Safety Assessment of Ultrasound-Assisted Intravesical Chemotherapy in Normal Dogs: A Pilot Study.

Intravesical chemotherapy after transurethral resection is a treatment option in patients with non-muscle invasive bladder cancer. The efficacy of intravesical chemotherapy is determined by the cellular uptake of intravesical drugs. Therefore, drug delivery technologies in the urinary bladder are promising tools for enhancing the efficacy of intravesical chemotherapy. Ultrasound-triggered microbubble cavitation may enhance the permeability of the urothelium, and thus may have potential as a drug delivery technology in the urinary bladder. Meanwhile, the enhanced permeability may increase systemic absorption of intravesical drugs, which may increase the adverse effects of the drug. The aim of this preliminary safety study was to assess the systemic absorption of an intravesical drug that was delivered by ultrasound-triggered microbubble cavitation in the urinary bladder of normal dogs. Pirarubicin, a derivative of doxorubicin, and an ultrasound contrast agent (Sonazoid) microbubbles were administered in the urinary bladder. Ultrasound (transmitting frequency 5 MHz; pulse duration 0.44 µsec; pulse repetition frequency 7.7 kHz; peak negative pressure -1.2 MPa) was exposed to the bladder using a diagnostic ultrasound probe (PLT-704SBT). The combination of ultrasound and microbubbles did not increase the plasma concentration of intravesical pirarubicin. In addition, hematoxylin and eosin staining showed that the combination of ultrasound and microbubble did not cause observable damages to the urothelium. Tissue pirarubicin concentration in the sonicated region was higher than that of the non-sonicated region in two of three dogs. The results of this pilot study demonstrate the safety of the combination of intravesical pirarubicin and ultrasound-triggered microbubble cavitation, that is, ultrasound-assisted intravesical chemotherapy.

Clinical characteristics in urolithiasis formation according to body mass index.

Although urolithiasis is considered to be strongly associated with lifestyle habits, there are numerous cases in which urolithiasis develops despite a non-obese body type or healthy lifestyle habits. However, in clinical practice, the diet therapy and lifestyle changes instructed for the prevention of recurrence of urolithiasis are almost identical in numerous cases. Therefore, the present study examined the effect of body mass index (BMI) on urolithiasis and its surrounding environment in patients, by analyzing the number of normal- and high-BMI (healthy and overweight) patients with urolithiasis. The present study analyzed 63 patients with urolithiasis for whom height and weight were measured at our hospital (Tokyo Medical University Ibaraki Medical Center). BMI <25 represents healthy body types and BMI ≥25 BMI defines overweight body types. Thus, patients were then grouped by BMI-defined body type using a threshold value of 25 accordingly. It was observed that a higher percentage of males were obese compared with females. Upon comparing the normal- and high-BMI groups, no significant difference was observed in uric acid level, urine pH or calculus number between the two groups. Liver computed tomography (CT) values were significantly lower in the high-BMI group compared with the normal-BMI group. There was no significant correlation between calculus size counts and BMI. However, a significant negative correlation was observed between BMI and the liver CT value. These results suggest that liver CT values correlated negatively with BMI, but the data indicates that other mechanisms unassociated with a fatty liver may be involved in urolithiasis in non-obese patients. The results of the present study suggest that physicians should consider the mechanism involved in preventing the recurrence of urolithiasis.

Giant retroperitoneal schwannoma in a 52-year-old man.

Schwannoma in the retroperitoneal space is rare, and it is extremely rare in patients with no history of neurofibromatosis. We present a case of giant retroperitoneal schwannoma in a 52-year-old man who did not have neurofibromatosis. Because malignant transformation would be extremely rare in this circumstance, close imaging follow-up could avert the necessity for complete resection. The possibility of schwannoma should be considered when evaluating retroperitoneal tumors with the characteristic findings, even if there is no connection between the tumor and the intervertebral foramina.

Molecular targeted therapies of renal cell carcinoma considering life stage of the patient: Two case reports.

Molecular targeted therapies have markedly improved the prognosis of metastatic renal cell carcinoma (RCC) and patients are able to receive specific treatments depending on their condition. The cases of two patients who presented with large RCC with very different statuses were examined in the current study. One of the patients was elderly with numerous comorbidities, and the other was young and had Case 1 was an elderly female with various comorbidities including a history of cerebral infarction and impaired eyesight. A 7 cm solid mass in the left kidney, pulmonary metastases and a shrunken right kidney were identified. Conservative treatment was selected and the patient underwent treatment with oral sorafenib for 5 years without experiencing disease progression. Case 2 was a middle-aged male in a good general condition who had a 19 cm left renal mass and multiple metastases. Due to the tumor size, the patient was initially treated with oral pazopanib to reduce the renal tumor volume and was able to undergo left radical nephrectomy after 6 months. Molecular targeted therapies were thus used to treat these patients taking into account each patient's life stage; case 1 was treated without surgery for six years and case 2 received treatment as a neoadjuvant.

Granulomatosis with Polyangiitis (Wegener's Granulomatosis) Accompanied by Dysuria.

A 65-year-old male visited us with complaints of retarded urination, dysuria, gross hematuria, and fever. Urinalysis showed pyuria. Prostatic tumor with lung metastasis was suspected from computed tomography and magnetic resonance imaging. Transurethral prostatic biopsy and bronchoscopic biopsy only revealed fibrinoid necrosis and inflammatory infiltration. Right lateral maxillary sinusitis was also found by MRI. ANCA testing was positive with specificity for anti-PR3 (PR3-ANCA). On the basis of these results, Granulomatosis with polyangiitis (GPA) was diagnosed. GPA involving the prostate gland is unusual, and only a few cases have previously been reported.

Annular rectal constriction caused by infiltrating bladder cancer: A case report.

A 69-year-old man was referred to our hospital with chief complaints of urinary incontinence, pollakiuria, thin stools and edema of the left lower extremity. Computed tomography and magnetic resonance imaging revealed bilateral hydronephrosis, thickening of the rectal and posterolateral bladder walls, and enlargement of the left obturator lymph nodes, suggesting metastasis. The carcinoembryonic antigen and carbohydrate antigen 19-9 levels were 5.6 ng/ml and 460.1 U/ml, respectively. A transurethral bladder biopsy revealed grade 2 urothelial carcinoma with glandular differentiation. A transrectal needle biopsy suggested cancer infiltration of the rectal wall and an annular rectal constriction caused by an infiltrating bladder cancer was diagnosed. Three courses of gemcitabine and cisplatin chemotherapy were administered and demonstrated good efficacy.

Incidental Presacral Myelolipoma Resembling the Liposarcoma: A Case Report and Literature Review.

Presacral myelolipomas are rare, benign, asymptomatic tumors composed of mature adipose tissue and hematopoietic elements, but fewer than 50 cases have been reported in the literature. They are usually discovered incidentally during imaging studies and are often misdiagnosed as liposarcoma, which have a malignant nature, because the imaging findings of myelolipoma can be similar to those of liposarcoma. It is challenging to distinguish presacral myelolipomas from other presacral fat-containing tumors without performing a histological examination. We should consider the possibility of a malignant tumor, and imaging-guided biopsy carries a risk of tumor spread along the biopsy tract. Therefore, surgical management might sometimes be required; however, it is not necessary in all cases. We present an incidentally detected case of presacral myelolipoma that was difficult to differentiate from other malignant tumors in a 71-year-old male.

Development of pathological diagnostics of human kidney cancer by multiple staining using new fluorescent Fluolid dyes.

New fluorescent Fluolid dyes have advantages over others such as stability against heat, dryness, and excess light. Here, we performed simultaneous immunostaining of renal tumors, clear cell renal cell carcinoma (RCC), papillary RCC, chromophobe RCC, acquired cystic disease-associated RCC (ACD-RCC), and renal angiomyolipoma (AML), with primary antibodies against Kank1, cytokeratin 7 (CK7), and CD10, which were detected with secondary antibodies labeled with Fluolid-Orange, Fluolid-Green, and Alexa Fluor 647, respectively. Kank1 was stained in normal renal tubules, papillary RCC, and ACD-RCC, and weakly or negatively in all other tumors. CK7 was positive in normal renal tubules, papillary RCC, and ACD-RCC. In contrast, CD10 was expressed in renal tubules and clear cell RCC, papillary RCC, AML, and AC-RCC, and weakly in chromophobe RCC. These results may contribute to differentiating renal tumors and subtypes of RCCs. We also examined the stability of fluorescence and found that fluorescent images of Fluolid dyes were identical between a tissue section and the same section after it was stored for almost three years at room temperature. This indicates that tissue sections can be stored at room temperature for a relatively long time after they are stained with multiple fluorescent markers, which could open a door for pathological diagnostics.

Decrease rate of the renal diameter in chronic hemodialysis patients.

We here present the results of ultrasonographic (US) evaluations on the alteration of renal diameter of chronic HD patients. Of 109 outpatient HD patients who had neither severe acquired cystic disease of the kidney nor hereditary polycystic kidney disease, we performed US two or three times to measure their maximum renal diameter (mean of both kidneys), and the yearly alteration rate was calculated. The average interval of the two measurements was 35.9 months, and the average HD duration from the HD induction to the first measurement was 29.5 months. The average decrease rate of renal diameter was 4.34 ± 0.4 (SE) mm/year. No statistical difference was seen on the decrease rate in relation to gender, age and original disease (among three groups, glomerulonephritis and IgA nephropathy, diabetes, and others including hypertension). However, the decrease rate was large when the first measurement was close to the induction of hemodialysis, suggesting that the alteration rate reduced according to the hemodialysis vintage (5.3 ± 0.8 mm/year, first measurement not more than 10 months after induction of HD and 1.5 ± 1.6 mm/year, first measurement more than 80 months after induction of HD). Renal diameter decreased approximately 4.3 mm each year, and the decrease rate slowed as the length of time on dialysis increased.

Preoperative prediction of malignant involvement of resected ureters in patients undergoing radical cystectomy for bladder cancer.

OBJECTIVE: To investigate preoperative predictors of ureteral involvement of bladder malignancy and to develop a novel preoperative model for the prediction of ureteral involvement in bladder cancer patients undergoing radical cystectomy. METHODS: This study included 197 consecutive bladder cancer patients treated with radical cystectomy. The correlations of preoperative factors with ureteral involvement were analyzed by univariate analysis with Pearson's χ(2-) test and multivariate logistic regression analysis with a stepwise selection procedure. RESULTS: Positive ureteral involvement was observed in 38 (19.3%) patients. Tumor location (involvement of the vesical trigone), clinical T stage (≥ cT3) and the number of tumors (≥ 3), but not sex, tumor grade and histological features determined by transurethral resection of bladder tumor, tumor size, shape of tumor, concomitant presence of carcinoma in situ, preoperative intravesical therapy, number of transurethral resection of bladder tumor procedures or the presence of hydronephrosis were significantly associated with ureteral involvement in the univariate analysis. Multivariate logistic regression analysis confirmed that the aforementioned three significant factors identified in the univariate analysis were significant independent predictors of ureteral involvement. The probability of ureteral involvement estimated by a combination of these three parameters was well correlated with the real incidence (R = 0.904, P = 0.0021). CONCLUSIONS: Tumor location (involvement of vesical trigone), clinical T stage (≥ cT3) and the number of tumors (≥ 3) are significant independent preoperative predictors of ureteral involvement of malignancy in bladder cancer patients undergoing radical cystectomy. Our predictive model might be useful for preoperative prediction of ureteral tumor involvement.

Application of Fluolid-Orange-labeled probes for DNA microarray and immunological assays.

The usefulness of Fluolid-Orange, a novel fluorescent dye, for DNA microarray and immunological assays has been examined. Fluolid-Orange-labeled probes (DNA and IgG) were stable as examined by laser-photo-bleaching and under heat and dry conditions. Statistical analyses were performed to evaluate the reproducibility of the microarray assay, while stage-specific immunostaining of marker proteins, Kank1 and calretinin, was performed for renal cancers, both giving satisfactory results. The stability of the dye should provide advantages for storing fluorescently labeled probes and re-examining the specimens later in genetic and pathological diagnostics.

Characterization and gene expression analysis of novel matched primary and metastatic renal cell carcinoma cell lines.

Despite recent advances in molecular biology that have clarified the mechanisms involved in the metastasis of several types of cancer, the molecular mechanism underlying the metastasis of renal cell carcinoma (RCC) remains unclear. Two RCC cell lines were successfully established from the surgical specimens of a matched primary tumor and adrenal metastasis from the same RCC patient, and were designated as TMK-1P and TMK-1M, respectively. Extensive characterization was accomplished using various methods, including the Matrigel invasion assay, DNA microarray analysis and real-time reverse transcriptase (RT)-polymerase chain reaction (PCR). While TMK-1P grew faster than TMK-1M, the invasive ability of TMK-1M was higher than that of TMK-1P. DNA microarray analysis showed a large differential expression of genes related to cell adhesion and the extracellular matrix molecules of which hexabrachion (tenascin-C), epidermal growth factor receptor, cadherin-6, and beta1-catenin were down-regulated, and the 67 kDa laminin receptor 1 and transforming growth factor-beta-induced 68 kDa protein (betaig-h3) were up-regulated in TMK-1M. Real-time RT-PCR analysis confirmed this differential gene expression between the two cell lines. The RCC cell lines may be useful in studying tumor invasion and screening markers for metastasis.

Quantitative analysis of survivin mRNA expression in bladder transitional cell carcinomas.

We assessed the expression levels of survivin mRNA in bladder transitional cell carcinoma (TCC) to provide additional information regarding its malignant potential. The real-time PCR method was used to detect the survivin mRNA level for 21 bladder tumor specimens, and for urinary exfoliated cells from 12 newly diagnosed bladder tumor patients. All bladder tumor specimens and 7 of 12 voided urine specimens expressed survivin mRNA. In tumor specimens, high grade, high stage tumors had the tendency to express more survivin mRNA. Of 12 superficial bladder tumor patients who had transurethral resection of bladder tumor (TURB), 3 showed high survivin mRNA expression and intravesical recurrence after the surgery. However, for the patients who had total cystectomy due to invasive tumor, no relations were observed between the survivin mRNA expression level and development of local recurrence and/or distant metastasis. Our results suggested that the quantitative analysis of the survivin mRNA may indicate local malignant potential, which contribute to the possibility of an intravesical recurrence.

Transrectal high-intensity focused ultrasound in the treatment of localized prostate cancer: a multicenter study.

We report a multicenter trial with transrectal high-intensity focused ultrasound (HIFU) in the treatment of localized prostate cancer. A total of 72 consecutive patients with stage T1c-2NOM0 prostate cancer were treated using the Sonablate 500TM HIFU device (Focus Surgery, Indianapolis, USA). Biochemical recurrence was defined according to the criteria recommended by the American Society for Therapeutic Radiology and Oncology Consensus Panel. The median age and prostate specific antigen (PSA) level were 72 years and 8.10 ng/ml, respectively. The median follow-up period for all patients was 14.0 months. Biochemical disease-free survival rates in all patients at 1 and 2 years were 78% and 76%, respectively. Biochemical disease-free survival rates in patients with stage T1c, T2a and T2b groups at 2 years were 89, 67% and 40% (p = 0.0817). Biochemical disease-free survival rates in patients with Gleason scores of 2-4, 5-7 and 8-10 at 2 years were 88, 72% and 80% (p = 0.6539). Biochemical disease-free survival rates in patients with serum PSA of less than 10 ng/ml and 10-20 ng/ml were 75% and 78% (p = 0.6152). No viable tumor cells were noted in 68% of patients by postoperative prostate needle biopsy. Prostatic volume was decreased from 24.2 ml to 14.0 ml at 6 months after HIFU (p < 0.01). No statistically significant differences were noted in International Prostate Symptom Score, maximum urinary flow rate and quality of life analysis with Functional Assessment of Cancer Therapy. HIFU therapy appears to be minimally invasive, efficacious and safe for patients with localized prostate cancer with pretreatment PSA levels less than 20 ng/ml.

Alternative splicing of the human Kank gene produces two types of Kank protein.

The human Kank gene encodes an ankyrin repeat domain-containing protein which regulates actin polymerization. There are at least two types of Kank protein depending on cell type, likely due to differences in transcription. Here, to examine the transcriptional initiation and genomic organization of the human Kank gene, we performed 5'-RACE (rapid amplification of cDNA ends) using total RNA from normal kidney and a human kidney cancer cell line, VMRC-RCW cells. The results suggest that the human Kank gene has several alternative first exons. While mRNA from VMRC-RCW cells encoded Kank protein (referred to as Kank-S) as reported previously, mRNA from the normal kidney tissue encoded a novel type of Kank protein (referred to as Kank-L), which contained an additional N-terminal sequence 158 amino acids long. Promoter activity and the expression of the Kank variants in normal and cancer tissues were examined.

Pathological characterization of Kank in renal cell carcinoma.

The Kank gene was found as a candidate tumor suppressor gene at 9p24 by loss-of-heterozygosity search in renal cell carcinoma (RCC) and seems to have a role in controlling the formation of the cytoskeleton through the polymerization of actin. Here, we characterized the Kank protein in renal tubular cells as well as other glandular cells in the colon, stomach, prostate, testis, pancreas, thyroid, uterus, submandibular gland, adrenal, duodenum, and esophagus, and specific cells such as hepatic, alveolar myocardial, and glial cells by using a monoclonal antibody against Kank. Loss of expression of Kank in one RCC sample was detected by immunohistochemical and Western blot analyses while expression of CDKN2A (p16/Ink4A) was retained in the sample. The expression of Kank in the cytoplasm and at the sites of membrane ruffling in HEK293 and VMRC-RCW cells and in a primary culture of renal tubular cells was also detected by fluorescence-based immunostaining.

Short-term ureteral catheter stenting after uncomplicated transurethral uretero-lithotomy.

Short-term (1 or 2 post-operative days) ureteral catheter stenting after transurethral uretero-lithotomy (TUL) to avoid flank pain due to transient ureteral edema is described. Patients who underwent TUL for middle or distal ureteral stones with a rigid ureteroscope without complications during the procedures were the candidates for short-term ureteral stenting. An end-hole ureteral catheter, used to insert a guide wire during TUL, were used for stenting. The tip of the catheter was located near the renal pelvis and the other end was introduced outside through the urethra with a 14 F urethral catheter. The stent and catheter were removed on post-operative day 1 or 2. For the 18 patients treated using this method, the time of analgesic use after stent removal was 0.6+/-0.8, indicating a sufficient duration of stenting. Short-term ureteral catheter stenting is a cheap and easy way for post-operative management for uncomplicated TUL.

Metastatic malignant ameloblastoma of the kidneys.

Ameloblastoma is an uncommon disease in the urological field. The resulting tumors or cysts are of odontogenic epithelial origin, are usually benign in nature and rarely metastasize to distant organs. We describe a case of metastatic ameloblastic carcinoma in both kidneys of a 38-year-old Japanese man, who had a history of malignant ameloblastoma and was referred to us for evaluation because of gross hematuria and left flank pain. Computed tomography showed irregular cystic masses in both kidneys. After we confirmed that the primary lesion and the lung metastatic lesion had not recurred, we treated the patient surgically. Approximately 4 months postoperatively the patient suffered a local recurrence of tumors that was very invasive and aggressive. The patient died 2 months later and the autopsy showed local metastasis only, without any metastatic lesion in the lungs or other organs. The present case showed that malignant ameloblastoma is highly aggressive, and in the case of metastases the prognosis is usually extremely poor.

Cadmium nephrotoxicity and evacuation from the body in a rat modeled subchronic intoxication.

BACKGROUND: Cadmium (Cd) is an important industrial pollutant, although its mechanism of toxicity has not been completely clarified. We studied Cd-induced subchronic nephrotoxicity and the cadmium evacuation system in rats and cultured human renal tubular cells. METHODS: Male Sprague-Dawley rats were subcutaneously injected with 0.6 mg Cd/kg per day for periods of 3, 5 and 8 weeks. The concentration of Cd in urine, serum and kidneys was measured by atomic absorption spectrophotometry. Nephrotoxicity was evaluated based on the urinary concentration of beta2 microglobulin (B2MG) and histopathological findings. Apoptotic cells were detected by nick-end labeling and DNA laddering, and were based on the level of caspase-3 activity. Cadmium-induced toxicity was also studied in cultured human renal tubular cells. RESULTS: Nephrotoxicity was detected after 4 weeks of exposure to Cd, because Cd and B2MG appeared in urine. The tissue concentration of Cd increased linearly throughout the 8 weeks of exposure to Cd. The concentration of renal Cd did not change in the 3-week exposure group, but it decreased after withdrawal of Cd in the 5-week exposure group, suggesting an active Cd excretion mechanism started after the 4th week. The threshold Cd concentration for nephrotoxicity was 150 micrograms/gram wet tissue, at which concentration histological tubular damage started. Although the kidneys presented mainly necrosis, apoptosis was observed at weeks 4 and 5, before renal tubular necrosis occurred. In vitro DNA laddering was observed and peak caspase-3 activity was detected when the cells were exposed to the threshold concentration of Cd. CONCLUSION: Cadmium was effectively evacuated from the body by exfoliation of damaged renal tubular cells presenting focal tubular necrosis after the renal Cd concentration reached the threshold. Apoptosis may be involved in the regulation of Cd-induced nephrotoxicity.