Gene therapy of ischemic-damaged kidney in the rat using hepatocyte growth factor gene.
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Animals↗
Biomedical subjects
A Okuyama
Publications and source records attributed to A Okuyama.
Strong inhibition of crystal-cell attachment by pediatric urinary macromolecules: a close relationship with high urinary citrate secretion.
OBJECTIVES: To investigate other reasons for the low incidence of pediatric urolithiasis, we evaluated the difference in the crystal-cell adhesion inhibitory activity of urinary macromolecules (UMMs) between children and adults. We also evaluated whether citrates influence the above inhibitory activity, because citrates are important in pediatric urine. METHODS: Urine samples were collected from children and healthy male adults during a 24-hour period, and urinary components with a molecular weight of 3 kDa or greater were extracted as UMMs to compare their inhibitory activity during the adhesion of calcium oxalate monohydrate crystals to cultured Madin-Darby canine kidney cells between children and adults. Subsequently, various concentrations of citrates were added to adult UMMs to evaluate the changes in the crystal-cell adhesion inhibitory activity of UMMs. RESULTS: Pediatric UMMs more strongly inhibited the adhesion of calcium oxalate monohydrate crystals to cultured Madin-Darby canine kidney cells at a concentration of 0.1 mg/mL compared with adult UMMs. In addition, pediatric UMMs contained higher proportions of fibronectin and glycosaminoglycans, both of which exhibit crystal-cell adhesion inhibitory activity. When citrates were added to adult UMMs, the crystal-cell adhesion inhibitory activity of UMMs was increased in a dose-dependent manner. However, citrates alone did not result in any differences in the inhibitory activity at any of the three different concentrations. CONCLUSIONS: We speculate that the incidence of pediatric urolithiasis is low because pediatric UMMs more potently inhibit the adhesion of calcium oxalate crystals to renal tubular cells or because the higher proportion of citrates in pediatric urine enhances the crystal-cell adhesion inhibitory activity of UMMs in a dose-dependent manner.
Adult↗
Effects of hepatocyte growth factor on E-cadherin-mediated cell-cell adhesion in DU145 prostate cancer cells.
OBJECTIVES: To examine how hepatocyte growth factor (HGF) affects cell-cell adhesion junctions on scattering in prostate cancer cells. HGF is known to induce scattering (dispersion of clustered cells into single cells) in various epithelial cells, including prostate cancer cells, but the mechanisms surrounding this action are not fully understood. Cell-cell adhesion junctions are composed of E-cadherin and its associated intracellular catenins and play important roles in the maintenance of cell integrity. METHODS: The human prostate cancer cell line DU145 was used in this study. The associations and changes of various adhesion molecules with HGF treatment were investigated by inhibition assays, Western blot analysis, and immunofluorescence staining. RESULTS: In the inhibition assay, anti-E-cadherin neutralizing monoclonal antibody caused the dissociation of DU145 cells similar to the scattering with HGF treatment. The expression of E-cadherin decreased with HGF, and the expression of alpha-catenin and beta-catenin did not change by Western blot analysis. In immunofluorescence staining, HGF caused the translocation of E-cadherin from cell-cell adhesion junctions to the cytoplasm. CONCLUSIONS: These results indicate that HGF induces scattering by decreasing the expression of E-cadherin and causes its translocation to the cytoplasm of DU145 cells.
Cadherins↗
Rapid and simple determination of mycophenolic acid in human plasma by ion-pair RP-LC with fluorescence detection.
Mycophenolic acid (MPA) is an immunosuppressive drug given as the prodrug of mycophenolate mofetil (MMF). In order to investigate the pharmacokinetics of MPA, a simple, specific, sensitive and reliable method has been established for the quantitative determination of MPA in plasma from renal transplant recipients. The method involves a single-step protein precipitation procedure and a specific determination by ion-pair reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Separation was achieved on a C18 column (150 x 4.6 mm, 5 microm) with a mobile phase composed of borate buffer (pH 10.0; 50 mM)--acetonitrile--tetrabutylammonium bromide (200 mM) (75:25:1, v/v/v). The fluorescence detector was set at 310 (excitation) and 430 nm (emission). Following protein precipitation with ice-cold acetonitrile, clear supernatants (50 microl) were injected into the HPLC system. The retention time of MPA was approximately 4.5 min. The HPLC run time was 8 min. The assay was linear in concentration range 0.2-20.0 microg/ml for MPA in human plasma. Precision of the assay in the concentration range examined was from 0.89 to 3.21% for the intra-assay run and from 3.01 to 4.35% for the inter-assay run. A limit of detection was 0.05 microg/ml at a signal-to-noise ratio of 3. This validated method was then applied to the determination of MPA concentrations in renal transplant recipients after oral administration of 0.75 g of MMF.
Chromatography, High Pressure Liquid↗
The differences between late graft loss group and long-term graft survival group in renal transplantation.
In renal transplantation, the long-term graft survival rate has not been improved. Until now, the differences between late graft loss and long-term graft survival have still not been estimated thoroughly. We have attempted to define clinical risk factors and parameters for late graft loss by comparing the differences in these two groups. Data from the Osaka University Database were assessed on 156 renal allografts during a 7-yr period. Thirty-six patients comprised the late graft loss group (patients in this group had graft function without need for dialysis for more than 3 yr post-transplantation, afterwards lost the allograft: 'loss group'). One hundred and twenty patients comprised the long-term graft survival group (patients in this group had graft function without need for dialysis until 31 December 1999: 'survival group'). Various immunological and non-immunological parameters were included in an univariate regression analysis. This analysis showed that donor age (P < 0.01), HLA mismatch number (P < 0.01) and a repeat of acute rejection (P < 0.01) were significant factors. Serum creatinine levels at 3 months (P = 0.01), proteinuria at 1 yr (P < 0.01) and antihypertensive treatment at 2 yr (P = 0.03) after transplantation were predictive of the risk of late graft loss. CsA trough concentration at 3-6 months (P < 0.05) and body mass index increase at 1 yr (P = 0.046) were elevated in the loss group. These results from a single centre suggest that immunological as well as non-immunological factors are associated with the pathogenesis of late graft loss.
Adult↗
A case of tacrolimus nephrotoxicity appearing in a second renal transplantation patient.
We experienced a case of a second renal transplantation patient. With the use of cyclosporin, he lost his first graft because of chronic rejection; with the use of tacrolimus, his second graft suffered from drug nephrotoxicity. On his second renal transplantation, his graft function deteriorated and required haemodialysis with the use of tacrolimus. Repeated biopsies did not reveal the typical characteristics of acute tacrolimus nephrotoxicity and acute rejection. His tacrolimus trough level was not high during the clinical course; however, by reducing tacrolimus dosage, his graft function eventually recovered to mild renal dysfunction. This observation was helpful for clinical diagnosis of the functional toxicity of tacrolimus. The case is interesting in considering the functional toxicity of tacrolimus and the difference between tacrolimus and cyclosporin in terms of immunosuppressive and nephrotoxic actions.
Biopsy↗
Fas gene mutations in prostatic intraepithelial neoplasia and concurrent carcinoma: analysis of laser capture microdissected specimens.
Fas (Apo-1/CD95) is a cell-surface receptor involved in cell death signaling through binding of Fas ligand. Mutations of the Fas gene might be involved in proliferative diseases of the prostate by prolongation of programmed cell death of prostatic epithelial cells. Using the laser capture microdissection method, Fas gene mutations were examined on genomic DNA extracted from lesions with high-grade prostatic intraepithelial neoplasia (HGPIN), a possible precursor of prostatic cancer (PCA), and from PCA. A total of 193 lesions, 111 with HGPIN, 55 with PCA, and 27 benign glands, were microdissected from 27 patients with PCA. Polymerase chain reaction-amplified products were directly sequenced. Loss of heterozygosity (LOH) was examined at four sites of known polymorphisms. Fas gene mutations were detected in HGPIN: 4 of 27 (14.8%) cases or 4 of 111 (3.6%) lesions. All were point mutations: three missense and one nonsense in the death domain. Benign proliferative glands adjoining HGPIN and/or PCA, and PCA never showed mutations. LOH was found in 31.3% of PCA and 25% of HGPIN lesions, but was never found in benign glands. Exclusive occurrence of Fas mutations in HGPIN might underlie the development of these lesions. Occasional findings of LOH in HGPIN and PCA suggested that genetic instability might occur during the early phase of prostatic carcinogenesis.
Alleles↗
A new matrix metalloproteinase inhibitor SI-27 induces apoptosis in several human myeloid leukemia cell lines and enhances sensitivity to TNF alpha-induced apoptosis.
MMP inhibitors are used clinically for the stabilization of tumor growth, thus prolonging survival in cancer patients. However, their role in the treatment of hematopoietic malignancies remains unclear. In the present study, we investigated the effects of a new MMP inhibitor, SI-27, in hematopoietic malignancies. SI-27 alone induces apoptosis in several human myeloid leukemia cell lines such as U937, NB4, and HL60 cells by activating caspase 8, 9, and 3. Apoptosis was measured with annexin V positive staining, a drop in mitochondrial transmembrane potential (deltapsim), presence of hypodiploid DNA, and cleavage of PARP and IkappaBalpha. Furthermore, at lowered concentrations, which did not directly induce apoptosis, SI-27 acted to sensitize U937 cells and other cells to tumor necrosis factor alpha (TNF-alpha)-mediated apoptosis. The accumulation of membrane Fas, the Fas ligand, and TNFR1 were not apparent due to exposure to SI-27, and antagonistic anti-Fas or anti-Fas ligand antibodies did not block SI-27-induced apoptosis. Thus, SI-27-induced apoptosis is not mediated by the Fas pathway. These results suggest that MMP inhibitors, alone or in combination with other cytotoxic agents, can provide a unique method for treating acute myeloid leukemia, refractory to classical anti-cancer drugs, and may thus suppress recurrence.
Apoptosis↗
Chromosomal variants among 1790 infertile men.
BACKGROUND: The largest cytogenetic survey involving infertile men was undertaken to clarify whether chromosomal abnormalities, including autosomal abnormalities, affect semen qualities. METHOD: All male patients who visited an infertility clinic from 1990 to 1998 underwent chromosomal and semen analysis. RESULTS: Chromosomal abnormalities were found in 225 of 1790 patients (12.6%). The most frequent anomaly was Klinefelter syndrome (64 cases). Autosomal anomalies accounted for 126 cases. 46,XY,1qh(+) was the most common autosomal anomaly (30 cases) and its incidence was significantly higher than those of normal controls. The seminograms of these patients varied widely, with nine patients having azoospermia and three patients achieving natural pregnancies. It is not yet clear if this karyotype affects spermatogenesis. CONCLUSION: Autosomal anomalies as well as sex chromosomal abnormalities might affect spermatogenesis. Cytogenetic study is important before intracytoplasmic sperm injection.
Chromosome Aberrations↗
Evaluation of the efficacy and safety of laparoscopic nephrectomy.
PURPOSE: To evaluate the efficacy and safety of laparoscopic nephrectomy. METHODS: From June 1994 to November 1999, 10 patients underwent laparoscopic nephrectomy at Osaka University Medical Hospital and Osaka Rosai Hospital. Laparoscopic nephrectomy was performed either via transperitoneal or retroperitoneal approach under general anesthesia. These 10 cases were reviewed in respect of primary disease of the kidney, operative time, complications and postoperative convalescence. RESULTS: Of the 10 patients, five were preoperatively diagnosed as having a non-functioning kidney with hydronephrosis, two patients were diagnosed as having an atrophic kidney, two had renal cell carcinoma and one had renal pelvic tumor. The average operative time was 374 min (range 270-675 min). The mean blood loss was 330 mL (range 60-800 mL). One patient required transfusion due to postoperative oozing. The average hospital stay after operation was 7 days. No major postoperative complications were observed. CONCLUSION: Laparoscopic nephrectomy is an option in surgically managing renal disorders, including malignancies, although it has a longer operative time compared to conventional open surgery.
Adult↗
Retrovesical desmoplastic small round cell tumor in a patient with urinary frequency.
A 21-year-old man with urological symptoms was found to have a large abdominal tumor in the retrovesical space between the bladder and the rectosigmoid colon. Radiologic findings revealed little evidence confirming the diagnosis. A transrectal biopsy failed to disclose the histopathologic origin of the tumor. An exploratory laparotomy with a complete surgical resection was impossible, so a wedge biopsy was performed. Combined histologic and immunohistochemical findings revealed the features of desmoplastic small round cell tumor (DSRCT). Despite subsequent multi-agent chemotherapy, the patient died as a result of the growing tumor and liver metastasis. There have been only two prior reports of this neoplasm in the urological literature.
Abdominal Neoplasms↗
Early stage small cell carcinoma of the urinary bladder.
A 73-year-old man with primary small cell carcinoma of the bladder underwent radical cystectomy. The pathological findings revealed the tumor confined to the submucosal layer (pT1) without metastasis. No adjuvant chemotherapy was carried out. He is alive with no evidence of the disease 24 months after the operation.
Aged↗
Laparoscopic diagnosis and treatment of nonpalpable testis.
OBJECTIVE: Laparoscopy has become one of the important diagnostic modalities of nonpalpable testis and has been developed and applied in the treatment of this disease. In the present study, we investigated the usefulness of laparoscopy in the diagnosis and treatment of nonpalpable testis. METHODS: Laparoscopy was carried out under general anesthesia on 21 patients (23 testes) from October 1991 to October 1999. If the internal spermatic vessels and vas deferens made their way into the internal inguinal ring, the inguinal canal was dissected with an incision in the inguinal region to look for the testis. Patients with intra-abdominal testis underwent laparoscopic orchiopexy or orchiectomy. If the internal spermatic vessels terminated with a blind end intraperitoneally, making it impossible to identify the testis, the case was judged to be vanishing testis and the operation was finalized without any further examination. RESULTS: In eight of 23 testes (35%), the internal spermatic vessels and vas deferens made their way into the internal inguinal ring. The inguinal region was examined in all the eight testes. Orchiopexy was carried out on two testes and orchiectomy was carried out on six testes. An intra-abdominal testis was detected in eight of 23 testes (35%). Laparoscopic orchiopexy was carried out on seven testes. One-stage orchiopexy was carried out on two of the seven testes and two-stage Fowler-Stephens orchiopexy was carried out on five of the seven testes. Orchiectomy was carried out on the remaining testis. Blocking or lack of the internal spermatic vessels and vas deferens was seen in seven of the 23 testes (30%) and this condition was diagnosed as vanishing testis. CONCLUSION: Laparoscopy for nonpalpable testis is considered to be the most effective technique for diagnosing the presence or absence of the testis and the location of the testis.
Adolescent↗
Clinical evaluation of human telomerase catalytic subunit in bladder washings from patients with bladder cancer.
PURPOSE: We examined the expression of mRNA of human telomerase reverse transcriptase (hTERT), a catalytic subunit of the telomerase complex, in bladder washings as a tumor marker for the detection of bladder cancer. MATERIALS AND METHODS: Bladder washings were obtained from 63 patients. We examined the expression of hTERT mRNA by the nested reverse transcription-polymerase chain reaction (RT-PCR) method and also measured the relative expressions of hTERT mRNA and beta(2)-microglobulin (beta(2)-MG) in these samples by RT-PCR analysis. Correlations between the relative expression of hTERT mRNA and clinical features were analyzed. We also compared the sensitivity of this assay with that of urinary cytology. RESULTS: By nested RT-PCR, we detected three false-positive cases (11%) in the control group. Therefore, the relative expression values of hTERT mRNA and beta(2)-MG correlated strongly with tumor size, but not with multiplicity or histologic grade. When the cut-off value of the expression was fixed at 0.27%, the sensitivity and specificity of this assay were 74% and 93%, respectively. This assay was more sensitive than urinary cytology for the detection of bladder cancer. CONCLUSION: These results suggest that the relative expression of hTERT mRNA in bladder washings is useful in screening for bladder cancer. Relative expression is of assistance in diagnosing bladder cancer.
Biomarkers, Tumor↗
Loss of p73 induction in a cisplatin-resistant bladder cancer cell line.
BACKGROUND: Cisplatin (CDDP) plays an important role in the treatment of transitional-cell carcinoma (TCC), but resistance develops. The mechanism is not entirely understood. METHODS: To assess acquired resistance to CDDP, we established a CDDP-resistant subclone, CL8-2, of T24, which is a bladder cancer cell line. We examined the changes in the various pathways leading to apoptosis in the parent line and CL8-2. RESULTS: The drug caused apoptosis of T24 cells but not CL8-2 cells. The CL8-2 cells were 6.4 times more resistant to CDDP than was T24. In both cell lines, the mismatch repair genes hMLH-1 and hMSH-2 were expressed at high levels. The p53 protein was not detected in either cell line but p73 protein was induced by CDDP treatment in T24 cells, which was followed by activation of caspases 3, 8, and 9. This phenomenon was not observed in CL8-2 cells. CONCLUSION: These results suggest that loss of p73 induction may lead to CDDP resistance of TCC.
Antineoplastic Agents↗
Testosterone suppresses spermatogenesis in juvenile spermatogonial depletion (jsd ) mice.
Male juvenile spermatogonial depletion (jsd/jsd) mice are sterile because of a failure of spermatogonial differentiation. We have previously reported the recovery of spermatogonial differentiation by suppressing the levels of gonadotropins and testosterone with Nal-Glu, a GnRH antagonist. To determine whether suppression of testosterone or the gonadotropins was responsible for spermatogenic recovery, we examined the effect of supplementation of LH or FSH along with Nal-Glu treatment. Systemic administration of flutamide, an androgen receptor antagonist, was also examined. LH supplementation elevated both serum and intratesticular testosterone levels and suppressed the recovery of spermatogonial differentiation in a dose-dependent manner. Supplementation with FSH did not affect either testosterone levels or spermatogonial differentiation. Furthermore, the mice treated with flutamide showed some recovery of spermatogonial differentiation. The overall findings revealed that testosterone action mediated by androgen receptors suppressed the spermatogonial differentiation in jsd/jsd mice and suggested that spermatogonial differentiation in the jsd mutant is highly sensitive to testosterone suppression.
Androgen Antagonists↗
Analytic validation of the enzyme multiplied immunoassay technique for the determination of mycophenolic acid in plasma from renal transplant recipients compared with a high-performance liquid chromatographic assay.
The analysis of mycophenolic acid (MPA) has proved a valuable adjunct to the clinical care of organ transplant recipients. The analytic validation of the enzyme multiplied immunoassay technique (EMIT) for the determination of MPA in plasma is described. The EMIT MPA standard curve was 0 to 15.0 microg/mL, and curve storage was maintained for 4 weeks. The MPA EMIT assay proved reliable and reproducible, as shown by the intra-assay and interassay coefficients of variation (1.58-3.68% and 1.23-7.57%, respectively). Excellent linear correlation ( r = 0.999) was observed for dilution linearity. The sensitivity of the assay was 0.01 microg/mL. Recoveries of 99.4% to 104.2% were obtained by spiking aliquots of three controls of known MPA concentrations with MPA. No interference was observed for endogenous substances and coadministered immunosuppressant drugs, and no cross-reactivity from the major metabolite MPA glucuronide was found. The high-performance liquid chromatography (HPLC) assay used protein precipitation and C18 ion-pair chromatography with ultraviolet detection at 304 nm. Plasma concentrations of MPA were measured using EMIT and HPLC. A linear relationship was observed between EMIT and HPLC (EMIT = 1.091 x HPLC - 0.089; r 2 = 0.990, n = 129). These results indicate that EMIT is a simple, rapid, and sensitive assay method for the measurement of MPA in plasma.
Chromatography, High Pressure Liquid↗
Enhanced vascular permeability in solid tumor involving peroxynitrite and matrix metalloproteinases.
Peroxynitrite (ONOO(-)), which is generated from nitric oxide (NO) and superoxide anion (O(2)(.-)) under pathological conditions, plays an important role in pathophysiological processes. Activation of matrix metalloproteinases (MMPs) contributes to tumor angiogenesis and metastasis. NO mediates the enhanced vascular permeability and retention (EPR) effect in solid tumors, and ONOO(-)activates proMMP to MMP in vitro. In this study, we examined the role of ONOO(-)in the EPR effect in solid tumors and normal tissues as related to MMP activation. Authentic ONOO(-), at 50 nmol or higher concentrations, induced the enhanced vascular permeability in normal dorsal skin of mice. ONOO(-)scavengers ebselen and uric acid significantly suppressed the EPR effect in mouse sarcoma 180 (S-180) tumors. Indirect evidence for formation of ONOO(-)in S-180 and mouse colon adenocarcinoma (C-38) tumors included strong immunostaining for nitrotyrosine in the tumor tissue, predominantly surrounding the tumor vessels. MMP inhibitor BE16627B (66.6 mg / kg i.v., given 2 times) or SI-27 (10 mg / kg i.p., given 2 times) significantly suppressed the ONOO(-)-induced EPR effect in S-180 tumors and in normal skin. Soybean trypsin inhibitor (Kunitz type), broad-spectrum proteinase inhibitor ovomacroglobulin, and bradykinin receptor antagonist HOE 140 also significantly suppressed the ONOO(-)-induced EPR effect in normal skin tissues. These data suggest that ONOO(-)may be involved in and promote the EPR effect in tumors, which could be mediated partly through activation of MMPs and a subsequent proteinase cascade to generate potent vasoactive mediators such as bradykinin.
Adenocarcinoma↗