Recombinant gamma T305A fibrinogen indicates severely impaired fibrin polymerization due to the aberrant function of hole 'a' and calcium binding sites

Introduction: We examined a 6-month-old girl with inherited fibrinogen abnormality and no history of bleeding or thrombosis. Routine coagulation screening tests showed a markedly low level of plasma fibrinogen determined by functional measurement and also a low level by antigenic measurement (functional/antigenic ratio = 0.295), suggesting hypodysfibrinogenemia. Materials and methods: DNA sequence analysis was performed, and gamma T305A fibrinogen was synthesized in Chinese hamster ovary cells based on the results. We then functionally analyzed and compared with that of nearby recombinant gamma N308K fibrinogen. Results: DNA sequence analysis revealed a heterozygous gamma T305A substitution (mature protein residue number). The gamma T305A fibrinogen indicated markedly impaired thrombin-catalyzed fibrin polymerization both in the presence or absence of 1 mM calcium ion compared with that of gamma N308K fibrinogen. Protection of plasmin degradation in the presence of calcium ion or Gly-Pro-Arg-Pro peptide (analogue for so-called knob 'A') and factor XIIIa-catalyzed fibrinogen crosslinking demonstrated that the calcium binding sites, hole 'a' and D:D interaction sites were all markedly impaired, whereas gamma N308K was impaired at the latter two sites. Molecular modeling demonstrated that gamma T305 is localized at a shorter distance than gamma N308 from the high affinity calcium binding site and hole 'a'. Conclusion: Our findings suggest that gamma T305 might be important for construction of the overall structure of the. module of fibrinogen. Substitution of gamma T305A leads to both dysfibrinogenemic and hypofibrinogenemic characterization, namely hypodysfibrinogenemia. We have already reported that recombinant gamma T305A fibrinogen was synthesized normally and secreted slightly, but was significantly reduced.ArticleTHROMBOSIS RESEARCH. 134(2):518-525 (2014)journal articl

Research Trends and Issues Related to Prevention of Infectious Diseases in Early Childhood

本論は，幼児期の健康教育の中でも感染症予防に焦点を当て，近年の研究の動向について，「保育施設における取組みに関する研究」「幼児が実践する感染予防に関する研究」「感染症予防に関する保育者の意識に関する研究」「保育施設と関係機関との連携に関する研究」「保育者養成校における感染症予防教育に関する研究」に分類し考察したものである。幼児を対象とした感染症予防に関する研究成果は，小児保健分野における研究成果が多く，保育実践を対象とした研究は，手洗いに関する論考が散見されるものの，具体的な教育内容や指導法等の実践に直結した研究は未だ進んでいない。感染症対策が強く求められる中で，今後，保育現場の実情に即した様々な観点からの研究の推進が望まれる。In this study, we focused on prevention of infectious diseases (PID) in health education of early childhood, classified into following five researches and discussed recent trends: Research related to 1) approaches in childcare facilities, 2) infection preventive measures undertaken by young children, 3) childcare workers’ consciousness on PID, 4) cooperation between childcare facilities and related institutions, and 5) education for PID in childcare training schools. Many research results in child health have been reported on PID for young children. However, in researches for childcare practice, while handwashing is often discussed, few attempts have been made at a study connected with practices such as concrete educational contents or methods. Because of strong demand for PID, it’s necessary to research from various viewpoints based on the actual circumstances in childcare

Poor Outcome due to the Plasmacytoid Variant of Urothelial Carcinoma

A 72-year-old man visited our hospital due to pollakiuria and lower abdominal pain. Urinary cytology was positive, and cystoscopy revealed diffuse edematous nonpapillary tumor. We performed transurethral biopsy, and clinical stage T3 plasmacytoid variant of urothelial carcinoma (PUC) was diagnosed. Although we planned for radical cystectomy, peritoneal dissemination and lung and pelvic lymph node metastases appeared 3 weeks after the initial visit. We also planned for chemotherapy; however, the metastases rapidly progressed, and he died 7 weeks after the biopsy. PUC is rare and shows an aggressive clinical course and poor prognosis

TRAIL Team Description Paper for RoboCup@Home 2023

Our team, TRAIL, consists of AI/ML laboratory members from The University of Tokyo. We leverage our extensive research experience in state-of-the-art machine learning to build general-purpose in-home service robots. We previously participated in two competitions using Human Support Robot (HSR): RoboCup@Home Japan Open 2020 (DSPL) and World Robot Summit 2020, equivalent to RoboCup World Tournament. Throughout the competitions, we showed that a data-driven approach is effective for performing in-home tasks. Aiming for further development of building a versatile and fast-adaptable system, in RoboCup @Home 2023, we unify three technologies that have recently been evaluated as components in the fields of deep learning and robot learning into a real household robot system. In addition, to stimulate research all over the RoboCup@Home community, we build a platform that manages data collected from each site belonging to the community around the world, taking advantage of the characteristics of the community

The fungal metabolite (+)-terrein abrogates osteoclast differentiation via suppression of the RANKL signaling pathway through NFATc1

Pathophysiological bone resorption is commonly associated with periodontal disease and involves the excessive resorption of bone matrix by activated osteoclasts. Receptor activator of nuclear factor (NF)-κB ligand (RANKL) signaling pathways have been proposed as targets for inhibiting osteoclast differentiation and bone resorption. The fungal secondary metabolite (+)-terrein is a natural compound derived from Aspergillus terreus that has previously shown anti-interleukin-6 properties related to inflammatory bone resorption. However, its effects and molecular mechanism of action on osteoclastogenesis and bone resorption remain unclear. In the present study, we showed that 10 µM synthetic (+)-terrein inhibited RANKL-induced osteoclast formation and bone resorption in a dose-dependent manner and without cytotoxicity. RANKL-induced messenger RNA expression of osteoclast-specific markers including nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), the master regulator of osteoclastogenesis, cathepsin K, tartrate-resistant acid phosphatase (Trap) was completely inhibited by synthetic (+)-terrein treatment. Furthermore, synthetic (+)-terrein decreased RANKL-induced NFATc1 protein expression. This study revealed that synthetic (+)-terrein attenuated osteoclast formation and bone resorption by mediating RANKL signaling pathways, especially NFATc1, and indicated the potential effect of (+)-terrein on inflammatory bone resorption including periodontal disease

The Fungal Metabolite (+)-Terrein Abrogates Ovariectomy-Induced Bone Loss and Receptor Activator of Nuclear Factor-kappa B Ligand-Induced Osteoclastogenesis by Suppressing Protein Kinase-C alpha/beta II Phosphorylation

Osteoporosis is a common disease characterized by a systemic impairment of bone mass and microarchitecture that results in fragility fractures. Severe bone loss due to osteoporosis triggers pathological fractures and consequently decreases the daily life activity and quality of life. Therefore, prevention of osteoporosis has become an important issue to be addressed. We have reported that the fungal secondary metabolite (+)-terrein (TER), a natural compound derived from Aspergillus terreus, has shown receptor activator of nuclear factor-kappa B ligand (RANKL)-induced osteoclast differentiation by suppressing nuclear factor of activated T-cell 1 (NFATc1) expression, a master regulator of osteoclastogenesis. TER has been shown to possess extensive biological and pharmacological benefits; however, its effects on bone metabolism remain unclear. In this study, we investigated the effects of TER on the femoral bone metabolism using a mouse-ovariectomized osteoporosis model (OVX mice) and then on RANKL signal transduction using mouse bone marrow macrophages (mBMMs). In vivo administration of TER significantly improved bone density, bone mass, and trabecular number in OVX mice (p < 0.01). In addition, TER suppressed TRAP and cathepsin-K expression in the tissue sections of OVX mice (p < 0.01). In an in vitro study, TER suppressed RANKL-induced phosphorylation of PKC alpha/beta II, which is involved in the expression of NFATc1 (p < 0.05). The PKC inhibitor, GF109203X, also inhibited RANKL-induced osteoclastogenesis in mBMMs as well as TER. In addition, TER suppressed the expression of osteoclastogenesis-related genes, such as Ocstamp, Dcstamp, Calcr, Atp6v0d2, Oscar, and Itgb3 (p < 0.01). These results provide promising evidence for the potential therapeutic application of TER as a novel treatment compound against osteoporosis

ウィリアム ペインター サク ロミオ ト ジュリエッタ ロン : カイセツ ト ホンヤク

“Romeo and Julietta” is the 25th novel of William Painter\u27s The Palace of Pleasure, Tome 2（1567）, and is one of the sources of William Shakespeare\u27s Romeo and Juliet（1595）. Painter\u27s “Romeo and Julietta” is a minor source compared to Arthur Brooke\u27s The Tragicall Historye of Romeus and Juliet（1562）, which Shakespeare mainly referred to, and therefore has not been paid much attention to so far. This essay first tries to evaluate Painter\u27s work as the second source of Shakespeare\u27s Romeo and Juliet by comparing it with Brooke\u27s and Shakespeare\u27s. Through the comparison of each work\u27s introduction of the story, handling of “Fortune”, characters of Romeo/Romeus and Juliet/Julietta, and the ending of the story, it is obvious how Shakespeare arranged his sources to make his dramatic version effective for the audience\u27s minds. It can also be seen that Shakespeare is not only under the influence of Brooke\u27s long poetic story, but also had the effects of Painter\u27s simple and compact story in mind. The latter part of this essay is a Japanese translation of Painter\u27s “Romeo and Julietta”, which will make it easier to compare Shakespeare\u27s two sources

Physical and functional interaction between DDB and XPA in nucleotide excision repair

Damaged DNA-binding protein (DDB), consisting of DDB1 and DDB2 subunits recognizes a wide spectrum of DNA lesions. DDB is dispensable for in vitro nucleotide excision repair (NER) reaction, but stimulates this reaction especially for cyclobutane pyrimidine dimer (CPD). Here we show that DDB directly interacts with XPA, one of core NER factors, mainly through DDB2 subunit and the amino-acid residues between 185 and 226 in XPA are important for the interaction. Interestingly, the point mutation causing the substitution from Arg-207 to Gly, which was previously identified in a XP-A revertant cell-line XP129, diminished the interaction with DDB in vitro and in vivo. In a defined system containing R207G mutant XPA and other core NER factors, DDB failed to stimulate the excision of CPD, although the mutant XPA was competent for the basal NER reaction. Moreover, in vivo experiments revealed that the mutant XPA is recruited to damaged DNA sites with much less efficiency compared with wild-type XPA and fails to support the enhancement of CPD repair by ectopic expression of DDB2 in SV40-transformed human cells. These results suggest that the physical interaction between DDB and XPA plays an important role in the DDB-mediated NER reaction