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Objective To evaluate the efficacy of pleurodesis using OK-432 after cardiac surgery in the neonatal period or early infancy.Methods We retrospectively reviewed the data of 11 consecutive patients who underwent cardiac surgery in the neonatal period or early infancy and pleurodesis using OK-432 for persistent postoperative pleural effusion in two institutions.Results The median age at surgery was 8 days (interquartile range [IR], 2–18) with a body weight of 2.84 kg (IR, 2.30–3.07). The maximum amount of pleural drainage before pleurodesis was 94.7 (IR, 60.2–107.7) ml/kg/day. Pleurodesis was initiated at postoperative day 20 (IR, 17–22) and performed in bilateral pleural spaces in seven patients and unilateral in four. The median numbers of injection were 4 (IR, 3–6) times per patient and 3 (IR, 2–3) times per pleural space. In 10 patients, pleural effusion was decreased effectively, and drainage tubes were removed without reaccumulation within 15 (IR, 12–28) days after initial pleurodesis. However, in one patient, with severe lymphedema, pleural effusion was uncontrollable, resulting in death due to sepsis. Adverse events were observed in nine patients; temporal deterioration of lung compliance and arterial blood gas occurred in two, insufficient drainage requiring new chest tube(s) in five, temporal atrial tachyarrhythmia in one, and lymphedema in four.Conclusions Pleurodesis using OK-432 is effective and reliable for persistent postoperative pleural effusion in neonates and early infants. Most of the complications, which derived from inflammatory reactions, were temporary and controllable. However, severe lymphedema is difficult to control.

OVERVIEW: China has become one of the most desired locations in which to do R&D. However, it has little innovation of its own, and intellectual property protection is weak. This raises questions: Is China R&D more hype than reality? Do cost advantages really outweigh the risk of losing technology to Chinese competitors? Lessons learned from managing R&D in China show that in order to avoid the typical pitfalls of managing R&D in a developing country, any China-based R&D must be part of an overall China strategy and must also be part of a global R&D effort. KEYWORDS: International R&D, Emerging economies, China, China is not your typical developing country--or is it? Those who have recently traveled to Beijing, Shanghai or Shenzhen have seen cities with Western-level infrastructure, Manhattan-like skylines, city-wide public transportation [...]

A five-year-old boy was diagnosed with the ventricular septal rupture and ventricular aneurysm after blunt chest trauma in child abuse. Because of the intractable heart failure, he underwent operation in subacute period. Postoperative course was uneventful. The blunt cardiac injury in children can be caused by mild trauma and can be lethal. Surgical intervention should be considered when the clinical condition is unstable.

Various surgical techniques have been reported for Scimitar syndrome, because of the heterogenous anatomy of the disease. We developed a novel surgical method to repair Scimitar syndrome, in which, a new pulmonary venous route is constructed behind the inferior vena cava using autologous flaps of the inferior vena cava and the interatrial septum. An adult case of Scimitar syndrome was repaired by this method with good results.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.jtcvs.2010.02.006 Byline: Hiroyuki Muranaka, Akira Marui, Masaki Tsukashita, Jian Wang, Jota Nakano, Tadashi Ikeda, Ryuzo Sakata Abbreviations: AR, adrenergic receptor; DCM, dilated cardiomyopathy; DCM-UL, dilated cardiomyopathy (unloaded); DT, developed tension; LV, left ventricle (ventricular); LVAD, left ventricular assisted device; LVDd, left ventricular end-diastolic dimension; LVDs, left ventricular end-systolic dimension; mRNA, messenger RNA; TUNEL, terminal dUTP nick end labeling Abstract: Left ventricular assist devices are used in patients with end-stage dilated cardiomyopathy as a 'bridge to recovery.' However, physiologic and histologic changes under prolonged mechanical unloading have not been elucidated. Thus, we investigated these changes in the rat heart with dilated cardiomyopathy under mechanical unloading after heterotopic transplantation. Author Affiliation: Department of Cardiovascular Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan Article History: Received 3 September 2009; Revised 8 January 2010; Accepted 2 February 2010 Article Note: (footnote) Disclosures: None.

AbstractWe experienced a case of a 1-year-old female with congenital tricuspid valve regurgitation caused by tricuspid valve dysplasia. The anterior and septal leaflets were particularly dysplastic, and leaflet tethering was observed. The anterior papillary muscle was approximated to the interventricular septum, and a commissural edge-to-edge suture was inserted on the anteroseptal commissure. Tricuspid valve regurgitation improved to be trivial after surgery and has not worsened 1 year later. Papillary muscle approximation could be one of the feasible reparative techniques for congenital tricuspid valve regurgitation.

OVERVIEW: China has become one of the most desired locations in which to do R&D. However, it has little innovation of its own, and intellectual property protection is weak. This raises questions: Is China R&D more hype than reality? Do cost advantages really outweigh the risk of losing technology to Chinese competitors? Lessons learned from managing R&D in China show that in order to avoid the typical pitfalls of managing R&D in a developing country, any China-based R&D must be part of an overall China strategy and must also be part of a global R&D effort. KEY CONCEPTS: international R&D, emerging economies, management in China., China is not your typical developing country--or is it? Those who have recently traveled to Beijing, Shanghai or Shenzhen have seen cities with Western-level infrastructure, Manhattan-like skylines, city-wide public transportation [...]

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.jtcvs.2005.10.026 Byline: Akira Marui (a), Keiichi Hirose (a), Takayuki Maruyama (b), Yoshio Arai (a), Yuhong Huang (a), Kazuhiko Doi (c), Tadashi Ikeda (a), Masashi Komeda (a) Abbreviations: bFGF, basic fibroblast growth factor; BITA, bilateral internal thoracic artery; BMC, bone mineral content; BMD, bone mineral density; DEXA, dual-energy x-ray absorptiometry; PG, prostaglandin; PLGA, copoly lactic acid/glycolic acid; SWC, sternal wound complication Abstract: Sternal wound complications are devastating events occurring in coronary artery bypass surgery, particularly in patients with diabetes. Prostaglandin E.sub.2 receptors have 4 subtypes, and the activation of the EP4 receptor induces bone regeneration. The present study investigated the utility of a prostaglandin E.sub.2 EP4 receptor-selective agonist in sternal healing after median sternotomy with the removal of the bilateral internal thoracic arteries in diabetic rats. Author Affiliation: (a) Department of Cardiovascular Surgery, Kyoto University Graduate School of Medicine, Kyoto Japan (b) Minase Research Institute, Ono Pharmaceutical Company, Osaka, Japan (c) Takeda Hospital, Kyoto, Japan Article History: Received 28 May 2005; Revised 21 September 2005; Accepted 20 October 2005 Article Note: (footnote) This study is supported by a Grant for Scientific Research, from the Japanese Ministry of Education and Science.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.amjsurg.2004.08.067 Byline: Hiromitsu Jinno (a), Tadashi Ikeda (a), Sota Asaga (a), Takeshi Muto (a), Yuko Kitagawa (a), Hirofumi Fujii (b), Makio Mukai (c), Masaki Kitajima (a) Keywords: Sentinel lymph node; Breast cancer; Age; Particle size Abstract: Increasing age of breast cancer patients was reported to cause not only failure of sentinel lymph node (SLN) identification but also false negative results. Author Affiliation: (a) Department of Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160, Japan (b) Department of Radiology, Keio University School of Medicine, Tokyo, Japan (c) Department of Pathology, Keio University School of Medicine, Tokyo, Japan Article History: Received 11 March 2004; Revised 24 August 2004

This is a case report of a successful single‐lobe lung transplantation for pulmonary hypertension secondary to alveolar capillary dysplasia with misalignment of pulmonary veins (ACD/MPV). A 6‐year‐old boy underwent living‐donor single‐lobe transplantation with the right lower lobe from his 31‐year‐old mother. The pretransplantation graft size matching was acceptable: the estimated graft forced vital capacity (FVC) was 96.5% of the recipient's predicted FVC, and the graft size measured by computed tomography (CT) volumetry was 166% of the recipient's chest cavity volume. Right pneumonectomy followed by implantation was performed under cardiopulmonary bypass (CPB). The pulmonary arterial pressure was significantly decreased to 31/12 mm Hg immediately after transplantation, and the first PaO2/FiO2in the intensive‐care unit (ICU) was 422 mm Hg. Lung perfusion scintigraphy showed 97.5% perfusion to the right implanted lung 3 months after transplantation. Chest CT showed a mass rapidly growing in the native left upper lobe 6 months after transplantation, which was diagnosed as posttransplant lymphoproliferative disorder (PTLD) by a CT‐guided biopsy. After immunosuppressant reduction and six courses of chemotherapy with rituximab, he underwent native left upper lobectomy for salvage lung resection 13 months after transplantation. Seven months after lobectomy, he has returned to normal school life without any sign of tumor recurrence. Living‐donor single‐lobe lung transplantation can be a viable option for pulmonary hypertension secondary to alveolar capillary dysplasia with misalignment of pulmonary veins.

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.jtcvs.2007.03.021 Byline: Takeshi Shimamoto (a), Akira Marui (a), Kazuo Yamanaka (b), Norihiro Shikata (c), Keiichi Tambara (a), Tadashi Ikeda (a), Masashi Komeda (a) Author Affiliation: (a) Department of Cardiovascular Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan (b) Department of Cardiovascular Surgery, Koseikai Takeda Hospital, Kyoto, Japan (c) Department of Cardiology, Second Min-iren Central Hospital, Kyoto, Japan. Article History: Received 2 February 2007; Revised 5 March 2007; Accepted 15 March 2007

For patients with pulmonary atresia with ventricular septal defects and major aortopulmonary collateral arteries (PA/VSD/MAPCAs), unifocalization (UF) through median sternotomy during infancy has become a standard surgical treatment [1, 2].

ABO-incompatible (ABO-I) living-donor lobar lung transplantation (LDLLT) was successfully performed for a 14-year-old girl who suffered from bronchiolitis obliterans due to graft-versus-host disease following hematopoietic stem cell transplantation. In the ABO-I LDLLT procedure, the blood type O patient received a right lower lobe donated from her blood type B father and a left lower lobe donated from her blood type O mother. Desensitization therapy, using rituximab, immunosuppressants, and plasmapheresis, was implemented for 3 weeks prior to transplantation in order to reduce the production of anti-B antibodies in the recipient and prevent acute antibody-mediated rejection after ABO-I LDLLT.

Introduction:Autologous grafts for revascularization might be limited due to the quality of vessels. Decellularized xenogeneic vascular grafts potentially substitute for autologous grafts. We have been developing a decellularization method based on high hydrostatic pressure (HHP) which confers highly preserved extracellular structure. For broader application of the HHP-decellularized vascular graft, pre-implant endothelialization of the grafts by human endothelial cells would be anticipated which may prevent clot formation and graft occlusion.Hypothesis:We hypothesized to achieve sufficient ex vivoendothelialization of HHP-decellularized tissues by virtue of chemical endothelial surface modification and dynamic culture recapitulating blood flow.Methods:Thin slices of bovine aortic endothelium were decellularized by HHP. We divided the tissues in 2 groups: coated with E8 fragment of human laminin 411 (hL411) holding high affinity with endothelial cells and coated with gelatin (Gel). We directly seeded human umbilical vein endothelial cells (HUVECs) with a density of 2 х106cells/cm2, then divided the tissues in 2 subgroups: dynamic culture (20 cm/s) and static culture. We kept the culture for up to 2 weeks and evaluated the area coverage by staining of live cells by Calcein-AM, histological analysis, and scanning electron microscopy (SEM). We tested clot formation using human whole blood.Results:hL411/dynamic group exhibited significantly higher area coverage compared to those of other groups which almost reached 100% [coverage at 2 weeks: Gel/static vs hL411/static vs Gel/dynamic vs hL411/dynamic: 57.3±4.1 vs 61.3±2.7 vs 74.7±19.9 vs 99.3±0.7 %, p<0.001(vs NC/static), n=6]. Immunostaining for CD31 and SEM revealed sufficient coverage with endothelial cells in hL411/dynamic. Clot formation assay showed lower clot formation in hL411/dynamic group than in hL411/static (p=0.014) and mock without hL411 coating (p=0.006) (n=3).Conclusions:Surface modification of HHP-decellularized xenogeneic endothelial tissues combined with dynamic culture achieved sufficient ex vivoendothelialization accompanied with the prevention of clot formation which would promote the clinical use of endothelialized vascular grafts in the future.

Background Tricuspid valve regurgitation due to pacemaker leads is a well-known complication. Although some reports have suggested that pacemaker leads should be surgically explanted, strongly adhered leads cannot always be removed. The aim of this study was to describe our tricuspid valve repair techniques with pacemaker leads left in situ.Methods Our retrospective study investigated 6 consecutive patients who required tricuspid valve surgery for severe regurgitation induced by pacemaker leads.Results From the operative findings, we identified 3 patterns of tricuspid valve and pacemaker lead involvement. In 3 patients, the leads were caught in the chordae, in 2 patients, tricuspid regurgitation was caused by lead impingement on the septal leaflet, and in 3 patients, tricuspid valve leaflets had been perforated by the pacemaker leads. During surgery, all leads were left in situ after being separated from the leaflet or valvular apparatus. In addition, suture annuloplasty was performed for annular dilatation in all cases. In one patient, the lead was reaffixed to the annulus after the posterior leaflet was cut back towards the annulus, and the leaflet was then closed. There was one hospital death due to sepsis. The degree of tricuspid regurgitation was trivial in all surviving patients at discharge. During a mean follow-up of 21 months, one patient died from pneumonia 20 months after tricuspid valve repair.Conclusion In patients undergoing tricuspid valve surgery due to severe tricuspid regurgitation caused by pacemaker leads, the leads can be left in situ after proper repair with annuloplasty.

We present the case of a 19-day-old girl with incomplete atrioventricular septal defect, muscular ventricular septal defect, and severe left atrioventricular valve regurgitation. We attempted biventricular repair with left atrioventricular valve repair; however, we could not control the regurgitation. Moreover, the commercially available prosthetic valve was too large to implant. Thus we switched intraoperatively to a univentricular repair. We successfully performed patch closure of the left atrioventricular valve (Starnes procedure), Damus-Kaye-Stansel anastomosis, and a systemic-to-pulmonary artery shunt.

A 5-day-old girl who had pulmonary atresia with ventricular septal defect, patent ductus arteriosus, intralobar pulmonary sequestration of the right lower lobe, and retroesophageal innominate vein, was referred to our institution. We successfully performed a systemic-to-pulmonary shunt. During cardiopulmonary bypass, the arteries supplying the sequestrated lung were ligated above the diaphragm through the median sternotomy, without a thoracotomy. The postoperative course was uneventful, and we avoided the potential complications of lung infection and ventricular overload.

Although stem cell therapy is a promising strategy for cardiac restoration, the heterogeneity of transplanted cells has been hampering the precise understanding of the cellular and molecular mechanisms. Previously, we established a cardiovascular cell differentiation system from mouse pluripotent stem cells, in which cardiomyocytes (CMs), endothelial cells (ECs), and mural cells (MCs) can be systematically induced and purified. Combining this with cell sheet technology, we generated cardiac tissue sheets reassembled with defined cardiovascular populations. Here, we show the potentials and mechanisms of cardiac tissue sheet transplantation in cardiac function after myocardial infarction (MI). Transplantation of the cardiac tissue sheet to a rat MI model showed significant and sustained improvement of systolic function accompanied by neovascularization. Reduction of the infarct wall thinning and fibrotic length indicated the attenuation of left ventricular remodeling. Cell tracing with species‐specific fluorescent in situ hybridization after transplantation revealed a relatively early loss of transplanted cells and an increase in endogenous neovascularization in the proximity of the graft, suggesting an indirect angiogenic effect of cardiac tissue sheets rather than direct CM contributions. We prospectively dissected the functional mechanisms with cell type‐controlled sheet analyses. Sheet CMs were the main source of vascular endothelial growth factor. Transplantation of sheets lacking CMs resulted in the disappearance of neovascularization and subsequent functional improvement, indicating that the beneficial effects of the sheet were achieved by sheet CMs. ECs and MCs enhanced the sheet functions and structural integration. Supplying CMs to ischemic regions with cellular interaction could be a strategic key in future cardiac cell therapy. STEMCELLS2012;30:1196–1205

Prosthetic graft infection with methicillin-resistant Staphylococcus aureus (MRSA) is one of the most serious complications of cardiovascular surgery. Seeking to prevent graft infection, we evaluated the efficacy of a new biodegradable hydrogel glue (new-glue) composed of aldehyded dextran and epsilon-poly(L-lysine) which acts as a local sustained-release carrier of vancomycin. Rats (n=40) were implanted with 1-cm(2) Dacron grafts in the subcutaneous pockets. Groups (n=10 each) were as follows: no treatment (group A), topical vancomycin solution (group B), new-glue without vancomycin (group C) or new-glue containing 1 mg of vancomycin (group D). Twenty-four h after the implantation, 2.0x10(7) colony-forming units of MRSA was inoculated onto the graft surface. Seven days thereafter, the graft was sampled and cultured. The quantity of MRSA was significantly lower in group D than in the other groups (P<0.0001). About 95% of the total vancomycin was released from the new-glue over the 72 h experimental period, and the tissue concentration of vancomycin remained above the minimum inhibitory concentration for the MRSA strain throughout the experiment. This new vancomycin-containing glue effectively prevented prosthetic graft infection and thus may be a promising biodegradable drug vehicle.

Skeletal myoblast transplantation has been applied clinically for severe ischemic cardiomyopathy. Matrix metalloproteinase 1 (MMP-1) reduces fibrosis and prevents the progress of heart failure. We hypothesized that MMP-1 administration to the infarct area enhances the efficacy of skeletal myoblast transplantation. The controlled release of MMP-1 improved cardiac functions of rats with chronic myocardiac infarction with or without transplantation of skeletal myoblasts. Improvement in cardiac function and small fibrotic area inside the infarcted area were observed compared with those of myoblast transplantation. In conclusion, controlled release of MMP-1 was effective in cardioprotection in postmyocardial infarction although the combination with cell transplantation showed the similar effect.

We investigated the effects of partial left ventricular unloading on failing rat hearts by using heterotopic heart-lung transplantation model. Heart failure (HF) was induced in Lewis rats by ligating the left anterior descending artery. After four weeks, the infarcted hearts and lungs were harvested and transplanted into the recipient rats by anastomosing donor's ascending aorta to recipient's abdominal aorta. Therefore, coronary venous blood entered the left ventricle (LV) and LV was partially unloaded (HF-PU group). Normal and infarcted heart rats (HF group) without transplantation served as control animals. After two weeks' unloading, the infarcted LV in HF-PU group significantly decreased its weight and myocardial diameter compared with HF group and they were close to normal levels. Developed tension of posterior papillary muscle was significantly increased in HF-PU group compared with HF group. The mRNA expressions of brain natriuretic peptide (BNP), sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA2a), beta(1) and beta(2)-adrenergic receptors (beta(1) and beta(2)-AR) in LV tissue were almost normalized in HF-PU group. Partial left ventricular unloading regressed myocardial hypertrophy, reversed contractile dysfunction and normalized the mRNA (BNP, SERCA2a, beta(1) and beta(2)-AR) expressions of failing rat hearts.

When an arterial graft is used, reversible perfusion defects on single-photon emission tomography (SPET) perfusion images are occasionally observed early after coronary artery bypass graft surgery (CABG), owing to the restricted flow capacity. The purpose of this study was to determine whether the functional information obtained with electrocardiography (ECG)-gated perfusion SPET could be helpful in evaluating the effect of revascularization early after CABG. Twenty-three patients (18 men and 5 women, mean age 65±9 years) underwent stress/re-injection thallium-201 ECG-gated SPET before and 4 weeks after CABG (13 with exercise and 10 with dipyridamole). Patency of all grafts was confirmed by coronary angiography 1 month after CABG. Cardiac functional data including the left ventricular ejection fraction (LVEF) and the transient ischaemic dilatation (TID) ratio were analysed using a commercially available automated program. The conventional stress and re-injection tomograms were interpreted by means of a five-point scoring system in a nine-segment model. Stress-induced reversible 201Tl perfusion defects were present in 64% of the myocardial segments bypassed by patent arterial grafts, in contrast to 42% of the myocardial segments bypassed by patent venous grafts (?2=7.8,P=0.005). Of the 23 patients, 12 showed improvement in summed ischaemic scores (group 1), while 11 had no change or deterioration (group 2), although all grafts were patent on postoperative catheterization. The TID ratio improved in both group 1 and group 2 before and after CABG (1.14±0.13 vs 0.99±0.07,P=0.001 and 1.09±0.07 vs 0.94±0.05,P=0.002, respectively). However, LVEF did not significantly improve in group 1 or group 2 after CABG (42.5%±9.9% vs 47.5%±11.8%, and 52.1%±7.5% vs 53.1%±5.9%, respectively). Perfusion imaging or LVEF assessment is of limited value early after CABG. The TID ratio obtained with ECG-gated perfusion SPET may be a useful marker to evaluate the effect of revascularization early after surgery.

There is increasing evidence that catecholestrogens may contribute to the development of breast cancer. Specifically, inactivation of catecholestrogens may prevent the genesis and arrest the progression of the disease. Catechol-O-methyltransferase (COMT), Glutathione S-transferase (GST) M1 and GSTP1 are responsible for the detoxification of catecholestrogens, and are polymorphic in the human population. In this study, a PCR-based restriction fragment length polymorphism analysis was performed to determine genotypes of the COMT, GSTM1 and GSTP1 genes. We investigated the relationship between the germline polymorphism of these genes and clinico-pathological characteristics in 140 patients with breast cancer. Among 73 patients with the low activity COMT allele, 49 (67%) had regional lymph node metastasis. On the other hand, only 27 (40%) of 67 patients without the low activity allele had lymph node metastasis. The COMT genotype was significantly associated with clinical stage and the extent of regional lymph node metastasis of breast cancer (P<0.05). However, polymorphisms of the GSTM1 and GSTP1 gene were not associated with clinico-pathological factors. Our findings suggest that the allele encoding for low activity COMT may contribute to the progression of breast cancer.

Background:The optimum dose of granulocyte colony-stimulating factor (G-CSF) for peripheral blood stem cell (PBSC) mobilization after disease-oriented, conventional-dose chemotherapy remains unknown.Methods:A multicenter dose-finding study of glycosylated G-CSF (lenograstim) for the mobilization of PBSCs following adjuvant CAF chemotherapy (cyclophosphamide, doxorubicin and 5-fluorouracil) was performed in 38 patients with postoperative breast cancer. Each 10, ten and eight patients were sequentially allocated to one of the three dose groups (2,5 and 10 µg/kg, respectively) of lenograstim. Lenograstim was administered subcutaneously (s.c.) daily from day 8 to the day of the last apheresis and CD34+ cells and colony-forming units—granulocyte macrophage (CFU-GMs) in peripheral blood were measured serially. Additionally, 10 patients who received adjuvant CAF chemotherapy alone also participated in the study, as a control.Results:Lenograstim was well tolerated up to 10 µg/kg, except for one patient given 10 µg/kg who developed transient grade 3 hepatic enzyme elevation. The peak levels of CD34+ cells and CFU-GMs in peripheral blood showed dose—response relationships. The median peak CD34+ cells for the 0,2,5 and 10 Ég/kg dose groups were 5.4,34.3, 55.0 and 127.6 cells/Él, respectively, and those of CFU-GMs for the 0, 2, 5 and 10 Ég/kg dose groups were 0.01, 0.33,1.32 and 3.30 CFU-GMs/Él, respectively.Conclusions:Considering the previous reports suggesting that a pre-apheresis number of 40–50 CD34+ cells/µl in peripheral blood is highly predictive for achievement of more than 2.5 × 106CD34+ cells/kg in a standard apheresis procedure of 10 litres, the optimum dose of lenograstim for PBSC mobilization following CAF chemotherapy in patients with postoperative breast cancer is 5 µg/kg/day s.c.

Background:Liver metastasis from breast cancer has a poor prognosis. While there are some reports of good response rates of hepatic metastasis from breast cancer by hepatic intra-arterial infusion chemotherapy, no phase I study including pharmacokinetic analysis has been reported. We performed a phase I/II study of intra-arterial infusion chemotherapy using adriamycin and 5-fluorouracil to find the maximum tolerated dose and response rate in patients with advanced or recurrent breast cancer.Methods:A hepatic arterial catheter with an access port was inserted into the proper hepatic artery. Patients received 30 mg/m2adriamycin on days 1 and 8 and 100 mg/m25-fluorouracil at level 1, 200 mg/m2at level 2,300 mg/m2at level 3 and 400 mg/m2at level 4 continuously from day 1 through day 14 every 28 days. At least two cycles were required before evaluation. Twenty-eight patients were entered into this study and 26 patients were evaluable. Seventeen patients had hepatic metastasis only, although nine patients had additional metastasis to other sites.Results:Dose-limiting toxicity of thrombocytopenia and neurotoxicity occurred at level 4. Leukocytopenia (ECOG grade 3–4) was observed in five (19%), thrombocytopenia in three (12%) and anemia in two (8%) patients. There were 11 catheter-related complications which were not dose dependent. Seven out of 13 evaluable patients (54%) responded at level 3. The median duration of response was 5.8 months (range, 1–23+) and median survival was 25.3 months (range, 6.2–54.7+). Conclusion: Hepatic arterial infusion therapy appears to be safe and effective but catheterrelated complications must be overcome before starting a phase III trial.

Summary To study the effects of family history and reproductive, anthropometric, and dietary factors on the risk of breast cancer among low risk populations, we conducted a hospital-based case-control study involving 908 patients with breast cancer and their matched controls, in Japan. A positive family history of breast cancer significantly increased the risk of breast cancer (odds ratio = 1.52, 95% confidence interval: 1.14–2.03). The risk further increased with increasing number of family members affected. Obesity, single marital status, fewer births, a late childbirth, and less consumption of green-yellow vegetables and dairy products were also associated with an increased risk of breast cancer. These associations were independent in multivariate analyses. There was no increase in risk associated with consumption of high fat foods. When analyzed by menopausal status, the association with family history of breast cancer, especially in the first degree of relatives, was more evident for premenopausal breast cancer. The associations with obesity and lower consumption of dairy products were more pronounced for postmenopausal breast cancer, while those with lower parity and single marital status were stronger for premenopausal breast cancer.

To evaluate the effect of the oral fluoropyrimidines, tegafur and uracil (UFT) and 5-fluorouracil (5-FU), a pharmacodynamic analysis was conducted using a nude mouse system and patients. In the nude mouse system, UFT and 5-FU showed similar marginal effects against the human tumor xenograft Co-4, and the concentration of 5-FU in serum 1 h after the last administration being 0.04 µg/ml, which was assumed to be the minimum effective concentration. (MEC). Postoperative patients were subdivided into three groups, being: those who underwent subtotal gastrectomy and received UFT; those who underwent subtotal gastrectomy and received 5-FU; and those who underwent total gastrectomy and received UFT. In the UFT groups, the concentration of 5-FU in the portal and peripheral blood showed similar elimination in terms of the peak concentration (Cmax) and the area under the curve (AUC). In the 5-FU groups, the AUC and Cmax were significantly higher in portal blood than peripheral blood. The concentrations in the portal blood of the 5-FU group and in the portal and peripheral blood of the UFT group were significantly higher than the MEC (0.04 µg/ml). From these pharmacodymamic data, it was concluded that postoperative chemotherapy with oral fluoropyrimidines can achieve the MEC in portal and peripheral blood.

We have proposed the “vascular natriuretic peptide system”, in which C-type natriuretic peptide (CNP), the third member of the natriuretic peptide family, can control vascular tone and growth as an endothelium-derived relaxing peptide. We aimed at overexpression of CNP gene in vascular smooth muscle cells (SMCs) by adenovirus-mediated gene transfer to examine the growth characteristics of SMCs via the augmentation of cGMP production. Rat aortic SMCs infected with Ad.CNP, a replication-deficient adenovirus driving rat CNP cDNA, produced 162±55 fmol/mL of CNP, which was 4,000 times higher than that produced by endothelial cells. cGMP production was also augmented in Ad.CNP-infected SMCs (2200±270 fmol/104cells). Accordingly, significant growth inhibition was observed in SMCs infected with Ad.CNP. The flow cytometry analysis revealed that the population of the S and G2+M phases was reduced by 60% of the control in Ad.CNP-infected SMCs. The gene expression of ANP-B receptor, which is expressed abundantly in SMCs with the synthetic phenotype, was suppressed in Ad.CNP-infected SMCs, while the gene expression of ANP-A receptor, which is expressed predominantly in SMCs with the contractile phenotype, became detectable in Ad.CNP-infected SMCs. In addition, the gene expression of smooth muscle myosin heavy chain-2 (SM-2), which is the molecular marker of highly-differentiated SMCs, was also induced in Ad.CNP-treated SMCs. These results suggest that cGMP cascade activation induces re-differentiation of SMCs. The present study demonstrated that overexpression of CNP induced growth inhibition of SMCs at the G1 phase with possible alteration of the phenotype.

Thirty‐three out of46 strainsof Prevotella intermedia(Bacteroides intermedius) isolated from gingival deposits of patients with adult periodontitis possessed the ability to inhibit growth of P. intermediastrain ATCC 25611, and 6 of the 33 showed strong inhibitory activity. Thirteen isolates did not have the ability to inhibit the growth of P. intermediastrain ATCC 25611. An inhibition factor from one of the isolates, P. intermediaTH14 which had strong inhibition, was purified and characterized. Bacteriocin TH14 activity was effective against only P. intermediaATCC 25611 and Fusobacterium nucleatum, but not against other crevicular bacteria, including P. intermediaATCC 33563, Porphyromonas gingivalis(Bacteroides gingivalis), Capnocytophaga sputigenaand Actinobacillus actinomycetemcomitans. Streptococci, actinomyces, and veillonellas were not inhibited by bacteriocin TH14. Bacteriocin TH14 was heat sensitive, proteinaceous with molecular weight of 1,700 daltons and inhibited the growth of strain ATCC 25611 in 15 minutes. The growth inhibition of P. intermediaATCC 25611 by bacteriocin TH14 was found to be due to the bactericidal activity of the bacteriocin. J Periodontol 1991; 62:439–444.

Abstract:  Mammalian pancreatic ribonuclease (RNase) was conjugated chemically via a disulfide bond to human or murine epidermal growth factor (EGF). The conjugation between EGF and RNase was ascertained by SDS-PAGE using reduced and nonreduced conjugates. The EGF–RNase conjugate retained potent RNase activity and competed with 125I-EGF for binding to EGFR to the same extent as unconjugated EGF. Both the human and murine EGF–RNase conjugates showed dose-dependent cytotoxicity against EGFR- overexpressing A431 human squamous carcinoma cells with IC50 values of 3×10-7 M and 6×10-7 M, respectively, whereas free RNase had an IC50 of 10-4 M. Against the EGFR-deficient small-cell lung cancer cell line H69, the EGF–RNase conjugate had no cytotoxic effect. The Human EGF–RNase conjugate showed dose-dependent cytotoxicity against other squamous carcinoma cell lines (TE-5, TE-1) and breast cancer cell lines (BT-20, SK-BR-3, MCF-7) and the cytotoxicity of the conjugate correlated positively with the level of expression of EGFR by each cell line. An unconjugated mixture of EGF and RNase had no greater effect than RNase alone on any cell line. Excess free EGF blocked EGF–RNase conjugate cytotoxicity against A431 cells. These results suggest that the EGF–RNase conjugate may be a more effective anticancer agent with less immunogenicity than coventional chimeric toxins.

Abstract: The effect and mechanisms of 17β-estradiol (E2) on breast cancer cells were studiedin vivo andin vitro, using 5 human breast carcinomas serially transplanted into nude mice. These carcinoma strains consisted of 4 estrogen receptor (ER) positive tumors and 1 ER negative tumor. Mice bearing these tumors were treated with an intramuscular injection of E2 at a dosage of 50 mg/kg and the tumor doubling time (Td) was calculated in days. The tumor growth was significantly stimulated by E2 in 3 out of the 4 ER positive tumors, the Td of the E2 treated groups being 17.6 days for MCF-7 (control: −17.8 days), 12.8 days for R-27 (control: −12.5 days∼14.5 days) and 10.4 days for Br-10 (control: 14.5 days), however, in the T-61 tumor, the growth was inhibited by E2 in a dose dependent manner. In the case of the ER-negative MX-1 tumor, the tumor cell growth was not affected by E2. Discrepancies between the effects of E2 on ER-positive tumors were further analyzed by examining the steroid hormone receptor status and conductingin vitro growth studies.In vitro clonogenic cell assay reproduced the antitumor activity of E2, indicating that E2 directly inhibits part of the cell growth of T-61 tumors. The above results suggest that this experimental system provides a useful tool for analyzing the mechanism of estrogen in breast cancer and that the clonogenic assay using ER positive specimens can help to identify breast cancers sensitive to estrogen therapy.

In order to investigate the influence of estrogenmetabolism on human breast cancer, estradiol 2- and16α-hydroxylase (2- and 16α-OHase) activities were determined inthe microsomal fractions of cancer tissues by usingreverse phase HPLC. 2-OHase activity was detected inmost cancer tissues and noncancerous tissues, but theactivity was significantly lower in cancer tissues thanin the paired noncancerous tissues (0.01 < p< 0.02). Interestingly the patients without lymph nodemetastasis had significantly higher 2-OHase activity in cancertissues than those with lymph node metastasis (0.02< p < 0.05). No correlation was observedbetween ER status and 2-OHase activity in cancertissues. On the other hand, 16α-OHase activity wasdetected only in one third of the breastcancer tissues examined. The activity was not significantlydifferent from that in noncancerous tissues, although itwas relatively higher in ER-positive cancer tissues whencompared with that in ER-negative ones (0.05

Adriamycin (ADM) was chemically conjugated to a murine monoclonal antibody, A0011, which recognizes the c-erbB-2 product, via a disulfide bond using N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP) and 2-iminothiolane (2-IT). The molar ratio of ADM to the monoclonal antibody ranged from 15:1 to 25:1 and enzyme-linked immunosorbent assay (ELISA) showed that the binding activity of the conjugate was almost retained. We compared the efficacy of A0011 alone, ADM alone, the A0011-ADM conjugate, and a nonspecific murine IgM-ADM conjugate, against the human breast cancer cell lines SK-BR-3, MDA-MB-361, MCF-7, and BT-20. The A0011-ADM conjugate was observed to be ten times more cytotoxic to the cell lines overexpressing the c-erbB-2 product, namely, SK-BR-3 and MDA-MB-361, than free ADM, but it showed weak cytotoxicity against the cell lines with a low level of c-erbB-2 product expression, namely, MCF-7 and BT-20. However, free A0011 and nonspecific murine IgM-ADM conjugate showed no cytotoxicity toward any of the four cell lines, while the addition of a tenfold molar excess of A0011 inhibited conjugate cytotoxicity. These data suggest that conjugate cytotoxicity is antibody-mediated. Moreover, conjugate cytotoxicity at 10-6M was correlated with antigen volume, and the data were fitted to the regression equation y=-11.63logX+116.38 where the correlation coefficient =0.950. Our results indicate that targeting therapy aiming at the c-erbB-2 product may be useful in the treatment of breast cancers overexpressing the c-erbB-2 product.

Introduction:We have been investigating cardiac regenerative medicine using human induced pluripotent stem cells (hiPSCs) with our robust differentiation system toward cardiovascular populations (cardiomyocytes [CMs]/ endothelial cells [ECs]/ vascular mural cells [MCs]), and a formulation of 3-dimensional structure (Human iPSC-derived Cardiac Tissue; HiCT) from the cardiovascular cells.Hypothesis:The HiCTs hold therapeutic potential for animal heart disease models.Methods:We conducted preclinical efficacy studies to test therapeutic effects of HiCTs. We simultaneously induced cardiovascular cells from hiPSCs with our monolayer culture method, then generated hiPSC-derived cardiac tissue sheets (CTSs) including CMs, ECs and MCs with temperature-responsive culture dishes. We generated 5-layered HiCTs with high viability (live cells: 99.4?0.1 %) by stacking 5 CTSs with insertion of gelatin hydrogel microspheres to overcome the stacking limitation of hypoxia avoiding central necrosis. We transplanted the HiCTs onto a ?-sarcoglycan-deficient dilated cardiomyopathy (DCM) hamster model and a porcine myocardial infarction (MI) model.Results:Transplantation (Tx) of HiCTs significantly alleviated left ventricular dilation (Sham vs Tx, end-diastolic diameter: 0.75?0.02 vs 0.63?0.03 cm, p<0.001) and functional deterioration [ejection fraction (EF): 40.9?1.6 vs 58.2?1.9 %, p<0.001] of the DCM hamsters at 3 months after transplantation. Sirius-red staining showed lower fibrotic area (17.5?1.1 vs 10.4?0.8 %, p<0.001) which was also confirmed by synchrotron radiation-based imaging. TUNEL staining indicated significantly fewer apoptotic cells compared to those in sham controls (p<0.01). The transplantation of clinical-scale HiCTs onto a porcine MI model resulted in significantly higher EF compared to those in sham controls (55.7 ? 24.2 vs 79.4 ? 6.8 %, p<0.001), reaching to comparable level as those before MI induction. Vascular networks among engrafted HiCTs from genetically labelled host pig hearts were confirmed by light-sheet microscopy for samples after tissue clearing.Conclusions:Here we validated the functional benefits of HiCT transplantation therapy onto diseased animal hearts.

Introduction:Acute lung injury associated with systemic inflammatory response syndrome leads to respiratory failure, a crucial complication after cardiac surgery using cardiopulmonary bypass (CPB). Recombinant human soluble thrombomodulin (rTM) consists of three domains (D123) and is reported to possess anti-inflammatory effects through binding to high mobility group box 1 (HMGB1) as a cytokine mediator of inflammation. The binding site of HMGB1 is located at the N-terminal lectin-like domain (D1). Simultaneously, rTM has anticoagulant activities through binding to thrombin at epidermal growth factor-like domain (D2) which may exacerbate coagulopathy by CPB.Hypothesis:D1 domain of rTM has the potential against CPB-mediated acute lung injury free from anticoagulant effects in a rat CPB model.Methods:Fifteen male Sprague-Dawley rats were divided into 3 groups: control (CPB alone, n=5), D1 (CPB + D1, n=4), and D123 (CPB+D123, n=6). A bolus of D123 (3 mg/kg) and D1 (equivalent to 3 mg/kg of D123) was administered to each group of rats 30 minutes before CPB establishment. CPB was conducted for 90 minutes. Blood samples are collected at just before administering reagent, during and after CPB, respectively. Blood coagulability was assessed by an coagulation function analyzer. Lung samples are collected at 1 hour after the end of CPB for histological analyses.Results:Only D123 group exhibited significantly prolonged glass beads activated clotting time with heparinase (H-gbACT) after CPB compared to those before CPB (83.5 ? 1.9 vs. 69.0 ? 2.6 sec, p = 0.026 [after CPB vs. before CPB]), while D1 group did not show prolonged H-gbACT (65.3 ? 5.5 vs. 71.3 ? 3.6 sec [after CPB vs. before CPB]) indicating the absence of anticoagulant activities of D1. Blood HMGB1 level tended to be lower in both D1 and D123 groups (14.1 ? 0.9 vs. 10.6 ? 1.7 vs. 10.3 ? 1.4 ng/ml [Control vs. D1 vs. D123]). Histological studies revealed less congestion, edema, inflammation, and hemorrhage in both D1 and D123 groups compared to those in control group indicating protective effects of both D1 and D123 against acute lung injury mediated by CPB.Conclusions:These results indicated that the N-terminal lectin-like domain of rTM reduces acute lung injury without anticoagulant effects in a rat CPB model.

Introduction:Autologous vascular grafts are widely used in revascularization surgeries for small caliber targets. However, the availability of autologous conduits may be limited due to prior surgeries and the quality of vessels. Decellularized vascular grafts from animals potentially substitute for autologous vascular grafts. Decellularization with high hydrostatic pressure (HHP) has been reported to preserve the structure and composition of extracellular matrix (ECM) highly, which is advantageous for recellularization and vascular remodeling after implantation.Hypothesis:HHP decellularized vascular grafts retain capacity for recellularization with fair patency in a xeno-implantation animal model.Methods:Bovine dorsalis pedis arteries were decellularized by HHP method. We interposed porcine right carotid arteries with the bovine decellularized grafts (N=5). No anticoagulants or anti-platelet agents are administrated during the observation period. Ultrasonography, angiography and intravascular ultrasound were used to evaluate the patency and morphology of the grafts at 2 and 4 weeks after implantation, respectively. Grafts were explanted at 4weeks after implantation and subjected to histological analyses.Results:All 5 grafts were patent at 4 weeks after implantation with mild stenosis close to the anastomosis site. Neither aneurysmal formation nor massive thrombosis was observed. Elastica van Gieson and Sirius-red stains revealed fair preservation of ECM proteins including elastin and collagen after implantation. Inner lumen of grafts was thoroughly covered with von Willebrand factor-positive endothelial cell layer. Migration of alpha-smooth muscle actin-positive smooth muscle cells into tunica media was observed.Conclusions:Xenogeneic HHP decellularized graft provided feasible capacity for recellularization and vascular remodeling without thrombogenicity. HHP decellularized vascular grafts may be utilized as grafts for revascularization surgeries for small caliber arteries.

Two monoclonal antibodies, 17(3)9 and 36(79)4, were prepared against nitrate reductase from Spinacia oleracea L. leaves. An enzyme-linked immunosorbent assay showed that 17(3)9, but not 36(79)4, reacted more strongly to heat-denatured than native antigen. These antibodies inhibited NADH-nitrate reductase as well as its various partial activities including reduced methyl vilogen-nitrate reductase, reduced flavin mononucleotide-nitrate reductase and NADH-cytochrome c reductase activities, but not NADH-ferricyanide reductase activity. Immunoblotting after electrophoretic separation of nitrate reductase fragments obtained by Staphyrococcus aureus V8 protease digestion of native enzyme revealed that the two monoclonal antibodies bind to different epitopes located on the 28 kDa of the NADH-ferricyanide reductase domain.

Adherence of 3H-labelled cells of Streptococcus gordonii and Streptococcus milleri to artificial pellicles prepared from saliva supplemented with glucosyltransferases from mutans streptococci was examined using a new assay for sucrose-dependent cell-to-pellicle attachment. Results indicate that S. gordonii, but not S. milleri, could attach tightly to hydroxylapatite surfaces through de novo glucan synthesis by mutans streptococcal glucosyltransferases present in the experimental salivary pellicles.

Anomalous increase of the effective anisotropy field change ?(HK-4?MS) has been found when magnetic bubble garnet films were heavily implanted with Ne+/He+and subsequently annealed at high temperatures (350°C). With annealing, ?(HK-4?MS) increases with annealing time, reaches a maximum value and then decreases. ?(HK-4?MS) vs. strain (?a/a) does not follow the curve for as-implanted films. ?(HK-4?MS) has reached up to 3700 Oe which is 1200 Oe larger than that expected for as-implanted films (2500 Oe). This anomalous increase has been observed only for films whose initial strain is more than about 1.5%; the effect has not been observed in heavily H+2implanted films.