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Pancreatic ductal adenocarcinoma (PDAC) is characterized by a tumor immune microenvironment (TIME) that promotes resistance to immunotherapy. A preclinical model system that facilitates studies of the TIME and its impact on the responsiveness of human PDAC to immunotherapies remains an unmet need. We report a novel mouse model, which develops metastatic human PDAC that becomes infiltrated by human immune cells recapitulating the TIME of human PDAC. The model may serve as a versatile platform to study the nature of human PDAC TIME and its response to various treatments.

The fear and devastation caused by the COVID-19 pandemic has been mitigated by the successful development and deployment of prophylactic vaccines that substantially lowered the incidences of symptomatic infection, hospitalization, and death. However, as the causative agent SARS-CoV-2 continues to spread and evolve worldwide, vaccine-breakthrough infections have become frequent, especially after the emergence of viral variants that are antigenically distant from the ancestral strain used in the current vaccines. Additional approaches are therefore needed in our prevention tool kit. Here, we report on a glycolipid termed 7DW8-5 that exploits the host innate immune system to enable rapid control of viral infections in vivo. This glycolipid binds to CD1d on antigen-presenting cells and thereby stimulates NKT cells to release a cascade of cytokines and chemokines. The intranasal administration of 7DW8-5 prior to virus exposure significantly blocked infection by three different authentic variants of SARS-CoV-2, as well as by respiratory syncytial virus and influenza virus, in mice or hamsters. We also found that this protective antiviral effect is both host-directed and mechanism-specific, requiring both the CD1d molecule and interferon- gamma. A chemical compound like 7DW8-5 that is easy to administer and cheap to manufacture may be useful not only in slowing the spread of COVID-19 but also in responding to future pandemics that include currently ongoing influenza, SARS-CoV-2 and respiratory syncytia virus- associated tripledemic long before vaccines or drugs are developed.

BACKGROUNDObsessive-compulsive disorder (OCD) is a psychiatric disorder characterized by obsessive thoughts and compulsive behavior. While some theories imply that OCD patients have cognitive biases and dysfunctional motivation regarding a potential threat, these views are not adequately supported by neurological evidence. Hypothalamic perifornical (PeF) urocortin-3 (UCN3) neurons are involved in defensive responses to a potential threat, and the activation of these neurons in mice induces repetitive and excessive checking and burying of novel objects. In this study, we evaluated the hypothesis that mice in which PeF UCN3 neurons are activated can serve as an OCD model.METHODSPeF UCN3 neurons were chemogenetically activated with clozapine-N oxide (CNO) inUcn3-Cremice. Marble-burying activity, repetitive/stereotypic behaviors in the homecage, and excessive responses to a novel object were measured as OCD-like behaviors. The effects of clinically used drugs for treating OCD on these behaviors were evaluated. The effect of CNO on neural activity in the cortico-striato-thalamo-cortical loop (which is regarded as an OCD circuit) was assessed with c-Fos immunolabeling.RESULTSCNO increased marble-burying activity, evoked homecage-specific repetitive/stereotypic behaviors that probably aimed to seal entrances, and induced repetitive and excessive checking and burying of novel objects. These behaviors were suppressed by selective serotonin-reuptake inhibitors but not by diazepam. CNO increased neural activity in the cortico-striato-thalamo-cortical loop.CONCLUSIONSThese results indicated that mice whose PeF UCN3 neurons are activated can serve as a model of OCD, particularly as a checking model. This supports theories concerning the role of potential threats in the pathophysiology of OCD.

The fear and devastation caused by the COVID-19 pandemic has been mitigated by the successful development and deployment of prophylactic vaccines that substantially lowered the incidences of symptomatic infection, hospitalization, and death. However, as the causative agent SARS-CoV-2 continues to spread and evolve worldwide, vaccine-breakthrough infections have become frequent, especially after the emergence of viral variants that are antigenically distant from the ancestral strain used in the current vaccines. Additional approaches are therefore needed in our prevention tool kit. Here, we report on a glycolipid termed 7DW8-5 that exploits the host innate immune system to enable rapid control of viral infections in vivo. This glycolipid binds to CD1d on antigen-presenting cells and thereby stimulates NKT cells to release a cascade of cytokines and chemokines. The intranasal administration of 7DW8-5 prior to virus exposure significantly blocked infection by three different variants of SARS-CoV-2, as well as by respiratory syncytial virus and influenza virus, in mice or hamsters. We also found that this protective antiviral effect is both host-directed and mechanism-specific, requiring both the CD1d molecule and interferon-γ. A chemical compound like 7DW8-5 that is easy to administer and cheap to manufacture may be useful not only in slowing the spread of COVID-19 but also in responding to future pandemics due to a respiratory virus long before vaccines or drugs are developed.

International labor mobility is a key factor for a well-functioning labor market. Although educational attainment is known to affect regional labor mobility within a country, evidence of a relationship between schooling and international labor mobility is limited, particularly in developing countries. This study uses the across-cohort variation in the exposure to the 1988 free secondary education reform in the Philippines to examine the impact of years of education on the propensity of working abroad. The results suggest that free secondary education increased the years of education for men. Moreover, the additional years of education reduced the likelihood of working abroad by 3.2% points on average. However, an extra year of female education was not associated with the probability of working abroad. These results indicate that a program for improving access to secondary education may affect international labor mobility for men even after a few decades. It underscores the importance of considering the possible labor market consequences when designing the education reform in developing countries.

Significance Obesity is now considered an epidemic problem worldwide. To develop effective interventions against obesity, we need to understand precisely the regulatory mechanism of adipogenesis, a process of forming adipocytes from fibroblast cells. In this study, we revealed that the Arid5a protein regulates the adipogenesis and the development of obesity in mice. Arid5a inhibits adipogenesis by suppressing the transcription of peroxisome proliferator activated receptor gamma 2 (Ppar-γ2), which is the main facilitator of adipogenesis. Further, we showed adipose tissue homeostasis is maintained by the coordinated activity of Arid5a and Ppar-γ2. Our study provides important insights into the novel mechanism of adipose tissue homeostasis., Immune cells infiltrate adipose tissues and provide a framework to regulate energy homeostasis. However, the precise underlying mechanisms and signaling by which the immune system regulates energy homeostasis in metabolic tissues remain poorly understood. Here, we show that the AT-rich interactive domain 5A (Arid5a), a cytokine-induced nucleic acid binding protein, is important for the maintenance of adipose tissue homeostasis. Long-term deficiency of Arid5a in mice results in adult-onset severe obesity. In contrast, transgenic mice overexpressing Arid5a are highly resistant to high-fat diet-induced obesity. Inhibition of Arid5a facilitates the in vitro differentiation of 3T3-L1 cells and fibroblasts to adipocytes, whereas its induction substantially inhibits their differentiation. Molecular studies reveal that Arid5a represses the transcription of peroxisome proliferator activated receptor gamma 2 (Ppar-γ2) due to which, in the absence of Arid5a, Ppar-γ2 is persistently expressed in fibroblasts. This phenomenon is accompanied by enhanced fatty acid uptake in Arid5a-deficient cells, which shifts metabolic homeostasis toward prolipid metabolism. Furthermore, we show that Arid5a and Ppar-γ2 are dynamically counterregulated by each other, hence maintaining adipogenic homeostasis. Thus, we show that Arid5a is an important negative regulator of energy metabolism and can be a potential target for metabolic disorders.

Although degree of T-cell infiltration in CRC was shown to correlate with a positive prognosis, the contribution of phenotypically and functionally distinct T cell subtypes within tumors remains unclear. We analyzed 37,931 single T cells with respect to transcriptome, TCR sequence and 23 cell surface proteins, from tumors and adjacent normal colon of 16 patients. Our comprehensive analysis revealed two phenotypically distinct cytotoxic T cell populations within tumors, including positively prognostic effector memory cells and non-prognostic resident memory cells. These cytotoxic T cell infiltrates transitioned from effector memory to resident memory in a stage-dependent manner. We further defined several Treg subpopulations within tumors. While Tregs overall were associated with positive clinical outcomes, CD38+ peripherally-derived Tregs, phenotypically related to Th17 cells, correlated with poor outcomes independent of cancer stage. Thus, our data highlight the diversity of T cells in CRC and demonstrate the prognostic significance of distinct T cell subtypes, which could inform therapeutic strategies.

This paper examines the impact of female education on adolescent fertility, the health status of their children, and the mechanism through which education affects these outcomes. To address the endogeneity of educational attainment, we utilise Universal Primary Education policy (UPE) in Uganda. Education is instrumented by the interaction between across-cohorts differences in exposure to UPE and the differences in its effective benefits across districts with varying pre-programme rates of completing primary education. We particularly focus on the fully treated cohorts whose fees were abolished before they entered school. Results show that attending an additional year of schooling reduces the probability of marriage and that of giving birth before age 18 by 7.0–7.2 percentage points. Among those who become mothers, educated women use maternal care and infant immunisation more often, and had lower probability that their child dies before 18 months after birth. These effects are likely to arise because educated women tend to be literate and prefer to have fewer children. They exhibit better knowledge about reproductive issues. Weak evidence is found for an increase in the probability of working in the non-agricultural sector. No evidence is, however, found for assortative mating, and evidence for improved bargaining power is mixed.

Recently, studies of intestinal microbiota have been conducted using mainly next-generation sequencers to perform comprehensive bacterial DNA analyses. When using this molecular biological approach, intestinal bacterial DNA is extracted from fecal samples. But the influence of the fecal sample storage condition and the methods of DNA extraction on the analysis have not been investigated as far as we know. In this study, we evaluate the effects of different freezing conditions and storage periods of microbial DNA in fecal samples using PCR-DGGE analysis. Fecal samples were stored at −20 ºC, −80 ºC and −80 ºC followed by a liquid nitrogen treatment and kept for 3 months and 1 year, respectively. Microbial DNA extracted from these fecal samples was examined using PCR-DGGE analysis to monitor total intestinal microbiota: Bacteroides, Bifidobacterium, Lactobacillus and Clostridium groups. DGGE profiles demonstrated that the total bacterial flora was stable and no significant changes were found due to storage conditions or periods. In genus specific detection of samples stored for three months, DNA bands were detected in all samples except for in part of the Clostridium group. In the case of fecal samples stored for one year, both at −80 ºC and also treated with liquid nitrogen, amplified genus specific bands were present in all samples. A different band pattern was observed only in the amplicon of the liquid nitrogen treated samples from the Clostridium group. On the other hand, in microbial DNA extracted from samples preserved at −20 ºC it was impossible to amplify specific fragments. Since some bacterial groups in fecal samples were affected by the freezing method, storage conditions and period, it appears that rapidly freezing fecal samples may be the most effective way to maintain intestinal microbiota.

AT-rich interactive domain-containing protein 5a (Arid5a) is an RNA-binding protein (RBP) required for autoimmunity via stabilization of interleukin-6 (Il6) and signal transducer and activator of transcription 3 (STAT3) mRNAs. However, the roles of Arid5a in Th17 cells and its association with autoimmunity remain unknown. Here, we show that the levels of Arid5a and OX40 are correlated in CD4+ T cells under Th17 conditions in an IL-6-dependent manner. Lack of Arid5a in T cells reduced OX40 expression levels and repressed IL-17 production in response to OX40 ligation. Arid5a stabilized OX40 mRNA by recognizing the alternative decay element (ADE)-like stem-loop (SL) in the 3' untranslated region (3'UTR). Interestingly, Arid5a impaired the RNA-destabilizing functions of Regnase-1 and Roquin-1 on OX40 ADE-like SL. In EAE, Arid5a-deficient mice exhibited resistance to EAE, with reduced OX40 expression in CD4+ T cells, and the number of CD4+ CD45+ T cells was decreased in CNS. Furthermore, ameliorated EAE was induced by adoptive transfer of Arid5a-/- encephalitogenic CD4+ T cells expressing less OX40 mRNA and producing less IL-17. In conclusion, our findings indicate that the Arid5a/OX40 axis in CD4+ T cells may have important implications in pathogenesis of autoimmune diseases such as EAE.

The AT-rich interactive domain-containing protein 5a (Arid5a) plays a critical role in autoimmunity by regulating the half-life of Interleukin-6 (IL-6) mRNA. However, the signaling pathways underlying Arid5a-mediated regulation of IL-6 mRNA stability are largely uncharacterized. Here, we found that during the early phase of lipopolysaccharide (LPS) stimulation, NF-κB and an NF-κB-triggered IL-6-positive feedback loop activate Arid5a gene expression, increasing IL-6 expression via stabilization of the IL-6 mRNA. Subsequently, mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) promotes translocation of AU-rich element RNA-binding protein 1 (AUF-1) from the nucleus to the cytoplasm, where it destabilizes Arid5a mRNA by binding to AU-rich elements in the 3΄ UTR. This results in downregulation of IL-6 mRNA expression. During the late phase of LPS stimulation, p38 MAPK phosphorylates Arid5a and recruits the WW domain containing E3 ubiquitin protein ligase 1 (WWP1) to its complex, which in turn ubiquitinates Arid5a in a K48-linked manner, leading to its degradation. Inhibition of Arid5a phosphorylation and degradation increases production of IL-6 mRNA. Thus, our data demonstrate that LPS-induced NF-κB and MAPK signaling are required to control the regulation of the IL-6 mRNA stabilizing molecule Arid5a. This study therefore substantially increases our understanding of the mechanisms by which IL-6 is regulated.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are among the major causes of death worldwide due to acute inflammation in the lung. AT-rich interactive domain–containing 5a (Arid5a) is an RNA-binding protein involved in inflammatory autoimmune disease through post-transcriptional control of Il6, Stat3 and Tbx21 gene expression. We found that Arid5a-deficient mice were highly refractory to bleomycin (BLM)-induced lethality. Arid5a deficiency suppressed lung pathology, cytokine production (especially, IL-6), and clinical symptoms in BLM-treated mice. Production of reactive oxygen species (ROS) in response to BLM-induced cellular damage was inhibited in Arid5a-deficient mice, potentially affecting the level of oxidized 1-palmitoyl-2-arachidonoyl-phosphaticylcholine (OxPAPC) production. OxPAPC, which is supposed to be a TLR4/TLR2 ligand, stimulated expression of the Arid5a and Il6 genes. Thus, reduction of ROS production in Arid5a-deficient mice could mitigate OxPAPC production, which in turn decreases IL-6 production in vivo due to dysregulated post-transcriptional regulation by loss of Arid5a. Therefore, the control of Arid5a expression represents a potential therapeutic target for treatment of ALI and ARDS.

The association between schooling and religious beliefs has been widely documented in the social science literature. Evidence of a causal relationship is, however, limited, particularly in developing countries where religion still plays a significant role in politics and legislation. To bridge this gap in the literature, the present study uses the across cohort variations in the exposure to 1978 education reform in Indonesia to examine the impact of completed years of education on individual religiosity in later life. The results suggest that attaining another year of schooling reduces self-reported religiousness by four percentage points. Consistent with this finding, it also changes solitary religious acts by reducing the number of times individuals pray and the likelihood that they eat only halal food. Educational attainment, however, has little effect on participation in social religious activities or attitude toward other faiths, although it does reduce the religious influence on voting behavior. These results suggest that a program promoting educational attainment in Islamic countries may have an important impact on individual’s religiosity and country’s political economy in the long run.

Early childhood development relies on various micronutrients. We recently reported that home fortification of complementary foods using spirulina reduced the time to attain motor milestones in Zambian infants. The objective of this study is to estimate the long-term associations between spirulina supplementation during the first 1000 days and child gross motor development, fine motor development, language, and personal–social skills at preschool age. We used longitudinal data from a randomized trial conducted in Zambia. In 2015, 501 infants (age, 6–18 months) were provided daily supplements of maize-soy-based porridge with spirulina (SP) and without spirulina (CON). Supplementation period lasted for 16 months. In January 2018, children who participated in the initial trial were resurveyed (CON: 182 children; SP: 188 children; now aged 36–48 months). We assessed the infants’ gross motor development, fine motor development, language, and personal–social skills using a modified version of Malawi Development Assessment Tool. The initial clinical trial registration number was NCT03523182. Children in the SP group had higher scores in gross and fine motor development, language, and social skills than those in the CON group. Home fortification of complementary foods using spirulina during the first 1000 days improved development among Zambian children at preschool age.

Background: An estimated 216 million cases of malaria occurred worldwide every year. Cross sectional studies have reported negative association between maternal education and child malaria risks, but no randomized trial has confirmed a causal relationship between these two factors. I utilized the free primary education reform in Uganda to assess the causal effects of maternal schooling on the child's risk of malaria infection. Methods: Malaria biomarkers of children aged < five years were collected from the 2009 and 2014 Uganda Malaria Indicator Surveys (N = 5,316). In 1997, the government eliminated tuition requirement in primary schools and this increased the educational attainment of the affected cohorts. Using exposure to the reform as an instrumental variable, I used a two-stage least squares approach to estimate the causal effects of maternal year of education on the probability that a child would contract malaria at the time of the survey. I also evaluated the cost-effectiveness of primary schooling as a malaria control intervention. Findings: One extra year of maternal education reduced child's risk of malaria infection by 7·5 percentage points (p=0·057), from a baseline 34·6% among the children of 1980 pre-reform cohort. The length of maternal education was also positively associated with insecticide-treated bednets usage. The results were robust to a variety of sensitivity tests. Primary schooling for women was a cost-effective intervention to reduce child's malaria infection. Interpretation: Improving access to primary education could be a cost-effective measure to reduce malaria prevalence among children aged < five years in malaria-endemic countries. Funding: None. Declaration of Interest: None exist. Ethical Approval: The present study was reviewed by the Hitotsubashi University Institutional Review Board and considered exempt from full ethics review because the analysis was based on a publicly available, anonymized dataset.

To investigate the prebiotic effect of lactulose at low dosages, we assessed changes in defaecation frequency following ingestion of 1, 2, or 3 g/day of lactulose for 2 weeks. Each test was carried out after a 2-week washout period. This was an open-label, before-after trial that enrolled 26 healthy Japanese women. The defaecation frequency, number of defaecation days, and number of faecal bifidobacteria increased significantly compared with before ingestion of 1, 2, and 3 g/day of lactulose. These results suggest that even 1 g/day of lactulose could have a prebiotic effect.

Masuda et al. show that Arid5a regulates the fate of naive CD4+ T cells to pro- or antiinflammatory T cells through selective stabilization of Stat3 mRNA under Th17-polarizing conditions., Balance in signal transducer and activator of transcription (STAT) activation is a key factor in regulating the fate of naive CD4+ T cells. Here, we demonstrate that AT-rich interactive domain-containing protein 5a (Arid5a) in T cells directs naive CD4+ T cells to differentiate into inflammatory CD4+ T cells, especially Th17 cells, through selective stabilization of Stat3 (but not Stat1 and Stat5) mRNA in an IL-6–dependent manner. Loss of Arid5a in T cells led to reduction of STAT3 level under Th17-polarizing conditions, whereas STAT1 and STAT5 in Arid5a-deficient T cells were highly activated compared with those of WT T cells under the same conditions. These cells displayed the feature of antiinflammatory (Il10-expressing) CD4+ T cells. Thus, we show a T cell–intrinsic role of Arid5a on fate decisions of naive CD4+ T cells through selective stabilization of Stat3 mRNA.

Following a large outbreak of Shiga toxin-producing Escherichia coli (STEC) O157 infection in 2012, the prerequisite program for the production of pickles in Japan was revised to include disinfection with sodium hypochlorite (NaClO). Here, we examined the indicator bacterial counts, incidence of STEC and Salmonella spp. (by polymerase chain reaction [PCR] screening), and bacterial community composition (by 16S rRNA pyrosequencing analysis) from the intermediate products and the related facilities at a collaborative factory to evaluate the dynamics of microbial quality in lightly pickled vegetables during the manufacturing process following the revised program. Plate counts showed a significant reduction in coliform counts throughout processing, whereas the reduction in total viable counts was relatively less than that of coliforms. No STEC and Salmonella spp. were recovered from any of the samples. 16S rRNA pyrosequencing analysis revealed the process-by-process alteration of bacterial community composition in which the yields of Pseudomonas spp. were drastically affected by soaking in high concentration of NaCl. In summary, we demonstrate that the revised prerequisite program is indeed functional to reduce the microbial risks. Practical Applications Fresh or minimally processed produce are common vehicles of foodborne pathogens that have been responsible for several outbreaks worldwide. Through the use of indicator bacterial counts in combination with a metagenomic approach, we could trace the process-by-process dynamics of bacterial community composition. Information and relief provided to manufacturers/consumers could help improve public health and food safety.

The participation of a specific subset of B cells and how they are regulated in cancer is unclear. Here, we demonstrate that the proportion of CD5+ relative to interleukin-6 receptor α (IL-6Rα) expressing B cells was greatly increased in tumors. CD5+ B cells responded to IL-6 in the absence of IL-6Rα. IL-6 directly bound to CD5, leading to activation of the transcription factor STAT3 via gp130 and its downstream kinase JAK2. STAT3 upregulated CD5 expression, thereby forming a feed-forward loop in the B cells. In mouse tumor models, CD5+ but not CD5−B cells promoted tumor growth. CD5+ B cells also showed activation of STAT3 in multiple types of human tumor tissues. Thus, our findings demonstrate a critical role of CD5+ B cells in promoting cancer.

Background: Infection, tissue damage and aging can cause inflammation with high levels of inflammatory cytokines. Overproduction of inflammatory cytokines often leads to systemic inflammatory response syndrome (SIRS), severe sepsis, and septic shock. However, prominent therapeutic targets have not been found, although the incidence of sepsis is likely to increase annually. Our recent studies indicate that some RNA-binding proteins, which control gene expression of inflammatory cytokines at the post-transcriptional level, may play a critical role in inflammatory diseases such as sepsis. Results: 1) One of the RNA-binding proteins, AT-rich interactive domain-containing 5a (Arid5a) promotes cytokine production through control of mRNA half-lives of pro-inflammatory molecules such as IL-6, STAT3, T-bet, and OX40 in activated macrophages and T cells. Arid5a KO mice are refractory to endotoxin shock, bleomycininduced lung injury, and inflammatory autoimmune disease. 2) Chlorpromazine (CPZ), which is recognized as a psychotic drug, impairs post-transcriptional gene expression of Il6 in LPS-stimulated macrophages: CPZ inhibits the binding activity of Arid5a to the 3’UTR of Il6 mRNA, thereby destabilizing Il6 mRNA possibly through suppression of Arid5a expression. 3) CPZ has strong suppressive effects on cytokine production such as TNF-α in vivo. Mice with treatment of CPZ are resistant to lipopolysaccharide (LPS)-induced shock. Conclusion: Thus, Arid5a contributes to the activation of macrophages and T cells through positive control of mRNA half-lives of inflammatory cytokines and its related molecules, which might lead to cytokine storm. Interestingly, Arid5a was identified from an inhibitory effect of CPZ on IL-6 production in macrophages activated by LPS. Therefore, CPZ derivatives or Arid5a inhibitors may have a potential to suppress severe sepsis through control of post-transcriptional gene expression.

Significance Immune cell activation is accompanied by dynamic changes in expression of genes related to inflammation. Concomitantly, immune reactions are tightly controlled to prevent harmful pathologies due to sustained inflammation. Gene expression is controlled at multiple checkpoints. Among these, the post-transcriptional regulation of the balance between Arid5a and Regnase-1 is important for the induction of inflammation. Our findings provide important insight into the role of the regulation of nucleocytoplasmic localization of Arid5a in the development of inflammation through the induction of a change in the ratio of Arid5a to Regnase-1., Adenine-thymine (AT)-rich interactive domain 5a (Arid5a) is an RNA-binding protein found in the cytoplasm and nucleus of normally growing cells. Although Arid5a is known to play an important role in immune regulation, whether and how Arid5a subcellular localization impacts immune regulation has remained unclear. In this study, we generated Arid5a transgenic (TG) mice to address this question. While ectopic Arid5a overexpression did not affect expression of inflammatory cytokines under unstimulated conditions, significantly higher levels of inflammatory cytokines, such as IL-6, were produced in response to lipopolysaccharide (LPS) stimulation. Consistent with this, TG mice were more sensitive to LPS treatment than wild-type mice. We also found that Arid5a is imported into the nucleus via a classical importin-α/β1–mediated pathway. On stimulation, nuclear Arid5a levels were decreased, while there was a concomitant increase in cytoplasmic Arid5a. Arid5a is associated with up-frameshift protein 1, and its nuclear export is regulated by a nuclear export receptor, chromosomal region maintenance 1. Taken together, these data indicate that Arid5a is a dynamic protein that translocates to the cytoplasm from the nucleus so as to properly exert its dual function in mRNA stabilization and transcriptional regulation during inflammatory conditions.

AT-rich interactive domain-containing protein 5a (Arid5a) is an RNA-binding protein (RBP) required for autoimmunity via stabilization of interleukin-6 (Il6) and signal transducer and activator of transcription 3 (STAT3) mRNAs. However, the roles of Arid5a in Th17 cells and its association with autoimmunity remain unknown. Here, we show that the levels of Arid5a and OX40 are correlated in CD4

Checkpoint blockade immunotherapy has shown great success in certain types of cancers, but are ineffective in most cancers. We do not understand why these therapies fail. Here, we seek to better understand functional changes in T cells in responsive tumors through single cell TCR sequencing and high dimensional phenotypic analysis within single tumor-infiltrating T cells. Through surgical biopsy, we analyze the same tumor longitudinally before and after treatment, enabling us to track the fate of individual T cell clones. Our data show that the CD4/CD8 ratio decreases upon treatment. T cells expressing inhibitory markers such as PD1, CTLA4, diminish post treatment. We see distinct clusters of effector CD4 and CD8 T cells in treated tumors, including effector CD4 populations with Tfh, Th1 and Th2 phenotypes. CD4 Tregs are greatly diminished upon successful treatment, and diminish through clonal contraction. Furthermore, TCR acts as a unique marker for T cell ancestry, which links T cell function to T cell specificity. Based on TCR clonality, we observe that expanded T cell clones before treatment do not share the same TCRs with expanded T cell clones after treatment. This implies new CD8 T cell clones are recruited into the tumor upon treatment. From our data, we will be able to identify T cell clones that are phenotypically distinct and expanded in responsive tumors which is critical in understanding what makes for a successful T cell response after immunotherapy.

For the surveillance of the prevalence of Campylobacter jejuni and Campylobacter coli in raw chicken products in Japan, a qualitative method, National Institute of Health Sciences Japan (NIHSJ)-02, was developed as an alternative to International Organization for Standardization (ISO) 10272-1:2006. In the NIHSJ-02 culture method, the enrichment step is carried out in a reduced volume of Preston broth at 42 ± 1°C to reduce cost and space, and to prevent the overgrowth of background bacteria. To evaluate the performance of NIHSJ-02, a collaborative study was conducted, and the results obtained by NIHSJ-02 were compared with those obtained using the reference method, ISO 10272-1:2006. Fifteen laboratories participated; each examined 48 minced chicken samples consisting of test samples uninoculated, inoculated with C. jejuni at a low or high level, and inoculated with C. coli at a low level. The average probabilities of detection by NIHSJ-02 across laboratories were 0.033, 0.222, 0.678, and 0.267 in samples uninoculated, inoculated with C. jejuni at a low and high level, and with C. coli at a low level, respectively. Those by ISO 10272-1:2006 were 0.051, 0.128, 0.551, and 0.090. Significantly higher probabilities of detection were determined by NIHSJ-02 compared to ISO 10272-1:2006, except for uninoculated samples. On the other hand, significantly lower frequency of occurrence of background bacteria was observed by NIHSJ-02 (43.1%) compared with ISO 10272-1:2006 (92.6%). NIHSJ-02 showed better performance than ISO 10272-1:2006 with regard to the selective detection of C. jejuni and C. coli in chicken.

Interleukin-6 (IL-6), originally identified as a B cell stimulatory factor-2, is a typical cytokine featuring redundancy and pleiotropic activity. A transient expression of IL-6 participates in host defense against acute environmental stress such as infections and injuries by activating immune responses, hematopoiesis and acute phase reactions. However, its abnormal, persistent production plays an important pathological role in the development of various autoimmune inflammatory diseases, so that it was hypothesized that IL-6 blockade would constitute a novel strategy for the treatment of such diseases and to this purpose tocilizumab, a humanized antiIL-6 receptor monoclonal antibody, was developed. Clinical trials have indeed proved the efficacy and tolerable safety of tocilizumab for patients with moderate to severe rheumatoid arthritis, and it is now used as a“ “ “ “made-in-Japan”innovative biologic for rheumatoid arthritis in more than 90 countries worldwide, as well as for patients with Castleman's disease and systemic and polyarticular juvenile idiopathic arthritis. Moreover, favorable results of recent clinical trials or case reports of off-label use with tocilizumab indicate that it is likely to be broadly applicable for the treatment of various autoimmune inflammatory diseases. Its wider application for various diseases as well as clarification of the mechanism(s) through which IL-6 blockade becomes clinically efficacious and of the etiology of dysregulated persistent IL-6 production in such diseases are important issues for future studies.

Interleukin-6 (IL-6) is a prototypical cytokine with functional pleiotropy and plays an important role in host defense. When infections or tissue injuries occur, IL-6 is promptly produced by monocytes and macrophages and contributes to removal of infectious agents and restoration of damaged tissues through activation of immune, hematological, and acute-phase responses. Once stress is removed from the host, IL-6 synthesis ends, but uncontrolled excessive or persistent IL-6 production plays a pathological role in the development of various inflammatory diseases and cancers, indicating that IL-6 is a double-edged sword for the host. Thus, the proper IL-6 expression is very important for host defense and is strictly controlled by chromatin structure, transcriptional regulation, and posttranscriptional modification. Differentiation status of cells, various transcription factors, RNA-binding proteins, and microRNAs are involved in this process. Since it is assumed that dysregulation of any of these regulatory molecules may cause abnormal IL-6 expression in a particular disease, further elucidation of the factors and processes involved in IL-6 expression can be expected to facilitate to clarification of pathogenesis and to identification of novel target molecule(s) for specific diseases.

Adenine-thymine (AT)-rich interactive domain containing protein 5a (Arid5a) is an RNA-binding protein that has been shown to play an important immune regulatory function via the stabilization of IL-6 and STAT3 mRNA. However, the role of Arid5a in the overwhelming and uncontrolled immune response that leads to septic shock is unknown. Here, we report that Arid5a-deficient mice are highly resistant to lipopolysaccharide (LPS)-induced endotoxic shock and secrete lower levels of major proinflammatory cytokines, including IFN-γ, IL-6, and TNF-α, than WT mice in response to LPS. Arid5a deficiency resulted in decreased levels of IFN-γ under Th1 cell conditions, in which T-box expressed in T cells (T-bet) mRNA expression was inhibited. Arid5a bound to the conserved stem loop structure of the 3′UTR of T-bet and stabilized its mRNA. Arid5a-deficient mice were also resistant to Propionibacterium acnes-primed LPS injection, which is considered to be a T-cell–mediated IFN-γ dependent endotoxic shock mouse model. Thus, regulation of IFN-γ by Arid5a via the stabilization of T-bet mRNA in Th1 cells contributes to the development of septic shock in mice. In addition, our previous study suggests that Arid5a control the IL-6 level in vivo in response to LPS by stabilization of IL-6 mRNA. We also observed that neutralization of IFN-γ and IL-6 significantly recovered the mice from endotoxic shock. Taken together, we conclude that Arid5a regulates the augmentation of IL-6 and IFN-γ in response to LPS, which possibly works synergistically for amplification of various other cytokines that ultimately cause the development of septic shock in mice.

Early childhood development relies on various micronutrients. We recently reported that home fortification of complementary foods using spirulina reduced the time to attain motor milestones in Zambian infants. The objective of this study is to estimate the long-term associations between spirulina supplementation during the first 1000 days and child gross motor development, fine motor development, language, and personal&ndash, social skills at preschool age. We used longitudinal data from a randomized trial conducted in Zambia. In 2015, 501 infants (age, 6&ndash, 18 months) were provided daily supplements of maize-soy-based porridge with spirulina (SP) and without spirulina (CON). Supplementation period lasted for 16 months. In January 2018, children who participated in the initial trial were resurveyed (CON: 182 children, SP: 188 children, now aged 36&ndash, 48 months). We assessed the infants&rsquo, gross motor development, fine motor development, language, and personal&ndash, social skills using a modified version of Malawi Development Assessment Tool. The initial clinical trial registration number was NCT03523182. Children in the SP group had higher scores in gross and fine motor development, language, and social skills than those in the CON group. Home fortification of complementary foods using spirulina during the first 1000 days improved development among Zambian children at preschool age.

In vitro M cell models, consisting of co-cultures of Caco-2 cells and lymphoid cells, were developed and examined to observe bacterial transport. However, under our experimental conditions, the differentiation of Caco-2 cells into M cell-like cells could not be induced efficiently. To obtain a functionally stable M cell model based on human cells, C2BBe1 cells were screened and co-cultured with human Raji cells. In our co-cultures, increased sialyl Lewis A antigen expression and decreased Ulex europeaus agglutinin 1 binding were observed. Regarding the functional properties of the model, microsphere and lactic acid bacteria transport across the C2BBe1 co-cultures were increased compared with the levels seen in monocultures. The C2BBe1 monolayers that were co-cultured with Raji cells exhibited some M cell features; therefore, we consider our M cell model to be useful for investigating the interactions of bacteria with M cells.

A laser-plasma source for extreme-ultraviolet (EUV) light that uses a rotating cryogenic solid-state Xe target has been characterized. We focused on parameters at the wavelength of 13.5 nm with 2% bandwidth required for an EUV lithography source and investigated improvements of the conversion efficiency (CE). With the drum rotating, there was an increase in CE and less fast ions compared with the case for the drum at rest. It is considered that the Xe gas on the target surface can produce optimal-scale plasma, and satellite emission lines in Xe plasma effectively increase the EUV intensity, and the ion number is decreased by the gas curtain effect. The dependence of CE on the laser wavelength, laser energy and intensity also studied. As a result, the maximum CE was 0.9% at 13.5 nm with 2% bandwidth under the optimal condition. By continuous irradiation of a Nd:YAG slab laser at a repetition rate of 320 Hz and an average power of 110 W, the target continuously generated EUV light with an average power of 1 W at 13.5 nm with 2% bandwidth. The achieved performances provide valuable information for the design of a future EUV lithography source.

Toll-like receptor (TLR) signals perform a crucial role in innate immune responses to pathogens. In this study, we found that the aryl hydrocarbon receptor (Ahr) negatively regulates inflammatory responses mediated by lipopolysaccharide (LPS) in macrophages. Ahr was induced in macrophages stimulated by LPS, but not by transforming growth factor (TGF)-β plus interleukin (IL)-6, which can induce Ahr in naive T cells. The production of IL-6 and tumor necrosis factor (TNF)-α by LPS was significantly elevated in Ahr-deficient macrophages compared with that in wild-type (WT) cells. Ahr-deficient mice were more highly sensitive to LPS-induced lethal shock than WT mice. Signal transducer and activator of transcription 1 (Stat1) deficiency, as well as Ahr deficiency, augmented LPS-induced IL-6 production. We found that Ahr forms a complex with Stat1 and nuclear factor-kappa B (NF-κB) in macrophages stimulated by LPS, which leads to inhibition of the promoter activity of IL-6. Ahr thus plays an essential role in the negative regulation of the LPS signaling pathway through interaction with Stat1.

Background Most lactic acid bacteria are non-motile but some of them are flagellated and exhibit motility. So far, motile lactobacilli have rarely been studied, and characteristics of their flagellins are poorly understood. In this study, a highly motile strain of Lactobacillus agilis was recruited for transcriptional analysis and characterization of its flagellins. Results Unlike another motile lactic acid bacteria of intestinal isolate, Lactobacillus ruminis, flagellar filaments of the L. agilis strain probably consist of two homologous but distinct flagellins. Glycosylation of the flagellar filaments and their resistance to heat, acid and SDS were also observed. The immunological activity of the flagellins was evaluated through the stimulation of Caco-2 cells. The results show that TLR5-stimulating activity of the protein is attenuated, likely due to an incomplete TLR5-recognition site. Conclusions The flagella filaments of L. agilis BKN88 consist of two homologous glycosylated flagellins, which likely have an incomplete TLR5-recognition site. The characteristics of the flagellin are presumably a consequence of adaptation as a commensal microbe in the gastrointestinal tract.

Interleukin-6 (IL-6) is a prototypical cytokine with functional pleiotropy and plays an important role in host defense. When infections or tissue injuries occur, IL-6 is promptly produced by monocytes and macrophages and contributes to removal of infectious agents and restoration of damaged tissues through activation of immune, hematological, and acute-phase responses. Once stress is removed from the host, IL-6 synthesis ends, but uncontrolled excessive or persistent IL-6 production plays a pathological role in the development of various inflammatory diseases and cancers, indicating that IL-6 is a double-edged sword for the host. Thus, the proper IL-6 expression is very important for host defense and is strictly controlled by chromatin structure, transcriptional regulation, and posttranscriptional modification. Differentiation status of cells, various transcription factors, RNA-binding proteins, and microRNAs are involved in this process. Since it is assumed that dysregulation of any of these regulatory molecules may cause abnormal IL-6 expression in a particular disease, further elucidation of the factors and processes involved in IL-6 expression can be expected to facilitate to clarification of pathogenesis and to identification of novel target molecule(s) for specific diseases.