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The roles of CD4+T helper cells (TH) in shaping localized memory B and CD8+T cell immunity in the mucosal tissues are largely unexplored. Here, we report that lung THcells provide local assistance for the optimal development of tissue-resident memory B (BRM) and CD8+T (TRM) cells following the resolution of primary influenza virus infection. We identify a population of tissue-resident CD4+TH(akaTRH) cells that co-exhibit follicular T helper (TFH) and TRMcell features and mediate local help of CD4+T cells to B and CD8+T cells. Optimal TRHcell formation requires lung B cells and transcription factors involved in TFHor TRMdevelopment. Further, we show that TRHcells deliver local help to B and CD8 T cells through CD40L and IL-21-dependent mechanisms. Our data have uncovered a new tissue-resident THcell population that is specialized in assisting the development of mucosal protective B and CD8+T cell responsesin situ.

In 2017, the second national reference standard (NRS) for Gloydius snake venom was established to replace the first NRS for Gloydius snake venom. In connection with the second venom NRS, a candidate for the first NRS for Gloydius snake antivenom was produced in 2017. In this study, the qualification of the candidate was estimated and the potency was determined by a collaborative study. The potency (anti-lethal titer and anti-hemorrhagic titer) of the candidate was determined by measuring the capability of the antivenom to neutralize the lethal and hemorrhagic effects of the second NRS for Gloydius snake venom, which was calibrated against the regional reference standard for Gloydius snake antivenom established in 2006. Two Korean facilities contributed data from 20 independent assays. Subsequently, one foreign national control research laboratories participated in this collaborative study. The general common potency of the anti-lethal and anti-hemorrhagic titers was obtained from the results of a total of 25 tests performed at three facilities. According to the results of the present study, the candidate preparation showed good quality and is judged to be suitable to serve as the first NRS for Gloydius snake antivenom with the following potency: an anti-lethal titer of 3100 unit (U) (95% confidence interval 2991–3276 U) and anti-hemorrhagic titer of 3000 U (95% confidence interval 2849–3159 U). In conclusion, the first NRS for Gloydius snake antivenom was established in this study. This reference standard will be used routinely for quality control of a snake antivenom product by manufacturer in Korea, which also can be used for national quality control, including a national lot-release test of the snake antivenom product.

Memory B cells are a key cellular component of the protective humoral responses to infectious pathogens. Most of our knowledge of memory B-cell responses comes from studies using mono-epitopic model antigens that elicit systemic humoral responses dominated by canonical B-cell antigen receptors. This approach successfully dissected the systemic responses of memory B cells and greatly advanced our understanding of memory B-cell formation, maintenance, and reactivation to re-invading antigens in the secondary lymphoid organs. However, the canonical memory B-cell responses fail to fully recapitulate the heterogeneity of the protective memory responses. Indeed, accumulating studies using “natural” antigens and live pathogens have uncovered new aspects of memory B-cell responses, which are achieved by memory B cells with different phenotypes, tissue residence, and responsiveness to antigen stimulation. Such non-canonical memory B-cell responses are frequently observed in local sites where live pathogens initially infect and replicate. Importantly, the local memory B-cell responses often serve as the first line of defense against re-infecting pathogens, thereby playing an essential role in controlling the pathogens. Here, we provide a comprehensive overview of the systemic and local memory B-cell responses in the humoral protective immunity against pathogens.

As a sustainable ecosystem, the general firing process for ceramics emits large amounts of CO2 gas; thus in ceramics production, the focus is the nonfiring process; however, the solidification and strengthen mechanism of this nonfiring system, which essentially reacts between surface-activated ceramic particles and a solvent, has not been elucidated to date. The nonfiring process had three steps, i.e., particle surface activate process by grinding process, maintaining the active state until starting nonfiring solidification begins, and nonfiring solidification process. Thus, in this study, the reaction of silica and water was simulated by adapting molecular dynamics based on LAMMPS with ReaxFF potentials. Reproducing the activated silica surface state, three ended models called O model, Si model, and OH model were prepared which indicated ended molecules of each surface. These models and a water molecule as a solvent were bonded in the atomic scale, and the energetic state and mechanical properties were evaluated. A reacted or structured O-H-O bond was reproduced in the nonfiring process in the O-ended model. The bond was a hydrogen bond. A Si-O-Si bond was produced when a Si atom was ended on the interface. The bonded interface was able to tensile. However, the tensile strength was weaker than that of the solid silica model. The nonbonded OH model did not have tensile strength.

Sialic acids are unique sugars with negative charge and exert various biological functions such as regulation of immune systems, maintenance of nerve tissues and expression of malignant properties of cancers. Alpha 2,6 sialylated N-glycans, one of representative sialylation forms, are synthesized by St6gal1 or St6gal2 gene products in humans and mice. Previously, it has been reported that St6gal1 gene is ubiquitously expressed in almost all tissues. On the other hand, St6gal2 gene is expressed mainly in the embryonic and perinatal stages of brain tissues. However, roles of St6gal2 gene have not been clarified. Expression profiles of N-glycans with terminal α2,6 sialic acid generated by St6gal gene products in the brain have never been directly studied. Using conventional lectin blotting and novel sialic acid linkage-specific alkylamidationmass spectrometry method (SALSA-MS), we investigated the function and expression of St6gal genes and profiles of their products in the adult mouse brain by establishing KO mice lacking St6gal1 gene, St6gal2 gene, or both of them (double knockout). Consequently, α2,6-sialylated N-glycans were scarcely detected in adult mouse brain tissues, and a majority of α2,6-sialylated glycans found in the mouse brain were O-linked glycans. The majority of these α2,6-sialylated O-glycans were shown to be disialyl-T antigen and sialyl-(6)T antigen by mass spectrometry analysis. Moreover, it was revealed that a few α2,6-sialylated N-glycans were produced by the action of St6gal1 gene, despite both St6gal1 and St6gal2 genes being expressed in the adult mouse brain. In the future, where and how sialylated O-linked glycoproteins function in the brain tissue remains to be clarified.

As for magnesium (Mg) alloys, it has been noted that they are inferior to plastic deformation, but improvement in the mechanical properties by further refinement of grain size has been recently suggested. It means the importance of atomistic view of polycrystalline interface of Mg crystal. In this study, to discuss the deformation mechanism of polycrystalline Mg, atomistic grain boundary (GB) models by using coincidence site lattice (CSL) theory are constructed and are simulated for their relaxed and deformatted structures. First, GB structures in which the axis of rotation is in [11¯00] direction are relaxed at 10 Kelvin, and the GB energies are evaluated. Then, the deformation mechanism of each GB model under uniaxial tensile loading is observed by using the molecular dynamics (MD) method. The present MD simulations are based on embedded atom method (EAM) potential for Mg crystal. As a result, we were able to observe atomistically a variety of GB structures and to recognize significant difference in deformation mechanism between low-angle GBs and high-angle GBs. A close scrutiny is made on phenomena of dislocation emission processes peculiar to each atomistic local structure in high-angle GBs.

Virus-like particles (VLPs) provide a well-established vaccine platform; however, the immunogenic properties acquired by VLP structure remain poorly understood. In this study, we showed that systemic vaccination with norovirus VLP recalls human IgA responses at higher magnitudes than IgG responses under a humanized mouse model that was established by introducing human PBMCs in severely immunodeficient mice. The recall responses elicited by VLP vaccines depended on VLP structure and the disruption of VLP attenuated recall responses, with a more profound reduction being observed in IgA responses. The IgA-focusing property was also conserved in a murine norovirus-primed model under which murine IgA responses were recalled in a manner dependent on VLP structure. Importantly, the VLP-driven IgA response preferentially targeted virus-neutralizing epitopes located in the receptor-binding domain. Consequently, VLP-driven IgA responses were qualitatively superior to IgG responses in terms of the virus-neutralizing activity in vitro. Furthermore, the IgA in mucosa obtained remarkable protective function toward orally administrated virus in vivo. Thus, our results indicate the immune-focusing properties of the VLP vaccine that improve the quality/quantity of mucosal IgA responses, a finding with important implications for developing mucosal vaccines.

Abaloparatide (ABL) is a novel synthetic peptide analog of parathyroid hormone-related protein. In previous reports, intermittent ABL administration showed robust bone mineral density (BMD) increase and reduced the incidence of fractures in patients with osteoporosis, while its calcemic effect was reduced, as compared with teriparatide (TPTD), a parathyroid hormone N-terminal fragment. The present study aimed to elucidate the effects of ABL on bone anabolism and bone turnover as compared with TPTD. In ovariectomized (OVX) rats, ABL increased the bone strength and BMD of lumbar spine by intermittent administration similar to TPTD. Both ABL and TPTD increased the bone formation marker serum P1NP with little effect on the bone resorption maker urine DPD/Cr, suggesting anabolic effects on bone. In human osteoblastic cells, both peptides increased the expression of bone resorption-related factors such as RANKL/OPG and M-CSF, and the effects of ABL were significantly attenuated as compared with those of TPTD under transient 6-h treatment, although no significant differences were found under continuous treatment. In contrast, ABL and TPTD similarly promoted the expression of bone formation-related factors, IGF-1 and osteocalcin. In addition, there were no significant differences in the effects on WNT signaling inhibitors such as sclerostin and dickkopf-related protein 1 (DKK1) between the two peptides. These results demonstrate that ABL exerts bone anabolic effects in OVX rats. It is also indicated that ABL stimulates the expression of RANKL/OPG and M-CSF less than TPTD, while showing similar effects on bone formation-related factors and WNT signaling inhibitors in vitro. The profile of ABL indicates that it would be a suitable bone anabolic agent for osteoporosis.

The strength against fracture initiation from an interface free-edge of bonded copper (Cu) and silicon nitride (SiN) micro-components is studied. Quantitative fracture tests are conducted either in a vacuum environment or in a gaseous hydrogen (H $$_{2})$$ environment. By using two kinds of specimens, Types I and II, having the same free-edge geometry (135 $$^{\circ }$$ /135 $$^{\circ }$$ ) but different outer shape, the validity of the linear fracture mechanics criterion is critically evaluated. For specimens tested in vacuum, fracture initiation strength represented by the elastic stress intensity factor of the near-edge normal stress component is found to be the same irrespective of the specimen type, indicating that the fracture mechanics criterion holds. For specimens tested in H $$_{2}$$ gas, the strength of both types is reduced and, interestingly, Type II strength shows greater reduction than Type I. In other words, the fracture mechanics criterion does not hold in H $$_{2}$$ gas. Such a difference may be attributable to the difference of local plasticity in each specimen type.

The fatigue properties of 6061-T6 aluminum (Al) alloy containing additional solute magnesium (Mg) is studied. As previously reported, this special alloy exhibits eminent strain-aging characteristics which is absent in a normal 6061-T6 alloy with a stoichiometric Mg2Si composition. In this study, particular attention is paid to the environmental effect on the fatigue properties of this new alloy. The S-N curves of the alloys are compared in different environments (ambient air, dry air, dry nitrogen). A clear knee-point (fatigue limit), which is atypical to the normal 6061-T6 alloy, appears for the new alloy irrespective of the environment. The fatigue limit is shown to be controlled by the threshold against small crack propagation in all environments. On the other hand, the coaxing effect, the manifestation of a time-dependent strengthening mechanism at the non-propagating crack tip, is clearly observed in the ambient air.

The time-dependent crack initiation from the interface free-edge of adhesively bonded axisymmetric columnar butt joints (epoxy/SUS, edge shape: 90°/90°) was investigated in detail. With the change of applied stress level, σn, the butt joints exhibited crack initiation life, tC, that varied about four orders of seconds (102–106 s). Such a clear time-dependent life property was then studied in terms of the fracture mechanics. The near-edge stress/strain field at the crack initiation was numerically evaluated with the finite element method (FEM) by applying the time-hardening creep law to the epoxy resin. It was found that the critical asymptotic stress field along the interface represented by the combination of two parameters, λ σ cr (creep stress singularity index) and K σ cr (creep stress intensity factor), satisfies a unique relation irrespective of (σn, tC) sets. The same tendency was also confirmed when the near-edge total strain field parameters were employed. These results indicate that the K-λ criterion, originally developed for static fracture problems, still holds its validity in the time domain.

Potently neutralizing SARS-CoV-2 antibodies often target the spike protein receptor binding site (RBS), but the variability of RBS epitopes hampers broad neutralization of multiple sarbecoviruses and drifted viruses. Here, using humanized mice, we identified an RBS antibody with a germline VH gene that potently neutralized SARS-related coronaviruses including SARS-CoV and SARS-CoV-2 variants. X-ray crystallography revealed coordinated recognition by the heavy chain of non-RBS conserved sites and the light chain of RBS with a binding angle mimicking the ACE2 receptor. The minimum footprints in the hypervariable region of RBS contributed to the breadth of neutralization, which was enhanced by IgG3 class switching. The coordinated binding resulted in broad neutralization of SARS-CoV and emerging SARS-CoV-2 variants of concern. Low dose therapeutic antibody treatment in hamsters reduced the virus titers and morbidity during SARS-CoV-2 challenge. The structural basis for broadly neutralizing activity may inform the design of broad spectrum of therapeutics and vaccines., Graphical Abstract, Antigenic variability in SARS-related coronavirus decreases the likelihood of cross-neutralization by monoclonal antibodies. Onodera et al. identify a broadly neutralizing SARS-CoV-2 antibody in humanized mice that targets a vulnerable site of SARS-CoV and SARS-CoV-2 variants. Structural analysis reveals that the broad neutralization is coordinated by the heavy and light chains and is enhanced by IgG3 class switching.

An expanded myeloid cell compartment is a hallmark of severe coronavirus disease 2019 (COVID-19). However, data regarding myeloid cell expansion have been collected in Europe, where the mortality rate by COVID-19 is greater than those in other regions including Japan. Thus, characteristics of COVID-19-induced myeloid cell subsets remain largely unknown in the regions with low mortality rates. Here, we analyzed cellular dynamics of myeloid-derived suppressor cell (MDSC) subsets and examined whether any of them correlate with disease severity and prognosis, using blood samples from Japanese COVID-19 patients. We observed that polymorphonuclear (PMN)-MDSCs, but not other MDSC subsets, transiently expanded in severe cases but not in mild or moderate cases. Contrary to previous studies in Europe, this subset selectively expanded in survivors of severe cases and subsided before discharge, but such transient expansion was not observed in non-survivors in Japanese cohort. Analysis of plasma cytokine/chemokine levels revealed positive correlation of PMN-MDSC frequencies with interleukin 8 (IL-8) levels prior to the cell expansion, indicating the involvement of IL-8 on recruitment of PMN-MDSCs to peripheral blood following the onset of severe COVID-19. Thus, our data indicates that transient expansion of the PMN-MDSC subset results in improved clinical outcome. Thus, this myeloid cell subset may be a predictor of prognosis in cases of severe COVID-19 in Japan.

Virus-specific memory B cells (Bmem) play a crucial role in protecting against variant viruses. The ability to recognize these variant viruses, defined as antibody breadth, is achieved in Bmem populations by two very different pathways, germline-encoded cross-reactivity and affinity-driven, somatic evolution in germinal centers (GCs) for conserved viral epitopes. The latter class of broadly-reactive Bmem cells are not cross-reactive per se, but bind epitopes crucial for viral fitness. Although these conserved epitopes are often weakly immunogenic, the GC reaction is surprisingly permissive for the continued survival/proliferation of B cells that bind with low affinity or react to cryptic epitopes, increasing their chance of memory recruitment. In this review, we discuss the adaptive strategies of B-cell memory to viral antigenic variations.

Influenza vaccines of H7N9 subtype are consistently less immunogenic in humans than vaccines developed for other subtypes. Although prior immunoinformatic analysis identified T-cell epitopes in H7 hemagglutinin (HA) which potentially enhance regulatory T cell response due to conservation with the human genome, the links between the T-cell epitopes and low immunogenicity of H7 HA remains unknown due to the lack of animal models reproducing the response observed in humans. Here, we utilized a humanized mouse model to recapitulate the low immunogenicity of H7 HA. Our analysis demonstrated that modification of a single H7 epitope by changing 3 amino acids so that it is homologous with a known H3 immunogenic epitope sequence significantly improved the immunogenicity of the H7 HA in the humanized mouse model, leading to a greater than 4-fold increase in HA-binding IgG responses. Thus, we provide experimental evidence for the important contribution of this H7-specific T cell epitope in determining the immunogenicity of an influenza vaccine. Furthermore, this study delineates strategies that can be used for screening and selecting vaccine strains using immunoinformatics tools and a humanized mouse model.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Objective Resting CD4+ T cells are major reservoirs of latent HIV-1 infection, and may be formed during the early phase of the infection. Although CCR5-tropic (R5) HIV-1 is highly transmissible during the early phase, newly infected individuals have usually been exposed to a mixture of R5 and CXCR4-tropic (X4) viruses, and X4 viral DNA is also detectable in the host. Our aim was to identify which subsets of resting CD4+ T cells contribute to forming the latent reservoir in the presence of both X4 and R5 viruses. Results Primary resting CD4+ naïve T (TN) cells, CCR5− memory T (TM) cells, and CCR5+ TM cells isolated by flow cytometry were infected simultaneously with X4 and R5 HIV-1, which harbored different reporter genes, and were cultured in the resting condition. Flow cytometry at 3 days post-infection demonstrated that X4 HIV-1+ cells were present in all three subsets of cells, whereas R5 HIV-1+ cells were present preferentially in CCR5+ TM cells, but not in TN cells. Following CD3/CD28-mediated activation at 3 days post-infection, numbers of R5 HIV-1+ cells and X4 HIV-1+ cells increased significantly only in the CCR5+ TM subset, suggesting that it provides a major reservoir of replication-competent, latently infected viruses. Electronic supplementary material The online version of this article (10.1186/s13104-019-4281-5) contains supplementary material, which is available to authorized users.

Immunoglobulins play important roles in antigen recognition during the immune response, and the complementarity-determining region (CDR) 3 of the heavy chain is considered as the critical antigen-binding site. We previously developed a statistical protocol for the extensive analysis of heavy chain variable region repertoires and the dynamics of their immune response using next-generation sequencing (NGS). The properties of important antibody heavy chains predicted in silico by the protocol were examined by gene synthesis and antibody protein expression; however, the corresponding light chain that matches with the heavy chain could not be predicted by our protocol. To understand the dynamics of the heavy chain and the effect of light chain pairing on it, we firstly tried to obtain an artificial light chain that pairs with a broad range of heavy chains and then analyzed its effect on the antigen binding of heavy chains upon pairing. During the pre-B cell stage, the surrogate light chain (SLC) could pair with the nascent immunoglobulin μ heavy chains (Ig-μH) and promote them to function in the periphery. On the basis of this property, we designed several versions of genetically engineered "common light chain" prototypes by modifying the SLC structure. Among them, the mouse-derived VpreB1λ5Cκ light chain showed acceptable matching property with several different heavy chains without losing specificity of the original heavy chains, though the antigen affinities were variable. The extent of matching depended on the heavy chain; surprisingly, a specific heavy chain (IGHV9-3) could match with two different conventional Vκs (IGKV3-2*01 and IGKV10-96*01) without losing the antigen affinities, whereas another heavy chain (IGHV1-72) completely lost its antigen affinities by the same matching. Thus, the results suggested that the antigen recognition of the heavy chain is variably affected by the paired light chain, and that the artificial light chain, Mm_VpreB1λ5Cκ, has the potential to be a "common light chain", providing a novel system to analyze the effects of light chains in antigen recognition of heavy chains.

While two memory compartments, memory B cells and long-lived plasma cells, are thought to contribute to the successful establishment of memory recall responses, the unique roles of each cellular compartment are still unclear. Herein, by tracing influenza anti-hemagglutinin (HA)-specific antibodies in mice, we demonstrate that pre-existing antibodies secreted by long-lived plasma cells are essential for protection from reinfection with the same influenza virus, whereas protection from secondary infection with an antigenically distinct influenza virus requires memory B-cell activation. These activated memory B cells were largely specific for the conserved HA stem region, and generated sufficient levels of antibodies for protection from heterologous reinfection. Given that the anti-stem plasmablasts derived from the memory B cells were higher affinity than those from naive B cells, our results suggest that maturation of anti-stem memory B cells during primary influenza infection and their subsequent activation are required for protection from reinfection by mutant viruses.

The immense adaptability of antigen recognition by antibodies is the basis of the acquired immune system. Despite our understanding of the molecular mechanisms underlying the production of the vast repertoire of antibodies by the acquired immune systems, it has not yet been possible to arrive at a global view of a complete antibody repertoire. In particular, B cell repertoires have been regarded as a black box because of their astronomical number of antibody clones. However, next-generation sequencing technologies are enabling breakthroughs to increase our understanding of the B cell repertoire. In this report, we describe a simple and efficient method to visualize and analyze whole individual mouse and human antibody repertoires. From the immune organs, representatively from spleen in mice and peripheral blood mononuclear cells in humans, total RNA was prepared, reverse transcribed, and amplified using the 5'-RACE method. Using a universal forward primer and antisense primers for the antibody class-specific constant domains, antibody mRNAs were uniformly amplified in proportions reflecting their frequencies in the antibody populations. The amplicons were sequenced by next-generation sequencing (NGS), yielding more than 105 antibody sequences per immunological sample. We describe the protocols for antibody sequence analyses including V(D)J-gene-segment annotation, a bird's-eye view of the antibody repertoire, and our computational methods.

The immunogenicities of inactivated whole and split virus vaccines derived from influenza A/H1N1pdm09 virus were compared in a mouse model. We demonstrated the unique properties of whole virus vaccine boosters on the serum memory antibody response in mice. Consistent with previous studies, booster immunization with either whole or split virus vaccines of A/H1N1pdm09 virus produced comparable titers of serum antibodies with hemagglutination inhibition and virus-neutralizing activities. However, superior protection against the challenge infection was unexpectedly observed in mice primed and boosted with whole virus vaccines compared with those treated with split virus vaccines, despite similar levels of antibody titers in each group. Immune serum antibodies were shown to be primarily responsible for this protection via passive transfer experiments of immune serum antibodies to naive recipient mice. Moreover, this protection correlated with elevated affinity maturation of the antibodies. Thus, booster immunization with whole virus vaccines elicited a robust serum antibody response with high avidity to the virus, which was not measurable via conventional serological assays.

Abaloparatide (ABL) is a novel 34-amino acid peptide analog of parathyroid hormone-related protein. In clinical studies, although ABL showed a greater bone mineral density (BMD) increase than teriparatide (TPTD, human parathyroid hormone 1–34), the responses of ABL to bone formation and resorption markers were weaker, making it difficult to understand the relationship between the bone anabolic window (increase in bone formation versus resorption) and bone mass. In the present study, the effects of ABL and TPTD were compared in mice. Given that the rate of bone turnover is higher in rodents than in humans, the comparison was made with several administration regimens providing equivalent daily dosages: once daily (QD, 30 μg/kg every 24 h), twice daily (BID, 15 μg/kg every 12 h), or three times a day (TID, 10 μg/kg every 8 h). Frequent administration of ABL showed higher BMD with enhancement of trabecular and cortical bone mass and structures than that of TPTD, consistent with the clinical results seen with once daily administration. ABL increased bone formation marker levels more than TPTD with more frequent regimens, while bone resorption marker levels were not different between ABL and TPTD in all regimens. Analysis of bone histomorphometry and gene expression also suggested that ABL increased bone formation more than TPTD, while the effect on bone resorption was almost comparable between ABL and TPTD. The bone anabolic windows calculated from bone turnover markers indicated that ABL enhanced the anabolic windows more than TPTD, leading to a robust increase in BMD. The mechanism by which ABL showed a better balance of bone turnover was suggested to be partly due to the enhanced remodeling-based bone formation involved in Ephb4. Taken together, our findings would help elucidate the mechanism by which ABL shows excellent BMD gain and reduction of fractures in patients with osteoporosis.

Virus infection induces the development of T follicular helper (TFH) and T helper 1 (TH1) cells. Although TFH cells are important in anti-viral humoral immunity, the contribution of TH1 cells to a protective antibody response remains unknown. We found that IgG2 antibodies predominated in the response to vaccination with inactivated influenza A virus (IAV) and were responsible for protective immunity to lethal challenge with pathogenic H5N1 and pandemic H1N1 IAV strains, even in mice that lacked TFH cells and germinal centers. The cytokines interleukin-21 and interferon-γ, which are secreted from TH1 cells, were essential for the observed greater persistence and higher titers of IgG2 protective antibodies. Our results suggest that TH1 induction could be a promising strategy for producing effective neutralizing antibodies against emerging influenza viruses.

Despite the importance of memory B cells in protection from reinfection, how such memory cells are selected and generated during germinal-center (GC) reactions remains unclear. We found here that light-zone (LZ) GC B cells with B cell antigen receptors (BCRs) of lower affinity were prone to enter the memory B cell pool. Mechanistically, cells in this memory-prone fraction had higher expression of the transcriptional repressor Bach2 than that of their counterparts with BCRs of higher affinity. Haploinsufficiency of Bach2 resulted in reduced generation of memory B cells, independently of suppression of the gene encoding the transcription factor Blimp-1. Bach2 expression in GC cells was inversely correlated with the strength of help provided by T cells. Thus, we propose an instructive model in which weak help from T cells maintains relatively high expression of Bach2, which predisposes GC cells to enter the memory pool.

Symptomatic epidural haematoma after cervical laminoplasty is rare. We report 3 patients who required emergency evacuation of an epidural haematoma. Timely diagnosis and removal of the haematoma is important to prevent neurological deficits. The causative factors in these patients were preoperative coagulopathy, hypertension, and the malfunction of a closed-suction drain.

In order to directly investigate the grain boundary (GB) hydrogen embrittlement in polycrystalline materials, a novel micro-mechanical testing method was developed. By combining a site-specific sampling technique and a high-voltage environmental transmission electron microscope (HV ETEM), the fracture property of micro-cantilever specimens fabricated from the same GB in a nickel-aluminide (Ni 3 Al) polycrystal was critically compared in environments with/without hydrogen (H 2 ) gas. For randomly oriented GBs, brittle fracture nucleation accompanied by plastic deformation was observed in a H 2 -containing environment except for ones with small orientation difference. No GB fracture was observed for coherent Σ3 boundaries. It also appeared that the similitude of the hydrogen-enhanced decohesion (HEDE) mechanism was still valid even for the submicron-scale specimens.

Computational drug design plays an important role in the discovery of new drugs. Recently, we proposed an algorithm for designing new drug-like molecules utilizing the structure of a known active molecule. To design molecules, three types of fragments (ring, linker, and side-chain fragments) were defined as building blocks, and a fragment library was prepared from molecules listed in G protein-coupled receptor (GPCR)-SARfari database. An evolutionary algorithm which executes evolutionary operations, such as crossover, mutation, and selection, was implemented to evolve the molecules. As a case study, some GPCRs were selected for computational experiments in which we tried to design ligands from simple seed fragments using the Tanimoto coefficient as a fitness function. The results showed that the algorithm could be used successfully to design new molecules with structural similarity, scaffold variety, and chemical validity. In addition, a docking study revealed that these designed molecules also exhibited shape complementarity with the binding site of the target protein. Therefore, this is expected to become a powerful tool for designing new drug-like molecules in drug discovery projects.

In order to clarify the plastic deformation mechanism of silicon carbide in cubic phase (3C-SiC), molecular dynamics (MD) simulations are performed on the nanoindentation using a spherical indenter. Transition from elastic deformation to plastic deformation has been confirmed by the phenomenon called pop-in in the load-displacement curves during nanoindentation. Dislocations on {1 1 1} slip planes are found during indentation. In order to analyze internal defects, common neighbor analysis (CNA) is slightly modified so that it is suitable for the analysis of slips of zinc-blend structure. In our method, the CNA is applied separately to sub-lattice of Si or C in the same SiC. By this method, structural changes are confirmed in a region with the shape of square pyramid when the pop-in behavior occurs. By measuring the atomic distances along the region of misalignment, it was confirmed that there is certainly atomic sliding by crystalline slip. Furthermore, it is found that, with increase of loading, dislocation loops spread along {1 1 1} slip planes.

A new environmental high-voltage transmission electron microscope (E-HVEM) was developed by Nagoya University in collaboration with JEOL Ltd. An open-type environmental cell was employed to enable in-situ observations of chemical reactions on catalyst particles as well as mechanical deformation in gaseous conditions. One of the reasons for success was the application of high-voltage transmission electron microscopy to environmental (in-situ) observations in the gas atmosphere because of high transmission of electrons through gas layers and thick samples. Knock-on damages to samples by high-energy electrons were carefully considered. In this paper, we describe the detailed design of the E-HVEM, recent developments and various applications. This article is part of a discussion meeting issue ‘Dynamic in situ microscopy relating structure and function'.

The microscopic mechanism of high temperature (900 °C) fatigue crack growth in a single crystal Ni-based superalloy is investigated. Particular attention is focused to its relation to the cyclic misorientation markings (CMMs), formerly termed EBSD (electron backscattered diffraction) striation, left along the crack wake. Observation of a cross-sectioned crack tip is conducted by various electron microscopy i.e. SEM, EBSD, high-voltage TEM and STEM-EDS. A {100} fatigue crack subjected to a relatively high ΔK (40 MPam1/2) shows CMMs with an interval corresponding to crack growth rate (2–4 μm/cycle), consisting of heavily dislocated γ (Ni) phase and less dislocated γ’ (Ni3Al) phase. It is also revealed that local oxidation along the γ/γ’ interface at the crack tip existed at the current temperature/ΔK level. On the bases of these findings, a rational yet qualitative crack growth model that incorporated both brittle/ductile processes is proposed.

In this study, a conventional precipitation-hardened A6061-T6 alloy (base alloy) and a modified alloy containing excess solute magnesium (Mg) compared to the base alloy were compared in terms of high-cycle fatigue property. Particular attention was paid to the emergence of a distinct knee point on the fatigue life diagram together with a time-dependent strengthening (coaxing) effect in the modified alloy under various test conditions. An attempt using specimens with different defect size successfully revealed that a distinct knee, which was absent in the base alloy, was attributed to the threshold against small crack growth. The clear coaxing effect in the modified alloy was also explicitly confirmed in terms of the high ΔK value achieved by the arrested cracks regardless of the defect size. The comparative tests conducted under different environments and temperature conditions further confirmed that the crack tip strengthening, essentially caused by the strain-aging capability endowed to the modified alloy, operated most prominently in the ambient air at room temperature. Such results were rationally interpreted by incorporating a competitive interaction of solute Mg and hydrogen (H) with dislocations at the crack tip.

Phase-separated glass (PSG) is being used widely for manufacturing scientific apparatus and kitchenware, owing to its advantages of high hardness and heat resistance, which can be attributed to its sea- and island-like structures on the nanoscale. There is a growing demand in the glass industry for methods for evaluating and visualizing the local stress distribution in such glass materials at the nanoscale in order to elucidate their fracture mechanism and to obtain more information to develop even harder glass materials. In this study, we clarified the correlation between the local stress in this type of glass material and its associated nanoscale structure using a nanoelectron probe based on scanning transmission electron microscopy and cathodoluminescence. The peak shift of an emission line corresponding to an intentionally doped rare-earth element was measured based on the piezospectroscopic effect. We found that the sea- and island-like structures, whose origins can be attributed to the difference in the thermal expansion coefficients of the two phases, mutually push each other. This result was confirmed by a simple finite-element method simulation. The present result suggests that the local mechanical properties of PSG can be well described by a coherent elastic continuum consisting of the phases that evolve during thermal treatment.