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Human‐induced pluripotent stem cells (hiPSCs) have great therapeutic potential. The cell source differentiated from hiPSCs requires xeno‐free and robust methods for lineage‐specific differentiation. Here, a system is described for differentiating hiPSCs on new generation laminin fragments (NGLFs), a recombinant form of a laminin E8 fragment conjugated to the heparan sulfate chains (HS) attachment domain of perlecan. Using NGLFs, hiPSCs are highly promoted to direct differentiation into a paraxial mesoderm state with high‐efficiency muscle lineage generation. HS conjugation to the C‐terminus of Laminin E8 fragments brings fibroblast growth factors (FGFs) bound to the HS close to the cell surface of hiPSCs, thereby facilitating stronger FGF signaling pathways stimulation and initiating HOX gene expression, which triggers the paraxial mesoderm differentiation of hiPSCs. This highly efficient differentiation system can provide a roadmap for paraxial mesoderm development and an infinite source of myocytes and muscle stem cells for disease modeling and regenerative medicine. [ABSTRACT FROM AUTHOR]

Rotaxanes consisting of a high‐molecular‐weight axle and wheel components (macro‐rotaxanes) have high structural freedom, and are attractive for soft‐material applications. However, their synthesis remains underexplored. Here, we investigated macro‐rotaxane formation by the topological trapping of multicyclic polydimethylsiloxanes (mc‐PDMSs) in silicone networks. mc‐PDMS with different numbers of cyclic units and ring sizes was synthesized by cyclopolymerization of a α,ω‐norbornenyl‐functionalized PDMS. Silicone networks were prepared in the presence of 10–60 wt % mc‐PDMS, and the trapping efficiency of mc‐PDMS was determined. In contrast to monocyclic PDMS, mc‐PDMSs with more cyclic units and larger ring sizes can be quantitatively trapped in the network as macro‐rotaxanes. The damping performance of a 60 wt % mc‐PDMS‐blended silicone network was evaluated, revealing a higher tan δ value than the bare PDMS network. Thus, macro‐rotaxanes are promising as non‐leaching additives for network polymers. [ABSTRACT FROM AUTHOR]

The outbreak‐causing monkeypox virus of 2022 (2022 MPXV) is classified as a clade IIb strain and phylogenetically distinct from prior endemic MPXV strains (clades I or IIa), suggesting that its virological properties may also differ. Here, we used human keratinocytes and induced pluripotent stem cell‐derived colon organoids to examine the efficiency of viral growth in these cells and the MPXV infection‐mediated host responses. MPXV replication was much more productive in keratinocytes than in colon organoids. We observed that MPXV infections, regardless of strain, caused cellular dysfunction and mitochondrial damage in keratinocytes. Notably, a significant increase in the expression of hypoxia‐related genes was observed specifically in 2022 MPXV‐infected keratinocytes. Our comparison of virological features between 2022 MPXV and prior endemic MPXV strains revealed signaling pathways potentially involved with the cellular damages caused by MPXV infections and highlights host vulnerabilities that could be utilized as protective therapeutic strategies against human mpox in the future. [ABSTRACT FROM AUTHOR]

Human in vitro oogenesis provides a framework for clarifying the mechanism of human oogenesis. To create its benchmark, it is vital to promote in vitro oogenesis using a model physiologically close to humans. Here, we establish a foundation for in vitro oogenesis in cynomolgus (cy) monkeys (Macaca fascicularis): cy female embryonic stem cells harboring one active and one inactive X chromosome (Xa and Xi, respectively) differentiate robustly into primordial germ cell‐like cells, which in xenogeneic reconstituted ovaries develop efficiently into oogonia and, remarkably, further into meiotic oocytes at the zygotene stage. This differentiation entails comprehensive epigenetic reprogramming, including Xi reprogramming, yet Xa and Xi remain epigenetically asymmetric with, as partly observed in vivo, incomplete Xi reactivation. In humans and monkeys, the Xi epigenome in pluripotent stem cells functions as an Xi‐reprogramming determinant. We further show that developmental pathway over‐activations with suboptimal up‐regulation of relevant meiotic genes impede in vitro meiotic progression. Cy in vitro oogenesis exhibits critical homology with the human system, including with respect to bottlenecks, providing a salient model for advancing human in vitro oogenesis. Synopsis: This study establishes a foundation for in vitro oogenesis in cynomolgus monkeys (Macaca fascicularis) by demonstrating the induction of fetal meiotic oocyte‐like cells from embryonic stem cells. Monkey in vitro oogenesis exhibits critical homology with the human system, providing a salient model for advancing human in vitro oogenesis.Induction of fetal meiotic oocyte‐like cells from ESCs in cynomolgus monkeys.Oocyte‐like cell development entails nearly complete DNA methylation reprogramming.X‐chromosome reactivation is partial with remaining epigenetic asymmetries between two X chromosomes.Oocyte‐like cells over‐activate developmental genes with suboptimal meiotic gene expression. [ABSTRACT FROM AUTHOR]

Background: Hypoglycemia is a significant problem for all neonates and requires minimally invasive and reliable monitoring. The primary objective of this study was to verify the safety and accuracy of the continuous glucose monitoring (CGM) of full‐term neonates using Freestyle Libre, a flash glucose monitoring (FGM) device. Methods: The study was conducted on 20 neonates. Shortly after birth, we placed the FGM sensor on the outside of the neonates' thighs. We scanned the CGM values at 60, 120, 180, and 360 min after birth and simultaneously obtained blood glucose values with plantar capillaries by heel puncture. The neonates wore the sensors for up to 6 h and then they were removed. Results: Of the 75 data points to be measured, 65 points (86.7%) were obtained by scan. There was no change in the sensor attachment site in 12 of 18 completed cases in this study but we observed slight induration in four cases (22.2%) and slight redness in one case (5.5%) at the sensor puncture site. A moderate correlation was observed between the CGM and blood glucose values. The CGM values tended to be low at 120, 180, and 360 min after birth, and tended to be high only at 60 min after birth. Conclusions: The CGM device was safe to wear on the neonate and the CGM data correlated well with blood glucose levels. There was dissociation between CGM data and blood glucose levels in the acute period soon after birth when the blood glucose levels changed rapidly. [ABSTRACT FROM AUTHOR]

Early diagnosis of pancreatic ductal adenocarcinoma (PDAC) is essential for improving prognosis; however, diagnosing PDAC at an early stage is challenging. In patients with localized high‐grade pancreatic intraepithelial neoplasia (HG‐PanIN), whose tumorous lesion is undetectable on cross‐sectional images such as computed tomography or magnetic resonance image, long‐term survival is expected. Pancreatic cystic lesions or main pancreatic duct (MPD) dilatation are important indirect findings for the initial diagnosis of HG‐PanIN. Magnetic resonance cholangiopancreatography (MRCP) and endoscopic ultrasound (EUS) should play important roles in detecting abnormal image findings, such as local irregular MPD stenosis, caliber MPD changes, small cystic lesions, or branch duct dilatation. Additionally, EUS could detect hypoechoic areas around the MPD stenosis in some patients with HG‐PanIN. Subsequently, endoscopic retrograde cholangiopancreatography (ERCP) and its associated pancreatic juice cytology, including serial pancreatic juice aspiration cytologic examination (SPACE) after placement of an endoscopic nasopancreatic drainage (ENPD) tube, may have high diagnostic accuracy for confirming the malignancy in HG‐PanIN. Although ERCP and its associated pancreatic cytology, including SPACE, may be associated with post‐ERCP pancreatitis (PEP), a recent randomized trial suggested that a 4‐Fr ENPD tube may reduce the incidence of PEP. In the future, further prospective multicenter studies are required to establish a standard method of SPACE. Additionally, further studies for novel biomarkers could help to establish evolutionary methods with duodenal fluid and pancreatic juice for the early and accurate diagnosis of early‐stage PDAC. [ABSTRACT FROM AUTHOR]

In pregnant mice, the maternal liver expands drastically during gestation, which is believed to be essential to accommodate various metabolic demands caused by physiological changes and fetal growth. Although hepatocyte proliferation and hypertrophy have been reported, little is known about the dynamics of biliary epithelial cells (BECs), which comprise the bile duct epithelium in the liver. Here, we show that BECs transiently proliferate during the early stage of gestation. Lineage tracing revealed that BEC progeny were retained in the bile duct epithelium and did not differentiate into hepatocytes, indicating BEC self‐replication during pregnancy. RNA‐sequencing analysis of BECs identified their early pregnancy‐signature transcriptomes, which highlighted Yes‐associated protein (YAP) signaling‐related genes. Nuclear accumulation of YAP was enhanced in BECs during pregnancy but was barely detectable in hepatocytes. In addition, the pharmacological inhibition of YAP attenuated BEC proliferation and liver weight gain during pregnancy. Our results delineate the proliferation and transcriptomic dynamics of BECs during pregnancy and suggest the relevance of YAP‐mediated signals. [ABSTRACT FROM AUTHOR]

This 31‐parameter panel was developed for simultaneously measuring multiple immune cell populations including T cells, B cells, natural killer cells, dendritic cells, monocytes, and hematopoietic progenitor cells in human peripheral blood mononuclear cells. This panel enables the capture of individual immune dynamics and assessments of single‐cell changes in the immune system that are associated with aging and diseases. This panel includes markers to separate the differentiation status of each cell population and might be applicable to studies of infectious and autoimmune diseases, as patient samples are usually limited in volume and require an analysis system that provides a relatively large amount of information. [ABSTRACT FROM AUTHOR]

The purification step in the manufacturing of cyclic polymers is difficult as complete fractionation to eliminate linear impurities requires considerable effort. Here, we report a new polymer separation methodology that uses metal‐organic frameworks (MOFs) to discriminate between linear and cyclic polyethylene glycols (PEGs) via selective polymer insertion into the MOF nanopores. Preparation of a MOF‐packed column allowed analytical and preparative chromatographic separation of these topologically distinct pairs. In addition, gram‐scale PEGs with only cyclic structures were successfully obtained from a crude reaction mixture by using MOF as an adsorbent. [ABSTRACT FROM AUTHOR]

Human germ cells perpetuate human genetic and epigenetic information. However, the underlying mechanism remains elusive, due to a lack of appropriate experimental systems. Here, we show that human primordial germ cell‐like cells (hPGCLCs) derived from human‐induced pluripotent stem cells (hiPSCs) can be propagated to at least ~106‐fold over a period of 4 months under a defined condition in vitro. During expansion, hPGCLCs maintain an early hPGC‐like transcriptome and preserve their genome‐wide DNA methylation profiles, most likely due to retention of maintenance DNA methyltransferase activity. These characteristics contrast starkly with those of mouse PGCLCs, which, under an analogous condition, show a limited propagation (up to ~50‐fold) and persist only around 1 week, yet undergo cell‐autonomous genome‐wide DNA demethylation. Importantly, upon aggregation culture with mouse embryonic ovarian somatic cells in xenogeneic‐reconstituted ovaries, expanded hPGCLCs initiate genome‐wide DNA demethylation and differentiate into oogonia/gonocyte‐like cells, demonstrating their germline potential. By creating a paradigm for hPGCLC expansion, our study uncovers critical divergences in expansion potential and the mechanism for epigenetic reprogramming between the human and mouse germ cell lineage. Synopsis: In vitro reconstitution of germ‐cell development is a key for analyzing and enhancing reproductive biology. This resource reports a culture method for robust propagation of human primordial germ cell‐like cells (hPGCLCs) under defined conditions, establishing a tractable paradigm for human germ‐cell expansion. Induced pluripotent stem cell‐derived hPGCLCs expand ~106‐fold under forskolin/SCF/bFGF in vitro over a period of four months.During expansion, hPGCLCs maintain an early hPGC(LC) transcriptome and DNA methylome.hPGC(LC) specification and epigenetic reprogramming are genetically dissociable.Expanded hPGCLCs differentiate into oogonia with epigenetic reprogramming in xenogeneic reconstituted ovaries. [ABSTRACT FROM AUTHOR]

Chain‐end‐functionalized poly(3‐hexylthiophene)s (P3HTs) with benzyl alcohol (─PhCH2OH), phenol (─PhOH), and benzoic acid (─PhCOOH) groups are directly synthesized based on the Negishi catalyst‐transfer polycondensation method utilizing the zincate complex of tBu4ZnLi2. In this system, neither protection nor deprotection steps are required, and also providing a living polymerization system to control the molecular weight while maintaining a low molar mass dispersity (ÐM) of the obtained P3HT derivatives. Indeed, the chain‐end‐functionalized P3HTs can be synthesized along with controlled number‐average molecular weights (Mn = 5100–20 000), low ÐM (1.06–1.14), and high chain‐end functionality (Fn = 46–86%). The Fn values for the alcohol and phenol groups are found to be high (86% for ─PhCH2OH and 71% for ─PhOH based on 1H NMR, respectively), as also confirmed by matrix‐assisted laser desorption/ionization time of flight mass spectroscopy. The easily synthesizable chain‐end‐functionalized P3HTs will be applicable for the facile synthesis of block and branched polymers containing P3HT as well as its related semiconducting polymer segments. [ABSTRACT FROM AUTHOR]

Chronic infection with hepatitis B virus (HBV) sometime induces lethal cirrhosis and hepatocellular carcinoma. Although nucleot(s)ide analogs are used as main treatment for HBV infection, the emergence of the drug‐resistant viruses has become a problem. To discover novel antivirals with low side effects and low risk of emergence of resistant viruses, screening for anti‐HBV compounds was performed with compound libraries of inhibitors targeting G‐protein‐coupled receptors (GPCRs). HepG2‐hNTCP C4 cells infected with HBV were treated with various GPCR inhibitors and harvested at 14 day postinfection for quantification of core protein in the first screening or relaxed circular DNA in the second screening. Finally, we identified a cannabinoid receptor 1 inhibitor, rimonabant, as a candidate showing anti‐HBV effect. In HepG2‐hNTCP C4 cells, treatment with rimonabant suppressed HBV propagation at the viral RNA transcription step but had no effect on entry or covalently closed circular DNA level. The values of half maximal inhibitory concentration, half maximal effective concentration, and selectivity index of rimonabant in primary human hepatocyte (PHH) are 2.77 μm, 40.4 μm, and 14.6, respectively. Transcriptome analysis of rimonabant‐treated primary hepatocytes by RNA sequencing revealed that the transcriptional activity of hepatocyte nuclear factor 4α (HNF4α), which is known to stimulate viral RNA synthesis, was depressed. By treatment of PHH with rimonabant, the expression level of HNF4α protein and the production of the messenger RNAs (mRNAs) of downstream factors promoted by HNF4α were reduced while the amount of HNF4α mRNA was not altered. These results suggest that treatment with rimonabant suppresses HBV propagation through the inhibition of HNF4α activity. [ABSTRACT FROM AUTHOR]

Reprogramming technology has enabled the fate conversion of terminally differentiated somatic cells into pluripotent stem cells or into another differentiated state. A dynamic reorganization of epigenetic regulation takes place during cellular reprogramming. Given that reprogramming does not require changes in the underlying genome, the technology can be used to actively modify epigenetic regulation. Although reprogramming has been investigated mostly at the cellular level in vitro, studies have reported that somatic cells are reprogrammable in multicellular organisms in vivo. In vivo reprogramming provides a potential strategy for regenerative medicine. Notably, recent studies using in vivo reprogramming technology to alter epigenetic regulation at organismal levels have revealed unappreciated epigenetic mechanisms in various biological phenomena, including cancer development, tissue regeneration, aging, and rejuvenation in mammals. Moreover, in vivo reprogramming technology can be applied to abrogate epigenetic aberrations associated with aging and cancer, which raises the possibility that the technology could provide a potential strategy to control the fate of detrimental cells such as senescent cells and cancer cells in vivo. Here, we review recent progress and future perspectives of in vivo reprogramming. [ABSTRACT FROM AUTHOR]

The mechanism for sex determination in mammalian germ cells remains unclear. Here, we reconstitute the female sex determination in mouse germ cells in vitro under a defined condition without the use of gonadal somatic cells. We show that retinoic acid ( RA) and its key effector, STRA8, are not sufficient to induce the female germ-cell fate. In contrast, bone morphogenetic protein ( BMP) and RA synergistically induce primordial germ cells ( PGCs)/ PGC-like cells ( PGCLCs) derived from embryonic stem cells ( ESCs) into fetal primary oocytes. The induction is characterized by entry into the meiotic prophase, occurs synchronously and recapitulates cytological and transcriptome progression in vivo faithfully. Importantly, the female germ-cell induction necessitates a proper cellular competence-most typically, DNA demethylation of relevant genes-which is observed in appropriately propagated PGCs/ PGCLCs, but not in PGCs/ PGCLCs immediately after induction. This provides an explanation for the differential function of BMP signaling between PGC specification and female germ-cell induction. Our findings represent a framework for a comprehensive delineation of the sex-determination pathway in mammalian germ cells, including humans. [ABSTRACT FROM AUTHOR]

The expansion of primordial germ cells ( PGCs), the precursors for the oocytes and spermatozoa, is a key challenge in reproductive biology/medicine. Using a chemical screening exploiting PGC-like cells ( PGCLCs) induced from mouse embryonic stem cells ( ESCs), we here identify key signaling pathways critical for PGCLC proliferation. We show that the combinatorial application of Forskolin and Rolipram, which stimulate cAMP signaling via different mechanisms, expands PGCLCs up to ~50-fold in culture. The expanded PGCLCs maintain robust capacity for spermatogenesis, rescuing the fertility of infertile mice. Strikingly, during expansion, PGCLCs comprehensively erase their DNA methylome, including parental imprints, in a manner that precisely recapitulates genome-wide DNA demethylation in gonadal germ cells, while essentially maintaining their identity as sexually uncommitted PGCs, apparently through appropriate histone modifications. By establishing a paradigm for PGCLC expansion, our system reconstitutes the epigenetic 'blank slate' of the germ line, an immediate precursory state for sexually dimorphic differentiation. [ABSTRACT FROM AUTHOR]

Aims/Introduction We devised a new system called 'Educational Guidance' (E-Guide) for nutritional education based on self-efficacy. The present study aimed to examine the effects of E-Guide use on glycemic control among patients with type 2 diabetes. Materials and Methods We carried out an interventional and observational study that included 74 patients with type 2 diabetes. The extent of glycemic control in the 39 patients who received guidance through the E-Guide (E-Guide group) was compared with that of 35 patients who received conventional nutritional guidance (control group). We carried out a 1-year follow-up survey (subanalysis) based on the electronic health records of 18 patients from the E-guide group and 19 patients from the control group. These patients continued treatment at Hikone Municipal Hospital, Hikone, Shiga, Japan. Changes in glycated hemoglobin levels, body mass index and medication dose were examined from time of enrollment to the end of the 1-year intervention, and during the 1-year follow-up. Results Decreases in glycated hemoglobin levels were more pronounced in the E-Guide group than in the control group during the intervention period ( P < 0.05). The levels further decreased during the follow-up period ( P < 0.01). In the E-Guide group, body mass index decreased significantly throughout the follow-up period ( P < 0.001). Additionally, increased medication doses were significantly less common in the E-Guide group than in the control group ( P < 0.01). Conclusions Intervention based on our 'E-Guide' is more useful and powerful than the conventional methods for glycemic control and self-care behavior among patients with type 2 diabetes in Japan. [ABSTRACT FROM AUTHOR]

Aim Given that different hospitals achieve different outcomes, optimal evaluation of treatment outcomes in the local community requires evaluation of many institutions in that area. We carried out a prospective multicenter cohort study in Kagoshima Prefecture to identify factors that contribute to deterioration in activities of daily living performance and patient mortality 1 year after surgical treatment of hip fractures. Methods We prospectively enrolled 387 patients who underwent surgery for hip fractures in 33 registered facilities within a 6-month period from February to July 2007. Logistic regression analysis was carried out to identify factors that contribute to deterioration in activities of daily living performance and death within 1 year after surgery. Results An increased risk of Barthel Index ( BI) deterioration was associated with increased age ( P for trend = 0.003), worse pre-injury BI ( P for trend = 0.021), trochanteric fractures ( OR 2.07, 95% CI 1.31-3.27), worse BI at discharge ( P for trend < 0.001) and postoperatively developed cognitive impairment ( OR 6.34, 95% CI 2.15-18.7). The OR for BI deterioration in patients with newly-diagnosed disease after discharge was approximately 9.16 (95% CI 4.03-20.8). No factors except age and sex were statistically significant as the preoperative indicators of mortality risk. Only BI at discharge was a significant determinant of mortality risk ( P for trend = 0.013) after adjusting for the effects of age and sex. Conclusions Patients with poor activities of daily living performance at the time of hospital discharge were likely to show poor functional recovery and a high 1-year postoperative mortality. Geriatr Gerontol Int 2017; 17: 391-401. [ABSTRACT FROM AUTHOR]

The Activin/Nodal/ TGF-β signaling pathway plays a major role in maintaining mouse epiblast stem cells (Epi SCs). The Epi SC-maintaining medium, which contains Activin A and bFGF, induces differentiation of mouse embryonic stem cells ( ESCs) to Epi SCs. Here, we show that Activin A also has an ability to efficiently propagate ESCs without differentiation to Epi SCs when combined with a MEK inhibitor PD0325901. ESCs cultured in Activin+ PD retained high-level expression of naive pluripotency-related transcription factors. Genomewide analysis showed that the gene expression profile of ESCs cultured in Activin+ PD resembles that of ESCs cultured in 2i. ESCs cultured in Activin+ PD also showed features common to the naive pluripotency of ESCs, including the preferential usage of the Oct4 distal enhancer and the self-renewal response to Wnt pathway activation. Our finding shows a role of Activin/Nodal/ TGF-β signaling in stabilizing self-renewal gene regulatory networks in ESCs. [ABSTRACT FROM AUTHOR]

Three numerical methods, namely, volume of fluid (VOF), simple coupled volume of fluid with level set (S-CLSVOF), and S-CLSVOF with the density-scaled balanced continuum surface force (CSF) model, have been incorporated into OpenFOAM source code and were validated for their accuracy for three cases: (i) an isothermal static case, (ii) isothermal dynamic cases, and (iii) non-isothermal dynamic cases with thermocapillary flow including dynamic interface deformation. Results have shown that the S-CLSVOF method gives accurate results in the test cases with mild computation conditions, and the S-CLSVOF technique with the density-scaled balanced CSF model leads to accurate results in the cases of large interface deformations and large density and viscosity ratios. These show that these high accuracy methods would be appropriate to obtain accurate predictions in multiphase flow systems with thermocapillary flows. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Polyfunctionality and cytotoxic activity dictate CD8+ T‐cell efficacy in the eradication of infected and malignant cells. The induction of these effector functions depends on the specific interaction between the T‐cell receptor (TCR) and its cognate peptide‐MHC class I complex, in addition to signals provided by co‐stimulatory or co‐inhibitory receptors, which can further regulate these functions. Among these receptors, the role of 2B4 is contested, as it has been described as either co‐stimulatory or co‐inhibitory in modulating T‐cell functions. We therefore combined functional, transcriptional and epigenetic approaches to further characterize the impact of disrupting the interaction of 2B4 with its ligand CD48, on the activity of human effector CD8+ T‐cell clones. In this setting, we show that the 2B4‐CD48 axis is involved in the fine‐tuning of CD8+ T‐cell effector function upon antigenic stimulation. Blocking this interaction resulted in reduced CD8+ T‐cell clone‐mediated cytolytic activity, together with a subtle drop in the expression of genes involved in effector function regulation. Our results also imply a variable contribution of the 2B4‐CD48 interaction to the modulation of CD8+ T‐cell functional properties, potentially linked to intrinsic levels of T‐bet expression and TCR avidity. The present study thus provides further insights into the role of the 2B4‐CD48 interaction in the fine regulation of CD8+ T‐cell effector function upon antigenic stimulation. [ABSTRACT FROM AUTHOR]

CpG oligodeoxynucleotide (ODN) is one of promising nucleic acid-based adjuvants. We recently improved its ability to enhance CD8+ T-cell responses to coadministered protein antigen without conjugation or emulsion, by forming a nanoparticulate complex between CpG ODN (K3) and mushroom-derived β-glucan schizophyllan (SPG), namely K3-SPG. Here, we sought to elucidate the cellular immunological mechanisms by which K3-SPG induce such potent CD8+ T-cell responses to coadministered antigen. By focusing on two DC subsets, plasmacytoid DCs and CD8α+ DCs, as well as the secreted cytokines, IFN-α and IL-12, we found that K3-SPG strongly activates mouse plasmacytoid DCs to secrete IFN-α and CD8α+ DCs to secrete IL-12, respectively. Although a single cytokine deficiency had no impact on adjuvant effects, the lack of both type I IFN and IL-12 in mice resulted in a significant reduction of Th1 type immune responses and CD8+ T-cell responses elicited by protein vaccine model. By sharp contrast, type I IFN, but not IL-12, was required for the production of IFN-γ by human PBMCs as well as antigen-specific CD8+ T-cell proliferation. Taken together, K3-SPG may overcome the species barrier for CpG ODN to enhance antigen-specific CD8+ T-cell responses despite the differential role of IL-12 between human and mice. [ABSTRACT FROM AUTHOR]

ABSTRACT The 'topological polymer chemistry' of amphiphilic linear and cyclic block copolymers at an air/water interface was investigated. A cyclic copolymer and two linear copolymers (AB-type diblock and ABA-type triblock copolymers) synthesized from the same monomers were used in this study. Relatively stable monolayers of these three copolymers were observed to form at an air/water interface. Similar condensed-phase temperature-dependent behaviors were observed in surface pressure-area isotherms for these three monolayers. Molecular orientations at the air/water interface for the two linear block copolymers were similar to that of the cyclic block copolymer. Atomic force microscopic observations of transferred films for the three polymer types revealed the formation of monolayers with very similar morphologies at the mesoscopic scale at room temperature and constant compression speed. ABA-type triblock linear copolymers adopted a fiber-like surface morphology via two-dimensional crystallization at low compression speeds. In contrast, the cyclic block copolymer formed a shapeless domain. Temperature-controlled out-of-plane X-ray diffraction (XRD) analysis of Langmuir-Blodgett (LB) films fabricated from both amphiphilic linear and cyclic block copolymers was performed to estimate the layer regularity at higher temperatures. Excellent heat-resistant properties of organized molecular films created from the cyclic copolymer were confirmed. Both copolymer types showed clear diffraction peaks at room temperature, indicating the formation of highly ordered layer structures. However, the layer structures of the linear copolymers gradually disordered when heated. Conversely, the regularity of cyclic copolymer LB multilayers did not change with heating up to 50 °C. Higher-order reflections ( d002, d003) in the XRD patterns were also unchanged, indicative of a highly ordered structure. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2016, 54, 486-498 [ABSTRACT FROM AUTHOR]

A pentacyclic quadruply fused polymer topology has been constructed for the first time through alkyne-azide addition (click) and olefin metathesis (clip) reactions in conjunction with an electrostatic self-assembly and covalent fixation (ESA-CF) process. Thus, a spiro-type, tandem tetracyclic poly(tetrahydrofuran), poly(THF), precursor having two allyloxy groups at the opposite positions of the four ring units was prepared by the click-linking of one unit of an eight-shaped precursor having alkyne groups at the opposite positions with two units of a single-cyclic counterpart having an azide and an alkene group at the opposite positions. Both are obtainable through ESA-CF. The subsequent metathesis clip-folding of the tetracyclic precursor could afford a pentacyclic quadruply fused polymer product, of 'shippo' form, in 19 % yield. [ABSTRACT FROM AUTHOR]

Aberrant activation of the Hedgehog (Hh) pathway has been reported in several malignancies. We previously demonstrated that knockdown of GLI2 inhibited proliferation of osteosarcoma cells through regulation of the cell cycle. In this study, we analyzed the function of GLI2 in the pathogenesis of osteosarcoma metastasis. Immunohistochemical studies showed that GLI2 was overexpressed in patient osteosarcoma specimens. Knockdown of GLI2 inhibited migration and invasion of osteosarcoma cells. In contrast, the forced expression of constitutively active GLI2 in mesenchymal stem cells promoted invasion. In addition, xenograft models showed that knockdown of GLI2 decreased lung metastasis of osteosarcomas. To examine clinical applications, we evaluated the efficacy of arsenic trioxide (ATO), which is a Food and Drug Administration-approved antitumor drug, on osteosarcoma cells. ATO treatment suppressed the invasiveness of osteosarcoma cells by inhibiting the transcriptional activity of GLI2. In addition, the combination of Hh inhibitors including ATO, vismodegib and GANT61 prevented migration and metastasis of osteosarcoma cells. Consequently, our findings suggested that GLI2 regulated metastasis as well as the progression of osteosarcomas. Inhibition of the GLI2 transcription may be an effective therapeutic method for preventing osteosarcoma metastasis. [ABSTRACT FROM AUTHOR]

Nuclear factor-κB ( NF-κB) is a key regulator of cancer progression and the inflammatory effects of disease. To identify inhibitors of DNA binding to NF-κB, we developed a new homogeneous method for detection of sequence-specific DNA-binding proteins. This method, which we refer to as DSE- FRET, is based on two phenomena: protein-dependent blocking of spontaneous DNA strand exchange ( DSE) between partially double-stranded DNA probes, and fluorescence resonance energy transfer ( FRET). If a probe labeled with a fluorophore and quencher is mixed with a non-labeled probe in the absence of a target protein, strand exchange occurs between the probes and results in fluorescence elevation. In contrast, blocking of strand exchange by a target protein results in lower fluorescence intensity. Recombinant human NF-κB (p50) suppressed the fluorescence elevation of a specific probe in a concentration-dependent manner, but had no effect on a non-specific probe. Competitors bearing a NF-κB binding site restored fluorescence, and the degree of restoration was inversely correlated with the number of nucleotide substitutions within the NF-κB binding site of the competitor. Evaluation of two NF-κB inhibitors, Evans Blue and dehydroxymethylepoxyquinomicin ([−]- DHMEQ), was carried out using p50 and p52 (another form of NF-κB), and IC50 values were obtained. The DSE- FRET technique also detected the differential effect of (−)- DHMEQ on p50 and p52 inhibition. These data indicate that DSE- FRET can be used for high throughput screening of anticancer drugs targeted to DNA-binding proteins. [ABSTRACT FROM AUTHOR]

The top‐seeded solution growth method is a promising technique for growing high‐quality silicon carbide single crystal. Some inherent issues in this growth process, such as morphological instability, polycrystalline growth, and low growth rate, should be clarified. A high temperature difference between the seed and the crucible wall in this system is needed to enhance growth. However, such a high temperature gradient makes the radial growth rate profile non‐uniform due to the effect of Marangoni convection below the seed crystal, which leads to poor crystal quality. In the present work, the effects of crucible size and crucible rotation are numerically investigated to minimize the effect of Marangoni convection. The possibilities of the occurrence of growth‐rate non‐uniformity and undesired impurity incorporation are examined. A smaller crucible (in radius) leads to a more uniform growth rate profile. However, it gives rise to a higher possibility of impurity incorporation. It is also predicted that crucible rotation is ineffective in suppressing the Marangoni flow near the seed edge. This leads to a flow stagnation in the center of the melt, and consequently, it does not enhance the carbon transport below the seed. It also does not reduce the possibility of undesired impurity incorporation. [ABSTRACT FROM AUTHOR]