Search

Engineering artificial extracellular matrices, based on the biomimicry of the spatial distribution of proteins and growth factors within their native microenvironment, is of great importance for understanding mechanisms of bone tissue regeneration. Herein, photolithography is used to decorate glass surfaces with subcellular patterns of RGD and BMP-2 ligands; two mimetic peptides recognized to be involved in stem cells osteogenesis. The biological relevance of well-defined RGD and BMP-2 patterned surfaces is evaluated by investigating the differentiation of human mesenchymal stem cells (hMSCs) into osteoblasts, in the absence of induction media. The extent of hMSCs differentiation is revealed to be dependent on both the pattern shape and the ligand type. Indeed, the spatial patterning of BMP-2, but not RGD peptide, significantly enhances the extent of hMSCs differentiation, suggesting that geometric cues guide stem cells specification into specialized cells in a ligand type dependent manner. Such cell culture models provide an interesting tool to investigate how stem cells perceive and respond to their microenvironment and may contribute to the development of next-generation biomaterials capable of producing clinically relevant volume of bone tissue. [ABSTRACT FROM AUTHOR]

This study investigated the incidence of sport-related concussion (SRC) in sports, effect of athlete knowledge on reporting behavior differences between collegiate and non-collegiate athletes, and differences in SRC symptoms between sexes and level of participation. In this cross-sectional survey, 1 344 Japanese collegiate and non-collegiate athletes from a single institute were analyzed. Using a web-based survey, demographics, general SRC, knowledge of SRC, the most recent SRC reporting behaviors, and symptom presentation were examined. The prevalence of SRC during the academic year 2016-2017 was 2.68 (95% confidence interval [ CI ]: 1.88-3.69) across all sports. The prevalence of SRC was 33.3 (95% CI : 17.96-51.83) in rugby union and 8.33 (95% CI : 1.03-27.00) in women's soccer. The prevalence of SRC in males (3.47 [95% CI : 2.38-4.86] was 3.65 times higher than that in females (0.95 [95% CI : 0.26-2.41]). In total, the mean total score of knowledge was 5.30 (4.2) across 25 questions; dizziness was the most well-known symptom (867/1 344, 64.5%), followed by headache (59.3%). Being more emotional (44/1 345, 3.3%) was the least frequently known symptom. Level of participation did not affect scores (5.16 [3.96] vs. 5.52 [4.54]; p  ​= ​0.131). All 87 disclosing participants experienced drowsiness and irritability and felt more emotional. In terms of sex and participant level, no significant differences were found in any symptoms. This study found very low rates of concussion education in Japan. Dissemination of concussion education is essential in the future to recognize concussion earlier and prevent severe concussive injury., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Chengdu Sport University. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

Objectives: Graves' disease (GD) has been highlighted as a possible adverse effect of the respiratory syndrome coronavirus-2 (SARS-CoV-2) vaccine. However, it is unknown if the SARS-CoV-2 vaccine disrupts thyroid autoimmunity. We aimed to present long-term follow-up of thyroid autoimmunity after the SARS-CoV-2 BNT162b2 mRNA vaccine., Methods: Serum samples collected from seventy Japanese healthcare workers at baseline, 32 weeks after the second dose (pre-third dose), and 4 weeks after the third dose of the vaccine were analyzed. The time courses of anti-SARS-CoV-2 spike immunoglobulin G (IgG) antibody, thyroid-stimulating hormone receptor antibody (TRAb), and thyroid function were evaluated. Anti-thyroglobulin antibodies (TgAb) and anti-thyroid peroxidase antibodies (TPOAb) were additionally evaluated in thirty-three participants., Results: The median age was 50 (IQR, 38-54) years and 69% were female. The median anti-spike IgG antibody titer was 17627 (IQR, 10898-24175) U/mL 4 weeks after the third dose. The mean TRAb was significantly increased from 0.81 (SD, 0.05) IU/L at baseline to 0.97 (SD, 0.30) IU/L 4 weeks after the third dose without functional changes. An increase in TRAb was positively associated with female sex (β = 0.32, P = 0.008) and low basal FT4 (β = -0.29, P = 0.02) and FT3 (β = -0.33, P = 0.004). TgAb was increased by the third dose. Increase in TgAb was associated with history of the thyroid diseases (β = 0.55, P < 0.001)., Conclusions: SARS-CoV-2 BNT162b2 mRNA vaccine can disrupt thyroid autoimmunity. Clinicians should consider the possibility that the SARS-CoV-2 vaccine may disrupt thyroid autoimmunity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Morita, Takagi, Inaba, Furukawa, Kishimoto, Uraki, Shimo, Takeshima, Uraki, Doi, Imagawa, Kokawa, Konami, Hara, Hara, Sone, Furuta, Nishi, Doi, Tamura and Mastuoka.)

Objective: To describe neurological outcomes after sudden cardiac arrests (SCAs) in road and long-distance races using a rapid mobile automated external defibrillator system (RMAEDS) intervention., Methods: A total of 42 SCAs from 3 214 701 runners in 334 road and long-distance races from 1 February 2007 to 29 February 2020 were examined. Demographics, SCA interventions, EMS-related data and SCA-related outcomes were measured. Primary endpoints were favourable neurological outcomes (Cerebral Performance Categories 1-2) at 1-month and 1-year post-SCA. Secondary endpoints were factors related to the field return of spontaneous circulation (ROSC) and resuscitation characteristics, including the initial ECG waveform classification and resuscitation sequence times according to the initial ECG rhythm., Results: The SCA incidence rate was 1.31 per 100 000 runners (age: median (IQR), 51 (36.5, 58.3) years). Field ROSC and full neurological recovery at 1-month post-SCA was achieved 90.4% and 92.9% of cases, respectively. In 22 cases in which bystander cardiopulmonary resuscitation was initiated within 1 min and defibrillation performed within 3 min, full neurological recovery was achieved at 1-month and 1-year post-SCA in 95.5.% and 95.5% of cases, respectively., Conclusions: The RMAEDS successfully treated patients with SCA during road and long-distance races yielding a high survival rate and favourable neurological outcomes. These findings support rapid intervention and the proper placement of healthcare teams along the race course to initiate chest compressions within 1 min and perform defibrillation within 3 min., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

We recently found that the membrane-bound receptor activator of NF-κB ligand (RANKL) on osteoblasts works as a receptor to stimulate osteoblast differentiation, however, the reason why the RANKL-binding molecules stimulate osteoblast differentiation has not been well clarified. Since the induction of cell-surface receptor clustering is known to lead to cell activation, we hypothesized that the induction of membrane-RANKL clustering on osteoblasts might stimulate osteoblast differentiation. Immunoblotting showed that the amount of RANKL on the membrane was increased by the RANKL-binding peptide OP3-4, but not by osteoprotegerin (OPG), the other RANKL-binding molecule, in Gfp-Rankl-transfected ST2 cells. Observation under a high-speed atomic force microscope (HS-AFM) revealed that RANKL molecules have the ability to form clusters. The induction of membrane-RANKL-OPG-Fc complex clustering by the addition of IgM in Gfp-Rankl-transfected ST2 cells could enhance the expression of early markers of osteoblast differentiation to the same extent as OP3-4, while OPG-Fc alone could not. These results suggest that the clustering-formation of membrane-RANKL on osteoblasts could stimulate early osteoblast differentiation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

We recently found that the receptor activator of NF-κB ligand (RANKL)-binding peptide, OP3-4 stimulated the differentiation of both chondrocytes and osteoblasts. OP3-4 is also shown to inhibit cartilage degeneration. To clarify whether the peptide can inhibit cartilage degeneration without stimulating bone formation, we first performed a proliferation assay using C3H10T1/2 (the murine mesenchymal stem cell line), which is the common origin of both chondrocytes and osteoblasts. The RANKL-binding peptides, OP3-4 and W9, promoted cellular proliferation at 24 and 48 h, respectively. Next, we injected both peptides into the intra-articular space of the knee joints of mice with monosodium-iodoacetate (MIA)-induced osteoarthritis to clarify the effects of the peptides on cartilage tissue. Twenty-five nine-week-old male C57BL/6J mice received injections of vehicle, or the same molar amount of W9, OP3-4, or a control peptide (which could not stimulate osteoblast differentiation) on days 7, 14, and 21 after the injection of MIA. The mice were sacrificed on day 28. The histomorphometric analyses revealed that both peptides inhibited the degeneration of cartilage without enhancing bone formation activity. Our data suggest that the stimulation of mesenchymal cell proliferation by the RANKL-binding peptides might lead to the inhibition of cartilage degeneration., (Copyright © 2017 The Authors. Production and hosting by Elsevier B.V. All rights reserved.)

The thermal stability of oxidized single-walled carbon nanotubes (SWNTs) with various degrees of oxidation was investigated. The oxidized SWNTs exhibited lower absorption and radial breathing mode (RBM) peaks and a higher intensity ratio of the D band to the G band (D/G) in their absorption and Raman spectra than those of the pristine SWNTs. After the thermal treatment, the D/G ratio of the oxidized SWNTs almost recovered its original intensity, regardless of the degree of oxidation. The absorption, photoluminescence (PL), and RBM peaks could not recover their original intensities when the oxidation degree was high. The results indicate that the elimination and decomposition reactions proceeded competitively depending on the degree of oxidation. In addition, a new PL peak was observed in the near-infrared region, and the PL peak intensity increased with the subsequent thermal treatment. The theoretical calculations provided an insight into the possible pathways for the decomposition of oxidized SWNTs, showing that the O 2 elimination and CO/CO 2 evolution proceed competitively during thermal treatment., (© 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Unlabelled: Human bone marrow mesenchymal stem cells (hBMSCs) commitment and differentiation are dictated by bioactive molecules sequestered within their Extra Cellular Matrix (ECM). One common approach to mimic the physiological environment is to functionalize biomaterial surfaces with ECM-derived peptides able to recruit stem cells and trigger their linage-specific differentiation. The objective of this work was to investigate the effect of RGD and BMP-2 ligands crosstalk and density on the extent of hBMSCs osteogenic commitment, without recourse to differentiation medium. RGD peptide promotes cell adhesion via cell transmembrane integrin receptors, while BMP-2 peptide, corresponding to residues 73-92 of Bone Morphogenetic Protein-2, was shown to induce hBMSCs osteoblast differentiation. The immobilization of peptides on aminated glass was ascertained by X-ray Photoelectron Spectroscopy (XPS), the density of grafted peptides was quantified by fluorescence microscopy and the surface roughness was evaluated using Atomic Force Microscopy (AFM). The osteogenic commitment of hBMSCs cultured on RGD and/or BMP-2 surfaces was characterized by immunohistochemistry using STRO-1 as specific stem cells marker and Runx-2 as an earlier osteogenic marker. Biological results showed that the osteogenic commitment of hBMSCs was enhanced on bifunctionalized surfaces as compared to surfaces containing BMP-2, while on RGD surfaces cells mainly preserved their stemness character. These results demonstrated that RGD and BMP-2 mimetic peptides act synergistically to enhance hBMSCs osteogenesis without supplementing the media with osteogenic factors. These findings contribute to the development of biomimetic materials, allowing a deeper understanding of signaling pathways that govern the transition of stem cells towards the osteoblastic lineage., Statement of Significance: For a long time, scientists thought that the differentiation of Mesenchymal Stem Cells (MSCs) into bone cells was dictated by growth factors. This manuscript shed light on other ligands that play a crucial role in regulating MSCs fate. In concrete terms, it was demonstrated that the osteoinductive effect of BMP-2 peptide is 2 folds improved in the presence of adhesive RGD peptide. Compared to previous works highlighting this synergistic cooperation between RGD and BMP-2 peptides, the main strength of this work lies to the use of primitive human cells (hMSCs) and well-defined biomimetic material surfaces (controlled surface roughness and peptide densities). This work provides valuable insights to develop custom-designed in vitro cell culture models, capable of targeting the desired cell response., (Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Treatment of CuCl2 with a bis-benzimidazole-type ligand, p-bbiteb, afforded a new 2D coordination polymer bearing cationic Cu2L4 cages on the sheet framework. The regular arrangement of the cages created channel-like space for anions. The compound removed ClO4(-) and NO3(-) from aqueous solutions by exchange with Cl(-) in the channels.

Bone sialoprotein (BSP) is a multifunctional extracellular matrix protein found in mineralized tissues, including bone, cartilage, tooth root cementum (both acellular and cellular types), and dentin. In order to define the role BSP plays in the process of biomineralization of these tissues, we analyzed cementogenesis, dentinogenesis, and osteogenesis (intramembranous and endochondral) in craniofacial bone in Bsp null mice and wild-type (WT) controls over a developmental period (1-60 days post natal; dpn) by histology, immunohistochemistry, undecalcified histochemistry, microcomputed tomography (microCT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and quantitative PCR (qPCR). Regions of intramembranous ossification in the alveolus, mandible, and calvaria presented delayed mineralization and osteoid accumulation, assessed by von Kossa and Goldner's trichrome stains at 1 and 14 dpn. Moreover, Bsp(-/-) mice featured increased cranial suture size at the early time point, 1 dpn. Immunostaining and PCR demonstrated that osteoblast markers, osterix, alkaline phosphatase, and osteopontin were unchanged in Bsp null mandibles compared to WT. Bsp(-/-) mouse molars featured a lack of functional acellular cementum formation by histology, SEM, and TEM, and subsequent loss of Sharpey's collagen fiber insertion into the tooth root structure. Bsp(-/-) mouse alveolar and mandibular bone featured equivalent or fewer osteoclasts at early ages (1 and 14 dpn), however, increased RANKL immunostaining and mRNA, and significantly increased number of osteoclast-like cells (2-5 fold) were found at later ages (26 and 60 dpn), corresponding to periodontal breakdown and severe alveolar bone resorption observed following molar teeth entering occlusion. Dentin formation was unperturbed in Bsp(-/-) mouse molars, with no delay in mineralization, no alteration in dentin dimensions, and no differences in odontoblast markers analyzed. No defects were identified in endochondral ossification in the cranial base, and craniofacial morphology was unaffected in Bsp(-/-) mice. These analyses confirm a critical role for BSP in processes of cementogenesis and intramembranous ossification of craniofacial bone, whereas endochondral ossification in the cranial base was minimally affected and dentinogenesis was normal in Bsp(-/-) molar teeth. Dissimilar effects of loss of BSP on mineralization of dental and craniofacial tissues suggest local differences in the role of BSP and/or yet to be defined interactions with site-specific factors., (Published by Elsevier Inc.)

Little is known about the role of cell-cell adhesion in the development of mineralized tissues. Here we report that PERP, a tetraspan membrane protein essential for epithelial integrity, regulates enamel formation. PERP is necessary for proper cell attachment and gene expression during tooth development, and its expression is controlled by P63, a master regulator of stratified epithelial development. During enamel formation, PERP is localized to the interface between the enamel-producing ameloblasts and the stratum intermedium (SI), a layer of cells subjacent to the ameloblasts. Perp-null mice display dramatic enamel defects, which are caused, in part, by the detachment of ameloblasts from the SI. Microarray analysis comparing gene expression in teeth of wild-type and Perp-null mice identified several differentially expressed genes during enamel formation. Analysis of these genes in ameloblast-derived LS8 cells upon knockdown of PERP confirmed the role for PERP in the regulation of gene expression. Together, our data show that PERP is necessary for the integrity of the ameloblast-SI interface and that a lack of Perp causes downregulation of genes that are required for proper enamel formation.

Herpesviral infections have been documented in some cetaceans; however, they have not yet been identified in species in the western North Pacific. In the present study, 178 tissue samples from 76 stranded cetacean individuals were tested for the presence of herpesviruses. Herpesvirus genomic DNA fragments surrounding the DNA polymerase gene were amplified in samples from four individuals. TA cloning and direct sequencing of these DNA fragments revealed the presence of two novel alphaherpesviruses, and two novel gammaherpesviruses in the four cetacean individuals. The alphaherpesviruses were associated with the lung tissue of a false killer whale (Pseudorca crassidens), and with the mucus of a melon-headed whale (Peponocephala electra). The gammaherpesviruses were found in the lymph tissues of a Stejneger's beaked whale (Mesoplodon stejnegeri) and a sperm whale (Physeter macrocephalus). The phylogenetic tree using amino acid sequences of the DNA polymerase gene supported the inclusion of the novel viruses identified here in a single monophyletic group containing alphaherpesviruses from other Atlantic cetacean species. Conversely, the novel gammaherpesviruses formed an independent clade distant from other known cetacean gammaherpesviruses.

Aim: We reevaluated waist circumference as a diagnostic criterion of metabolic syndrome (MetS) in Japanese., Methods: We enrolled 5,571 subjects (3,148 men and 2,423 women) who had health check-ups in our center. The criterion was reevaluated using the positive predictive value of a receiver-operating characteristics (ROC) curve at 10 different hypothesized lengths of waist circumference with or without a cluster of risk factors. We also drew ROC curves based on the atherosclerotic findings of clinical examinations., Results: Based on the ROC curves, the optimal waist circumference cut-off was 85 cm in men and 80 cm in women. Using this 80 cm cut-off point in women, misdiagnosis rates of MetS were lowered (-19.1--56.6%) compared to the cut-off point currently in use. Integrating the influence of height, namely by using a waist-to-height(2) ratio, misdiagnosis rates in shorter populations were decreased in both men and women., Conclusion: These data suggested an optimal waist circumference cut-off to improve the diagnostic probability of MetS in Japanese women of 80 cm, as well as the utility of an easily detected anthropometric index such as a waist-to-height (cm x 100/cm) or waist-to-height(2) (cm x 10,000/cm(2)) ratio, determined as 51 in men and 52 in women, or 30 in men and 33 in women, respectively.

The amino-acid sequences of the T-domain region of the Tbx4 gene, which is required for hindlimb development, are 100% identical in humans and mice. Cetaceans have lost most of their hindlimb structure, although hindlimb buds are present in very early cetacean embryos. To examine whether the Tbx4 gene has the same function in cetaceans as in other mammals, we analyzed Tbx4 sequences from cetaceans, dugong, artiodactyls and marine carnivores. A total of 39 primers were designed using human and dog Tbx4 nucleotide sequences. Exons 3, 4, 5, 6, 7, and 8 of the Tbx4 genes from cetaceans, artiodactyls, and marine carnivores were sequenced. Non-synonymous substitution sites were detected in the T-domain regions from some cetacean species, but were not detected in those from artiodactyls, the dugong, or the carnivores. The C-terminal regions contained a number of non-synonymous substitutions. Although some indels were present, they were in groups of three nucleotides and therefore did not cause frame shifts. The dN/dS values for the T-domain and C-terminal regions of the cetacean and artiodactylous Tbx4 genes were much lower than 1, indicating that the Tbx4 gene maintains it function in cetaceans, although full expression leading to hindlimb development is suppressed.

In a screening study on immunomodulatory fungal constituents, eleven new 2-pyrones tentatively named TT-1 (1), and TT-2-11 (2-11) have been isolated from a Fungi Imperfecti, Trichurus terrophilus, and designated rasfonin (1), and trichurusin B-K (2-11), respectively. Compounds 1-4 exhibited considerably high immunosuppressive activities, and compounds 8-11 have shown moderate ones. The structure-activity relationships of these constituents have also been discussed.

Information about letters and the physical structure of language printed in Roman characters was given to children beginning to read. Experimental investigations coupled three alternative graphic modes of printing upper- and lower-case letters with an instructional intervention termed "Alpha-Beta" which provides practice in letter sorting, matching of letters, associative matching, and memory matching. In respect to graphics, Mode A letters were in standard alphabet form. Mode B provided standard letters with each backed by a unique half-tone (Visually Stippled Alphabet); Mode C provided standard letters with each backed by a unique visual texture (Visually Patterned Alphabet). Pre-posttest change in reading readiness was measured using the Metropolitan Readiness Test. In the first study 224 English-speaking 5- to 6-yr.-old children were tested. In the second there were 158 Spanish-speaking girls and boys 6 to 7 years old. It was predicted that Alpha-Beta intervention involving visually patterned alphabet would lead to the greatest increases in readiness scores. This is confirmed in both studies for children low in reading readiness preexperiment. Children high in reading readiness are less affected. The second experiment involved Spanish-speaking children and investigated intervention by Alpha-Beta against a no-intervention control. This confirms the value of Alpha-Beta per se. Possible explanations for the improvements are identified.