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Extended pluripotent stem cells (EPSCs) derived from mice and humans showed an enhanced potential for chimeric formation. By exploiting transcriptomic approaches, we assessed the differences in gene expression profile between extended EPSCs derived from mice and humans, and those newly derived from the common marmoset (marmoset; Callithrix jacchus). Although the marmoset EPSC-like cells displayed a unique colony morphology distinct from murine and human EPSCs, they displayed a pluripotent state akin to embryonic stem cells (ESCs), as confirmed by gene expression and immunocytochemical analyses of pluripotency markers and three-germ-layer differentiation assay. Importantly, the marmoset EPSC-like cells showed interspecies chimeric contribution to mouse embryos, such as E6.5 blastocysts in vitro and E6.5 epiblasts in vivo in mouse development. Also, we discovered that the perturbation of gene expression of the marmoset EPSC-like cells from the original ESCs resembled that of human EPSCs. Taken together, our multiple analyses evaluated the efficacy of the method for the derivation of marmoset EPSCs. This article is protected by copyright. All rights reserved.

Summary Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia., Graphical abstract, Highlights • A species-wide method for deriving transgene-free iPSCs has been established • Primary colony-forming cells showed a neural stem cell-like profile, named iNSLCs • mRNA-seq was performed for dissecting the difference between iNSLCs and iPSCs • Single-cell analysis was performed for assessing the heterogeneity of iNSLCs, In this article, Okano and colleagues established a species-wide method for deriving transgene-free iPSCs, and discovered primary colony-forming cells showed a neural stem cell-like profile, named induced neural stem cell-like cells (iNSLCs).

A boid is a simple multiagent model of animal group behavior. Boid agents communicate locally. I studied a heterogeneous boid model comprised of many agents that are divided into several types. While varying interaction among types of agents, this model generates stable patterns with a symmetric interface among different types of agents. As the number of agents increases and as agent clusters grow larger, this model forms a metastable pattern (i.e., a complex of such stable patterns) that is caused by conflict among the local growths of stable patterns. To avoid metastable patterns, I designed two extended heterogeneous boid models: a two-component boid with noise control and a three-component boid with a type transition of agents. In this paper, I examined how these extended models rearrange agents from metastable patterns into stable patterns. These extended models generate stable patterns regardless of the agent number in a shorter time than the original heterogeneous boid model.

Neuronal circuits are shaped by experience during time windows of increased plasticity in postnatal development. In the auditory system, the critical period for the simplest sounds—pure frequency tones—is well defined. Critical periods for more complex sounds remain to be elucidated. We used in vivo electrophysiological recordings in the mouse auditory cortex to demonstrate that passive exposure to frequency modulated sweeps (FMS) from postnatal day 31 to 38 leads to long-term changes in the temporal representation of sweep directions. Immunohistochemical analysis revealed a decreased percentage of layer 4 parvalbumin-positive (PV+) cells during this critical period, paralleled with a transient increase in responses to FMS, but not to pure tones. Preventing the PV+ cell decrease with continuous white noise exposure delayed the critical period onset, suggesting a reduction in inhibition as a mechanism for this plasticity. Our findings shed new light on the dependence of plastic windows on stimulus complexity that persistently sculpt the functional organization of the auditory cortex.

Background The association between delivery mode and subsequent development of diseases is a growing area of research. Cesarean delivery affects the diversity of the microbiota in the infant gut, which may be associated with gastrointestinal disorders, including functional constipation, in infants. In this study, we investigated the association between delivery mode and prevalence of functional constipation in 3-year-old Japanese toddlers. Methods This study used data from the Japan Environment and Children’s Study, an ongoing nationwide birth cohort study. We analyzed 71,878 toddler–mother pairs. The presence of functional constipation was determined according to the Rome III diagnostic criteria. Odds ratios and 95% confidence intervals were calculated using logistic regression analysis. Results The prevalence of functional constipation in 3-year-old Japanese toddlers was estimated to be 12.3%. Logistic regression analysis revealed that the prevalence of functional constipation was higher in toddlers born by cesarean delivery (13.1%) compared with those born by vaginal delivery (12.1%), independent of 22 confounders (adjusted odds ratios = 1.064, 95% confidence interval = 1.004–1.128). Conclusions We determined the prevalence of functional constipation in 3-year-old Japanese toddlers and found that delivery mode was associated with the prevalence of functional constipation in Japanese toddlers.

Identifying the cells from which cancers arise, and the paths they take towards malignancy, is critical for understanding the molecular basis of tumor initiation and progression. To understand if stem and progenitor cells can serve as cells of origin for cancer, we created a model in which the CreERT2 recombinase was knocked into the endogenous locus of the stem cell determinant Msi2. When crossed to a conditional CAG-LSL-MycT58A model, Msi2-CreERT2 mice developed a diversity of tumors across tissues, including multiple pancreatic cancer subtypes: pancreatic ductal adenocarcinoma (PDAC), adenosquamous carcinoma of the pancreas (ASCP), acinar cell carcinoma (ACC), and rare anaplastic tumors. Using single cell analysis, we traced the temporal changes that occurred as normal Msi2+ cells evolved through a pre-cancerous stage to PDAC, ASCP or ACC. These results revealed that Msi2+ cells were present predominantly in pancreatic ducts and exhibited heterogeneous differentiation states in the normal pancreas. At initiation, Myc triggered oncogenic transformation in the most undifferentiated subset leading to the rise of pre-cancer cells with multi-lineage properties. Subsequently, these pre-cancer cells were driven along different fates by epigenetically activated transcriptional programs with genomic changes amplifying the progression into distinct pancreatic cancer subtypes. Finally, integrating transcriptomic and functional genomic approaches in this new model allowed us to define Ifne, Ifitm3, Atf3, Hmmr, and Raet1e as novel functional dependencies of ASCP, giving us unique insight into the most lethal of pancreatic malignancies. These data show that multiple pancreatic cancer subtypes can arise from a common pool of pre-malignant cells and provide a powerful molecular framework to understand the programs that shape divergent fates in pancreas cancer and develop approaches for early detection and interception.

Objective The physiological expression of cell division cycle 42 (cdc42) in major salivary glands, and paracellular transport of fluorescein isothiocyanate-dextran (FITC-dextran) in SMIE cells, which regulate cdc42 expression, was investigated to clarify the involvement of cdc42 in salivary production. Design The physiological expression of cdc42 in the rat submandibular gland, parotid gland, sublingual gland, and SMIE cells was detected using SDS-PAGE and western blotting. The paracellular transport of FITC-dextran in transwells was compared in transfected SMIE cells, exhibiting up- or downregulated cdc42 expression. Results Cdc42 was expressed in all major salivary glands and SMIE cells. SMIE cells transfected with the cdc42 plasmid had an increase efflux. In addition, SMIE cells transfected with the cdc42 siRNA showed decreased efflux. Conclusion We suggest that cdc42 enhances paracellular transport in salivary glands without any morphological changes, including cell-cell adhesion.

Patients with advanced heart failure often experience dyspnea, fatigue, edema, and appetite loss. If these symptoms are refractory to treatment, palliative care via a team approach is necessary. We describe a patient with stage D heart failure whose dyspnea and overall condition improved with comprehensive medical treatments including conventional medications for heart failure, continuous infusions of catecholamine and diuretic, and oral hydromorphone. A 67-year-old man with a 12-year history of dilated cardiomyopathy was admitted to our hospital due to exacerbation of heart failure. Despite continuous infusion of catecholamine and diuretic, his dyspnea and liver and renal function continued to worsen. Oral hydromorphone was administered to relieve his refractory dyspnea, which also improved his conditions, continuous infusion of the catecholamine and diuretic could withdraw. Oral low-dose hydromorphone used in the present case might be a helpful agent for treating dyspnea in stage D heart failure patients with renal dysfunction.

What is ‘relational theorizing’ in International Relations and what can it offer? This article introduces a thematic section that responds to these questions by showing two things. First, relational theorizing is not a doctrine or a method, but a set of analyses that begin with relations rather than the putative essences of constitutively autonomous actors. Second, relational theorizing has emerged from different geo-linguistic traditions, and a relational approach to International Relations (IR) can offer the language and space for increased and productive engagement beyond Anglophone scholarship. This thematic section takes a significant step in this direction by staging a dialogue between Sinophone and Anglophone scholarship on relational IR theorizing. Such an engagement shows points of comparison and contrast, convergence and divergence. In this way, the essays presented here contribute to developing a more ‘global’ IR.

Summary Mutations in the microtubule-associated protein tau (MAPT) gene are known to cause familial frontotemporal dementia (FTD). The R406W tau mutation is a unique missense mutation whose patients have been reported to exhibit Alzheimer’s disease (AD)-like phenotypes rather than the more typical FTD phenotypes. In this study, we established patient-derived induced pluripotent stem cell (iPSC) models to investigate the disease pathology induced by the R406W mutation. We generated iPSCs from patients and established isogenic lines using CRISPR/Cas9. The iPSCs were induced into cerebral organoids, which were dissociated into cortical neurons with high purity. In this neuronal culture, the mutant tau protein exhibited reduced phosphorylation levels and was increasingly fragmented by calpain. Furthermore, the mutant tau protein was mislocalized and the axons of the patient-derived neurons displayed morphological and functional abnormalities, which were rescued by microtubule stabilization. The findings of our study provide mechanistic insight into tau pathology and a potential for therapeutic intervention., Graphical Abstract, Highlights • Phenotype analysis of FTD R406W mutant tau using iPSC-derived neurons • Homogeneous neuronal population obtained via dissociation of cerebral organoids • Altered phosphorylation and fragmentation state of R406W mutant tau • Multiple axonal defects in patient neurons rescued with microtubule stabilization, In this article, Nakamura and colleagues establish an iPSC-derived neuronal model from frontotemporal dementia patients with the tau R406W mutation and gain insight into the disease pathology. Here, the R406W mutant tau exhibited reduced phosphorylation levels and was prone to fragmentation by calpain. Furthermore, the patients’ neurons displayed multiple axonal defects caused by microtubule destabilization.

Narrative, we argue, can (re)construct social reality. Alternative imaginaries of ‘being in the world’ can lead to alternative ways of ‘doing in the world’. We discuss the current dispute between China and Japan over the Senkaku/Diaoyu Islands as an example. Westphalian logic would have the two countries come to blows, if not go to war, over the Islands. The Westphalian account of the dispute centres on the key principle of sovereignty. But what if we utilized a different imaginary to re-narrate the conflict? We turn to popular culture in both Japan and China as a guide, and juxtapose two anime, Appleseed and Time of Eve, with one Chinese TV drama, Nirvana in Fire. Each of these upends conventional analyses of the Islands dispute and offers alternative conceptions of sovereignty. We conclude by considering the implications of such alternative imaginaries for the study, if not practice, of international relations.

Introduction We previously reported that LP positive patients after percutaneous coronary intervention (PCI) had higher rate of re-hospitalization in the small-scale study (135 patients). In this study, we evaluated correlation between LP and later cardiac events leading to re-hospitalization more extensively in greater population. Methods and results A 24-h high-resolution (HR) ambulatory electrocardiogram (ECG) was performed in 421 patients that received PCI for the treatment of acute coronary syndrome (ACS) within 30 days. Various baseline characteristics and post-PCI ECG parameters including LP were examined for correlation with later re-hospitalization. LP was evaluated based on 3 different conditions, i.e., the worst, mean and best values, from 24-h signal-averaged QRS wave data. During the post-PCI follow-up period (611 ± 489.0 days), 90 patients were re-hospitalized due to cardiac events. Multivariate analysis identified only positive LP based on the worst value as an independent predictor for re-hospitalization with OR 2.26. Most of re-hospitalization cases (>75%) were predominantly attributed to ischemic events. LP positive population had significantly higher incidences of ischemic events as well as overall re-hospitalization compared to LP negative population. The predictive power of LP was decreased when it was combined with other variables. The receiver operating characteristic analysis determined the LP cut-off values consistent with the LP positive criteria previously reported and standardized. Conclusion The presence of LP in the 24-h HR ambulatory ECG post-PCI was an independent predictor for a risk of re-hospitalization due to ischemic cardiac events in ACS patients.

Previously, we reported that intravenous injection of chondroitin sulfate (CS), followed by intravenous injection of cationic liposome/siRNA complex (cationic lipoplex) could deliver siRNA into the liver. In this study, we examined the effects of the type of cationic lipid in the liposome on siRNA delivery into the liver. We used 6 types of cationic cholesterol derivatives and 11 types of dialkyl or trialkyl cationic lipids, and prepared 17 types of cationic liposomes to evaluate the gene-silencing effect of siRNA in mouse liver following sequential injection of CS plus cationic lipoplex. In 14 types of cationic liposomes, siRNA was accumulated in the liver when cationic lipoplexes were injected immediately after CS. However, suppression of apolipoprotein B (ApoB) mRNA expression in the liver and reduction of low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) cholesterol level in the serum were observed only after sequential injection of CS plus ApoB siRNA lipoplex with cationic liposomes composed of N-hexadecyl-N,N-dimethylhexadecan-1-aminium bromide (DC-1-16)/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and N,N-dimethyl-N-octadecyloctadecan-1-aminium bromide (DC-1-18)/DOPE. Based on these findings, the type of cationic lipid strongly affected the gene-silencing effect of siRNA in the liver by sequential injection of CS plus cationic lipoplex.

Background It is unclear how acute coronavirus disease 2019 (COVID-19)-directed therapies are used in children with life-threatening COVID-19 in US hospitals. We described characteristics of children hospitalized in the intensive care unit or step-down unit (ICU/SDU) who received COVID-19-directed therapies and the specific therapies administered. Methods Between March 15, 2020 and December 27, 2020, children, No therapies for acute coronavirus disease 2019 (COVID-19) have been tested in children to establish effectiveness. Despite this, across 48 hospitals, 235/424 children (55%) admitted to the intensive care or step-down unit with COVID-19 received directed therapies. Systemic steroids and remdesivir were most commonly administered, with use rising from March 2020 to December 2020.

IntroductionGiven an increasing use of dipeptidyl peptidase-4 (DPP-4) inhibitors to treat patients with type 2 diabetes mellitus in the real-world setting, we conducted a prospective observational study (Japan-based Clinical Research Network for Diabetes Registry: J-BRAND Registry) to elucidate the safety and efficacy profile of long-term usage of alogliptin.Research design and methodsWe registered 5969 patients from April 2012 through September 2014, who started receiving alogliptin (group A) or other classes of oral hypoglycemic agents (OHAs; group B), and were followed for 3 years at 239 sites nationwide. Safety was the primary outcome. Symptomatic hypoglycemia, pancreatitis, skin disorders of non-extrinsic origin, severe infections, and cancer were collected as major adverse events (AEs). Efficacy assessment was the secondary outcome and included changes in hemoglobin A1c (HbA1c), fasting blood glucose, fasting insulin and urinary albumin.ResultsOf the registered, 5150 (group A: 3395 and group B: 1755) and 5096 (3358 and 1738) were included for safety and efficacy analysis, respectively. Group A patients mostly (>90%) continued to use alogliptin. In group B, biguanides were the primary agents, while DPP-4 inhibitors were added in up to ~36% of patients. The overall incidence of AEs was similar between the two groups (42.7% vs 42.2%). Kaplan-Meier analysis revealed the incidence of cancer was significantly higher in group A than in group B (7.4% vs 4.8%, p=0.040), while no significant incidence difference was observed in the individual cancer. Multivariate Cox regression analysis revealed that the imbalanced patient distribution (more elderly patients in group A than in group B), but not alogliptin usage per se, contributed to cancer development. The incidence of other major AE categories was with no between-group difference. Between-group difference was not detected, either, in the incidence of microvascular and macrovascular complications. HbA1c and fasting glucose decreased significantly at the 0.5-year visit and nearly plateaued thereafter in both groups.ConclusionsAlogliptin as a representative of DPP-4 inhibitors was safe and durably efficacious when used alone or with other OHAs for patients with type 2 diabetes in the real world setting.

BLIMP1 (PRDM1) and VASA (DDX4) play pivotal roles in the development of the germ cell linage. Importantly, these genes are specifically expressed in germ cells; BLIMP1 in primordial germ cells (PGCs) to early-stage gonocytes, and VASA in migration-stage PGCs to mature gametes. The high reproductive efficiency of common marmosets (marmosets; Callithrix jacchus) makes them advantageous for use in germ cell research. We herein report the generation of a male marmoset embryonic stem cell (ESC) line harboring BLIMP1 and DDX4 double reporters. This ESC line will be a useful tool for investigating male gametogenesis in non-human primates.

Mammalian pluripotent stem cells are thought to exist in two states: naive and primed. Generally, unlike those in rodents, pluripotent stem cells in primates, including humans, are regarded as being in the primed pluripotent state. Recently, several groups reported the existence of naive pluripotent stem cells in humans. In this study, we report the conversion of primed state embryonic stem cells from common marmoset, a New World monkey, to the naive state using transgenes. The cells showed typical naive state features, including dome-like colony morphology, growth factor requirement, gene expression profile, X chromosome activation state, and energy metabolic status. Moreover, interspecies chimeric embryo formation ability with mouse embryos was increased in the naive state. This technique can be applied in basic medical research using nonhuman primates, such as preclinical use of naive pluripotent stem cells and generating genetically modified primates.

Obtaining differentiated cells with high physiological functions by an efficient, but simple and rapid differentiation method is crucial for modeling neuronal diseases in vitro using human pluripotent stem cells (hPSCs). Currently, methods involving the transient expression of one or a couple of transcription factors have been established as techniques for inducing neuronal differentiation in a rapid, single step. It has also been reported that microRNAs can function as reprogramming effectors for directly reprogramming human dermal fibroblasts to neurons. In this study, we tested the effect of adding neuronal microRNAs, miRNA-9/9*, and miR-124 (miR-9/9*-124), for the neuronal induction method of hPSCs using Tet-On-driven expression of the Neurogenin2 gene (Ngn2), a proneural factor. While it has been established that Ngn2 can facilitate differentiation from pluripotent stem cells into neurons with high purity due to its neurogenic effect, a long or indefinite time is required for neuronal maturation with Ngn2 misexpression alone. With the present method, the cells maintained a high neuronal differentiation rate while exhibiting increased gene expression of neuronal maturation markers, spontaneous calcium oscillation, and high electrical activity with network bursts as assessed by a multipoint electrode system. Moreover, when applying this method to iPSCs from Alzheimer's disease (AD) patients with presenilin-1 (PS1) or presenilin-2 (PS2) mutations, cellular phenotypes such as increased amount of extracellular secretion of amyloid &beta, 42, abnormal oxygen consumption, and increased reactive oxygen species in the cells were observed in a shorter culture period than those previously reported. Therefore, it is strongly anticipated that the induction method combining Ngn2 and miR-9/9*-124 will enable more rapid and simple screening for various types of neuronal disease phenotypes and promote drug discovery.

Summary Several groups have reported the existence of a form of pluripotency that resembles that of mouse embryonic stem cells (mESCs), i.e., a naive state, in human pluripotent stem cells; however, the characteristics vary between reports. The nuclear receptor ESRRB is expressed in mESCs and plays a significant role in their self-renewal, but its expression has not been observed in most naive-like human induced pluripotent stem cells (hiPSCs). In this study, we modified several methods for converting hiPSCs into a naive state through the transgenic expression of several reprogramming factors. The resulting cells express the components of the core transcriptional network of mESCs, including ESRRB, at high levels, which suggests the existence of naive-state hiPSCs that are similar to mESCs. We also demonstrate that these cells differentiate more readily into neural cells than do conventional hiPSCs. These features may be beneficial for their use in disease modeling and regenerative medicine., Graphical Abstract, Highlights • Induction methods for human naive pluripotent stem cells were optimized • hiPSCs were reprogrammed to a naive-like state with six transcription factors • The resulting naive-like hiPSCs expressed naive state-related genes, including ESRRB • The naive-like hiPSCs differentiated readily into neural and glial cells, Kisa et al. modified several methods for converting human induced pluripotent stem cells (hiPSCs) into a naive state, a form of pluripotency that exists in mouse embryonic stem cells (ESCs). Converted cells express components of the core transcriptional network upregulated in mouse ESCs, including ESRRB. They also show that these cells differentiate more readily into neural cells than do conventional hiPSCs.