Total syntheses of disulphated glycosphingolipid SB1a and the related monosulphated SM1a

Total syntheses of two natural sulphoglycolipids, disulphated glycosphingolipid SB1a and the structurally related monosulphated SM1a, are described. They have common glycan sequences and ceramide moieties and are associated with human epithelial carcinomas. The syntheses featured efficient glycan assembly and the glucosyl ceramide cassette as a versatile building block. The binding of the synthetic sulphoglycolipids by the carcinoma-specific monoclonal antibody AE3 was investigated using carbohydrate microarray technology

In Vitro and In Vivo Evaluation of Starfish Bone-Derived -Tricalcium Phosphate as a Bone Substitute Material

We evaluated starfish-derived -tricalcium phosphate (Sf-TCP) obtained by phosphatization of starfish-bone-derived porous calcium carbonate as a potential bone substitute material. The Sf-TCP had a communicating pore structure with a pore size of approximately 10 m. Although the porosity of Sf-TCP was similar to that of Cerasorb M (CM)a commercially available -TCP bone fillerthe specific surface area was roughly three times larger than that of CM. Observation by scanning electron microscopy showed that pores communicated to the inside of the Sf-TCP. Cell growth tests showed that Sf-TCP improved cell proliferation compared with CM. Cells grown on Sf-TCP showed stretched filopodia and adhered; cells migrated both to the surface and into pores. In vivo, vigorous tissue invasion into pores was observed in Sf-TCP, and more fibrous tissue was observed for Sf-TCP than CM. Moreover, capillary formation into pores was observed for Sf-TCP. Thus, Sf-TCP showed excellent biocompatibility in vitro and more vigorous bone formation in vivo, indicating the possible applications of this material as a bone substitute. In addition, our findings suggested that mimicking the microstructure derived from whole organisms may facilitate the development of superior artificial bone.ArticleMATERIALS. 12(11):1881 (2019)journal articl

Evaluation of MC3T3-E1 Cell Osteogenesis in Different Cell Culture Media

Many biomaterials have been evaluated using cultured cells. In particular, osteoblast-like cells are often used to evaluate the osteocompatibility, hard-tissue-regeneration, osteoconductive, and osteoinductive characteristics of biomaterials. However, the evaluation of biomaterial osteogenesis-inducing capacity using osteoblast-like cells is not standardized; instead, it is performed under laboratory-specific culture conditions with different culture media. However, the effect of different media conditions on bone formation has not been investigated. Here, we aimed to evaluate the osteogenesis of MC3T3-E1 cells, one of the most commonly used osteoblast-like cell lines for osteogenesis evaluation, and assayed cell proliferation, alkaline phosphatase activity, expression of osteoblast markers, and calcification under varying culture media conditions. Furthermore, the various media conditions were tested in uncoated plates and plates coated with collagen type I and poly-L-lysine, highly biocompatible molecules commonly used as pseudobiomaterials. We found that the type of base medium, the presence or absence of vitamin C, and the freshness of the medium may affect biomaterial regeneration. We posit that an in vitro model that recapitulates in vivo bone formation should be established before evaluating biomaterials.ArticleInternational Journal of Molecular Sciences. 22(14):7752 (2021)journal articl

Cellular Responses of Human Lymphatic Endothelial Cells to Carbon Nanomaterials

One of the greatest challenges to overcome in the pursuit of the medical application of carbon nanomaterials (CNMs) is safety. Particularly, when considering the use of CNMs in drug delivery systems (DDSs), evaluation of safety at the accumulation site is an essential step. In this study, we evaluated the toxicity of carbon nanohorns (CNHs), which are potential DDSs, using human lymph node endothelial cells that have been reported to accumulate CNMs, as a comparison to fibrous, multi-walled carbon nanotubes (MWCNTs) and particulate carbon black (CB). The effect of different surface characteristics was also evaluated using two types of CNHs (untreated and oxidized). In the fibrous MWCNT, cell growth suppression, as well as expression of inflammatory cytokine genes was observed, as in previous reports. In contrast, no significant toxicity was observed for particulate CB and CNHs, which was different from the report of CB cytotoxicity in vascular endothelial cells. These results show that (1) lymph endothelial cells need to be tested separately from other endothelial cells for safety evaluation of nanomaterials, and (2) the potential of CNHs as DDSs.ArticleNANOMATERIALS. 10(7):1374 (2020)journal articl

南部黒瀬川帯寒谷層と坂州不整合

The Sakashu Unconformity, a representative unconformity between Late Permian accretionary comprex and Late Triassic shallow-marine sediments in the South Kurosegawa Terrane, has investigated in stratigraphy, sedimentary facies, allochthonous and autochthonous faunas with the view point of post-accretional environmental changes. The accretion mélange of the Hisone Group, basement of the unconformity, yields Late Permian radiolarians. The lower member of the Upper Triassic Sabudani Formation is associated with two types of debris flows. One contains blocks of pelagic chert, carbonates, and greestones. The other is composed of mylonitized granitic breccias and fragments. The latter probably derived from the Mitaki Granites (400+ Ma) of the Kurosegawa Tectonic Zone that form the substratum of the Silurian limestone. The basal middle member of the Sabudani Formation contains huge blocks of weathered limestone/chert that yields late Early Permian conodont fauna (M. bisseli - S. whitei Zone). The hummocky cross-stratification in the middle member of the Sabudani Formation indicates a lower off-shore facies. Existence of the Permian conodont-bearing huge eroded limestone/chert blocks, and early Late Permian radiolarians in chert pebbles of the hummocky cross-stratified lag sediments suggests probable derivation of the conglomerates from the blocks in the Permian accretionary complexes. The Triassic mollusk fauna of the middle and upper members of the Sabudani Formation is correlative with the Carnian (-Norian) Kochigatani bivalve-fauna in the clastic facies of the Kurosegawa Terrane (Outer Zone of SW Japan) as well as with the Mine and Nabae groups, Primorye, Zabaikal and NE Siberia. From a paleobiogeographic point of view, the bivalve-fauna of the clastic sand- and marlstone Kochigatani facies is regarded to differ from that of the carbonate Tethyan facies in the South Chichibu Jurassic Accretionary Terrane

Marbles from Tokushima Prefecture, used in the National Diet Building : Part 1: locality and geological framework

The quarries of marbles for the construction of the National Diet Building (Nagata-cho, Chiyodaku, Tokyo) were identified in Tokushima Prefecture. The geological setting and microfossil age of the marbles were studied. Seven brands, "Akebono", "Kito-ishi ", "Hototogisu", "Kamo-sarasa", "Awayuki ", "Kotajima" and "Shin-awayuki" were used in the House of Councilors and the Central part of the building. The "Akebono", used for the wall along the Councilor's Second Stairway, is Silurian limestone from the Hiakari-yama lenticular body of the Kurosegawa Tectonic Zone in the Koyama-daira area (Kisawa Village). The "Kito-ishi" from the Misago-yama area (Kisawa Village), used for the fireplace in the reception room of the Vice-president of the House of Councilors, is the limestone and greenstone breccia intercalated in the Upper Carboniferous alternation of chert and pelagic limestone in the Permian accretion terrene (Masaki Belt of the Kurosegawa Terrane). The "Hototogisu", used for the well-ornamented entrance gate of the Emperor's room "Gokyujo", is the Upper Triassic dolimitic limestone in the Late Jurassic-earliest Cretaceous accretion mélange of the South Chichibu Terrane (Tsunomine Subbelt) from the Arida area (Asebi-cho, Anan City). The "Kamo-sarasa", used for the inside wall of the central entrance of the building, is the Upper Triassic limestone with some green reticulation in Late Jurassic-earliest Cretaceous accretion mélange of the South Chichibu Terrane (Tsunomine Subbelt) from the Oji area (Kuwano-cho, Anan City). The "Awayuki", used for the lower part of the wall along the square in front of the Emperor's room, and the lower part of the wall around the assembly hall of the House of Councilors, presumed to be a kind of "Kamo-sarasa", is proved to be the Upper Triassic Megalodon limestone with hexacoral-bearing dark limestone blocks in the accretion mélange of the South Chichibu Terrane (Tsunomine Subbelt) from the Kurogo area (Kamo-cho, Anan City). The "Kotajima", used for the lower part of the wall along the passage of the second floor, and the wall along the "Kisha Kaidan" stairway, is proved to be the Upper Triassic limestone with reticulation of black cleavages in Late Jurassic-earliest Cretaceous accretion mélange of the South Chichibu Terrane (Tsunomine Subbelt) from the Higashibun area (Tsunomine-cho, Anan City). The "Shin-awayuki", used for the mosaic on the floor of the Central Hall and the lower part of the wall along the passage of the southern central part of the first floor, is the Upper Carboniferous limestone in Jurassic accretion mélange of the South Chichibu Belt (Hosono Subbelt) from the Izeki area (Takarada-cho, Anan City)

Effect of Cytokines on Osteoclast Formation and Bone Resorption during Mechanical Force Loading of the Periodontal Membrane

Mechanical force loading exerts important effects on the skeleton by controlling bone mass and strength. Several in vivo experimental models evaluating the effects of mechanical loading on bone metabolism have been reported. Orthodontic tooth movement is a useful model for understanding the mechanism of bone remodeling induced by mechanical loading. In a mouse model of orthodontic tooth movement, TNF-α was expressed and osteoclasts appeared on the compressed side of the periodontal ligament. In TNF-receptor-deficient mice, there was less tooth movement and osteoclast numbers were lower than in wild-type mice. These results suggest that osteoclast formation and bone resorption caused by loading forces on the periodontal ligament depend on TNF-α. Several cytokines are expressed in the periodontal ligament during orthodontic tooth movement. Studies have found that inflammatory cytokines such as IL-12 and IFN-γ strongly inhibit osteoclast formation and tooth movement. Blocking macrophage colony-stimulating factor by using anti-c-Fms antibody also inhibited osteoclast formation and tooth movement. In this review we describe and discuss the effect of cytokines in the periodontal ligament on osteoclast formation and bone resorption during mechanical force loading

Anti-PD-1 antibody decreases tumour-infiltrating regulatory T cells

Background There are many types of therapies for cancer. In these days, immunotherapies, especially immune checkpoint inhibitors, are focused on. Though many types of immune checkpoint inhibitors are there, the difference of effect and its mechanism are unclear. Some reports suggest the response rate of anti-PD-1 antibody is superior to that of anti-PD-L1 antibody and could potentially produce different mechanisms of action. On the other hand, Treg also express PD-1; however, their relationship remains unclear. Methods In this study, we used osteosarcoma cell lines in vitro and osteosarcoma mouse model in vivo. In vitro, we analyzed the effect of IFN gamma for expression of PD-L1 on the surface of cell lines by flowcytometry. In vivo, murine osteosarcoma cell line LM8 was subcutaneously transplanted into the dorsum of mice. Mouse anti-PD-1 antibody was intraperitoneally administered. we analysed the effect for survival of anti-PD-1 antibody and proportion of T cells in the tumour by flowcytometry. Results We discovered that IFN gamma increased PD-L1 expression on the surface of osteosarcoma cell lines. In assessing the relationship between anti-PD-1 antibody and Treg, we discovered the administration of anti-PD-1 antibody suppresses increases in tumour volume and prolongs overall survival time. In the tumour microenvironment, we found that the administration of anti-PD-1 antibody decreased Treg within the tumour and increased tumour-infiltrating lymphocytes. Conclusions Here we clarify for the first time an additional mechanism of anti-tumour effect-as exerted by anti-PD-1 antibody decreasing Treg- we anticipate that our findings will lead to the development of new methods for cancer treatment.ArticleBMC CANCER. 20(1):25 (2020)journal articl

Organ accumulation and carcinogenicity of highly dispersed multi-walled carbon nanotubes administered intravenously in transgenic rasH2 mice

Purpose: Multiwalled carbon nanotubes (MWCNTs) have been known to enter the circulatory system via the lungs from inhalation exposure; however, its carcinogenicity and subsequent accumulation in other organs have not been adequately reported in the literature. Moreover, the safety of MWCNTs as a biomaterial has remained a matter of debate, particularly when the material enters the circulatory system. To address these problems, we used carcinogenic rasH2 transgenic mice to intravenously administer highly dispersed MWCNTs and to evaluate their carcinogenicity and accumulation in the organs. Methods: Two types of MWCNTs (thin-and thick-MWCNTs) were intravenously administered at a high dose (approximately 0.7 mg per kg body weight) and low dose (approximately 0.07 mg per kg body weight). Results: MWCNTs showed pancreatic accumulation in 3.2% of mice administered with MWCNTs, but there was no accumulation in other organs. In addition, there was no significant difference in the incidence of tumor among the four MWCNTs-administered groups compared to the vehicle group without MWCNTs administration. Blood tests revealed elevated levels in mean red blood cell volume and mean red blood cell hemoglobin level for the MWCNTs-administered group, in addition to an increase in eotaxin. Conclusion: The present study demonstrated that the use of current technology to sufficiently disperse MWCNTs resulted in minimal organ accumulation with no evidence of carcinogenicity.ArticleINTERNATIONAL JOURNAL OF NANOMEDICINE. 14:6465-6480 (2019)journal articl