrasonography. He was born at 37 weeks and 2 days of gestation, weighing 2,765 g. He was diagnosed with bilateral primary obstructive megaureter. During observation, at 3 months and 2 weeks of age, he presented with dehydration and showed hyponatremia and hyperkalemia. An endocrinological examination led to a diagnosis of pseudohypoaldosteronism type 1 (PHA1). After bilateral percutaneous nephrostomy, PHA1 resolved. He underwent bilateral ureteral tapering and ureteroneocystostomy at 5 months and 3 weeks of age. Since PHA1 recurred after removal of urethral catheter, urethral catheter and bilateral double-J-stent was kept until 8 months old. He was followed-up until 3 years old with no signs of PHA1 recurrence. For an infant with obstructive uropathy or vesicoureteral reflux showing hyponatremia and hyperkalemia, PHA1 should be considered.
Hinyokika kiyo. Acta urologica Japonica 04/2011; 57(4):193-7.
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Masaaki Imamura,
Hiromitsu Negoro,
Akihiro Kanematsu,
Shingo Yamamoto,
Yu Kimura,
Kentaro Nagane,
Toshinari Yamasaki,
Isao Kanatani,
Noriyuki Ito,
Yasuhiko Tabata,
Osamu Ogawa
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ABSTRACT: Overactive bladder is a highly prevalent clinical condition that is often caused by bladder outlet obstruction (BOO). Increased coupling of bladder smooth muscle cells (BSMC) via gap junctions has been hypothesized as a mechanism for myogenic bladder overactivity in BOO, although little is known about the regulatory system underlying such changes. Here, we report the involvement of basic fibroblast growth factor (bFGF) and connexin 43, a bladder gap junction protein, in bladder overactivity. BOO created by urethral constriction in rats resulted in elevated bFGF and connexin 43 levels in the bladder urothelium and muscle layer, respectively, and muscle strips from these bladders were more sensitive than those from sham-operated controls to a cholinergic agonist. In vitro bFGF treatment increased connexin 43 expression in cultured rat BSMC via the ERK 1/2 pathway. This finding was supported by another in vivo model, where bFGF released from gelatin hydrogels fixed on rat bladder walls caused connexin 43 upregulation and gap junction formation in the muscle layer. Bladder muscle strips in this model showed increased sensitivity to a cholinergic agonist that was blocked by inhibition of gap junction function with alpha-glycyrrhetinic acid. Cystometric analyses of this model showed typical features of detrusor overactivity such as significantly increased micturition frequency and decreased bladder capacity. These findings suggest that bFGF from the urothelium could induce bladder hypersensitivity to acetylcholine via gap junction generation in the smooth muscle, thereby contributing to the myogenic overactivity of obstructed bladders.
AJP Renal Physiology 05/2009; 297(1):F46-54. · 4.42 Impact Factor
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ABSTRACT: A metastatic penile tumor of malignant melanoma is very rare. The patient had priapism caused by metastatic penile tumor and the primary cancer was in the end stage. However, in this case, we could relieve the dysuria and pain by percutaneous cystostomy and partial penectomy. The corpus spongiosumcorpus cavernosum shunt could not relieve the priapism and might have accelerated the metastasis.
Hinyokika kiyo. Acta urologica Japonica 04/2009; 55(3):149-52.
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ABSTRACT: The adhesion and proliferation of human adipo-stromal cells was investigated for poly(ethylene terephthalete) (PET) films of two-dimensional (2D) substrate and non-woven fabrics of three-dimensional (3D) substrate after seeding at the same cell density and culturing at the same medium volume to surface area of the substrates ratio. When seeded by a static seeding method, more cells adhered on the film than on the non-woven fabric. However, the number ratio of cells proliferated to those initially adhered was similar. For the non-woven fabric, cell proliferation was enhanced by an agitated culture method. NaOH treatment introduced carboxyl groups into the surface of substrates, irrespective of the substrate type. Cell adhesion of a peptide (Arg-Gly-Asp, RGD) was chemically immobilized through the carboxyl groups on the non-woven fabric surface at a density of 10 pmol/cm2. RGD immobilization significantly increased the number of cells adhered after the agitated seeding method, but did not affect the cell proliferation. Phosphorylation of focal adhesion kinase (FAK) was also enhanced by the RGD immobilization on the PET non-woven fabrics.
Journal of Biomaterials Science Polymer Edition 02/2009; 20(5-6):721-36. · 1.69 Impact Factor
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ABSTRACT: Human adipo-stromal cells were incubated in culture medium containing basic fibroblast growth factor (bFGF) or in a cell-culture dish coated with bFGF. The effect of the two existence modes of bFGF on cell attachment and differentiation was evaluated. The morphology and number of cells were similar for both the bFGF modes. However, the extracellular signal-regulated kinase 1 and 2 (ERK1/2) activation of cells incubated with bFGF in the medium was stronger and lasted for a longer time period than that of bFGF in the coated form. When adipo-stromal cells were incubated in an adipogenic differentiation medium, irrespective of the mode of bFGF added, the mRNA expression of peroxisome proliferator-activated receptor gamma2 (PPRAgamma2) and fatty acid binding protein 2 (aP2), the glycerol-3-phosphate dehydrogenase (GPDH) activity and the cell accumulation of oil lipids were all suppressed. bFGF in the solution form inhibited the PPARgamma2 expression to a high extent compared with that in the coated form. It is possible that bFGF-induced ERK1/2 activation consequently inhibited the adipogenic differentiation of adipo-stromal cells.
Journal of Biomaterials Science Polymer Edition 02/2009; 20(4):483-94. · 1.69 Impact Factor
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ABSTRACT: The objective of this study is to investigate the spreading area, proliferation and adipogenic differentiation of adipo-stromal cells cultured on the surface of self-assembled monolayers (SAM) prepared by alkanethiols with hydroxyl (OH), methyl (CH(3)), amine (NH(2)) and carboxyl terminal groups (COOH) or the mixture at different ratios. A modified enzyme-linked immunosorbent assay (ELISA) examination revealed that a high adsorption of vitronectin and fibronectin was observed for the SAM of NH(2) or the mixed SAM of OH and NH(2) groups and the mixed SAM of OH and CH(3) or OH and COOH groups, respectively. The cell spreading area and the proliferation level were higher for the SAM of NH(2) or COOH or the mixed SAM of OH and NH(2) or OH and COOH groups than those of other substrates. When incubated in an adipogenic differentiation medium, the cells showed a high level of glycerol-3-phosphate dehydrogenase (GPDH) activity for the SAM of CH(3) or the mixed SAM of OH and COOH groups. In addition, a high mRNA expression of peroxisome proliferator-activated receptor gamma2 (PPRAgamma2) and fatty acid binding protein 2 (aP2) was observed. For the SAM of NH(2) or COOH groups, the strong activation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) was observed while the mRNA expression of connective tissue growth factor (CTGF) and cysteinrich 61 (CYR61) was enhanced. The proliferation of the cells was significantly suppressed by adding an inhibitor of ERK1/2. The mRNA expression of PPAR gamma2 was significantly induced by adding the inhibitor. It is concluded that the proliferation and adipogenic differentiation of adipo-stromal cells depended on the chemical composition of substrate surface, although the extent was influenced by that of ERK1/2 activation.
Journal of Biomaterials Science Polymer Edition 02/2008; 19(7):893-914. · 1.69 Impact Factor
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ABSTRACT: An ideal biomaterial for urethral reconstruction has not been developed. To create a urethral graft biomaterial with optimal biodegradability and biocompatibility, a copoly(L-lactide/epsilon-caprolactone) [P(LA/CL)] fabric tube was combined with a type I collagen sponge. The P(LA/CL) fibers were knitted into a vascular stent style (Type 1) or weaved into a mesh style (Type 2) to prepare P(LA/CL) tubes. The tubes were dipped in aqueous collagen solution and lyophilyzed to prepare the P(LA/CL)-collagen sponge graft. The grafts were applied to a 1.5-cm rabbit urethral defect (n = 14 for each condition), and tissue repair was evaluated using urethrographical, urethroscopical, and histological examination 1, 3, and 6 months after surgery. Although epithelialization was observed after 1 month in all Type 1 grafts, stenoses, fistulae, or stone formation was seen in 7 of the rabbits. In some cases, P(LA/CL) fibers prolapsed into the urethral lumen, causing stone formation. Only 3 rabbits survived for 6 months, and 2 of these had stenoses. For the Type 2 graft, all urethras were patent, without fistulae or stenoses, over the entire observation period. Histologically, urethral structure was disorganized for the Type 1 graft, whereas the urethral tissue on the Type 2 graft was slightly fibrotic but completely epithelialized and supported by a regenerated smooth muscle layer at 6 months. These findings suggest that creation of a scaffold suitable for urethral tissue regeneration will depend not only on the biomaterial composition, but also on the fabrication technique.
Tissue Engineering 01/2008; 13(12):2933-40. · 4.02 Impact Factor
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ABSTRACT: Bladder hypertrophy is a general consequence of bladder outlet obstruction (BOO) and a typical phenomenon observed in clinical urologic diseases such as benign prostatic hyperplasia and neurogenic bladder. It is characterized by smooth muscle hyperplasia, altered extracellular matrix composition, and increased contractile function. Various growth factors are likely involved in hypertrophic pathophysiology, but their functions remain unknown. In this report, the role of basic fibroblast growth factor (bFGF) was investigated using a rat bladder smooth muscle cell (BSMC) culture system and an original animal model, in which bFGF was released from a gelatin hydrogel directly onto rat bladders. bFGF treatment promoted BSMC proliferation both in vitro and in vivo. In vitro, bFGF downregulated the expression of type I collagen, but upregulated type III collagen. ERK1/2, but not p38MAPK, was activated by bFGF, whereas inhibition of ERK1/2 by PD98059 reversed bFGF-induced BSMC proliferation, type I collagen downregulation, and type III collagen upregulation. In the in vivo release model, bFGF upregulated type III collagen and increased the contractile force of treated bladders. In parallel with these findings, hypertrophied rat bladders created by urethral constriction showed increased urothelial bFGF expression, BSMC proliferation, and increased type III collagen expression compared with sham-operated rats. These data suggest that bFGF from the urothelium could act as a paracrine signal that stimulates the proliferation and matrix production of BSMC, thereby contributing to the hypertrophic remodeling of the smooth muscle layer.
American journal of physiology. Renal physiology 11/2007; 293(4):F1007-17. · 3.68 Impact Factor
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Isao Kanatani,
Tomonori Ikai,
Arimichi Okazaki,
Jun-Ichiro Jo,
Masaya Yamamoto, Masaaki Imamura,
Akihiro Kanematsu,
Shingo Yamamoto,
Noriyuki Ito,
Osamu Ogawa,
Yasuhiko Tabata
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ABSTRACT: The feasibility and mechanism of gene delivery by pullulan-spermine, a recently developed cationic polysaccharide, were investigated. Pullulan-spermine-mediated transfection of plasmid DNA resulted in greatly reduced cytotoxicity and a 10-fold increase in the level of gene expression when compared to Lipofectamine 2000, a commercially available cationic lipid. Additionally, after transfection of p53-expressing plasmid DNA by pullulan-spermine but not Lipofectamine 2000, the in vitro proliferation of T24 cells was significantly reduced. Pullulan-spermine-mediated gene expression was inhibited by both chlorpromazine of clathrin-mediated endocytosis inhibitor and methyl-beta-cyclodextrin and filipin of raft/caveolae inhibitors. We conclude that pullulan-spermine is a promising carrier for gene transfection, and that cellular uptake of pullulan-spermine-plasmid DNA complexes is mediated by clathrin- and raft/caveolae-dependent endocytotic pathways.
Journal of Controlled Release 12/2006; 116(1):75-82. · 5.73 Impact Factor
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ABSTRACT: Tissue regeneration on acellular matrix grafts has great potential for therapeutic organ reconstruction. However, hollow organs such as the bladder require smooth muscle cell regeneration, the mechanisms of which are not well defined. We investigated the mechanisms by which bone marrow cells participate in smooth muscle formation during urinary bladder regeneration, using in vivo and in vitro model systems. In vivo bone marrow cells expressing green fluorescent protein were transplanted into lethally irradiated rats. Eight weeks following transplantation, bladder domes of the rats were replaced with bladder acellular matrix grafts. Two weeks after operation transplanted marrow cells repopulated the graft, as evidenced by detection of fluorescent staining. By 12 weeks they reconstituted the smooth muscle layer, with native smooth muscle cells (SMC) infiltrating the graft. In vitro, the differential effects of distinct growth factor environments created by either bladder urothelial cells or bladder SMC on phenotypic changes of marrow cells were examined. First, supernatants of cultured bladder cells were used as conditioned media for marrow cells. Second, these conditions were reconstituted with exogenous growth factors. In each case, a growth factor milieu characteristic of SMC induced an SMC-like phenotype in marrow cells, whereas that of urothelial cells failed. These findings suggest that marrow cells differentiate into smooth muscle on acellular matrix grafts in response to the environment created by SMC.
American Journal Of Pathology 03/2005; 166(2):565-73. · 4.89 Impact Factor
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ABSTRACT: A 55-year-old woman had complained of lower abdominal pain for 5 years. A computed tomographic CT scan and magnetic resonance imaging revealed a heterogeneous density mass on the left side of the bladder. CT-guided needle biopsy showed only inflammatory cell infiltration without histological diagnosis. To confirm the diagnosis, the mass was resected together with part of the bladder and peritoneum. On the cut surface, the mass was centrally pale yellow and adherent to the thickened hard wall of the bladder and peritoneum. Pathological diagnosis was chronic granulomatous inflammation with actinomycosis. This is, to our knowledge, the 12th case report of vesical actinomycosis in Japan. Although actinomycosis is very rare in urological field, abdominal actinomycosis should be considered as the differential diagnosis of any inflammatory abdominal mass.
Hinyokika kiyo. Acta urologica Japonica 12/2003; 49(11):659-61.
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ABSTRACT: We report a case of adult sacrococcygeal teratoma resected by an abdominosacral approach. A cystic mass 13 cm in diameter in the pelvic cavity and left hydronephrosis were detected incidentally by abdominal computed tomographic (CT) scan in a 55-year-old man. The pelvic tumor extending from the presacral area to the coccyx was resected via a combined abdominal and transsacral approach. The resected specimen weighed 700 g and the pathological diagnosis was mature teratoma. While the sacrococcygeal area is the most frequent site of teratoma in infants, it is a rare site in adults. This is to our knowledge, the 30th case report of adult sacrococcygeal teratoma in Japan. At one month after the operation the patient had no bowel dysfunction and no dysbasia, but he had mild bladder dysfunction requiring self-catheterization twice a day at twelve months. The patient had no evidence of disease at twenty months after the operation.
Hinyokika kiyo. Acta urologica Japonica 11/2002; 48(10):599-601.
