bluuebunny/crossref_metadata_2025 huggingface dataset:
Sistem Pendukung Keputusan Kelayakan Klaim Asuransi Menggunakan Metode AHP
Authors: Islam Abdullah Izzul , Asep Jamaludin  , Nono Heryana
Published: February 2021

Pertumbuhan asuransi di Indonesia belakangan ini mengalami peningkatan yang cukup baik hal ini dibuktikan pada statistik pelaku IKNB (Industri Keuangan Non Bank)  yang diterbitkan oleh OJK (Otoritas Jasa Keuangan) pada Februari 2020. Pada desember 2018 jumlah pelaku IKNB berjumlah 1260 lalu mengalami kenaikan setiap bulannya hingga pada desember 2019 pelaku IKNB berjumlah 1330. Dari data tersebut pelaku IKNB mengalami peningkatan yang cukup baik, dimana asuransi termasuk dalam industri tersebut. Dengan banyaknya pelaku IKNB tersebut PT. SICO Andalan Utama yang merupakan salah satu perusahaan yang bergerak dalam industry tersebut merasakan hal peningkatan hal tersebut. Namun dengan peningkatan tersebut dibutuhkan pengelolaan asuransi terkhusus pengelolaan klaim karena butuh kehati-hatian dalam memutuskan keputusan kelayakan klaim. Maka untuk mengatasi permasalahan tersebut hal inilah yang mendasari penulis untuk mengusulkan pengembangan Sistem Pendukung Keputusan Kelayakan Klaim Asuransi yang nantinya diahrapkan dapat menghindari atau minimal mengurangi ketidak akuratan dalam  menentukan kelayakan klaim. Sistem Pendukung Keputusan Kelayakan Klaim Asuransi ini dikembangkan dengan metodologi pengembangan sistem  Rapid Application Development (RAD) sebagai alur dari pengembangan system. Sedangkan untuk pemodelan SPK nya menggunakan model  Analytical Hierarchy Process (AHP). Dan  juga menggunakan beberapa tools Unified Modelling Language (UML) , Business Process Modelling Nation (BPMN), dan Entity Relationship Diagram (ERD) untuk analisis maupun perancangannya. Dalam pengimplementasian system penulis menggunakan PHP sebagai Bahasa pemrograman dan MySQL sebagai Database. Metode pengujian menggunakan metode pendekatan White Box Testing, Black Box Testing Serta Usability Testing. Dengan diterapkan system ini diharapkan dapat memberikan keputusan kelayakan klaim yang tepat.
Sistem Pakar Perencanaan Penjadwalan Bangunan Rumah Minimalis Berdasarkan Tipe dengan Metode Algoritma Genetika
Authors: Aspudji Muhammad Aspandi , Budi Harijanto  , Rizky Ardiansyah
Published: February 2021

Pelaksanaan pembangunan rumah sering kali terjadi keterlambatan dikarenakan penjadwalan yang kurang optimal. Menjawab permasalahan tersebut, maka solusi yang diperlukan adalah sebuah aplikasi penjadwalan pembangunan rumah berbasis web yang sistematis dan terintegrasi dengan melibatkan beberapa komponen penting, sehingga meminimalisir keterlambatan penjadwalan, merinci kebutuhan bahan sesuai dengan persiapan dana yang ada. Dengan menerapkan metode Algoritma Genetika untuk melakukan optimasi biaya dan penjadwalan pembangunan. Dilakukan percobaan nilai kombinasi parameter pada rumah tipe 36 dan didapatkan nilai kombinasi terbaik dengan nilai Kromosom: 100, Jumlah generasi: 80, Probabilitas Crossover: 0,7 dan Probabilitas Mutasi: 0,05 didapatkan nilai pengoptimalan durasi Penjadwalan Pembangunan 84 hari, Optimasi Biaya Rp. 102.627.995 dan nilai fitness: 0,0937 dengan tingkat akurasi sebesar 91,6%.
Identifikasi "Acne Vulgaris" Berdasarkan Fitur Warna Dan Tekstur Menggunakan Klasifikasi JST Backpropagation
Authors: Rosiani Ulla Delfana , Batubulan Kadek Suarjuna , Malia Elisiana
Published: February 2021

Acne Vulgaris adalah salah satu penyakit kulit kronis umum yang berkaitan dengan penyumbatan dan atau peradangan pada folikel rambut dan kelenjar minyak yang menyertainya. Acne vulgaris dibagi menjadi beberapa macam jenis, diantaranya ada whitehead, blackhead, papule, dan pustule. Beberapa jenis acne vulgaris memiliki warna dan tekstur yang berbeda. Hasil dari ekstraksi warna dan tekstur dapat dilakukan klasifikasi terhadap jenis acne vulgaris. Dengan berkembangnya teknologi bidang informatics engineering proses identifikasi jenis acne vulgaris dapat dilakukan menggunakan pengolahan citra digital. Hasil dari ekstraksi fitur warna dan fitur tekstur kemudian dianalisis dan diklasifikasikan sehingga dapat diketahui jenis acne vulgaris pada citra tersebut. HSV merupakan salah satu ruang warna yang memperlihatkan warna seperti yang dilihat oleh manusia pada umumnya. GLCM merupakan metode yang mampu menyediakan informasi penting dan melakukan analisis mengenai tekstur citra. Empat ciri statistik GLCM yaitu contrast, correlations, homogenity, dan energy dengan sudut 0°, 45°, 90°, dan 135° akan memberikan nilai untuk membedakan tekstur acne vulgaris pada setiap jenisnya. Backpropagation digunakan karena metode tersebut mampu menyelesaikan pengklasifikasian dengan problem nonlinear. Terdapat tiga tahap backpropagation dalam pengolahan data yaitu tahap feedfordward, backpropagation, dan penyesuaian bobot, sedangkan untuk klasifikasi hanya menggunakan tahap feedfordward. Dari hasil penelitian mengidentifikasi acne vulgaris menggunakan ruang warna HSV, metode GLCM dan klasifikasi JST Backpropagation dapat menghasilkan nilai akurasi sebesar 65%. Akurasi tersebut didapatkan dari parameter learning rate 0.0002 dan epoch sebanyak 200.
Ekstraksi Click Stream Data Web E-Commerce Menggunakan Web Usage Mining
Authors: Kartina Diah Kusuma Wardani
Published: February 2021

E-Commerce berkembang pesat dalam world wide web hingga menghasilkan berbagai jenis data yang dapat dianalisa lebih lanjut untuk berbagai keperluan seperti personifikasi web, profiling customer, dan sebagainya. Salah satu jenis data yang dihasilkan e-Commerce adalah click stream data web yang merekam aktivitas visitor web dalam bentuk log data selama berinteraksi pada laman web. Penelitian ini mengekstraksi click stream data web e-commerce untuk mendapatkan pola interaksi konsumen terhadap halaman web selama mengunjungi web e-commerce. Berdasarkan jenis data yang diekstrak maka web usage mining digunakan untuk ekstraksi pola dari click stream data yang berbentuk log data. Teknik mining yang dianalisa terhadap log data e-commerce pada penelitian ini terdiri dari frequent itemset, asociation rules, dan frequence sequence mining. Frequent itemset menghasilkan halaman web yang paling sering diakses oleh visitor. Association rules menghasilkan pola kemungkinan halaman web yang akan diakses visitor jika visitor mengakses halaman-halamn tertentu. Frequence sequence mining mendapatkan pola urutan halaman web yang paling sering diakses oleh visitor web e-commerce saat berinteraksi pada laman web. Pola urutan halaman yang diakses visitor menunjukkan urutan kebiasaan visitor mengunjungi e-commerce. Sedangkan teknik mining yang diimplementasikan untuk menghasilkan pola akses visitor pada penelitian ini adalah Frequence sequence mining. Hasil ekstraksi dari penelitian ini menunjukkan ada enam halaman web yang paling sering diakses oleh konsumen dengan berbagai pola urutan aksesnya.
Pengembangan Sistem Analisis Sentimen Berbasis Java Pada Data Twitter Terhadap Omnibus Law Menggunakan Algoritma Naïve Bayes dan K-Nearst Neighbor (K-NN)
Authors: Hamdana Elok Nur , Alfahmi Muhammad Balya Iqbal
Published: February 2021

Undang-undang Cipta Kerja Omnibus law telah di sah kan, pengesahan tersebut mendapat penolakan dari berbagai elemen masyarakat. Hal itu disebabkan Omnibus Law UU Cipta Kerja, dinilai akan membawa dampak buruk bagi tenaga kerja atau buruh. Oleh karena itu peneliti melakukan analisis sentimen dalam bidang data mining terhadap UU Cipta Kerja Omnibus law pada media sosial Twitter. Penulis menggunakan metode Naïve Bayes dan KNN sebagai algoritma yang diterapkan dalam system berbasis java untuk memperbandingkan tingkat akurasi dari kedua metode tersebut. Peneliti menggunakan metode web scrapping untuk proses penggambilan data dari twitter secara real time. Hasil penelitian menunjukan bahwa analisis sentimen terhadap data Twitter terhadap UU Cipta Kerja Ombibus law dengan menggunakan metode Naïve Bayes mencapai tingkat akurasi 75% dengan class precision untuk pred. positive adalah 57%, pred negative adalah 72%, dan pred. neutral adalah 54%. Lalu pada metode KNN tingkat akurasi mencapai 88%. Dimana class precision untuk pred. positive adalah 94%, pred negative adalah 61%, dan pred. neutral adalah 58%.
FBS-Derived Exosomes as a Natural Nano-Scale Carrier for Icariin Promote Osteoblast Proliferation
Authors: Dong Ming , Wu Saixuan , Xu Huijun , Yu Xinxin , Wang Lina , Bai Hua , Niu Weidong
Published: February 2021

Icariin is a class IV drug of low solubility, permeability, and poor bioavailability. Synthetic nanomaterials have developed rapidly. However, some literatures point out that synthetic nanomaterials such as liposomes, aptamers, metal nanoparticles, and nanogels have high toxicity and are affected by the reticuloendothelial system or mononuclear phagocyte system. It is known that exosomes could be used as an ideal clinical drug delivery vehicle to avoid the above-mentioned problems to a certain extent. Studies have shown that drugs can be loaded into exosomes by passive and active loading. We used Fetal bovine serum (FBS) exosomes to carry Icariin for the first time in this experiment, FBS exosomes-Icariin (FBS EXO-ICA) more effectively promoted the proliferation of osteoblasts and bone regeneration than Icariin alone. FBS EXO-ICA could become a new nano scale drug formulation for treating diseases associated with bone loss.
Minimize the Xylitol Production in Saccharomyces cerevisiae by Balancing the Xylose Redox Metabolic Pathway
Authors: Zhu Yixuan , Zhang Jingtao , Zhu Lang , Jia Zefang , Li Qi , Xiao Wei , Cao Limin
Published: February 2021

Xylose is the second most abundant sugar in lignocellulose, but it cannot be used as carbon source by budding yeast Saccharomyces cerevisiae. Rational promoter elements engineering approaches were taken for efficient xylose fermentation in budding yeast. Among promoters surveyed, HXT7 exhibited the best performance. The HXT7 promoter is suppressed in the presence of glucose and derepressed by xylose, making it a promising candidate to drive xylose metabolism. However, simple ectopic expression of both key xylose metabolic genes XYL1 and XYL2 by the HXT7 promoter resulted in massive accumulation of the xylose metabolic byproduct xylitol. Through the HXT7-driven expression of a reported redox variant, XYL1-K270R, along with optimized expression of XYL2 and the downstream pentose phosphate pathway genes, a balanced xylose metabolism toward ethanol formation was achieved. Fermented in a culture medium containing 50 g/L xylose as the sole carbon source, xylose is nearly consumed, with less than 3 g/L xylitol, and more than 16 g/L ethanol production. Hence, the combination of an inducible promoter and redox balance of the xylose utilization pathway is an attractive approach to optimizing fuel production from lignocellulose.
Temperature-Dependent Activity of Motor Proteins: Energetics and Their Implications for Collective Behavior
Authors: Yadav Saumya , Kunwar Ambarish
Published: February 2021

Molecular motor proteins are an extremely important component of the cellular transport system that harness chemical energy derived from ATP hydrolysis to carry out directed mechanical motion inside the cells. Transport properties of these motors such as processivity, velocity, and their load dependence have been well established through single-molecule experiments. Temperature dependent biophysical properties of molecular motors are now being probed using single-molecule experiments. Additionally, the temperature dependent biochemical properties of motors (ATPase activity) are probed to understand the underlying mechanisms and their possible implications on the enzymatic activity of motor proteins. These experiments in turn have revealed their activation energies and how they compare with the thermal energy available from the surrounding medium. In this review, we summarize such temperature dependent biophysical and biochemical properties of linear and rotary motor proteins and their implications for collective function during intracellular transport and cellular movement, respectively.
Integrated Protein–Protein Interaction and Weighted Gene Co-expression Network Analysis Uncover Three Key Genes in Hepatoblastoma
Authors: Tian Linlin , Chen Tong , Lu Jiaju , Yan Jianguo , Zhang Yuting , Qin Peifang , Ding Sentai , Zhou Yali
Published: February 2021

Hepatoblastoma (HB) is the most common liver tumor in the pediatric population, with typically poor outcomes for advanced-stage or chemotherapy-refractory HB patients. The objective of this study was to identify genes involved in HB pathogenesis via microarray analysis and subsequent experimental validation. We identified 856 differentially expressed genes (DEGs) between HB and normal liver tissue based on two publicly available microarray datasets (GSE131329 and GSE75271) after data merging and batch effect correction. Protein–protein interaction (PPI) analysis and weighted gene co-expression network analysis (WGCNA) were conducted to explore HB-related critical modules and hub genes. Subsequently, Gene Ontology (GO) analysis was used to reveal critical biological functions in the initiation and progression of HB. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that genes involved in cell cycle phase transition and the PI3K/AKT signaling were associated with HB. The intersection of hub genes identified by both PPI and WGCNA analyses revealed five potential candidate genes. Based on receiver operating characteristic (ROC) curve analysis and reports in the literature, we selected CCNA2, CDK1, and CDC20 as key genes of interest to validate experimentally. CCNA2, CDK1, or CDC20 small interfering RNA (siRNA) knockdown inhibited aggressive biological properties of both HepG2 and HuH-6 cell linesin vitro. In conclusion, we identified CCNA2, CDK1, and CDC20 as new potential therapeutic biomarkers for HB, providing novel insights into important and viable targets in future HB treatment.
Engineering Human Cardiac Muscle Patch Constructs for Prevention of Post-infarction LV Remodeling
Authors: Wang Lu , Serpooshan Vahid , Zhang Jianyi
Published: February 2021

Tissue engineering combines principles of engineering and biology to generate living tissue equivalents for drug testing, disease modeling, and regenerative medicine. As techniques for reprogramming human somatic cells into induced pluripotent stem cells (iPSCs) and subsequently differentiating them into cardiomyocytes and other cardiac cells have become increasingly efficient, progress toward the development of engineered human cardiac muscle patch (hCMP) and heart tissue analogs has accelerated. A few pilot clinical studies in patients with post-infarction LV remodeling have been already approved. Conventional methods for hCMP fabrication include suspending cells within scaffolds, consisting of biocompatible materials, or growing two-dimensional sheets that can be stacked to form multilayered constructs. More recently, advanced technologies, such as micropatterning and three-dimensional bioprinting, have enabled fabrication of hCMP architectures at unprecedented spatiotemporal resolution. However, the studies working on various hCMP-based strategies for in vivo tissue repair face several major obstacles, including the inadequate scalability for clinical applications, poor integration and engraftment rate, and the lack of functional vasculature. Here, we review many of the recent advancements and key concerns in cardiac tissue engineering, focusing primarily on the production of hCMPs at clinical/industrial scales that are suitable for administration to patients with myocardial disease. The wide variety of cardiac cell types and sources that are applicable to hCMP biomanufacturing are elaborated. Finally, some of the key challenges remaining in the field and potential future directions to address these obstacles are discussed.
Encapsulating Sulfur Into Nickel Decorated Hollow Carbon Fibers for High-Performance Lithium-Sulfur Batteries
Authors: Yu Dongdong , Tang Zhihong , He Haiyong
Published: February 2021

Due to the high specific energy density, lithium-sulfur batteries (LSBs) have great potential in energy storage devices for electric vehicle and electronic equipment. However, poor conductivity of sulfur, large volume expansion, and lithium polysulfide dissolution limit LSBs application and promotion. In this work, graphitic hollow carbon fibers (HCF) were fabricated as a matrix to encapsulate sulfur. And nickel particles were introduced into fibers (Ni@HCF) to improve the cycle stability of sulfur cathode. On one hand, hollow structures can encapsulate sulfur and limit lithium polysulfides dissolution, and the graphitic carbon walls can provide a fast electron transport channel. On the other hand, nickel particles can accelerate the conversion of lithium polysulfides. The study results show that the initial discharge specific capacity of Ni@HCF/S cathodes reaches 1,252 mAh g−1 at the current density of 0.1C. And the capacity can be maintained at 558 mAh g−1 after 200 cycles at the current density of 1C.
Socio-Ecological Networks: A Lens That Focuses Beyond Physics
Authors: Ulanowicz Robert E.
Published: February 2021

Contemporary systems ecology has long been occupied with mechanical explanations of behavior; however, the physical theory that undergirds such explanations has certain limits. It’s not that the physical force laws are ever violated, but with heterogeneous, irreversible relationships subject to aleatoric influences, physical laws can only constrain, but not determine, outcomes. Such complex systems are better treated in the framework of quantified networks of interrelations. The application of simple information theory to networks reveals that ecosystems cannot achieve maximal efficiency without growing vulnerable to novel disturbances. A degree of inefficient redundancies is always necessary to sustain ecological and social functioning; and, if they are to function reliably, such non-optimal features become requisite in the design of infrastructures like power grids, water distribution networks, traffic patterns, and supply chains. In particular, the postulate that economic market efficiency should never be compromised must be re-examined if society is to remain sustainable. Furthermore, the capability of networks to represent distributed causalities allows one to rationalize behaviors like endogenous selection, centripetal acquisition of resources, and the precedence of indirect mutualism over competition in living systems–all phenomena that challenge conventional evolutionary dogma.
The Major Intrinsic Protein Family and Their Function Under Salt-Stress in Peanut
Authors: Han Yan , Li Rongchong , Liu Yiyang , Fan Shoujin , Wan Shubo , Zhang Xuejie , Li Guowei
Published: February 2021

Peanut (Arachis hypogaea) is an important oil crop cultivated across the world. Abiotic stresses are the major constraint factors that defect its yield, especially in the rainfed peanut cultivation areas. Aquaporins are proteins that form a large family of more than 30 members in higher plants and play key roles in plant water balance under abiotic stress conditions. To comprehensively understand the functions of aquaporins in peanut, we identified their family genome-wide and characterized the phylogenetics, gene structure, and the conserved motif of the selective filter. In total, 64 aquaporin isoforms were identified, the NIPs were firstly categorized into NIP1s and NIP2s groups based on the phylogenetic analysis and the selective filter structure classification system. Further, we analyzed the gene expression pattern under the salt-stress conditions and found that a TIP3 member is strongly induced by salt stress, which in turn contributed to improved seed germination under salt stress when expressed in Arabidopsis. Our study thus provides comprehensive profiles on the MIP superfamily and their expression and function under salt-stress conditions. We believe that our findings will facilitate the better understanding of the roles of aquaporins in peanuts under salt salt-stress conditions.
A Negative Feedback Model to Explain Regulation of SARS-CoV-2 Replication and Transcription
Authors: Li Xin , Cheng Zhi , Wang Fang , Chang Jia , Zhao Qiang , Zhou Hao , Liu Chang , Ruan Jishou , Duan Guangyou , Gao Shan
Published: February 2021

Background

Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although a preliminary understanding of the replication and transcription of SARS-CoV-2 has recently emerged, their regulation remains unknown.

Results

By comprehensive analysis of genome sequence and protein structure data, we propose a negative feedback model to explain the regulation of CoV replication and transcription, providing a molecular basis of the “leader-to-body fusion” model. The key step leading to the proposal of our model was that the transcription regulatory sequence (TRS) motifs were identified as the cleavage sites of nsp15, a nidoviral RNA uridylate-specific endoribonuclease (NendoU). According to this model, nsp15 regulates the synthesis of subgenomic RNAs (sgRNAs), and genomic RNAs (gRNAs) by cleaving TRSs. The expression level of nsp15 controls the relative proportions of sgRNAs and gRNAs, which in turn change the expression level of nsp15 to reach equilibrium between the CoV replication and transcription.

Conclusion

The replication and transcription of CoVs are regulated by a negative feedback mechanism that influences the persistence of CoVs in hosts. Our findings enrich fundamental knowledge in the field of gene expression and its regulation, and provide new clues for future studies. One important clue is that nsp15 may be an important and ideal target for the development of drugs (e.g., uridine derivatives) against CoVs.
Sunitinib Exerts In Vitro Immunomodulatory Activity on Sarcomas via Dendritic Cells and Synergizes With PD-1 Blockade
Authors: Ocadlikova Darina , Lecciso Mariangela , Broto Javier Martin , Scotlandi Katia , Cavo Michele , Curti Antonio , Palmerini Emanuela
Published: February 2021

Background

High-grade sarcomas are a heterogeneous group of aggressive tumors arising in bone and soft tissues. After relapse, treatment options are limited. The multi-targeted receptor tyrosine kinase inhibitors (TKIs) sunitinib and inhibitor of PD-1 (anti-PD-1) nivolumab have shown antitumor activity in selected subtypes. In this study, we examine the role of TKIs and PD-1 based therapy in in vitro cocultures of sarcoma.

Methods

The human osteosarcoma (SaOS-2) and synovial sarcoma (SYO-1) cell lines were treated with sunitinib. After cell death and proliferation assessment, expression of PD-L1 was analyzed by flow cytometry. Sunitinib-treated sarcoma cells were cocultured with dendritic cells (DCs), and the phenotype of mature DCs was determined by flow cytometry. Mature DCs were cultured with autologous T cells. PD-1 expression on T cells, their proliferation, T regulatory cell (Tregs) induction and IFN-γ production, before and after nivolumab exposure, were analyzed.

Results

Along with its anti-proliferative and direct pro-apoptotic effect on sarcoma cell lines, sunitinib prompted PD-L1 upregulation on sarcoma cells. Interestingly, sunitinib-treated sarcoma cells drive DCs to full maturation and increase their capacity to induce sarcoma-reactive T cells to produce IFN-γ. Conversely, no effect on T cell proliferation and T cell subpopulation composition was observed. Moreover, both bone and synovial sarcoma cell lines induced Tregs through DCs but sunitinib treatment completely abrogated Treg induction. Finally, sarcoma cell lines induced PD-1 upregulation on both effector T cells and Tregs when loaded into DCs, providing a rationale for using PD-1 blockade. Indeed, PD-1 blockade by nivolumab synergized with sunitinib in inducing IFN-γ-producing effector T cells.

Conclusions

Taken together, our in vitro data indicate that the treatment of sarcoma cells with sunitinib can exert significant changes on immune cell subsets toward immune activation, leading to DC-based cross-priming of IFN-γ-producing effector T cells and reduced Treg induction. PD-1 blockade with nivolumab has a synergistic effect with sunitinib, supporting the use of TKI and anti-PD-1 approach in sarcomas, and perhaps in other cancers. DC-targeted drugs, including toll-like receptor 3 inhibitors and CD47 inhibitors, are under development and our preclinical model might help to better design their clinical application.
Autoimmune Addison's Disease as Part of the Autoimmune Polyglandular Syndrome Type 1: Historical Overview and Current Evidence
Authors: Perniola Roberto , Fierabracci Alessandra , Falorni Alberto
Published: February 2021

The autoimmune polyglandular syndrome type 1 (APS1) is caused by pathogenic variants of the autoimmune regulator (AIRE) gene, located in the chromosomal region 21q22.3. The related protein, AIRE, enhances thymic self-representation and immune self-tolerance by localization to chromatin and anchorage to multimolecular complexes involved in the initiation and post-initiation events of tissue-specific antigen-encoding gene transcription. Once synthesized, the self-antigens are presented to, and cause deletion of, the self-reactive thymocyte clones. The clinical diagnosis of APS1 is based on the classic triad idiopathic hypoparathyroidism (HPT)—chronic mucocutaneous candidiasis—autoimmune Addison's disease (AAD), though new criteria based on early non-endocrine manifestations have been proposed. HPT is in most cases the first endocrine component of the syndrome; however, APS1-associated AAD has received the most accurate biochemical, clinical, and immunological characterization. Here is a comprehensive review of the studies on APS1-associated AAD from initial case reports to the most recent scientific findings.
Combining IL-10 and Oncolytic Adenovirus Demonstrates Enhanced Antitumor Efficacy Through CD8+ T Cells
Authors: Chen Duo , Huang Luyu , Zhou Haiyu , Zhang Yuhui
Published: February 2021

Oncolytic viruses are of growing importance in cancer therapeutics since they combine direct oncolytic effect and the stimulation of antitumor immunity. Emerging evidences showed that the function of oncolytic viruses is dependent on immune response in tumor microenvironment, and the modulation of immunity could influence their efficacy. Here we combined the interleukin 10 (IL-10) and oncolytic adenovirus Ad-hTERT to treat lung cancer and explored the underlying mechanism under combination therapy. Lewis lung carcinoma (LLC) and B16F10 tumor-bearing immunocompetent C57BL/6 mice that received Ad-hTERT or IL-10 alone showed mild antitumor effect, while the combination therapy shrink tumor bulks and prolonged survival remarkably. In addition, IL-10 didn’t show direct influence on tumor cell viability or Ad-hTERT mediated tumor cell lysis in vitro. To further explore the influence of combination therapy mediated antitumor capacity, we eliminated CD8+ T, CD4+ T or natural killer (NK) cells in LLC and B16F10-bearing C57BL/6 mice, and found that CD8+ T cells were critical mediator in the combination therapy. The combination therapy induced intensive infiltration of CD8+ T cells in tumors, increased tumor-specific IFN-γ secretion by CD8+ T cells. The long-term tumor-specific immune memory induced by the combination therapy rejected rechallenge by respective tumor cell lines. This study demonstrated that the therapy combining IL-10 and Ad-hTERT augmented antitumor efficacy which was CD8+ T cells dependent. Our findings paved the way to combine cytokines and oncolytic viruses to enhance antitumor immunotherapy in treating cancer.
The Impact of Tregs on the Anticancer Immunity and the Efficacy of Immune Checkpoint Inhibitor Therapies
Authors: González-Navajas Jose M. , Fan Dengxia Denise , Yang Shuang , Yang Fengyuan Mandy , Lozano-Ruiz Beatriz , Shen Liya , Lee Jongdae
Published: February 2021

Although cancers arise from genetic mutations enabling cells to proliferate uncontrollably, they cannot thrive without failure of the anticancer immunity due in a large part to the tumor environment's influence on effector and regulatory T cells. The field of immune checkpoint inhibitor (ICI) therapy for cancer was born out of the fact that tumor environments paralyze the immune cells that are supposed to clear them by activating the immune checkpoint molecules such as PD-1. While various subsets of effector T cells work collaboratively to eliminate cancers, Tregs enriched in the tumor environment can suppress not only the native anticancer immunity but also diminish the efficacy of ICI therapies. Because of their essential role in suppressing autoimmunity, various attempts to specifically deplete tumor-associated Tregs are currently underway to boost the efficacy of ICI therapies without causing systemic autoimmune responses. A better understanding the roles of Tregs in the anti-cancer immunity and ICI therapies should provide more specific targets to deplete intratumoral Tregs. Here, we review the current understanding on how Tregs inhibit the anti-cancer immunity and ICI therapies as well as the advances in the targeted depletion of intratumoral Tregs.
Epigenetic Landscapes of Single-Cell Chromatin Accessibility and Transcriptomic Immune Profiles of T Cells in COVID-19 Patients
Authors: Li Shun , Wu Bin , Ling Yun , Guo Mingquan , Qin Boyin , Ren Xiaonan , Wang Chao , Yang Hua , Chen Lixiang , Liao Yixin , Liu Yang , Peng Xiuhua , Xu Chunhua , Wang Zhenyan , Shen Yinzhong , Chen Jun , Liu Li , Niu Bowen , Zhu Mengmin , Liu Lingling , Li Feng , Zhu Tongyu , Zhu Zhaoqin , Zhou Xiaohui , Lu Hongzhou
Published: February 2021

T cells play a critical role in coronavirus diseases. How they do so in COVID-19 may be revealed by analyzing the epigenetic chromatin accessibility of cis- and trans-regulatory elements and creating transcriptomic immune profiles. We performed single-cell assay for transposase-accessible chromatin (scATAC) and single-cell RNA (scRNA) sequencing (seq) on the peripheral blood mononuclear cells (PBMCs) of severely ill/critical patients (SCPs) infected with COVID-19, moderate patients (MPs), and healthy volunteer controls (HCs). About 76,570 and 107,862 single cells were used, respectively, for analyzing the characteristics of chromatin accessibility and transcriptomic immune profiles by the application of scATAC-seq (nine cases) and scRNA-seq (15 cases). The scATAC-seq detected 28,535 different peaks in the three groups; among these peaks, 41.6 and 10.7% were located in the promoter and enhancer regions, respectively. Compared to HCs, among the peak-located genes in the total T cells and its subsets, CD4+ T and CD8+ T cells, from SCPs and MPs were enriched with inflammatory pathways, such as mitogen-activated protein kinase (MAPK) signaling pathway and tumor necrosis factor (TNF) signaling pathway. The motifs of TBX21 were less accessible in the CD4+ T cells of SCPs compared with those in MPs. Furthermore, the scRNA-seq showed that the proportion of T cells, especially the CD4+ T cells, was decreased in SCPs and MPs compared with those in HCs. Transcriptomic results revealed that histone-related genes, and inflammatory genes, such as NFKBIA, S100A9, and PIK3R1, were highly expressed in the total T cells, CD4+ T and CD8+ T cells, both in the cases of SCPs and MPs. In the CD4+ T cells, decreased T helper-1 (Th1) cells were observed in SCPs and MPs. In the CD8+T cells, activation markers, such as CD69 and HLA class II genes (HLA-DRA, HLA-DRB1, and HLA-DRB5), were significantly upregulated in SCPs. An integrated analysis of the data from scATAC-seq and scRNA-seq showed some consistency between the approaches. Cumulatively, we have generated a landscape of chromatin epigenetic status and transcriptomic immune profiles of T cells in patients with COVID-19. This has provided a deeper dissection of the characteristics of the T cells involved at a higher resolution than from previously obtained data merely by the scRNA-seq analysis. Our data led us to suggest that the T-cell inflammatory states accompanied with defective functions in the CD4+ T cells of SCPs may be the key factors for determining the pathogenesis of and recovery from COVID-19.
Acute Rheumatic Fever: Where Do We Stand? An Epidemiological Study in Northern Italy
Authors: Marino Achille , Cimaz Rolando , Pelagatti Maria Antonietta , Tattesi Giulia , Biondi Andrea , Menni Laura , Sala Marco , Calzi Patrizia , Morandi Francesco , Cortinovis Francesca , Cogliardi Anna , Addis Claudia , Bellù Roberto , Andreotti Massimo , Varisco Tiziana
Published: February 2021

Acute rheumatic fever (ARF) is a non-septic complication of group A β-hemolytic streptococcal (GAS) throat infection. Since 1944, ARF diagnosis relies on the Jones criteria, which were periodically revised. The 2015 revision of Jones criteria underlines the importance of knowing the epidemiological status of its own region with updated data. This study aims to describe ARF features in a retrospective cohort retrieved over a 10-year timespan (2009–2018) and to report the annual incidence of ARF among children in the Province of Monza-Brianza, Lombardy, Italy during the same period. This is a multicentric cross-sectional/retrospective study; 70 patients (39 boys) were diagnosed with ARF. The median age at diagnosis was 8.5 years (range, 4–14.2 years). Overall, carditis represented the most reported major Jones criteria followed by arthritis and chorea (40, 27, and 20 cases, respectively). In order to calculate the annual incidence of ARF, only children resident in the Province of Monza-Brianza were included in this part of the analysis. Therefore, 47 patients aged between 5 and 14 years were identified. The median incidence during the study time was 5.7/100,000 (range, 2.8–8.3/100,000). In the Province of Monza-Brianza, we found an incidence rate of ARF among children aged 5–14 years constantly above the threshold of low-risk area as defined in the 2015 revision of Jones criteria. Therefore, the diagnosis of ARF should be based on the moderate–high-risk set of Jones criteria. However, given the burden of secondary prophylaxis, expert opinion is advisable when the diagnosis of ARF is uncertain.
Loss of Basal Forebrain Cholinergic Neurons Following Adolescent Binge Ethanol Exposure: Recovery With the Cholinesterase Inhibitor Galantamine
Authors: Crews Fulton T. , Fisher Rachael , Deason Chloe , Vetreno Ryan P.
Published: February 2021

Binge drinking and alcohol abuse are common during adolescence and cause both cognitive deficits and lasting cholinergic pathology in the adult basal forebrain. Acetylcholine is anti-inflammatory and studies using the preclinical adolescent intermittent ethanol (AIE; 5.0 g/kg, i.g., 2 day on/2 day off from postnatal day [P]25 to P54) model of human adolescent binge drinking report decreased basal forebrain cholinergic neurons (BFCNs) and induction of proinflammatory genes that persist long into adulthood. Recent studies link AIE-induced neuroimmune activation to cholinergic pathology, but the underlying mechanisms contributing to the persistent loss of BFCNs are unknown. We report that treatment with the cholinesterase inhibitor galantamine (4.0 mg/kg, i.p.) administered during AIE (i.e., P25–P54) or following the conclusion of AIE (i.e., P57–P72) recovered the persistent loss of cholinergic neuron phenotype markers (i.e., ChAT, TrkA, and p75NTR) and somal shrinkage of residual ChAT + neurons known to persist in AIE-exposed adults. Galantamine treatment also recovered the AIE-increased expression of the proinflammatory receptors TLR4 and RAGE, the endogenous TLR4/RAGE agonist HMGB1, and the transcription activation marker pNF-κB p65. Interestingly, we find BFCNs express TLR4 and RAGE, and that AIE treatment increased pNF-κB p65 expression in adult ChAT + IR neurons, consistent with intracellular HMGB1-TLR4/RAGE signaling within BFCNs. AIE increased epigenetic transcription silencing markers (i.e., H3K9me2 and H3K9me3) in the adult basal forebrain and H3K9me2 occupancy at cholinergic phenotype gene promoters (i.e., ChAT and TrkA). The finding of no AIE-induced changes in total basal forebrain NeuN + neurons with galantamine reversal of AIE-induced ChAT + neuron loss, TLR4/RAGE-pNF-κB p65 signals, and epigenetic transcription silencing markers suggests that AIE does not cause cell death, but rather the loss of the cholinergic phenotype. Together, these data suggest that AIE induces HMGB1-TLR4/RAGE-pNF-κB p65 signals, causing the loss of cholinergic phenotype (i.e., ChAT, TrkA, and p75NTR) through epigenetic histone transcription silencing that result in the loss of the BFCN phenotype that can be prevented and restored by galantamine.
Quantitative Fluorescent in situ Hybridization Reveals Differential Transcription Profile Sharpening of Endocytic Proteins in Cochlear Hair Cells Upon Maturation
Authors: Huang Guobin , Eckrich Stephanie
Published: February 2021

The organ of Corti (OC) comprises two types of sensory cells: outer hair cells (OHCs) and inner hair cells (IHCs). While both are mechanotransducers, OHCs serve as cochlear amplifiers, whereas IHCs transform sound into transmitter release. Reliable sound encoding is ensured by indefatigable exocytosis of synaptic vesicles associated with efficient replenishment of the vesicle pool. Vesicle reformation requires retrieval of vesicle membrane from the hair cell’s membrane via endocytosis. So far, the protein machinery for endocytosis in pre-mature and terminally differentiated hair cells has only partially been deciphered. Here, we studied three endocytic proteins, dynamin-1, dynamin-3, and endophilin-A1, by assessing their transcription profiles in pre-mature and mature mouse OCs. State-of-the-art RNAscope® fluorescent in situ hybridization (FISH) of whole-mount OCs was used for quantification of target mRNAs on single-cell level. We found that pre-mature IHCs contained more mRNA transcripts of dnm1 (encoding dynamin-1) and sh3gl2 (endophilin-A1), but less of dnm3 (dynamin-3) than OHCs. These differential transcription profiles between OHCs and IHCs were sharpened upon maturation. It is noteworthy that low but heterogeneous signal numbers were found between individual negative controls, which highlights the importance of corresponding analyses in RNAscope® assays. Complementary immunolabeling revealed strong expression of dynamin-1 in the soma of mature IHCs, which was much weaker in pre-mature IHCs. By contrast, dynamin-3 was predominantly found in the soma and at the border of the cuticular plates of pre-mature and mature OHCs. In summary, using quantitative RNAscope® FISH and immunohistochemistry on whole-mount tissue of both pre-mature and mature OCs, we disclosed the cellular upregulation of endocytic proteins at the level of transcription/translation during terminal differentiation of the OC. Dynamin-1 and endophilin-A1 likely contribute to the strengthening of the endocytic machinery in IHCs after the onset of hearing, whereas expression of dynamin-3 at the cuticular plate of pre-mature and mature OHCs suggests its possible involvement in activity-independent apical endocytosis.
A Retrospective Longitudinal Study in a Cohort of Children With Dyskinetic Cerebral Palsy Treated With Tetrabenazine
Authors: Scalise Roberta , Sgandurra Giuseppina , Menici Valentina , Capodagli Nicola , Di Pietro Roberta , Romeo Domenico M. , Sini Francesca , Pagliano Emanuela , Foscan Maria , Cioni Giovanni , Battini Roberta
Published: February 2021

Tetrabenazine has been studied with a variety of hyperkinetic movement disorders, but there is limited and empirical literature on the potential efficacy of tetrabenazine in children with dyskinetic cerebral palsy (DCP). The purpose of this study was to evaluate the efficacy of tetrabenazine in a sample of children with DCP using the Movement Disorders—Childhood Rating Scale 4–18 Revised (MD-CRS 4–18 R). The study is a multicenter retrospective longitudinal study in which the participants were selected from the databases of each Center involved, according to detailed inclusion criteria. The study was performed on 23 children and adolescents (19 male and 4 females; mean age 8.28 years, SD 3.59) with DCP having been evaluated before starting the treatment (baseline), after 6 and 12 months of treatment and in a sub-cohort after >2 years follow-up. A linear mixed model was used to evaluate the effects of the different timings on each MD-CRS 4–18 R Index (Index I, Index II, and Global Index) adding age and type of movement disorder as random effect. A significant clinical improvement related to a reduction of MD-CRS 4-18 R Indexes was detected between the baseline and after 6 and 12 months of treatment. Findings support the efficacy of tetrabenazine in children with DCP through a standardized outcome measure (MD-CRS 4–18 R) and confirm the use of this scale as a suitable tool to detect changes in further randomized clinical trials.
Case Report: Severe Osteoporosis and Preventive Therapy in RNA Polymerase III-Related Leukodystrophy
Authors: Furukawa Soma , Kunii Misako , Doi Hiroshi , Kondo Naohide , Ogura Aya , Hirabuki Koichi , Itoh Takayuki , Matsumoto Naomichi , Tanaka Fumiaki , Katsuno Masahisa , Ito Yasuhiro
Published: February 2021

RNA polymerase III (POLR3)-related leukodystrophy is an autosomal recessive form of leukodystrophy caused by homozygous or compound heterozygous mutations of the RNA polymerase III subunit genes, including subunit A (POLR3A). With respect to the manifestation triad, hypomyelination, hypodontia, and hypogonadotropic hypogonadism, it is also known as 4H leukodystrophy. Here, we report a 41-year-old woman of POLR3-related leukodystrophy by carrying compound heterozygous pathogenic variants of c.2554A>G (p.M852V) and c.2668G>T (p.V890F) in the POLR3A gene. She was amenorrheic and became a wheelchair user from the age of 15 years and suffered from multiple episodes of pathologic fractures, starting with a subtrochanteric fracture of the right femur after a tonic seizure at age 30 years. Head magnetic resonance imaging demonstrated hypomyelination and atrophies of the cerebellum, brainstem, and corpus callosum. Laboratory examination revealed a marked decrease of gonadotropins and estrogen, low bone density, and high bone resorption markers. Administration of anti-receptor activator of nuclear factor kappa-B ligand monoclonal antibody restored bone resorption markers to a normal level and prevented further pathological bone fractures. Our case emphasizes that osteoporosis should be recognized as a potential but serious complication in POLR3-related leukodystrophy. It may be feasible to prevent pathologic fractures by intensive osteoporosis therapy after endocrinological examinations and evaluation of bone metabolism.
EEG Power Spectral Analysis of Abnormal Cortical Activations During REM/NREM Sleep in Obstructive Sleep Apnea
Authors: Liu Shuling , Shen Jiucheng , Li Yezhou , Wang Jing , Wang Jianhua , Xu Juan , Wang Qiaojun , Chen Rui
Published: February 2021

Objective: To characterize electroencephalogram (EEG) power in different frequency bands during rapid eye movement (REM) sleep and non-rapid eye movement (NREM) sleep in patients with obstructive sleep apnea (OSA).

Methods: Retrospective data on 151 patients were collected and divided into three groups: primary snoring group (AHI < 5/h), mild-moderate OSA group (6 ≤ AHI < 30/h), and severe OSA group (AHI ≥ 30/h). EEG recordings in the frontal, central, and occipital regions were extracted from both REM and NREM sleep, to compute the normalized spectral power densities in the delta, theta, alpha, sigma, beta, and gamma frequency bands, using Fast Fourier Transform. Correlations between the computed EEG power and PSG parameters were analyzed.

Results: In NREM sleep, elevated normalized power spectral density (PSD) in the delta band was observed in the severe OSA group compared to the other two groups. In contrast, the PSD of the other frequency bands showed a corresponding decrease in the severe OSA group. In REM sleep, similar changes were observed in the frontal region. Delta band PSD was positively correlated with Apnea Hypopnea Index (AHI) (r = 0.33), longest time of apnea, oxygen desaturation index (ODI) (r = 0.34), percent sleep time below 90% SaO2 (T90%) (r = 0.30), Arousal Index (ArI) (r = 0.29), and negatively correlated with N3%, minimum oxygen saturation (minSaO2).

Conclusion: Our findings provide neurophysiological evidence for pathological cortical activation during REM/NREM sleep, which may be associated with the arousals and cognitive impairments in OSA. The technique of power spectral analysis could prove a potentially useful tool in complementing traditional PSG parameters in assessing disease burden to guide therapeutic decisions.
The Body Across Adulthood: On the Relation Between Interoception and Body Representations
Authors: Raimo Simona , Boccia Maddalena , Di Vita Antonella , Cropano Maria , Guariglia Cecilia , Grossi Dario , Palermo Liana
Published: February 2021

Interoceptive information plays a pivotal role in building body representations (BR), but the association between interoception and the different types of BR in healthy individuals has never been systematically investigated. Thus, this study aimed to explore the association between BR and interoceptive sensibility (IS) throughout adulthood. One hundred thirty-seven healthy participants (50 aged from 18 to 40 years old; 50 aged from 41 to 60 years old; and 37 over 60 years old) were given a self-report tool for assessing IS (the Self-Awareness Questionnaire; SAQ), and a specific battery including tasks evaluating three different BR (i.e., the body schema, using the Hand Laterality Task; the body structural representation, using the Frontal Body Evocation task, FBE; and body semantics, using the Object-Body Part Association Task) as well as control tasks (i.e., tasks with non-body stimuli). The older age group (aged over 60 years old) showed lower performances on the tasks probing the body schema and body structural representation than younger groups (aged 18 to 40 and 41 to 60 years old). More interestingly, worse performances on a task assessing the body schema were significantly associated with higher IS with older age, suggesting that higher awareness of one’s inner body sensations would decrease the plasticity of this BR. These findings are interpreted according to the neuropsychological model of BR development and the effects of aging on the brain.
Voltage-Gated Ca2+ Channels in Dopaminergic Substantia Nigra Neurons: Therapeutic Targets for Neuroprotection in Parkinson's Disease?
Authors: Ortner Nadine J.
Published: February 2021

The loss of dopamine (DA)-producing neurons in the substantia nigra pars compacta (SN) underlies the core motor symptoms of the progressive movement disorder Parkinson's disease (PD). To date, no treatment to prevent or slow SN DA neurodegeneration exists; thus, the identification of the underlying factors contributing to the high vulnerability of these neurons represents the basis for the development of novel therapies. Disrupted Ca2+ homeostasis and mitochondrial dysfunction seem to be key players in the pathophysiology of PD. The autonomous pacemaker activity of SN DA neurons, in combination with low cytosolic Ca2+ buffering, leads to large somatodendritic fluctuations of intracellular Ca2+ levels that are linked to elevated mitochondrial oxidant stress. L-type voltage-gated Ca2+ channels (LTCCs) contribute to these Ca2+ oscillations in dendrites, and LTCC inhibition was beneficial in cellular and in vivo animal models of PD. However, in a recently completed phase 3 clinical trial, the dihydropyridine (DHP) LTCC inhibitor isradipine failed to slow disease progression in early PD patients, questioning the feasibility of DHPs for PD therapy. Novel evidence also suggests that R- and T-type Ca2+ channels (RTCCs and TTCCs, respectively) represent potential PD drug targets. This short review aims to (re)evaluate the therapeutic potential of LTCC, RTCC, and TTCC inhibition in light of novel preclinical and clinical data and the feasibility of available Ca2+ channel blockers to modify PD disease progression. I also summarize their cell-specific roles for SN DA neuron function and describe how their gating properties allow activity (and thus their contribution to stressful Ca2+ oscillations) during pacemaking.