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Biomedical subjects

Ichiro Yoshimura

Publications and source records attributed to Ichiro Yoshimura.

15 recordsLinked to original sources

Suppression of ovarian cancer by muscle-mediated expression of soluble VEGFR-1/Flt-1 using adeno-associated virus serotype 1-derived vector.

Vascular endothelial growth factor (VEGF) is known to play a major role in angiogenesis in a variety of tumors. A soluble form of Flt-1 (sFlt-1), a VEGF receptor, is potentially useful as an antagonist of VEGF, and accumulating evidences suggest the applicability of sFlt-1 in tumor suppression by means of anti-angiogenesis. We previously demonstrated the efficacy of sflt-1 gene expression in situ to suppress tumor growth and ascites in ovarian cancer. Here, we demonstrate the therapeutic applicability of muscle-mediated expression of sFlt-1 in tumor-bearing mice. Initially, tumor suppressive action was confirmed by inoculating sFlt-1-expressing ovarian cancer (SHIN-3) cells into mice, both subcutaneously and intraperitoneally. To validate the therapeutic efficacy in a more clinically relevant model, adeno-associated virus vectors encoding sflt-1 were introduced into mouse skeletal muscles and were subsequently inoculated with tumor cells. As a result, high serum sFlt-1 levels were constantly observed, and the growth of both subcutaneously- and intraperitoneally-inoculated tumors was significantly suppressed. No delay in wound healing or adverse events of neuromuscular damage were noted, body weight did not change, and laboratory data, such as those representing liver and renal functions, were not affected. These results indicate that sFlt-1 suppresses growth and peritoneal dissemination of ovarian cancer by the inhibition of angiogenesis, and thus suggest the usefulness of gene therapy for ovarian cancer.

Animals↗

Targeting gene therapy for prostate cancer cells by liposomes complexed with anti-prostate-specific membrane antigen monoclonal antibody.

Prostate-specific membrane antigen (PSMA) is a membrane-bound antigen expressed on the surface of prostate cancer cells, and this paper describes the use of an antibody against PSMA for targeting gene therapy. We coupled anti-PSMA monoclonal antibody with poly-L-lysine and then incubated it with plasmids. These complexes were then transfected with cationic liposomes into cells. The transfection efficiency of anti-PSMA- liposome complex was higher than that of normal IgG-liposome complex in PSMA-positive LNCaP cells. Furthermore, anti-PSMA-liposome complex containing a suicide gene, thymidine kinase, demonstrated a selective growth-inhibitory effect on LNCaP cells in vitro, but did not exert a significant effect on PSMA-negative cells. In an in vivo xenograft model of LNCaP cells in nu/nu mice, we administered the complexes via the tail vein. Judging on the basis of both 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining and luciferase assay findings, a significant enrichment of plasmid DNA was observed in LNCaP xenografts with anti-PSMA-liposome complex in comparison with normal IgG-liposome complex. However, the distribution of plasmid DNA did not change substantially in any other organs including the liver, kidney, lung, and spleen. Moreover, in suicide gene therapy, anti-PSMA-liposome complex exerted a significant inhibitory effect on the growth of LNCaP xenograft, in contrast to normal IgG-liposome complex.

Animals↗

Partial physeal growth arrest treated by bridge resection and artificial dura substitute interposition.

We treated four cases of partial physeal growth arrest by resecting the bone bridge and interposing an expanded polytetrafluoroethylene membrane, which has minimal tissue response. We evaluated three cases with sufficiently long follow-up. No apparent recurrence of the bone bridge and no limb length discrepancy occurred in any of the cases. No remodeling after the operation occurred. In one case, which underwent this operation and corrective osteotomy of the proximal tibia, slight recurrence of the varus deformity occurred which was detected only by radiographs, and did not compromise the alignment of the leg. There were no complications.

Bone Diseases, Developmental↗

Retroperitoneoscopic partial nephrectomy using radiofrequency ablation.

Two patients with renal tumors underwent retroperitoneoscopic partial nephrectomy. The renal tumors were initially treated with radiofrequency ablation. This method allowed tumor excision to be achieved without clamping the renal pedicle. Residual renal function was well maintained as determined by enhanced computed tomography scanning and measurement of the serum creatinine level. There were no complications such as vascular damage or collecting system injury. The pathological diagnosis was clear cell carcinoma (pT1) in Patient 1 and was not determined in Patient 2 because of entire ablation. No recurrence has been observed after 3 years and 2 years of follow up, respectively. Radiofrequency ablation was useful for control of local bleeding during retroperitonaoscopic partial nephrectomy.

Adenocarcinoma, Clear Cell↗

Mucoid pseudocyst of the obturator nerve.

A 71-year-old man visited the outpatients' clinic of National Saitama Hospital, Japan, complaining of pain in the right leg. Magnetic resonance imaging (MRI) revealed a cystic mass adjacent to the right side of the urinary bladder. He underwent the removal of the cystic mass and the pathological diagnosis was a mucoid pseudocyst of the obturator nerve. After tumor resection, the pain was diminished and he has been free of disease for 3 years.

Aged↗

Selective gene therapy for prostate cancer cells using liposomes conjugated with IgM type monoclonal antibody against prostate-specific membrane antigen.

Prostate cancer cells express prostate-specific membrane antigen (PSMA). We developed an IgM type monoclonal antibody against PSMA. The antibody was coupled to poly-L-lysine and thereafter this conjugate was mixed with cationic liposomes containing plasmid DNA. The antibody-liposome complex was tested whether it could deliver the gene of interest selectively to the PSMA positive cells. As assessed by beta-galactosidase reporter gene, the transfection efficiency was 13.2% with anti-PSMA-liposome complex as compared to 4% with control IgM liposome complex. In contrast, no such differences were observed in PSMA negative PC-3, DU145 and T24 cells. Furthermore, in the suicide gene therapy in vitro with thymidine kinase gene plus ganciclovir system, anti-PSMA liposome complex demonstrated a selective growth inhibitory effect on PSMA positive LNCaP cells but not on PSMA negative cell lines.

Adenocarcinoma↗

Suppression of lung metastasis of renal cell carcinoma by the intramuscular gene transfer of a soluble form of vascular endothelial growth factor receptor I.

PURPOSE: We clarified whether adenovirus mediated intramuscular gene transfer of soluble vascular endothelial growth factor receptor 1 (sFlt-1) can inhibit lung metastasis of renal cell carcinoma. MATERIALS AND METHODS: We constructed the adenovirus vector AdsFlt-1, which expresses soluble Flt-1 protein. Functional validation of the vector was determined by HUVEC (in vitro human umbilical vein endothelial cell) proliferation inhibition assay. The efficacy of AdsFlt-1 was tested in a Renca murine renal cell carcinoma lung metastasis model. BALB/c mice were injected with Renca cells, followed by the intramuscular administration of AdsFlt-1 24 hours later. Lung specimens were harvested 17 days later and the number of lung metastases was counted. Immunohistochemical analysis of the specimen for angiogenesis and apoptosis was done. RESULTS: Treatment with adenovirus expressed sFlt-1 inhibited HUVEC proliferation. The intramuscular administration of AdsFlt-1 significantly inhibited lung metastasis of Renca cells. Immunohistochemical analysis of the lung specimen showed fewer neovessel formations and more apoptotic cells in AdsFlt-1 treated tumors in mice. CONCLUSIONS: The intramuscular administration of AdsFlt-1 effectively inhibited lung metastasis in a murine model of renal cell carcinoma. Because of the simplicity of this therapy, it may well be useful as an effective adjuvant therapy for renal cell carcinoma.

Adenoviridae↗

Treatment efficiency of a suicide gene therapy using prostate-specific membrane antigen promoter/enhancer in a castrated mouse model of prostate cancer.

Suicide gene therapy has potential for the treatment of prostate cancer under conditions of androgen deprivation. We show here that the combination of promoter/enhancer of prostate-specific membrane antigen (PEPM) and the Cre-loxP system is a good method to express a suicide gene, namely herpes virus thymidine kinase (TK), in prostate cancer cells. We have examined this system in a castration model in vivo, in comparison with a prostate-specific antigen promoter/enhancer system (PP). In the castrated mice, the tumor luciferase activity with the combination of the PEPM plus the Cre-loxP system was about 50 times greater than that with the control GL3 plasmid. A similar increase was observed in non-castrated mice. In contrast, the luciferase activity of the plasmid PP was decreased significantly in tumors from castrated mice as compared with tumors from non-castrated control mice. Regarding the therapeutic effect, the combination plasmid PEPM-Cre plus CMV-loxP-TK exhibited a strong inhibitory effect on tumor growth in the castrated mice, as in the non-castrated mice. In contrast, PP-TK plasmid did not show any significant growth inhibition in the castrated mice. These findings indicate that the combination of PEPM and Cre-loxP system may have a good treatment effect under androgen ablation conditions in vivo, and our system may therefore be applicable to patients who have previously received androgen deprivation therapy.

Adenocarcinoma↗

Effects of CD/5-FC suicide gene therapy system on human malignant glioma cells in vitro.

The purpose of this paper is to investigate the antitumor effects of cytosine deaminase/5-fluorocytosine (CD/5-FC) suicide gene therapy system on human malignant glioma cells in vitro. The pCMVCD plasmid was constructed through the CD gene insertion in the multicloning site of eukaryotic expression vector pcDNA3.0, and confirmed by restriction endonuclease digestion/gene sequencing. The construct was subsequently transfected into the U251 human malignant glioma cells by using LipofectAMINE2000-mediated method. Resistant clones (named U251/CD cells) were isolated by screening with G418 presence. U251/CD cells were incubated with 5-FC in different concentrations to determine viability ratios (or cytotoxicity assay), measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The concentrations of 5-fluorouracil (5-FU) in the media were measured by high-performance liquid chromatography (HPLC) detector. Our results suggested that the untreated U251 cells were insensitive to 5-FC, with the IC(50) about 6500 micromol/L. After transfection, the IC(50) was dramatically reduced to about 10 micromol/L. Therefore, gene transfection made G418-resistant clones (U251/CD cells) be highly sensitive to 5-FC. HPLC analysis showed that 5-FU was detected in U251/CD cell medium. Study on U251 cells genetically modified by CD gene in vitro will play an essential role in glioma gene therapy in vivo. In conclusion, our results indicated that the CD/5-FC system was feasible to treat glioma.

Cell Death↗

Lateral thrust of anterior cruciate ligament-insufficient knees and posterior cruciate ligament-insufficient knees.

Leaving anterior cruciate ligament (ACL) insufficiency and posterior cruciate ligament (PCL) insufficiency untreated frequently leads to osteoarthritis (OA). The purpose of this study was to evaluate dynamically the lateral thrust of ACL-insufficient knees and PCL-insufficient knees, and from the findings investigate the relationship between cruciate ligament insufficiency and OA occurrence. An acceleration sensor was attached to the affected and control anterior tibial tubercles, acting in medial-lateral and perpendicular directions. The lateral thrust immediately after heel strike was measured continuously by a telemeter under stabilised walking conditions. When compared to the contralateral healthy knee, the peak value of lateral acceleration immediately after heel strike was significantly larger in the ACL-insufficient knee; and lateral thrust was increased, but not significantly, in the PCL-insufficient knee. Given that lateral thrust of the knee during walking increases due to ACL or PCL injury, it may be a principal contributor to OA progression.

Adolescent↗

Can initial prostate specific antigen determinations eliminate the need for bone scans in patients with newly diagnosed prostate carcinoma? A multicenter retrospective study in Japan.

BACKGROUND: The objective of the current study was to assess rigorously whether serum prostate specific antigen (PSA) determination can eliminate the need for bone scans in Japanese patients with newly diagnosed prostate carcinoma with serum PSA levels < or = 10 ng/mL. METHODS: A retrospective assessment of 1294 patients with newly diagnosed, untreated prostate carcinoma was conducted at the authors' institutions. All patients underwent a bone scan, serum PSA measurement, and core needle biopsy of the prostate. The receiver operating characteristic curve for identifying a positive bone scan based on serum PSA levels and a decision tree were analyzed to determine the expected 10-year cumulative cost and disease specific survival rate. Two competing strategies were used: PSA alone and PSA plus baseline bone scan. For the PSA-alone strategy, a baseline bone scan was performed only when the patient had a serum PSA level > 10 ng/mL. RESULTS: The proportion of positive bone scans in patients with serum PSA levels < or = 10.0 ng/mL was 1.33%. The area under the receiver operating characteristic curve was 0.870. Patients with a Gleason Grade > or = 3 tumors or with a Gleason score > or = 7 had a higher proportion of positive bone scans. The 10-year disease specific survival rates with the PSA-alone strategy and the PSA-plus-bone-scan strategy were the same. The PSA-alone strategy was minimally cost effective, with a savings of $16.00 (U.S.) in the cumulative net cost per patient over the PSA-plus-bone-scan strategy. CONCLUSIONS: The current results suggest that baseline bone scans can be eliminated in patients with newly diagnosed prostate carcinoma in Japan who have serum PSA levels < or = 10 ng/mL. Apparently, it is possible to omit baseline bone scans for patients with a Gleason Grade < or = 2 tumors or with a Gleason score < or =6.

Adult↗

Adenovirus mediated prostate specific enzyme prodrug gene therapy using prostate specific antigen promoter enhanced by the Cre-loxP system.

PURPOSE: Tissue or tumor specific gene delivery is crucial for achieving successful results in suicide gene therapy. Prostate specific antigen (PSA) promoter is known to be highly specific in prostate tissue but its promoter activity is much weaker than that of constitutive viral promoters. In the current study we enhanced PSA promoter activity by combining it with the Cre-loxP system. We also applied this system to adenovirus mediated suicide gene therapy with the cytosine deaminase (CD) gene. MATERIALS AND METHODS: The Cre-loxP DNA recombination system was used to enhance PSA promoter. A plasmid with the PSA promoter-enhancer combination was constructed to drive Cre recombinase. Another plasmid contained the cytomegalovirus promoter-loxP-flanked stop signal-luciferase gene. LNCaP human prostate cancer cells were co-transfected with these 2 plasmids and luciferase activity was measured to assess promoter activities. Adenoviral vectors with the CD suicide gene were constructed in similar fashion and tested in LNCaP cells in in vitro/in vivo prostate cancer models. RESULTS: Promoter activity of the combined PSA promoter/enhancer and Cre-loxP system was 3 times stronger than that of PSA promoter/enhancer alone. It was further enhanced 7-fold in the presence of testosterone. Application of this system to CD suicide gene therapy by adenoviral vectors inhibited subcutaneous LNCaP tumor growth in nude mice. CONCLUSIONS: Combining the Cre-loxP system with PSA promoter/enhancer amplified promoter activity and was found to inhibit the growth of PSA producing prostate cancer cells in vivo.

Adenoviridae↗

The influences of biomechanical factors on cartilage regeneration after high tibial osteotomy for knees with medial compartment osteoarthritis: clinical and arthroscopic observations.

PURPOSE: A high tibial osteotomy (HTO) is often performed on knees with medial compartment osteoarthritis. However, in varus knees with medial compartment osteoarthritis, degeneration of the articular cartilage tends to progress due to medial deviation of the mechanical axis. This can result in eburnated bone. A widening of the medial joint space has been observed after HTO with a proper correction as a result of decreased stress on the load-bearing cartilage in the medial compartment. Researchers have reported obtaining a repair of the articular cartilage with both eburnation and clinical improvement using HTO in patients showing a medial deviation of the mechanical axis. However, which factors influence cartilage regeneration remain unclear. The purpose of this study was to quantitatively evaluate the influence of cartilage regeneration after a high tibial osteotomy for knees with medial compartment osteoarthritis. TYPE OF STUDY: Observational cohort study. METHODS: The study involved 58 knees in 47 patients. A modified Noyes classification was used for arthroscopic grading. The patients underwent a second look arthroscopic evaluation of the articular cartilage 18 months after surgery. The articular cartilage was classified as no regenerative change (grade 1), white scattering with fibrocartilage (grade 2), partial coverage with fibrocartilage (grade 3), and even coverage with fibrocartilage (grade 4). The functional results were evaluated according to the Japanese Orthopedic Association score for osteoarthritic knees (JOA score), femorotibial angle (FTA), percentage of mechanical axis, and body mass index (BMI). RESULTS: Partial or even coverage with fibrocartilage (grade 3 and 4) was achieved on 55% of the femorotibial joint surfaces. A repair with white scattering with fibrocartilage (grade 2) was achieved in 34%, and 3 knees showed no regenerative change (grade 1). At follow up, significant differences were seen between grade 4 and grade 1 in JOA score and percentage of mechanical axis. CONCLUSIONS: This study demonstrates the correlation between the visible improvement of the articular surface, the functional score, and the degree of correction obtained with a lateral closing wedge HTO.

Aged↗

Development of gene therapy using prostate-specific membrane antigen promoter/enhancer with Cre Recombinase/LoxP system for prostate cancer cells under androgen ablation condition.

To enhance the efficacy of suicide gene therapy for prostate cancer under androgen deprivation, we designed a promoter system that consists of the prostate-specific membrane antigen (PSMA) promoter / enhancer (PEPM) and Cre-loxP DNA recombination system. We constructed two kinds of plasmids. One plasmid contains a Cre recombinase (Cre) under the control of PEPM and the other expresses CMV-lox-luciferase / herpes simplex virus thymidine kinase (TK). In PSMA-positive LNCaP cells, the promoter activity of the PEPM-Cre plus CMV-lox-luciferase demonstrated 800-fold greater activity compared with that of the PSMA promoter alone. However, no enhancement of the promoter activity was observed in the PSMA-negative cells. Furthermore, in contrast to prostate specific antigen promoter / enhancer (PP), the promoter activity of PEPM did not decrease when the LNCaP cells were cultured in charcoal-stripped fetal bovine serum (CFBS). In an in vitro gene therapy model with LNCaP cells, the cell growth inhibition in the presence of ganciclovir (GCV) was more evident in the cells transfected with the PEPM-Cre plus CMV-lox-TK than in the cells with the PP-TK, and the difference in efficacy between the two plasmids was more remarkable when the cells were maintained in CFBS medium. The therapeutic effect of PEPM-Cre plus CMV-lox-TK was also observed in xenografted LNCaP cells on nude mice when the plasmids were directly injected into tumors and GCV was administered intraperitoneally. These findings indicate that the combination of the PSMA promoter / enhancer and the Cre-loxP system can enhance the PSMA promoter activity even under androgen ablation conditions and can exert its anti-tumor effect both in vitro and in vivo.

Animals↗

The effect of wedged insoles on the lateral thrust of anterior cruciate ligament-insufficient knees.

BACKGROUND: Patients with untreated anterior cruciate ligament insufficiency often develop osteoarthritis. In Japan, laterally or medially elevated insoles have been widely used for the relief of symptoms associated with unicompartmental osteoarthritis of the knee. HYPOTHESIS: The use of a valgus wedge in the shoe of a patient with anterior cruciate ligament insufficiency will decrease lateral thrust. STUDY DESIGN: Controlled laboratory study. METHODS: An accelerometer was used to measure thrust force in 60 control knees and 35 knees with anterior cruciate ligament insufficiency. The sensor was attached to the affected and the control anterior tibial tubercles to record motion in the medial-lateral and perpendicular directions. The lateral thrust immediately after heel strike was measured continuously by a telemeter under stabilized walking conditions. Measurements were made both with and without wedged insoles. RESULTS: When compared with the control knee, the peak value of lateral acceleration immediately after heel strike was significantly greater in the anterior cruciate ligament-insufficient knees. However, lateral thrust decreased significantly in the anterior cruciate ligament-insufficient knees when a wedged insole was used. CONCLUSIONS: Use of an insole resulted in a significant decrease in lateral thrust, even in anterior cruciate ligament-insufficient knees. CLINICAL RELEVANCE: Use of an insole is one possible prophylaxis for osteoarthritis for an anterior cruciate ligament-insufficient knee.

Acceleration↗