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A multicenter, randomized, prospective trial was conducted to determine the clinical efficacy of ticlopidine, an antiplatelet agent, in the prevention of graft occlusion following vascular reconstructive surgery of the lower extremity for arteriosclerosis obliterans. Ticlopidine was administered for two years to one group of patients (112 reconstructed arterial segments) while a control group received no antithrombotic therapy (108 segments). There was no significant difference between the treated and control groups in the overall cumulative patency rates for both suprainguinal and infrainguinal reconstructions. In the treated group, however, there was a significant reduction of occlusions in pa tients with rest pain or ischemic gangrene (p < 0.1) and in those with hyperlipi demia (p < 0.05). Ticlopidine-treated patients with infrainguinal arterial reconstructions developed more occlusions than controls between three and twelve months after surgery, but the controls had more occlusions both immedi ately after surgery and in the second postoperative year. It appeared, therefore, that ticlopidine reduced graft failure due to neointimal fibrous hyperplasia or progression of the underlying condition but caused an adverse effect by delaying graft healing. The clinical usefulness of antiplatelet therapy is limited, but it may be effective in the prevention of graft occlusion when administered to the selected patient groups above mentioned. Controversy about antiplatelet ther apy and future research on adjuvant drug therapy after arterial reconstruction are discussed. [ABSTRACT FROM PUBLISHER]

Escherichia coli is an important fecal indicator bacterium that is used to evaluate the microbiological quality of water. The Colilert-18 (Quanti-Tray/2000) is a widely used, rapid, and simple quantitative method for detecting E. coli in drinking water, bathing water, and wastewater. However, Colilert-18 method is less reliable for seawater; false positives are often caused by halophilic bacteria such as Vibrio. While false positives can be avoided by diluting the sample by 10 times or more, the resulting decrease in detection limit makes it difficult to quantify E. coli in seawater. In this study, we combined cake filtration, using hydroxyapatite powder, with the Colilert-18 method to remove salinity without diluting the water sample. When quantifying E. coli in river water, the E. coli concentration obtained from the cake filtration/Colilert-18 method showed a high quantitative value of 90% or more, on average, compared to the concentration obtained with the original Colilert-18 method. The E. coli concentrations in seawater determined using the developed method were similar to those determined using the modified m-TEC method, with no false positives. Highly reliable quantitative values can be obtained using the proposed method because it is possible to measure 100 times as much sample compared to the dilution method. Thus, the developed method is expected to be a powerful tool that can eliminate the problem of false positives., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Kudoa septempunctata is the causative agent of a food-borne disease associated with the ingestion of raw olive flounder. As the current qRT-PCR method for its detection is time-consuming, a rapid and simple method is required. Recently, a new real-time loop-mediated isothermal amplification (LAMP) method and an immunochromatography method, whose sensitivities are intended to be compatible with that designated in the official analytical method (10(5) spores/g olive flounder), have been developed. To validate these new methods, we performed an inter-laboratory study across seven laboratories. Both methods could not detect less than 10(4) spores/g; however, these methods were able to detect more than 10(5) spores/g in olive flounder samples. These results demonstrated that the sensitivities of these methods were compatible with the designated level in the official analytical method. We concluded that these new methods were acceptable as the screening methods for K. septempunctata.

The heterologous expression of the laccase gene from Trametes versicolor and Gaeumannomyces graminis was evaluated in the yeasts Pichia pastoris and Cryptococcus sp. S-2. The expression levels of both laccase genes in Cryptococcus sp. S-2 were considerably higher than those in P. pastoris. The codon usage of Cryptococcus sp. S-2 as well as the GC content were similar to those of T. versicolor and G. graminis. These results suggest that using a host with a similar codon usage for the expressed gene may improve protein expression. The use of Cryptococcus sp. S-2 as a host may be advantageous for the heterologous expression of genes with high GC content. Moreover, this yeast provides the same advantages as P. pastoris for the production of recombinant proteins, such as growth on minimal medium, capacity for high-density growth during fermentation, and capability for post-translational modifications. Therefore, we propose that Cryptococcus sp. S-2 be used as an expression host to improve enzyme production levels when other hosts have not yielded good results., (Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Background and Objective: To quantitatively estimate tongue function, we developed a handy device for intraoral pressure measurement. The objective of this study was to assess maximum voluntary tongue pressure (MVTP) in Japanese frail elderly persons receiving nursing care services., Materials and Methods: The study included 42 men and 87 women, aged 58-100 years. To record MVTP, the participants were asked to compress the balloon (diameter: 18 mm) of the disposable intraoral pressure probe onto their palates for 7 s using the maximum voluntary effort of the tongue. Pressures were recorded three times at 1 min intervals., Results: Maximum voluntary tongue pressure was successfully measured in 111 persons. Mean (standard deviation) MVTP was 18 (12) kPa, with a range of 0-63 kPa. The remaining 18 persons could not accurately follow our instructions and MVTP could not be measured., Conclusion: In comparison with the reported standard value using the same device, the frail elderly persons included in our study were found to exert less MVTP than healthy dentate individuals. These results suggest the need for proper quantitative evaluation of oral function, including tongue capacity, at nursing care facilities., (© 2012 The Gerodontology Society and John Wiley & Sons A/S.)

Yeast host-vector systems have been very successful in expressing recombinant proteins. However, because there are some proteins that cannot be expressed with existing systems, there is a need for new yeast expression systems. Here we describe a new host-vector system based on the basidiomycetous yeast Cryptococcus sp. strain S-2 (S-2). Two advantages of S-2 are that it naturally produces some very useful enzymes, so it would be a good system for expressing multiple copies of some of its genes, and that, it is a nonhazardous species. The orotate phosphoribosyltransferase (OPRTase, EC 2.4.2.10) gene (URA5) was selected as a selectable marker for transformation in the new host-vector system. URA5 was isolated and introduced into a uracil auxotroph of S-2 by electroporation. To demonstrate the S-2 system, we selected one of its unique enzymes, a plastic-degrading cutinase-like enzyme (CLE). We were able to insert multiple copies of the CLE gene (CLE1) into the chromosomes in a high fraction of the targeted cells. Under optimal conditions, one transformant exhibited 3.5 times higher CLE activity than the wild type. Expression vectors, including an inducible promoter (the promoter for the xylanase or α-amylase gene), were constructed for recombinant protein production, and green fluorescent protein was expressed under the control of these promoters. The xylanase promoter was more tightly controlled. Furthermore, putting CLE1 under the control of the xylanase promoter, which is induced by xylose, increased CLE activity of the culture medium to approximately 15 times greater than that of the wild type.

Periodontal regeneration has been challenged with chemical reagents and/or biological approaches, however, there is still no sufficient technique that can regenerate complete periodontium, including alveolar bone, cementum, and well-oriented collagen fibers. The purpose of this study was to examine multi-layered sheets of periodontal ligament (PDL)-derived cells for periodontal regeneration. Canine PDL cells were isolated enzymatically and expanded in vitro. The cell population contained cells capable of making single cell-derived colonies at an approximately 20% frequency. Expression of mRNA of periodontal marker genes, S100 calcium binding protein A4 and periostin, was observed. Alkaline phosphatase activity and gene expression of both osteoblastic/cementoblastic and periodontal markers were upregulated by osteoinductive medium. Then, three-layered PDL cell sheets supported with woven polyglycolic acid were transplanted to dental root surfaces having three-wall periodontal defects in an autologous manner, and bone defects were filled with porous beta-tricalcium phosphate. Cell sheet transplantation regenerated both new bone and cementum connecting with well-oriented collagen fibers, while only limited bone regeneration was observed in control group where cell sheet transplantation was eliminated. These results suggest that PDL cells have multiple differentiation properties to regenerate periodontal tissues comprising hard and soft tissues. PDL cell sheet transplantation should prove useful for periodontal regeneration in clinical settings.

Objective: The ultimate goal of periodontal treatment is to regenerate the damaged periodontal support. Although periodontal ligament (PDL) cells are essential for periodontal regeneration, few studies have reported the transplantation of periodontal ligament cells to periodontal defects. We developed a new method to apply periodontal ligament cells as a sheet to the defect. The aim of this study was to investigate the periodontal healing after application of the periodontal ligament cell sheet in beagle dogs., Methods: Autologous periodontal ligament cells were obtained from extracted premolars of each beagle dog. Periodontal ligament cell sheets were fabricated using a temperature-responsive cell culture dish. Dehiscence defects were surgically created on the buccal surface of the mesial roots of bilateral mandibular first molars of each dog. In the experimental group (five defects), periodontal ligament cell sheet with reinforced hyaluronic acid carrier was applied to the defect. Only the hyaluronic acid carrier was applied to the contralateral side as a control (five defects). Eight weeks after surgery, the animals were sacrificed and decalcified specimens were prepared. Healing of the periodontal defects was evaluated histologically and histometrically., Results: No clinical signs of inflammation or recession of gingiva were observed in both experimental and control groups. In the experimental group, periodontal tissue healing with bone, periodontal ligament and cementum formation was observed in three out of five defects. In the control group, such periodontal tissue formation was not observed except in one defect. Histometric analysis revealed that the formation of new cementum in the experimental group was significantly higher than that in the control group., Conclusion: The periodontal ligament cell sheet has a potential to regenerate periodontal tissue and may become a novel regenerative therapy.

Background: A novel injectable, fast setting calcium phosphate cement (CPC) is currently used in orthopedic therapy for bone fractures. This study evaluated the possibility of applying this cement to healing periodontal defects., Methods: Fenestrations and 3-walled periodontal defects were surgically created on bilateral first molars and canines in 5 beagle dogs. CPC was applied to the defects on one side of the mandible. Untreated defects on the contralateral side served as controls. CPC was applied to all defects in the maxilla. Twelve weeks after surgery, the animals were sacrificed and decalcified and undecalcified specimens were prepared. Periodontal tissue healing was evaluated histologically and histometrically under a light microscope., Results: Healing of periodontal tissues in terms of bone and cementum formation was consistently observed in the CPC-applied sites. CPC was partly replaced by new bone. The residual CPC appeared detached from the denuded root surface. New cementum and periodontal ligament-like tissue were observed between the detached CPC and root surface. No unfavorable reaction was noted in the CPC-applied sites. No statistically significant difference was noted in the experimental or control sites under histometric analysis., Conclusions: Although there were no statistically significant differences between the 2 treatment groups, histological observation indicated that CPC seemed to act as a scaffold for bone formation and provided histocompatible healing of periodontal tissues in this study. This cement might be applicable to periodontal therapy; however, further investigations are required.

A 65-year-old male was admitted to our hospital because of bilateral giant bullae and right side pneumothorax. He had had a myocardial infarction and had been receiving medications. Considering his age, ischemic heart disease and hypertension, we excised the bullae in two operations. The first operation was open chest bullectomy of a giant bulla and small bulla on the right side. After this procedure, his clinical course was uneventful. In the second operation, thoracoscopic excision of the left side giant bulla was performed using only one endoscopic stapler by rotating each side of the bulla and making a clear excision line between the bulla and normal lung. His post operative course was excellent and he was discharged from our hospital on the 15th day after the second operation.

Nonpenetrating blunt trauma causing pulmonary pseudocyst is very rare. A 25-year-old young male suffered from motor vehicle accident. He was transferred to our hospital with severe hemoptysis. Immediately he was intubated and kept under mechanical ventilation. A chest X-ray film and CT-scan revealed a marked cavity lesion surrounded by an extensive infiltrate and alveolar air consolidation. Ordinary, it has benign clinical courses by conservative management. However, he was suffered massive and continuous bleeding from endobronchial tube. Therefore, one lung ventilation using a uni-vent tube blocker was performed. After twelve hours blockade, the cavity lesion was enlarged and trans-bronchial bleeding was continued severely. Emergency right lower lobectomy was carried out successfully.

A case of cystic adventitial disease (CAD) of the popliteal artery with intermittent claudication in the left calf is reported. This patient was first treated by total excision of the cyst and the involved artery followed by graft interposition with an autogenous saphenous vein. Recurrence of CAD in the interposed vein graft was noted after 6 months, and excision of the cyst with the involved graft and graft interposition with an autogenous saphenous vein was again required. We consider that the definitive evidence from early recurrence in the interposed vein graft shown in this case will close the discussion of the cause of CAD. The cystic lesion is thought by some to originate from the adventitia; traumatic, embryologic, or systemic abnormality theories were proposed. We assume that the mucin-secreting synovial cells originating from the neighboring joint capsule, tendon sheath, or, in some cases, from the ganglion itself, directly invade the adventitia through any rough or injured crack caused by trauma to the adventitia or simply attach to and encircle the adventitia. Total resection of the lesion with grafting is the recommended treatment for CAD because of the excellent results. However, as shown in our case, complete removal of the synovial cells even by resection technique may be difficult; therefore intensive follow-up is fundamentally necessary in this disease.

A case of a 51-year-old male with atresia of the right coronary artery and annulo-aortic ectasia is described. He presented with heart failure and underwent open-heart surgery. Preoperative aortography did not show the ostium of the right coronary artery. At surgery the right coronary ostium was not found on the intimal surface of the aneurismal aortic wall. A fine cord was attached to the adventitial surface of the anterior aortic root. A modified Bentall's procedure was performed to reconstruct the left coronary artery alone. The post-operative course was uneventful. It is speculated that the hypoplastic ostium of the right coronary artery was pulled up and formed a flap-like closure with increased expansion of the aorta.

Although the measurement of serum haptoglobin (S-Hp) is of great use for evaluation of intravascular hemolysis, it is not applicable in patients with mechanical prosthetic valves because S-Hp is virtually absent. We administered haptoglobin preparation to 10 patients with Björk-Shiley mitral prosthetic valves and 10 patients with the same aortic prosthetic valves. Serum haptoglobin levels were measured periodically afterwards. The maximum haptoglobin levels (Hp (max)), serum, haptoglobin reducing rate ((Hp-delta Hp)/delta t) and expected haptoglobin disappearing time (hours) were obtained from the subsequent samples. The screening studies which were performed at the same time were not predictors of difference in the 2 groups. On the other hand, serum haptoglobin reducing rate and expected haptoglobin disappearing time indicated that hemolysis is higher in patients with an aortic prosthetic valve than with a mitral prosthetic valve. This haptoglobin administration test seems to be useful for the comparative examination of the intravascular hemolysis caused by the difference in the position of the prosthetic valve.

A case of acute aortic dissection Stanford Type A occurring in a patient with Marfan's syndrome, complicated with aortic regurgitation and severe pectus excavatum, is reported with successful surgical correction. The patient was a 33-year-old woman, who suffered from severe back pain riding to work. An emergency operation consisting of Cabrol's procedure for aortic dissection Stanford Type A and sternal turnover for pectus excavatum was performed simultaneously. Because of the postoperative mediastinal bleeding the implanted sterno-costal complex was removed on the second POD and the prosthetic sternum was reimplanted on the tenth POD. The two-stage operation seems preferable in such an emergency case to avoid postoperative bleeding and infection.

A case of quadricuspid aortic valve is reported in a patient with coronary artery disease and abdominal aortic aneurysms. A 54-year-old male who had undergone aortic replacement because of abdominal aortic aneurysms three years before presentation was readmitted due to complaints of angina pectoris and palpitations. Aortography and coronary arteriography revealed severe aortic regurgitation and proximal occlusion of LAD and RCA. Surgical correction consisted of aortic valve replacement with a Björk-Shilely valve and coronary revascularization of LAD. During the operation, a quadricuspid aortic valve with one smaller and three larger cusps that showed mild myxomatous degeneration without dystrophic calcification and normal coronary arterial orifices were noted. Accordingly, severe aortic regurgitation may have resulted from the dysfunction of congenital malformed cusps and acquired sclerotic coronary disease was the main cause of the chest pain.

We report a successful surgical treatment of acute dissecting aneurysm of the ascending aorta associated with annuloaortic ectasia. The patient was a 29-year-old male, who was admitted to the hospital 5 hours after the onset of anterior chest pain. Aortography revealed acute dissection of the ascending aorta (DeBakery type I) with AAE. The intimal tear surrounding the right coronary ostium was found. The procedure between the right coronary artery and aorta (replaced artificial graft) using saphenous vein graft was also required in addition to Bentall's operation. Postoperative studies revealed disappearance of the dissection, patency of the bypass graft. The postoperative course was uneventful and the patient is in active life two years after operation.

We have recently performed a successful repair of a total right anomalous pulmonary venous connection to the coronary sinus associated with mitral and tricuspid regurgitation. A 54-year-old female was inaccurately diagnosed as atrial septal defect associated with mitral and tricuspid regurgitation preoperatively. Operation revealed that all the right pulmonary veins connected to the coronary sinus. Mitral regurgitation which might be caused by distortion of the anterior leaflet was repaired by McGoon's technique. A Carpentire-Edwards ring was employed for the tricuspid annuloplasty. Repair of the atrial septal defect was undertaken with a pericardial septation patch which was sutured in place. The roof of the coronary sinus was widened by cutback, so that blood from the pulmonary veins could drain into the left atrium. The postoperative course was uneventful, except for a supraventricular arrhythmia.

A 61-year-old man with Campylobacter fetus infection of an abdominal aortic aneurysm treated surgically is presented herein, the fifth survival case reported in the literature. Fever and back pain preceded the enlargement of atherosclerotic abdominal aortic aneurysm. The patient tolerated satisfactorily total excision of the aneurysm followed by axillo-femoral prosthetic bypass. Antibiotic therapy consisted of intravenous infusion of fosfomycin and gentamicin and oral administration of minocycline. The organism cultured from the aneurysmal wall and intraluminal thrombi was identified as Campylobacter fetus from its typical characteristics. It is concluded that this organism should be considered in all cases of infected aneurysm in elderly or debilitated patients.

A knitted Dacron velour externally supported prosthesis was broken in two pieces at the supported portion 3 years after femoroposterior tibial bypass grafting. This type of graft problem has never been reported. Details of the clinical course, angiogram, CT scan, and scanning electron micrograph of a portion of the graft are shown. The externally supported Dacron prosthesis broke into two pieces along the broken coil of the middle portion. A possible cause of this failure may have been due to damage of the Dacron fibers in the process of heat fusing the coil. This presentation emphasizes the possibility of this rare externally supported graft complication, and the necessity for careful follow-up not only of the graft patency but also for the mechanical defects of the implanted externally supported Dacron graft itself.

One stage operation was successfully performed on a 32-year-old man with annulo-aortic ectasia (AAE) and pectus excavatum caused by Marfan's syndrome. In the Bentall's operation for AAE, median sternotomy is usually carried out. In this case it was difficult to obtain a favorable viewing field due to an extremely deformed sternum and significant displacement of the heart into the left thoracic cavity. By removing the sternum en bloc, as a cost-sterno complex, the surgery was performed safely with an excellent operative field. In this surgery combined with the sternal turnover, the amount of intra and post operative bleeding was the same as occurs in other open heart surgeries.

A 31-year-old male was diagnosed to have malignant thymoma with superior vena cava occlusion. At operation, we performed extended resection of tumor and superior vena cava, and replacement with ringed PTFE graft. Histological diagnosis was nodular sclerotic type of Hodgkin's disease of thymus. He received radiation with Liniac and chemotherapy post operatively, and post operative venography revealed graft patent. He is doing well 3 years after surgery.

A 11-year-old boy with supravalvular aortic stenosis was treated by extended aortoplasty (Doty's operation) with excellent results. The extended aortoplasty provided satisfactory relief of aortic stenosis and improved function of the aortic valve.

A thirteen-year-old asymptomatic boy was referred to our hospital because of a cardiac murmur detected on a routine physical examination at school. A plane chest radiography showed increased vascular markings in the left lower field. An Aortogram revealed a large artery arising from the descending aorta and suppling the left posterior basal segment which had no pulmonary arteries. A bronchogram showed no abnormal findings in the bronchial tree. A clinical diagnosis of systemic origin of an aberrant artery to the basal segments of the left lung was made, and left lower lobectomy was performed successfully. We discussed the difference of this anomaly from pulmonary sequestration as well as the surgical procedure.