Investigation of Strand-Selective Interaction of SNA-Modified siRNA with AGO2-MID.

Small interfering RNA (siRNA) has been recognized as a powerful gene-silencing tool. For therapeutic application, chemical modification is often required to improve the properties of siRNA, including its nuclease resistance, activity, off-target effects, and tissue distribution. Careful siRNA guide strand selection in the RNA-induced silencing complex (RISC) is important to increase the RNA interference (RNAi) activity as well as to reduce off-target effects. The passenger strand-mediated off-target activity was previously reduced and on-target activity was enhanced by substitution with acyclic artificial nucleic acid, namely serinol nucleic acid (SNA). In the present study, the reduction of off-target activity caused by the passenger strand was investigated by modifying siRNAs with SNA. The interactions of SNA-substituted mononucleotides, dinucleotides, and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-labeled double-stranded RNA (dsRNA) with the MID domain of the Argonaute 2 (AGO2) protein, which plays a pivotal role in strand selection by accommodation of the 5'-terminus of siRNA, were comprehensively analyzed. The obtained nuclear magnetic resonance (NMR) data revealed that AGO2-MID selectively bound to the guide strand of siRNA due to the inhibitory effect of the SNA backbone located at the 5' end of the passenger strand.

Life-threatening rupture of an external iliac artery pseudoaneurysm caused by necrotizing fasciitis following laparoscopic radical cystectomy: a case report.

BACKGROUND: Pseudoaneurysms are caused by trauma, tumors, infections, vasculitis, atherosclerosis and iatrogenic complications. In this paper, we report about a patient with rupture of an external iliac artery pseudoaneurysm, which lead to hemorrhagic shock, after undergoing laparoscopic radical cystectomy and extended pelvic lymphadenectomy. CASE PRESENTATION: The patient was a 68-year-old Japanese male diagnosed with invasive bladder cancer. Laparoscopic radical cystectomy and extended pelvic lymphadenectomy were performed. On postoperative day 12, he developed a high fever and an acute inflammatory response with redness and swelling in the right inguinal region. He was diagnosed with necrotizing fasciitis and underwent debridement. On postoperative day 42, a sudden hemorrhage developed from the open wound in the right inguinal region. He was diagnosed with external iliac artery pseudoaneurysm rupture by computed tomography. CONCLUSION: These complications occur extremely rarely after cystectomy with pelvic lymphadenectomy. There are no reports to date on these complications following laparoscopic cystectomy with pelvic lymphadenectomy.

Gender differences in the ST segment: effect of androgen-deprivation therapy and possible role of testosterone.

BACKGROUND: ST-segment elevation in a structurally normal heart is observed in Brugada- and early repolarization syndrome. The incidence of both syndromes is much higher in males than females. Clinical and basic studies suggest that testosterone plays an important role in ventricular repolarization. METHODS AND RESULTS: Standard surface 12-lead electrocardiograms recorded in 640 healthy subjects were studied (310 males, 330 females ranging in age from 5 to 89 years) (Study 1). The 3 ST levels (ST-J, -M, and -E) were measured in leads V(2) and V(5), which are representative of the right and left ventricles, respectively. The effect of androgen-deprivation therapy on the ST segment was also evaluated in 21 prostate cancer patients (Study 2). In both leads, the 3 ST levels were significantly higher in adult males than females (P<0.0001) due to a marked increase after puberty in males. As their age increased, males manifested a gradual reduction in the ST level in both leads; in females, there was a reduction in lead V(5) only. In both sexes, all 3 ST levels were significantly higher in lead V(2) than V(5) (P<0.0001). Androgen-deprivation therapy significantly decreased all 3 ST segments in both leads. CONCLUSIONS: Significant age- and gender differences in the ST segment in healthy adults were found, suggesting that testosterone modulates the early phase of ventricular repolarization.

Skeletal-related events in urological cancer patients with bone metastasis: a multicenter study in Japan.

OBJECTIVE: To investigate the incidence of skeletal-related events (SRE) in urological cancer patients with bone metastases in Japan. METHODS: Five hundred eleven patients with urological cancer and documented bone metastases treated from January 2003 to April 2008 in ten Japanese institutions were included in a retrospective analysis. Type and incidence of SRE (fracture, radiotherapy, spinal cord compression, surgery, hypercalcemia, and bone pain) were determined from patient medical records. RESULTS: The overall incidence of SRE, including 'pain', was 61%. The most common event was radiotherapy for bone metastases, with an incidence of 31%. The overall incidence of events seemed to be similar among Japanese and Western patients with prostate cancer and renal cell carcinoma when comparing data with previously published reports. Nevertheless, a much lower incidence of fracture (19.1%) was observed in Japanese renal cell carcinoma patients. CONCLUSIONS: The overall incidence of SRE in Japanese urological cancer patients with bone metastasis was similar to that in Western patients, but the incidence of fracture was lower in Japanese renal cancer patients.

A case of large solitary fibrous tumor in the retroperitoneum.

Solitary fibrous tumor (SFT) is a rare spindle cell neoplasm mainly originated in the pleural cavity. We report here an unusual case of a large SFT in the retroperitoneum. A 27-year-old female complaining of a palpable mass in the right flank with dull pain was admitted to our hospital with the diagnosis of right retroperitoneal tumor. Computed tomography (CT) and magnetic resonance imaging (MRI) showed a large retroperitoneal tumor arising from latissimus dorsi muscle. Surgical findings revealed a partly encapsulated tumor and complete resection of tumor concomitantly with right kidney, 11th and 12th ribs, and diaphragm was performed. Pathological examination demonstrated the tumor to be composed of increased mitotic activity and cellularity of spindle cells with a collagenous matrix. Immunohistochemical staining was positive for CD34, vimentin, and basic fibroblast growth factor (bFGF) and negative for CD31, cytokeratin, desmin, S-100p, smooth muscle actin, Bcl-2, and insulin-like growth factor (IGF) with Ki-67 labeling index of 0.1%. Based on pathological features, diagnosis of SFT in the retroperitoneum was confirmed. To our knowledge, this is the first report of an SFT arising from latissimus dorsi muscle and it is important to include SFT in the differential diagnosis of retroperitoneal tumors that caused considerable diagnostic problems due to its unusual site of origin.

Transitional cell carcinoma of the navicular fossa detected human papillomavirus 16.

We present a rare case of transitional cell carcinoma of the navicular fossa in an elderly man. We detected the expression of human papillomavirus type 16 specific DNA sequence in the tumor using polymerase chain reaction. Human papillomavirus dissemination into the urethra by urethral instrumentation might cause urethral tumors, such as squamous cell carcinoma or condyloma acuminatum, and also transitional cell carcinoma as seen in the present case.

Source of plasma adrenomedullin in a patient with pheochromocytoma receiving hemodialysis.

A 45-year-old man on long-term hemodialysis (HD) was incidentally discovered to have a pheochromocytoma and underwent successful resection. This patient was normotensive, and had no symptoms suggesting pheochromocytoma. The plasma concentrations of total adrenomedullin (AM-T) and mature AM (AM-m) were higher than those in normal controls. To elucidate the source of AM, we measured plasma AM levels by immunoradiometric assay before and 3 weeks after surgery in addition to plasma adrenaline, noradrenaline and dopamine. AM expression was also assessed by immunoblot and immunohistochemical analyses on normal adrenal and tumor tissues. After surgery, elevated plasma adrenaline levels returned to the normal range; however, the levels of AM-T and AM-m remained almost the same as the preoperative values. Furthermore, although AM was expressed in both normal adrenal and tumor tissues, the AM expression level was less in tumor. In this case, it was suggested that elevation in plasma AM level might be a factor associated with normotensive blood pressure; however, adrenal pheochromocytoma was not a major source of circulating AM. To our knowledge, this is the first case of pheochromocytoma in patient with HD associated with AM in the literature.

Sp1 and Sp3 transcription factors upregulate the proximal promoter of the human prostate-specific antigen gene in prostate cancer cells.

The serum level of prostate-specific antigen (PSA) is useful as a clinical marker for diagnosis and assessment of the progression of prostate cancer, and in evaluating the effectiveness of treatment. We characterized four Sp1/Sp3 binding sites in the proximal promoter of the PSA gene. In a luciferase assay, these sites contributed to the basal promoter activity in prostate cancer cells. In an electrophoretic mobility shift assay and chromatin immunoprecipitation assay, we confirmed that Sp1 and Sp3 bind to these sites. Overexpression of wild-type Sp1 and Sp3 further upregulated the promoter activity, whereas overexpression of the Sp1 dominant-negative form or addition of mithramycin A significantly reduced the promoter activity and the endogenous mRNA level of PSA. Among the four binding sites, a GC box located at nucleotides -53 to -48 was especially critical for basal promoter activity. These results indicate that Sp1 and Sp3 are involved in the basal expression of PSA in prostate cancer cells.

[Usefulness of lower ihfundibular length-to-diameter ratio as a predictor of lower pole stone clearance after extracorporeal shock wave lithotripsy; clinical research with two lithotriptors].

PURPOSE: Lower pole renal stones are well known to exhibit a poor stone clearance rate following extracorporeal shock wave lithotripsy (ESWL). In the present study, we analyzed several anatomical factors as predictors of lower pole stone clearance that may be used to indicate the usefulness and the universality of ESWL in such patients with two different lithotriptors. PATIENTS AND METHODS: 93 patients with a unilateral single lower pole stone of 2 cm. or less underwent ESWL using Piezolith 2500 or Medstone STS, were included in the study. An IVP was used to determine the lower infundibulopelvic angle, the caliceal pelvic height, the lower infundibular length, the lower infundibular diameter, the lower infundibular length-to-diameter ratio and the number of lower pole minor calyces. Stone-free status was assessed by a plain film with or without renal ultrasound. RESULTS: The stone clearance rate at the Piezolith 2500 group was 53.1% (34 of 64 patients), while was 51.7% (15 of 29 patients) at the Medstone STS group. In all cases, the overall stone clearance rate was 52.7% (49 of 93 patients). Age, laterality of the stone burden within the kidney and stone size were not different between the stone-free and residual stone groups. Multivariate logistic analysis revealed that length-to-diameter ratio was the most independent predictors of successful stone clearance at each group. The patients exhibited length-to-diameter ratio less than 7 achieved high stone clearance rates, greater than 72%. In contrast, the stone clearance rate was less than one third when length-to-diameter ratio was 7 or greater. Besides length-to-diameter ratio was strong prognostic factor in patients with stones 1 cm. or less and 1 to 2 cm at each group. CONCLUSION: From this study, it is apparent that successful ESWL is highly sensitive to the anatomy of the lower pole of the kidney. Especially, the lower infundibular length-to-diameter ratio is potentially useful and a universal predictor regardless of the kind of lithotriptors at least in patients with a lower pole radiopaque stone 2 cm. or less treated with ESWL.

Clinical characteristics of alpha-blocker responders in men with benign prostatic hyperplasia.

BACKGROUND: Currently, alpha-blocker is becoming first-line drug therapy for benign prostatic hyperplasia (BPH). Although highly effective results are obtained with this therapy, a difference between the objective and subjective response rates is reported. To prevent unnecessary medical treatment and to predict the alpha-blocker response, we characterized the clinical features of alpha-blocker responders in men with BPH. METHODS: Twenty-two men were consecutively enrolled in this study and received tamsulosin 0.2 mg once daily for 4-6 weeks. The primary measures of efficacy were maximum urinary flow rate (Q(max)) determined from the flow measurements and international prostate symptom score (IPSS). Those with an increase in Q(max) of > or = 30% from baseline and a decrease in IPSS of > or = 25% from baseline were defined as Q(max) responders and IPSS responders, respectively. Clinical findings such as age, pretreatment IPSS and Q(max), serum prostate-specific antigen (PSA), total prostate volume, transition zone (TZ) volume, TZ index and T(2)-weighted magnetic resonance image (MRI) of the prostate TZ were compared between responders and non-responders for both criteria. RESULTS: In 17 of 22 (77.2%) patients IPSS improved by > or = 25%. In 9 of 22 (40.9%) patients Q(max) improved by > or = 30%. There were no differences in clinical findings between IPSS responders and non-responders. On the contrary, Q(max) responders showed smaller total prostate and TZ volumes, a smaller TZ index and a lower intensity of the TZ in MRI than Q(max) non-responders. CONCLUSIONS: Determination of the prostate volume and MRI findings of the inner prostate gland were useful in predicting Q(max) responders to the alpha-blocker in men with BPH. In contrast, there were no clinical characteristics of the IPSS responders. IPSS responders without a Q(max) response should be closely followed while continuing the alpha-blocker therapy for a long duration.