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.

A new opto-mechanical effect in solids.

We propose that large, reversible shape changes in solids, of between 10%-400%, can be induced optically by photoisomerizing monodomain nematic elastomers. Empirical and molecular analysis of shape change and its relation to thermal effects is given along with a simple model of the dynamics of response. Our experiments demonstrate these effects for the first time and theory is compared qualitatively with our results.

[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.

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.

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.

Increase of calmodulin III gene expression by mu-opioid receptor stimulation in PC12 cells.

Calmodulin (CaM) is a principal multifunctional mediator of Ca2+ signaling in cells. It is reported that morphine increases CaM contents in mouse brain. However, the precise mechanism of CaM induction by morphine is unknown. We investigated the changes of CaM by opioid receptor stimulation in mRNA and protein levels. Expression of CaM was increased in dose- and time-dependent manners by morphine with RT-PCR assay in PC12 cells, and naloxone inhibited the effect of morphine. The expression was also increased with DAMGO (mu-opioid agonist), but not by DPDPE (delta) and U50488 (kappa). Northern blot analysis revealed that the CaMIII gene was responsive to morphine or DAMGO. CaM protein increased by DAMGO were distributed in both soluble and membranous fractions in the cells. Taken together, the data suggest that morphine induces the expression of CaMIII gene through mu-opioid receptor stimulation.

Complementary DNA Cloning and Characterization of Pearlin, a New Class of Matrix Protein in the Nacreous Layer of Oyster Pearls.

Calcified shell layer is composed of two polymorphs of CaCO(3), aragonite or calcite, and an organic matrix. The organic matrix consists of EDTA-soluble and insoluble fractions. These fractions are thought to regulate the formation of the elaborate shell structure. After decalcification of powdered pearl with 0.3 M EDTA, an EDTA-insoluble fraction was extracted with 0.3 M EDTA/8 M urea. This extraction step enabled us to purify a new class of EDTA-insoluble protein, Pearlin, almost homogeneously. Pearlin has a molecular weight of about 15 kDa and contains a sulfated mucopolysaccharide. We cloned the complementary DNA coding for Pearlin and deduced its complete amino acid sequence. Sequence analysis reveals that Pearlin is composed of 129 amino acids with a high proportion of Gly (10.8%), Tyr (10.0%), Cys (8.5%), Asn (7.7%), Asp (7.7%), and Arg (7.7%). Northern blot analysis showed that Pearlin messenger RNA was expressed specifically in mantle epithelium. From the sequencing data, Pearlin was shown to be quite different from the fibrous protein rich in Ala and Gly. The function of this protein in biomineralization is discussed.

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.

Calmodulin functions as an activator of Pur alpha binding to single-stranded purine-rich DNA elements (PUR elements).

Pur alpha is a single stranded DNA-binding protein and binds to a consensus sequence (GGN)n. We have reported that the DNA-binding activity of a single stranded cyclic AMP response element-binding protein (ssCRE-BP) is suppressed in cerebellum treated chronically with morphine, ssCRE-BP is identical to Pur alpha and the DNA binding activity of Pur alpha is markedly enhanced by a heat stable activator in the nuclear extract. In this report, we purified this activator. The amino acid composition and partial amino acid sequence were determined to be identical to those of calmodulin (CaM), which enhanced the binding of GST-Pur alpha to various PUR elements in the 5' non-coding regions of the neuropeptide Y, myelin basic protein and nicotinic Ach receptor beta 4 subunit genes. The data suggest a novel gene expression pathway mediated by Ca/CaM-Pur alpha which may regulate a variety of genes in addition to those regulated through the CREB pathway.

Relationship between calcium channel antagonists and nocturnal hypotension and autonomic imbalance in patients with a previous myocardial infarction.

The present study was conducted to evaluate the effect of calcium channel antagonists on diurnal changes in blood pressure and on autonomic function in 71 patients who were receiving a calcium channel antagonist because they had previously suffered a myocardial infarction. Ambulatory blood pressures and Holter ECGs were recorded simultaneously for 24 h. Autonomic function was assessed by heart rate variability. Nocturnal systolic pressure was > or = 90 mmHg in 63 patients (group I) and < 90 mmHg in 8 patients (group II). Significant day to night changes in high-frequency power (from 4.3 +/- 1.2 to 4.5 +/- 1.0/m2) as well as in the ratio of low-frequency power to high-frequency power (from 1.3 +/- 0.1 to 1.1 +/- 0.2) were observed in group I. whereas such changes were blunted in group II. When the calcium antagonist was discontinued or the dose was reduced in group II, the autonomic imbalance improved along with elevation of nocturnal systolic blood pressure. Thus, nocturnal blood pressure should be monitored when such drugs are administered for the treatment of ischemic heart disease to a patient with a previous myocardial infarction. If nocturnal hypotension occurs, the dose should be reduced or the drug should be discontinued.

Hormonal and cardiorenal responses to acute saline loading in mild congestive heart failure--the effect of angiotensin converting enzyme inhibition.

We investigated cardiorenal responses to acute saline loading in patients with mild congestive heart failure (CHF) with and without angiotensin-converting enzyme (ACE) inhibition. Saline loading (infusion of 250 ml of normal saline) was performed on 10 patients with mild CHF and 10 control subjects. Although saline loading reduced plasma renin activity and plasma angiotensin II to a similar extent in both groups, it increased cardiac output, renal blood flow, and the ratio of renal blood flow to cardiac output in the mild CHF group but not in the control group. After saline loading, urinary sodium excretion was higher in the control than in the mild CHF group. In the mild CHF group, saline loading was performed again after an ACE inhibitor (delapril) had been administered for 5 days. Although delapril increased cardiac output and renal blood flow under basal conditions, saline loading did not affect these variables. Delapril improved urinary sodium excretion after saline loading. Thus, the renin-angiotensin-aldosterone axis may contribute to cardiorenal hemodynamics and renal sodium handling in patients with mild CHF. This effect may attenuate the natriuretic response to acute saline loading, and administration of an ACE inhibitor improves this attenuation.

[Studies on residual cardiovascular dysfunction in patients receiving long-term antihypertensive treatment of calcium channel blockers: with special reference to left ventricular hypertrophy, and impairments of left ventricular diastolic function and carotid arterial distensibility].

Residual cardiovascular dysfunctions including left ventricular hypertrophy, and impairment of left ventricular diastolic function and carotid arterial distensibility were investigated in hypertensive patients treated with calcium channel blockers for more than 1 year. Ultrasonographic examinations of the heart and carotid artery were performed in patients treated with calcium channel blocker alone for more than 1 year (n = 45) and in age-, sex- and weight-matched control subjects (n = 29). The following parameters were obtained: left ventricular mass index, cardiac diastolic function (A/E ratio) and carotid arterial distensibility (Distens). Hypertensive subjects were re-examined under the same conditions with the same parameters after 10 +/- 5 months. Patients with hypertension revealed no significant changes in these three parameters after 10 +/- 5 months Patients with left ventricular hypertrophy (n = 20) revealed significant impairments in diastolic function and carotid arterial distensibility (A/E = 1.42 +/- 0.25, Distens = 2.4 +/- 1.3% kPa) compared to those without left ventricular hypertrophy (n = 25) (A/E = 1.18 +/- 0.29, Distens = 3.8 +/- 1.7% kPa, p < 0.05). Patients without left ventricular hypertrophy had significantly impaired cardiovascular functions compared to the normal control group (A/E = 1.03 +/- 0.27, Distens = 6.3 +/- 2.2% kPa, p < 0.05, p < 0.01 respectively). Therefore, only reduction of blood pressure with calcium channel blocker may not be enough to improve cardiovascular organ damage, especially in patients with residual left ventricular hypertrophy, and such residual functional deteriorations must be corrected probably with another pharmaceutical modality.

Safety and effectiveness of exercise training in patients with silent myocardial ischemia.

The effectiveness of exercise training in patients with silent myocardial ischemia was examined. Forty patients with coronary heart disease (mean age 55 +/- 8 years) were recruited for a 12-week exercise training program. All patients underwent treadmill exercise stress testing, exercise thallium-201 single photon emission computed tomography and left heart catheterization. They were divided into three groups based on the symptoms and the results of exercise thallium scintigraphy, i.e., painful myocardial ischemia (PMI group), silent myocardial ischemia (SMI group), and non-myocardial ischemia (NMI group). Normalized treadmill time was longer in the SMI group (108 +/- 24%) than in the PMI group (86 +/- 14%, p < 0.05). All 40 patients, 14 from the PMI group, 16 from the SMI group and 10 from the NMI group, completed the whole exercise training program. A significant prolongation of treadmill time was attained in all three groups after exercise training [PMI group: from 494 +/- 105 to 632 +/- 78 sec (p < 0.05), SMI group: from 609 +/- 147 to 746 +/- 137 sec (p < 0.05), NMI group: from 572 +/- 112 to 739 +/- 13 sec (p < 0.05)]. The improvement of myocardial ischemia following exercise training was similar in the SMI and PMI groups. No adverse effects were detected throughout the program. The exercise training program adopted in this study proved safe and effective in patients with silent myocardial ischemia.

Serum prostate-specific antigen values for the prediction of clinical stage and prognosis in patients with prostate cancer: an analysis of 749 cases.

BACKGROUND: The clinical significance of pretreatment serum prostate-specific antigen (PSA) values was studied to determine the ability to predict clinical stage and prognosis using a relatively large number of patients with prostate cancer. METHODS: Serum PSA values at diagnosis were analyzed from 749 patients with newly-diagnosed prostate cancer and registered in the Tokai Urological Cancer Registry. Correlations between the PSA value, the clinical stage and prognosis of the patients were evaluated. RESULTS: Serum PSA values at each stage of diagnosis showed positivity (> or = 3.6 ng/mL) in 23% (stage A1) to 91.2% (stage D2) of patients, and it was possible to obtain statistical differences between the stages, even between A1 and A2. Based on a cumulative study of PSA distribution, stages greater than A2 could be diagnosed using a cut-off of 7.2 ng/mL, with a 99.2% positive predictive value (PPV), and a 16.2% negative predictive value (NPV). At a PSA level of 10.8 ng/mL, stages greater than B2 could be predicted with a PPV of 95.3% but an NPV of 40.3%. Pretreatment PSA values were a significant prognostic indicator in stage D2 patients using 100 to 150 ng/mL as the cut-off values. These differences were primarily found in the poorly differentiated group, which showed a statistical difference using cut-off PSA values from 75 to 150 ng/mL. CONCLUSIONS: Serum PSA levels from a large number of patients can be used to predict the stage and prognosis of prostate cancer patients.

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.

Focally spared area of fatty liver caused by arterioportal shunt.

We describe a case with a focally spared area in fatty liver caused by arterioportal shunt. Furthermore, we discuss the cause of the focally spared area related to a localized dilution or reduction in portal blood flow.

[Evaluation of the blood flow in tumors before and after irradiation with color Doppler flow imaging].

The blood flow in tumors of three patients with lung cancer, cancer in oral cavity and metastatic lymphnode was respectively evaluated before and after irradiation with color Doppler flow imaging. The arterial pulsating flow in lung cancer disappeared after 30 Gy irradiation and that in cancer of oral cavity disappeared after 15 Gy irradiation with intraarterial infusion chemotherapy. But the arterial pulsating flow in metastatic lymphnode was detected during irradiation and disappeared after 40 days post- irradiation. Therefore color Doppler flow imaging seemed to be useful for evaluation of the blood flow in tumors before and after irradiation.

[Clinical experience with extracorporeal shock wave lithotripsy MFL-5000 for urinary stone].

A Dornier MFL 5000, a new generation extracorporeal shock wave lithotripter, was installed in our hospital and the first Japanese clinical experience has been collected between July and November in 1989. We report our experience with the first 35 patients with 45 stones who were treated in 42 treatments using ESWL. We followed up 3 weeks. No invasive anesthesia was performed except 2 cases of epidural anesthesia. A double J catheter was installed in 2 patients, a ureteral catheter in 4 patients, and PNL was performed in 2 patients before ESWL. We did not use a PNL or a TUL in the postoperative treatment. In the 3 week followed up period, 29 patients (82.9%) were completely free from stone fragments. No serious complications were observed after ESWL. We conclude that the DORNIER MFL 5000 is effective for renal and ureteral stones without serious complications.