Monocytes and Monocyte-Derived Antigen-Presenting Cells Have Distinct Gene Signatures in Experimental Model of Multiple Sclerosis

Multiple sclerosis (MS) is a chronic inflammatory disease mediated by a complex interaction between the autoreactive lymphocytes and the effector myeloid cells within the central nervous system (CNS). In a murine model of MS, experimental autoimmune encephalomyelitis (EAE), Ly6Chi monocytes migrate into the CNS and further differentiate into antigen-presenting cells (APCs) during disease progression. Currently, there is no information about gene signatures that can distinguish between monocytes and the monocyte-derived APCs. We developed a surface marker-based strategy to distinguish between these two cell types during the stage of EAE when the clinical symptoms were most severe, and performed transcriptome analysis to compare their gene expression. We report here that the inflammatory CNS environment substantially alters gene expression of monocytes, compared to the monocyte differentiation process within CNS. Monocytes in the CNS express genes that encode proinflammatory cytokines and chemokines, and their expression is mostly maintained when the cells differentiate. Moreover, monocyte-derived APCs express surface markers associated with both dendritic cells and macrophages, and have a significant up-regulation of genes that are critical for antigen presentation. Furthermore, we found that Ccl17, Ccl22, and Ccr7 are expressed in monocyte-derived APCs but not the Ly6Chi monocytes. These findings may shed light on identifying molecular signals that control monocyte differentiation and functions during EAE

Pembuatan Alat Peraga Lemari Pendingin Sebagai Media Pembelajaran Mata Kuliah Teknik Pendingin Di Universitas Negeri Semarang

— Media pembelajaran merupakan segala fisik yang menyajikan pesan serta perangsang peserta didik untuk belajar, sehingga keberadaan media pembelajaran penting untuk membantu dalam proses belajar mengajar. Alat peraga adalah salah satu media pembelajaran, dengan memanfaatkan alat peraga proses pembelajaran akan dapat mempermudah dalam memahami materi yang dipelajari oleh mahasiswa, karena ditampilkan dalam bentuk nyata. Teknik Pendingin adalah salah satu mata kuliah yang ada pada Prodi Pendidikan Teknik Elektro. Permasalahannya apakah alat peraga lemari pendingin layak sebagai media pembelajaran pada mata kuliah Teknik Pendingin jurusan Teknik Elektro Universitas Negeri Semarang. Untuk itu perlu diadakan penelitian untuk mengetahui apakah alat peraga lemari pendingin ini layak untuk dignakan sebagai media pembelajaran. Data dikumpulkan dengan metode angket tertutup maupun terbuka. Alat peraga lemari pendingin ini diujicoba oleh dosen ahli materi teknik pendingin. Metode analisis yang digunakan adalah metode analisis statistik deskriptif. Menurut hasil penelitian dari responden secara keseluruhan, alat peraga lemari pendingin pada mata kuliah Teknik Pendingin ini layak digunakan sebagai media pembelajaran. Dosen ahli materi mengemukakan alat peraga ini layak dijadikan alat peraga setelah adanya revisi alat. Berdasarkan dari hasil penelitian dan pembahasan, dapat disimpulkan bahwa menurut mahasiswa media pembelajaran yang berupa alat peraga lemari pendingin pada mata kuliah Teknik Pendingin diwujudkan dengan menyusun prosedur kerja dengan langkah-langkah sebagai berikut: perencanaan alat peraga, penyediaan alat dan bahan, pembuatan alat peraga, validasi alat peraga, uji coba alat peraga, dan evaluasi. Dari hasil penelitian yang telah dilakukan kepada mahasiswa dengan beberapa aspek, alat peraga lemari pendingin ini termasuk dalam kategori layak, sehingga alat peraga ini dapat digunakan sebagai media pembelajaran. Namun masih terdapat kekurangan pada bahan penutup yang dugunakan seharusnya tidak menggunakan kaca agar tidah mudah pecah, dan jika menggunakan kaca suhu yang ada di dalam lemari pendingin masih dapat terpengaruh oleh suhu udara luar. Kata kunci— Media Pembelajaran, teknik pendingin, universitas negeri semarang, alat peraga, lemari pendingi

Intent in Patent Infringement

In An Intentional Tort Theory of Patents , Professor Vishnubhakat makes two arguments. First, that liability for patent infringement should only be imposed upon defendants who intentionally make, use, or sell, patented inventions. And second, that if patent infringement includes such an intent requirement, it would no longer be a strict liability tort. This response agrees with the first thesis: patent infringement should require intentional conduct of a certain sort. However, the response disagrees with the second thesis: even if patent infringement requires such intent, liability would, in my view, still be “ strict.

c-Myc Is a Universal Amplifier of Expressed Genes in Lymphocytes and Embryonic Stem Cells

SummaryThe c-Myc HLH-bZIP protein has been implicated in physiological or pathological growth, proliferation, apoptosis, metabolism, and differentiation at the cellular, tissue, or organismal levels via regulation of numerous target genes. No principle yet unifies Myc action due partly to an incomplete inventory and functional accounting of Myc’s targets. To observe Myc target expression and function in a system where Myc is temporally and physiologically regulated, the transcriptomes and the genome-wide distributions of Myc, RNA polymerase II, and chromatin modifications were compared during lymphocyte activation and in ES cells as well. A remarkably simple rule emerged from this quantitative analysis: Myc is not an on-off specifier of gene activity, but is a nonlinear amplifier of expression, acting universally at active genes, except for immediate early genes that are strongly induced before Myc. This rule of Myc action explains the vast majority of Myc biology observed in literature

Analysis of single nuclear chromatin accessibility reveals unique myeloid populations in human pancreatic ductal adenocarcinoma

Background: A better understanding of the pancreatic ductal adenocarcinoma (PDAC) immune microenvironment is critical to developing new treatments and improving outcomes. Myeloid cells are of particular importance for PDAC progression; however, the presence of heterogenous subsets with different ontogeny and impact, along with some fluidity between them, (infiltrating monocytes vs. tissue‐resident macrophages; M1 vs. M2) makes characterisation of myeloid populations challenging. Recent advances in single cell sequencing technology provide tools for characterisation of immune cell infiltrates, and open chromatin provides source and function data for myeloid cells to assist in more comprehensive characterisation. Thus, we explore single nuclear assay for transposase accessible chromatin (ATAC) sequencing (snATAC‐Seq), a method to analyse open gene promoters and transcription factor binding, as an important means for discerning the myeloid composition in human PDAC tumours. Methods: Frozen pancreatic tissues (benign or PDAC) were prepared for snATAC‐Seq using 10× Chromium technology. Signac was used for preliminary analysis, clustering and differentially accessible chromatin region identification. The genes annotated in promoter regions were used for Gene Ontology (GO) enrichment and cell type annotation. Gene signatures were used for survival analysis with The Cancer Genome Atlas (TCGA)‐pancreatic adenocarcinoma (PAAD) dataset. Results: Myeloid cell transcription factor activities were higher in tumour than benign pancreatic samples, enabling us to further stratify tumour myeloid populations. Subcluster analysis revealed eight distinct myeloid populations. GO enrichment demonstrated unique functions for myeloid populations, including interleukin‐1b signalling (recruited monocytes) and intracellular protein transport (dendritic cells). The identified gene signature for dendritic cells influenced survival (hazard ratio = .63, p = .03) in the TCGA‐PAAD dataset, which was unique to PDAC. Conclusions: These data suggest snATAC‐Seq as a method for analysis of frozen human pancreatic tissues to distinguish myeloid populations. An improved understanding of myeloid cell heterogeneity and function is important for developing new treatment targets in PDAC

Unleashing the Power of ChatGPT in Finance Research: Opportunities and Challenges

Natural language processing (NLP) technologies, such as ChatGPT, are revolutionizing various fields, including finance research. This article explores the multifaceted potential of ChatGPT as a transformative tool for finance researchers, highlighting the benefits, challenges, and novel insights it can offer to facilitate the research. We demonstrate applications in coding support, theoretical derivation, research idea assistance, and professional editing. A comparison of ChatGPT-3.5, ChatGPT-4, and Microsoft Bing reveals unique features and applicability. By discussing pitfalls and ethical concerns, we encourage responsible AI adoption and a comprehensive understanding of advanced NLP’s impact on finance research and practice

Phosphorylation of H4 Ser 47 promotes HIRA-mediated nucleosome assembly

Histone H3 variant H3.3, while differing from canonical H3 (H3.1) by only five amino acids, is assembled into nucleosomes, along with histone H4, at genic regions by the histone chaperone HIRA, whereas H3.1 is assembled into nucleosomes in a CAF-1-dependent reaction. Here, we show that phosphorylation of histone H4 Ser 47 (H4S47ph), catalyzed by the PAK2 kinase, promotes nucleosome assembly of H3.3–H4 and inhibits nucleosome assembly of H3.1–H4 by increasing the binding affinity of HIRA to H3.3–H4 and reducing association of CAF-1 with H3.1–H4. These results reveal a mechanism whereby H4S47ph distinctly regulates nucleosome assembly of H3.1 and H3.3

Emergence of Resistance to MTI-101 Selects for a MET Genotype and Phenotype in EGFR Driven PC-9 and PTEN Deleted H446 Lung Cancer Cell Lines

MTI-101 is a first-in-class cyclic peptide that kills cells via calcium overload in a caspase-independent manner. Understanding biomarkers of response is critical for positioning a novel therapeutic toward clinical development. Isogenic MTI-101-acquired drug-resistant lung cancer cell line systems (PC-9 and H446) coupled with differential RNA-SEQ analysis indicated that downregulated genes were enriched in the hallmark gene set for epithelial-to-mesenchymal transition (EMT) in both MTI-101-acquired resistant cell lines. The RNA-SEQ results were consistent with changes in the phenotype, including a decreased invasion in Matrigel and expression changes in EMT markers (E-cadherin, vimentin and Twist) at the protein level. Furthermore, in the EGFR-driven PC-9 cell line, selection for resistance towards MTI-101 resulted in collateral sensitivity toward EGFR inhibitors. MTI-101 treatment showed synergistic activity with the standard of care agents erlotinib, osimertinib and cisplatin when used in combination in PC-9 and H446 cells, respectively. Finally, in vivo data indicate that MTI-101 treatment selects for increased E-cadherin and decreased vimentin in H446, along with a decreased incident of bone metastasis in the PC-9 in vivo model. Together, these data indicate that chronic MTI-101 treatment can lead to a change in cell state that could potentially be leveraged therapeutically to reduce metastatic disease

Author Correction: Mapping histone modifications in low cell number and single cells using antibody-guided chromatin tagmentation (ACT-seq)

An amendment to this paper has been published and can be accessed via a link at the top of the paper

Transient Receptor Potential C 1/4/5 Is a Determinant of MTI-101 Induced Calcium Influx and Cell Death in Multiple Myeloma

Multiple myeloma (MM) is a currently incurable hematologic cancer. Patients that initially respond to therapeutic intervention eventually relapse with drug resistant disease. Thus, novel treatment strategies are critically needed to improve patient outcomes. Our group has developed a novel cyclic peptide referred to as MTI-101 for the treatment of MM. We previously reported that acquired resistance to HYD-1, the linear form of MTI-101, correlated with the repression of genes involved in store operated Ca2+ entry (SOCE): PLCβ, SERCA, ITPR3, and TRPC1 expression. In this study, we sought to determine the role of TRPC1 heteromers in mediating MTI-101 induced cationic flux. Our data indicate that, consistent with the activation of TRPC heteromers, MTI-101 treatment induced Ca2+ and Na+ influx. However, replacing extracellular Na+ with NMDG did not reduce MTI-101-induced cell death. In contrast, decreasing extracellular Ca2+ reduced both MTI-101-induced Ca2+ influx as well as cell death. The causative role of TRPC heteromers was established by suppressing STIM1, TRPC1, TRPC4, or TRPC5 function both pharmacologically and by siRNA, resulting in a reduction in MTI-101-induced Ca2+ influx. Mechanistically, MTI-101 treatment induces trafficking of TRPC1 to the membrane and co-immunoprecipitation studies indicate that MTI-101 treatment induces a TRPC1-STIM1 complex. Moreover, treatment with calpeptin inhibited MTI-101-induced Ca2+ influx and cell death, indicating a role of calpain in the mechanism of MTI-101-induced cytotoxicity. Finally, components of the SOCE pathway were found to be poor prognostic indicators among MM patients, suggesting that this pathway is attractive for the treatment of MM