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Akira Ito

Publications and source records attributed to Akira Ito.

At least 19 recordsLinked to original sources
PubMed2007

Construction of multi-layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force.

Heart tissue engineering requires construction of three-dimensional (3-D) tissues composed of cardiomyocytes (CMs) that are tightly connected to each other. The aim of this study was to construct "scaffold-less" multi-layered 3-D CM sheets using magnetic force-based tissue engineering (Mag-TE) and to evaluate the cell-to-cell functional connections within the CM sheets. Original magnetite cationic liposomes (MCLs) with a positive surface charge (which facilitate adsorption to the target cell surface) were taken up by CMs that were isolated from 2-day-old Wistar rats. When MCLs were added to the medium of CMs at magnetite concentrations of 25, 50, and 100 pg per cell, subsequent measurements showed that 7.2, 13.2, and 27.3 pg of magnetite were taken up per cell, respectively, after 4 h incubation at 37 degrees C. Further, no toxicity was observed after a 24 h incubation period. Using magnetically labeled CMs (magnetite concentration, 100 pg/cell), multi-layered CM sheets were constructed. Immunofluorescent staining of connexin43 demonstrated the presence of gap junctions within the CM sheets that were constructed by Mag-TE. Moreover, electrical connections within the CM sheets constructed by Mag-TE were confirmed using extracellular potential mapping. These results indicate that Mag-TE is a viable methodology for heart tissue engineering.

Animals↗
PubMed2006

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins.

We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for the production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.

Arabidopsis↗
PubMed2006

Complete regression of experimental prostate cancer in nude mice by repeated hyperthermia using magnetite cationic liposomes and a newly developed solenoid containing a ferrite core.

BACKGROUND: Magnetite cationic liposomes (MCLs) can be used to induce hyperthermia because they generate heat in an alternating magnetic field (AMF). This study aimed at developing more practical method for MCL hyperthermia examining the effect of MCL-induced hyperthermia on human prostate cancer in vivo. MATERIALS AND METHODS: A newly developed AMF generator incorporating a solenoid with a ferrite core (FC) was used. Human prostate cancer cells (PC-3 and LNCap) were injected subcutaneously into nude mice. MCLs were injected into tumor nodule and the mice were exposed into AMF three times at 24-hr intervals (repeated hyperthermia; RH) until complete tumor regression was observed. RESULTS: Irradiation with an AMF generated by newly developed device can adequately increase the temperature of tumor tissue. Frequent RH resulted in complete tumor regression in all nude mice. CONCLUSION: RH using MCLs may be a promising new therapy for hormone-refractory human prostate cancer in the future.

Animals↗
PubMed2006

Cluster analysis and gene expression profiles: a cDNA microarray system-based comparison between human dental pulp stem cells (hDPSCs) and human mesenchymal stem cells (hMSCs) for tissue engineering cell therapy.

We investigated gene expression patterns and functional classifications regarding the clusters of human dental pulp stem cells (hDPSCs) and human mesenchymal stem cells (hMSCs)--which possess a multipotent ability--because little is known about the precise moleculobiological clues by which these cells activate their differentiating ability or functionality to eventually form dentin and bone, respectively. We first verified the expressions of the alkaline phosphatase (ALP) gene, dentin matrix protein 1 (DMP-1), and dentinsialophosphoprotein (DSPP) by real-time reverse-transcriptase polymerase chain reaction (RT-PCR) and consequently discovered the high expressions of these genes. Total RNA was also followed by hybridization with a human microarray system consisting of 12,814 genes. Analyses of gene expression patterns indicated several genes which encode extracellular matrix components, cell adhesion molecules, growth factors, and transcription regulators. Functional and clustering analyses of differences in gene expression levels revealed cell signaling, cell communication, or cell metabolism. In the future, information on the gene expression patterns of hDPSCs and hMSCs might be useful in determining the detailed functional roles of the relevant genes and applicable to stem cell therapies, and these cells could also be used as multipotent cell sources for gene technology and tissue engineering technology.

Adult↗
PubMed2006

Echinococcus multilocularis: developmental stage-specific expression of Antigen B 8-kDa-subunits.

Antigen B (AgB) initially found in hydatid cyst fluid of Echinococcus granulosus is a polymeric lipoprotein of 160 kDa, and is an aggregate of several different but homologous small proteins with approximately 8 kDa. Four genes encoding these 8-kDa-subunits have been identified from E. granulosus. In this study we isolated five genes encoding 8-kDa-subunits of AgB from Echinococcus multilocularis. Sequence comparison of isolated cDNA clones demonstrated that one of these five clones was completely identical to EmAgB8/1 which had been isolated previously by our group, and three of them were 94.5, 90.8, and 91.9% homologous to E. granulosus antigen B 8-kDa subunit genes, EgAgB8/2, EgAgB8/3, and EgAgB8/4, respectively. The remaining clone shared 51-58% homology with the nucleotide sequences of AgB genes. Gene-specific RT-PCR and Western blot analyses revealed that these genes were expressed in a developmentally regulated manner in E. multilocularis vesicles, protoscoleces, and immature adult worms. Possible functions of different expression manners are also discussed.

Amino Acid Sequence↗
PubMed2006

A case of intramuscular cysticercosis diagnosed definitively by mitochondrial DNA analysis of extremely calcified cysts.

A case of obsolete intramuscular cysticercosis diagnosed definitively by mitochondrial DNA analysis of extremely calcified cysts was reported. X-ray and computed tomography findings highly suggested cysticercosis due to Taenia solium; however, no direct evidence of cysticercosis was obtained through serological or histopathological examinations. Mitochondrial DNA analysis of a histopathological specimen confirmed the causative agent to be the Asian genotype of T. solium.

Aged, 80 and over↗
PubMed2006

Enhanced cell-seeding into 3D porous scaffolds by use of magnetite nanoparticles.

To engineer functional tissues, a large number of cells must be successfully seeded into scaffolds. We previously proposed a methodology for tissue engineering using magnetite nanoparticles and magnetic force, which we termed "Mag-TE." In the present study, we applied the Mag-TE technique to a cell seeding process and have termed the technique "Mag-seeding." The cell-seeding efficiency of NIH/3T3 fibroblasts (FBs) by Mag-seeding was investigated using six types of commercially available scaffolds (5 collagen sponges and 1 D,D-L,L polylactic acid sponge) having various pore sizes. FBs were magnetically labeled with our original magnetite cationic liposomes (MCLs), which have a positive surface charge, to improve adsorption onto the cell surface. FBs labeled with MCLs were seeded onto a scaffold, and a magnet (4 kG) was placed under the scaffold. Mag-seeding facilitated successful cell seeding into the deep internal space of the scaffolds. Cell-seeding efficiency increased significantly in all scaffolds when compared to those without magnetic force. Moreover, when a high-intensity magnet (10 kG) was used, cell-seeding efficiency was significantly enhanced. These results suggest that Mag-seeding is a promising approach for tissue engineering.

Animals↗
PubMed2006

Extracorporeal cardiac shock wave therapy ameliorates myocardial ischemia in patients with severe coronary artery disease.

OBJECTIVE: Prognosis of severe coronary artery disease with no indication of percutaneous coronary intervention or coronary artery bypass grafting remains poor. We have recently demonstrated that shock wave therapy effectively induces neovascularization and improves myocardial ischemia in a porcine model in vivo. METHODS: With permission from the Ethical Committee of our Institute, we treated nine patients with end-stage coronary artery disease with no indication of percutaneous coronary intervention or coronary artery bypass grafting (55-82 years old, five men and four women) with our cardiac shock wave therapy (200 shots/spot at 0.09 mJ/mm for 20-40 spots, 3 times a week/series). We followed-up the patients at 1, 3, 6, and 12 months after the therapy to examine the amelioration of myocardial ischemia. When needed, shock wave therapy was performed up to three series at 0, and 1, 3 or 6 months. RESULTS: The cardiac shock wave therapy improved symptoms (Canadian Cardiovascular Society functional class score, from 2.7+/-0.2 to 1.8+/-0.2, P<0.01) and reduced nitroglycerin use (from 5.4+/-2.5 to 0.3+/-0.3/week, P<0.05). The treatment also improved myocardial perfusion as assessed by dipyridamole stress thallium scintigraphy (severity score, 25.2+/-7.2% improvement, P<0.05; extent score, 23.3+/-9.0% improvement, P=0.10; washout rate, 20+/-3 to 34+/-3, P<0.05). Myocardial perfusion was improved only in the ischemic area treated with the therapy. These beneficial effects persisted for 12 months. No procedural complications or adverse effects were noted. CONCLUSION: These results indicate that our extracorporeal cardiac shock wave therapy is an effective and non-invasive treatment for end-stage coronary artery disease, although further careful evaluation is needed.

Aged↗
PubMed2006

Assessment of real-time PCR for diagnosis of Mycoplasma pneumoniae pneumonia in pediatric patients.

We developed a real-time PCR to detect Mycoplasma pneumoniae with a primer set designed for the 16S rRNA gene. Clinical samples (n=937) were collected from children with community-acquired pneumonia between April 2002 and March 2004 at 12 Japanese medical institutions. Sensitivity of real-time PCR was calculated as 10 colony-forming units per reaction tube using a pMP01 plasmid carrying a 225-bp target DNA fragment of the 16S rRNA gene in M. pneumoniae M129, a standard strain. Results, obtained within 2 h, were compared with those of conventional culture and serologic methods. Of all cases tested, 151 (16.4%) and 129 (13.8%) were positive for M. pneumoniae by real-time PCR and by culture, respectively. Among the 151 cases, almost all of those tested serologically by passive agglutination showed a rise in M. pneumoniae antibody titre between acute and convalescent sera. We conclude that this real-time PCR can identify M. pneumoniae rapidly and fulfills the need for rapid identification, high sensitivity, and high specificity.

Adolescent↗
PubMed2006

Magnetic resonance imaging of alveolar echinococcosis experimentally induced in the rat lung.

Pulmonary alveolar echinococcosis (AE) caused by the metacestode of Echinococcus multilocularis is a lethal zoonosis and is a lesion secondarily induced by hematogenous dissemination from hepatic AE lesions. In the present study, a hematogenous pulmonary AE model was experimentally induced in rats by the injection of echinococcal larval tissue homogenate to the tail vein, and then the pathological and diagnostic aspects of pulmonary AE were examined by magnetic resonance imaging (MRI). Histological primary, mature and degenerated AE lesions were observed 5, 18 and 50 weeks after injection, respectively. These lesions were discriminated as signal-void, hypointense and hyperintense regions in T1-weighted MRI (T1WI), respectively. The change in signal intensity in T1WI might reflect the content of proteinaceous fluid as a result of AE cyst degeneration. Western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis provided evidence for AE infection in the early stage. T1WI in combination with Western blot analysis could possibility become definitive and early signs of hematogenous pulmonary AE infection.

Animals↗
PubMed2006

Unique family clustering of human echinococcosis cases in a chinese community.

We have identified a significant focus and unusual clustering of human cases of cystic echinococcosis (CE) and alveolar echinococcosis (AE) in the village of Nanwan, Xiji County, Ningxia Hui Autonomous Region, in one of the most highly endemic areas for both diseases in China. The village, a Chinese Hui Islamic community, is composed of 167 members of four extended families. A total of 28 people died (12 of echinococcosis) since the village was first settled in the 1950s. Despite similar life patterns, the number of AE and CE cases occurring in each family was different. Overall, the prevalences of AE and CE were 9% (20 cases) and 5.9% (13 cases), with a combined prevalence of 14.9%. In contrast to CE, a comparison of the prevalence of AE indicated significant differences between the four family clusters. Although suggestive that host genotype might play a role in susceptibility to AE, this hypothesis requires further investigation.

Adolescent↗
PubMed2006

Dual infection of animal hosts with different Echinococcus species in the eastern Qinghai-Tibet plateau region of China.

The eastern Qinghai-Tibet plateau of China is a highly endemic region of echinococcosis where Echinococcus granulosus sensu stricto (sheep strain), Echinococcus multilocularis, and Echinococcus shiquicus are distributed sympatrically. We developed a polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method for the identification of the three species in this region. The PCR-RFLP showed the dual infection of animals with different Echinococcus spp. The first case was a domestic dog concurrently infected with adults of E. granulosus and E. multilocularis. The second case was a plateau pika (Ochotona curzoniae) harboring metacestodes of E. multilocularis and E. shiquicus in the liver. The high susceptibility of some mammalian hosts to the parasites and the high prevalence of the three co-endemic species probably increase the chance of mixed infections in the eastern Tibetan plateau.

Animals↗
PubMed2006

[A case of liver cysts with Em18-WB was useful for differential diagnosis].

A 72-year-old woman with multilocular liver cysts was admitted. This lesions seemed to be an alveolar hydatid disease. Two methods of Western blotting were used for serologic diagnosis. One method recognizes antigens of crude extracts of Echinococcus multilocularis (EM). The other method recognizes a purified antigen (Em18) of EM. Her serum only reacted with the former method and never reacted with Em18 antigen. Eighteen months after first admission, she had an operation. The resected specimen was diagnosed with simple cysts. Western blotting using Em18 antigen could greatly facilitate the differential diagnosis of simple liver cyst and alveolar hydatid disease.

Aged↗
PubMed2005

Challenges for control of taeniasis/cysticercosis in Indonesia.

Taeniasis/cysticercosis has been reported from several provinces of Indonesia: Papua (=former Irian Jaya), Bali, North Sumatra, East Nusa Tenggara, South East Sulawesi, Lampung, North Sulawesi, Jakarta, West Kalimantan, and East Java. The highest level of endemicity of taeniasis/cysticercosis has been found in Papua. Recent surveys in Jayawijaya District of Papua in 2000 and 2001 showed that 5 of 58 local people (8.6%) harbored the adult tapeworm, Taenia solium, whereas 44 of 96 people (45.8%), 50 of 71 pigs (70.4%), and 7 of 64 local dogs (10.9%) were seropositive for T. solium cysticercosis. Current surveys in Bali and Samosir District, North Sumatra during 2002-2005 revealed that Taenia saginata taeniasis has increased in incidence whereas T. solium cysticercosis is now rather rare compared to one-two decades ago in Bali. Taenia asiatica taeniasis is still common in Samosir District. Data from other provinces of Indonesia are very limited or unavailable. Control of these diseases is not a priority in the health or veterinary services, neither at central or local government levels. However, limited efforts toward control of the diseases have been implemented such as training of health personnel, community education on disease prevention, and provision of anthelminthics. A working group for control of the disease in Indonesia and an international collaboration have been established among Ministry of Health, Indonesia; University of Indonesia; and Asahikawa Medical College, Asahikawa, Japan since 1996. Future goals include implementation of active case finding (active surveillance) and treatment of tapeworm carriers, sustainable public health education, establishment of a system to check the quality of beef/pork and determine the distribution of infected animals and strengthening of laboratory capacity. Efforts to motivate provinces and districts should be implemented in developing the strategic plan to control of the disease. Given the considerable differences in cultures, religions, levels of education, socio-economic levels, daily habits and behaviour etc., control programmes must be adapted to the local situations.

Animals↗
PubMed2005

Taeniasis/cysticercosis in Papua (Irian Jaya), Indonesia.

Reports showed that an important parasitic zoonotic disease caused by Taenia solium, Taenia saginata and Taenia asiatica is found endemic in several areas of Indonesia including Papua, Bali and North Sumatra. At present it is known that the highest prevalence of taeniasis/cysticercosis in Indonesia, caused by T. solium is among the indigenous communities in Papua (formerly Irian Jaya). In the early 1970s, 8-9% of stool samples from the Enarotali hospital, Paniai District (Irian Jaya) were found positive with Taenia eggs. The samples were from members of the Ekari (Kapauku) ethnic group. Stool samples from the Moni ethnic group, living east of surrounding lakes, were egg negative. Cysticerci of T. solium were discovered in pigs. During the years 1973-1976 cases of burns increased and were ultimately found to be primarily associated with epileptic seizures induced by neurocysticercosis cases. Among 257 cases of burns, 88 cases (62.8%) were suffering from epileptic seizures before or during hospitalization. In the year 1981 T. solium seropositive persons were mostly (16%) found in the endemic Obano village. In 1997 the parasite was discovered in Jayawijaya District, which is located approximately 250 km east of Paniai District. During 1991-1995, a local health center in Assologaima, Jayawijaya District reported 1120 new cases with burns and a further 293 new cases of epileptic seizures among 15,939 inhabitants. The histopathologic appearance and mitochondrial DNA analysis found the cysts to be similar to those of T. solium from other regions of the world. Sensitive and specific serological diagnostic methods were used and improved. Cysticerci were detected in dogs, as well as in pigs. A coproantigen test for detection of adult tapeworms in patients was carried out. Medical treatment with praziquantel for taeniasis and albendazole for cysticercosis (with prednisone and sodium phenytoin treatment in cases with neurocysticercosis) was undertaken. Lifestyle, religion, and socioeconomic aspects are important issues in the perpetuation and enhancing the endemicity of T. solium taeniasis and cysticercosis in Papua, Indonesia.

Animals↗
PubMed2005

Taeniasis and cysticercosis in Bali and North Sumatra, Indonesia.

It has been reported that three human Taenia species are distributed in Indonesia: Taenia solium, Taenia asiatica and Taenia saginata. T. asiatica is well known in North Sumatra, especially on Samosir island in Lake Toba. T. solium and T. saginata are known from Bali. T. solium is most serious public health issue in Papua (former Irian Jaya). In this report, we briefly review the present situation of these three human Taenia species mainly in Bali and North Sumatra. For community based epidemiological survey, we have adopted and applied questionnaire, microscopic observation of eggs, coproantigen tests, coproDNA tests, mitochondrial DNA analysis and morphology for isolated specimens, serology for cysticercosis and taeniasis. For detection of taeniasis cases of both T. saginata and T. asiatica, questionnaire by expert doctors is perfect to detect worm carriers. Corpoantigen test developed for detection of taeniasis of T. solium is also highly reliable to detect taeniasis carriers in the community. Epidemiological data from these two islands 2002-2005 are reviewed with backgrounds of historical culture, religions and customs.

Animals↗
PubMed2005

Recent advances in characterization of Echinococcus antigen B.

Antigen B (AgB) in hydatid cyst fluid of Echinococcus granulosus is a polymeric lipoprotein of 160 kDa and a highly immunogenic major antigen in echinococcal infection. The antigen is comprised of a group of subunit monomers of approximately 8 kDa in molecular size. Recent studies have revealed that the E. granulosus AgB (EgAgB) shows a high degree of genetic variability and the genes encoding the EgAgB 8-kDa subunit monomers that have been identified to date could be grouped into four clades, corresponding to the genes EgAgB8/1, EgAgB8/2, EgAgB8/3 and EgAgB8/4. It has been suggested that the recombinant EgAgB8/2 (rEgAgB8/2) provides better performance in serodiagnosis of human cystic echinococcosis (CE) than does the recombinant EgAgB8/1 (rEgAgB8/1). The EgAgB has been identified as a protease inhibitor with an ability to inhibit recruitment of neutrophils and exploit activation of T helper cells by eliciting a non-protective Th2 cell response, predominantly in patients with progressive CE. Recently it has been revealed that AgB also exists in the cyst fluid of Echinococcus multilocularis. Five different cDNAs encoding the EgAgB homologues have been identified in vesicles, protoscoleces and/or immature adult worms of E. multilocularis and named as EmAgB8/1, EmAgB8/2, EmAgB8/3, EmAgB8/4 and EmAgB8/5. These genes appeared to be expressed in a developmentally regulated manner in the parasite life cycle. This review focuses on recent advances in molecular biological and immunological characterization of AgB from both of E. granulosus and E. multilocularis.

Animals↗