Prediction of cell cycle distribution after drug exposure by high content imaging analysis using low-toxic DNA staining dye.

Interference in cell cycle progression has been noted as one of the important properties of anticancer drugs. In this study, we developed the cell cycle prediction model using high-content imaging data of recipient cells after drug exposure and DNA-staining with a low-toxic DNA dye, SiR-DNA. For this purpose, we exploited HeLa and MCF7 cells introduced with a fluorescent ubiquitination-based cell cycle indicator (Fucci). Fucci-expressing cancer cells were subjected to high-content imaging analysis using OperettaCLS after 36-h exposure to anticancer drugs; the nuclei were segmented, and the morphological and intensity properties of each nucleus characterized by SiR-DNA staining were calculated using imaging analysis software, Harmony. For the use of training, we classified cells into each phase of the cell cycle using the Fucci system. Training data (n = 7500) and validation data (n = 2500) were randomly sampled and the binary classification prediction models for G1, early S, and S/G2/M phases of the cell cycle were developed using four supervised machine learning algorithms. We selected random forest as the model with the best performance through 10-fold cross-validation; the accuracy rate was approximately 75%-87%. Regarding feature importance, variables expected to be biologically related to the cell cycle, for example, signal intensity and nuclear size, were highly ranked, suggesting the validity of the model. These results showed that the cell cycle can be predicted in cancer cells by simply exploiting the current prediction model using fluorescent images of DNA-staining dye, and the model could be applied for the use of future ex vivo drug sensitivity diagnosis.

Decomposition of the anisotropic strain in 3D-structure GaN layers using Raman spectroscopy.

Strain engineering for gallium nitride has been studied by many researchers to improve the performance of various devices (i.e., light-emitting diodes, laser diodes, power devices, high electron mobility transistors, and so on). Further miniaturization of gallium nitride devices will clearly continue in the future, and therefore an accurate understanding of the strain state in the devices is essential. However, a measurement technique for axially resolved evaluation of the strain in microareas has not yet been established. In this study, we revealed that the anisotropic strain state induced in c-plane growth gallium nitride is linked to the split state of Raman peaks, which were measured with [Formula: see text] and [Formula: see text] polarized configurations. The anisotropic strain state in c-plane gallium nitride was induced in the 3D-structure by epitaxial lateral overgrowth, which enabled successful performance of our work. This result allowed us to axially decompose the strain in c-plane gallium nitride through Raman spectroscopy and establish a measurement technique for axially resolving the strain. This measurement technique is feasible using a conventional Raman spectrometer. Furthermore, the method was indicated to be applicable to all wurtzite-type crystals, including gallium nitride, silicon carbide, and aluminum nitride. Our work provides a new perspective for understanding the complex strain state in microareas for wurtzite materials. Comprehending the strain state, which strongly affects device performance, will help promote the research and development of III-V semiconductor devices.

Functional evaluation of BRCA1/2 variants of unknown significance with homologous recombination assay and integrative in silico prediction model.

Numerous variants of unknown significance (VUSs) exist in hereditary breast and ovarian cancers. Although multiple methods have been developed to assess the significance of BRCA1/2 variants, functional discrepancies among these approaches remain. Therefore, a comprehensive functional evaluation system for these variants should be established. We performed conventional homologous recombination (HR) assays for 50 BRCA1 and 108 BRCA2 VUSs and complementarily predicted VUSs using a statistical logistic regression prediction model that integrated six in silico functional prediction tools. BRCA1/2 VUSs were classified according to the results of the integrative in vitro and in silico analyses. Using HR assays, we identified 10 BRCA1 and 4 BRCA2 VUSs as low-functional pathogenic variants. For in silico prediction, the statistical prediction model showed high accuracy for both BRCA1 and BRCA2 compared with each in silico prediction tool individually and predicted nine BRCA1 and seven BRCA2 variants to be pathogenic. Integrative functional evaluation in this study and the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG/AMP) guidelines strongly suggested that seven BRCA1 variants (p.Glu272Gly, p.Lys1095Glu, p.Val1653Leu, p.Thr1681Pro, p.Phe1761Val, p.Thr1773Ile, and p.Gly1803Ser) and four BRCA2 variants (p.Trp31Gly, p.Ser2616Phe, p.Tyr2660Cys, and p.Leu2792Arg) were pathogenic. This study demonstrates that integrative evaluation using conventional HR assays and optimized in silico prediction comprehensively classified the significance of BRCA VUSs for future clinical applications.

Total Synthesis of Violaceoid A and (-)- and (+)-Violaceoid B.

The first total synthesis of violaceoid A, a cytotoxic agent, and the asymmetric total synthesis of (-)- and (+)-violaceoid B are reported. The precursor was accessed by desymmetrization of a substituted quinol moiety, and the racemic secondary alcohol was kinetically resolved using a chiral nucleophilic catalyst. The asymmetric synthesis of (-)- and (+)-violaceoid B elucidated the absolute configuration of the naturally occurring violaceoid B. Synthetic violaceoid A inhibited the growth of human breast cancer cell lines MCF-7 and Hs 578T at concentrations of less than 100 µM, while ( S)- and ( R)-violaceoid B were inactive.

Pt thermal atomic layer deposition for silicon x-ray micropore optics.

We fabricated a silicon micropore optic using deep reactive ion etching and coated by Pt with atomic layer deposition (ALD). We confirmed that a metal/metal oxide bilayer of Al2O3∼10 nm and Pt ∼20 nm was successfully deposited on the micropores whose width and depth are 20 µm and 300 µm, respectively. An increase of surface roughness of sidewalls of the micropores was observed with a transmission electron microscope and an atomic force microscope. X-ray reflectivity with an Al Kα line at 1.49 keV before and after the deposition was measured and compared to ray-tracing simulations. The surface roughness of the sidewalls was estimated to increase from 1.6±0.2 nm rms to 2.2±0.2 nm rms. This result is consistent with the microscope measurements. Post annealing of the Pt-coated optic at 1000°C for 2 h showed a sign of reduced surface roughness and better angular resolution. To reduce the surface roughness, possible methods such as the annealing after deposition and a plasma-enhanced ALD are discussed.

Polarization-independent high-index contrast grating and its fabrication tolerances.

A polarization-independent, high-index contrast grating (HCG) with a single layer of cross stripes allowing simple fabrication is proposed. Since the cross stripes structure can be suspended in air by selectively wet-etching the layer below, all the layers can be grown at once when implemented for vertical-cavity surface-emitting lasers. We optimized the structure to have a broad and high reflectivity band centered at around 1 µm using a finite difference time domain method, and obtained an 80 nm high reflectivity band centered at 0.97 µm in which the reflectivity exceeded 99.5%. We also investigated the fabrication tolerances of the structure and found that, assuming careful optimizations of electron beam lithography for the precise grating width and dry-etching for the vertical sidewall, the suggested polarization-independent HCG can be fabricated using standard technologies.

Effects of gender and aging in patients who undergo coronary artery bypass grafting: from the FU-Registry.

BACKGROUND: It is unclear whether gender and aging influence the characteristics of patients who undergo coronary artery bypass grafting (CABG). METHODS: We retrospectively reviewed a clinical database of 1,498 patients (male/female = 1133/365, age 67 ± 9 years) who underwent CABG at Fukuoka University Hospital from 1994 to 2010. RESULTS: Male showed significantly younger, higher percentages (%) of smoking and hyperuricemia (HU), higher levels of serum creatinine, and lower % hypertension (HT) and diabetes mellitus (DM), and lower levels of left ventricular ejection fraction than female. In multivariate analysis, all parameters identified independent variables associated with the gender difference. Next, we divided the patients into 5 groups according to age, and each group was then separated by gender. The % of males significantly decreased with aging, whereas % female significantly increased. Although % smoking and estimated glomerular filtration rate (eGFR), and body mass index (BMI) in all patients, males and females significantly decreased with aging, HU, left ventricular end diastolic pressure and the number of significantly stenosed coronary vessels were not associated with gender or aging. Interestingly, % HT in all patients and males significantly increased with aging, whereas that in females was not associated with aging. Serum low-density lipoprotein cholesterol levels in males significantly decreased with aging, while those in all patients and females were not associated with aging. In this contemporary data set, the decreases in % smoking and eGFR with aging were common characteristics in male and female patients. In addition, there were gender and aging differences in % smoking, % HT, BMI and eGFR, whereas no differences were observed in % DM, % dyslipidemia or % HU. CONCLUSIONS: Before CABG, high-risk patients with coronary artery disease who is going to undergo CABG may need to be managed more strictly considering to gender and age to avoid CABG.

Efficacy of propafenone hydrochloride in preventing postoperative atrial fibrillation after coronary artery bypass grafting.

BACKGROUND: Atrial fibrillation (AF) is one of the most common complications after coronary artery bypass grafting (CABG), and the incidence of postoperative AF (PAF) is estimated to range from 10% to 40%. PAF is a serious complication that is related to unstable hemodynamics, development of embolisms, patient discomfort, and increased medical costs associated with the prolongation of hospital stay. Sometimes, immediate attention is also necessary. In this study, we assessed the efficacy of treatment with the antiarrhythmic drug propafenone hydrochloride, which was administered in the early postoperative period, in preventing the development of PAF, and we attempted to identify risk factors for PAF. MATERIALS AND METHODS: The subjects were 78 patients who underwent isolated off-pump CABG between July 2007 and October 2008. We conducted the study by dividing the patients into 2 groups, a group of 26 patients who received propafenone hydrochloride (P group) and a control group of 52 patients who did not receive this drug (C group). The patients in the P group were given propafenone hydrochloride (150-450 mg/day orally) for 10 days, starting on the day after surgery, and were observed for the development of AF by means of continuous 12-lead electrocardiographic monitoring. Development of AF was defined as AF that lasted

Endoscopic radial artery harvesting for coronary artery bypass grafting: the initial clinical experience and results of the first 50 patients.

BACKGROUND: The radial artery (RA) is a commonly used arterial conduit in coronary artery bypass grafting (CABG). Traditional open-vessel harvest often leads to postoperative wound complications and cosmetic problems. Endoscopic RA harvesting (ERAH) has been widely used to prevent these problems. The purpose of this study was to assess these problems and graft patency in the first 50 patients who underwent ERAH. METHODS: Between February 2006 and October 2007, 50 patients underwent ERAH with the VasoView system (Boston Scientific). These patients were compared with 50 patients who underwent the traditional open technique. RESULTS: The mean age was 62.8 years in both groups. All RAs were successfully harvested. No conversion was made from ERAH to the traditional open technique. The mean harvesting time (forearm ischemic time) was 27.4 + or - 6.5 minutes, and the mean length of the RA in the ERAH group was 18.5 cm. Neither wound complications, such as wound infection and skin necrosis, nor severe neurologic complications were recorded. The patency rate was 95.9% (95/99) in the ERAH group and 94% (94/100) in the open group. CONCLUSION: ERAH can be performed safely, and the early results are satisfactory. Endoscopic vessel harvesting is therefore recommended as the technique of choice for RA harvesting.

New method of thermal coronary angiography for intraoperative patency control in off-pump and on-pump coronary artery bypass grafting.

BACKGROUND: We evaluated the effectiveness of a new thermal coronary angiogram system using intraoperative imaging with an infrared camera for coronary artery bypass grafting. METHODS: The thermal coronary angiograms of 51 patients who underwent a total of 107 coronary artery bypass grafts were evaluated. Thermal coronary angiograms were obtained after completing distal anastomoses by the injection of cold saline solution into the vein grafts or free arterial grafts or by reperfusion with warmer blood in the internal thoracic artery grafts. Temperature differences of greater than 0.1 degrees C between the injectant and the epimyocardium resulted in high-contrast images. RESULTS: Thermal coronary angiograms were obtained from 107 coronary artery bypass grafts; 103 grafts were patent (96.3%), and 2 internal thoracic artery grafts were occluded. After reanastomoses, thermal coronary angiograms were again obtained, and all grafts appeared to be patent. Four grafts did not clearly show hemokinesis because of an intramyocardial segment or circumferential fat surrounding the artery. CONCLUSIONS: Thermal coronary angiograms cannot show hemokinesis clearly in cases with an intramyocardial arterial segment or in patients in whom the grafts are surrounded by fat. Therefore, thermal coronary angiograms are considered to play a valuable role in confirming the success or failure of myocardium revascularization because this diagnostic modality does not interfere with the surgical procedures, is noninvasive, and can be both quickly and easily performed.

Coronary artery bypass grafting in DES era.

Coronary artery bypass grafting (CABG) has played an important role in the treatment of ischemic heart disease. Recently, the introduction of a drug-eluting stent (DES) has decreased the incidence of restenosis after percutaneous intervention (PCI). PCI with a DES is being increasingly performed, whereas the number of patients for whom CABG has been indicated has decreased over the last few years in Japan and the United States. According to a report, the number of patients undergoing CABG will not decrease in the future due to its usefulness in the treatment of multi-vessel lesions. We have also reviewed how CABG should be improved. For this purpose, it may be important to carry out less invasive CABG by the off-pump method and to improve the long-term results obtained by CABG with an internal thoracic artery graft and complete revascularization. Hence, CABG may achieve better long-term results compared with PCI and continued future application of CABG.

[Posterior ventricular septal perforation successfully repaired through right ventricular approach].

A 65-year-old man underwent a successful repair of a posterior ventricular septal perforation (VSP) 9 days after suffering an acute inferior myocardial infarction. After hospitalization, his hemodynamic condition gradually worsened, in spite of administering intensive medical therapy. Emergent operation was performed on the 4th day after onset. An equine pericardial patch was sutured around the VSP through the right ventricular side of the septum using the double-patch repair method and the right ventricular wall was closed as using the standard extracorporeal perfusion technique. The dimensions of the VSP measured 5 mm in diameter. Transesophageal echocardiography was performed on the 14th postoperative day. Cardiac catheter examination was done on the 18th postoperative day. No residual shunt was recognized and cardiac function was good. He was discharged on the 20th postoperative day. The occurrence of a posterior VSP is comparatively rare, and repair of VSP is difficult to perform during an acute period. Therefore, the operative results of VSP cases remain poor.

Application of a new suture material called the U-Clip for composite and sequential grafting with off-pump coronary bypass surgery.

BACKGROUND: The U-Clip was found to facilitate the interrupted anastomosis of coronary artery bypass grafts (CABG). This device may be beneficial especially in multivessel off-pump CABG (OPCAB) using composite grafts or sequential anastomosis. The aim of this study was to evaluate our early clinical experience using the U-Clip in OPCAB cases. METHODS: This retrospective study included 118 patients who underwent off-pump CABG between 2001 and 2004. The mean age of the 91 men and 27 women was 69.5 +/- 8.0 years (range, 47-85). The U-Clip was adopted for sewing 73 proximal ends of the free graft to the side or end of the inflow conduit to prepare the composite graft. The U-Clip was also applied to 112 distal anastomoses, especially to the side-to-side anastomosis of the sequential graft. RESULTS: Hospital mortality rate was 0.8% (1/118). The early patency rate of distal anastomoses using the U-Clip was 95% (96/101). The early patency rate of proximal anastomoses using the U-Clip was 98.4% (62/63). Interim angiography was performed in 12 patients (range, 3.8-42 months; average, 16 months). In these 12 patients, a total of 8 proximal anastomoses of conduits using the U-Clip were all patent without stenosis. The patency rate of a total of 10 distal anastomoses in the 12 patients using the U-Clip was 100%. CONCLUSIONS: The U-Clip-interrupted anastomosis enables a safe, definite, and rapid end-to-end or end-to-side connection of arterial grafts. We therefore consider the U-Clip to be a useful suture material especially for multivessel OPCAB using multiple arterial grafts.

ApoA-I/phosphatidylcholine discs remodels fast-migrating HDL into slow-migrating HDL as characterized by capillary isotachophoresis.

OBJECTIVE: Capillary isotachophoresis (cITP) is a technique for characterizing plasma lipoprotein subfractions according to their electrophoretic charges. We used this technique to examine the mechanism by which apoA-I/phosphatidylcholine (POPC) discs increase pre-beta HDL. METHODS AND RESULTS: The cITP analysis was performed using plasma prestained with a lipophilic dye on a Beckman P/ACE MDQ system. Plasma from a patient with lecithin:cholesterol acyltransferase (LCAT) deficiency who had increased apoE-containing HDL was used to characterize the charge distribution of apoA-I/POPC discs. cITP analysis of apoB- and E-depleted plasma of the patient in the presence of apoA-I/POPC discs indicated two major subfractions of apoA-I/POPC discs with mobilities of triglyceride-rich lipoproteins (fast and slow apoA-I). Incubation of whole plasma from a normolipidemic subject in the presence of apoA-I/POPC discs caused a reduction in cITP fast (f)- and intermediate (i)-migrating HDL, and fast and slow apoA-I, and an increase in slow (s)-migrating HDL. The changes in cITP lipoprotein subfractions were not affected by the inhibition of LCAT activity. ApoA-I/POPC discs increased the fractional esterification rate of cholesterol in apoB-depleted plasma. CONCLUSION: ApoA-I/POPC discs remodeled cITP fHDL and iHDL to sHDL independent of LCAT activity.

Central arterial cannulation and the arch first method for aortic arch aneurysm repair.

BACKGROUND: We investigated whether the axillary artery or ascending aorta cannulation combined with the arch first method decreases the risk of stroke during total arch replacement. PATIENTS AND METHODS: From January 2002 to January 2006, 35 total arch replacements were performed with the arch first method and central arterial cannulation. The mean age was 66+/-10 years. The cannulation sites were the axillary artery in 19 and the ascending aorta in 16. The arch first method (a short period of deep hypothermic circulatory arrest with retrograde cerebral perfusion and then subsequent antegrade cerebral perfusion) was used in all patients. RESULTS: The mean retrograde cerebral perfusion time was 29+/-7 min. The incidence of the permanent neurological dysfunction related to the surgical procedures was 2.9% (1/35). Hospital mortality was 5.7% of patients (2/35). There was no difference in the operative outcome between the 2 arterial inflow sites. CONCLUSION: At the time of total arch replacement, the use of central arterial cannulation and the arch first method are effective methods for preventing permanent brain injury. Cannulation of the ascending aortic using Dispersion cannula perfusing toward the aortic valve is considered to be a safe and favorable method for central arterial cannulation.

Higher small arterial elasticity in hypertensive patients treated with angiotensin II receptor blockers.

Although evidence from basic research suggests the involvement of angiotensin II (Ang II) in the progression of arteriosclerosis, the clinical data are limited. In the present study, hypertensive outpatients who were well controlled with antihypertensive medication and had similar blood pressure levels were studied, and arterial elasticity was compared between those receiving Ang II receptor blockers (ARBs) and those treated with other antihypertensive agents. The effects of HMG-CoA reductase inhibitors (STs) on arterial elasticity were also evaluated. The study enrolled 298 outpatients who had been diagnosed with essential hypertension whose blood pressure was controlled to 150/95 or less by antihypertensive treatment (excluding angiotensin converting enzyme [ACE] inhibitors) for at least 2 months. The small artery elasticity index (SAEI) was determined for each patient from the radial artery pulse waves using a non-invasive pulse wave analysis system CR-2000. The mean of two blood pressure measurements taken from subjects lying in a recumbent position during SAEI analysis was used for the data analysis. The patients were grouped according to the use of ARBs and STs, and two-way analysis of variance (ANOVA) was used for statistical comparisons. A backward stepwise multiple regression analysis was carried out to identify factors contributing to the SAEI. Hypertensive patients receiving ARB treatment had a significantly higher SAEI compared to those not receiving ARBs, despite the similar blood pressure levels of both groups. No significant effects of ST treatment on the SAEI were observed (two-way ANOVA). A backward stepwise multiple regression analysis for the SAEI suggested that ARB treatment was an independent determinant of the SAEI after the adjusting of age, gender, total cholesterol, high density lipoprotein cholesterol, smoking and systolic blood pressure. Our results suggested that while providing blood pressure control similar to that of other antihypertensive agents, ARBs may also increase vascular elasticity and thereby delay the progression of arteriosclerosis.

Influence of age and hypertension on the association between small artery compliance and coronary artery disease.

Small artery elasticity index (SAEI) as determined by a new non-invasive pulse-wave contour analysis has been used to identify abnormalities of the arterial wall associated with aging, hypertension (HT), endothelial dysfunction, and coronary artery disease (CAD). The present study examined the influence and interaction of CAD risk factors on the association between SAEI in risk-associated patients with CAD (case subjects, n = 178) and without CAD (control subjects, n = 202). Case subjects had a lower SAEI than control subjects, and age was consistently and negatively correlated with SAEI. The HT was related to reduced SAEI in female subjects overall and in male case subjects. In male patients with hypertension, the association between SAEI and CAD was significant (P < .05) after considering conventional risk factors of CAD. In conclusion, age and HT should be considered when using SAEI in the early diagnosis of CAD, and lower SAEI could be of greatest diagnostic value in male patients with HT.

Inhibition of cholesteryl ester transfer protein activity by JTT-705 increases apolipoprotein E-containing high-density lipoprotein and favorably affects the function and enzyme composition of high-density lipoprotein in rabbits.

BACKGROUND: Inhibition of cholesteryl ester transfer protein (CETP) is an efficient way to increase high-density lipoprotein (HDL) levels in humans. We investigated the effects of the inhibition of CETP activity by a CETP inhibitor, JTT-705, on the function and composition of HDL particles. METHODS AND RESULTS: Japanese white rabbits were fed either normal rabbit chow LRC-4 (n=10) or a food admixture of LRC-4 and 0.75% JTT-705 (n=10) for 7 months. JTT-705 significantly inhibited CETP activities, increased HDL cholesterol (HDL-C) levels and the ratio of HDL2-C/HDL3-C, and decreased the fractional esterification rate of cholesterol in HDL, indicating preferentially increased large HDL particles. Treatment with JTT-705 increased all of the 3 charge-based HDL subfractions as determined by capillary isotachophoresis: fast-migrating, intermediate-migrating, and slow-migrating HDL. The percentage of slow HDL, ie, apolipoprotein E (apoE)-containing HDL and levels of apoE in HDL fraction, was also increased. JTT-705 treatment increased serum paraoxonase activity and HDL-associated platelet-activating factor acetylhydrolase activity, but decreased the plasma lysophosphatidylcholine concentration. CONCLUSIONS: Inhibition of CETP activity by JTT-705 not only increased the quantity of HDL, including HDL-C levels and charge-based HDL subfractions, but also favorably affected the size distribution of HDL subpopulations and the apolipoprotein and enzyme composition of HDL in rabbits.

Calcification of the medial layer of the internal thoracic artery in diabetic patients: relevance of glycoxidation.

OBJECTIVE: The aim of this study was to evaluate the role of glycoxidation in the calcification of the internal thoracic artery (ITA) in diabetes mellitus (DM). METHODS: ITA samples were obtained from 17 patients with type 2 DM (age 62.9 +/- 10.5 years) and 12 age-matched, nondiabetic patients (age 62.5 +/- 10.2 years) who underwent coronary artery bypass grafting. These samples were analyzed histopathologically and assessed for calcification by von Kossa staining and for glycoxidation by immunohistochemistry using anti-N(epsilon)-(carboxymethyl)lysine (CML) antibody. Morphometric evaluation of calcification of the medial layer, intimal thickness and intima-to-media ratio was performed using NIH image software. To evaluate the mechanism of the interaction between calcification and glycoxidation, we developed an in vitro model of calcification of collagen that was chemically modified by glucose, glutaraldehyde or epoxy compound. The calcium-binding activity of these collagens was determined in hydrolysates using atomic absorption spectrophotometry. RESULTS: ITAs of both diabetic and nondiabetic patients were free of atherosclerosis, and no differences were found between the two groups with regard to intimal thickness and intima-to-media ratio. Areas of calcification were noticed in both groups in the tunica media, but not in the tunica intima. Calcium deposits were localized within the extracellular matrix, which was immunohistochemically positive for CML. The extent of medial layer calcification was significantly greater in diabetic patients than nondiabetic subjects, but was independent of known risk factors such as hypertension, hyperlipidemia, obesity and history of old myocardial infarction. The binding activity of collagen was time-dependently increased with in vitro incubation of glucose. A significant increase in the calcium-binding ability was observed in glucose- and glutaraldehyde-modified collagens, but not in epoxy compound-modified collagen. CONCLUSION: Our results suggest that glycoxidative modification of the extracellular matrix, in particular collagen, of the vascular wall may enhance the development of ITA calcification in diabetic patients.

Commissural dehiscence and pannus formation of porcine heart valve bioprostheses.

From May 1976 to June 2001, 95 porcine heart valve bioprostheses (BPs) were explanted. Among them, 85 prostheses were available for an analysis of structural valve deterioration (SVD). There were 32 Hancock (H) BPs and 53 Carpentier-Edwards (CE) BPs, with a mean implantation duration of 12 +/- 3.5 and 11.0 +/- 4 years, respectively. There were two extraordinary complications. One was commissural dehiscence (CD) of the CEBPs in 10/53 cases (18.9%), with a mean duration of 12.7 +/- 2.7 years. This was not observed in the HBPs. The other was pannus formation in both BPs. In the mitral position, it was 25/32 (78.1%) in HBPs and 37/53 (69.8%) in CEBPs. In the aortic position, it was 4/32 (1.3%) in the HBPs and 11/53 (20.8%) in the CEBPs. We conclude that CD could not be diagnosed preoperatively and anticoagulation is therefore indicated to prevent pannus formation of such prostheses, particularly in those located in the mitral position.