Optimal strength and number of shocks at upper limit of vulnerability testing required to predict high defibrillation threshold without inducing ventricular fibrillation.

BACKGROUND: The upper limit of vulnerability (ULV) closely correlates with the defibrillation threshold (DFT). The aim of this study was to establish the optimal protocol for using the ULV test to predict high DFT (>20 J) without inducing ventricular fibrillation (VF). METHODS AND RESULTS: The 10-J and 15-J ULV test with 3 coupling intervals (-20, 0, and +20 ms to the peak of T-wave) and the DFT test were performed in 96 patients receiving implantable cardioverter defibrillator. ULV ≤ 10 J was confirmed in 47 (49%). ULV ≤ 15 J was confirmed in 70 (77%) of 91 patients (15-J ULV test could not be done in 5). The sensitivity and negative predictive value of both ULV >10 J and >15 J for predicting high DFT were 100%. The specificity and positive predictive value of ULV >15 J were higher than those for ULV >10 J (85% vs. 55%, 43% vs. 22%, respectively). The rate of VF inducibility for confirming ULV ≤ 15 J was lower than that for ULV ≤ 10 J (23% vs. 51%, P<0.0001). On analysis of single 15-J ULV test only at the peak of T-wave, VF was not induced in 79 of 91 patients, but 4 of these had high DFT. CONCLUSIONS: The 15-J ULV test with 3 coupling intervals could correctly identify high-DFT patients and reduce the necessity for VF induction at defibrillator implantation.

Rate-dependent and site-specific conduction block at the posterior right atrium and drug effects evaluated using a noncontact mapping system in patients with typical atrial flutter.

INTRODUCTION: Conduction block in the posterior right atrium (RA) plays an important role in perpetuating atrial flutter (AFL). Although conduction blocks have functional properties, it is not clear how the block line changes with the pacing rate, pacing site, and administration of antiarrhythmic drugs. METHODS AND RESULTS: Forty patients with typical AFL were enrolled. Pacing (110, 170, 230 ppm) from the coronary sinus ostium (CSo) and low lateral RA was performed. After 1 mg/kg pilsicainide (pure sodium channel blockade) administration, the pacing protocol was repeated. Conduction block was assessed based on a color-coded isopotential map and 20 points of virtual unipolar electrograms in the posterior RA using noncontact mapping. Block line proportion was defined as the percentage of length of the block line between the superior and inferior vena cava. The pacing rate-dependent extension of the block proportion was significant during pacing from both sides (pacing from the CSo: 59 ± 17% at 110 ppm, 69 ± 16% at 230 ppm, P < 0.05; pacing from the low lateral RA: 43 ± 19% at 110 ppm, 55 ± 22% at 230 ppm, P < 0.05). The block line was significantly longer during CSo pacing than during low lateral RA pacing at each rate (all P < 0.05). After pilsicainide administration, the block line extended further. CONCLUSION: In addition to pacing rate-dependent and site-dependent changes in the block line, pilsicainide further extended the block line length. This phenomenon explains the clinical observation that counterclockwise AFL occurs more frequently than clockwise AFL, and the mechanism of class IC AFL.

Cysteinyl leukotrienes enhance the degranulation of bone marrow-derived mast cells through the autocrine mechanism.

The cysteinyl leukotrienes (LTs), LTC(4), LTD(4), and LTE(4), are potent inflammatory mediators and are involved in allergic reactions, such as bronchoconstriction, eosinophilic inflammation, and allergic cell proliferation. The present study aimed to elucidate the role of constitutively produced cysteinyl LTs in mast cell activation. We used a newly developed quantification method based on mass spectrometry to detect cysteinyl LTs in the cultured medium of mouse bone marrow-derived mast cells (BMMCs), which were obtained by interleukin (IL)-3-conditioned culture of mouse bone marrow. BMMCs were stimulated with immunoglobulin (Ig) E and antigen (IgE/Ag) or lipopolysaccharide for 1 or 24 h. This new quantification method revealed that unstimulated BMMCs produced and secreted LTB4 and LTE4 after 24 h of incubation. The treatment of unstimulated BMMCs for 2 h with montelukast, an antagonist of a cysteinyl LT receptor, CysLT1, resulted in the suppression of a downstream signaling event of this receptor, i.e., the decrease in phosphorylation of extracellular responsive kinases. Thus, cysteinyl LTs constitutively simulate BMMCs through the CysLT1 receptor in an autocrine manner. Treatment of BMMCs for 3 weeks with montelukast, which caused long-term inhibition of the autocrine cyteinyl LT-derived signal, significantly attenuated the IgE/Ag-dependent degranulation, as judged by the decrease in the release of beta-hexosaminidase, an enzyme contained in the granules, whereas the production of cytokines, such as IL-6 and tumor necrosis factor-alpha, were largely unaffected. In conclusion, an autocrine signal derived from constitutively produced cysteinyl LTs predisposes mast cells to the degranulation upon allergic stimulation.

Upper turnover portion of the reentry circuit for typical and reverse typical atrial flutter.

BACKGROUND: The posteromedial right atrium (PMRA) forms a block line during typical atrial flutter (AFL). However, whether upper turnover portion exists at the anterior or posterior superior vena cava (SVC) has not been determined. METHODS: We performed right atrial mapping during AFL in 20 patients (typical AFL, n = 17; reverse typical AFL, n = 3) using an electroanatomical mapping system. RESULTS: Mean AFL cycle length was 224 +/- 20 ms and mean number of mapping points was 140 +/- 27. PMRA formed a block line during both typical and reverse AFL in all patients. However, in 16 of 17 patients mapped with typical AFL, PMRA did not extend superiorly to the orifice of the SVC and AFL wave propagated between the upper limit of the PMRA and the posterior SVC. In the remaining patient mapped with typical AFL, a double potential was recorded along the PMRA continuously between the orifices of the inferior vena cava (IVC) and SVC. In the three patients mapped with reverse typical AFL, a posterior barrier was detected from IVC to the upper limit of the PMRA and AFL wave propagated between the upper limit of the PMRA and the posterior SVC. Mean length from IVC to upper limit of the PMRA was 81 +/- 8% of the length from IVC to SVC. CONCLUSIONS: PMRA forms a functional block line during both typical and reverse typical AFL. The upper turnover portion of reentry circuit for AFL was observed between the upper limit of the PMRA and the posterior SVC in the majority of isthmus-dependent AFL patients.

Generation of new benanomicin analogues by biotransformation using Escherichia coli expressing actinomycete cytochrome P450.

Benanomicins were found as antifungal antibiotics from the culture of an actinomycete with potent antifungal activities in vitro and in vivo. We aimed to generate derivatives superior to benanomicin A by biotransformation using Escherichia coli constructed with bacterial P450 expression system. We found transformation of benanomicin A into two derivatives, 10-hydroxybenanomicin A and 11-O-demethylbenanomicin A by one of the P450-expressed strains which harbored a plasmid carrying a CYP105C1-homologous gene. Unexpectedly, the biotransformed compounds showed weak antifungal activities in vitro compared with those of benanomicin A.

Bioinspired Optical Sensor for Remote Measurement of Small Displacements at a Distance.

Identifying appropriate sites for landing a spacecraft or building permanent structures is critical for extraterrestrial exploration. By tracking the movement of land masses and structures on a planetary surface, scientists can better predict issues that could affect the integrity of the site or structures. A lightweight, low-cost, low-power bioinspired optical sensor is being developed at the National Aeronautics and Space Administration (NASA) Ames Research Center to remotely measure small displacements of land masses on either side of a fault. This paper describes the sensor, which is inspired by the compound eye vision system found in many insects, and the algorithms developed to estimate displacement. The results are presented for indoor and outdoor tests using the sensor to measure the displacement of a specially designed target that is located 0.35, 6, and 30 m from the sensor and is moved 10 mm to the left and right of a centered position, simulating the displacement of land masses on either side of a fault. Measurement uncertainties estimates were a few tenths of a millimeter when the target was located 0.35 and 6 m from the sensor. At the 30 m distance, corrections were required to obtain accuracies in the order of 1 mm.

A novel scheme of dystrophin disruption for the progression of advanced heart failure.

The precise mechanism of the progression of advanced heart failure is unknown. We assessed a new scheme in two heart failure models: (I) congenital dilated cardiomyopathy (DCM) in TO-2 strain hamsters lacking delta-sarcoglycan (SG) gene and (II) administration of a high-dose of isoproterenol, as an acute heart failure in normal rats. In TO-2 hamsters, we followed the time course of the histological, physiological and metabolic the progressions of heart failure to the end stage. Dystrophin localization detected by immunostaining age-dependently to the myoplasm and the in situ sarcolemma fragility evaluated by Evans blue entry was increased in the same cardiomyocytes. Western blotting revealed a limited cleavage of the dystrophin protein at the rod domain, strongly suggesting a contribution of endogenous protease(s). We found a remarkable up-regulation of the amount of calpain-1 and -2, and no change of their counterpart, calpastatin. After supplementing TO-2 hearts with the normal delta-SG gene in vivo, these pathological alterations and the animals' survival improved. Furthermore, dystrophin but not delta-SG was disrupted by a high dose of isoproterenol, translocated from the sarcolemma to the myoplasm and fragmented. These results of heart failure, irrespective of the hereditary or acquired origin, indicate a vicious cycle formed by the increased sarcolemma permeability, preferential activation of calpain over calpastatin, and translocation and cleavage of dystrophin would commonly lead to advanced heart failure.

A novel paradigm for therapeutic basis of advanced heart failure--assessment by gene therapy.

The precise mechanism(s) of the progression of advanced heart failure (HF) should be determined to establish strategies for its treatment or prevention. Based on pathological, molecular, and physiological findings in 3 animal models and human cases, we propose a novel scheme that a vicious cycle formed by increased sarcolemma (SL) permeability, preferential activation of calpain over calpastatin, and translocation and cleavage of dystrophin (Dys) commonly lead to advanced HF. The aim of this article was to assess our recent paradigm that disruption of myocardial Dys is a final common pathway to advanced HF, irrespective of its hereditary or acquired origin, but not intended to provide a comprehensive overview of the various factors that may be involved in the course of HF in different clinical settings. In addition, each component of Dys-associated proteins (DAP) was heterogeneously degraded in vivo and in vitro, i.e. Dys and alpha-sarcoglycan (SG) were markedly destroyed using isolated calpain 2, while delta-SG was not degraded at all. The up-regulation of calpain 2 was confirmed through previously published data that remain insufficient for precise evaluation, supporting our new scheme that the activation of calpain(s) is involved in the steady process of Dys cleavage. In addition, somatic gene therapy is discussed as a potential option to ameliorate the physiological/metabolic indices and to improve the prognosis.

New naphthoquinones, f13102A and B, from a fungus strain F-13102.

Deficiency of Fas-mediated apoptosis is one of the mechanisms involved in the immune evasion by tumors. Thus, it might be a practical approach for cancer treatment that Fas-mediated apoptosis in tumor cells is modified by drugs. In the course of screening, we have isolated two new naphthoquinones, f13102A and B, from the culture broth of fungus strain F-13102. Coumpound f13102A sensitizes Fas-resistant human lung adenocarcinoma A549 cells to apoptosis.

Effect of cytogenin, a novel immunomodulator, on streptozotocin-induced diabetes in mice.

The anti-diabetic effect of cytogenin was examined using streptozotocin-induced diabetes in mice. Cytogenin suppressed not only the increase of plasma glucose level but also the body weight reduction in diabetic mice. Histological examination of the pancreas taken from diabetic mice given cytogenin showed that cytogenin decreased the number of macrophages infiltrated into islet of pancreas. Further, cytogenin suppressed the nitric oxide generation by macrophages treated with lipopolysaccharide through decreasing of inducible nitric oxide synthase expression. Cytogenin suppressed interleukin-6 expression by macrophage treated with LPS, suggesting that the anti-diabetic activity of cytogenin might be partly attributed to the suppressive activity against nitric oxide generation.

A microplate assay for selective measurement of growth of epithelial tumor cells in direct coculture with stromal cells.

Stromal cells play an important role in regulating epithelial malignancies through diffusible factors and adhesion. Modulation of the tumor-stromal cell interaction is an attractive target for new antitumor strategies. To screen for a modulator of the interaction, we have now developed a quantitative colorimetric assay for measurement of tumor cell growth in coculture with stromal cells using rhodanile blue dye. Rhodanile blue specifically stained cytokeratin-positive tumor cells in the coculture. When human prostate carcinoma cells LNCaP, PC-3 and DU-145 were cocultured with normal prostate stromal cells (PrSC) in a microplate, growth of the prostate cancer cells in the coculture was selectively measured by the rhodanile blue staining method. Using this system, we searched for a modulator of the tumor-stromal cell interaction among clinically used drugs and natural products. As a result, we found that 5-fluorouracil, bleomycin and phthoxazolin A inhibit prostate cancer cell growth more strongly in coculture with PrSC than that in monoculture. Without need to pre-label cells and transfect a marker gene, our new method is simple, rapid and thus useful for screening for modulators of the tumor-stromal cell interaction. Furthermore, our results suggest that low molecular weight compounds modulate the tumor-stromal cell interaction.

ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp. F-1490. II. Structure determination and synthesis.

ICM0201 (1), a new inhibitor of murine osteoclastogenesis in culture was isolated from a fermentation broth of Cunninghamella sp. F-1490. The structure of ICM0201 was determined to be (3S,10aR)-3,4a-dihydroxy-2,3,4,4a-tetrahydro-2H-pyrano[3,2-b]benzo[e]morpholine-9-carboxylic acid by spectroscopic analyses and chemical studies. The structure of 1 is unique in that the tricycle ring system is composed of aminal and hemiacetal bonds.

ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. I. Taxonomy, fermentation, isolation and biological activities.

In the course of screening program for inhibitors of angiogenesis, novel substances designated as ICM0301A approximately H (1 approximately 8) were isolated from the culture broth of Aspergillus sp. F-1491. ICM0301s inhibited the growth of human umbilical vein endothelial cells (HUVECs) induced by basic fibroblast growth factor (bFGF) with IC50 values of 2.2 approximately 9.3 microg/ml. ICM0301A (1) showed significant anti-angiogenic activity at lower than 10 microg/ml in the angiogenesis model using rat aorta cultured in fibrin gel. ICM0301s showed very low cytotoxicity against various tumor cells. Furthermore, 1CM0301A did not show any toxic symptom in mice by intraperitoneal injection at 100 mg/kg.

ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. II. Physico-chemical properties and structure elucidation.

ICM0301A (1), B (2) and their congeners (3 approximately 8) were isolated from a culture broth of Aspergillus sp. F-1491 as new angiogenesis inhibitors. Their structures were elucidated by spectroscopic analyses. ICM0301A and B have a substituted decalin skeleton containing two oxirane rings.

ICM0201, a new inhibitor of osteoclastogenesis from Cunninghamella sp. F-1490. I. Taxonomy, fermentation, isolation and biological activities.

In the course of screening for inhibitors of osteoclastogenesis, a new substance designated as ICM0201 was isolated from a fermentation broth of Cunninghamella sp. F-1490. ICM0201 inhibited the formation of osteoclasts in mouse bone marrow cells with an IC50 value of 0.78 microg/ml and showed weak cytotoxicity against bone marrow cells.

Recognition of inferiorly dislocated fast pathways guided by three-dimensional electro-anatomical mapping.

PURPOSE: Slow pathway (SP) ablation of atrioventricular (AV) nodal reentrant tachycardia (AVNRT) can be complicated by unexpected AV block even at sites >10 mm inferior to the bundle of His (HB), and one cause is thought to be the inferior dislocation of an antegrade fast pathway (A-FP). We assessed locations of FPs guided by CARTO. METHODS: Sites of FPs were mapped guided by CARTO before SP ablation in 18 patients with slow-fast AVNRT. The A-FP was defined as the site with the minimum interval between the stimulus and HB potential when pace mapping in the right atrial septum. RESULTS: The A-FP was 7.9 ± 7.5 mm inferior and 2.9 ± 5.0 mm posterior to the HB. In 6 of 18 patients (33%), the A-FP was inferiorly dislocated >10 mm to the HB. SP ablation was successfully performed in all patients at sites >10 mm from both the HB and the A-FP without AV block. In the inferiorly dislocated A-FP group, A-FPs seemed to be positioned much more on atrial sites and sufficiently posterior to SP ablation sites. CONCLUSIONS: The A-FP inferiorly dislocated >10 mm to the HB in one third of patients with AVNRT and seemed to be positioned deep on atrial sites. It is again emphasized that SP ablation within the triangle of Koch should be performed at a very ventricular annulus site, particularly in the inferiorly dislocated A-FP group.

Utility of virtual unipolar electrogram morphologies to detect transverse conduction block and turnaround points of typical atrial flutter.

BACKGROUND: Noncontact mapping is useful for the diagnosis of various arrhythmias. Virtual unipolar electrogram morphologies (VUEM) of the conduction block and the turnaround points, however, are not well defined. We compared the VUEM characteristics of a transverse conduction block in the posterior right atrium (RA) with those of contact bipolar electrograms obtained during typical atrial flutter (AFL). METHODS: Contact bipolar electrograms were used to map the posterior RA during typical AFL in 16 patients. Twenty points of the VUEM recorded along the block line were analyzed and compared with contact bipolar electrograms. RESULTS: Seventeen AFLs were analyzed. Fifteen AFLs showed an incomplete transverse conduction block in the posterior RA by contact bipolar mapping. A double potential on the block line corresponded to the two components of the VUEM, in which the second component showed an Rs, RS, or rS pattern. At the turnaround point, a fused double potential of the contact bipolar electrograms corresponded to a change of the second component of the VUEM from an rS to a QS morphology. Two AFLs showed a complete block line in the posterior RA. The contact bipolar electrogram showed double potentials from the inferior vena cava to the superior vena cava, whereas the second component of the VUEM remained in an unchanged Rs, RS, or rS pattern. CONCLUSION: VUEM analysis was a reliable method for identifying the posterior block line during AFL. This method may also be applicable for detecting block lines and turnaround points of circuits in other unmappable arrhythmias.

Late gadolinium enhancement on cardiac magnetic resonance imaging: is it associated with a higher incidence of nonsustained ventricular tachycardia in patients with idiopathic dilated cardiomyopathy?

PURPOSE: Late gadolinium enhancement (LGE) during cardiac magnetic resonance imaging (MRI) can be seen in patients with myocardial fibrosis accompanied by myocardial infarction and cardiomyopathy. Some idiopathic dilated cardiomyopathy (DCM) patients have fibrosis in the myocardium and show LGE during cardiac MRI. The purpose of this study was to investigate the clinical significance of LGE in patients with DCM. MATERIALS AND METHODS: We recruited 32 DCM patients who had a left ventricular ejection fraction (LVEF) of <40% by echocardiography and performed cardiac MRI. LGE images were obtained 15 min after injection of Gd-DTPA (0.1 mmol/kg) using an inversion recovery gradient echo sequence. We compared LGE(+) and LGE(-) groups in terms of their nonsustained ventricular tachycardia (NSVT) properties. We also compared LGE and the frequency of an implantable cardioverter defibrillator (ICD) implantation. RESULTS: In total, 18 patients (56.3%) had LGE and a higher incidence of NSVT (P = 0.01). ICD implantation was more frequent in the LGE(+) group (P = 0.04). CONCLUSION: Because the LGE(+) patients showed a higher incidence of NSVT and ICD implantation, cardiac MRI could prove to be a useful tool in the management of DCM patients.