Nobuoki Kohno

Prospective versus Retrospective ECG-gated 64-Detector Coronary CT Angiography: Assessment of Image Quality, Stenosis, and Radiation Dose

PURPOSE To show that prospective electrocardiographically (ECG)-triggered coronary computed tomographic (CT) angiography (hereafter, prospective CT angiography) is at least as effective as retrospective ECG-gated coronary CT angiography (hereafter, retrospective CT angiography). MATERIALS AND METHODS Institutional review committee approval and informed consent were obtained. Sixty patients with heart rates of less than 75 beats per minute who were referred for coronary CT angiography were enrolled. Both prospective and retrospective CT angiography were performed with a 64-detector scanner. Data acquisition times were recorded. Two independent cardiac radiologists evaluated subjective image quality (1, excellent; 4, poor) and severity of stenosis (0% occlusion, 1%-49% occlusion, 50%-75% occlusion, and >75% occlusion) with the 17-segment American Heart Association classification model. Discrepancies were settled by consensus. Effective radiation doses of prospective and retrospective CT angiography were calculated with volume CT dose index. Data regarding acquisition time and radiation exposure for prospective and retrospective CT angiography were compared. The Student t test was performed, and kappa statistics were calculated. RESULTS Mean data acquisition time of prospective CT angiography was shorter than that of retrospective CT angiography (5.6 seconds +/- 1.1 [standard deviation] vs 6.7 seconds +/- 1.1, respectively; P < .01). Consensus-determined image quality in coronary artery branches was similar between prospective CT angiography and retrospective CT angiography (1.15 vs 1.13, respectively; P = .992). Excellent agreement between prospective CT angiography and retrospective CT angiography was observed in the detection of significant (>or=50% occlusion) coronary artery stenoses per segment (kappa = 0.882) and in the grading of stenoses per patient (kappa = 0.829). Calculated effective dose with prospective CT angiography was 79% lower than that with retrospective CT angiography (4.1 mSv +/- 1.8 vs 20.0 mSv +/- 3.5, respectively; P < .001). CONCLUSION Prospective CT angiography can reduce radiation dose below that of retrospective CT angiography with dose modulation, while maintaining image quality and the ability to assess luminal obstructions in patients with heart rates of less than 75 beats per minute.

Increases of Amphiregulin and Transforming Growth Factor-{alpha} in Serum as Predictors of Poor Response to Gefitinib among Patients with Advanced Non-Small Cell Lung Cancers

Serum levels of amphiregulin and transforming growth factor-alpha (TGF-alpha), which were identified previously to be expressed at high levels in non-small cell lung cancer (NSCLC) with poor response to gefitinib, were examined by ELISA using blood samples taken from 50 patients with advanced NSCLCs. Of 14 cases that revealed above the cutoff line for amphiregulin in serum, 12 responded poorly to gefitinib, whereas 18 of the 36 cases showing below the cutoff revealed partial response (PR) or stable disease (SD; P = 0.026). Thirteen of 15 patients who were positive for TGF-alpha responded poorly to gefitinib, whereas 18 of the 35 patients with negative TGF-alpha levels turned out to be relatively good responders (P = 0.014). Of 22 patients with positive values for either or both markers, 19 were poor responders. On the other hand, among 28 patients negative for both markers, 17 were classified into the PR or SD groups (P = 0.001). Gefitinib-treated NSCLC patients whose serum amphiregulin or TGF-alpha was positive showed a poorer tumor-specific survival (P = 0.037 and 0.002, respectively, by univariate analysis) compared with those whose serum amphiregulin or TGF-alpha concentrations were negative. Multivariate analysis showed an independent association between positivity for TGF-alpha and shorter survival times among NSCLC patients treated with gefitinib (P = 0.034). Amphiregulin or TGF-alpha positivity in NSCLC tissues was significantly higher in male, nonadenocarcinomas, and smokers. Our data suggest that the status of amphiregulin and TGF-alpha in serum can be an important predictor of the resistance to gefitinib among patients with advanced NSCLC.

Pulse Wave Velocity Predicts Cardiovascular Mortality

Background Arterial stiffness measurements, generally from pulse wave velocity (PWV), are widely used with little knowledge of their relationship to long-term cardiovascular mortality in general populations. Methods and Results We studied a cohort of 492 Japanese-Americans living in Hawaii (mean age: 63.7 ±8.8 years) to assess the relationship between PWV and cardiovascular disease mortality and all-cause mortality. During the 10-year follow-up, 43 patients died (14 from cardiovascular events). The cohort was divided into 2 groups by the cut-off value of PWV (9.9 m/s) represented in the receiver operating characteristic curve. The risk ratio for PWV values >9.9 m/s to all-cause mortality was 1.28 [95% confidence interval (CI): 1.14-1.42], and adjusted for other risk factors this ratio was 1.42 (95% CI: 0.96-2.11). The corresponding risk ratios for cardiovascular mortality was 4.46 (95% CI: 1.61-12.32) and 4.24 (95% CI: 1.39-12.96), respectively. Conclusions The present study demonstrated that an increased PWV value is associated with future cardiovascular disease death in Japanese-Americans living in Hawaii. (Circ J 2005; 69: 259 - 264)

Characterization of Noncalcified Coronary Plaques and Identification of Culprit Lesions in Patients With Acute Coronary Syndrome by 64-Slice Computed Tomography

OBJECTIVES We sought to characterize noncalcified coronary atherosclerotic plaques in culprit and remote coronary atherosclerotic lesions in patients with acute coronary syndrome (ACS) with 64-slice computed tomography (CT). BACKGROUND Lower CT density, positive remodeling, and adjacent spotty coronary calcium are characteristic vessel changes in unstable coronary plaques. METHODS Of 147 consecutive patients who underwent contrast-enhanced 64-slice CT examination for coronary artery visualization, 101 (ACS; n = 21, non-ACS; n = 80) having 228 noncalcified coronary atherosclerotic plaques (NCPs) were studied. Each NCP detected within the vessel wall was evaluated by determining minimum CT density, vascular remodeling index (RI), and morphology of adjacent calcium deposits. RESULTS The CT visualized more NCPs in ACS patients (65 lesions, 3.1 +/- 1.2/patient) than in non-ACS patients (163 lesions, 2.0 +/- 1.1/patient). Minimum CT density (24 +/- 22 vs. 42 +/- 29 Hounsfield units [HU], p < 0.01), RI (1.14 +/- 0.18 vs. 1.08 +/- 0.19, p = 0.02), and frequency of adjacent spotty calcium of NCPs (60% vs. 38%, p < 0.01) were significantly different between ACS and non-ACS patients. Frequency of NCPs with minimum CT density <40 HU, RI >1.05, and adjacent spotty calcium was approximately 2-fold higher in the ACS group than in the non-ACS group (43% vs. 22%, p < 0.01). In the ACS group, only RI was significantly different between 21 culprit and 44 nonculprit lesions (1.26 +/- 0.16 vs. 1.09 +/- 0.17, p < 0.01), and a larger RI (> or = 1.23) was independently related to the culprit lesions (odds ratio: 12.3; 95% confidential interval: 2.9 to 68.7, p < 0.01), but there was a substantial overlap of the distribution of RI values in these 2 groups of lesions. CONCLUSIONS Sixty-four-slice CT angiography demonstrates a higher prevalence of NCPs with vulnerable characteristics in patients with ACS as compared with stable clinical presentation.

PPAR- γ agonist attenuates renal interstitial fibrosis and inflammation through reduction of TGF- β

Thiazolidinediones (TZDs), synthetic peroxisome proliferator-activated receptor (PPAR)-γ ligands, have a central role in insulin sensitization and adipogenesis. It has been reported that TZDs exert protective effects in both diabetic and nondiabetic models of renal disease, although the exact mechanism is not well understood. In particular, only a few studies have reported the renoprotective effects of TZDs in nondiabetic models of tubulointerstitial fibrosis and inflammation. Therefore, we investigated the anti-fibrotic and anti-inflammatory effects of the TZD troglitazone in the mouse model of unilateral ureteral obstruction (UUO). C57BL/6J mice underwent UUO and were studied after 3 and 7 days. Animals were divided into three groups and received control vehicle, troglitazone (150 mg/kg per day) or troglitazone (300 mg/kg per day) by gavage. Kidneys were harvested for morphological, mRNA and protein analysis. Reverse-transcriptase–PCR was used to assess the expression of transforming growth factor-β1 (TGF-β1) and the TGF-β1 type I receptor (TGFβR-I). Protein expression was assessed by western blotting (TGFβR-I) and immunostaining (TGFβR-I, α-smooth muscle actin (α-SMA), type I collagen (collagen I), F4/80, and proliferating cell nuclear antigen (PCNA)). The expression of α-SMA, collagen I, and F4/80 was decreased in mice treated with troglitazone compared with the control group. The numbers of PCNA-positive interstitial cells were decreased in mice treated with troglitazone. TGF-β1 mRNA and TGFβR-I mRNA and protein expression were decreased in the group treated with troglitazone compared with the control group. The beneficial effects of troglitazone treatment were also dose dependent. PPAR-γ agonist significantly reduced TGF-β and attenuated renal interstitial fibrosis and inflammation in the model of UUO.

Circulating visfatin level is correlated with inflammation, but not with insulin resistance

Objective  Recent studies, both in vitro and in vivo, have indicated that visfatin is one of the inflammatory cytokines, although the relationship between visfatin and insulin resistance remains inconclusive. Accordingly, we assessed the association between visfatin concentrations in serum and those of interleukin‐6 (IL‐6) and C‐reactive protein (CRP), known as markers of systemic inflammation, and also investigated the relationship between these serum concentrations and insulin resistance.

ADAM8 as a Novel Serological and Histochemical Marker for Lung Cancer

Purpose and Experimental Design: We have been investigating genes involved in pulmonary carcinogenesis by examining gene expression profiles of non–small-cell lung cancers to identify molecules that might serve as diagnostic markers or targets for development of new molecular therapies. A gene encoding ADAM8, a disintegrin and metalloproteinase domain-8, was selected as a candidate for such molecule. Tumor tissue microarray was applied to examine expression of ADAM8 protein in archival lung cancer samples from 363 patients. Serum ADAM8 levels of 105 lung cancer patients and 72 controls were also measured by ELISA. A role of ADAM8 in cellular motility was examined by Matrigel assays. Results: ADAM8 was abundantly expressed in the great majority of lung cancers examined. A high level of ADAM8 expression was significantly more common in advanced-stage IIIB/IV adenocarcinomas than in adenocarcinomas at stages I–IIIA. Serum levels of ADAM8 were significantly higher in lung cancer patients than in healthy controls. The proportion of the serum ADAM8-positive cases defined by our criteria was 63% and that for carcinoembryonic antigen was 57%, indicating equivalent diagnostic power of these two markers. A combined assay using both ADAM8 and carcinoembryonic antigen increased sensitivity because 80% of the lung cancer patients were then diagnosed as positive, whereas only 11% of 72 healthy volunteers were falsely diagnosed as positive. In addition, exogenous expression of ADAM8 increased the migratory activity of mammalian cells, an indication that ADAM8 may play a significant role in progression of lung cancer. Conclusions: Our data suggest that ADAM8 should be useful as a diagnostic marker and probably as a therapeutic target.

A-Kinase Anchoring Protein AKAP220 Binds to Glycogen Synthase Kinase-3β (GSK-3β) and Mediates Protein Kinase A-dependent Inhibition of GSK-3β

Glycogen synthase kinase-3 (GSK-3) is regulated by various extracellular ligands and phosphorylates many substrates, thereby regulating cellular functions. Using yeast two-hybrid screening, we found that GSK-3β binds to AKAP220, which is known to act as an A-kinase anchoring protein. GSK-3β formed a complex with AKAP220 in intact cells at the endogenous level. Cyclic AMP-dependent protein kinase (PKA) and type 1 protein phosphatase (PP1) were also detected in this complex, suggesting that AKAP220, GSK-3β, PKA, and PP1 form a quaternary complex. It has been reported that PKA phosphorylates GSK-3β, thereby decreasing its activity. When COS cells were treated with dibutyryl cyclic AMP to activate PKA, the activity of GSK-3β bound to AKAP220 decreased more markedly than the total GSK-3β activity. Calyculin A, a protein phosphatase inhibitor, also inhibited the activity of GSK-3β bound to AKAP220 more strongly than the total GSK-3β activity. These results suggest that PKA and PP1 regulate the activity of GSK-3β efficiently by forming a complex with AKAP220.

Wnt Inhibitor Dickkopf-1 as a Target for Passive Cancer Immunotherapy

Dickkopf-1 (DKK1) is an inhibitor of Wnt/beta-catenin signaling that is overexpressed in most lung and esophageal cancers. Here, we show its utility as a serum biomarker for a wide range of human cancers, and we offer evidence favoring the potential application of anti-DKK1 antibodies for cancer treatment. Using an original ELISA system, high levels of DKK1 protein were found in serologic samples from 906 patients with cancers of the pancreas, stomach, liver, bile duct, breast, and cervix, which also showed elevated expression levels of DKK1. Additionally, anti-DKK1 antibody inhibited the invasive activity and the growth of cancer cells in vitro and suppressed the growth of engrafted tumors in vivo. Tumor tissues treated with anti-DKK1 displayed significant fibrotic changes and a decrease in viable cancer cells without apparent toxicity in mice. Our findings suggest DKK1 as a serum biomarker for screening against a variety of cancers, and anti-DKK1 antibodies as potential theranostic tools for diagnosis and treatment of cancer.

DEC1 Modulates the Circadian Phase of Clock Gene Expression

ABSTRACT DEC1 suppresses CLOCK/BMAL1-enhanced promoter activity, but its role in the circadian system of mammals remains unclear. Here we examined the effect of Dec1 overexpression or deficiency on circadian gene expression triggered with 50% serum. Overexpression of Dec1 delayed the phase of clock genes such as Dec1, Dec2, Per1, and Dbp that contain E boxes in their regulatory regions, whereas it had little effect on the circadian phase of Per2 and Cry1 carrying CACGTT E′ boxes. In contrast, Dec1 deficiency advanced the phase of the E-box-containing clock genes but not that of the E′-box-containing clock genes. Accordingly, DEC1 showed strong binding and transrepression on the E box, but not on the E′ box, in chromatin immunoprecipitation, electrophoretic mobility shift, and luciferase reporter assays. Dec1−/− mice showed behavioral rhythms with slightly but significantly longer circadian periods under conditions of constant darkness and faster reentrainment to a 6-h phase-advanced shift of a light-dark cycle. Knockdown of Dec2 with small interfering RNA advanced the phase of Dec1 and Dbp expression, and double knockdown of Dec1 and Dec2 had much stronger effects on the expression of the E-box-containing clock genes. These findings suggest that DEC1, along with DEC2, plays a role in the finer regulation and robustness of the molecular clock.