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We report a rare case of retrograde Stanford type A aortic dissection after endovascular repair for complicated Stanford type B aortic dissection. A 45-year-old man presented with a sudden onset of back pain and was transferred to our hospital. Computed tomography demonstrated acute Stanford type B aortic dissection with lower limb ischemia. Emergency endovascular surgery was planned for repair of the Stanford type B aortic dissection. The patient suddenly developed recurrent chest pain 10 days after the initial procedure. Computed tomography revealed retrograde Stanford type A aortic dissection involving the ascending aorta and aortic arch. The patient underwent a successful emergency total aortic arch replacement.

The Lachman test is considered to be a reliable physical examination for anterior cruciate ligament (ACL) injury. Patients with a damaged ACL demonstrate a soft endpoint feeling. However, examiners judge the soft and hard endpoints subjectively. The purpose of our study was to confirm objective performance of the Lachman test using joint auscultation. Human and porcine knee joints were examined. Knee joint sound during the Lachman test (Lachman sound) was analyzed by fast Fourier transformation. As quantitative indices of Lachman sound, the peak sound as the maximum relative amplitude (acoustic pressure) and its frequency were used. The mean Lachman peak sound for healthy volunteer knees was 86.9 ± 12.9 Hz in frequency and -40 ± 2.5 dB in acoustic pressure. The mean Lachman peak sound for intact porcine knees was 84.1 ± 9.4 Hz and -40.5 ± 1.7 dB. Porcine knees with ACL deficiency had a soft endpoint feeling during the Lachman test. The Lachman peak sounds of porcine knees with ACL deficiency were dispersed into four distinct groups, with center frequencies of around 40, 160, 450, and 1600. The Lachman peak sound was capable of assessing soft and hard endpoints of the Lachman test objectively.

Background Because increased age is a strong independent predictor of mortality and morbidity, surgery for octogenarians with Stanford type A aortic dissection (AAD) may be avoided.Methods and Results:From 2005 to 2015, 158 patients underwent surgical repair for AAD via a median sternotomy. We compared 24 (15.2%) octogenarians (83±3 years) with 134 (84.8%) patients aged ≤79 years (62±13 years), based on retrospectively collected clinical data. Octogenarians were predominantly female (79.2% vs. 44.8%, P=0.0033). Ascending aortic replacement was more frequently performed in the octogenarians (95.8% vs. 65.7%, P=0.0015) and total arch replacement in the younger patients (4.2% vs. 26.9%, P=0.0165). There were 14 hospital deaths among the younger patients, none among the octogenarians (0% vs. 10.4%, P=0.1303), and major morbidity rates were comparable. There were 3 late deaths among the octogenarians and 9 deaths among the younger patients. The respective 1-, 3-, and 5-year survival rates were 94.4%, 81.5%, and 81.5% in the octogenarians and 86.9%, 85.6%, and 83.9% in the younger patients, with no significant differences. Conclusions Surgical repair for AAD in octogenarians showed favorable results when compared with a younger patient cohort, with low hospital mortality rate and excellent late outcomes. Therefore, this technique should not be disregarded just because the patient is an octogenarian. (Circ J 2016; 80: 2468-2472).

Highlights • The following two points are the subject of this case report. First, autologous blood collection can be a risk factor of nonocclusive mesenteric ischemia (NOMI). Second, delayed intestinal stenosis after the recovery from NOMI may require surgical intervention in some cases. As far as the authors know, there is no report about NOMI associated with autologous blood collection. In addition, there are few reports about delayed intestinal stenosis after the recovery from NOMI. In this respect, we believe that this case is worth reporting., Introduction Nonocclusive mesenteric ischemia (NOMI) has been reported to be associated with high mortality. Early diagnosis of NOMI and prompt restoration of the intestinal blood flow is necessary in order to achieve a favorable outcome. Presentation of case We present the case of a patient who developed NOMI after autologous blood collection and was treated by selective infusion of the superior mesenteric artery with papaverine, intestinal decompression using a long intestinal tube, the administration of antibiotics, and fluid replacement. Although this non-surgical management was successful, 8 weeks after the ischemic event, segmental bowel resection was necessary because of repeated intestinal obstruction caused by bowel stricture. Discussion Autologous blood collection might be a risk factor of NOMI. In addition, the possibility of delayed intestinal stenosis remains, even if bowel necrosis and surgical resection were avoided with non-surgical management including vasodilator therapy. Conclusion Rapid diagnosis and intervention are essential to minimize intestinal ischemia.

Patients with severe hypophosphatasia (HPP) develop osteogenic impairment with extremely low alkaline phosphatase (ALP) activity, resulting in a fatal course during infancy. Mesenchymal stem cells (MSCs) differentiate into various mesenchymal lineages, including bone and cartilage. The efficacy of allogeneic hematopoietic stem cell transplantation for congenital skeletal and storage disorders is limited, and therefore we focused on MSCs for the treatment of HPP. To determine the effect of MSCs on osteogenesis, we performed multiple infusions of ex vivo expanded allogeneic MSCs for two patients with severe HPP who had undergone bone marrow transplantation (BMT) from asymptomatic relatives harboring the heterozygous mutation. There were improvements in not only bone mineralization but also muscle mass, respiratory function, and mental development, resulting in the patients being alive at the age of 3. After the infusion of MSCs, chimerism analysis of the mesenchymal cell fraction isolated from bone marrow in the patients demonstrated that donor-derived DNA sequences existed. Adverse events of BMT were tolerated, whereas those of MSC infusion did not occur. However, restoration of ALP activity was limited, and normal bony architecture could not be achieved. Our data suggest that multiple MSC infusions, following BMT, were effective and brought about clinical benefits for patients with lethal HPP. Allogeneic MSC-based therapy would be useful for patients with other congenital bone diseases and tissue disorders if the curative strategy to restore clinically normal features, including bony architecture, can be established.

Recent research suggests that raccoons (Procyon lotor) can transmit several important pathogens affecting humans, including protozoans. In Japan, the number of wild raccoons has increased since they were first introduced more than 50 years ago. Here, we report the first survey of Cryptosporidium infection using fecal swabs of raccoons captured in Osaka, Japan. Of 116 raccoons examined by PCR targeting of the Cryptosporidium 18S rRNA gene, 7 (6.03%; 2 adults and 5 young animals) were positive, and the isolates were identified as Cryptosporidium skunk genotype (subtype XVIa) and C. parvum based on sequence and phylogenetic analyses. Both species and the genotype are zoonotic; thus, our results suggest that raccoons could transmit Cryptosporidium infections to humans in Japan.

Aim The Japan Diabetes Society and The Japan Geriatric Society made a joint committee and published a new glycemic target in May 2016. Because reports on the state of glycemic control in elderly diabetic patients are insufficient, we investigated the state of glycemic control in this population before the new glycemic target was established. Methods We enrolled patients older than 65 years of age who had been prescribed antidiabetic drugs and hospitalized in the geriatric department of Nagoya University Hospital from April 1, 2015, to March 31, 2016. We investigated the participants' HbA1c, prescription of antidiabetic drugs carrying risks of severe hypoglycemia (risk drugs) at hospitalization, cognitive function, basic activities of daily living, and instrumental activities of daily living. Results A total of 63 patients were enrolled. Thirty-five patients were male, the mean age was 83.1±5.9 years old, and the average HbA1c was 7.6%±1.5%. The numbers of patients assigned to categories I, II, and III were 10, 12, and 41, respectively. For prescription of risk drugs, 6 participants were assigned to category I, 8 to category II, and 22 to category III. Prescription of risk drugs was associated with the HbA1c. Approximately one third of the patients using risk drugs had a lower HbA1c than the target value. Conclusions Risk drugs was not significantly associated with the established categories or age. Many of the patients who were prescribed risk drugs had a lower HbA1c than the target values.

Background: Although right parasternal approach (RPA) decreases the incidence of mediastinal infection, this approach is associated with lung hernia and flail chest. Our RPA employs thoracotomy with bending rib cartilages and wound closure performed by repositioning the ribs with underlying sheet reinforcement. Methods: We evaluated 16 patients who underwent aortic valve replacement via the RPA from January 2010 to August 2013. We compared outcomes of 15 male patients had the RPA with 30 male patients had full median sternotomy. Results: One patient with a history of radical breast cancer treatment underwent RPA with concomitant right coronary artery bypass grafting. No hospital deaths occurred. Four patients developed hospital-associated morbidity (re-exploration for bleeding, prolonged ventilation, cardiac tamponade, and perioperative myocardial infarction). There were no conversions to full median sternotomy, mediastinal infections, and lung hernias. Preoperative computed tomography showed that the distance from the right sternal border to the aortic root was significantly associated with operation times. With RPA, there was no significant difference in outcomes, despite significantly longer operation times compared with full median sternotomy. Conclusion: Our RPA provides satisfactory outcomes without lung hernia, especially in patients unsuitable for sternotomy. Preoperative computed tomography is useful for identifying appropriate candidates for the RPA.

Cardiovascular manifestations determine the prognosis and survival of patients with Marfan syndrome (MFS). We assessed the early and mid-term outcomes of cardiovascular surgery for this patient population.We conducted a retrospective evaluation of patients with MFS who underwent surgery in our department. The endpoint was the requirement for a second cardiovascular surgery, and death from any cause.From February 2002 to March 2013, we performed 24 surgeries on 13 patients with MFS. At the time of initial surgery, the patients' aged ranged from 19 to 64 years of age (mean 34.7 ± 11.3 years) and included 5 men (38.5 %). Five of 13 initial surgeries (38.5 %) were emergencies. Seven patients (53.8 %) presented with aortic dissection at initial surgery, and one of the remaining six patients suffered from new aortic dissection during follow-up. Eight patients (61.5 %) underwent a second surgery, and a second surgery was not required for 100, 75.0, and 53.6 % of patients at 1, 3, and 5 years, respectively. The mean time interval between the first and the second surgeries was 52.8 ± 28.2 months. No patient died while hospitalized, and three subsequently died from unknown causes. Survival rates were 100, 90.9 and 64.9 % at 3, 5, and 7 years, respectively.Although we obtained satisfactory early outcomes for patients with MFS, there were three late deaths from unknown causes. Therefore, we recommend that patients with MFS should remain under close surveillance.

Patients with total circumferential mitral annular calcification (MAC) extending into the intervalvular fibrous body and aortic annulus have a high risk of cardiac surgery, which remains a technical challenge for surgeons. Our technique for MAC is characterized as simple supra-mitral annular prosthesis insertion after minimum debridement of calcification ("half-and-half technique"). To date, our technique has been applied in only simple MAC cases and has good results. Herein, we report successful two cases of total circumferential MAC, extending into the intervalvular fibrous body and aortic annulus that were treated by a simple double valve replacement with application of our "half-and-half technique".

A 72-year-old man presented with worsening dyspnea on effort. He underwent right pneumonectomy 40 years ago, then mitral valve replacement through a right thoracotomy 8 years ago with repeat surgery to repair a periprosthetic valve leak; the mediastinum was displaced to the right, and the heart was rotated counterclockwise. Transthoracic echocardiography showed periprosthetic valve leak recurrence near the left atrial appendage. We repaired the periprosthetic valve leak through a median sternotomy. Transecting the main pulmonary artery allowed us to widely open the cranial-sided left atrium. We obtained good exposure of the mitral valve, and repaired the periprosthetic valve leak using pledgeted sutures and a pericardial patch.

We developed a microfluidic perfusion cell culture chip that provides three different shear stress strengths and a large cell culture area for the analysis of vascular endothelial functions. The microfluidic network was composed of shallow flow-control channels of three different depths and deep cell culture channels. The flow-control channels with high fluidic resistances created shear stress strengths ranging from 1.0 to 10.0 dyn/cm(2) in the cell culture channels. The large surface area of the culture channels enabled cultivation of a large number (approximately 6.0 × 10(5)) of cells. We cultured human umbilical vein endothelial cells (HUVECs) and evaluated the changes in cellular morphology and gene expression in response to applied shear stress. The HUVECs were aligned in the direction of flow when exposed to a shear stress of 10.0 dyn/cm(2). Compared with conditions of no shear stress, endothelial nitric oxide synthase mRNA expression increased by 50% and thrombomodulin mRNA expression increased by 8-fold under a shear stress of 9.5 dyn/cm(2).

The loss of skeletal muscle mass with aging (sarcopenia) reduces involuntary human activity. To elucidate a mechanism for sarcopenia, the relationship between resistance exercise and gene expression in a muscle has been evaluated. In this paper, we report the development of a microfluidic shredding chip to achieve efficient RNA extraction from microtissues of skeletal muscle. The microfluidic shredding chip was fabricated from hard material and was given high-pressure resistance. The harder microtissue of the skeletal muscle was shredded by permeation through a micropillar array with external forces and the chemical effect of a lysis buffer. The amount of total RNA in the samples increased by twofold because of the microfluidic shredding. The working principle of the microfluidic shredding chip can be widely applied for the extraction of nucleic acids from microtissue. © 2016 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.

Human induced pluripotent stem cells (hiPSCs) are a promising cell source for drug screening. For this application, self-renewal or differentiation of the cells is required, and undefined factors in the culture conditions are not desirable. Microfluidic perfusion culture allows the production of small volume cultures with precisely controlled microenvironments, and is applicable to high-throughput cellular environment screening. Here, we developed a microfluidic perfusion culture system for hiPSCs that uses a microchamber array chip under defined extracellular matrix (ECM) and culture medium conditions. By screening various ECMs we determined that fibronectin and laminin are appropriate for microfluidic devices made out of the most popular material, polydimethylsiloxane (PDMS). We found that the growth rate of hiPSCs under pressure-driven perfusion culture conditions was higher than under static culture conditions in the microchamber array. We applied our new system to self-renewal and differentiation cultures of hiPSCs, and immunocytochemical analysis showed that the state of the hiPSCs was successfully controlled. The effects of three antitumor drugs on hiPSCs were comparable between microchamber array and 96-well plates. We believe that our system will be a platform technology for future large-scale screening of fully defined conditions for differentiation cultures on integrated microfluidic devices.

Under osteoconductive conditions, porous calcium phosphate ceramics are known to induce new bone formation within their pores. A critical aspect of the design of porous ceramics is the geometrical features of their pores, with regard to promoting bone formation and mass transfer management in pore networks. However, the pore geometries of common porous ceramics lack clear details. Further, the connections between pores are hard to characterize and thus have not been thoroughly researched. To address these issues, we have developed an original method for fabricating porous ceramics, which we have termed “mosaic-like ceramics fabrication (MLCF).” Using MLCF, pore geometries can be designed and fabricated by each unit, and a network covering all the pores can be fabricated. Furthermore, MLCF can be used to build porous ceramics with custom-made shapes. In this study, we assessed the osteogenic influences of MLCF products (MLPC) composed of hydroxyapatite units on the differentiation of rat bone-marrow-derived mesenchymal stem cells (MSCs) in vitro and in vivo. Two types of commercial porous artificial bone were used as positive controls. MLPC was superior in osteogenic potential, and proved to be a reliable scaffold for bone tissue engineering. Furthermore, this study succeeded in defining the important geometries for osteoconduction. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 101A: 3571–3579, 2013.

Background: Cell therapies are hampered by the difficulty of delivering cells to and retaining them in target tissues long enough to repair or regenerate local tissues. Hypothesis: Magnetic-assisted delivery of magnetically labeled mesenchymal stem cells (m-MSCs) would be rapid, allowing for chondrogenic differentiation and functional joint repair without replacement. Study Design: Controlled laboratory study. Methods: Sixteen mini-pigs aged 6 to 7 months were used. A full-thickness cartilage defect was created in the center of the patella with a cylindrical punch (diameter, 6 mm). At 4 weeks after creation of the cartilage defects, the animals were divided into 3 treatment groups: In the M group, m-MSCs (5 × 106 cells) were injected and accumulated to the cartilage defect using an external magnetic force (1.5 T) for 10 minutes; in the G group, the patella was faced upward, filled with MSCs (5 × 106 cells), and held for 10 minutes; and in the C group, only phosphate-buffered saline was injected. The regenerated cartilage was evaluated in 5 knees in each of the 3 groups by arthroscopic surgery at 6 and 12 weeks and histological and ultrasound evaluation at 12 and 24 weeks. Results: The mean arthroscopic scores at 6 weeks were 10.4 ± 1.10 in the M group, 8.8 ± 0.84 in the G group, and 7.4 ± 0.89 in the C group. There was a statistically significant difference between the M group and the other 2 groups. The mean arthroscopic scores at 12 weeks were 12.8 ± 1.30 (M group), 10.5 ± 1.30 (G group), and 9.5 ± 0.58 (C group), with a statistically significant difference between the M and C groups. The mean histological scores using the Wakitani scoring system at 12 weeks were 2.8 ± 0.96 (M group), 5.4 ± 0.55 (G group), and 6.0 ± 2.20 (C group), and the mean histological scores at 24 weeks were 2.4 ± 1.50 (M group), 3.5 ± 0.56 (G group), and 5.3 ± 1.50 (C group). The mean histological scores at 12 weeks were significantly better in the M group than in the other groups, and the M group maintained a significantly better histological score than did the C group at 24 weeks. Conclusion: The m-MSCs had no adverse effect on chondrogenic differentiation, and m-MSCs delivered by magnetic field application repaired cartilage defects. Clinical Relevance: The clinical application of this novel stem cell delivery system is a potential therapeutic option for treating cartilage defects and may be more applicable throughout the body than traditional methods.

This paper outlines Buddhist-based meditation in terms of its spiritual, psychotherapeutic, physiological and neuroscientific perspectives. In the latter part of this paper, a pilot study is discussed, in which Japanese university students volunteered to practice meditation at home and complete questionnaires. T-tests were performed to compare with the non-meditated control group. Although only a small number in the experimental group completed the study, our analyses demonstrated that students benefited from meditation and showed significant increases in their sense of coherence, self-esteem and purpose in life. Lastly, practical implications of meditation in contemporary Japanese society are discussed.

Xylitol dehydrogenase (XDHA) and L-arabitol dehydrogenase (LADA) are two key enzymes in xylan metabolism catalyzing the oxidation of xylitol to D-xylulose and arabitol to L-xylulose, respectively. In Aspergillus oryzae, XDHA and LADA are encoded by xdhA and ladA. We deleted xdhA and ladA and xdhA-ladA to generate mutants with decreased dehydrogenase activities and increased xylitol production. The mutants were constructed by homologous transformation into A. oryzae P4 (∆pyrG) using pyrG as a selectable marker. The xylitol productivity of the mutants was measured using D-xylose as the sole carbohydrate source. xdhA, ladA, and the double-deletion mutant produced, respectively, 12.4 g xylitol/l with a yield of 0.24 g/g D-xylose, 12.4 g/l with a yield of 0.33 g/g D-xylose, and 8.6 g/l at a yield of 0.26 g/g D-xylose.

Background The Lachman test is clinically considered to be a reliable physical examination for anterior cruciate ligament (ACL) deficiency. However, the test involves subjective judgement of differences in tibial translation and endpoint quality. An auscultation system has been developed to allow assessment of the Lachman test. The knee joint sound during the Lachman test was analyzed using fast Fourier transformation. The purpose of the present study was to quantitatively evaluate knee joint sounds in healthy and ACL-deficient human knees. Methods Sixty healthy volunteers and 24 patients with ACL injury were examined. The Lachman test with joint auscultation was evaluated using a microphone. Knee joint sound during the Lachman test (Lachman sound) was analyzed by fast Fourier transformation. As quantitative indices of the Lachman sound, the peak sound (Lachman peak sound) as the maximum relative amplitude (acoustic pressure) and its frequency were used. Results In healthy volunteers, the mean Lachman peak sound of intact knees was 100.6 Hz in frequency and −45 dB in acoustic pressure. Moreover, a sex difference was found in the frequency of the Lachman peak sound. In patients with ACL injury, the frequency of the Lachman peak sound of the ACL-deficient knees was widely dispersed. In the ACL-deficient knees, the mean Lachman peak sound was 306.8 Hz in frequency and −63.1 dB in acoustic pressure. If the reference range was set at the frequency of the healthy volunteer Lachman peak sound, the sensitivity, specificity, positive predictive value, and negative predictive value were 83.3%, 95.6%, 95.2%, and 85.2%, respectively. Conclusion Knee joint auscultation during the Lachman test was capable of judging ACL deficiency on the basis of objective data. In particular, the frequency of the Lachman peak sound was able to assess ACL condition.

The authors previously created HAp or CaCO3 formed on or in agarose gels (HAp and CaCO3 gels, respectively) as biocompatible and biodegradable bone graft materials. However, these gels have limitations for bone regeneration. Mesenchymal stromal cells (MSCs) have osteogenic potential and are considered useful for bone tissue engineering. The purpose of this study was to clarify the osteogenic abilities of MSCs loaded in HAp or CaCO3 gels (MSC/HAp and MSC/CaCO3 gels, respectively) using a rat cranial defect model compared to HAp and CaCO3 gels alone. HAp, CaCO3, MSC/Hap, and MSC/CaCO3 gels were prepared for in vivo analyses and implanted into full-thickness bone defects created in the rat cranium. All samples were assessed radiologically and histologically at 4 and 8 weeks after implantation. Using microfocus-computed tomography, an increase in bone formation was observed in the MSC-loaded gels compared to the gels alone. In addition, peripheral quantitative computed tomography revealed higher bone mineral contents in the MSC-loaded gels compared to the gels alone. After transmission X-ray diffraction analyses, the degree of apatite c-axis orientation as a bone quality index of newly formed bone in the MSC-loaded gels was close to that of living cranial bone. Histologically, more extensive bone formation was detected in the MSC-loaded gels compared to gels alone. Overall, MSC/HAp and MSC/CaCO3 gels showed equivalent efficacy for bone regeneration. These findings demonstrate that loading of MSCs into the gels strengthened their osteogenic ability and improved the quality of the newly formed bone. As a result, MSC-loaded gels could represent viable therapeutic biomaterials for bone tissue engineering. Copyright © 2012 John Wiley & Sons, Ltd.

Tissue-engineered medical products (TEMPs) should be evaluated before implantation. Therefore, it is indispensable to establish evaluation protocols in regenerative medicine. Whether or not such evaluation protocols are reasonable is generally verified through a 'round robin' test. However, the round robin test for TEMPs intrinsically includes a deficiency, because 'identical' specimens can not be prepared for TEMPs. The aim of the study was to assess the feasibility and limitations of the round robin test for TEMPs by using a prepared evaluation protocol. We adopted tissue-engineered cartilage constructs as delivered specimens and a protocol of measuring sGAG content as an evaluation protocol proposed to ISO TC150/SC7, which is an invasive, but usually applied, method, although non-invasive methods are keenly required in evaluating TEMPs. The results showed that: (a) the coefficient of variation (CV) of the measured sGAG contents in intralaboratory tests was ~5% at most; (b) the CV of sGAG content in the scheme where each participating laboratory measured different constructs was comparable with that in the scheme where each participating laboratory measured one half of a construct along with the organizing laboratory; (c) the CV caused by factors other than the specimen was ~15%, comparable to that in reproducible experiments in biomedical fields. Based on these results, the study concludes that a round robin test for a TEMP could be valuable, under the condition that the delivered TEMPs are sufficiently reproducible so that the CV of the measured values is

Bone marrow mesenchymal stem cells (MSCs) have been used for bone tissue engineering due to their osteogenic differentiation capability, but their application is controversial. To enhance their capability, we prepared biodegradable gelatin sponges incorporating β-tricalcium phosphate ceramics (GT sponge), which has been shown to possess excellent controlled drug-release properties. The GT sponge was used as a carrier for both rat MSCs and bone morphogenetic protein-2 (BMP-2) and osteogenic differentiation was assessed by subcutaneous implantation of four different kinds of implants, i.e. GT-alone, MSC-GT composites, BMP-GT composites and BMP-GT composites supplemented with MSCs (BMP-MSC-GT) in rats. Two weeks after implantation, histological sections showed new bone formation in the peripheral parts of the BMP-GT and in almost the total volume of the BMP-MSC-GT implants. After 4 weeks, histology as well as microCT analyses demonstrated extensive bone formation in BMP-MSC-GT implants. Gene expression and biochemical analyses of both alkaline phosphatase and bone-specific osteocalcin confirmed the histological findings. These results indicate that the combination of MSCs, GT and BMP synergistically enhances osteogenic capability and provides a rational basis for their clinical application in bone reconstruction.

Various mesenchymal stromal cells (MSCs) have been applied to regenerative medicine. MSCs derived from periodontal tissue could also be a useful cell source for alveolar bone regeneration. However, only a few attempts of direct comparisons have been made between MSCs from periodontal tissues and those from other somatic tissues. The purpose of this study was to clarify the osteogenic characteristics of mesenchymal stromal cells derived from bone marrow (BMSCs), adipose tissue (ASCs) and periodontal ligament (PDLSCs). BMSCs, ASCs and PDLSCs were isolated from Fisher 344 rats. After 1 week of primary culture, stromal cells were subjected to cell surface analysis and osteogenic differentiation. The cells were subcultured for 2 weeks with and without osteogenic supplements (OS), followed by biochemical and histological analyses. With regard to cell surface antigens, all MSCs were positive for CD29 and CD90 and negative for CD45. With regard to osteogenic differentiation, BMSCs with OS had the highest ALP activity, calcium uptake and osteocalcin content. Without OS, PDLSCs had the highest levels of these bone differentiation markers. RT-PCR analysis and histological analysis showed similar trends. These results indicate that PDLSCs are an ideal candidate for alveolar bone regeneration.

A 26-year-old man presented chest oppression. He had pectus excavatum associated with Loeys-Dietz syndrome and a history of redo aortic root replacement with the modified Bentall technique using an 8-mm long interposed graft to the left coronary ostium. Coronary angiography revealed severe stenosis of both left coronary ostium and proximal left anterior descending artery, which was supposed to be resulted from thrombosis in the interposed graft. The left coronary system was bypassed through a left thoracotomy, which was suitable in this patient because the pectus excavatum would prevent harvest of the left internal thoracic artery through re-median sternotomy and to avoid potential sternal reentry injury of the heart. Although the left anterior descending artery was easily accessed under off-pump technique, exposure and anastomosis of the circumflex coronary artery was more difficult than expected without cardiopulmonary bypass as the pectus excavatum and adhesion of the heart prevented anterior shift and rotation of the heart.

We report a perfusion culture microchamber array chip for high throughput drug dose response assays. The microchamber array chip was equipped with a serial dilution microfluidic network for generating a logarithmic concentration profile. Cell suspension, drug solution and media were introduced into the liquid reservoir with a micropipette. The dose response assay was carried out by applying the appropriate pressure through the air vent filters. The dose response of a model anti cancer drug, paclitaxel, on HeLa cells were successfully evaluated by the fluorescent image analysis in the microchamber array chip. Also, the accuracy of our microchamber array chip was confirmed to be equivalent to conventional assay plates in dose response assays. The perfusion culture microchamber array chip, with its simple interface and well-designed microfluidic network, will likely become an advantageous platform for future cell-based assay on microchips.

Background It is difficult to quantitatively evaluate adhesive strength between an implant and the neighboring bone using animal experiments, because the degree of fixation of an implant depends on differences between individuals and the clearance between the material and the bone resulting from surgical technique. Methods A system was designed in which rat bone marrow cells were used to quantitatively evaluate the adhesion between titanium alloy plates and bone plates in vitro. Three kinds of surface treatment were used: a sand-blasted surface, a titanium-sprayed surface and a titanium-sprayed surface coated with hydroxyapatite. Bone marrow cells obtained from rat femora were seeded on the titanium alloy plates, and the cells were cultured between the titanium alloy plates and the bone plates sliced from porcine ilium for 2 weeks. After cultivation, adhesive strength was measured using a tensile test, after which DNA amount and Alkaline phosphatase activity were measured. Findings The seeded cells accelerated adhesion of the titanium alloy plate to the bone plate. Adhesive strength of the titanium-sprayed surface was lower than that of the sand-blasted surface because of lower initial contact area, although there was no difference in Alkaline phosphatase activity between two surface treatments. A hydroxyapatite coating enhanced adhesive strength between the titanium alloy palate and the bone plate, as well as enhancing osteogenic differentiation of bone marrow cells. Interpretation It is believed that this novel experimental method can be used to simultaneously evaluate the osteogenic differentiation and the adhesive strength of an implant during in vitro cultivation.

In this paper we report a perfusion culture microchamber array chip with a serial dilution microfluidic network for analyzing drug dose response over a concentration range spanning 6 orders of magnitude, which is required for practical drug discovery applications. The microchamber array chip was equipped with a pressure-driven interface, in which medium and drug solution were added with a micropipet and delivered into the microfluidic network by pneumatic pressure. We demonstrated that the microchamber array chip could be used to estimate the 50% growth inhibitory concentration using the model anticancer drug paclitaxel and the model cancer cell line HeLa. The results obtained by using the microchamber array chip were consistent with those obtained by a conventional assay using microplates. The microchamber array chip, with its simple interface and well-designed microfluidic network, has potential as an efficient platform for high-throughput dose response assays in drug discovery applications.

The expression of four transcription factors (OCT3/4, SOX2, KLF4, and MYC) can reprogram mouse as well as human somatic cells to induced pluripotent stem (iPS) cells. We generated iPS cells from mesenchymal stromal cells (MSCs) derived from human third molars (wisdom teeth) by retroviral transduction of OCT3/4, SOX2, and KLF4 without MYC, which is considered as oncogene. Interestingly, some of the clonally expanded MSCs could be used for iPS cell generation with 30–100-fold higher efficiency when compared with that of other clonally expanded MSCs and human dermal fibroblasts. Global gene expression profiles demonstrated some up-regulated genes regarding DNA repair/histone conformational change in the efficient clones, suggesting that the processes of chromatin remodeling have important roles in the cascade of iPS cells generation. The generated iPS cells resembled human embryonic stem (ES) cells in many aspects, including morphology, ES marker expression, global gene expression, epigenetic states, and the ability to differentiate into the three germ layers in vitro and in vivo. Because human third molars are discarded as clinical waste, our data indicate that clonally expanded MSCs derived from human third molars are a valuable cell source for the generation of iPS cells.

For the assessment of tissue-engineered cartilage, easy and precise evaluation methods are anticipated. We have investigated the use of a fluorescent image analyser and the application of fluorescence intensity measurements to describe and evaluate a tissue-engineered cartilage-like tissue construct. We evaluated the matrix production of a tissue-engineered cartilage-like tissue (TCL) construct in vitro using the image analyser. The autofluorescence intensity of the TCL construct was determined using the image analyser driven by a software program after 1, 2, 3 and 4 weeks of culture. The autofluorescence intensity of the TCL construct gradually increased with increasing culture time. However, there was very little change in the autofluorescence intensity of gels without articular chondrocytes as controls. The relationship between the autofluorescence intensity of the TCL construct and the hydroxyproline content, reflecting the collagen content, was investigated. There was a significant correlation between the two items. Our findings demonstrate the ability of a commercial fluorescent image analyser to evaluate the matrix production of the TCL construct in vitro on the basis of objective data, such as the autofluorescence intensity. Therefore, the image analyser can be used to judge the success or failure of the in vitro culture process of TCL construct without a biopsy. Copyright © 2010 John Wiley & Sons, Ltd.

The instability of enzymatic glucose sensors has prevented the development of a practical artificial pancreas for diabetic patients. We therefore developed an enzyme-free glucose sensor using the gate effect of a molecularly imprinted polymer (MIP). This sensor has the advantages of improved stability and a simplified manufacturing procedure. An adduct of glucose and 4-vinylphenylboronic acid (VPBA) was synthesized by esterification and was then purified. The copolymer of the glucose/VPBA adduct and methylene bisacrylamide was grafted onto an indium tin oxide electrode surface. Glucose was washed out from the copolymer to obtain an MIP layer. Cyclic voltammetry of ferrocyanide in aqueous solution was performed using an MIP-grafted electrode, and the effect of glucose on the anodic current intensity was evaluated. The anodic current intensity was sensitive to the glucose concentration, and the dynamic range (0-900 mg/dl) covered the typical range of diabetic blood glucose levels. The response time of the MIP-grafted electrode to a stepwise change in the glucose concentration was approximately 3-5 min. Thus, we can conclude that, by taking advantage of its gate effect, it is feasible to use an MIP-grafted electrode as a glucose sensor for monitoring blood sugar in diabetic patients.

Raloxifene acts like estrogen in preventing bone loss in postmenopausal women, but it selectively activates biological responses in bone tissue. It has a direct effect on osteoblasts' differentiation and bone formation in bone marrow culture. However, the point at which raloxifene has an effect on bone marrow-derived mesenchymal stem cells (MSCs), regardless of sex difference, is not known. The purpose of this study was to examine the osteogenic effect of raloxifene on MSCs derived from female and male rats and to assess the sex difference of raloxifene with or without osteogenic supplements (OSs) in the regulation of bone formation.Female and male rat bone marrow cells were cultured with or without OSs. In each experimental group, 10-6 M or 10-8 M raloxifene was added. As a control, cells were cultured without raloxifene. Histologically, mineralization was assessed by alizarin red S staining. Biochemically, alkaline phosphatase (ALP) activity, calcium content, and osteocalcin content were assessed.On histological analysis, mineralized nodules were seen on alizarin red S staining in the groups treated with OS. On the biochemical analysis, OS increased ALP activity, calcium content, and osteocalcin content. Among female groups with OSs, 10-6 M raloxifene significantly increased ALP activity, calcium content, and osteocalcin content compared with the controls. Among male groups, raloxifene had negligible effects.10-6 M Raloxifene had no ossification-inducing effect on female MSCs, but it had an ossification-promoting effect; it had no osteogenic effect on male MSCs. Therefore, raloxifene has a sex difference with regard to its osteogenic effect on MSCs. Moreover, combined treatment with raloxifene plus OS has an effect on female MSCs. These results provide a useful insight into the possible influence of raloxifene after MSC transplantation in clinical practice.

Summary Objective The aim of this study was to investigate whether human articular cartilage can be quantitatively evaluated using a spectrocolorimeter. Materials and methods Human articular cartilage specimens were analyzed using a spectrocolorimeter after macroscopic evaluation using the Outerbridge classification. The cartilage characteristics were examined, the L *, a *, b * colorimetric system, the spectral reflectance distribution and the yellow/red spectral reflectance percentage (Y/R SRP). Moreover, the results of the spectrocolorimetric evaluation were compared with the histological score described by Mankin et al. Results There were significant differences among the macroscopic four grades in the L *, a * and Y/R SRP values. The spectral reflectance distribution of grade 1 cartilage exhibited a gradual increase in the spectral reflectance ratio as the wavelength increased. The spectral reflectance curves of grades 2 to 4 cartilage had dips at a wavelength of around 580nm. Across all the measured wavelengths, there were lower reflectance ratios with the progression of cartilage degeneration. Moreover, correlations were observed between the spectrocolorimetric values and Mankin score. A strong relationship existed between Mankin score and he Y/R SRP values. Conclusions The present study is the first to clearly demonstrate the relationship between spectrocolorimetric evaluation and the degeneration of human articular cartilage. The spectrocolorimeter may be a new quantitative evaluation tool for articular cartilage with clinical potential.

Apicoaortic bypass (AAB), or apicoaortic conduit insertion, is a conventional surgical method that has been regaining attention due to the aging population and the increasing number of repeat surgeries. The indication for the procedure has been extended as an alternative for aortic stenosis when the usual sternotomy or aortic clamping is considered to be difficult, e.g., in patients with severe calcification of the ascending aorta (porcelain aorta), or in patients with a patent coronary artery bypass graft located adjacent to the posterior surface of the sternum. Herein, we report our recent anesthetic management of three patients undergoing AAB. Once the apicoaortic conduit is inserted, blood from the left ventricle is ejected via two routes, the narrowed native aortic valve and the apicoaortic conduit. Thus, it is necessary to elucidate any change in blood flow after the withdrawal of the extracorporeal circulation, by using intraoperative transesophageal echocardiography. Furthermore, if a rigid apical connector is not used, anastomosis of the cardiac apex and conduit is conducted under ventricular fibrillation without the infusion of cardioplegic solution; thus, patients are deemed likely to suffer increased myocardial damage. As a rigid apical connector was not used in the three present patients, the administraction of adequate catecholamines was needed for the withdrawal of the extracorporeal circulation. In addition, because those undergoing AAB often have extremely poor cardiac reserve preoperatively owing to the administration of adequate catecholamines was needed for the withdrawal of the extracorporeal circulation. In the three present patients, anesthetic management was successful, and there were no intraoperative or immediate postoperative complications.

We devised a method of sclerotherapy using OK432. The present study comprised a retrospective followup of sclerotherapy outcomes.Eighteen feet in 18 consecutive patients (seven men, 11 women; mean age, 55 years) were treated with sclerotherapy. Previous surgery had been performed one to four times for seven feet. After aspirating the contents of the ganglion cysts, 0.2 to 0.3 ml of suspension of OK432 (lyophilized strep pyogenes) was injected. The volume of fluid aspirated was from 0.2 to 2.5 ml, with an average of 0.9 ml. Mean duration of followup was 3 years 6 months. Clinical evaluation was performed using the AOFAS hallux scale.All ganglion cysts disappeared after sclerotherapy. However, recurrences were observed in eight feet. Mean duration until recurrence was 13 months. Sclerotherapy was again performed for all patients with recurrence. Re-recurrence occurred in three feet. A third sclerotherapy was performed for all three patients. All ganglion cysts had disappeared by final followup. Mean AOFAS score improved from 75 points before therapy to 95 points at followup. Six patients had complications with local redness and pain for 1 to 6 days after injection.Sclerotherapy using OK432 is a minimally invasive treatment of symptomatic ganglion cysts of the hallux. Recurrence is common but complications are infrequent and self-limited.

Background The modified Bentall technique, which was reported by Svensson in 1992, is an aortic root composite valve graft replacement involving reimplantation of the left coronary ostium with a long interposed graft wrapping behind the composite graft. The technique is technically advantageous, particularly for complicated or redo aortic roots. To justify the technique, the midterm outcome needs to be evaluated. Methods Since 1992, 40 patients (4 with Marfan syndrome) underwent the modified Bentall technique (Svensson's modification). The mean age was 54.7 ± 13.6 years, and 32 patients (80.0%) were male. All hospital survivors have been consecutively followed with annual echocardiographic evaluations. Furthermore, in 2007, multislice computed tomography was performed at 4.7 ± 3.5 years (maximum, 14.9 years) postoperatively in 30 patients who had preserved renal function. Results No patients have experienced any complications regarding the technique at the follow-up of 5.7 ± 4.0 years (maximum, 14.9 years), although there were 2 hospital deaths of emergency cases and 5 late deaths owing to noncardiac causes. In 35 patients (92.1% of hospital survivors), no structural complications were detected by multislice computed tomographies of the 30 patients or coronary angiograms of the remaining 5 patients. The consecutive echocardiographic follow-ups showed well-preserved left ventricular function with the most recent ejection fraction being 0.581 ± 0.078. Conclusions This Svensson's modification technique was associated with favorable midterm outcomes by multislice computed tomography and consecutive echocardiographic evaluations, indicating long-lasting advantages as well as technical benefits. Thus, the technique can be considered as a helpful and justifiable alternative method.

Background: A proximal spherical metatarsal osteotomy was devised to correct not only varus deviation of the first metatarsal, but also dorsiflexion. We expected to increase the medial longitudinal arch by adding plantar flexion at the osteotomy site. To investigate the limitations of this procedure for feet with severe hallux valgus, a followup study was performed on patients with preoperative hallux valgus angles greater than or equal to 40 degrees. Materials and Methods: Forty-eight feet in 37 patients (10 male, 27 female) (60 years; range, 20 to 84 years) were investigated. Mean followup was 4 years and 1 month, ranging from 2 to 8 years. The spherical osteotomy was performed using a curved chisel. A distal soft tissue procedure was done at the same time. Twenty feet received combined operations for their combined deformities. Results: While 81% of patients were satisfied with the results, 50% of those with preoperative hallux valgus angles greater than or equal to 50 degrees had postoperative hallux valgus angle greater than or equal to 20 degrees. In these patients, correction of metatarsus primus varus was good, but correction of valgus deviation of the hallux was fair. Mean correction toward plantar flexion was 1.5 degrees and no increase in arch height was achieved. Conclusion: The proximal spherical osteotomy could consistently achieve satisfactory results for the patients with hallux valgus angles less than 50 degrees. However, the corrections were worse in feet with more severe deformities. Furthermore, plantarflexion at the osteotomy site was offset by displacement at the first tarsometatarsal joint. If plantarflexion is indispensable, arthrodesis of the tarsometatarsal joint is recommended. Level of Evidence: IV, Retrospective Case Series