Apoptosis induction by antisense oligonucleotides against miR-17-5p and miR-20a in lung cancers overexpressing miR-17-92.

Amplification and overexpression of the miR-17-92 microRNAs (miRNA) cluster at 13q31.3 has recently reported, with pointers to functional involvement in the development of B-cell lymphomas and lung cancers. In the present study, we show that inhibition of miR-17-5p and miR-20a with antisense oligonucleotides (ONs) can induce apoptosis selectively in lung cancer cells overexpressing miR-17-92, suggesting the possibility of 'OncomiR addiction' to expression of these miRNAs in a subset of lung cancers. In marked contrast, antisense ONs against miR-18a and miR-19a did not exhibit such inhibitory effects, whereas inhibition of miR-92-1 resulted in only modest reduction of cell growth, showing significant distinctions among miRNAs of the miR-17-92 cluster in terms of their roles in cancer cell growth. During the course of this study, we also found that enforced expression of a genomic region, termed C2, residing 3' to miR-17-92 in the intron 3 of C13orf25 led to marked growth inhibition in association with double stranded RNA-dependent protein kinase activation. Finally, this study also revealed that the vast majority of C13orf25 transcripts are detected as Drosha-processed cleavage products on Northern blot analysis and that a novel polyadenylation site is present 3' to the miR-17-92 cluster and 5' to the C2 region. Taken together, the present findings contribute towards better understanding of the oncogenic roles of miR-17-92, which might ultimately lead to the future translation into clinical applications.

Carotid arterial distensibility is an important determinant of improvement in autonomic balance after successful coronary angioplasty.

OBJECTIVE: To determine whether arterial distensibility influences improvement of autonomic imbalance after left ventricular wall motion (LVWM) abnormality has been improved by percutaneous transluminal angioplasty (PTCA) in patients with ischemic heart disease (IHD). METHODS AND RESULTS: The following variables were assessed before and 3 months after PTCA in 63 patients with IHD who had a successful PTCA: heart rate variability (standard deviation of all normal R-R intervals [SDNN] measured from 24 h Holter electrocardiograms), baroreceptor sensitivity (determined by the phenylephrine injection method), and carotid artery distensibility (determined by ultrasonography). Abnormalities of LVWM were assessed by cineventriculography. In a multivariate analysis, carotid artery distensibility before PTCA and changes in LVWM abnormality before and after PTCA were independently associated with changes in baroreceptor sensitivity and SDNN (P< 0.05). Patients were divided into two groups: one with impaired and one group with unimpaired initial carotid artery distensibility. After PTCA, LVWM was improved significantly in both groups, but baroreceptor sensitivity and SDNN were improved significantly (respectively from 5.0 +/- 3.3 to 5.9 +/- 3.9 ms/mmHg, P< 0.01 and from 111 +/- 47 to 128 +/- 54 ms, P < 0.01) only in patients with well-preserved carotid artery distensibility. CONCLUSIONS: The beneficial effect of PTCA on autonomic balance was greater in patients with well-preserved carotid artery distensibility than in those with impaired carotid artery distensibility. These results suggest that the pathophysiological state of arterial distensibility may modify the autonomic balance even after myocardial ischemia-related LVWM abnormalities are improved.

An impaired carotid sinus distensibility and baroreceptor sensitivity alter autonomic activity in patients with effort angina associated with significant coronary artery disease.

Baroreceptor sensitivity and carotid sinus distensibility were lower in patients with angina associated with significant coronary artery disease than in patients with vasospastic angina. Baroreceptor sensitivity was significantly correlated with carotid sinus distensibility in both groups of patients.

Puralpha, a single-stranded DNA binding protein, suppresses the enhancer activity of cAMP response element (CRE).

Puralpha, a single-stranded DNA binding protein, recognizes a PUR element (GGN repeat). We have reported that Puralpha binds to a single-stranded oligonucleotide probe containing the cAMP response element (CRE) of rat somatostatin gene using a gel mobility shift assay. Here, we showed that Puralpha binds to the probe only in the presence of a PUR element by a more detailed characterization. We also examined the effects of Puralpha on the enhancer activity of the somatostatin CRE in PC12 cells using the reporter gene assay. Transfected Puralpha suppressed the CRE enhancer activity stimulated by forskolin (which increases intracellular cAMP), but suppression was not observed when the PUR element was deleted. The neurite extension induced by forskolin was inhibited by the transfection of Puralpha, but that by NGF was not suppressed. The c-fos mRNA induced by forskolin, but not by NGF, was also suppressed by Puralpha transfection. These results indicate that Puralpha suppresses the biological activities induced by forskolin, but not by NGF, in PC12 cells and that Puralpha could interfere with a cAMP-CRE signal pathway.

Characteristics of cardiovascular morphology and function in the high-normal subset of hypertension defined by JNC-VI recommendations.

A cross-sectional study was conducted to compare the morphological and functional characteristics of the cardiovascular system among subgroups of hypertension defined by the JNC-VI recommendations. One hundred and sixteen subjects (normotensives and unmedicated hypertensives: 49+/-10 yr) were classified into 4 groups based on the criteria of JNC-VI: normotensive (NOR: n = 38), high-normal blood pressure (HN: n = 16), stage 1 hypertensive (SI: n = 28), and stage 2 to 3 hypertensive (SII-III: n = 34). Ultrasonographic examinations of the heart and carotid artery were performed in all subjects, and the following parameters were obtained: left ventricular mass index (LVMI), relative wall thickness at end-diastole (RWTd), cardiac diastolic function (A/E), common carotid artery diameter (CAD), intimal media thickness of the common carotid artery (IMT), and distensibility of the common carotid artery (Distens). RWTd, A/E, and IMT in SI (RWTd, 0.41+/-0.07; A/E, 1.21+/-0.41; IMT, 0.69+/-0.17 mm) and SII-III patients (0.40+/-0.08, 1.38+/-0.33, 0.80+/-0.21 mm) were larger than those in NOR patients (0.33+/-0.03, 0.86+/-0.21, 0.56+/-0.10 mm) (p < .01). Furthermore, LVMI in SII-III (135.5+/-35.5 g/m2) patients was larger than that in NOR patients (99.4+/-17.5 g/m2) (p < .05). RWTd in HN patients (0.37+/-0.06) was significantly higher than that in NOR patients (p < .05). A/E tended to be larger in HN than in NOR patients (p < 0.1). In the normotensives, no significant difference in any of the parameters was detected between those with optimal (n = 19) and normal (n = 19) blood pressure. Thus, both morphological and functional changes were associated with elevation of blood pressure. Cardiac morphological adaptation and functional impairment were present even in subjects with high-normal blood pressure level, while there were no significant differences between the normal and optimal subsets.

[Clinical efficacy of fosfomycin in the treatment of orofacial infections].

In this study, the clinical efficacy of fosfomycin (FOM) for various types of orofacial infections was evaluated in 40 patients, 27 males and 13 females, ranging in age from 16 to 71 years. All but 1 patients were treated orally with FOM at 500 mg 4 times daily. The total duration of treatment varied from 3 to 31 days. Basically, clinical effectiveness was determined by the criteria of Japanese Society of Oral and Maxillofacial Surgeons. Two cases, in which the final therapeutic evaluation could not be obtained because of side effects, were excluded from the analysis. The results were rated as excellent in 7, good in 26 and poor in 5 out of 38 evaluable cases, with an efficacy rate reaching 86.8%. From the clinical standpoint, the differences of efficacy were further discussed in terms of phase, type and intensity of infections respectively. In 9 (10 times) of 40 cases, the bacteriological analyses could be made. The identification of organisms obtained from abscess in each case demonstrated mixed infections caused by both aerobic and anaerobic organisms in more than half cases. From their sensitivity tests, it should be pointed out that FOM was efficiently active against aerobic and anaerobic Gram-positive organisms, while it was ineffective for the anaerobic Gram-negative organism, Bacteroides, although the role of the latter generally remained still obscure in orofacial infection. No serious side effects were observed except for mild diarrhea in 3 patients and a slight elevation of serum glutamic oxaloacetic transaminase (GOT) level in 1 patient. This study conclusively demonstrated that FOM was a useful and safe antibiotic in the treatment of patients with various orofacial infections.

[A case of transitional cell carcinoma of the bladder in a juvenile patient].

A 15-year-old male was referred to our hospital with the chief complaint of gross hematuria. Cystoscopic examination revealed a papillary tumor on the posterior wall. Transurethral resection of the bladder tumor was performed. Histological examination of the excised tumor showed transitional cell carcinoma, grade 1, pTa. Recurrence has not been observed for about 2 years after the operation. We investigated 54 previously reported Japanese cases of bladder cancer before age twenty including the present case.

[Alterations in urinary N-Acetyl-beta-D-glucosaminidase (NAG), beta 2-microglobulin (BMG) under acute exposure to simulated 6,000 m altitude].

The changes in NAG and BMG concentration in the urine during sudden exposure to a simulated 6,000 m altitude (354 mmHg) were studied. Subjects were 5 healthy male volunteers before a mountain climbing expedition (20-25 years old). Decompression was commenced at 11:00 and terminated at 17:00 (2-h ascent, 2-h sojourn, 2-h return) for 5 successive days. Urine was discarded at 9:00 and thereafter collected at 11:00 (group I), 14:00 (group II) 17:00 (group III), and 9:00 the next day (group IV) and urinary NAG and BMG was measured. The rate of NAG indicated elevation above 5.0 U/L in 2 cases in group III and elevation which was significantly different from that of the other 3 groups. In the other 3 cases, significant elevation was not observed, but for group III as a whole NAG excretion was significantly elevated compared of the other 3 groups. However, increased excretion of NAG was not observed on all 5 successive days. The change in BMG values was smaller than that of the NAG values and no significant elevation was observed in any of the 4 groups. Increasing urinary excretion of NAG by sudden exposure to a high altitude indicated excessive destroy of epithelium in the proximal tubules, but this change was reversible.

[Long ureteral polyp: a case report].

A case of long ureteral polyp is presented. The patient was a 62-year-old woman complaining of asymptomatic macrohematuria. Radiological examinations revealed ureteral tumor. A tumor was found with cystoendoscopy, and by transurethral biopsy in bladder the tumor was not malignant. Polypectomy was performed. The tumor removed was fibrous polyp measuring about 8 cm in length.

[Clinical evaluation of lymphoscintigraphy in urological disease].

Clinical efficacy of lymphoscintigraphy was evaluated in 31 cases of urological disease. To diagnose metastatic change of malignant tumor, 99mTc-rhenium colloid (5mCi) was administrated from bilateral pedal region. Of these patients 12 had true positive finding, 2 had false positive finding, 2 had false negative finding, and 8 had true negative finding defined by lymphoscintigram and pathological finding. Overall accuracy of lymphoscintigraphy revealed 83% in correlation of pathological finding, 78% in lymphangiography. Accuracy between lymphangiography and pathological finding was 80%. The overall clinical efficacy of lymphoscintigraphy was the same as lymphangiography. The merit of this method was that we can examine easily, painlessly, and frequently.

[A study on the levels of cefoperazone in urological organs].

The concentration of Cefoperazone (CPZ) in urological organs and serum was measured after the intravenous administration of 2 g of CPZ. Serum levels on healthy adults attained a maximal value of 122.1 +/- 4.2 micrograms/ml at 30 minutes. Serum levels of patients operated under general anesthesia attained a maximal value of 159.0 +/- 13.9 micrograms/ml at 30 minutes. Serum concentration on operated cases continued to be 1.3-2.2 fold higher than that of the healthy adults. Prostate tissue levels attained a maximal value of 80.9 +/- 3.6 micrograms/g at 30 minutes, vesical tissue levels were 52.1 +/- 1.3 microgram/g at 60 minutes and renal tissue were 94.0 +/- 14.5 micrograms/g at 90 minutes. Judging from the minimal inhibitory concentration of CPZ, CPZ seemed to be clinically useful in the treatment of urological infections.

[Detection of beta-lactamase-producing strains isolated from urinary tract and their drug susceptibility].

A total of 518 bacterial strains isolated from the urine of patients with various urological diseases in our Urological Department between November, 1987 and February, 1989 were studied for their beta-lactamase production and susceptibility to various antimicrobial agents was determined. beta-lactamase activity was determined by the acidometry disc method. There were 241 gram-positive cocci, 276 of gram-negative rods and 1 Neisseria gonorrhoeae. Thirty-four percent of the gram positive and 76.3% of gram negative rods produced beta-lactamase. S. aureus (81.3%), S. epidermidis (65.1%) in gram-positive cocci, E. cloacae (100%), S. marcescens (100%), C. freundii (100%), P. aeruginosa (97.2%), P. Rettgeri (88.9%), E. gergoviae (85.7%), K. oxytoca (84.6%), M. morganii (81.8%) and E. coli (69.0%) in gram-negative rods produced beta-lactamase at a higher rate. beta-lactamase produced by gram-positive cocci was entirely penicillinase, and that produced by gram-negative rods only penicillinase in 4.0%, only cephalosporinase in 44.2% and both in 25.4%. In S. aureus and S. epidermidis, the isolated rate of strains resistant to ampicillin (p less than 0.01) and piperacillin (p less than 0.05) in the beta-lactamase producing strains was significantly higher than that in the beta-lactamase non-producing strains. In E. coli, the isolation rate of strains resistant to ampicillin and piperacillin in the penicillinase-producing strains was significantly higher than in the penicillinase non-producing strains (p less than 0.01). But both cephalosporinase-producing strains and beta-lactamase non-producing strains showed high susceptibility to cephalothin. These results suggest that the penicillinase might present a clinical problem in the treatment of urinary tract infections by S. aureus, S. epidermidis and E. coli.

[The study of the ABH isoantigen of bladder tumors].

ABH isoantigens in 82 cases with bladder tumor of various grades and stages were investigated by the avidin-biotin-peroxidase complex (ABC) method. Correlation of ABC expression for isoantigens with pathological findings on the grading and staging system of the bladder tumors were as follow: Of 60 cases of low grade tumors, 34 (56.7%) were positive and of 22 cases of high grade tumors, 21 (95.5%) were negative. Of 63 cases of low stage tumors, 35 (55.6%) were positive and all 19 cases of high stage tumors, (100%) were negative. The isoantigens in the tissue of the bladder tumors tended to disappear as the histologic anaplasia of the tumor progressed. There was a high correlation between the histological grade, stage and ABC expression (p less than 0.01). In a follow-up study of 38 patients with low grade, low stage, initial transitional cell carcinoma, 9 of the 24 patients positive for ABC expression (37.5%) showed recurrence during the 1 to 6 year follow-up period, whereas 10 of the 14 patients negative for ABC expression 10 (71.4%) did. There was a high correlation between the recurrence rate and ABC expression (p less than 0.05). Therefore, the analysis in the bladder tumors may be valuable for prediction of malignant potential, especially for the low grade, low stage tumors.

[Study on heterotransplantation of human malignant urogenital tumors in nude mice: establishment of the tumor line in nude mice from human primary ureteral tumor and its characteristics].

The tumor line in nude mice is one of the most important experimental animal models for oncodevelopmental studies. We implanted a human ureteral tumor into nude mice, established the tumor line, AM-UT-1, and maintained it by serial transplantation. The characteristics of this tumor line in nude mice are reported. Histologically, the original tumor was a transitional cell carcinoma (grade II, stage B). The transplanted tumor grew locally and had a constant growth pattern during serial passage until the 13th passage in nude recipients and maintained the basic histological, immunohistochemical findings of the original tumor. Both original tumor and serial transplanted tumor had carcinoembryonic antigen (CEA)-producing activity. A high level of serum CEA was also recognized in nude mice.

The study on cell surface antigens in epithelial tumor of the upper urinary tract: ABH-isoantigen and Thomsen-Friedenreich antigen.

ABH-isoantigen (ABH-Ag) and Thomsen-Friedenreich antigen (T-Ag) were investigated by the Avidin-Biotin-Peroxidase Complex (ABC) method on 47 patients with epithelial tumor of the upper urinary tract (all patients underwent nephroureterectomy including the cuff of the bladder; 30 patients were diagnosed as transitional cell carcinoma of renal pelvis and 17 ureteral organs). The correlations between ABC expression for ABH-Ag and T-Ag with histological grade, stage and prognosis (5 year survival rate) were studied. A correlation was observed between grade (p less than 0.05) and deletion of the antigenicity of ABH-Ag, but no correlation was evident with stage and prognosis. A high correlation was evident, however, between grade (p less than 0.01), stage (p less than 0.01) and prognosis (p less than 0.01) and deletion of the antigenicity of T-Ag. The analysis of ABC expression for ABH-Ag and T-Ag may therefore be valuable for predicting the malignant potential in transitional cell carcinoma of the upper urinary tract. T-Ag determination in particular may provide a useful prognostic probe should it find clinical application.

[Chronological changes in bacteria isolated from the urinary tract and their drug sensitivities].

Statistical studies were performed on bacterial strains isolated from the urine of patients with various urological disease in our Urological Department between January, 1983 and December, 1987. In 1987, sensitivity tests of antibiotics to various pathogens were carried out. From the outpatients, 5,725 bacterial strains were isolated. S. epidermidis was isolated most frequently (23.8%), followed by Enterococcus species (16.5%), E. coli (14.7%), Streptococcus species (12.6%), Proteus species (5.3%), other GNR (4.6%), P, aeruginosa (4.5%), Klebsiella species (4.4%), and others. From the inpatients, 4,747 bacterial strains were isolated. Enterococcus species was isolated most frequently (23.1%), followed by S. epidermidis (18.7%), Fungi (12.9%), P. aeruginosa (8.0%), other GNR (6.8%), Streptococcus species (5.4%), Enterobacter species (4.8%), E. coli (4.1%), S. aureus (4.1%) and others. Annual change of distribution of organisms indicated that S. aureus from outpatients had a tendency to increase and Streptococcus species had a tendency to decrease. Whereas from inpatients, E. coli, other GNR, Pseudomonas species and Enterobacter species had a tendency to increase, and Streptococcus species, P. aeruginosa, Proteus species and Klebsiella species had a tendency to decrease. S. epidermidis showed a high sensitivity to minocycline and cephalothin, and Enterococcus showed a high sensitivity to ampicillin and minocycline, but did not show good sensitivity to cefoperazone and latamoxef. E. coli showed a high sensitivity to all drug we tested. P. aeruginosa showed a high sensitivity to gentamycin and norflaxacin, but did not show good sensitivity to other many drugs we tested.

[Study of the Thomsen-Friedenreich antigen in bladder tumors].

Thomsen Friedenreich antigen (T-ag), ABH isoantigen (ABH-ag) in 106 cases with bladder tumor (all transitional cell carcinoma) of various histological grades and stages were investigated by the Avidin-Biotin-Peroxidase Complex (ABC) method. There was a correlation between histological grade, stage and deletion of the antigenicity (T-ag and ABH-ag). In a follow-up study of 45 patients with low grade and low stage tumor, the recurrence rate after surgery of the cases with abnormal antigenicity (T-ag and ABH-ag) was significantly higher than that of the cases with the normal antigenicity. Combination of two markers (T-ag and ABH-ag) was significantly more effective than the single marker.

[Malignant mesodermal mixed tumor of the bladder: report of a case].

An 81-year-old woman was admitted to our clinic due to gross hematuria. A large bulky pedunculated mass was found in the bladder by cystoscopic examination. Subtotal cystectomy and bilateral cutaneostomy was performed on January 12, 1987. Histologically the tumor was composed of carcinomatous and sarcomatous elements. The carcinomatous element was composed fundamentally of grade 2, transitional cell carcinoma with numerous foci of squamous metaplasia. The sarcomatous element was composed of myxosarcomatous, chondro-sarcomatous pattern and non-differentiated malignant spindle cell component. Immunohistochemical examination demonstrated the presence of cytokeratin and epithelial membrane antigen in the spindle cell and more obvious carcinomatous regions, using the avidin-biotin conjugated immunoperoxidase technique. The patient died 3 months after operation. Autopsy findings showed multiple organ metastasis which were composed of carcinomatous and sarcomatous elements.

[clinical studies on primary renal pelvic tumors].

The 15 cases of the primary renal pelvic tumors treated at our Hospital between 1974 and 1983, were reviewed retrospectively. The patients ranged in age from 41 to 74 years old (average: 58.3 years old). There were 11 males and 4 females, the ratio being 2.8:1.0. The affected side was left in 9 cases and right in 6 cases. The most frequent symptom was macrohematuria, which was seen in 12 cases (80%). The major finding of IVP was non-functioning kidney, which was seen in 8 cases (53.3%). Positive urinary cytology was obtained in 8 cases (53.3%). As the surgical method, total nephroureterectomy with bladder cuff was performed in 8 cases, nephroureterectomy in one case and nephrectomy in 6 cases. Histologically, 14 cases were transitional cell carcinoma and one case was squamous cell carcinoma. Simultaneous urothelial tumors were seen in the bladder of 2 patients. A subsequent ureteral tumor was found in one of the 7 cases in which ureters were resected incompletely, and subsequent bladder tumors were found in 8 of the 15 cases receiving surgical treatment in the follow-up period. All of tumors were found within 2 years after operation. Over-all actual survival rates at 1, 3 and 5 years were 87%, 67%, 48%, respectively. Three and 5 year actual survival rates were 100%, 100% respectively for the low stage group and 59%, 29% respectively for the high stage group. Three and 5 year actual survival rates were 100%, 78%, respectively for the low grade group and 44%, 27% respectively for the high grade group. Among several factors, stage and grade of the tumor were the most influencing factors for prognosis.

[Clinical studies on primary ureteral tumors].

The 15 cases of the primary ureteral tumors treated at our Hospital between 1974 and 1983, were reviewed retrospectively. The incidence of primary ureteral tumors among the outpatients in our urologic clinic was 0.15%. The patients ranged in age from 50 to 75 years old (average: 65.5 years old). There were 11 males and 4 females, the ratio being 2.8:1.0. The right ureter and the lower third of the ureter were involved more frequently than other areas. The most frequent symptom was macrohematuria which was seen in 12 cases (80%). The major finding of IVP was non-functioning kidney, which was seen in 11 cases (73.3). Positive urinary cytology was obtained in 9 cases (60%). Twelve patients underwent nephroureterectomy with bladder cuff or total cystectomy. Histologically, all cases were transitional cell carcinoma. Simultaneous urothelial tumors were seen in the bladder in 4 cases (26.7%). The over-all actual survival rates at 1, 3 and 5 years were 59%, 42%, 42%, respectively. The 5-year actual survival rate was 63% for the low grade group and 0% for the high grade group. The 5-year actual survival rate was 82% for the low stage group and 0% for the high stage group. Among several factors, grade and stage of the tumor were the most influencing factors for prognosis.