Cloning and expression of five myb-related genes from rice seed.

Three elements in the promoter of rice glutelin genes are important for their endosperm specific expression. One of these, an AACA motif, has been shown to be a negative regulator in non-seed tissues and has a similarity to the barley gibberellin responsive element recognized by MYB-like DNA binding proteins. A cDNA library constructed from immature rice seed was screened using two types of myb gene probes to isolate cDNA clones representing genes encoding MYB-like DNA binding proteins that may recognize the AACA motif in rice glutelin gene promoter. We obtained four cDNA clones encoding MYB-related proteins, Oryza sativa MYB (OSMYB) 1-4, using the maize C1 probe. Another myb-like clone, Osmyb5, was obtained by screening a rice seed cDNA library with probes designed to recognize the AACA-like binding domain in GAMYB and PHMYB3. RT-PCR was used to analyze Osmyb expression during rice seed development and their presence in other rice tissues, as it was not possible to detect these mRNAs by conventional Northern analysis. RT-PCR analysis showed that Osmyb2, Osmyb3 and Osmyb5 genes were expressed in all tissues examined. In seed, the mRNA levels of Osmyb1 and Osmyb4 genes reached a maximum at 14 days after flowering (DAF), suggesting that these genes may play a role in seed maturation. As Osmyb5 exhibits a high similarity to the regions in both GAMYB and PHMYB3, which can bind to the AACA motif, there is a possibility that the OSMYB5 protein may bind to the AACA motif of glutelin genes.

A 45-bp proximal region containing AACA and GCN4 motif is sufficient to confer endosperm-specific expression of the rice storage protein glutelin gene, GluA-3.

A 45-bp proximal region of the rice glutelin promoter (-104/-60) containing two putative cis-elements, the AACA motif and GCN4 motifs, was fused to a truncated CaMV 35S promoter (-90/+9; -90 delta 35S)/GUS. The 45-bp fragment specifically enhanced the promoter activity in endosperm tissue of transformed tobacco. A substitution mutation of the GCN4 motif reduced the promoter activity, whereas mutation of the AACA motif increased the activity in the embryo as well as in the endosperm. These results suggest that the GCN4 motif generally enhances the promoter activity but that the combination of the two motifs confers the endosperm specificity. Furthermore, the function of the two motifs was dependent on the orientation and/or distance from a G-box element in -90 delta 35S, suggesting that synergistic interaction between the factors that recognize those motifs and the G-box element is important for transcriptional regulation.

A case report of inflammatory pseudotumor of the urinary bladder.

A case of an inflammatory pseudotumor of the urinary bladder in a 46-yr-old man is presented. This rare, benign, and presumed non-neoplastic, reactive lesion must be differentiated from sarcomas of the urinary bladder. In the present case, we could demonstrate an inflammatory and reactive nature for the pseudotumor. Histologically, the presence of many Brunn's nests with infiltration of inflammatory cells and proliferation of capillaries in the myxoid stroma indicated a chronic inflammatory background for this lesion. It is apparent from morphology and immunohistochemistry findings that the proliferating spindle-shaped cell is of mesenchymal origin and not malignant in nature. These findings suggest that chronic inflammation can induce an overreaction of the bladder wall resembling tumor formation.

[Clinical experience with S-6436 in acute pyelonephritis (author's transl)].

S-6436 (a preparation consisting of 30% of non-enteric coated granules and 70% of enteric coated ones) was orally administered to the patients with acute pyelonephritis, and the following results were obtained: 1. Subjective symptoms: Very good in 13 cases (65%), good in 1, fair in 2 and poor in 4. 2. Urine finding: Very good in 13 cases (65%), good in 3 and poor in 4. 3. Disappearance of causative organisms: Organisms which were identified in the first examination were 17 strains of E. coli, one each of Pr. vulgalis, Klebsiella and Staph. epidermidis, 3 of Strept. faecalis, and 3 of mixed infections. Very good effect was observed in 16 cases, good in 1, fair in 1 and poor in 2. 4. Total evaluation: Very good in 13 cases (65%), good in 4 (20%) and poor in 3 (15%). 5. Slight discomfort in abdomen was observed in only one patient, but disappeared by administration of digestive. 6. S-6436 was given to the patients with acute pyelonephritis twice a day expecting that the drug has satisfactory clinical efficacy.

[Pharmacokinetic study of cefsulodin in patients with kidney dysfunction].

Seventeen patients with various grades of kidney function were injected each with cefsulodin (CFS) 1 g dissolved in 20 ml of saline. Serum and urine concentrations of CFS were determined using high performance liquid chromatography up to 24 hours after administration. Pharmacokinetic analysis was done using a two-compartment model. The results were discussed by comparing creatinine clearance (Ccr) values divided into 4 groups: greater than or equal to 70 ml/min (group I), 50-less than 70 ml/min (group II), 30-less than 50 ml/min (group III), and less than 30 ml/min (group IV). A delay in the disappearance of CFS from the blood was observed with a decrease in Ccr. Half-lives of CFS in blood (T 1/2 beta) were 1.03 hours (group I), 2.09 hours (group II), 3.44 hours (group III), and 4.52 hours (group IV). Serum clearance (Cls) and Ccr were found to be related with an equation: Cls = 1.60 x Ccr + 7.70. The correlation coefficient (r) was 0.881. Ccr and T 1/2 beta were found to be related with an equation: T 1/2 beta = 31.27 x Ccr-0.688, and the area under the curve (AUC) was found to be related to Ccr with AUC = 4,226 x Ccr-0.81. Urinary excretion rates up to 24 hours after administration were 84.4% (group I), 69.1% (group II), 67.5% (group III), and 56.5% (group IV). This means that CFS is excreted in urine with a relatively high recovery even in the case of low Ccr less than 30 ml/min.

[Pharmacokinetic study of cefmenoxime concerning its transfer into tissues].

Cefmenoxime (1 g) was intravenously administered by one shot to investigate its pharmacokinetic profile with respect to transfer into the kidney, vesical wall and prostatic adenoma. The concentration of cefmenoxime in the kidney reached a peak of 403 micrograms/g at 0.17 hour after administration. The biological half-life was 0.74 hour. In the vesical wall, the level of cefmenoxime reached a peak of 28 micrograms/g at 0.67 hour after administration. The half-life was 2.30 hours. The peak level of cefmenoxime in the prostatic adenoma was 22 micrograms/g at 0.39 hour after administration. The half-life was 2.90 hours. The concentrations of cefmenoxime in these urogenital tissues were higher than its MIC80 and MBC80 against various Gram-negative organisms.

[Morphological response of urinary bacteria to ceftibuten].

UNLABELLED: The morphological response of urinary Gram-negative bacteria to ceftibuten (CETB) was investigated in four patients with urinary tract infections (one patient: acute uncomplicated cystitis, three patients: chronic complicated urinary tract infection). The daily dose of CETB was 400 mg administered orally and the durations were 3 days for the acute uncomplicated cystitis case and 5 days for the chronic complicated urinary tract infection cases. In the four patients, changes in urinary CETB concentrations, viable bacterial counts, and morphology of the bacteria were studied after the initial administration. The urinary concentrations of CETB were 7.38-60.3 micrograms/ml at one hour after the first administration. The urinary viable cell counts were 1-5 x 10(7) cells/ml before administration, but they were reduced to 1-3 x 10(3) cells/ml at one hour after the first oral administration. Morphological changes of bacteria: Under phase contrast microscopy, filamentous debris and spherical bacteria with severe body damage were observed at 30 minutes after the first administration. By transmission electron microscopy, the cell wall of the filamentous bacteria showed a number of projections with formation of vacuolar structures in the space between the cell wall and the irregularly-shaped cytoplasm. According to the Japanese UTI criteria, the efficacy rate was 100% in the 4 patients. Neither side effect nor abnormal laboratory result was observed in any patient. CONCLUSION: The morphological changes of the bacteria suggested that CETB, in vivo, bounds not only penicillin-binding protein (PBP)-3 but also PBP-1.

[Clinical evaluation of serum ferritin levels in urologic cancer (II)].

By means of the RIA-Gnost Ferritin and Spac Ferritin kits, the serum ferritin levels of patients with some urological malignant diseases were quantitated; and, the clinical significance of these values as a tumor marker was discussed. The normal (reference) level in serum obtained by the RIA-Gnost kit was 142.5 +/- 67.5 ng/ml (n = 58) for healthy males and 49.6 +/- 28.8 ng/ml (n = 76) for healthy females. The assay values for 22 cases of bladder cancer by both kits were somewhat lower than the normal range, and the positive rates presented were around 30%, being in rough agreement with each other. The positive rate for 22 patients with prostatic cancer was 45.5% by either method. The assay values obtained by the RIA-Gnost kit was higher and differed significantly (p less than 0.1%) from those obtained on healthy males. Furthermore, the same tendency was found in 6 cases of renal cancer. The assay values of RIA-Gnost were significantly higher (p less than 0.1%) than normal; and, the positive rate was 100% for RIA-Gnost and 67% for Spac . In conclusion, serum ferritin determination especially that by the RIA-Gnost kit proved to be a useful and significant tumor marker for the detection of renal cancer.

[Our experience with combined intravesical instillation therapy of superficial bladder tumor using Aclacinomycin-A (ACM) and cytosine arabinoside (CA)].

Thirteen patients with recurrent superficial bladder tumors were treated by combined intravesical instillation of Aclacinomycin-A (ACM) and cytosine arabinoside (CA). Prophylactic effects of this combined instillation therapy were studied in 7 patients. A solution of 200 micrograms/ml of ACM and 600 micrograms/ml of CA was instilled into the bladder. The instillation aimed for treatment was carried out once a week until ten treatments had been given. Complete response was attained in 2 patients and partial response in 3 patients, but tumor size increased by more than 50% in 6 of the 13 patients. No change was observed in the remaining 2 patients. Recurrence of the tumors was observed in 3 of the 7 patients who were treated by this prophylactic combined instillation therapy. Local side effects such as bladder irritability were found in 2 of the 20 patients. No systemic side effects were noted in any patients. Although the side effects were reduced, we were not satisfied with the results of this therapy.

[Long-term treatment and clinical evaluation of enoxacin in complicated urinary tract infections].

Enoxacin (ENX) was used in the treatment of 16 patients with complicated urinary tract infections. ENX was orally administered 600 mg three times a day for 14--32 days. Clinical results evaluated according to the criteria of UTI committee were excellent in 6 cases, moderate in 3 cases and poor in 6 cases. The overall efficacy rate was 60.0%. Two moderate cases at 5 days showed excellent results at 14 days. Twelve out of 18 bacteriological strains were eradicated, with the elimination rate of 66.7%. As side effects, slight constipation occurred in one patient, and he was treated with purgatives. Photoallergic eruptions occurred in one of the 16 patients. He was treated with an antihistaminic agent and soon recovered.

[Experience using the laser rod in urological surgery].

The clinical use of a new surgical tool the "Laser Rod" is presented. This rod is made of ceramics and utilizes a neodymium-doped yttrium aluminum garnet (Nd-YAG) laser. With the Laser Rod, the surgeon can cut tissue by mechanical contact and still feel the resistance from the target tissue. In this study, this device has been used in various urological surgical procedures, such as nephrolithotomy, retropubic prostatectomy, ileocecal conduit diversion, urethroplasty and circumcision of the penis. Our results showed that laser surgery using this device, enables bloodless incision with minimal damage to adjacent tissue. In conclusion, the Laser Rod will probably be very useful in urological surgery, and its indications should be expanded through further study.

[Statistics on outpatients and inpatients at the urological department of Kosei Hospital 1975-1982].

Statistical observations of 19,678 outpatients and 2,601 inpatients at our department from 1975 to 1982 revealed the following results. The new cases in the outpatient clinic were in decreasing order nonspecific infections (50.8%), urogenital tumors (11.9%), urolithiasis (9.3%) and urogenital anomalies (8.0%). The inpatients according to the organ were in decreasing order kidney (29.6%), prostate (25.5%), bladder (13.4%) and ureter (11.9%). Operations were performed 2,414 times. Frequent operations were in decreasing order phimosectomy (360 times, 14.9%), TUR-Bt (284 times, 11.8%), transurethral cryosurgery of the prostate (215 times, 8.9%), vasectomy (190 times, 7.9%) and orchiopexy (152 times, 6.3%). More young persons comprised the outpatients of our hospital than at other facilities.

[Clinical evaluation of Claforan (CTX) in aged patients with chronic complicated urinary tract infections].

Claforan (CTX) was administered for 5 days at a daily dose of 4 g in two divided doses by intravenous injection to 18 in-patients, all of whom were over 70 years old and had chronic complicated urinary tract infections. The overall clinical efficacy of CTX was excellent in 4 cases, moderate in 8 cases and poor in 6 cases, the overall effectiveness rate being 66.7%. Twenty seven out of 34 organisms isolated from chronic complicated urinary tract infections before administration of CTX were eradicated after treatment. Both clinical findings and laboratory findings revealed no side effects. It is suggested that CTX is useful and a safe drug for elderly patients.

[Combination (RaHPP) of radiotherapy, hyperthermia and chemotherapy (peplomycin and picibanil) for bladder cancer].

Four patients with urinary bladder carcinoma were treated by combination therapy which consisted of hyperthermia vesical irrigation of two anticancer drugs (peplomycin and picibanil), intravesical instillation of those drugs and radiation. Following the therapeutic method we planned, 40 mg of peplomycin and 10 KE of picibanil in 1,500 ml of sterile distilled water was irrigated at 42 to 43 degrees C into the bladder for 3 hours; 40 mg of peplomycin and 10 KE of picibanil in 40 ml of sterile distilled water was instilled into the bladder; and, the focus was irradiated with 60Co to a focal dose of 200 rad 30 minutes later. This pattern of treatment was repeated once a week, 3 to 5 times in total. On the days this pattern was not taken, 5 KE of picibanil in 20 ml of sterile distilled water was instilled into the bladder cavity. Complete response was observed in one patient and partial response in 3 patients. The side effect was temporary irritable bladder symptom.

[Evaluation of the free nipple method in cutaneous ureterostomy].

Cutaneous ureterostomies, in which urostomas were made by a free nipple method, were performed on 14 ureters of 10 patients. The free nipple method was evaluated from the standing point of stoma care. Catheterless cutaneous ureterostomy was applied in 8 of the 10 patients. Six patients had to have catheters, and the catheter was not needed in 2 patients. The reasons for reindwelling the catheter in 6 patients were: 1) the urostoma had come to be at skin level by disturbance of blood supply for the ureter, and 2) urine puddled just on the urostoma and oozed out between the skin and Varicare flange. It may be concluded that the free nipple method is not satisfactory as urostoma in cutaneous ureterostomy for advancing the quality of life of the ostomate. A further study is in progress.

[Two cases of renal oncocytoma].

Two cases of renal oncocytoma are reported. These patients were a 67-year-old man and a 74-year-old man who had incidentally been indicated to have a right renal mass lesion by echography in other hospitals. In both cases, enhanced computed tomographic scan showed a low density renal mass. In selective renal angiogram of the two cases, a spoke-wheel configuration of vessels could be seen in one case, but there was no evidence of renal mass in another. Total nephrectomy was performed in each case. Two masses were pathologically diagnosed as oncocytoma, constructed of large eosinophilic cells with granular cytoplasm and small regular nuclei. These two patients have been well for more than 2 years after the surgery. These are the 25th and 26th cases of renal oncocytoma reported in Japan before December, 1988.

[Torsion of the undescended testis: report of two cases].

We report the torsion of the inguinal undescended testis seen in two boys 9 and 11 years old. Orchiopexy was performed in each case. We discussed the preoperative diagnosis and treatment of this disease. Orchiectomies are still frequently performed, because the preoperative diagnosis is often delayed.

[Fungal urinary infection: report no. 1].

Clinical studies were performed on 115 fungal strains isolated from the urine of 36 patients. The predisposing factors were fungal urinary tract infections, antibiotics and indwelling catheter. Forty five strains of the 115 strains were Candida albicans, the most prevalent, followed by 25 strains of C. tropicalis and 22 strains of Trichosporon beigelii. Species of Torulopsis were rare, only 5 strains were isolated. Sixty strains (56%) co-existed with bacteria in urine. The co-existing bacteria most frequently isolated were Pseudomonas and Streptococcus. The minimum inhibitory concentration (MIC) of 5-FU was measured using the Bact-Morphology Agar medium. C. albicans, C. guillermondii, and Torulopsis glabrata showed an excellent MIC (under 0.78 microgram/ml), C. tropicalis, C. parapsilosis, T. beigelii, and T. cutaneum showed a poor MIC (over 100 micrograms/ml).

[Initial clinical experience with video-cystoscope].

The endoscopic image processing system which has a very small change coupled device (CCD) at the distal tip of an endoscope, can give us a quite different imaging information from the conventional optical endoscopes, of which the main functions were just to "see inside the human body". The advantage of this endoscopic image processing system has been well recognized but, since the scope diameter could not be made smaller due to the size limitations of the CCD chip itself, the system has not been utilized in the field of Urology. In cooperation with Olympus Optical Co., we have studied a system called Urological Video Information System (UVIS) in order to utilize the image processing system for Urology. In our system a special light source with RGB light output is utilized together with a conventional optical cystoscope and a video converter is connected to the eyepiece of the scope in order to observe endoscopic images on the monitor. Endoscopic images can be stored in an image filing system when necessary. The image quality of UVIS is inferior to that of the conventional cystoscopes at the moment and there are several other technical problems to solve but, as witnessed in the field of Gastroenterology, it is expected that this kind of electronic system will become much more important in the future. This report covers the current problems and some considerations of them as an initial study.

[Application of video endoscope system (EVIS200) in endourology].

We began clinical studies in April, 1989 for the purpose of utilizing the video endoscope system in the field of urology, termed the Urological Video Information System (UVIS). The UVIS is made up of an image assembling and recording system and an image filling system. Since we started using the first generation video endoscope system (EVIS-1, Olympus Optical Co.) as an image assembling system, we have encountered several major obstacles such as 1) the absolute insufficiency for the amount of light, and 2) the unclearness of frozen images. As we have been able to use as second generation video endoscope system (EVIS200, Olympus Optical Co.) since September 1990, the major components of the UVIS have been modified. In this report, we describe our clinical experience with the re-designed UVIS; EVIS200, which we have used, is composed of a video-processor (CL-200, Olympus Co.), a video converter and rigid urological scopes. The EVIS200 has been evaluated not only for the endoscopic examination but also for transurethral surgery. No problems were noticed in manipulating the EVIS200. Marked improvements were achieved in image quality. Images both on the television monitor and hard copies from the color video printer had optical quality in good agreement with the gross findings. The UVIS was a great contribution to both the authors and the patients; the findings both of the examination and the surgery could be explained to the patient and family during and/or immediately after the procedure. We believe that the UVIS may replace conventional urologic endoscopy and become an excellent date-base for endourology.