Role of thymidine phosphorylase in biomodulation of fluoropyrimidines.

Thymidine phosphorylase (TP) is a key enzyme in the activating pathway of 5'DFUR and capecitabine. On the other hand, TP is identical to platelet-derived endothelial cell growth factor (PD-ECGF) which is known to be an angiogenic factor. Recent studies show TP expression is increased in various malignancies compared with the surrounding normal tissues. These reports demonstrate that elevated TP expression indicates a predisposition for aggressive disease and/or poor prognosis. Therefore, it is a reasonable strategy to target TP in cancer treatment by using fluoropyrimidines including 5-fluorouracil (5FU), 5'DFUR and capecitabine. TP-mediated biomodulation of fluoropyrimidines to enhance their anti-tumor effects has been investigated. TP up-regulators including cytokines, anti-tumor drugs and X-ray irradiation significantly increase cytotoxicity of fluoropyrimidines. Also, transfection of TP cDNA significantly enhances cytotoxicity of fluoropyrimidines. Biomodulation of fluoropyrimidines is clinically successful in treating some malignancies. We report a review on roles of TP in biomodulation of fluoropyrimidines.

Isolation of an androgen-inducible novel lipocalin gene, Arg1, from androgen-dependent mouse mammary Shionogi carcinoma cells.

Here we report isolation of an androgen-regulated novel gene from an androgen-dependent mouse mammary Shionogi carcinoma SC-3 cell line. Using a polymerase chain reaction-based subtraction method and Northern blotting analysis, we isolated four androgen-inducible genes from SC-3 cells. Nucleotide sequencings identified three of the genes as cyclin D1, beta-catenin, and fatty acid synthase, respectively, but the fourth, a gene tentatively named as Arg1 (androgen-regulated gene 1), remained undefined. The cloned 2.0-kb sized Arg1 cDNA encoded 414 amino acid sequences. The deduced amino acid sequences, sharing about 30% homology with cathepsin family members at a protein level, had relatively conserved residues around the three proteinase active sites reported earlier. In Northern blotting, Arg1 mRNA was found in kidney, heart, lung, and to a lesser degree, in spleen and liver. Its transcripts were also detected in male reproductive organs on RT-PCR. In addition, its expression levels in prostate were markedly reduced after castration. Unexpectedly, Arg1-expressing COS1 cells showed no significant proteinase activity to various synthesized substrates under neutral or acidic conditions in this study. This might have been due to the replacement of the cysteinyl active site for proteinase to serine residue in the Arg1 amino acid sequences. Given that Arg1 also contains a lipocaline signature known as a binding motif for small hydrophobic molecules at the center of its amino acid sequences, Arg1 is a lipocalin family gene regulated by androgens in prostate and Shionogi carcinoma cells.

Predominant expression of fibroblast growth factor (FGF) 8, FGF4, and FGF receptor 1 in nonseminomatous and highly proliferative components of testicular germ cell tumors.

Nonseminomatous components within testicular germ cell tumors affect patient prognosis to varying degrees. These components are well known to mimic early embryonic totipotential tissues. Prompted by the recent observation that fibroblast growth factor (FGF) 8, FGF4, and FGF receptor (FGFR) 1 are required for the growth of early postimplantational embryonic tissues, we investigated the expressions of FGF8, FGF4, and FGFRI in surgically resected specimens of primary testicular germ cell tumors using an immunohistochemical method. All cases of embryonal carcinoma (14 cases), yolk sac tumor (3 cases), and choriocarcinoma (3 cases) showed positive immunostaining for FGF8, FGF4, and FGFR1. In contrast, out of 13 cases of seminoma, immunostaining was negative for FGF8, FGF4, and FGFR1 in 8 cases (61.5%), 6 cases (46.1%), and 7 cases (53.8%), respectively. In 7 cases of mature and immature teratoma, most areas showed negative immunostaining. In addition, the Ki-67 labeling index showed extremely high mitogenic activity in embryonal carcinoma, yolk sac tumor, and choriocarcinoma, which are precisely the carcinomas with the highest expressions of FGF8, FGF4, and FGFR1. It is in keeping with the immunohistochemical result that murine teratocarcinoma P19 cells were shown to express FGF8, FGF4, and FGFRI only under undifferentiated growth conditions. Taken together, these findings confirm the involvement of FGF8, FGF4, and FGFR1 in highly proliferative conditions of nonseminomatous germ cell tumors.

Use of a sponge stopper for placement of urethrovesical anastomotic sutures after radical prostatectomy or enterocystoplasty.

Vesicourethral anastomosis during radical retropubic prostatectomy or enterocystoplasty is often difficult because of poor visualization and collapse of the urethral stump. We have attempted to use a sponge stopper to facilitate urethral suture placement for vesicourethral anastomosis. The technique for use of a sponge stopper is described.

Biomodulation of 5-fluorouracil by interferon-alpha in human renal carcinoma cells: relationship to the expression of thymidine phosphorylase.

PURPOSE: To provide the basis for improved therapeutic benefit in combination chemotherapy with interferon (IFN) and 5-fluorouracil (5-FU), we investigated the modulatory actions of human recombinant IFN alfa-2a on 5-FU in five renal cell carcinoma (RCC) cell lines in vitro, in particular focusing on thymidine phosphorylase (TP) expression. METHODS: The sensitivity of RCC cell lines to the drugs was evaluated using an AlamarBlue assay. An enzyme-linked immunosorbent assay (ELISA) was used to determine TP expression. RESULTS: IFN-alpha enhanced the sensitivity of three of five RCC cell lines to 5-FU in a dose- and schedule-dependent manner. When IFN-alpha was given prior to 5-FU, sensitivity to 5-FU was significantly higher than when IFN-alpha was given simultaneously (P < 0.05). IFN-alpha enhanced TP expression in a dose-dependent manner in three of five RCC cell lines (P < 0.05). The relative IFN-alpha-induced increase in sensitivity to 5-FU correlated with the relative IFN-alpha-induced increase in TP expression (P < 0.05). In addition, two of three RCC cell lines with more than a twofold increase in sensitivity to 5-FU induced by IFN-alpha showed a higher TP expression without IFN-alpha stimulation. CONCLUSIONS: These results suggest that IFN-alpha upregulates TP expression and modulates 5-FU anabolism thus enhancing 5-FU cytotoxicity in a dose- and schedule-dependent manner in some RCC cells. The results imply that TP expression measurement in RCC could identify subgroups of metastatic RCC that may respond to IFN-alpha/5-FU combination therapy, and sequential administration of IFN-alpha followed by 5-FU may be beneficial in such cases.

Cisplatin-induced vomiting depends on circadian timing.

We examined whether the clock time of cisplatin plus antiemetic and diuretic administration affects the amount of cisplatin-associated emesis and severity of renal toxicity. We treated 22 patients with urogenital cancer with two courses of chemotherapy containing 70 mg/m2 of cisplatin. Cisplatin together with furosemide was administered in the morning (05:00) or evening (17:00) during two courses 1 month apart in a crossover fashion. Ondansetron was given either before or after cisplatin to control nausea and vomiting. The number of vomiting episodes, serum creatinine, serum urea nitrogen (BUN), creatinine clearance, and urinary beta-N-acetyl glucosamidase (NAG) concentration were evaluated before and after each treatment course. Regardless of the timing of ondansetron, morning compared to evening cisplatin was always associated with greater vomiting in the first treatment course. However, prophylactic administration of ondansetron markedly diminished the impact of the clock time of cisplatin administration. Serum creatinine transiently decreased rather than increased 14 days after cisplatin and furosemide administration, while NAG excretion increased 3 days after cisplatin and furosemide administration. In the first course, serum creatinine levels were similar regardless of the clock time of cisplatin and furosemide administration. However, in the second course, serum creatinine rose in patients given evening cisplatin and furosemide, while it remained unchanged in those given morning cisplatin and furosemide. Moreover, the first course morning cisplatin and furosemide treatment was associated with less change in NAG excretion (less kidney toxicity) than the first course of evening cisplatin and furosemide treatment. The second course evening cisplatin and furosemide treatment was associated with an increase in NAG excretion compared to the first course of treatment, while morning cisplatin and furosemide treatment in the second course showed less change in NAG excretion compared to the first course. The clock time of cisplatin administration had an impact on the frequency of emesis. Prophylactic ondansetron, however, diminished the time-of-day dependency of cisplatin-induced vomiting. Administration of cisplatin and furosemide in the morning rather than evening appears to cause less renal damage, and this damage may be further reduced with aggressive hydration and routine administration of furosemide.

Effects of dosing time and schedule on cisplatin-induced nephrotoxicity in rats.

Renal dysfunction induced by a single injection of cisplatin depends on the timing of the dose. However, the effects of repeated administration of cisplatin on time-dependent toxicity have not been evaluated despite the fact that in clinical practice high doses are repeatedly injected at intervals or low doses are administered daily. We studied chrononephrotoxicity in rats after weekly or daily cisplatin injections. Weekly high doses (5 mg kg(-1)) or daily low doses (1.2 mg kg(-1)) of cisplatin were injected at four time points (3, 9, 15 and 21 h after the light was turned on (HALO)) for 3 weeks. Changes in body weight after weekly cisplatin administration were independent of the timing of the doses. In the group that received daily cisplatin, the loss in body weight in the 3 HALO group was smaller than in animals receiving injections at 15 and 21 HALO (P < 0.05 and 0.001, respectively). The blood urea nitrogen and serum creatinine levels in the control rats showed a significant circadian change (peak at 15 HALO and trough at 9 HALO), but these parameters were markedly elevated in both trials and their circadian variations were disturbed. In conclusion, cisplatin-induced nephrotoxicity was the lowest at 3HALO compared with other time points of both dose regimens.

[Serum CA19-9 levels in testicular germ cell tumor patients].

This study was designed to examine whether measurement of serum CA19-9 was useful in testicular germ cell tumor patients. We analyzed the clinical courses of 55 testicular germ cell tumor cases diagnosed after high orchiectomy. The patients in this study consisted of 33 seminomas and 22 non-seminomatous germ cell tumors (NSGCT), and their mean age was 32.7 +/- 12.7 years (mean +/- SD). The mean follow-up period after the operation was 33.7 months. The positive rate of the pre-treatment serum CA19-9 level was 16.4% (3.0% in seminomas versus 36.4% in NSGCT, p = 0.0017). The pre-treatment serum CA19-9 levels in NSGCT patients were significantly higher than those in seminoma patients (46.6 +/- 50.0 U/ml versus 10.6 +/- 9.6 U/ml, p = 0.0008). We divided the patients into two groups according to the detailed histological types, and found that the serum CA19-9 levels in the patients with embryonal carcinoma (EC) were significantly higher than in those without EC (p = 0.0160), and the levels in those with yolk sac tumor (YS) were higher than in those without YS (p = 0.0099). Moreover, the levels in those with either EC or YS were significantly higher than in those with neither EC nor YS (p = 0.0004). In 9 patients with a high serum pre-treatment CA19-9 level, the serum CA19-9 level was useful as a monitoring marker through the treatment or tumor progression. On the other hand, the pre-treatment serum CA19-9 level did not correlate with the clinical stage or prognosis. In conclusion, the phenomenon that the serum levels of CA19-9 increase in testicular germ cell tumor patients is not extremely rare, and in NSGCT, especially in EC or YS, the serum CA19-9 can be a useful tumor marker.

[Clinical usefulness of cross-linked N-telopeptide of type I collagen (NTx) as a bone metastatic marker in patients with prostate cancer--comparison with serum PICP, PINP and ICTP].

Type I collagen cross-linked N-telopeptide (NTx) in urine, the degraded form of type I collagen cross-linked in bone, has been evaluated as a marker of bone resorption. In this study, the clinical usefulness of NTx as a marker of bone metastasis of prostate cancer was compared with that the carboxyterminal propeptide of type I procollagen (PICP), the aminoterminal propeptide of type I procollagen (PINP), and the pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP) in serum. We assessed 37 cases of prostatic cancer in which the diagnosis had been confirmed pathologically. The patients were 15 patients with prostatic cancer with bone metastasis (before treatment or during a relapse) (Group 1); 11 patients, with bone metastasis, but for whom treatment was effective and condition had stabilized (Group 2); and 11 patients, with localized prostatic cancer and no evidence of bone metastasis (Group 3). The serum PICP, PINP, and ICTP levels and concentration of NTx in urine were compared among the three groups with the Mann-Whitney U test, with p values less than 0.05 considered significant. Urine NTx concentrations in Groups 1, 2 and 3 were 539.3 +/- 202.9, 160.6 +/- 97.6 and 48.6 +/- 7.6 nMBCE/mMCr, respectively. The differences between the Group 1 and Group 2 and between Group 1 and Group 3 were significant (p < 0.01 and p < 0.001). The differences between Group 1 and Group 3 and between Group 2 and Group 3 were significant for serum PICP, PINP and ICTP concentrations (p < 0.05). The correlation coefficient between urine NTx and each serum bone metabolic marker was 0.8 for PICP, 0.4 for PINP and 0.5 for ICTP. These bone metabolic markers are promising clinical markers of bone metastatic and may be useful for prediction of therapeutic efficacy and recurrence in bone and quantification of the extent of bone metastates.

[Clinical effects of beta-sitosterol (phytosterol) on benign prostatic hyperplasia: preliminary study].

Phytosterol derived from plants has long been used for the medical treatment of benign prostatic hyperplasia (BPH) in Europe but not in Japan. The efficacy of phytosterol was evaluated in patients with manifestations of urinary outlet obstruction caused by BPH. Phytosterol containing 180 mg of sitosterol per day was given to 12 patients with BPH in two or three divided doses for three months. The symptoms were assessed monthly using the International Prostate Symptom Score (IPSS) and quality-of-life (QOL) score while the objective findings including the urinary flow, prostatic volume, and residual urine volume were assessed after three months of treatment of BPH. The IPSS and QOL scores showed significant improvement (p < 0.05), and the peak flow rate and residual urine volume showed slight but not significant improvement.

[A case of giant small cell carcinoma of the prostate].

A 64-year-old man having backbone pain was referred to our institute, and laboratory examinations revealed that he was suffering from severe renal failure. Abdomino-pelvic CT revealed bilateral hydronephrosis, a giant prostate over 500 g in weight, and multiple para-aortic lymph node metastasis. Histological diagnosis of the prostate was small cell neuroendocrine carcinoma. Serum neuron-specific enolase (NSE) level was slightly high reflecting the positive immunohistochemical staining, but serum PSA remained within normal limits. The patient was treated with hormone-chemotherapy after successful treatment of the renal failure, but he died on the 74th hospital day. Small cell carcinoma is known to have a high rate of malignancy and metastasis from an early stage. Several giant prostatic tumors have been reported previously, but this case is considered to be the second gigantic small cell carcinoma in the Japanese literature.

[Estimation of the weddellite to whewellite ratio by infrared spectroscopy].

Powder samples of 56 calcium oxalate stones the contents of weddellite, whewellite and apatite of which had been determined by thermogravimetry (TG) were studied by infrared spectroscopy (IR). When the ratio of the weddellite content to the total of weddellite and whewellite (weddellite ratio) calculated using Oka's method on the infrared spectra was compared with that determined previously by TG, the correlation coefficient between these ratios was 0.734. For the 24 calcium oxalate stones the apatite content of which was less than 10%, the correlation coefficient between these ratios was 0.976, although the weddellite ratio calculated using Oka's method was significantly higher than that determined by TG (P less than 0.01). In an attempt to estimate the weddellite to whewellite ratio by IR regardless of the content of apatite, the ratio of the depth of the absorption band at 780 cm-1 (H780) to the depth of the absorption band at 1,320 cm-1 (H1320) was compared with the ratio of the whewellite content to the total of whewellite and weddellite contents determined by TG. The correlation coefficient between these ratios was 0.953. We conclude that the ratio of H780 to H1320 determined from the infrared spectra obtained from a 0.3 to 1 mg powder sample of calcium oxalate stone is useful in the estimation of the weddellite to whewellite ratio.

[A case of colo-vesico-vaginal fistula caused by sigmoid colon diverticulitis].

A 74-year-old woman was referred to our hospital with the chief complaints of pneumaturia, fecaluria and discharge of feces and urine from vagina. Fistulography on the vaginal side showed the presence of contrast medium both in the sigmoid colon and bladder. Colonoscopy revealed multiple diverticulosis of the sigmoid colon. Under diagnosis of colo-vesico-vaginal fistula due to sigmoid colon diverticulitis, a one-stage operation removing sigmoid colon, uterus-vaginal wall and urinary bladder wall including the fistula and careful reconstruction was performed. Postoperatively, urinary leakage from vagina in large amounts continued due to the recurrence of vesico-vaginal fistula. An attempt to use human fibrin glue in the recurrent fistula was successful, and the patient was asymptomatic at 21 months of follow-up. Colovesical fistula has been reported in about 10-20% of patients undergoing surgery for complicated diverticulitis, but a combined fistula is a rare condition. Furthermore, we recommend the use of human fibrin glue for a recurrent fistula.

[Clinical evaluation of urinary nuclear matrix protein 22 as a marker for bladder cancer].

The purpose of this study is to evaluate the clinical usefulness of urinary nuclear matrix protein 22 (NMP22) as a marker for bladder cancer. We examined the positive rates of NMP22 test, urinary cytology and bladder tumor antigen (BTA) test, and compared the positive rate of NMP22 test with that in urinary cytology and BTA test. Urine samples were obtained from 50 patients with histologically confirmed bladder cancer before the treatment. The samples were examined by NMP22 test, urinary cytology and BTA test. In 50 patients with bladder cancer, the overall positive rate was 40% for NMP22 test, 40% for urinary cytology, and 16% for BTA test. A combination of NMP22 test and urinary cytology showed a significantly higher positive rate (54%) as compared to NMP22 test or urinary cytology alone. When NMP22 test and urinary cytology were compared for tumor size, number, shape, stage and grade, NMP22 test showed a significant higher positive rate than urinary cytology in grade 1 bladder cancer. In conclusion, although NMP22 test and urinary cytology gave a similar positive rate, a combination of NMP22 test and urinary cytology is more useful than the NMP22 test or urinary cytology alone for monitoring of bladder cancer.

[Stage pT1 signet ring cell carcinoma of the urinary bladder: a case report].

A case of primary signet ring cell carcinoma of the urinary bladder is described. A 59-year-old man presented with microscopic hematuria, and cystoscopy revealed a white nonpapillary tumor. Histopathological examination of the resected tumor revealed signet ring cell carcinoma and transitional cell carcinoma. Histological depth of invasion was pT1. No adjuvant therapy was performed. Primary signet ring cell of the urinary bladder is a rare tumor with 37 cases reported to date in Japan. We investigated previously reported cases and discussed adjuvant therapies of superficial signet ring cell carcinoma of the urinary bladder.

[Gastrocystoplasty for severely contracted bladder following high dose pelvic irradiation: a case report].

A 24-year-old female who had received hysterectomy and adnexectomy of bilateral appendages for yolk sac tumor at the age of 12 years, followed by repeated surgery and pelvic irradiation over a total of 100 Gy for relapse of tumor, suffered from a severely contracted bladder and renal dysfunction of serum creatinine level over 2.0 mg/dl. The diagnosis was radiation-induced contracted bladder with renal dysfunction due to vesicoureteral reflux. Since the small intestine was not considered suitable after high dose irradiation, the stomach was used to reconstruct the bladder. The vesical pressure, which was 70 cmH2O at the capacity of 30 ml, was decreased to 22 cmH2O at the capacity of 100 ml, 5 weeks after surgery. The renal function was stable with a serum creatinine level below 1.4 mg/dl and the bladder capacity was 200 ml, 15 months after surgery. This method using the stomach was considered valuable for bladder reconstruction after large dose pelvic irradiation.

[The role of urine pH in the occurrence of phosphaturia].

The density of calcium phosphate and the pH were determined in 15 postprandial urine specimens with heavy precipitation of amorphous calcium phosphate, that is phosphaturia, collected from 5 patients with calcium urolithiasis (stone-formers) and 3 patients with no known urological disease (controls). Phosphaturia, not related to urinary tract infection or administration of alkalinizing agents, was found repeatedly at our outpatient clinic in these patients. The correlative relationship was not confirmed between the density of calcium phosphate and the pH. The concentration of calcium and phosphorus was also determined in 10 urine specimens with phosphaturia. The concentration of phosphorus was correlated significantly with the pH (r = -0.775, p less than 0.01), although the concentration of calcium was not correlated with the pH. The pH of 11 urine specimens with phosphaturia from the controls was 7.51 +/- 0.31 (mean +/- S.D.) and the pH of 18 urine specimens with phosphaturia from the stone-formers was 6.81 +/- 0.34. The pH was significantly lower in the urine specimens from the stone-formers than in those from the controls (p less than 0.01). We have noted that the occurrence of phosphaturia depends not only on the urinary pH but on the concentration of phosphorus. It is interesting that phosphaturia often occurs in urine specimens with a pH below 7 in stone-formers.

[Six cases of upper urinary tract diseases including tumors and inflammatory lesions which suggest the significance of urine cytology in preoperative diagnosis].

We present six cases of upper urinary tract diseases including tumors and inflammatory lesions in which the urine cytology rather than the radiological examinations was useful for their preoperative diagnoses. Three of the six cases had malignant diseases and the others had benign diseases. In all cases preoperative results of urine cytology were identical to histopathological findings of resected specimens; the cases with positive findings in urine cytology had ureter cancers and those with negative findings had benign diseases. Primary CIS of upper urinary tract was found in two of six cases, which is still uncommon in Japan. Since it is very difficult to make a preoperative diagnosis of primary CIS by radiological examinations, the present study showed that urine cytology is useful for its preoperative diagnosis. Recently endoscopic techniques for the diagnosis of upper urinary tract tumors are in clinical use. The ureteroscopic biopsy is recommended for the case in which the diagnosis using urine cytology is difficult.

[Squamous carcinoma with pseudosarcomatous stroma of the renal pelvis and ureter: a case report].

A case of unusual tumor of renal pelvis and ureter is reported. A 82-year-old man was admitted with the chief complaint of gross hematuria. He was diagnosed with bladder tumor and right renal pelvic and ureter tumor by bladder biopsy (TCC G2) and radiological examinations. Right nephroureterectomy was performed after transurethral resection of bladder tumor (TUR-Bt). Histological findings of renal pelvic and ureteral tumor were squamous carcinoma with pseudosarcomatous stroma (so-called carcinosarcoma). Tumor cells showed mostly a sarcomatoid spindle pattern and partly apparent SCC. He was discharged without adjuvant therapy and no recurrence was found for 7 months after operation. In our case, CA19-9 was useful as a tumor marker. Such disease has been confused with true carcinosarcoma or sarcomatoid carcinoma and should be distinguished from them. We reviewed 18 case reports of true carcinosarcoma or sarcomatoid carcinoma of the renal pelvis.

[A case of metastatic urinary bladder tumor from gastric carcinoma--especially mucohistochemical study].

We report a case of metastatic bladder tumor from gastric cancer, which was difficult to differentiate from urachal tumor preoperatively, especially computed tomographic scan, cystoscopy, and biopsy. A 51-year old man visited our hospital with the chief complaint of asymptomatic gross hematuria. He had received subtotal gastrectomy for gastric cancer two years earlier. Because the possibility of urachal tumor could not be excluded, en bloc segmental resection of the bladder, the urachus, and the umbilicus was carried out. This case was diagnosed as adenocarcinoma histologically, but it was difficult to determine whether the case was a metastatic bladder tumor from gastric cancer or urachal tumor by the routine staining method. High iron diamine-alcian blue and paradoxical concanavalin A (ConA) stainings were performed, on the surgical specimens of this case and other urachal tumor already diagnosed. In these cases, metastatic bladder tumor could be differentiated from an urachal one by mucohistochemically paradoxical Con A staining. Seventeen cases of metastatic bladder tumor from gastric cancer including our cases were collected from the Japanese literature and reviewed.