NAMPT-Driven M2 Polarization of Tumor-Associated Macrophages Leads to an Immunosuppressive Microenvironment in Colorectal Cancer.

Nicotinamide phosphoribosyltransferase (NAMPT) is a metabolic enzyme with key roles in inflammation. Previous studies have examined the consequences of its upregulated expression in cancer cells themselves, but studies are limited with respect to its role in the other cells within the tumor microenvironment (TME) during colorectal cancer (CRC) progression. Using single-cell RNA sequencing (scRNA-seq) data, it is founded that NAMPT is highly expressed in SPP1+ tumor-associated macrophages (TAMs), a unique subset of TAMs associated with immunosuppressive activity. A NAMPThigh gene signature in SPP1+ TAMs correlated with worse prognostic outcomes in CRC patients. The effect of Nampt deletion in the myeloid compartment of mice during CRC development is explored. NAMPT deficiency in macrophages resulted in HIF-1α destabilization, leading to reduction in M2-like TAM polarization. NAMPT deficiency caused significant decreases in the efferocytosis activity of macrophages, which enhanced STING signaling and the induction of type I IFN-response genes. Expression of these genes contributed to anti-tumoral immunity via potentiation of cytotoxic T cell activity in the TME. Overall, these findings suggest that NAMPT-initiated TAM-specific genes can be useful in predicting poor CRC patient outcomes; strategies aimed at targeting NAMPT may provide a promising therapeutic approach for building an immunostimulatory TME in CRC progression.

LCK-Mediated RIPK3 Activation Controls Double-Positive Thymocyte Proliferation and Restrains Thymic Lymphoma by Regulating the PP2A-ERK Axis.

Receptor-interacting protein kinase 3 (RIPK3) is the primary regulator of necroptotic cell death. RIPK3 expression is often silenced in various cancer cells, which suggests that it may have tumor suppressor properties. However, the exact mechanism by which RIPK3 negatively regulates cancer development and progression remains unclear. This report indicates that RIPK3 acts as a potent regulator of the homeostatic proliferation of CD4+ CD8+ double-positive (DP) thymocytes. Abnormal proliferation of RIPK3-deficient DP thymocytes occurs independently of the well-known role for RIPK3 in necroptosis (upstream of MLKL activation), and is associated with an incidental thymic mass, likely thymic hyperplasia. In addition, Ripk3-null mice develop increased thymic tumor formation accompanied by reduced host survival in the context of an N-ethyl-N-nitrosourea (ENU)-induced tumor model. Moreover, RIPK3 deficiency in p53-null mice promotes thymic lymphoma development via upregulated extracellular signal-regulated kinase (ERK) signaling, which correlates with markedly reduced survival rates. Mechanistically, lymphocyte-specific protein tyrosine kinase (LCK) activates RIPK3, which in turn leads to increases in the phosphatase activity of protein phosphatase 2 (PP2A), thereby suppressing hyper-activation of ERK in DP thymocytes. Overall, these findings suggest that a RIPK3-PP2A-ERK signaling axis regulates DP thymocyte homeostasis and may provide a potential therapeutic target to improve thymic lymphoma therapies.

Highly selective and sensitive detection of neurotransmitters using receptor-modified single-walled carbon nanotube sensors.

We present receptor-modified carbon nanotube sensors for the highly selective and sensitive detection of acetylcholine (ACh), one kind of neurotransmitter. Here, we successfully expressed the M1 muscarinic acetylcholine receptor (M1 mAChR), a family of G protein-coupled receptors (GPCRs), in E. coli and coated single-walled carbon nanotube (swCNT)-field effect transistors (FETs) with lipid membrane including the receptor, enabling highly selective and sensitive ACh detection. Using this sensor, we could detect ACh at 100 pM concentration. Moreover, we showed that this sensor could selectively detect ACh among other neurotransmitters. This is the first demonstration of the real-time detection of ACh using specific binding between ACh and M1 mAChR, and it may lead to breakthroughs for various applications such as disease diagnosis and drug screening.

Parent-reported ISAAC written questionnaire may underestimate the prevalence of asthma in children aged 10-12 years.

OBJECTIVES: The aim of the present study was to compare the validity of the International Study of Asthma and Allergies in Childhood (ISAAC) written (WQ) and audiovisual questionnaires (AVQ 3.0) in two age-groups (10-12 and 13-15 years, respectively). METHODS: The 13-15 year olds performed the self-completed the WQ and AVQ on the same day. The 10-12 year olds performed the self-completed the AVQ and the parent-completed WQ was completed by their parents. The methacholine challenge test was conducted in 10-12 year olds from one elementary school. RESULTS: In 10-12 year olds, the AVQ detected a generally higher prevalence of asthma symptoms than WQ. In 13-15 year olds, this was reversed. In 10-12 year olds, poor agreement was found between the parent-completed WQ and the self-reported AVQ. In 13-15 year olds, moderate agreement was found between the self-reported WQ and AVQ. Low sensitivity was found, in predicting bronchial hyper-responsiveness (BHR) for all questions of both WQ and AVQ in 10-12 year olds. However, the AVQ had slightly higher sensitivity than WQ, with the exception of wheeze ever, although it was not statistically significant. CONCLUSION: The ISAAC AVQ may be another effective instrument for assessing the prevalence of asthma symptoms in children aged 10-12 years, whereas the parent-reported-WQ may underestimate the prevalence of asthma symptoms in this age-group.

Nanoneedle transistor-based sensors for the selective detection of intracellular calcium ions.

We developed a nanoneedle transistor-based sensor (NTS) for the selective detection of calcium ions inside a living cell. In this work, a single-walled carbon nanotube-based field effect transistor (swCNT-FET) was first fabricated at the end of a glass nanopipette and functionalized with Fluo-4-AM probe dye. The selective binding of calcium ions onto the dye molecules altered the charge state of the dye molecules, resulting in the change of the source-drain current of the swCNT-FET as well as the fluorescence intensity from the dye. We demonstrated the electrical and fluorescence detection of the concentration change of intracellular calcium ions inside a HeLa cell using the NTS.

Uniform patterning of sub-50-nm-scale Au nanostructures on insulating solid substrate via dip-pen nanolithography.

We report a direct deposition strategy for sub-50-nm-scale uniform Au patterns on virtually any general insulating substrate via dip-pen nanolithography (DPN). In that process, HAuCl(4) molecules were deposited onto bare insulating substrates via a molecular diffusion process, in the absence of electrochemical reactions. Subsequently, the generated HAuCl(4) molecular patterns were decomposed to leave Au-only patterns using a thermal annealing process. Uniform Au patterns with a mean diameter of 47.9 +/- 3.1 nm were achieved after the annealing process. The strategy allowed us to generate Au patterns on virtually any general insulating substrate (e.g., SiO(2), Al(2)O(3), polyimide, etc) without the need for surface functionalization or additional electrode structures. This versatile and reliable patterning method is expected to be useful in the future development of various novel industrial applications (e.g., mask or nanocircuit repair, nanosensors, etc.).

Massive assembly of ZnO nanowire-based integrated devices.

Although a directed assembly strategy has been utilized for the massive assembly of various nanowires and nanotubes (NWs/NTs), its application has usually been limited to rather small-diameter NWs/NTs prepared in solution. We report two complementary methods for the massive assembly of large-size ZnO nanowires (NWs). In the solution-phase method, ZnO NWs were assembled and aligned selectively onto negatively charged surface patterns in solution. In addition, the substrate bias voltage and capillary forces can be used to further enhance the adsorption rate and degree of alignment of ZnO NWs, respectively. In the direct-transfer method, a NW film grown on a solid substrate was placed in close proximity to a molecule-patterned substrate, and ultrasonic vibration was applied so that the NWs were directly transferred and aligned onto the patterned substrate. The solution-phase and direct-transfer methods are complementary to each other and suitable for the assembly of NWs prepared in solution and on solid substrates, respectively.

Selective adsorption and alignment behaviors of double- and multiwalled carbon nanotubes on bare Au and SiO2 surfaces.

We present the study of selective adsorption and alignment behaviors of double- and multiwalled carbon nanotubes (dwCNTs and mwCNTs) on self-assembled monolayer (SAM) patterns, bare Au, and SiO2 surfaces. dwCNTs and mwCNTs exhibited stronger affinity to polar SAMs, bare Au, and SiO2 surfaces than to nonpolar SAM surfaces. Furthermore, we found the adsorption probability of smaller carbon nanotubes (CNTs) was higher than that of larger CNTs. As proof of concept, we successfully assembled and aligned dwCNTs and mwCNTs on Au and SiO2 substrates without relying on external forces and demonstrated wafer-scale fabrication of back-gate transistors based on dwCNTs with a high yield.

Phase of molecular ink in nanoscale direct deposition processes.

We report the first observation of a phase transition in a nanoscale direct deposition process. This transition involves the melting of molecular ink layers in dip-pen nanolithography, and it is observed by measuring the temperature dependence of the growth rate of the deposited pattern. The results are interpreted using a diffusion equation approach in conjunction with a "double-molecular-layer" model of the adsorbed molecules on the atomic force microscope tip. The theory provides a qualitative explanation for the dependence of the pattern growth rate on solvent and adsorbed water as well as on temperature.