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Noninvasive assessment of stenotic lesions in patients with complex adult congenital heart disease (ACHD) is challenging due to its complex morphology. The simultaneous two-screen display of multidetector-computed tomography (MDCT) and real-time echogram (STDME) technology can display a virtual multi-planar reconstruction from MDCT corresponding to the same cross-sectional image from transthoracic echocardiography (TTE). We investigated the usefulness of the STDME technology for stenosis severity assessment in complex ACHD patients.Twenty-four complex ACHD patients with stenotic lesions were enrolled in this study. All patients underwent TTE and the STDME technology within a week after MDCT. Peak velocity and pressure gradient (PG) across the stenotic site were measured using continuous wave Doppler. Cardiac catheterization was performed in 17 patients.Nine out of the twenty-four patients had undergone repair with a conduit. Peak velocity and PG from the STDME technology were higher than those from TTE (peak velocity: 3.1 ± 1.1 vs. 2.8 ± 1.0 m/s; peak PG: 43 ± 28 vs. 34 ± 21 mmHg; both p0.01). Peak PG from the STDME technology showed significant correlations with those from catheterization in patients with a conduit (n=7) and those without a conduit (n=10) (r = 0.795 and 0.880, respectively; both p0.05), while peak PG from TTE was correlated with catheterization measurements only in patients without a conduit (r = 0.850, p0.05).The STDME technology enables more accurate assessment of conduit stenosis severity than does TTE in complex ACHD patients.

Takeuchi repair is a unique surgical approach in anomalous left coronary artery from the pulmonary artery. We present an adult patient with anomalous left coronary artery from the pulmonary artery with multiple late structural complications after Takeuchi repair who was evaluated using multimodality imaging, including newly developed cardiac fusion imaging with computed tomography and echocardiography. (Level of Difficulty: Advanced.), Central Illustration

Few studies have reported the efficacy of Occlutech Figulla Flex II (FFII) device compared with Amplatzer Septal Occluder (ASO) device. The aim of this study was to examine the efficacy and safety of FFII compared with ASO for transcatheter atrial septal defect (ASD) closure. We retrospectively evaluated 190 patients using FFII and 190 patients using ASO who underwent transcatheter ASD closure. ASD characteristics were evaluated by transesophageal echocardiography. The prevalence of procedural complications, including erosion, device embolization, stroke, and new-onset atrial arrhythmia, and the presence of a residual shunt were evaluated between the two groups during 12-month follow-up. FFII was used more frequently than ASO in patients with a deficient aortic rim or septal malalignment (P = 0.02, P

Background Although transthoracic echocardiography (TTE) may be useful for patent foramen ovale (PFO) screening, the optimal methodologies remain unclear. The aims of this study were to evaluate the efficacy of the abdominal compression Valsalva maneuver and identify the optimal cutoff value of microbubbles in contrast TTE for detecting PFO, compared with transesophageal echocardiography and catheterization as the reference. Methods One hundred thirty-four patients with cryptogenic stroke or migraine who had suspected PFO and underwent TTE and transesophageal echocardiography plus catheterization were enrolled. The sensitivity, specificity, and accuracy of TTE for PFO detection were analyzed according to different provocations (spontaneous Valsalva maneuver, abdominal compression Valsalva maneuver) and different cutoff values of microbubbles for a positive result (at least one microbubble, at least five microbubbles). Results Eighty patients had PFO confirmed by transesophageal echocardiography and catheterization. When the cutoff was at least one microbubble, the sensitivity of TTE in detecting PFO was 93% with the spontaneous Valsalva maneuver and 99% with the abdominal compression Valsalva maneuver. When the cutoff was at least five microbubbles, sensitivity was 85% with the spontaneous Valsalva maneuver and 99% with the abdominal compression Valsalva maneuver. With the abdominal compression Valsalva maneuver, specificity was increased using the cutoff of at least five microbubbles compared with at least one microbubble (89% vs 57%). The abdominal compression Valsalva maneuver with the cutoff of at least 5 microbubbles provided the greatest accuracy of 95%. Conclusions TTE with the abdominal compression Valsalva maneuver had excellent sensitivity. The cutoff of at least five microbubbles increased specificity. Our findings suggest that TTE with these criteria is valuable for PFO diagnosis.

Background Transcatheter closure of patent foramen ovale (PFO) has become an effective therapeutic strategy for cryptogenic stroke (CS). The identification of high-risk PFO is essential, but the data are limited. This study aimed to clarify the factors related to CS and to develop a score for high-risk PFO. Methods We retrospectively analyzed 57 patients with prior CS and 50 without CS who were scheduled for transcatheter closure. PFO characteristics were evaluated by transesophageal echocardiography. Based on factors related to CS, we estimated the risk score. Results Patients with CS had a greater frequency of large-size PFO (≥2 mm in height), long-tunnel PFO (≥10 mm in length), atrial septal aneurysm, hypermobile interatrial septum, prominent Eustachian valve or Chiari's network, the large right-to-left shunt at rest and during Valsalva maneuver, and low-angle PFO (≤10° of PFO angle from inferior vena cava), compared with patients without CS. Multivariate analysis showed that long-tunnel PFO, the presence of hypermobile interatrial septum, the presence of prominent Eustachian valve or Chiari's network, the large right-to-left shunt during Valsalva maneuver, and low-angle PFO were independently related to CS. When the score was estimated based on 1 point for each factor, the proportion of CS was markedly elevated with a score of ≥2 points. The probability of CS was markedly different between scores of ≤1 or ≥2 points. Conclusions PFO risk can be assessed with a score based on high-risk features. The presence of two or more high-risk PFO features is associated with CS.

Objective. The aim of this study was to examine the relationship between right ventricular (RV) volume and exercise capacity in adult patients with atrial septal defect (ASD) and to determine the degree of RV dilatation for transcatheter ASD closure. Background. RV dilatation is an indication of transcatheter ASD closure; however, few studies have reported the clinical significance of RV dilatation. Methods. We enrolled 82 consecutive patients (mean age, 49 ± 18 years; female, 68%) who underwent cardiac magnetic resonance imaging and symptom-limited cardiopulmonary exercise test before ASD closure. The relationship between RV volume and peak oxygen uptake (VO2) was evaluated. Results. The mean RV end-diastolic volume index was 108 ± 27 ml/m2 (range, 46 to 180 ml/m2). The mean peak VO2 was 24 ± 7 ml/min/kg (range, 14 to 48 ml/min/kg), and the mean predicted peak VO2 was 90 ± 23%. There were significant negative relationships of RV end-diastolic volume index with peak VO2 (r = −0.28, p<0.01) and predicted peak VO2 (r = −0.29, p<0.01). The cutoff value of RV end-diastolic volume index 2 was 120 ml/m2, with the sensitivity of 49% and the specificity of 89%. Conclusions. There was a relationship between RV dilatation and exercise capacity in adult patients with ASD. RV end-diastolic volume index ≥120 ml/m2 was related to the reduction in peak VO2. This criterion of RV dilatation may be valuable for the indication of transcatheter ASD closure.

Background. Septal malalignment is related to erosion and device embolization in transcatheter closure of atrial septal defect (ASD), but limited information is available. Objectives. This study aimed to assess clinical significance of septal malalignment and to determine appropriate evaluation of ASD diameter, including the selection of device size. Methods. Four hundred and seventeen patients with ASD who underwent transcatheter closure were enrolled. Septal malalignment was defined as separation between the septum primum and the septum secundum on transesophageal echocardiography. Results. One hundred and eighty-four patients had septal malalignment. The frequency of septal malalignment increased with age reaching around 50% in adult patients. Septal malalignment was related to aortic rim deficiency. The distance of separation between the septum primum and the septum secundum was 5 ± 2 mm (range, 1–11 mm). In patients with septal malalignment, the ASD diameter measured at the septum primum was 19 ± 6 mm, while the ASD diameter measured at the septum secundum was 16 ± 6 mm. There was a difference of 4 ± 2 mm (range, 0–8 mm) between the ASD diameter measured at the septum primum and that measured at the septum secundum. For transcatheter closure, the Amplatzer Septal Occluder device size 2-3 mm larger and the Occlutech Figulla Flex II device size 4–7 mm larger than the ASD diameter measured at the septum primum were frequently used. During the study period, erosion or device embolization did not occur in all of the patients. Conclusions. Septal malalignment is highly prevalent in adult patients with aortic rim deficiency. The measurement of ASD diameter at the septum primum can be valuable for the selection of device size in patients with septal malalignment.

A high-pressure crystallographic study was conducted on Escherichia coli dihydrofolate reductase (ecDHFR) complexed with folate and NADP+ in crystal forms containing both the open and closed conformations of the M20 loop under high-pressure conditions of up to 800 MPa. At pressures between 270 and 500 MPa the crystal form containing the open conformation exhibited a phase transition from P21 to C2. Several structural changes in ecDHFR were observed at high pressure that were also accompanied by structural changes in the NADP+ cofactor and the hydration structure. In the crystal form with the closed conformation the M20 loop moved as the pressure changed, with accompanying conformational changes around the active site, including NADP+ and folate. These movements were consistent with the suggested hypothesis that movement of the M20 loop was necessary for ecDHFR to catalyze the reaction. In the crystal form with the open conformation the nicotinamide ring of the NADP+ cofactor undergoes a large flip as an intermediate step in the reaction, despite being in a crystalline state. Furthermore, observation of the water molecules between Arg57 and folate elucidated an early step in the substrate-binding pathway. These results demonstrate the possibility of using high-pressure protein crystallography as a method to capture high-energy substates or transient structures related to the protein reaction cycle.

Background Management of adult congenital heart disease (ACHD) patients requires understanding of its complex morphology and functional features. An innovative imaging technique has been developed to display a virtual multi-planar reconstruction obtained from contrast-enhanced multidetector-computed tomography (MDCT) corresponding to the same cross-sectional image from transthoracic echocardiography (TTE). The aim of this study is to assess the usefulness of this imaging technology in ACHD patients. Methods This study consisted of 46 consecutive patients (30 women; mean age, 52 ± 18 years old) with ACHD who had undergone contrast MDCT. All patients underwent TTE within a week of MDCT. An experienced sonographer who did not know the results of MDCT conducted a diagnosis using TTE and, then, using the new imaging technology. We studied whether this imaging technology provided additional or unexpected findings or makes more accurate diagnosis. Results In this imaging technology, MDCT cross-section provides higher-resolution image to the deep compared to corresponding TTE image. Depending on the MDCT section which can be arbitrarily set under the echo guide, we can diagnose unexpected or incremental lesions or more accurately assess the severity of the lesion in 27 patients (59%) compared to TTE study alone. This imaging technology was useful in the following situations: 1. For anatomical guidance for Doppler flow assessment. 2. Assessment of morphology of obstruction in outflow tracts or conduit. 3. To clarify the etiology of unordinary severe valvular disease. 4. Assessment of the patient with diseased prosthetic valves. Conclusions This integrated imaging technology provides incremental role over TTE in complex anatomy, and allows functional information in ACHD patients.

Waste disposal site is one of the important sinks of chemicals. A significant amount of perfluoroalkyl and polyfluoroalkyl substances (PFASs) such as perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS), and perfluorohexanoic acid (PFHxA) have been brought into it. Because of their aqueous solubility, PFASs are released to landfill effluent waters, from which PFASs are efficiently collected by adsorption technique using granular activated carbon (GAC). The exhausted GAC is reactivated by heating processes. The mineralization of PFASs during the reactivation process was studied. Being thermally treated in N2 atmosphere, the recovery rate of mineralized fluorine and PFC homologues including short-chained perfluorocarboxylic acids was determined. If the reagent form of PFOA, PFHxA, and PFOS were treated at 700 °C, the recovery of mineralized fluorine was less than 30, 46, and 72 %, respectively. The rate increased to 51, 74, and 70 %, if PFASs were adsorbed onto GAC in advance; moreover, addition of excess sodium hydroxide (NaOH) improved the recovery to 74, 91, and 90 %. Residual PFAS homologue was less than 1 % of the original amount. Steamed condition did not affect destruction. The significant role of GAC was to suppress volatile release of PFASs from thermal ambient, whereas NaOH enhanced destruction and retained mineralized fluorine on the GAC surface. Comparing the recovery of mineralized fluorine, the degradability of PFOS was considered to be higher than PFOA and PFHxA. Whole mass balance missing 9~26 % of initial amount suggested formation of some volatile organofluoro compounds beyond analytical coverage.

A 73-year-old woman presented with fever and Janeway lesions on her palms. She had undergone transcatheter patent foramen ovale closure with a 30-mm Amplatzer Cribriform device (Abbott, Chicago, Illinois) 6 years before. Two months before onset, she had received dental procedures without antibiotic

Objective This study evaluated the effect of echocardiographic left ventricular (LV) diastolic dysfunction on acute congestive heart failure after transcatheter atrial septal defect (ASD) closure in elderly patients. Background Although there is concern that LV diastolic dysfunction develops acute congestive heart failure after ASD closure, limited information is available regarding the influence, especially in elderly patients with severe LV diastolic dysfunction. Methods Two hundred consecutive patients older than 60 years were divided into 3 groups according to echocardiographic LV diastolic dysfunction: severe (early diastolic mitral annular velocity [e’]

The aim of this study was to demonstrate the feasibility and usefulness of addition of the right parasternal approach to the conventional left parasternal and apical approaches using two-dimensional (2D) and three-dimensional (3D) transthoracic echocardiography (TTE) for morphologic evaluation in cases of transcatheter closure of atrial septal defects (ASDs).In 112 consecutive patients with ASDs, the morphology of the defects was evaluated for transcatheter closure in the right parasternal view in addition to the conventional left views using 2D and 3D TTE. Measurements of the maximal ASD diameter and detection of deficient rim obtained on 2D TTE were compared with those obtained by 2D transesophageal echocardiography. The shapes and locations of ASDs visualized by 3D TTE were compared with those visualized by 3D transesophageal echocardiography.In 88 patients (80.0%), optimal images from the right parasternal approach for morphologic evaluation of ASDs were obtained. Although there was a significant difference in maximal ASD diameter obtained only in the conventional left approach compared with transesophageal echocardiographic measurements (P.05), when the right parasternal approach was applied, a significant difference was not found (P = .18), and the diagnostic concordance of the rim deficiency was improved from 85.2% to 90.9%. Three-dimensional TTE from the right parasternal approach improved visualization of the shape and location of ASDs from 65.5% to 74.5%.Additional use of the right parasternal approach enables detailed morphologic evaluation for transcatheter closure of ASDs. In patients with suboptimal images on 3D TTE in the left conventional approach, additional 3D TTE in the right parasternal approach can improve the feasibility of obtaining optimal 3D images to evaluate the shapes and locations of ASDs.

Structures of 3-­isopropylmalate dehydrogenase were determined at pressures ranging from 0.1 to 650 MPa. Comparison of these structures gives a detailed picture of the swelling of a cavity at the dimer interface and the generation of a new cleft on the molecular surface, which are accompanied by water penetration., Hydrostatic pressure induces structural changes in proteins, including denaturation, the mechanism of which has been attributed to water penetration into the protein interior. In this study, structures of 3-isopropylmalate dehydrogenase (IPMDH) from Shewanella oneidensis MR-1 were determined at about 2 Å resolution under pressures ranging from 0.1 to 650 MPa using a diamond anvil cell (DAC). Although most of the protein cavities are monotonically compressed as the pressure increases, the volume of one particular cavity at the dimer interface increases at pressures over 340 MPa. In parallel with this volume increase, water penetration into the cavity could be observed at pressures over 410 MPa. In addition, the generation of a new cleft on the molecular surface accompanied by water penetration could also be observed at pressures over 580 MPa. These water-penetration phenomena are considered to be initial steps in the pressure-denaturation process of IPMDH.

Background The aim of this study was to demonstrate the utility of real-time three-dimensional (3D) transesophageal echocardiography (RT3D-TEE) using a matrix array 3D transesophageal echocardiographic probe for morphologic evaluation and guidance of transcatheter closure of atrial septal defects (ASDs). Methods Forty-eight consecutive patients scheduled for the intervention were included. Two-dimensional (2D) transesophageal echocardiography (2D-TEE) and RT3D-TEE were performed before and during the procedures. Measurements of maximal ASD diameter and surrounding rims obtained on RT3D-TEE were compared with those obtained on 2D-TEE. Results In 46 patients (96%), optimal 3D images for the morphologic evaluation of ASDs were obtained. RT3D-TEE facilitated the evaluation of ASD morphology and surrounding rims and was able to provide intraprocedural information clearly. A Bland-Altman plot showed a mean maximal diameter difference of −0.12 mm between the means (95% limits of agreement, −2.2 to 2.5 mm). Conclusion RT3D-TEE is a clinically useful, complementary option to 2D-TEE for evaluation of ASD morphology and for interventional guidance.

To elucidate the heme acquisition system in pathogenic bacteria, we investigated the heme-binding properties of the third NEAT domain of IsdH (IsdH-NEAT3), a receptor for heme located on the surfaces of pathogenic bacterial cells, by using x-ray crystallography, isothermal titration calorimetry, examination of absorbance spectra, mutation analysis, size-exclusion chromatography, and analytical ultracentrifugation. We found the following: 1) IsdH-NEAT3 can bind with multiple heme molecules by two modes; 2) heme was bound at the surface of IsdH-NEAT3; 3) candidate residues proposed from the crystal structure were not essential for binding with heme; and 4) IsdH-NEAT3 was associated into a multimeric heme complex by the addition of excess heme. From these observations, we propose a heme-binding mechanism for IsdH-NEAT3 that involves multimerization and discuss the biological importance of this mechanism.

A 38-year-old man was referred to our hospital because of progressive dyspnea on exertion. He was diagnosed as having congestive heart failure. On physical examination, his blood pressure was 118/86 mm Hg, and his heart rate was regular at 80 beats/min. On admission, jugular venous distension was detected, a third heat sound was audible, and auscultation of the lungs revealed bilateral coarse crackles and diminished breath sounds. Peripheral edema was also detected. A chest x-ray film showed a marked cardiac silhouette, with right-sided pleural effusion (Figure 1A). ECG on admission showed sinus rhythm with poor R progression and ST depression in aVF and V6 leads (Figure 1B). A blood test showed elevated plasma brain natriuretic peptide (417.9 pg/mL) and d-dimer (6.7 μg/m) levels. Transthoracic echocardiography revealed normal-sized left and right ventricles and the presence of a small amount of pericardial effusion; the left ventricular ejection fraction was severely decreased. The left and right atria were markedly …

The HIV-1 Vif protein inactivates the cellular antiviral cytidine deaminase APOBEC3F (A3F) in virus-infected cells by specifically targeting it for proteasomal degradation. Several studies identified Vif sequence motifs involved in A3F interaction, whereas a Vif-binding A3F interface was proposed based on our analysis of highly similar APOBEC3C (A3C). However, the structural mechanism of specific Vif-A3F recognition is still poorly understood. Here we report structural features of interaction interfaces for both HIV-1 Vif and A3F molecules. Alanine-scanning analysis of Vif revealed that six residues located within the conserved Vif F1-, F2-, and F3-box motifs are essential for both A3C and A3F degradation, and an additional four residues are uniquely required for A3F degradation. Modeling of the Vif structure on an HIV-1 Vif crystal structure revealed that three discontinuous flexible loops of Vif F1-, F2-, and F3-box motifs sterically cluster to form a flexible A3F interaction interface, which represents hydrophobic and positively charged surfaces. We found that the basic Vif interface patch (R17, E171, and R173) involved in the interactions with A3C and A3F differs. Furthermore, our crystal structure determination and extensive mutational analysis of the A3F C-terminal domain demonstrated that the A3F interface includes a unique acidic stretch (L291, A292, R293, and E324) crucial for Vif interaction, suggesting additional electrostatic complementarity to the Vif interface compared with the A3C interface. Taken together, these findings provide structural insights into the A3F-Vif interaction mechanism, which will provide an important basis for development of novel anti-HIV-1 drugs using cellular cytidine deaminases. IMPORTANCE HIV-1 Vif targets cellular antiviral APOBEC3F (A3F) enzyme for degradation. However, the details on the structural mechanism for specific A3F recognition remain unclear. This study reports structural features of interaction interfaces for both HIV-1 Vif and A3F molecules. Three discontinuous sequence motifs of Vif, F1, F2, and F3 boxes, assemble to form an A3F interaction interface. In addition, we determined a crystal structure of the wild-type A3F C-terminal domain responsible for the Vif interaction. These results demonstrated that both electrostatic and hydrophobic interactions are the key force driving Vif-A3F binding and that the Vif-A3F interfaces are larger than the Vif-A3C interfaces. These findings will allow us to determine the configurations of the Vif-A3F complex and to construct a structural model of the complex, which will provide an important basis for inhibitor development.

Introduction: In patients with aortic stenosis (AS) and atrial fibrillation (AF), the conventional continuity equation to evaluate aortic valve area (AVA) is cumbersome, because 5 to 10 cycles are required to ensure accuracy of results. Double-envelope (DE) is obtained by a single continuous-wave Doppler envelope with double density velocity profiles; the inner envelope represents flow across the left ventricular outflow tract and outer envelope represents flow across the aortic valve orifice. Hypothesis: The aim of this study to evaluate the usefulness of the AVA calculated from single-beat recorded inner and outer envelopes in patients with AS and AF. Methods: Eighteen AS patients (74.6±7.2 years old) with AF were examined by transthoracic echocardiography. The conventional AVA (PW/CW technique) was calculated from nonsimultaneously measured LVOT flow and AVO flow in randomly picked up 8 cardiac cycles, respectively. The AVA (DE technique) was calculated from the inner and outer envelopes simultaneously recorded. A single-beat AVA was calculated from simultaneously recorded inner and outer envelopes (by DE technique) when the preceding RR interval/pre-preceding RR interval = 1. Bland-Altman analysis was used to compare the results of two techniques and estimate the reproducibility. Results: DE profiles were successfully obtained in all patients, and mean AVA (PW/CW technique) was 1.09±0.19 cm2. AVA by Single-beat DE technique showed good correlation with that by PW/CW technique (r=0.90), and the mean bias in the AVA measurements between by PW/CW technique and by DE technique was 0.048 cm2. (Figure) Conclusions: AVA in patients with AS and AF obtained by Single-beat DE technique was feasible and in good agreement with that by PW/CW technique. We suggest that DE technique should be considered to estimate the subtle temporal change of AVA in patient with AS and AF more simply and accurately.

Hepatocyte nuclear factor 6 (HNF-6)/OC-1, a part of liver-enriched transcription factor, controls pancreas and liver development and regulates expression of several hepatic genes. DNA-binding region of HNF-6alpha bound to a 14-mer DNA fragment has been crystallized by the hanging drop vapor diffusion method. The crystals belong to space group P2 with unit cell parameters of a = 73.0 A, b = 39.0 A, c = 106.5 A, beta = 107.6 degrees. X-ray diffraction data were collected to 2.0 A resolution.

MRP14 is a protein that is specifically expressed in myeloid and epithelial cells during the stages of acute or chronic inflammatory states such as rheumatoid arthritis or sarcoidosis. MRP14 has EF-hand motifs as Ca(2+)-binding sites and belongs to the S100 family of proteins. This paper deals with the sample preparation (cloning, overexpression and purification), crystallization and preliminary crystallographic analysis of recombinant human MRP14. Crystals of MRP14 were obtained by the hanging-drop vapour-diffusion method. MRP14 crystals belong to space group P2(1), with unit-cell parameters a = 57.59, b = 178.44, c = 61.23 A, beta = 113.17 degrees, and diffract to 2.1 A resolution.

We have investigated the pharmacology of sodium (R)-2-[4-(2,2-dimethyl-1,3-dioxan-5-yl) methoxy-3,5-dimethylpyridin-2-yl]methylsulfinyl-1H-benzimidazol (E3710), a new proton pump inhibitor (PPI), and its effect on gastric acid secretion. E3710 irreversibly inhibited H(+),K(+)-ATPase activity in pig gastric vesicles with an acidic internal environment with an IC(50) of 0.28 microM. Administration of E3710 (0.1, 0.2, 0.4, and 0.8 mg/kg; n = 6) intraduodenally in a gastric fistula model in dogs inhibited histamine-stimulated gastric acid secretion at 0 to 2 and 24 to 26 h after administration with ED(50) values of 0.18 and 0.22 mg/kg, respectively. The inhibition by E3710 was 2.3 times more potent than that of another representative PPI, esomeprazole (0.2, 0.4, 0.8, and 1.6 mg/kg; n = 6) at 0 to 2 h after administration (ED(50) = 0.40 mg/kg) and 2.8 times more potent at 24 to 26 h (ED(50) = 0.71 mg/kg). In the gastric fistula dogs, the intragastric pH wasor=4 for 17% (n = 27) of a 24-h period with vehicle alone, but when E3710 was administered, at 0.2 (n = 4), 0.4 (n = 8), and 0.8 mg/kg (n = 5), the pH wasor=4 for 40, 79, and 88% of a day, respectively. The corresponding values for esomeprazole at 0.8 (n = 4) and 1.6 mg/kg (n = 8) were 55 and 59%, respectively. In a crossover study with vehicle, E3710 at 0.4 mg/kg and esomeprazole at 1.6 mg/kg (n = 6), E3710 increased the intragastric pH to4 for 82% of a day compared with 61% of a day with esomeprazole. These results show that E3710 is a long-acting inhibitor of gastric acid secretion and a promising novel therapy for acid-related diseases, such as gastroesophageal reflux disease.

In ruminants, some leaf-eating animals, and some insects, defensive lysozymes have been adapted to become digestive enzymes, in order to digest bacteria in the stomach. Digestive lysozyme has been reported to be resistant to protease and to have optimal activity at acidic pH. The structural basis of the adaptation providing persistence of lytic activity under severe gastric conditions remains unclear. In this investigation, we obtained the crystallographic structure of recombinant bovine stomach lysozyme 2 (BSL2). Our denaturant and thermal unfolding experiments revealed that BSL2 has high conformational stability at acidic pH. The high stability in acidic solution could be related to pepsin resistance, which has been previously reported for BSL2. The crystal structure of BSL2 suggested that negatively charged surfaces, a shortened loop and salt bridges could provide structural stability, and thus resistance to pepsin. It is likely that BSL2 loses lytic activity at neutral pH because of adaptations to resist pepsin.

Objectives: The aim of this study is to evaluate the feasibility and efficacy of device closure of atrial septal defect (ASD) in elderly patients with permanent atrial fibrillation. Background: Little is known about the feasibility of device closure of ASD in those patients. Methods: Nine consecutive patients (mean age 68.1 years) with permanent atrial fibrillation (>1 year persistent) underwent catheter closure using the Amplatzer septal occluder. Transthoracic echocardiography and plasma B-type natriuretic peptide (BNP) level were assessed before and at 24 hours; and 1, 3, and >6 months after the closure. Before the procedure, appropriate dose of warfarin was used in all, diuretics was used in 8/9. Same amount of medications were continued after the procedure. Results: ASD could be closed in all (mean device size 27.3 mm) without hemodynamic and thromboembolic complications. New York Heart Association (NYHA) functional classification was significantly improved in all patients after device closure. No hemodynamic and thromboembolic complications were observed during the follow-up period (mean 10.6 months). Although permanent atrial fibrillation did not change in all after the procedure, resting heart rate decreased from 76.2 ± 16.0 to 68.3 ± 13.2 beats/min (P = 0.015). There was statistically significant improvement in right ventricular/left ventricular diameter ratio (1.08 ± 0.16 to 0.73 ± 0.10, P = 0.008) and plasma BNP level (183.7 ± 90.5 to 94.6 ± 47.4 pg/mL, P = 0.008) after >6 months device closure. Conclusions: Even in the patients complicated with permanent fibrillation, transcatheter closure of ASD can contribute to symptomatic improvement as well as cardiac geometric remodeling. © 2009 Wiley-Liss, Inc.

Background: Wave intensity (WI) is a new hemodynamic index that provides information about the dynamic behavior of the heart and the vascular system and their interaction. WI can be defined at any site in the circulatory system and provide a great deal of information. However, the clinical usefulness of the second peak of WI, which is related to the ability of the left ventricle to activity stop aortic blood flow, has not been fully reported. In previous study, the magnitude of W2 was related to the inertia force of late systolic aortic flow. Also, the higher augmented inertia force, which was calculated with cardiac catheterization, was associated with greater exercise capacity. Then, we hypothesize that WI is correlated to exercise capacity. In this study, we investigated whether WI and the echocardiographic parameters would be correlated with exercise capacity. Methods: A total of 24 patients of heart disease were enrolled. WI was obtained at the right carotid artery using a color Doppler system for blood velocity measurement combined with an echo-tracking method for detecting vessel diameter changes. As echocardiographic parameters, we analyzed ejection fraction (EF), the early (E) and late (A) diastolic mitral velocities, and early (Ea) and late (Aa) diastolic velocities measured using Tissue Doppler imaging. We evaluated exercise capacity (peak VO2, VE/VCO2, and AT). We used Pearson’s correlation analysis for statistics. Results: EF, the E/A ratio, and the E/Ea ratio were not correlated to exercise capacity. However, the second peak of WI was significant correlated to the peak VO2 (r=0.50, p

Wild type of bovine thrombin has been crystallized in a ligand-free form by the hanging drop vapor diffusion method with polyethylene glycol 4000 and 2-propanol. The crystals belong to space group P4 3 2 12 with unit cell parameters of a = b = 87.7 A, c = 195.9 A. X-ray diffraction data were collected to 2.8 A resolution.

SummaryThe 1.1 MDa cell-wall-associated adhesion protein of staphylococci, Ebh, consists of several distinct regions, including a large central region with 52 imperfect repeats of 126 amino acid residues. We investigated the structure of this giant molecule by X-ray crystallography, circular dichroism (CD) spectrometry, and small-angle X-ray scattering (SAXS). The crystal structure of two repeats showed that each repeat consists of two distinct three-helix bundles, and two such repeats are connected along the long axis, resulting in a rod-like structure that is 120 Å in length. CD and SAXS analyses of the samples with longer repeats suggested that each repeat has an identical structure, and that such repeats are connected tandemly to form a rod-like structure in solution, the length of which increased proportionately with the number of repeating units. On the basis of these results, it was proposed that Ebh is a 320 nm rod-like molecule with some plasticity at module junctions.

Amyloid fibril formation is associated with several pathologies, including Alzheimer's disease, Parkinson's disease, type II diabetes, and prion diseases. Recently, a relationship between basement membrane components and amyloid deposits has been reported. The basement membrane protein, laminin, may be involved in amyloid-related diseases, since laminin is present in amyloid plaques in Alzheimer's disease and binds to amyloid precursor protein. Recently, we showed that peptide A208 (AASIKVAVSADR), the IKVAV-containing peptide, formed amyloid-like fibrils. We previously identified 60 cell adhesive sequences in laminin-1 using a total of 673 12-mer synthetic peptides. Here, we screened for additional amyloidogenic sequences among 60 cell adhesive peptides derived from laminin-1. We first examined amyloid-like fibril formation by the 60 active peptides with Congo red, a histological dye binding to many amyloid-like proteins. Thirteen peptides were stained with Congo red. Four of the 13 peptides promoted cell attachment and neurite outgrowth like the IKVAV-containing peptide. The four peptides also showed amyloid-like fibril formation in both X-ray diffraction and electron microscopic analyses. The amyloidogenic peptides contain consensus amino acid components, including both basic and acidic amino acids and Ser and Ile residues. These results indicate that at least five laminin-derived peptides can form amyloid-like fibrils. We conclude that the laminin-derived amyloidogenic peptides have the potential to form amyloid-like fibrils in vivo, possibly when laminin-1 is degraded.

H+ -transporting ATP synthase (H+ -ATPase) is a multi-subunit complex which acts to produce ATP molecules. The catalytic subunit A of the archaeal-type H+ -ATPase from Pyrococcus horikoshii OT3 was cloned, expressed in Escherichia coli, purified and crystallized by the hanging-drop vapour-diffusion method with MPD as a precipitant. X-ray intensity data were collected to 2.55 A resolution at beamline BL41XU of SPring-8. The crystals belong to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 128.0, c = 104.7 A, and contain one molecule per asymmetric unit.

A 29-year-old man was hospitalized because of dyspnea and palpitation. Electrocardiography demonstrated normal sinus rhythm and borderline right atrial enlargement. Twodimensional transthoracic echocardiography (2D-TTE) revealed a normal left ventricular systolic function. There was a paradoxical septal motion consistent with right ventricular volume overload (Fig. 1a). Although the right ventricle was mildly dilated, its systolic function was preserved. 2D-TTE revealed a large atrium without distinct interatrial septum (Fig. 1b). Left-to-right shunt flow was not detected clearly through the defect on color-flow Doppler imaging. Real-time three-dimensional echocardiography (RT3D-TTE) visualized a large defect of interatrial septum clearly (Fig. 2). The patient was referred for surgery. Only a tiny vestige of the interatrial septum could be confirmed with the naked eye, and the two atrioventricular valves were normal. The patch closure was performed successfully using a fresh autologous pericardial patch. Atrial septal defect (ASD) is a common congenital heart disease presenting in adulthood. Our patient showed a large secundum ASD like single atrium. It is sometimes difficult to distinguish a large secundum ASD from a single atrium. Levy et al. recommended that the term ‘‘single atrium’’ should be used to denote a condition comprising: (1) complete absence of the interatrial septum, (2) absence of malformation of the atrioventricular valves, and (3) absence of interventricular communication [1]. In our case, the morphological diagnosis was supposed to be a large secundum type atrial septal defect, because a small part of the interatrial septum was found. In this case, RT3D-TTE enhanced the understanding of the anatomy and spatial relationship between the defect and its adjacent structure. Previous study has shown the usefulness of RT3D-TTE for patient selection for surgical or transcatheter closure of atrial septal defects. It has been reported that RT3D-TTE allows accurate determination of the ASD location, ASD size, and surrounding tissue of the interatrial septum [2, 3]. Although our patient demonstrated a large part to be missing from the interatrial septum, RT3D-TTE could clearly visualize a large defect of the interatrial septum. RT3D-TTE provided additional, clinically relevant qualitative information for the evaluation of this patient with a large defect of the interatrial septum. N. Watanabe Y. Tanabe Division of Medical Support, Okayama University Hospital, Okayama, Japan

Background: Little is known about right ventricular (RV) dysfunction in patient with pulmonary arterial hypertension (PAH). Recently, RV ischemia has been reported to be a contributory factor in the development of RV dysfunction. The aim of this study is to evaluate RV myocardial perfusion and assess its relationship with RV dysfunction. Methods: This prospective study consisted of 10 consecutive patients (7 female, 51±17 years) with PAH (6 idiopathic PAH (IPAH), and 4 chronic thromboembolic pulmonary hypertension (CTEPH)), and 6 healthy controls (3 females, 37±6 years). Quantitative myocardial contrast echocardiography (MCE) was performed using Philips iE33 imaging system with an S5-1 probe. We depicted the apical four chamber view. A solution of Levovist (300mg/ml) was inravenously administered at a rate of 3ml/min with a volumetric pump. End-systolic images were obtained every four heart beats. The MCE images were analyzed off-line using VoluMap-445 system. The RV myocardium was divided into six segments, at each myocardial segment, calibrated contrast intensity (CI) was determined as the difference (dB) between the mean myocardial CI and cavity CI. 2D speckle tracking echocardiographic study was also performed using Vivid E9 ultrasound system. Results: Among 96 RV myocardial segments, quantitative CI analysis was possible in 86 segments (90%). RV fractional area change was 46% (healthy), 34% (CTEPH) and 25% (IPAH). Mean calibrated CI of RV free wall was -13.4dB, -19.3dB and -22.8dB, respectively (p=0.01). We found good inverse correlation between calibrated CI and longitudinal strain of RV free wall (r=-0.5, p=0.0013). ![Figure][1] Conclusion: Quantitative MCE revealed patients with PAH have impaired RV myocardial perfusion, and this correlated well with the RV longitudinal myocardial dysfunction. [1]: pending:yes

Desmosomes are intercellular junctions, particularly characteristic of epithelial cells, and they are major contributors to cell-cell adhesion in many epithelial cells. Decreased intercellular adhesion has been reported for many carcinomas, and it has been suggested that in part, such alterations in adhesion may be due to changes in adhering junctions, such as the desmosome [1]. In the human urinary bladder, a correlation between decreased numbers of desmosomes and aggressiveness of transitional cell carcinomas has been reported [2]. To define highly malignant esophageal squamous cell carcinoma (SCC) we have investigated the relationship of desmosome numbers to tumor invasiveness and metastasis in esophageal SCCs in a quantitative electron microscopic study [3]. We have also studied this structure with reference to outcome, because all cases studied had been followed for at least 5 years.

MinD is one of the proteins regulating cell division. MinD from Escherichia coli has been designated as a type of motor protein which has an ATPase activity. This paper deals with the first crystallization and preliminary crystallographic analysis of recombinant MinD from Pyrococcus horikoshii (molecular weight 26.3 kDa) expressed in E. coli. Crystals of MinD were obtained by the hanging-drop vapour-diffusion method. MinD crystals belong to space group P2(1)3, with unit-cell parameters a = b = c = 98.5 A, and diffract to 3.0 A resolution. The asymmetric units each contain one molecule of MinD, giving a crystal volume per protein mass (V(M)) of 3.0 A(3) Da(-1) and a solvent content of 59.0%.

C-type lysozyme catalyzes the hydrolysis of peptidoglycan of bacterial cell wall. Most of C-type lysozymes express to protect body against bacterial infection. However, ruminants and some leaf-eating animals have evolved their lysozyme as digestive enzyme. They recruit bacteria which ferment cellulose in the foregut, and digest the bacteria by lysozyme in the true stomach to obtain nutrient. Digestive lysozyme has acquired some properties, such as low optimal pH, and resistance to protease and acid hydrolysis. The structural basis for these properties still remains unclear. In this investigation, we have obtained the crystallographic structure of bovine stomach lysozyme (BSL). This is the first report on the structure of vertebrate digestive lysozyme. We have carried out the denaturant-unfolding experiment and revealed that BSL has high structural stability at acidic pH compared to non-digestive (hen egg-white) lysozyme. The structural stability in acidic solution would be related to the pepsin resistance which was previously reported for BSL. The crystal structure of BSL suggested that negatively charged surfaces, a shortened loop, and salt bridges should provide the structural stability.