Collagen vitrigel promotes hepatocytic differentiation of induced pluripotent stem cells into functional hepatocyte-like cells.

Differentiation of stem cells to hepatocytes provides an unlimited supply of human hepatocytes and therefore has been vigorously studied. However, to date, the stem cell-derived hepatocytes were suggested to be of immature features. To obtain matured hepatocytes from stem cells, we tested the effect of culturing human-induced pluripotent stem (hiPS) cell-derived endoderm cells on collagen vitrigel membrane and compared with our previous reported nanofiber matrix. We cultured hiPS cell-derived endoderm cells on a collagen vitrigel membrane and examined the expression profiles, and tested the activity of metabolic enzymes. Gene expression profile analysis of hepatocytic differentiation markers revealed that upon culture on collagen vitrigel membrane, immature markers of AFP decreased, with a concomitant increase in the expression of mature hepatocyte transcription factors and mature hepatocyte markers such as ALB, ASGR1 Mature markers involved in liver functions, such as transporters, cytochrome P450 enzymes and phase II metabolic enzymes were also upregulated. We observed the upregulation of the liver markers for at least 2 weeks. Gene array profiling analysis revealed that hiPS cell-derived hepatocyte-like cells (hiPS-hep) resemble those of the primary hepatocytes. Functions of the CYP enzyme activities were tested in multi-institution and all revealed high CYP1A, CYP2C19, CYP2D6, CYP3A activity, which could be maintained for at least 2 weeks in culture. Taken together, the present approach identified that collagen vitrigel membrane provides a suitable environment for the generation of hepatocytes from hiPS cells that resemble many characteristics of primary human hepatocytes.

Factors affecting the incidence and outcome of Trueperella pyogenes mastitis in cows.

The main factors affecting the outcome of Trueperella pyogenes (T. pyogenes) mastitis were examined through a survey of diagnostic data and interviews relating to the occurrence of T. pyogenes mastitis in 83 quarters from 82 Holstein cows between August 2012 and April 2014. Ultimately, one cow was sold during the examination, and 82 quarters from 81 cows were used for analysis on prognosis. T. pyogenes mastitis occurred year round in both lactating and dry cows. The incidence of T. pyogenes mastitis did not significantly differ by month or show seasonality in either lactating or dry cows. Therefore, the occurrence of T. pyogenes mastitis also differed from that of summer mastitis. The 1-month survival rate of infected cows was 64.6% (53/82), and the recovery rate of quarters with T. pyogenes mastitis was 14.6% (12/82). Bivariate logistic regression analysis was performed with survival and culling of infected cows as objective variables and with recovery and non-recovery of quarters with T. pyogenes mastitis as objective variables. The severe cases were significantly culled (odds ratio, 16.30) compared to mild cases, and the status of quarters didn't recover (odds ratio, 6.50). The results suggest that mild to moderate symptom severity at the time of onset are the main factors affecting outcomes in cows and recovery of quarters infected with T. pyogenes mastitis. Further, high level of NAGase activity also suggested the potential use as an indicator of culling of cows with T. pyogenes mastitis.

[New Intracoronary Shunt Cannula in Off-pump Coronary Artery Bypass Grafting].

In off-pump coronary artery bypass grafting(OPCAB), a bloodless operative field has great influence on the quality of anastomosis. In addition, maintenance of distal coronary flow during anastomosis stabilizes hemodynamics. We introduced a new intracoronary shunt cannula, Mini Shunt Pro (MSP). MSP adopts the step-tip, which allows easy insertion and reduces the risk of intimal injury. The distal and proximal sides of the tips of MSP are 0.25 mm different in diameter, which improves its fitness to the coronary artery and enhances a bloodless operative field. This new shunt cannula is considered to be safe and useful for high-quality anastomosis in OPCAB.

Histomorphologic superiority of internal thoracic arteries over right gastroepiploic arteries for coronary bypass.

OBJECTIVE: In this study, we compared the histologic and morphometric properties of both internal thoracic arteries and the right gastroepiploic artery (GEA) in patients undergoing coronary artery bypass grafting (CABG). METHODS: We microscopically examined transverse sections of segments of both internal thoracic arteries and the right GEA obtained from 83 consecutive patients who underwent CABG. RESULTS: There were no significant differences between the internal thoracic arteries. Significant differences were found between the left and right internal thoracic arteries and GEA in the intimal width (21.8, 21.5, and 71.7 µm, respectively; P < .01), intima-to-media ratio (0.286, 0.256, and 0.749, respectively; P < .01), and media width (148.5, 157.5, and 164.8 µm, respectively; P = .43). No atherosclerotic lesions, medial calcification, or intimal thickening were seen in the internal thoracic arteries; however, atherosclerotic lesions were seen in the GEA. The intima of the GEA was thicker than that of the internal thoracic arteries. Intimal thickening of the GEA, but not the internal thoracic arteries, was positively correlated with risk of arteriosclerosis. In patients with diabetes mellitus, dietary/drug therapy and insulin therapy were associated with GEA intimal thickness (P = .02 and .01, respectively). CONCLUSIONS: The internal thoracic arteries have equivalent histologic and morphometric properties that differ from those of the GEA only in intimal width. The former had no intimal thickening, and is thus preferable to the GEA for CABG.

Utility of three-dimensional transesophageal echocardiography for diagnosis of unroofed coronary sinus.

We report a case of unroofed coronary sinus (URCS) in a 42-year-old female. At an outpatient clinic, she was found to have an atrioventricular septal defect and mitral regurgitation with pulmonary hypertension, and she was transferred to our institute for surgical treatment. Both atrioventricular valves were located at the same level, and both the right atrium (RA) and right ventricle were enlarged on two-dimensional transthoracic echocardiography. Color Doppler imaging demonstrated severe mitral and tricuspid regurgitation and a left-to-right shunt from the left atrium (LA) to the RA. Although an ostium primum defect of the atrial septum was suspected, the exact position of the shunt flow was unclear. Two-dimensional transesophageal echocardiography (2D-TEE) could visualize a direct communication between the LA and coronary sinus. Three-dimensional transesophageal echocardiography (3D-TEE) clearly visualized the entire route from the coronary sinus into the LA and RA. The utility of 3D-TEE as a modality complementary to 2D-TEE in diagnosis of URCS was confirmed.

[Mitral Valvuloplasty of Infective Endocarditis During Pregnancy].

This report describes a case in which we treated a patient who developed infective endocarditis in the mitral valve at 28 weeks' gestation. The condition was resolved by performing mitral valvuloplasty 2 days after an emergency cesarean section. Although the patient was in a relatively stable period at 32 weeks' gestation, the mother had an extremely high risk of embolism;thus, emergency surgery was required. We believe that an accurate diagnosis in a timely manner and a valvuloplasty shortly after cesarean section saved the life of the mother and child.

Stress echocardiographic assessment of mitral valve function repaired using rough-zone trimming.

BACKGROUND: We invented novel mitral valve repair technique; rough-zone trimming procedure (RZT) for anterior mitral valve prolapse. Prolapse site was resected in obtuse triangle shape and sutured edges to creates deep coaptation and improves regurgitation. Though it is simple and reproducible technique, functional mitral stenosis is a risk. Valve function and hemodynamics were investigated using dobutamine stress echocardiography (DSE) in patients after mitral valve repair using RZT. METHODS: Patients underwent RZT for the anterior mitral valve (AML, n = 10), quadrangular resection (QR) of the posterior mitral valve (PML; n = 4), RZT + QR of bileaflet valves (bileaflet; n = 4) and healthy individuals (control; n = 10) and were assessed by DSE (doses up to 20 µg/Kg/min). Echocardiographic data including mitral valve area (MVA), mitral valve mean pressure gradient (MVmeanPG), and systolic pulmonary artery pressure (sPAP) were measured at rest and at peak stress. RESULTS: Rest/stress MVA (cm(2)), MVmeanPG (mmHg) and sPAP (mmHg) were 2.8 ± 0.4 and 3.4 ± 0.3, 3.3 ± 1.1 and 7.4 ± 4.1, and 25.7 ± 4.7and 49.1 ± 4.1, respectively, in the AML group. Dobutamine stress increased all parameters but not to pathological levels. The results were similar to those of the other groups after mitral valve repair, whereas MVA was larger and MVmeanPG was lower in the control than in the AML group. CONCLUSIONS: Valve repair using RZT does not pathologically obstruct the mitral valve, either at baseline or during dobutamine stress, and does not affect valve hemodynamics and reserve.

New parameter of the right gastroepiploic arterial graft using the power spectral analysis device named MemCalc soft.

OBJECTIVES: Transit-time flow measurement (TTFM) parameters such as mean graft flow (MGF, ml/min), pulsatility index (PI) and diastolic filling (DF, %) have been extensively researched for internal mammary arterial or saphenous vein grafts. In our experience of using the right gastroepiploic arterial (GEA) graft for right coronary artery (RCA) grafting, we observed unique GEA graft flow waveforms. We analysed the GEA graft flow waveforms for their effectiveness in determining GEA graft patency by power spectral analysis. METHODS: Forty-five patients underwent off-pump coronary artery bypass using the GEA graft for RCA grafting individually. The means of intraoperative MGF, PI and DF were compared between patent and non-patent grafts, postoperatively. Furthermore, the GEA flow data were output and analysed using power spectral analysis. RESULTS: Forty grafts were 'patent' and five were 'non-patent'. There were no significant differences in the mean TTFM parameters between the patent and non-patent grafts (MGF: 22 vs 8 ml/min, respectively, P = 0.068; PI: 3.5 vs 6.5, respectively, P = 0.155; DF: 63 vs 53%, respectively, P = 0.237). Results of the power spectral analysis presented clear differences; the power spectral density (PSD) of patent grafts presented high peaks at frequency levels of 1, 2 and 3 Hz, and the non-patent graft PSD presented high peaks that were not limited to these frequencies. The PSD had a sensitivity and specificity of 80 and 87.5%, respectively. CONCLUSIONS: Power spectral analysis of the GEA graft flow is useful to distinguish between non-patent and patent grafts intraoperatively. This should be used as a fourth parameter along with MGF, PI and DF.

Impact of novel intraoperative evaluation for mitral valve regurgitation: the retrograde cardio-protective beating test.

PURPOSE: The saline injection test is commonly employed during mitral valvuloplasty. However, discrepancies in regurgitation between the naked eye findings during the saline injection test and the postoperative echocardiographic findings have been noted. Here, we describe the new retrograde cardio-protective beating test (RC-beating test) which allows direct transatrial evaluation of the valve in the fully loaded, beating heart under cross-clamping by means of the retrograde perfusion of warm oxygenated blood into the coronary sinus. METHODS: From January 2006 to June 2012, 63 patients (mean age, 59 ± 15 years) with degenerative mitral valve regurgitation underwent mitral valve repair with application of this novel evaluation. RESULTS: In 20 of the 63 patients (31.8%), minor residual leaks that were not remarkable in the saline injection test were detected during the RC-beating test and were completely corrected. Almost all patients (n = 61, 96.8%) revealed no mitral regurgitation on intraoperative transesophageal echocardiography. The results of the RC-beating test were satisfactory, and discrepancies were not recognized at all between this evaluation and the postoperative echocardiographic findings. The ultrastructure of the mitochondria showed significant myocardial preservation. CONCLUSION: The RC-beating test is a simpler and more accurate evaluation for mitral valve repair than the saline injection test. This novel evaluation enables a safe and more complete mitral valve repair.

Evaluation of gastroepiploic arterial grafts to right coronary artery using transit-time flow measurement.

OBJECTIVES: The objective of this study was to analyse the relationship between the intraoperative transit-time flow measurement (TTFM) parameter values and the postoperative angiographic results of gastroepiploic arterial (GEA) grafts to the right coronary artery (RCA). We investigated whether the intraoperative TTFM parameter values are reliable indicators of early patency in GEA grafts to the RCA. METHODS: Patients undergoing off-pump coronary artery bypass surgery with GEA grafts were included in this study. Eighty-three GEA grafts were individually anastomosed and examined by angiography 1 week after surgery. The quality of each graft was graded using FitzGibbon grading (Study 1) and graft-flow grading (Study 2). RESULTS: Study 1: Seventy-two grafts were determined as Grade A and 11 as Grades B or O. There were no significant differences in the average of mean graft flow (MGF), pulsatility index or diastolic filling percentage between Grade A and Grades B or O grafts. Study 2: Sixty-two grafts were graded as good-graft dominant, 16 as bidirectional and 5 as occlusion including string. The average of the MGF, pulsatility index and diastolic filling percentage in the grafts graded as bidirectional and occlusion including string were not significantly different from those of grafts graded as good-graft dominant. CONCLUSIONS: Previously reported cut-off values for intraoperative TTFM parameters could not be adapted for the early patency of GEA grafts to the RCA. However, the smoothness of the graft-flow curve may be a reliable predictor of postoperative graft patency.

[Intraoperative evaluation for residual mitral valve regurgitation; usefulness of the retrograde cardioprotective beating test].

Intraoperative evaluation is important for successful mitral valve plasty (MVP). We performed a saline injection test and a retrograde cardioprotective beating test (RC-beating test) for intraoperative evaluation. The concept of the RC- beating test is evaluation of residual mitral valve regurgitation( MR) under cardiac beating. A 66-year-old man with severe MR underwent MVP. The P3 chorda was ruptured and we performed quadrangular resection. The saline injection test showed trivial regurgitation. We then performed the RC-beating test and it revealed severe leakage from the posterior commissure(PC). Since the PC had a sclerotic change, another quadrangular resection was performed. Moreover,as the anterior leaflet( A3) was slightly elongated, the region was resected in an obtuse-angled triangle shape and repaired by suturing the edges. The final RC-beating test showed no residual leakage. The RC-beating test is useful for detecting residual mitral valve leakage.

Acute dilatation of the ascending aorta and aortic valve regurgitation in Loeys-Dietz syndrome.

Loeys-Dietz syndrome (LDS) is a recently recognized connective tissue disorder caused by mutations of the transforming growth factor (TGF)-ß receptors. It is an autosomal dominant syndrome characterized by the triad of arterial tortuosity and aneurysms, hypertelorism, and bifid uvula or cleft palate. We treated an 18-year-old woman with a 100-mm-diameter aortic root aneurysm and severe aortic valve regurgitation. She underwent urgent aortic root replacement and bioprosthetic valve implantation. LDS was diagnosed by postoperative genetic screening results. Histopathologic examination of the aortic wall showed diffuse degeneration and elastin fragmentation in the media.

Design and synthesis of a novel fluorescent protein probe for easy and rapid electrophoretic gel staining by using a commonly available UV-based fluorescent imaging system.

A new fluorescent molecular probe, methyl 3-(3,5-bis((bis(pyridin-2-ylmethyl)amino)-methyl)-4-hydroxyphenyl)-2-(5-(dimethylamino)naphthalene-1-sulfonamido) propanoate, dizinc(II) chloride salt (Dansyl-1-Zn(II)), which possesses Zn(II) complexes and a dansyl group, was designed and synthesized to enable the detection of proteins in solution and in high-throughput electrophoresis by using a UV-based detection system. Dansyl-1-Zn(II) exhibited weak fluorescence in the absence of proteins and strong green fluorescence at approximately 510 nm in the presence of BSA upon irradiation with light at a wavelength of 345 nm. Compared with conventional protocols for in-gel SDS-PAGE protein staining (e.g. silver staining, SYPRO Ruby, and Oriole), the operating times of which range from 90 min to overnight, Dansyl-1-Zn(II) allowed 1-step protein staining (SDS-PAGE →Staining →Detection) and shortened the operating time (35 min) with high sensitivity (LOD: 1 ng or less) under 312-nm or 365-nm light excitation with orange or red emission filters, respectively. Moreover, Dansyl-1-Zn(II) was successfully applied to protein identification by MS via in-gel tryptic digestion, Western blotting, and Native-PAGE. Accordingly, Dansyl-1-Zn(II) may facilitate highly sensitive and high-throughput protein detection, and it may be widely applicable as a convenient tool in various scientific and medical fields.

[Aortic valve surgery in a patient with cold agglutinin disease; effectiveness of continuous retrograde cardioplegia].

A 76-year-old female was hospitalized because of congestive heart failure and anemia. A thorough examination led to a diagnosis of severe aortic stenosis and cold agglutinin disease. The critical temperature for hemagglutination was 27 °C, which caused particular problems with regard to the myocardial protection temperature during surgery. Aortic valve replacement was performed safely by increasing the normal myocardial protection temperature from 15 °C to 32 °C and using 3 times the normal volume of cardioplagic fluid. As a result of strict perioperative thermal management, the operation was completed without any complications.

Mycotic aneurysm of ascending aorta in an infant.

Mycotic aneurysm of the ascending aorta is rare in infants. A 12-month-old infant developed a mycotic aneurysm of the ascending aorta secondary to group A streptococcus septicemia. He developed atrioventricular block, and the rapid aneurysm progression under medical therapy prompted us to perform an emergency operation. The aneurysm was successfully repaired using an autologous pericardial patch. He remained well with normal sinus rhythm and was developing normally at the 5-year follow-up.

Novel intraoperative evaluation for mitral valve regurgitation: retrograde cardioprotective beating test.

In mitral valvuloplasty, the saline injection test is commonly employed. However, discrepancies in regurgitation between the naked eye findings during the saline injection test and the postoperative echocardiographical findings are noted. Here, we describe a technique that allows direct transatrial evaluation of the valve in the fully loaded, beating heart without the risks of air embolism. Physiological systolic mitral valve movement is reproduced under aortic cross-clamping. This novel evaluation enables a complete and safe mitral valve repair.

[Bioprosthetic valve endocarditis associated with rapid progression of valvular stenosis;report of a case].

Long-term durability and stability of the hemodynamic performance of commercially available 2nd generation Carpentier-Edwards stented bovine pericardial aortic bioprostheses (Perimount valve) is reported based on echocardiographic follow-up examinations. Prosthetic valve deterioration of the Perimount valve usually occurs late and is age-dependent. We describe bioprosthetic valve deterioration of a Perimount valve with severe stenotic change 1 month postoperatively that developed in an infective endocarditis patient. Although preoperative echocardiography showed no signs of valve dehiscence or vegetation on the prosthesis, valve stenosis alone had progressed extremely rapidly during 2 weeks of febrile symptom. The valve leaflet was found at explant to be infiltrated with a cellular material fibrin other than bacteria or leukocytes. This rare occurrence needs to be reported in the literature to forewarn clinicians of an early manifestation of endocarditis that may not be recognized yet.

Surgical resection of primary cardiac lymphoma presenting as a huge mass: report of two cases.

Primary cardiac lymphoma (PCL) is a rare malignancy that is sometimes diagnosed at later stages because it is not associated with specific symptoms. Although chemotherapy is the standard for treating PCL, the value of surgical resection is controversial. We describe two patients who were treated by surgical resection and chemotherapy. Case 1 is a 37-year-old man with a history of acute lymphocytic leukemia and shortness of breath, and Case 2 is a 70-year-old woman with general fatigue. Both of them were presented at hospital. In Case 2, arrhythmic syncope occurred and direct current cardioversion was performed. Echocardiography showed a massive tumor in the right atrium and disturbed hemodynamics in both cases. The restricted venous return was alleviated by emergency surgery. The pathology report indicated primary cardiac lymphoma that was regressed by post-operative chemotherapy. A massive PCL should be surgically resected to prevent sudden death.

Repeat conduit replacement in the pulmonary position without sternal resplitting for the patient with repaired Tetralogy of Fallot and the absent inferior caval vein.

Recently, sternal reentry has been performed with low perioperative mortality and morbidity. In some patients, however, there are specific problems leading to life-threatening events during sternal reentry. A 27-year-old woman with repaired Tetralogy of Fallot and the absent inferior caval vein was referred to our department for pulmonary conduit replacement. Preoperative computed tomography disclosed the dominant right coronary artery and the ascending aorta longitudinally running just beneath the midline of the sternum. Therefore, we selected bilateral antero-lateral thoracotomy as the alternative approach to avoid the injury of these vital organs. The left axillar and right femoral artery were used for arterial perfusion sites. Venous drainage could be obtained from the left superior caval vein and the isolated hepatic vein via the left antero-lateral thoracotomy, and the right superior caval vein via the right antero-lateral thoracotomy. We successfully performed the repeat conduit replacement via the right antero-lateral thoracotomy.