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Biomedical subjects

Yusuke Yamamoto

Publications and source records attributed to Yusuke Yamamoto.

14 recordsLinked to original sources

FGF-4 regulates neural progenitor cell proliferation and neuronal differentiation.

The FGF-4 (fibroblast growth factor 4, known as HST-1) protein is an important mitogen for a variety of cell types. However, only limited information is available concerning tissue distribution and the biological role of FGF-4 in the brain. In situ hybridization analysis revealed localization of mouse Fgf-4 mRNA in the normal postnatal mouse hippocampus, subventricular zone (SVZ), and the rostral migratory stream where new neurons generate, migrate, and become incorporated into the functional circuitry of the brain. We also investigated whether FGF-4 could promote both proliferation and differentiation of the neural progenitor cells by using an in vitro neurosphere assay. The addition of recombinant FGF-4 generated large proliferative spheres that have a multipotent differentiation ability. Furthermore, recombinant FGF-4 significantly promotes neuronal differentiation in attached clonal neurosphere culture. These findings suggest that FGF-4 has an ability to promote neural stem cell proliferation and neuronal differentiation in the postnatal brain.

Animals↗

Bacterial community changes in diesel-oil-contaminated soil microcosms biostimulated with Luria-Bertani medium or bioaugmented with a petroleum-degrading bacterium, Pseudomonas aeruginosa strain WatG.

The microbial community structure of diesel-oil-contaminated soil microcosms biostimulated with Luria-Bertani medium (LB-BS) or bioaugmented with a petroleum-degrading bacterium, Pseudomonas aeruginosa strain WatG (WatG-BA), was investigated by denaturing gradient gel electrophoresis (DGGE) and by monitoring diesel oil degradation. The degradation in WatG-BA (64.0% +/- 4.2%) was higher than that in LB-BS (49.5% +/- 12.0%) during the first two weeks. The microbial community in WatG-BA, which was markedly dominated by strain WatG, was much simpler than that in LB-BS, where hydrocarbon degraders occurred after a lag of 3-7 days after the addition of diesel oil. The clustering profiles of the DGGE banding patterns of the two soil microcosms were only 12% similar. This difference is probably due to antibacterial substances, such as rhamnolipids, secreted by strain WatG.

Actinobacteria↗

Isolation and characterization of a novel thraustochytrid-like microorganism that efficiently produces docosahexaenoic acid.

A thraustochytrid-like microorganism (strain 12B) was isolated from the mangrove area of Okinawa, Japan. On the basis of its ectoplasmic net structure and biflagellate zoospores we determined strain 12B to be a novel member of the phylum Labyrinthulomycota in the kingdom Protoctista. When grown on glucose/seawater at 28 degrees C, it had a lipid content of 58% with docosahexaenoic acid (DHA; 22:6 n-3) at 43% of the total fatty acids. It had a growth rate of 0.38 h(-1). The DHA production rate of 2.8 +/- 0.7 g l(-1) day(-1) is the highest value reported for any microorganism.

Docosahexaenoic Acids↗

[STI571 for gastrointestinal stromal tumors].

Since the effectiveness of STI571 for GIST was reported, therapy for GIST has changed markedly, and the disease has attracted attention. We have treated 19 patients with GIST since 2000 by 19 resections (local resection in 14 patients, total gastrectomy in 2 patients, distal gastrectomy in 2 patients, and hepatectomy in 1 patient), and administered STI571 to 5 patients with unresectable or recurrent GIST. Of these 5 patients, 2, 1, 1, and 1 had PR, SD, PD, and inevaluable disease, respectively, with a response rate of 50%. The disease was controllable in 80% of the patients. All patients had palpebral and crural edema as side effects, which were not severe, suggesting the safety of the drug. It is important to tailor therapy (STI571 or surgery) to the patient.

Aged↗

Efficient delivery of small interfering RNA to bone-metastatic tumors by using atelocollagen in vivo.

Silencing of gene expression by small interfering RNAs (siRNAs) is rapidly becoming a powerful tool for genetic analysis and represents a potential strategy for therapeutic product development. However, there are no reports of systemic delivery for siRNAs toward treatment of bone-metastatic cancer. Accordingly, we report here that i.v. injection of GL3 luciferase siRNA complexed with atelocollagen showed effective reduction of luciferase expression from bone-metastatic prostate tumor cells developed in mouse thorax, jaws, and/or legs. We also show that the siRNA/atelocollagen complex can be efficiently delivered to tumors 24 h after injection and can exist intact at least for 3 days. Furthermore, atelocollagen-mediated systemic administration of siRNAs such as enhancer of zeste homolog 2 and phosphoinositide 3'-hydroxykinase p110-alpha-subunit, which were selected as candidate targets for inhibition of bone metastasis, resulted in an efficient inhibition of metastatic tumor growth in bone tissues. In addition, upregulation of serum IL-12 and IFN-alpha levels was not associated with the in vivo administration of the siRNA/atelocollagen complex. Thus, for treatment of bone metastasis of prostate cancer, an atelocollagen-mediated systemic delivery method could be a reliable and safe approach to the achievement of maximal function of siRNA.

Animals↗

Recapitulation of in vivo gene expression during hepatic differentiation from murine embryonic stem cells.

Hepatic differentiation at the molecular level is poorly understood, mainly because of the lack of a suitable model. Recently, using adherent monoculture conditions, we demonstrated the direct differentiation of hepatocytes from embryonic stem (ES) cells. In this study, we exploited the direct differentiation model to compare the gene expression profiles of ES cell-derived hepatocytes with adult mouse liver using DNA microarray technology. The results showed that the ES cell-derived hepatocyte gene expression pattern is very similar to adult mouse liver. Through further analysis of gene ontology categories for the 232 most radically altered genes, we found that the significant categories related to hepatic function. Furthermore, through the use of small interfering RNA technology in vitro, hepatocyte nuclear factor 3beta/FoxA2 was identified as having an essential role in hepatic differentiation. These results demonstrate that ES cell-derived hepatocytes recapitulate the gene expression profile of adult mouse liver to a significant degree and indicate that our direct induction system progresses via endoderm differentiation. In conclusion, our system closely mimics in vivo hepatic differentiation at the transcriptional level and could, therefore, be useful for studying the molecular basis of hepatocyte differentiation per se.

Animals↗

Giant pleomorphic adenoma of the parotid gland with malignant transformation.

A 72-year-old female is described who was admitted to the hospital with a giant mixed tumor of the left parotid gland that had grown to a weight of 6051 g within 20 years and ultimately underwent resection of the mass. Histopathological examination revealed adenocarcinoma cells with hyperchromatic nuclei and mitotic figures proliferated in pleomorphic adenoma. Many authors have reported giant tumors weighing more than a few kilograms of the parotid glands, and most were benign. The present tumor was the sixth largest among all the parotid tumors, including both benign and malignant, which were reported. Furthermore, our case was the largest one confirmed as malignant recorded in the world literature. The tumor was successfully resected with no marked facial nerve paralysis.

Adenoma, Pleomorphic↗

Long-term maintenance of liver-specific functions in cultured ES cell-derived hepatocytes with hyaluronan sponge.

This study investigated the three-dimensional culture of hepatocytes differentiated from mouse embryonic stem (ES) cells with a porous hyaluronan (HA) sponge support. Hepatocytes were immobilized within the pores of the support. Spheroids could be observed within the support, each containing between 20 and 50 hepatocytes. To examine the liver-specific functions of the hepatocytes in the culture, the levels of albumin secreted into the medium were analyzed. The secretion of albumin was stable over the course of 32 days, longer than that in both conventional monolayer and collagen sponge cultures. To elucidate further the liver-specific functions of hepatocytes embedded in the HA sponge, metabolic activities of the hepatocytes were examined for their ability to eliminate ammonia from culture media and the synthesis of urea nitrogen. While rates of ammonia removal and urea nitrogen synthesis were similar to those in both conventional monolayer and in collagen sponge cultures, these functions were maintained for longer duration in cells embedded in the HA sponge. These results demonstrate that the porous HA sponge is an effective support for the in vitro culture of ES-derived hepatocytes used for both basic and applied studies for cell transplantation.

Animals↗

Atelocollagen-mediated synthetic small interfering RNA delivery for effective gene silencing in vitro and in vivo.

Silencing gene expression by siRNAs is rapidly becoming a powerful tool for the genetic analysis of mammalian cells. However, the rapid degradation of siRNA and the limited duration of its action call for an efficient delivery technology. Accordingly, we describe here that Atelocollagen complexed with siRNA is resistant to nucleases and is efficiently transduced into cells, thereby allowing long-term gene silencing. Site-specific in vivo administration of an anti-luciferase siRNA/Atelocollagen complex reduced luciferase expression in a xenografted tumor. Furthermore, Atelocollagen-mediated transfer of siRNA in vivo showed efficient inhibition of tumor growth in an orthotopic xenograft model of a human non-seminomatous germ cell tumor. Thus, for clinical applications of siRNA, an Atelocollagen-based non-viral delivery method could be a reliable approach to achieve maximal function of siRNA in vivo.

Animals↗

Generation of cloned calves and transgenic chimeric embryos from bovine embryonic stem-like cells.

Bovine embryonic stem-like cells (ES-like cells) appear to maintain a normal diploid karyotype indefinitely during culture in vitro and to express marker proteins that are characteristic of ES cells from mice, namely, alkaline phosphatase (AP), stage-specific embryonic antigen-1 (SSEA-1), STAT-3, and Oct 4. After proliferation of undifferentiated ES-like cells in vitro, some bovine ES-like cells differentiated to neural precursor cells, which were cultured in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), and platelet-derived growth factor (PDGF). In addition, calves were successfully cloned using ES-like cells and the frequency of term pregnancies for blastocysts derived from ES-like cells was higher than those of early pregnancies and maintained pregnancies after nuclear transplantation (NT) with bovine somatic cells. Successful cloning from bovine ES-like cells should allow the introduction into cattle of specific genetic characteristics of biomedical and/or agricultural importance.

Animals↗

Optic disc excavation in the atrophic stage of Leber's hereditary optic neuropathy: comparison with normal tension glaucoma.

BACKGROUND: Abnormal optic disc excavations are reportedly seen in patients with Leber's hereditary optic neuropathy (LHON), a mitochondrial dysfunction disease. We examined the disc morphology in the eyes of patients with LHON at the atrophic stage and compared it to that in eyes with normal-tension glaucoma (NTG). METHODS: We studied 15 LHON patients with the 11778 mutation, 15 patients with NTG, and 25 normal subjects. The optic disc morphology was analyzed by Heidelberg retinal tomography (HRT). Ten parameters of the optic disc obtained by HRT were evaluated, including the diagnostic classification of glaucoma. RESULTS: Six of the nine morphological HRT parameters of the LHON patients, the exceptions being disc area, mean cup depth, and maximum cup depth, differed significantly from those of the normals. NTG patients had a significantly greater mean and maximum cup depth than LHON patients. The HRT glaucoma diagnostic software classified 22 (73%) of the 30 optic discs in LHON patients as glaucomatous. CONCLUSION: The optic discs at the atrophic stage of LHON eyes have glaucoma-like morphological changes. However, the cups were significantly deeper in NTG than LHON. The similarity in the optic disc findings in LHON and NTG suggests that alterations in mitochondrial function may be related to optic disc excavations.

Adolescent↗

Isolation of embryonic stem-like cells from equine blastocysts and their differentiation in vitro.

Embryonic stem (ES) cells are pluripotent cells with the potential capacity to generate any type of cell. We describe here the isolation of pluripotent ES-like cells from equine blastocysts that have been frozen and thawed. Our two lines of ES-like cells (E-1 and E-2) appear to maintain a normal diploid karyotype indefinitely in culture in vitro and to express markers that are characteristic of ES cells from mice, namely, alkaline phosphatase, stage-specific embryonic antigen-1, STAT-3 and Oct 4. After culture of equine ES-like cells in vitro for more than 17 passages, some ES-like cells differentiated to neural precursor cells in the presence of basic fibroblast growth factor (bFGF), epidermal growth factor and platelet-derived growth factor. We also developed a protocol that resulted in the differentiation of ES-like cells in vitro to hematopoietic and endothelial cell lineages in response to bFGF, stem cell factor and oncostatin M. Our observations set the stage for future developments that may allow the use of equine ES-like cells for the treatment of neurological and hematopoietic disorders.

Animals↗