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Abstract Background We previously demonstrated that the hydroxymethylglutaryl-CoA (HMG-CoA) reductase inhibitor (statins) play an important role in the regulation of alloimmune responses. However, little is known regarding the effects of statin on allograft protection or donor-specific antibodies (DSA). In this study, we investigated the graft-protective and immunomodulatory effects of rosuvastatin in a model of fully major histocompatibility complex-mismatched murine cardiac allograft transplantation. Methods CBA mice underwent transplantation of C57BL/6 (B6) hearts and received 50 and 500 μg/kg/day of rosuvastatin from the day of transplantation until seven days after the completion of transplantation. To confirm the requirement for regulatory T cells (Tregs), we administered an anti-interleukin-2 receptor alpha antibody (PC-61) to rosuvastatin-treated CBA recipients. Additionally, histological and fluorescent staining, cell proliferation analysis, flow cytometry, and DSA measurements were performed. Results CBA recipients with no treatment rejected B6 cardiac graft acutely (median survival time [MST], 7 days). CBA mice treated with 500 μg/kg/day of rosuvastatin prolonged allograft survival (MSTs, 77 days). Fluorescent staining studies showed that rosuvastatin-treated recipients had strong aggregation of CD4+Foxp3+ cells in the myocardium and around the coronary arteries of cardiac allografts two weeks after grafting. Flow cytometry studies performed two weeks after transplantation showed an increased number of splenic CD4+CD25+Foxp3+ T cells in rosuvastatin-treated recipients. The addition of rosuvastatin to mixed leukocyte cultures suppressed cell proliferation by increasing the number of CD4+CD25+Foxp3+ Tregs. Additionally, Tregs suppressed DSA production in rosuvastatin-treated recipients. Conclusion Rosuvastatin treatment may be a complementary graft-protective strategy for suppressing DSA production in the acute phase, driven by the promotion of splenic and graft-infiltrating CD4+CD25+Foxp3+ Tregs.

Abstract Allergic rhinitis (AR) is caused by type I hypersensitivity reaction in the nasal tissues. The interaction between CD300f and its ligand ceramide suppresses immunoglobulin E (IgE)-mediated mast cell activation. However, whether CD300f inhibits the development of allergic rhinitis (AR) remains elusive. We aimed to investigate the roles of CD300f in the development of AR and the effectiveness of intranasal administration of ceramide liposomes on AR in murine models. We used ragweed pollen-induced AR models in mice. Notably, CD300f deficiency did not significantly influence the ragweed-specific IgE production, but increased the frequency of mast cell-dependent sneezing as well as the numbers of degranulated mast cells and eosinophils in the nasal tissues in our models. Similar results were also obtained for MCPT5-exprssing mast cell-specific loss of CD300f. Importantly, intranasal administration of ceramide liposomes reduced the frequency of sneezing as well as the numbers of degranulated mast cells and eosinophils in the nasal tissues in AR models. Thus, CD300f–ceramide interaction, predominantly in mast cells, alleviates the symptoms and progression of AR. Therefore, intranasal administration of ceramide liposomes may be a promising therapeutic approach against AR by targeting CD300f.

Background: Multidisciplinary efforts to prospectively collect and analyze symptoms of hay fever are limited. We aimed to identify the characteristics of nasal and ocular symptoms of hay fever, using the AllerSearch smartphone application. Methods: This mobile health-based prospective observational study using the AllerSearch smartphone application was conducted between February 1, 2018, and May 1, 2020. Individuals who downloaded AllerSearch from Japan and provided comprehensive self-assessments (including 17 items related to quality of life [QoL]-related items) were included. The characteristics and risk factors for allergic rhinitis (AR) and allergic conjunctivitis (AC) were identified using hierarchical heat maps and multivariate logistic regression. Results: Of the 9041 participants with hay fever, 58.8% had AR and AC, 22.2% had AR, and 5.7% had AC. The AR–AC comorbid cohort showed worse symptoms of hay fever and QoL scores than the other cohorts. Factors (odds ratio, 95% confidence interval) associated with AR–AC included a lower age (0.98, 0.97–0.98), female sex (1.31, 1.19–1.45), liver disease (1.58, 1.26–2.35), dry eye disease (1.45, 1.30–1.63), unknown dry eye disease status (1.46, 1.31–1.62), contact lens use discontinuation during the hay fever season (1.69, 1.28–2.23), and bedroom flooring material other than hardwood, carpet, tatami, or vinyl (1.91, 1.16–3.14). Conclusions: Analysis of medical big data for hay fever performed using a mobile health app helped identify risk factors and characteristics of AC, AR, and AR–AC. Phenotyping of highly variable symptoms of hay fever, such as nasal and ocular symptoms, can facilitate better-quality clinical care.

In the pyro-reprocessing of spent nuclear fuel, salt bath is normally used in several times, but at final moment, spent salt containing small amount of chloride nuclear fuel material is generated. In terms of managing nuclear fuel materials, it is desirable that the nuclear fuel materials should be recovered from the spent salt. We are proposing methods for recovering nuclear fuel materials using precipitation by oxide addition and distillation for reducing pressure techniques. In this study, we have focused on the behavior of manganese(II), which is one of the radioactivated products. As a result of experiments and thermodynamic simulation, it was found that manganese(II) is likely to be entrained in nuclear fuel materials. Therefore, it is necessary to add a step to separate manganese(II) from nuclear fuel materials.

BackgroundDry eye (DE) and hay fever (HF) show synergistic exacerbation of each other’s pathology through inflammatory pathways. ObjectiveThis study aimed to investigate the association between DE and HF comorbidity and the related risk factors. MethodsA cross-sectional observational study was conducted using crowdsourced multidimensional data from individuals who downloaded the AllerSearch smartphone app in Japan between February 2018 and May 2020. AllerSearch collected the demographics, medical history, lifestyle and residential information, HF status, DE symptoms, and HF-related quality of life. HF symptoms were evaluated using the nasal symptom score (0-15 points) and nonnasal symptom score (0-12 points). HF was defined by the participants’ responses to the questionnaire as HF, non-HF, or unknown. Symptomatic DE was defined as an Ocular Surface Disease Index total score (0-100 points), with a threshold score of 13 points. HF-related quality of life was assessed using the Japanese Allergic Conjunctival Disease Standard Quality of Life Questionnaire (0-68 points). We conducted a multivariable linear regression analysis to examine the association between the severity of DE and HF symptoms. We subsequently conducted a multivariable logistic regression analysis to identify the factors associated with symptomatic DE (vs nonsymptomatic DE) among individuals with HF. Dimension reduction via Uniform Manifold Approximation and Projection stratified the comorbid DE and HF symptoms. The symptom profiles in each cluster were identified using hierarchical heat maps. ResultsThis study included 11,284 participants, classified into experiencing HF (9041 participants), non-HF (720 participants), and unknown (1523 participants) groups. The prevalence of symptomatic DE among individuals with HF was 49.99% (4429/9041). Severe DE symptoms were significantly associated with severe HF symptoms: coefficient 1.33 (95% CI 1.10-1.57; P

Plain English summary Patient and public involvement (PPI) plays an important role in promoting effective execution of health science research, as well as in the establishment of a social agreement and infrastructure for the care of various diseases, including cancer, chronic diseases, and allergic illnesses. Hay fever is one of the most common allergic diseases, affecting more than 30 million people in Japan. It is known for its myriad factors and diverse presentations. Previously, we developed a mobile health (mHealth) smartphone application (app) for hay fever—AllerSearch—released in February 2018. This app is capable of collecting relevant digital phenotypes and user-provided information, which are used in providing tailored, evidence-based suggestions. To our knowledge, no other studies have been conducted on the implementation of PPI in mHealth. Since hay fever presents with a wide variety of symptoms and risk factors, PPI principles appear well-suited for eliciting insights from the patient/public population and for incorporating new, expert perspectives into the research process. In this study, we included PPI contributors in the research plan, app development, and evaluation. Most notably, the survey questionnaire and user interface of the app was tailored based on PPI feedback. The updated AllerSearch app was released during this study period. Since hay fever is a widespread and variable illness, the multifaceted input from patients and public experts enabled by PPI implementation holds promise for improving society-wide healthcare and in empowering a culture toward medical involvement.

Background: The prevalence of hay fever, a multifactorial allergic disease, is increasing. Identifying individual characteristics and associated factors of hay fever is essential for predictive, preventive, personalized, and participatory (P4) medicine. This study aimed to identify individual characteristics and associated factors of hay fever using an iPhone application AllerSearch. Methods: This large-scale mobile health-based cross-sectional study was conducted between February 2018 and May 2020. Individuals who downloaded AllerSearch in Japan and provided a comprehensive self-assessment (general characteristics, medical history, lifestyle habits, and hay fever symptoms [score range 0–36]) were included. Associated factors of hay fever (vs. non-hay fever) and severe hay fever symptoms were identified using multivariate logistic and linear regression analyses, respectively. Results: Of the included 11,284 individuals, 9041 had hay fever. Factors associated with hay fever (odds ratio) included age (0.98), female sex (1.33), atopic dermatitis (1.40), history of dry eye diagnosis (1.36), discontinuation of contact lens use during hay fever season (3.34), frequent bowel movements (1.03), and less sleep duration (0.91). The factors associated with severe hay fever symptoms among individuals with hay fever (coefficient) included age (−0.104), female sex (1.329), history of respiratory disease (1.539), history of dry eye diagnosis (0.824), tomato allergy (1.346), discontinuation of contact lens use during hay fever season (1.479), smoking habit (0.614), and having a pet (0.303). Conclusions: Our large-scale mobile health-based study using AllerSearch elucidated distinct hay fever presentation patterns, characteristics, and factors associated with hay fever. Our study establishes the groundwork for effective individualized interventions for P4 medicine.

Abstract Preservation is critical for egg and larval samples from ichthyoplankton surveys. As growing interest is directed toward maternal effects on egg quality, historical collections of egg samples in formalin are potentially invaluable materials but concerns about the effects of formalin preservation need to be clarified. The present study examined effects of formalin preservation on egg size of Japanese sardine Sardinops melanostictus, chub mackerel Scomber japonicus and Japanese anchovy Engraulis japonicus over a long period (30–36 months). Overall, the egg sizes slightly decreased in a couple of months after preservation but then recovered to be at a similar level to the live eggs for diameters of spherical eggs of sardine and mackerel and short diameters of ellipsoidal eggs of anchovy, although a caveat lies in unstable and slightly increasing patterns in long diameters of anchovy eggs. No substantial differences were found between 5% and 10% formalin solutions. For practical purposes, we conclude that no correction factors are required for using the measurements of egg samples preserved in formalin over a long period. Assuming the applicability to congeneric species, the information presented here supports the validity of previous studies and will promote future studies on egg size variability in small pelagic fish based on historical collections of egg samples preserved in formalin in various regions of the world.

Abstract Dyslipidemia is considered an essential component of the pathological process of amyotrophic lateral sclerosis (ALS), a fatal motor neuron disease. Although TAR DNA Binding Protein 43 kDa (TDP-43) links both familial and sporadic forms of ALS and cytoplasmic aggregates are a hallmark of most cases of ALS, the molecular mechanism and the in vivo relation of ALS dyslipidemia with TDP-43 have been unclear. To analyze the dyslipidemia-related gene expression by TDP-43, we performed expression microarray and RNA deep sequencing (RNA-Seq) using cell lines expressing high levels of TDP-43 and identified 434 significantly altered genes including sterol regulatory element-binding protein 2 (SREBP2), a master regulator of cholesterol homeostasis and its downstream genes. Elevated TDP-43 impaired SREBP2 transcriptional activity, leading to inhibition of cholesterol biosynthesis. The amount of cholesterol was significantly decreased in the spinal cords of TDP-43-overexpressed ALS model mice and in the cerebrospinal fluids of ALS patients. These results suggested that TDP-43 could play an essential role in cholesterol biosynthesis in relation to ALS dyslipidemia.

Abstract We investigated the effects of anti-CD80/86 antibodies in a murine high-risk corneal transplantation rejection model. A mixed lymphocyte reaction (MLR) assay was conducted with anti-CD80/86 antibodies. Inflammatory cytokine levels in the culture supernatant were measured using an enzyme-linked immunosorbent assay. Interferon (IFN)-γ-producing CD4+ T cell frequencies in the MLR were assessed using flow cytometry. In vivo, high-risk corneal allograft survival and IFN-γ-producing CD4+ T cell frequencies in corneal grafts were assessed with intraperitoneal injection of anti-CD80/86 antibodies compared to phosphate-buffered saline (PBS). RNA-sequencing was performed on corneal grafts 2 weeks post-transplantation. Anti-CD80/86 antibodies significantly decreased T-cell proliferation, IFN-γ+-producing CD4+ T cell frequencies, and IFN-γ, interleukin (IL)-1β, IL-2, IL-10, and tumor necrosis factor-α production in the MLR compared to PBS injection. Intraperitoneal injection of anti-CD80/86 antibodies significantly prolonged corneal graft survival and decreased IFN-γ+-producing CD4+ T cell frequencies compared to PBS injection. Gene set enrichment analysis showed that the gene sets mainly enriched in the control group were related to allograft rejection and inflammatory response compared to PBS injection. Anti-CD80/86 antibodies significantly prolonged corneal graft survival by inhibiting T-cell proliferation and inflammatory response.

Abstract Multidimensional integrative data analysis of digital phenotyping is crucial for elucidating the pathologies of multifactorial and heterogeneous diseases, such as the dry eye (DE). This crowdsourced cross-sectional study explored a novel smartphone-based digital phenotyping strategy to stratify and visualize the heterogenous DE symptoms into distinct subgroups. Multidimensional integrative data were collected from 3,593 participants between November 2016 and September 2019. Dimension reduction via Uniform Manifold Approximation and Projection stratified the collected data into seven clusters of symptomatic DE. Symptom profiles and risk factors in each cluster were identified by hierarchical heatmaps and multivariate logistic regressions. Stratified DE subgroups were visualized by chord diagrams, co-occurrence networks, and Circos plot analyses to improve interpretability. Maximum blink interval was reduced in clusters 1, 2, and 5 compared to non-symptomatic DE. Clusters 1 and 5 had severe DE symptoms. A data-driven multidimensional analysis with digital phenotyping may establish predictive, preventive, personalized, and participatory medicine.

Maternal effects on offspring viability significantly contribute to the productivity and resilience of exploited marine fish populations. The temporal trend of biomass of chub mackerel (Scomber japonicus) in the western North Pacific showed cyclic fluctuation patterns, but it has been rapidly increasing since 2011. Although there are several assumptions regarding the mechanisms of the population dynamics of this species in these waters, there is a contradiction in the effects of thermal environmental conditions on recruitment success. We explored the effects of maternal age and/or previous spawning experience on egg and offspring traits of captive chub mackerel in response to thermal conditions. Moreover, from the relationship between the growth rate and temperature experienced by captive fish, the experienced temperatures of wild specimens were assessed. The sizes of eggs spawned from females aged 1–3 years were negatively correlated with temperature, but the effects of maternal age on egg size and nutrition were significant with larger and nutrient-rich eggs spawned from 2- or 3-year-old females (repeat spawners) than those from 1-year-old females (first-time spawners) at a given temperature. Such temperature- and maternal-age-dependent egg traits reflected the standard length of first-feeding larvae. The effects of maternal age on the starvation tolerance of larvae after hatching and body sizes of first-feeding larvae were also evident in larvae derived from 3-year-old females showing longer starvation tolerance and larger body heights than those from 1-year-old females. Further, larvae derived from 3-year-old females grew significantly faster than those from 1-year-old females at the tested temperatures. The process of ontogenetic development and growth rate of captive specimens until age 25 days were positively correlated with temperature. Meanwhile, the estimated temperatures experienced by wild specimens until age 25 days ranged from 18.0 to 20.3°C. These findings suggested that there were opposing thermal responses between maternal and offspring traits of chub mackerel, but the temperatures experienced by the main recruit fish may be restricted within the range of temperatures available for spawning. We discuss the mechanisms of reproductive success of this species in accordance with the maternal spawning experience and thermal effects on life history traits.

BackgroundHay fever is a highly prevalent, heterogenous, and multifactorial disease. Patients may benefit from longitudinal assessments using mobile health (mHealth) principles. We have previously attempted to establish an effective mHealth platform for patients with hay fever through AllerSearch, our in-house smartphone app that assesses electronic patient-reported outcomes through a questionnaire on hay fever and provides evidence-based advice. To be used by the public, an investigation on its reliability and validity is necessary. ObjectiveThe aim of this paper is to assess the reliability and validity of subjective symptom data on hay fever collected through our app, AllerSearch. MethodsThis study used a prospective observational design. The participants were patients aged ≥20 years recruited from a single university hospital between June 2, 2021, and January 26, 2022. We excluded patients who could not use smartphones as well as those with incomplete data records and outlier data. All participants answered the Japanese Allergic Conjunctival Disease Standard Quality of Life Questionnaire (JACQLQ), first in the paper-and-pencil format and subsequently on AllerSearch on the same day. The JACQLQ comprises the following three domains: Domain I, with 9 items on ocular or nasal symptoms; Domain II, with 17 items on daily activity and psychological well-being; and Domain III, with 3 items on overall condition by face score. The concordance rate of each domain between the 2 platforms was calculated. The internal consistency of Domains I and II of the 2 platforms was assessed using Cronbach alpha coefficients, the concurrent validity of Domains I and II was assessed by calculating Pearson correlation coefficients, and the mean differences between the 2 platforms were assessed using Bland-Altman analysis. ResultsIn total, 22 participants were recruited; the data of 20 (91%) participants were analyzed. The average age was 65.4 (SD 12.8) years, and 80% (16/20) of the participants were women. The concordance rate of Domains I, II, and III between the paper-based and app-based JACQLQ was 0.78, 0.85, and 0.90, respectively. The internal consistency of Domains I and II between the 2 platforms was satisfactory (Cronbach alpha of .964 and .919, respectively). Pearson correlation analysis yielded a significant positive correlation between Domains I and II across the 2 platforms (r=0.920 and r=0.968, respectively). The mean difference in Domains I and II between the 2 platforms was 3.35 units (95% limits of agreement: –6.51 to 13.2). ConclusionsOur findings indicate that AllerSearch is a valid and reliable tool for the collection of electronic patient-reported outcomes to assess hay fever, contributing to the advantages of the mHealth platform.

Summary: Thermogenic brown and beige adipocytes counteract obesity by enhancing energy dissipation via uncoupling protein-1 (Ucp1). However, the effect of genetic variation on these cells, a major source of disease susceptibility, has been less well studied. Here we examined beige adipocytes from obesity-prone C57BL/6J (B6) and obesity-resistant 129X1/SvJ (129) mouse strains and identified a cis-regulatory variant rs47238345 that is responsible for differential Ucp1 expression. The alternative T allele of rs47238345 at the Ucp1 -12kb enhancer in 129 facilitates the allele-specific binding of nuclear factor I-A (NFIA) to mediate allele-specific enhancer-promoter interaction and Ucp1 transcription. Furthermore, CRISPR-Cas9/Cpf1-mediated single nucleotide polymorphism (SNP) editing of rs47238345 resulted in increased Ucp1 expression. We also identified Lim homeobox protein 8 (Lhx8), whose expression is higher in 129 than in B6, as a trans-acting regulator of Ucp1 in mice and humans. These results demonstrate the cis- and trans-acting effects of genetic variation on Ucp1 expression that underlie phenotypic diversity.

In this paper, we develop distributed algorithms for achieving resilient consensus via the maximum value-based approach when adversarial agents may be present in the network. The adversaries intend to prevent the nonfaulty, normal agents from reaching consensus. We extend the class of resilient methods known as the mean subsequence reduced (MSR) algorithms, where the agents make selections on their neighbours' information at the time of their updates so as to reduce the influence of the malicious agents. In particular, the normal agents try to arrive at the maximum value of their initial states. Due to the malicious agents, the exact maximum may not be reached, the advantage of the approach is the finite-time convergence. We present both synchronous and asynchronous versions of the update schemes and characterize graph theoretic conditions for achieving resilient consensus. A numerical example is provided to illustrate our results.

Allergic conjunctivitis affects approximately 15–20% of the global population and can permanently deteriorate one's quality of life (QoL) and work productivity, leading to societal work force costs. Although not fully understood, allergic conjunctivitis is a multifactorial disease with a complex network of environmental, lifestyle, and host contributory risk factors. To effectively enhance the quality of treatment for patients with allergic conjunctivitis, as well as other allergic diseases, the field must first comprehend the pathology underlying various individualized subjective symptoms and stratify the disease according to risk factors and presentations. Such competent stratification and societal reconstruction that targets the alleviation of the damage due to allergic diseases would greatly help ramify personalized treatments and prevent the projected increase in societal costs imposed by allergic diseases.Owing to the rapid advancements in the information and technology sector, medical big data are greatly accessible and useful to decipher the pathophysiology of many diseases. Such data collected through multi-omics and mobile health have been effective for research on chronic diseases including allergic and immune-mediated diseases. Novel big data containing vast and continuous information on individuals with allergic conjunctivitis and other allergic symptoms are being used to search for causative genes of diseases, gain insights into new biomarkers, prevent disease progression, and, ultimately, improve QoL. The individualized and holistic data accrued from new angles using technological innovations are helping the field realize the principles of P4 medicine: predictive, preventive, personalized, and participatory medicine.

Abstract Background The clupeoid fishes are ecologically and commercially important fish species worldwide that exhibit a high level of population fluctuation, accompanied by alteration of reproductive traits. However, knowledge about their reproductive physiology in order to understand mechanisms underlying such population dynamics is limited. The endocrine system along with the brain–pituitary–gonadal (BPG) axis is critical for regulating reproduction. The aims of this study were to provide transcript data and genes related to the BPG axis, and to characterize the expression profiles of ovarian steroidogenesis-related genes in the Japanese sardine (Sardinops melanostictus, Clupeidae). Results RNA sequencing was performed using the sardine brain, pituitary, and gonad in both sexes. A total of 290,119 contigs were obtained and 115,173 non-redundant ORFs were annotated. The genes differentially expressed between ovary and testis were strongly associated with GO terms related to gamete production. The tissue-specific profile of the abundance of transcripts was characterized for the major regulators in the BPG axis, such as gonadotropin-releasing hormone, gonadotropin, and steroidogenic enzyme. By comparing between ovary and testis, out of eight different 17β-hydroxysteroid dehydrogenase (Hsd17b) genes identified, higher hsd17b7 expression was found in testis, whereas higher expression of hsd17b8, hsd17b10, hsd17b12a, and hsd17b12b was found in ovary. The cDNAs encoding key endocrine factors in the ovarian steroidogenic pathway were cloned, sequenced, and quantitatively assayed. In the pituitary, follicle-stimulating hormone beta peaked during vitellogenesis, while luteinizing hormone beta peaked at the completion of vitellogenesis. In the ovary, follicle-stimulating hormone receptor and luteinizing hormone receptor were upregulated from mid- to late phase of vitellogenesis. Furthermore, three steroidogenic enzyme genes (cyp11a1, cyp17a1, and cyp19a1a) gradually increased their expression during ovarian development, accompanying a rise in serum estradiol-17β, while 3β-hydroxysteroid dehydrogenase and steroidogenic acute regulatory protein did not change significantly. Conclusions This is the first report of deep RNA sequencing analysis of Japanese sardine, in which many key genes involved in the BPG axis were identified. Expression profiles of ovarian steroidogenesis-related genes provide a molecular basis of the physiological processes underlying ovarian development in the sardine. Our study will be a valuable resource for clarifying the molecular biology of clupeoid fishes.

Given the time scale based on the duration of exposure to global warming, natural climate-gradient studies and experimental manipulations have detected long-term (decades to centuries) and short-term (years to decades) ecological responses to global warming. Combination of these two complementary approaches within a single study may enable prediction of the likely responses of ecological processes to global warming. To understand how global warming affects plant–herbivore interactions within a canopy of Erman’s birch, we combined an elevational gradient study and a warming experiment involving mature birch trees in which the soil and tree branches were warmed separately. In the elevational gradient study, herbivory by chewing insects and plant growth increased as elevation decreased, and the concentrations of condensed tannins and total phenolics in the leaves decreased. In the warming experiment, soil warming alone increased herbivory, and the addition of branch warming amplified the effect on herbivory. Soil warming alone decreased the tannin concentration, and the addition of branch warming led to a further reduction. The variation in herbivory was best explained by the tannin content of leaves. Our experimental results demonstrate that the decreased tannin content of leaves due to a combination of soil and branch warming was an important driver of increased herbivory in the canopy of the mature birch trees. The similar tendencies in the short- and long-term responses imply that global warming is likely to increase background herbivory in mature birch trees by decreasing the tannin content of leaves in the canopy.

Species-specific ecological traits in fishes are likely to vary between populations or stocks due to differences in regional oceanic conditions, such as latitudinal temperature. We examined potential intraspecific differences in the swimming performance and metabolism of Pacific chub mackerel (Scomber japonicus) from the Northwest and Northeast Pacific stocks, which are distributed on opposite sides of the North Pacific at similar latitudes, but where the temperature contrast is large. Swimming bioenergetics and metabolic data of Northwest stock mackerel were measured at 14, 18, and 24°C using variable-speed swim-tunnel respirometers, and then the resulting bioenergetic parameters were compared with previous findings from the Northeast stock. At a given size, the maximum sustainable swimming speed (Umax) of the Northwest stock showed no significant difference compared to the Northeast stock at 18 and 24°C, but was lower at 14°C. In addition, the oxygen consumption rate (MO2) of the Northwest stock showed lower mass dependence and different temperature dependence at a given swimming speed than in the Northeast stock. Combined with stock-specific data on growth and experienced temperatures in the wild, these bioenergetic differences indicate that the swimming performance and metabolism of the two stocks are specific to their local environment to maximize bioenergetic efficiency.

Paired related homeobox 1 (PRRX1) is a marker of limb bud mesenchymal cells, and deficiency of p53 or Rb in Prrx1-positive cells induces osteosarcoma in several mouse models. However, the regulatory roles of PRRX1 in human osteosarcoma have not been defined. In this study, we performed PRRX1 immunostaining on 35 human osteosarcoma specimens to assess the correlation between PRRX1 level and overall survival. In patients with osteosarcoma, the expression level of PRRX1 positively correlated with poor prognosis or the ratio of lung metastasis. Additionally, we found PRRX1 expression on in 143B cells, a human osteosarcoma line with a high metastatic capacity. Downregulation of PRRX1 not only suppressed proliferation and invasion but also increased the sensitivity to cisplatin and doxorubicin. When 143B cells were subcutaneously transplanted into nude mice, PRRX1 knockdown decreased tumor sizes and rates of lung metastasis. Interestingly, forskolin, a chemical compound identified by Connectivity Map analysis using RNA expression signatures during PRRX1 knockdown, decreased tumor proliferation and cell migration to the same degree as PRRX1 knockdown. These results demonstrate that PRRX1 promotes tumor malignancy in human osteosarcoma.

Stem cell-based therapies and experimental methods rely on efficient induction of human pluripotent stem cells (hPSCs). During limb development, the lateral plate mesoderm (LPM) produces limb-bud mesenchymal (LBM) cells that differentiate into osteochondroprogenitor cells and form cartilage tissues in the appendicular skeleton. Previously, we generated PRRX1-tdTomato reporter hPSCs to establish the protocol for inducing the hPSC-derived PRRX1+ LBM-like cells. However, surface antigens that assess the induction efficiency of hPSC-derived PRRX1+ LBM-like cells from LPM have not been identified. Here, we used PRRX1-tdTomato reporter hPSCs and found that high pluripotent cell density suppressed the expression of PRRX1 mRNA and tdTomato after LBM-like induction. RNA sequencing and flow cytometry suggested that PRRX1-tdTomato+ LBM-like cells are defined as CD44high CD140Bhigh CD49f−. Importantly, other hPSC lines, including four human induced pluripotent stem cell lines (414C2, 1383D2, HPS1042, HPS1043) and two human embryonic stem cell lines (SEES4, SEES7), showed the same results. Thus, an appropriate cell density of hPSCs before differentiation is a prerequisite for inducing the CD44high CD140Bhigh CD49f− PRRX1+ LBM-like cells.

Summary: Here, we report that MYC rescues early human cells undergoing reprogramming from a proliferation pause induced by OCT3/4, SOX2, and KLF4 (OSK). We identified ESRG as a marker of early reprogramming cells that is expressed as early as day 3 after OSK induction. On day 4, ESRG positive (+) cells converted to a TRA-1-60 (+) intermediate state. These early ESRG (+) or TRA-1-60 (+) cells showed a proliferation pause due to increased p16INK4A and p21 and decreased endogenous MYC caused by OSK. Exogenous MYC did not enhance the appearance of initial reprogramming cells but instead reactivated their proliferation and improved reprogramming efficiency. MYC increased expression of LIN41, which potently suppressed p21 post-transcriptionally. MYC suppressed p16 INK4A. These changes inactivated retinoblastoma protein (RB) and reactivated proliferation. The RB-regulated proliferation pause does not occur in immortalized fibroblasts, leading to high reprogramming efficiency even without exogenous MYC. : Rand et al. find that MYC promotes proliferation of human intermediate reprogrammed cells rather than initiation of reprogramming. MYC post-transcriptionally activates LIN41, resulting in post-transcriptional suppression of p21. Suppression of p21 results in reduction of RB activity, which is a negative regulator of reprogramming progression. Keywords: reprogramming, pluripotency, induced pluripotent stem cell, proliferation, senescence, immortalization, MYC, LIN41, post-transcriptional regulation

Abstract CDK5 Regulatory Subunit-Associated Protein 1-like 1 (CDKAL1) was identified as a susceptibility gene for type 2 diabetes and body mass index in genome-wide association studies. Although it was reported that CDKAL1 is a methylthiotransferase essential for tRNALys(UUU) and faithful translation of proinsulin generated in pancreatic β cells, the role of CDKAL1 in adipocytes has not been understood well. In this study, we found that CDKAL1 is expressed in adipose tissue and its expression is increased during differentiation. Stable overexpression of CDKAL1, however, inhibited adipocyte differentiation of 3T3-L1 cells, whereas knockdown of CDKAL1 promoted differentiation. CDKAL1 increased protein levels of β-catenin and its active unphosphorylated form in the nucleus, thereby promoting Wnt target gene expression, suggesting that CDKAL1 activated the Wnt/β-catenin pathway—a well-characterized inhibitory regulator of adipocyte differentiation. Mutant experiments show that conserved cysteine residues of Fe-S clusters of CDKAL1 are essential for its anti-adipogenic action. Our results identify CDKAL1 as novel negative regulator of adipocyte differentiation and provide insights into the link between CDKAL1 and metabolic diseases such as type 2 diabetes and obesity.

Summary: There has been increasing success with the generation of pancreatic cells from human induced pluripotent stem cells (hiPSCs); however, the molecular mechanisms of the differentiation remain elusive. The purpose of this study was to reveal novel molecular mechanisms for differentiation to PDX1+NKX6.1+ pancreatic endoderm cells, which are pancreatic committed progenitor cells. PDX1+ posterior foregut cells differentiated from hiPSCs failed to differentiate into pancreatic endoderm cells at low cell density, but Rho-associated kinase (ROCK) or non-muscle myosin II (NM II) inhibitors rescued the differentiation potential. Consistently, the expression of phosphorylated myosin light chain 2 and NM IIA was downregulated in aggregation culture. Notably, the soluble factors we tested were substantially effective only with ROCK-NM II inhibition. The PDX1+NKX6.1+ cells induced with NM II inhibitors were successfully engrafted and maturated in vivo. Taken together, these results suggest that NM IIs play inhibitory roles for the differentiation of hiPSCs to pancreatic endoderm cells. : Toyoda and colleagues show that ROCK or NM II inhibitors facilitate differentiation of PDX1+ posterior foregut cells into PDX1+NKX6.1+ pancreatic endoderm cells in hiPSC cultures. Consistently, NM II expression was elevated at low cell density, but upon aggregation, it was downregulated and associated with NKX6.1+ cell induction. The results provide mechanistic insight into the benefit of three-dimensional cultures for directed differentiation. Keywords: ESC, iPSC, differentiation, pancreas, pancreatic bud, pancreatic endoderm, NKX6.1, ROCK, non-muscle myosin II, cytoskeleton

Nitrogen (N) deposition is expected to influence forests. The effects of large-scale N fertilization on canopy layer insect–plant interactions in stands of tall, atmospheric nitrogen (N2)-fixing tree species have never been assessed. We conducted a large-scale fertilization experiment (100 kg N ha−1 year−1 applied to approximately 9 ha) over three years (2012–2014) in a cool temperate forest in northern Japan. Our goal was to evaluate relational responses between alder (Alnus hirsuta [Turcz.]) and their insect herbivores to N deposition. Specifically, we assessed leaf traits (N concentration, C:N ratio, condensed tannin concentration, and leaf mass per unit area (LMA)) and herbivory by three feeding guilds (leaf damage by chewers and the densities of gallers and miners) between the fertilized site and an unfertilized control. Fertilization led to increased galler density in spring 2013 and increased leaf damage by chewers in late summer 2014. For leaf traits, the LMA decreased in spring 2013 and late summer 2014, and the C:N ratio decreased in late summer 2013. The N and condensed tannin concentrations remained unchanged throughout the study period. There was a negative correlation between LMA and leaf damage by chewers, but LMA was not correlated with galler density. These results show that large-scale N fertilization had a positive plant-mediated (i.e., indirect) effect on leaf damage by chewers via a decrease in LMA in the canopy layer. Changes in physical defenses in canopy leaves may be a mechanism by which N fertilization affects the herbivory in tall N2-fixing trees.

In development, embryonic ectoderm differentiates into neuroectoderm and surface ectoderm using poorly understood mechanisms. Here, we show that the transcription factor OVOL2 maintains the transcriptional program of human corneal epithelium cells (CECs), a derivative of the surface ectoderm, and that OVOL2 may regulate the differential transcriptional programs of the two lineages. A functional screen identified OVOL2 as a repressor of mesenchymal genes to maintain CECs. Transduction of OVOL2 with several other transcription factors induced the transcriptional program of CECs in fibroblasts. Moreover, neuroectoderm derivatives were found to express mesenchymal genes, and OVOL2 alone could induce the transcriptional program of CECs in neural progenitors by repressing these genes while activating epithelial genes. Our data suggest that the difference between the transcriptional programs of some neuroectoderm- and surface ectoderm-derivative cells may be regulated in part by a reciprocally repressive mechanism between epithelial and mesenchymal genes, as seen in epithelial-to-mesenchymal transition.

Therapies are needed for the prevention of chondrocyte hypertrophy and thinning of articular cartilage, features of osteoarthritic joint destruction. Here, the authors show that interfering with Sik3 signalling can increase the size of the chondrocyte population and reduce severity of a surgically induced mouse model of osteoarthritis.

The aim of this study was to compare patient-reported symptoms of dry eye disease (DED) between the Japanese version of the Ocular Surface Disease Index (J-OSDI) and the Dry Eye-Related Quality-of-Life Score (DEQS). A total of 169 participants were enrolled between September 2017 and May 2018. Patients were administered the J-OSDI and DEQS questionnaires at their first (baseline) and follow-up visits to evaluate DED-related symptoms. The correlations between the J-OSDI total score and DEQS (Frequency and Degree) scores were evaluated using Pearson’s correlation coefficient, and their clinical differences were assessed using the Bland–Altman analysis. At the baseline visit, the J-OSDI score and DEQS (Frequency and Degree) were significantly correlated (r = 0.855, r = 0.897, respectively). Moreover, a significant correlation was found between the J-OSDI score and DEQS (Frequency and Degree) at the follow-up visit (r = 0.852, r = 0.888, respectively). The Bland–Altman analysis revealed a difference (bias) of 4.18 units at the baseline and 4.08 units at the follow-up between the scores of the two questionnaires. The J-OSDI and DEQS were significantly correlated with negligible score differences, suggesting that the J-OSDI can be reliably used for Japanese patients, allowing for cross-country comparisons.

Ultrasound measurements have been carried out to investigate the elastic and low-temperature 4f-electronic properties of the Sm-based 1-2-20 system SmPt2Cd20. It was found that SmPt2Cd20 shows a pronounced elastic softening toward the magnetic transition temperature Tc = 0.64 K in the temperature dependence of the transverse elastic constant CE = (C11 -C12)/2. The clear elastic softening behavior is suppressed gradually with increasing a magnetic field. The obtained results can be explained reasonably by a deformation potential approximation in which a coupling between conduction electrons and elastic strains caused by sound waves leads to a temperature dependence of elastic constants. This suggests possibly that the 4f electrons would become itinerant in the ground state in SmPt2Cd20. The nature of the low-temperature elastic and electronic states involving the ground state multiplets of Sm ions are discussed.

FePt alloy thin films with cap-layers of MgO or C are prepared on MgO(001) single-crystal substrates by using a two-step method consisting of low-temperature deposition at 200 °C followed by high-temperature annealing at 600 °C. The FePt film thickness is fixed at 10 nm, whereas the cap-layer thickness is varied from 1 to 10 nm. The influences of cap-layer material and cap-layer thickness on the variant structure and the L10 ordering are investigated. Single-crystal FePt(001) films with disordered fcc structure (A1) grow epitaxially on the substrates at 200 °C. Single-crystal MgO(001) cap-layers grow epitaxially on the FePt films, whereas the structure of C cap-layers is amorphous. The phase transformation from A1 to L10 occurs when the films are annealed at 600 °C. The FePt films with MgO cap-layers thicker than 2 nm consist of L10(001) variant with the c-axis perpendicular to the substrate surface, whereas those with C cap-layers involve small volumes of L10(100) and (010) variants with the c-axis lying in the film plane. The in-plane and the out-of-plane lattices are respectively more expanded and contracted in the continuous-lattice MgO/FePt/MgO structure due to accommodations of misfits of FePt film with respect to not only the MgO substrate but also the MgO cap-layer. The lattice deformation promotes phase transformation along the perpendicular direction and L10 ordering. The FePt films consisting of only L10(001) variant show strong perpendicular magnetic anisotropies and low in-plane coercivities. The present study shows that an introduction of epitaxial cap-layer is effective in controlling the c-axis perpendicular to the substrate surface.

Most patients complaining of pharyngeal pain have an upper respiratory tract infection or other local explanation for their pain. Here we show 3 rare cases of patients visiting our Otorhinolaryngology Department who had an initial symptom of pharyngeal pain caused by acute coronary syndrome (ACS). An electrocardiogram and a cardiac biomarker test are recommended to exclude ACS with atypical presentation in cases without pharyngolaryngeal findings comparable to pharyngeal pain.

Formation of L10-oredered structure from disordered A1 phase has been investigated for FePt and FePd films on MgO(001) substrates employing a two-step method consisting of low temperature deposition at 200 °C followed by high-temperature annealing at 600 °C. L10-(001) variant crystal with the c-axis perpendicular to the substrate grows preferentially in FePd films whereas L10-(100), (010) variants tend to be mixed with the L10-(001) variant in FePt films. The structure analysis by X-ray diffraction indicates that a difference in A1 lattice strain is the influential factor that determines the resulting L10-variant structure in ordered thin films. Misfit dislocations and anti-phase boundaries are observed in high-resolution transmission electron micrographs of 10 nm-thick Fe(Pt, Pd) film consisting of L10-(001) variants which are formed through atomic diffusion at 600 °C in a laterally strained FePt/PeFd epitaxial thin film. Based on the experimental results, a nucleation and growth model for explaining L10-variant formation is proposed, which suggests a possibility in tailoring the L10 variant structure in ordered magnetic thin films by controlling the alloy composition, the layer structure, and the substrate material.

A wearable Knee Assistive Instrument for the walk rehabilitation was newly developed. Especially, this system aimed at supporting the rehabilitation for the post-TKA (Total Knee Arthroplasty) which is a popular surgery for aging people. This system consisted of an assisting mechanism for the knee joint, a hip joint support system and a foot pressure sensor system. The driving system of this robot consisted of a CPU board which generated the walking pattern, a Li-ion battery, DC motors with motor drivers, contact sensors to detect the state of foot and potentiometers to detect the hip joint angle. The control method was proposed to reproduce complex motion of knee joint as much as possible, and to increase hip or knee flexion angle. Especially, this method used the timing that heel left from the floor. This method included that the lower limb was raised to prevent a subject's fall. Also, the prototype of knee assisting system was tested. It was confirmed that the assisting system is useful.

In this paper, we consider the following sliding puzzle called torus puzzle. In an m by n board, there are mn pieces numbered from 1 to mn. Initially, the pieces are placed in ascending order. Then they are scrambled by rotating the rows and columns without the player’s knowledge. The objective of the torus puzzle is to rearrange the pieces in ascending order by rotating the rows and columns. We provide a solution to this puzzle. In addition, we provide lower and upper bounds on the number of steps for solving the puzzle. Moreover, we consider a variant of the torus puzzle in which each piece is colored either black or white, and we present a hardness result for solving it.

Nature Communications 7: Article number: 10959 (2016); Published: 24 March 2016; Updated: 27 June 2016 In this Article, there is an error in the labelling of the western blot in Fig. 3f. The labels indicating the presence of pterosin B in each lane should read ‘−++ +’. The correct version of Fig. 3 appears below.

BACKGROUND: Recent studies have shown that in addition to brain (or B-type) natriuretic peptide (BNP) and the N-terminal proBNP fragment, levels of intact proBNP are also increased in heart failure. Moreover, present BNP immunoassays also measure proBNP, as the anti-BNP antibody cross-reacts with proBNP. It is important to know the exact levels of proBNP in heart failure, because elevation of the low-activity proBNP may be associated with the development of heart failure. METHODOLOGY/PRINCIPAL FINDINGS: We therefore established a two-step immunochemiluminescent assay for total BNP (BNP+proBNP) and proBNP using monoclonal antibodies and glycosylated proBNP as a standard. The assay enables measurement of plasma total BNP and proBNP within only 7 h, without prior extraction of the plasma. The detection limit was 0.4 pmol/L for a 50-µl plasma sample. Within-run CVs ranged from 5.2%-8.0% in proBNP assay and from 7.0%-8.4% in total BNP assay, and between-run CVs ranged from 5.3-7.4% in proBNP assay and from 2.9%-9.5% in total BNP assay, respectively. The dilution curves for plasma samples showed good linearity (correlation coefficients = 0.998-1.00), and analytical recovery was 90-101%. The mean total BNP and proBNP in plasma from 116 healthy subjects were 1.4 ± 1.2 pM and 1.0 ± 0.7 pM, respectively, and were 80 ± 129 pM and 42 ± 70 pM in 32 heart failure patients. Plasma proBNP levels significantly correlate with age in normal subjects. CONCLUSIONS/SIGNIFICANCE: Our immunochemiluminescent assay is sufficiently rapid and precise for routine determination of total BNP and proBNP in human plasma.

BackgroundFor the diagnosis of seasonal influenza, clinicians rely on point-of-care testing (POCT) using commercially available kits developed against seasonal influenza viruses. However, POCT has not yet been established for the diagnosis of pandemic influenza A virus (H1N1pdm) infection due to the low sensitivity of the existing kits for H1N1pdm.Methodology/principal findingsAn immunochromatography (IC) test kit was developed based on a monoclonal antibody against H1N1pdm, which does not cross-react with seasonal influenza A or B viruses. The efficacy of this kit (PDM-IC kit) for the diagnosis of H1N1pdm infection was compared with that of an existing kit for the detection of seasonal influenza viruses (SEA-IC kit). Nasal swabs (n = 542) were obtained from patients with flu-like syndrome at 13 clinics in Osaka, Japan during the winter of 2010/2011. Among the 542 samples, randomly selected 332 were further evaluated for viral presence by reverse transcriptase polymerase chain reaction (RT-PCR). The PDM-IC kit versus the SEA-IC kit showed higher sensitivity to and specificity for H1N1pdm, despite several inconsistencies between the two kits or between the kits and RT-PCR. Consequently, greater numbers of false-negative and false-positive cases were documented when the SEA-IC kit was employed. Significant correlation coefficients for sensitivity, specificity, and negative prediction values between the two kits were observed at individual clinics, indicating that the results could be affected by clinic-related techniques for sampling and kit handling. Importantly, many patients (especially influenza-negative cases) were prescribed anti-influenza drugs that were incongruous with their condition, largely due to physician preference for patient responses to questionnaires and patient symptomology, as opposed to actual viral presence.Conclusions/significanceConcomitant use of SEA-IC and PDM-IC kits increased the likelihood of correct influenza diagnosis. Increasing the credibility of POCT is anticipated to decrease the inappropriate dispensing of anti-influenza drugs, thereby minimizing the emergence of drug-resistant H1N1pdm strains.

Identification of regulatory elements within the genome is crucial for understanding the mechanisms that govern cell type-specific gene expression. We generated genome-wide maps of open chromatin sites in 3T3-L1 adipocytes (on day 0 and day 8 of differentiation) and NIH-3T3 fibroblasts using formaldehyde-assisted isolation of regulatory elements coupled with high-throughput sequencing (FAIRE-seq). FAIRE peaks at the promoter were associated with active transcription and histone modifications of H3K4me3 and H3K27ac. Non-promoter FAIRE peaks were characterized by H3K4me1+/me3-, the signature of enhancers, and were largely located in distal regions. The non-promoter FAIRE peaks showed dynamic change during differentiation, while the promoter FAIRE peaks were relatively constant. Functionally, the adipocyte- and preadipocyte-specific non-promoter FAIRE peaks were, respectively, associated with genes up-regulated and down-regulated by differentiation. Genes highly up-regulated during differentiation were associated with multiple clustered adipocyte-specific FAIRE peaks. Among the adipocyte-specific FAIRE peaks, 45.3% and 11.7% overlapped binding sites for, respectively, PPARγ and C/EBPα, the master regulators of adipocyte differentiation. Computational motif analyses of the adipocyte-specific FAIRE peaks revealed enrichment of a binding motif for nuclear family I (NFI) transcription factors. Indeed, ChIP assay showed that NFI occupy the adipocyte-specific FAIRE peaks and/or the PPARγ binding sites near PPARγ, C/EBPα, and aP2 genes. Overexpression of NFIA in 3T3-L1 cells resulted in robust induction of these genes and lipid droplet formation without differentiation stimulus. Overexpression of dominant-negative NFIA or siRNA-mediated knockdown of NFIA or NFIB significantly suppressed both induction of genes and lipid accumulation during differentiation, suggesting a physiological function of these factors in the adipogenic program. Together, our study demonstrates the utility of FAIRE-seq in providing a global view of cell type-specific regulatory elements in the genome and in identifying transcriptional regulators of adipocyte differentiation.