Increased levels of serum matrix metalloproteinase-3 in haemodialysis patients with dialysis-related amyloidosis.

BACKGROUND: It is recognized that matrix metalloproteinase-3 (MMP-3) is abundantly expressed in active rheumatoid synovium, and that serum level of MMP-3 is a useful marker for diagnosis of rheumatoid arthritis and for evaluation of prognosis in joint destruction. Little is known about serum MMP-3 levels in haemodialysis (HD) patients, and thus, the association between serum MMP-3 and dialysis-related amyloidosis (DRA) has yet to be elucidated. METHODS: Serum levels of MMP-3 were measured by enzyme immunoassay in 150 HD patients, 90 without DRA and 60 with DRA, before HD. Simple regression analysis was performed to investigate the relationship between serum level of MMP-3 and clinical parameters, including age, HD duration, C-reactive protein and beta2 microglobulin (BMG). RESULTS: Serum levels of MMP-3 were significantly higher in HD patients with DRA than in HD patients without DRA (258.2 +/- 118.1 vs 201.5 +/- 98.4 pg/mL, P = 0.0017), and both levels were significantly higher than those of healthy subjects (45.6 +/- 13.4 pg/mL, P < 0.0001). Serum MMP-3 levels significantly correlated with serum levels of BMG (r = 0.197, P = 0.0164) and HD duration (r = 0.168, P = 0.0427). Moreover, serum MMP-3 levels significantly correlated with serum BMG levels in HD patients without DRA (r = 0.341, P = 0.0012), but not in HD patients with DRA. CONCLUSION: Our results suggest that matrix metalloproteinase activity increases in HD patients, which may be associated with BMG and DRA.

The effect of ultrapure dialysate on improving renal anemia.

BACKGROUND: Renal anemia is a very serious problem in hemodialysis patients. In this regard, the investigation was focused on whether ultrapure dialysate could improve renal anemia and the mechanism of renal anemia. METHODS: Ultrapure dialysate was used over a 2 years period for 61 patients on maintenance hemodialysis. During this period, the changes in renal anemia and red blood cell life span were investigated. The changes in the latter were evaluated using the creatine concentration in red blood cell. RESULTS: The hemoglobin concentration, RBC count, and hematocrit concentration before the use of the ultrapure dialysate were 9.1 +/- 0.2 g/dL, 309.9 +/- L7.2 x 10(4)/microL, and 28.8 +/- 0.6%, respectively. These values significantly increased to 10.2 +/- 0.1 g/dL, 349.7 +/- 5.6 x 10(4)/microL, and 32.6 +/- 0.3%, respectively, after 2 years of ultrapure dialysate use. The increase in reticulocyte count indicated enhanced erythropoiesis by ultrapure dialysate. The red blood cell life span evaluated by creatine concentration in red blood cell was also prolonged after the use of ultrapure dialysate. CONCLUSIONS: Ultrapure dialysate is considered to improve the renal anemia of dialysis patients by promoting erythropoiesis and prolonging red blood cell life span.

The prevalence of metabolic syndrome in Japanese renal transplant recipients.

BACKGROUND: The prevalence of metabolic syndrome (MS) after renal transplantation has yet to be elucidated. In the present study, we investigated the prevalence of MS in Japanese renal transplant recipients. METHODS: A cross-sectional study was conducted to determine the prevalence of MS in 101 renal transplant recipients at Osaka City University Hospital. The prevalence of MS was determined using the National Cholesterol Education Program-Adult Treatment Panel III (NCEP-ATPIII) criteria (modified and original) and the International Diabetes Federation (IDF) criteria. RESULTS: Using the modified (Japanese) NCEP criteria, a total of 24 out of 101 patients (23.8%) had MS including 21 out of 64 male patients (32.8%) and three out of 37 female patients (8.1%). Using the modified (Asian) NCEP criteria, MS was diagnosed in 23 patients (22.8%); 19 male (29.7%) and four female (10.8%). Using the original NCEP criteria, MS was diagnosed in 15 patients (14.9%); 12 male (18.8%) and three female (8.1%). Using the IDF criteria, MS was diagnosed in 16 patients (15.8%); 15 male (23.4%) and one female (2.7%). CONCLUSION: The prevalence of MS differed according to the NCEP criteria used, which had different cut-off points for waist circumference (14.9-23.8%). By the IDF criteria, which cites central obesity as an essential component, the prevalence of MS was slightly lower. Furthermore, in our study, MS was more prevalent in male renal transplant recipients.

Palliative treatment of bone metastases in hormone-refractory prostate cancer: effects of pamidronate on the carboxyterminal telopeptide of type-I collagen level in patients with increasing prostate-specific antigen levels.

PURPOSE: Bisphosphonates have been reported to be effective in reducing bone pain and skeletal-related events associated with bone metastases in hormone-refractory prostate cancer (HRPC). However, whether bone resorption is reduced primarily by these particular drugs is difficult to evaluate because patients with HRPC are usually treated with secondary or tertiary hormonal manipulations including second-line antiandrogens, high-dose diethylstilbestrol, or low-dose dexamethasone therapies, some of which may also be effective. Thus, we assessed changes in the level of the carboxyterminal telopeptide of type-I collagen (ICTP), a bone resorption marker, before and after pamidronate administration in HRPC patients with increasing prostate-specific antigen (PSA) levels. PATIENTS AND METHODS: Twenty-one HRPC patients with bone metastases and increasing PSA levels were intravenously treated with pamidronate at a dose of 30 mg either every 2 or every 4 weeks. Pamidronate administration was started immediately after confirmation of three consecutive increases in the PSA level. RESULTS: In 14 patients (67%), the ICTP levels decreased after the administration of pamidronate, despite increasing PSA levels. In 7 of these cases, the ICTP levels were lower than those recorded for 6 months or longer before the start of pamidronate administration. The characteristics of the responders were compared with those of the non-responders. CONCLUSION: In 67% of the HRPC patients with increasing PSA levels, pamidronate reduced the accelerated turnover of bone metabolism caused by metastases of prostate cancer.

Significance of target cell infection and natural killer cells in the anti-tumor effects of bacillus Calmette-Guerin in murine bladder cancer.

Although intravesical instillation of bacillus Calmette-Guerin (BCG) is a clinically well-recognized therapy for bladder carcinoma in situ and recurrence prophylaxis, these mechanisms have not been fully understood. We studied the effects of BCG infection (Connaught strain) on target cancer cells and host immune systems in murine bladder cancer. The bladder cancer cell line, MB49, was used in C57/BL6 mice in vivo and in vitro. In vitro cytotoxicities against the cancer cell line were measured by 24-h 51Cr release assay. For effector cells, spleen mononuclear cells were obtained from mice injected intraperitoneally with BCG or BCG-infected irradiated MB49 cells. Although BCG infection of cancer cells did not affect the proliferation speed in vitro, the mice injected subcutaneously with BCG-infected MB49 cells survived significantly longer than those given untreated cancer cells. The mice surviving without tumor growth after injection of BCG-infected cancer cells could not reject a second injection of intact MB49 cells. In vitro cytotoxicity was enhanced by BCG infection of target cancer cells, but not by immunizing the mice with BCG from which effector cells were obtained. Moreover, cytotoxicity disappeared by depleting natural killer (NK) cells from effector cells. Although in vitro cytotoxicity was increased by immunizing the mice with BCG-infected irradiated MB49 cells, survival did not improve in these mice. These results suggest that a major part of BCG's anti-tumor effects can be attributed to the elimination of BCG-infected cancer cells by NK cells.

[The effect of bisphosphonates on bone metastasis of hormone-refractory prostate cancer].

Approximately 70% of patients with advanced prostate cancer have bone metastases, which are associated with considerable skeletal morbidity, accompanied by severe bone pain that requires narcotics or palliative radiation therapy, pathological fractures, spinal cord compression and hypercalcemia of malignancy (HCM), which consequentiy lower the patient's quality of life. Bisphosphonates, potent inhibitors of osteoclast activity and survival, therefore inhibiting osteoclast-mediated bone absorption, transiently palliative bone pain and decrease analgesic usage in patients who have hormone-refractory prostate cancer (HRPC) with bone metastases. Recently, a randomized controlled trial showed that a third-generation bisphosphonate, zoledronic acid reduced bone pain and skeletal-related events (SREs). In this manuscript, we reviwed the efficacy of bisphosphonates in HRPC with bone metastases from several clinical studies and discuss treatment of advanced prostate cancer with bisphosphonates.

Silent cerebral infarction predicts vascular events in hemodialysis patients.

BACKGROUND: Cardiovascular disease is the leading cause of death in hemodialysis (HD) patients. We have previously reported a higher incidence of silent cerebral infarction (SCI) in HD patients compared with the control group using MRI studies. In the present study, we examined whether or not SCI could predict vascular events in HD patients. METHODS: Cranial magnetic resonance imaging (MRI) was performed on 119 HD patients without symptomatic cerebrovascular disease. SCI was detected by MRI, and the patients were prospectively followed up. The end points of the study were the incidence of major events related to vascular events (cerebral events, cardiac events, and sudden deaths). We investigated the prognostic role of SCI in cerebral, cardiac, and vascular events by using the Kaplan-Meier method and Cox proportional hazards analysis. RESULTS: The prevalence of SCI was 49.6% in HD patients. During a follow-up period of maximum 60 months, vascular events, which included 13 cerebral events, 5 cardiac events, and 3 sudden deaths, occurred in 21 patients. The presence of SCI was predictive for a higher cerebral and vascular morbidity compared to the absence of SCI [18.6% (N= 11) vs. 3.3% (N= 2), P= 0.0169, and 30.5% (N= 18) vs. 5.0% (N= 3), P= 0.0006, respectively]. By multivariate Cox proportional hazards analysis, SCI remained a powerful independent predictor of cerebral and vascular events (hazard ratio for cerebral events 7.33, 95% CI 1.27-42.25: for vascular events 4.48, 95% CI 1.09-18.41). CONCLUSION: The findings of the present study indicate that the presence of SCI is a new risk factor for vascular events in HD patients.

Influence of glucose and inflammatory cytokines on TGF-beta1 and CTGF mRNA expressions in human peritoneal mesothelial cells.

Peritoneal fibrosis is a major complication of long-term continuous ambulatory peritoneal dialysis (CAPD) treatment. Transforming growth factor-beta (TGF-beta) has been reported to play an important role in the fibrosis of various tissues by stimulating connective tissue growth factor (CTGF) expression. In order to elucidate the mechanism of CAPD-related peritoneal fibrosis, we studied the influence of high glucose concentrations and inflammatory cytokines on mRNA expressions of TGF-beta and CTGF in cultured human peritoneal mesothelial cells (HPMC). HPMC were isolated from normal omentum and cultured with 0.5 or 1.0% glucose or mannitol for 7 days. TGF-beta1 and CTGF mRNA were quantified by one step real-time reverse transcription-polymerase chain reaction (real-time RT-PCR). TGF-beta1 and CTGF mRNA expression levels were significantly increased (p<0.05) by glucose in a dose-dependent manner, but not by mannitol. The expression levels were correlated between TGF-beta1 and CTGF. Effects of inflammatory cytokines were also examined by adding tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) or interleukin-6 (IL-6) to the medium at 0.1 ng/ml for 2 days. TGF-beta1 expression tended to be increased by TNF-alpha and IL-6. On the other hand, CTGF expression was significantly decreased (p<0.01) by IL-1 but not changed by TNF-alpha or IL-6. These results suggest that high glucose concentration may play a central role in peritoneal fibrosis. Responses of TGF-beta1 and CTGF to inflammatory cytokines were not necessarily identical, suggesting that CTGF may be a better therapeutic target for peritoneal fibrosis than TGF-beta1.

[Neuroendocrine differentiation in adenocarcinoma of prostate during combined androgen blockade therapy: a case report].

Prostatic neuroendocrine (NE) carcinoma is a rare disease with a poor prognosis because of its rapid progression and the androgen-independent characteristic, for which no successful therapy is available presently. We report a case of NE differentiated prostate cancer, which was diagnosed as adenocarcinoma initially and progressed with NE differentiation during the combined androgen blockade therapy.

Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon-alpha in renal cell carcinoma cell lines.

OBJECTIVES: Although interferon-alpha (IFNalpha) and interleukin-2 are used in the treatment of advanced renal cell carcinoma (RCC), the rate of efficacy is about 15% and not satisfactory. Therefore, a more effective treatment is being investigated. In this study, we examined the combined effects of IFNalpha and 5-fluorouracil (5-FU) as well as 5-deoxy-5-fluorouridine (5'-DFUR), a produrg of 5-FU, in vitro. MATERIALS AND METHODS: Four RCC cell lines (OCUU1, OCUU3, OCUU4, OCUU5) were established at our laboratory from RCC patients. OS-RC-2, RCC10RGB, TUHR14TKB, TUHR4TKB, A498 and Caki-1 were obtained from a commercial source. The sensitivity of the 10 RCC cell lines to 5-FU and 5'-DFUR was evaluated using MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into FdUMP, was estimated by ELISA. RESULTS: When the 10 RCC cell lines were divided into the low TP expression group and high TP expression group at 2.0 U/ml protein, TP expression was not enhanced by IFNalpha in all 4 cell lines in the low TP expression group. Antitumor effects were not enhanced by IFNalpha in 3 out of 4 cell lines for 5-FU and in all 4 cell lines for 5'-DFUR. On the other hand, in the high TP expression group, TP expression was enhanced by IFNalpha in 5 out of 6 cell lines, and antitumor effects were enhanced by IFNalpha in 5 out of 6 cell lines for 5-FU and in all 6 cell lines for 5'-DFUR. In addition, there was a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR in all RCC cell lines. CONCLUSIONS: These results suggested that TP may be useful as a predictive factor in combination therapy with IFNalpha and 5-FU or 5'-DFUR, which may be a promising treatment for advanced RCC.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Effect of type I growth factor receptor tyrosine kinase inhibitors on phosphorylation and transactivation activity of the androgen receptor in prostate cancer cells: Ligand-independent activation of the N-terminal domain of the androgen receptor.

Although there have been several studies suggesting the involvement of growth factor receptor tyrosine kinases in ligand-independent activation of the androgen receptor (AR) and progression of prostate cancer, limited studies have been reported actually showing the enhancement of phosphorylation of the AR in vivo in response to growth factors or activation of their receptors in prostate cancer cells. In this study, we have demonstrated that overexpression of HER2/Neu enhanced in vivo phosphorylation of the AR and MAP kinase in DU-145 cells, and that the HER2/Neu inhibitor TAK165 reduced the HER2/Neu-enhanced phosphorylated AR and MAP kinase, indicating that the MAP kinase pathway seems to be involved in the phosphorylation of the AR by HER2/Neu. Both HER2/Neu inhibitor TAK165 and EGFR tyrosine kinase inhibitor gefitinib ('Iressa', ZD1839) successfully reduced the HER2/Neu-induced transactivation activity of the AR in PC-3 and DU-145 cells, suggesting that these inhibitors are possible therapeutic drugs for patients with hormone-refractory prostate cancer. The transactivation activity of the AF-1+DBD of the AR was enhanced by HER2/Neu overexpression while that of the AF-2+DBD was not, demonstrating that the enhancement of the AR activity by HER2/Neu was mainly mediated through the AF-1 of the AR.

Kidney transplants from living related donors having double inferior vena cava.

We performed four living related kidney transplantations from donors with double inferior vena cava (D-IVC), in which the left kidney was selected in two cases and the right in two cases. By dissecting the right internal iliac vein and isolating the right external iliac vein, the surgical procedure of the recipient side was so devised as to avoid any complications. In one patient, the surgical procedure of the donor side was modified to extend the donor left renal vein by anastomosis of part of the donor IVC to the renal vein. In the other case, no special treatment was necessary due to the patient's slender physique. In all four cases, transplants were successfully performed. The following conclusions can be made from these results: If the donor has D-IVC, it is essential to carefully conduct pre-operative examinations including angiography and venography to investigate other possible anomalies and blood flow of the renal vein. In addition, the graft must be carefully selected so that it is not disadvantageous to the donor. If there is no disadvantage to the donor as to which kidney is selected, the kidney with the longer renal vein should be transplanted.

Antitumor effects of lipoxygenase inhibitors on murine bladder cancer cell line (MBT-2).

Arachidonic acid (AA) metabolites derived from both the lipoxygenase (LOX) and cyclooxygenase (COX) pathways transduce a variety of signals related to cell growth. Selective blockade of the different metabolic pathways of AA (using a general LOX inhibitor NDGA, a 5-LOX inhibitor AA861, a 12-LOX inhibitor baicalein and a general COX inhibitor ibuprofen) revealed that murine bladder cancer cell line (MBT-2) cell proliferation was inhibited by the LOX inhibitors concentration-dependently, but not by the COX inhibitor. Among the LOX inhibitors, baicalein showed the strongest inhibition and induced apoptosis of MBT-2. Proliferation of MBT-2 was also significantly inhibited by 12-LOX antisense oligonucleotides. In an in vivo experiment, the antitumor effects of baicalein administration on C3H/HeN mice implanted with MBT-2 were recognized. These results suggested that LOX inhibition may be significant in the treatment of bladder cancer.

Biological reactions resulting from endotoxin adsorbed on dialysis membrane: an in vitro study.

Some types of dialysis membrane are known to adsorb endotoxin (ET). It is suggested that the biocompatibility of dialysis membrane is enhanced by adsorption and inhibition of ET. This study attempts to clarify the membrane-mediated biological reaction of the ET that is adsorbed to a dialysis membrane. After a dialysis circuit was prepared, contaminated dialysate was introduced on the dialysate side of a polyether polymer alloy (PEPA) membrane that adsorbs ET while saline solution or blood were introduced on the blood side, and the difference in ET adsorption between the two set-ups was measured. Further, the side filled with blood was left standing for 2 h, after which the changes in the amount of interleukin 1 receptor antagonist (IL-1Ra) produced from the whole blood were also assayed. Significantly more ET was adsorbed to the dialysis membrane when blood rather than saline was on the other side. In addition, the IL-1Ra production from the dialysis membrane that adsorbed ET was significantly higher. The ET adsorbed to the dialysis membrane may influence a living body even if it does not pass through the membrane. Accordingly, it is difficult to assume that the adsorption of ET to the membrane enhances its biocompatibility.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Endotoxin adsorption of various dialysis membranes: in vitro study.

Endotoxin (ET) in the dialysate is known to adsorb on dialysis membranes made of polyether polymer alloy (PEPA) and polymethylmethacrylate (PMMA). In the present study, we investigated the adsorption of ET on dialysis membranes with a focus on PEPA membranes and polysulfone (PS) membranes that are extensively used in artificial kidneys or as ET-removal filters. In the case of PEPA, the compounding of polyvinylpyrrolidone (PVP) was changed, and both a hydrophobic version and a hydrophilic version were used on the blood side. For the PS dialysis, commercial membranes (APS (Asahi), BSP (Toray), PSN (Fresenius), CLPS (Terumo)) were used. Adsorption was evaluated by exposing both sides of the membrane after it had been primed with physiological saline: the ET concentration on the blood side and dialysate side of the dialysis membrane was monitored during the 240 min from the start of the exposure. When the PEPA membrane was investigated, ET was significantly adsorbed on the hydrophobic version. For PS membranes, ET was adsorbed on the blood side or on both the blood and dialysate sides, depending on the membrane. PS dialysis membranes can adsorb ET but the power and site of adsorption are different even between membranes made of the same material. In addition to electrostatic action attributable to the compounding of hydrophilic agent PVP on the dialysis membrane, the distribution of PVP that was compounded and the potential of the membrane itself are considered to cause differences in adsorption.

Reduced-intensity stem cell transplantation in two cases of metastatic renal cell carcinoma.

Allogeneic hematopoietic stem-cell transplantation can induce curative graft-versus-leukemia reactions in patients with hematological malignancies. There is also evidence of such an effect in patients with solid tumors. We report two patients with metastatic renal cell carcinoma who underwent RIST. In both patients, disease progression was observed 6 months after transplantation. However, one patient had transient symptoms of tumor progression after the occurrence of acute graft-versus-host disease, consistent with graft-versus-tumor effects.

Role of tumor-associated macrophages in renal cell carcinoma.

We investigated the biologic meaning of tumor-associated macrophages (TAM) in renal cell carcinoma (RCC). The study group comprised of 83 patients with RCC. TAM was isolated by plastic adherence following enzymatic digestion of surgically removed tumor tissues. In some of the patients, monocytes were also isolated from peripheral blood mononuclear cells by plastic adherence. When TAM and monocytes were compared in the same patients, TAM produced interleukin 6 (IL-6), tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) without lipopolysaccharide (LPS) stimulation, while monocytes hardly produced IL-6, TNFalpha and IL-1beta without LPS stimulation. However, with LPS stimulation, monocytes produced more IL-6, TNFalpha and IL-1beta than TAM. In stage T1 RCC patients, there was a significant positive correlation between TNFalpha production of TAM and tumor size. In order to investigate the effects of TAM on cancer cells, TAM was co-cultured with A498, K562 and in some cases, with short-term established RCC lines for 96 h. As a result, TAM largely enhanced cell proliferation. These results suggested that TAM may play an important role in certain steps of tumor progression.