Akihiro Morii

Early quality of life outcomes in patients with prostate cancer managed by high-dose-rate brachytherapy as monotherapy.

Objectives:  To evaluate the early quality of life outcomes in prostate cancer patients managed by high‐dose‐rate brachytherapy as monotherapy.

Effects of microbubbles on ultrasound-mediated gene transfer in human prostate cancer PC3 cells : Comparison among Levovist, YM454, and MRX-815H

We compared the enhancement effects of three different echo contrast agents (ECAs); Levovist, YM454, and MRX-815H as artificial microbubbles on ultrasound mediated gene transfection (USMGT) with 1MHz ultrasound at 0.2MPa using a luciferase expression vector in PC3 cells and elucidated the mechanisms of differences of USMGT facilitation by these ECAs. At a concentration of each ECA that induced iso-survival, ECAs with lipid shell (YM454 and MRX-815H) facilitated USMGT higher than those without shell (Levovist), and the order of the ECAs facilitating free radical formation by sonication was; YM454>MRX-815H>Levovist. These results suggested that the lipid shell type ECAs facilitated gene transfer higher than that by the non-shell type ECA.

Laparoscopic Resection of Adrenal and Extra-Adrenal Pheochromocytoma

BACKGROUND AND PURPOSE Laparoscopic resection of extra-adrenal pheochromocytoma (EAPs) necessitates meticulous surgical procedures because of changes in anatomic disposition and/or proximity to major blood vessels. Complete resection can be traumatic and may cause an increase in catecholamine levels. We present our experiences with laparoscopic resection of EAP (LEAP) and compare the intraoperative hemodynamics with those during laparoscopic resection of adrenal pheochromocytoma (LAP). PATIENTS AND METHODS We retrospectively reviewed the medical records of five patients who underwent LEAP (retrocaval EAP, n=2; interaortocaval EAP, n=1; periadrenal EAP, n=2) and five who underwent LAP between October 2001 and October 2011. We also evaluated fluctuations in blood pressure (BP) reported during both surgeries. RESULTS The tumors were successfully resected under laparoscopic guidance in both groups, and conversion to open surgery or blood transfusion was not needed. Intraoperative hypertension (BP>200 mm Hg) was observed in three LEAP and four LAP patients, whereas intraoperative hypotension (BP<80 mm Hg) was observed in five LEAP and three LAP patients. No significant differences were observed between groups, however. CONCLUSIONS Laparoscopy is the method of choice for surgeons experienced in EAP resection because it is feasible and reproducible with appropriate preoperative planning, similar to LAP.

Construction of artificial promoters sensitively responsive to sonication in vitro

PurposeTo develop artificial promoters that are activated in response to sonication and to determine these properties in vitro.MethodsThe binding sites of four transcription factors (nuclear factor-kappa B, activating protein-1, nuclear factor-Y, and CArG element binding factor A) that are activated by oxidative stress were randomly ligated and linked to a TATA-box sequence to control the luciferase gene located downstream. Transiently transfected HeLa cells from human cervical cancer with a plasmid vector containing such a gene cassette were exposed to sonication, and enhancement of luciferase expression was assessed by dual luciferase assay.ResultsOf 62 promoters constructed, two promoters, designated clone 31 and clone 62 promoters, showed a more than tenfold enhancement 6 h after sonication with 1-MHz ultrasound at 1.0 W/cm2 for 60 s. These promoters were activated in a dose-dependent manner with the intensity and duration of sonication. The activation was attenuated by addition of dimethyl sulfoxide, an antioxidant, suggesting that oxidative stress was involved. The clone 31 promoter responded to each of two serial sonications. When sonicated 24 h after the first sonication, the peak of promoter enhancement was higher than that after the first sonication.ConclusionsA promoter sensitively responsive to sonication was constructed using the above method, possibly leading to the construction of a promoter of interest that could be applied for clinical use.

Low-intensity ultrasound adjuvant therapy: enhancement of doxorubicin-induced cytotoxicity and the acoustic mechanisms involved.

PurposeIn this study, the effects of low-intensity pulsed ultrasound (LIU) as an adjuvant to doxorubicin (DOX) treatment was further investigated in comparison to hyperthermia as another widely used adjuvant. The effects were compared with respect to cell killing and apoptosis induction in U937 cells. Human primary liver cancer (PLC) cells were also used to evaluate the effects of the combinations. The use of an echo contrast agent was investigated for further enhancement of cytotoxicity. Finally, the acoustic mechanisms involved were investigated.MethodsThe effects of different treatment regimens on cell viability were determined using the Trypan blue dye-exclusion test. Apoptosis induction was detected by flow cytometry using fluorescein isothiocyanate-annexin V and propidium iodide staining. The mechanistic study involved electron paramagnetic spin trapping for detecting free radical formation as an indicator of the occurrence of inertial cavitation and spectrophotometry for sucrose hydrolysis as an indicator for noncavitational effects.ResultsThe combination treatments exerted synergistic effects on cytotoxicity depending on the acoustic conditions used. The use of LIU as an adjuvant to DOX treatment was shown to be superior to the use of hyperthermia as an adjuvant. Moreover, the combination seems to be promising for other cancer types provided that the acoustic conditions are properly selected with respect to drug concentration. The key ultrasound mechanism responsible for the synergism observed was shown to be the production of free radicals by inertial cavitation. Non-cavitational forces were also shown to contribute to the effect.ConclusionThis study is motivating to engage in in vivo research with various cancer types as a step toward clinical applicability and is emphasizing on the importance of developing therapeutic protocols for setting LIU parameters with respect to other therapeutic conditions.

Regulation of gene expression in prostate cancer cells with an artificially constructed promoter responsive to radiation.

A promoter library was developed that is composed of DNA fragments constructed by randomly elongating the cis-acting elements of transcription factors presumably activated in prostate cancer by radiation, and linking to the TATA-box sequence. One promoter with the strongest reactivity to X-ray in the LNCap cells of the library was chosen and improved by the introduction of random mutations. The resultant promoter was designated clone 880-8, showing the highest dose-dependent activity enhancement with X-ray irradiation (X-irradiation). A recombinant retrovirus expressing the luciferase gene under the control of clone 880-8 was infected into LNCap cells that showed 9.12±0.36-fold enhancement of luciferase activity 12 h after X-irradiation at 10 Gy. When the infected cells were inoculated onto nude mice, enhancement of luciferase expression was 4.27±1.36-fold 12 h after X-irradiation at 10 Gy. When LNCap was infected with another recombinant carrying the fcy::fur gene downstream from clone 880-8, fcy::fur expression was enhanced by X-irradiation. It was also shown to increase the dose-dependent cell killing ratio with 5-FC as compared with a counterpart without X-irradiation. These results suggest that the method used in this study is effective to construct a promoter responsive to stimulation. Such promoters can be used for stimulation-controlled gene therapies.

Laparoscopic Radical Nephrectomy Under Near Real-Time Three-Dimensional Surgical Navigation With C-Arm Cone Beam Computed Tomography

Objective. The aim of this study was to demonstrate the advantages of C-arm cone beam computed tomography for imaging guidance of laparoscopic radical nephrectomy (LRN). Methods. Four patients referred to the authors’ institution for LRN were included in this study. To visualize the renal vascular anatomy, the Iopamiron 300 contrast agent was injected intravenously. The surgeon could adjust the viewing angle of the images and rotate the reconstructed three-dimensional (3D) image manually by using a mouse-like controller. Using the near real-time 3D navigation images, the surgeon was able to recognize the renal vascular anatomy, and more easily perform the LRN. Results. All procedures were successfully performed with a satisfactory diagnostic yield or therapeutic effect without procedure-related complications. Conclusion. This novel technology has great potential for application in LRN because it enables accurate depiction of the renal vessels and increases surgeon confidence.

Enhancement of artificial promoter activity by ultrasound-induced oxidative stress.

We previously developed artificial promoters that were activated in response to X-ray irradiation. Sonication with 1.0MHz ultrasound that causes intracellular oxidative stress was found to activate some of these promoters though to lesser degrees. The most sensitive one among these promoters showed intensity- and duration-dependent activations by sonication. In addition, its activation by sonication was attenuated when N-acetyl cysteine was present, suggesting the involvement of intracellular oxidative stress in the activation mechanism. Improved promoters for sensitivity to X-ray irradiation were also found more sensitive to sonication. The most improved one showed 6.0 fold enhancement after sonication with 1.0MHz ultrasound at 1.0W/cm2 for 60s. This enhancement was also attenuated with the presence of N-acetyl cysteine. When stably transfected HeLa cells with the most sensitive promoter were transplanted on to mice and sonicated, luciferase activity by the promoter increased to 1.35 fold in average though it was not statistically significant compared to control. Although gene regulation in vivo by sonication was not clear, this is the first report on artificially constructed promoters responsive to ultrasound.

Laparoscopic resection of retroperitoneal neural tumors.

Purpose: Retroperitoneal neural tumor (RNT) is rarely excised laparoscopically, and the laparoscopic management of RNT remains controversial. We herein report 4 cases of laparoscopic excision of RNT that resulted in diverse clinical outcomes. Patients and Methods: Between August 2005 and January 2011, we performed laparoscopic excision of RNT in 4 patients. The mean tumor size was 4.5 cm. The mean operative time was 297 minutes and the mean amount of blood loss was 55 ml. The surgeries were uneventful, with no operative complications or evidence of intra-abdominal bleeding. However, 2 patients required reoperation for delayed hemorrhage and urinoma formation, respectively. Results: The postoperative pathological diagnoses were schwannoma in 3 patients and ganglioneuroblastoma in 1 patient. All patients were well with no signs of peripheral neuropathy or radiculopathy, and CT and/or 18F-FDG PET/CT performed during follow-up indicated no evidence of disease. Conclusions: Obtaining extensive preoperative knowledge of the source neural and vascular anatomy of the tumor is important for the surgical planning of laparoscopic resection of RNT. When a great deal of care is taken to divide the tumor and the source nerves and vital vessels, safe execution of RNT can be achieved for minimal postoperative mortality and morbidity.

Regulation of gene expression in retrovirus vectors by X-ray and proton beam radiation with artificially constructed promoters.

We previously obtained an X‐ray responsive promoter from 11 promoters that we constructed. In the present study, we aimed to determine the efficiency of our promoter construction method. In addition, the reactivity of the promoter to X‐rays in vivo is also investigated.