Articles de revues sur le sujet « Gunn mouse »

Heme oxygenase-1 (HO-1) is induced in the vasculature in the DOCA-salt model of hypertension in rats. Whereas the HO system and its products may exert vasodilator effects, recent studies have suggested that the HO system may predispose to hypertension. The present study examined the effects of selected components of the HO system, specifically, the HO-1 isozyme and the product bilirubin in the DOCA-salt model of systemic hypertension; the experimental approach employed mutant rodent models, namely, the HO-1−/− mouse and the hyperbilirubinemic Gunn rat. DOCA-salt induced HO-1 protein in the aorta in HO-1+/+ mice and provoked a significant rise in systolic arterial pressure in HO-1−/− mice but not in HO-1+/+ mice; this effect could not be ascribed to impaired urinary sodium excretion or impaired glomerular filtration rate in the DOCA-salt-treated HO-1−/− mice. The administration of DOCA salt to uninephrectomized rats significantly increased systolic arterial pressure in wild-type rats, an effect that was attenuated in the mutant Gunn rat; this reduction in systemic hypertension in the DOCA-salt-treated Gunn rat was not due to a greater induction of HO-1 in the vasculature or to a more avid urinary sodium excretion. DOCA-salt impaired endothelium-dependent and endothelium-independent vasorelaxation in wild-type rats but not in Gunn rats; prior exposure to bilirubin repaired the defect in endothelium-dependent vasorelaxation in aortic rings in DOCA-salt-treated rats. DOCA salt stimulated vascular production of superoxide anion in wild-type but not in Gunn rats. We suggest that HO-1 and the product bilirubin may exert a countervailing effect in the DOCA-salt model of systemic hypertension.

ABSTRACT Sordarin derivatives constitute a new group of synthetic antifungal agents that selectively inhibit fungal protein synthesis. They have demonstrated in vitro activity against the most important fungal pathogens, both yeast and filamentous. This new family of compounds has also shown in vivo activity against murineCandida albicans, Histoplasma capsulatum, andCoccidioides immitis experimental infections, as well as against Pneumocystis carinii pneumonia in rats. After intravenous dosing in animals, both the area under the concentration-time curve and the elimination half-life were highest in Cynomolgus monkeys, followed by those in rats, mice, and rabbits. The volume of distribution at steady state for sordarin derivatives was similar in all species tested. The clearance in rats and mice was higher than for other species. GM 237354, a sordarin derivative, was characterized by high serum protein binding in mouse, rat, and monkey serum (unbound fraction, ≤5%). An indirect evaluation of the effect of liver function upon the metabolism of this class of compounds has been made in animals with impaired liver function such as Gunn rats, as well as in allometric studies that showed better correlations of half-life to liver blood flow than to animal body weight. Linearity of the main pharmacokinetic parameters was demonstrated after intravenous dosing of the representative compound GM 193663 at 10 and 20 mg/kg of body weight in rats. Allometry was used to determine whether human pharmacokinetic parameters can be predicted from animal data by regression analysis against body weight and liver blood flow. All these results have demonstrated that the human pharmacokinetics of sordarin derivatives can be forecast from animal data.

Abstract Major Histocompatibility Complex (MHC) Class I downregulation is a well described mechanism of tumor immune escape, posing a challenge for T cell based immunotherapies including immune checkpoint blockade (ICB). Recent studies, however, have demonstrated mixed roles of MHC Class 1 and the critical component beta-2-microglobulin (β2m) expression in cancer progression and ICB response, with some studies showing inactivation of antigen presentation to be associated with resistance to ICB and others showing low β2m expression to be associated with favorable prognosis. Glioblastoma (GBM) in particular expresses little or no MHC Class 1 and patients remain unresponsive to ICB. We thus sought to evaluate the role of MHC Class 1 in ICB, given that we have previously demonstrated that combination ICB with anti-PD-1 and co-stimulation with 4-1BB agonism has marked efficacy against intracranial murine glioma tumors in a CD8 T cell dependent manner. Surprisingly, in a CT2A murine glioma tumor line expressing the antigen TRP2 and lacking cell surface MHC I (CT2A-TRP2-β2mKO), the efficacy of combination 4-1BB and PD-1 therapy (ICB) was re-demonstrated in a CD8 dependent manner, independent of NK cells, CD4 T cells, and B cells. Furthermore, the efficacy of immunotherapy against intracranial CT2A-TRP2-β2mKO was demonstrated to be antigen dependent, with an adoptive lymphocyte transfer (ALT) of TRP2 TCR transduced T cells (TRP2 T cells) into a CD8KO mouse sufficient to eliminate CT2A-TRP2-β2mKO in the setting of ICB. Additionally, an ALT of TRP2 T cells did not kill CT2A-β2mKO tumors in the setting of ICB, while OT-1 mice whose CD8+ T cells primarily recognize OVA peptide with CT2A-TRP2-β2mKO tumors did not respond to ICB. In vitro studies revealed that TRP2 T cells demonstrated anti-tumor cytotoxicity against MHC Class I negative CT2A-TRP2-β2mKO tumor cells in the presence of TRP2 loaded bone marrow derived macrophages (TRP2 Mφ), but neither cell type was individually sufficient to induce tumor cell death, while the combination of TRP2 T cells and TRP2 Mφ demonstrated no cytotoxicity against CT2A-β2mKO tumors. Transwell experiments in which TRP2 Mφ and CT2A-TRP2-β2mKO tumor cells were separated by a 0.5µm membrane permeable to T cells but not Mφ or tumor cells revealed that contact between TRP2 Mφ and tumor cells was not necessary to induce T cell dependent killing. Indeed, tumor-bearing β2mKO bone marrow chimeras lacking MHC class 1 on hematopoeitically derived cells did not respond to ICB, highlighting the importance of antigen presentation from myeloid cells. The mechanism of killing was found to be dependent on interferon gamma (IFNγ), as IFNγKO mice did not respond to ICB. These findings demonstrate that tumors with low MHC Class 1 expression may still be targeted by T cell dependent immunotherapies such as ICB when antigen presentation can occur from myeloid cells. Citation Format: Emily Lerner, Vincent D'Anniballe, William Tomaszewski, Jonathan Perera, Xiuyu Cui, Jessica Waibl-Polania, Daniel Wilkinson, Michael D. Gunn, Peter E. Fecci, Karolina Woroniecka. CD8 T cell mediated killing of MHC class 1 negative tumors requires antigen presenting myeloid cells and interferon gamma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1378.

Sandi morse adalah suatu bentuk kode atau isyarat untuk berkomunikasi. Kode-kode ini biasanya dipakai di organisasi kepramukaan sebagai sarana pendidikan untuk mencari metode komunikasi yang lebih efektif digunakan di saat-saat tertentu. Pembelajaran sandi morse dapat menjadi sarana untuk melatih para anggota Pramuka agar menjadi terampil, cerdas, dan memiliki indera pendengaran atau penglihatan yang tajam. Dalam dunia Pramuka, sandi morse juga kerap dipelajari sebagai salah satu bentuk keterampilan. Kemampuan seorang anggota pramuka untuk mengirimkan dan menerima kode morse adalah salah satu dari kecakapan yang dapat menerima tanda kecakapan khusus. Dibalik sekian banyak manfaat yang dihasilkan dari sandi morse, banyak diantara kita punya kesulitan untuk menerjemahkan kode sandi morse karena sandi ini hanya terdiri dari simbol titik dan garis yang rumit, jika dilihat sekilas terasa mirip. Berdasarkan uraian di atas, tujuan pembuatan tugas akhir ini adalah untuk pengerjaan aplikasi berbasis kata sandi morse berdasarkan Android guna memudahkan pengguna menejermahkan kalimat ke bentuk sandi morse secara cepat dan akurat. Aplikasi ini tidak hanya bisa mengubah kode menjadi kode morse pelihara juga mampu mengubah kode morse menjadi kalimat. Diharapkan dengan aplikasi ini dapat membantu pengguna dalam menerjemahkan kalimat ke sandi morse or sebaliknya sehingga pengguna tidak akan mengalami kesulitan lagi ketika penerjemahan kode morse.

ABSTRACT p27Kip1 is a member of the Cip-Kip family of cyclin-dependent kinase (Cdk) inhibitors that binds to cyclin-Cdk complexes and inhibits their catalytic activity in response to antiproliferative stimuli. p27Kip1 is regulated by several posttranscriptional mechanisms, including subcellular localization. We have identified a component of the nuclear pore complex (NPC), termed Nup50, through its two-hybrid interactions with p27Kip1. Nup50 is a nucleoplasmically oriented component of the nuclear pore complex with a role in protein export (T. Guan, R. H. Kehlenbach, E. C. Schirmer, A. Kehlenbach, F. Fan, B. E. Clurman, N. Arnheim, and L. Gerace, Mol. Cell. Biol. 20:5619–5630, 2000). We found that murine Nup50 is a widely expressed nucleoporin and that Nup50 expression is highest in the developing neural tube and adult testes. We have also examined interactions between Nup50 and the NPC and found specific two-hybrid interactions between Nup50 and several well-defined components of the NPC, as well as coimmunoprecipitation of Nup50 with the nucleoporin Nup153 from transfected mammalian cells. In order to study Nup50 function in vivo, we cloned the mouse Nup50 genomic locus and created a targeted Nup50 deletion in the mouse germ line. Nup50 disruption resulted in a complex phenotype characterized by late embryonic lethality, neural tube defects, and intrauterine growth retardation. Although Nup50-null mouse embryo fibroblasts exhibited no defects in either cell cycle control or p27Kip1 regulation, Nup50 deletion was associated with abnormalities in p27Kip1expression and cell proliferation in the developing neuroepithelium. We conclude that Nup50 is a nucleoporin with essential functions during mouse development.

Phenotypic changes in C3H/10T mouse embryo fibroblast cells, induced by retinyl acetate (RAC) or 12-0-tetradecanoylphorbol-13-acetate (TPA) treatments, were studied by freeze fracture electron microscopy. TPA accelerates transformation while RAC suppresses transformation in these cells.Unfixed cells on glass substrates were rapidly frozen in liquid propane by the sandwich method. Freeze fracture was carried out at -120°C and vacuum better than 5x10-7torr in a Polaron unit equipped with a Perkin-Elmer Ultek ion pump and Cressi ngton electron guns. Intramembraneous particle (IMP) detection and quantitation was done using a Cambridge Instrument Quantimet 920 image analyzer with a macroviewer attachment.

The goal of understanding fall risk in the elderly, prevention and protection is to improve clinical and care satisfaction. Another anticipatory method that can be used to predict falling conditions is the assessment of the risk of falling in the elderly. The Hendrich Fall Scale (HFS) and Morse Fall Scale (MFS) are a form of assessment to anticipate the risk of falling in the elderly in nursing homes for patients. The aim is to determine the effectiveness of the Hendrich Fall Scale and the Morse Fall Scale with an assessment of the Risk of Fall in the Elderly. The research design used in this study is a longitudinal comparative design. The total sample in this study was 40 elderly. This research was conducted at the Nursing Home Foundation Guna Budi Bakti Medan Labuhan. Data collection using the Hendrich Fall Scale and Morse Fall Scale. Data analysis using Chi Square. Fall risk assessment using the Hendrich Fall Scale (HFS), elderly people with a high risk of falling (25.0%), moderate risk of falling (65.0%). Fall risk assessment used the Morse Fall Scale (MFS), the elderly who had a high risk of falling (39.1%), moderate risk of falling (47.8%). It is recommended that seniors at risk of falling should be assessed using the MFS instrument.

Guillot, Pascale V., Lixin Liu, Jan Albert Kuivenhoven, Jason Guan, Robert D. Rosenberg, and William C. Aird. Targeting of human eNOS promoter to the Hprt locus of mice leads to tissue-restricted transgene expression. Physiol Genomics 2: 77–83, 2000.—Phenotypic heterogeneity of the endothelium arises from cell type-specific differences in gene expression. An understanding of the mechanisms that underlie differential gene expression would provide important insight into the molecular basis of vascular diversity. In standard transgenic assays, multiple copies of heterologous DNA cassettes are randomly integrated into the mouse genome, resulting in significant line-to-line variation in expression. To overcome these limitations, we have targeted a single copy of a transgene that contains 1,600 bp of the human endothelial nitric oxide synthase (eNOS) promoter coupled to the LacZ reporter gene to the X-linked hypoxanthine phosphoribosyltransferase ( Hprt) locus of mice by homologous recombination. The transgene was inserted in either of the orientations relative to that of the Hprt gene. In mice derived from multiple embryonic stem (ES) cell clones, the expression pattern was limited to a subset of endothelial cells, cardiomyocytes, and vascular smooth muscle cells. These findings suggest that Hprt locus targeting is a feasible tool for studying endothelial cell-restricted gene regulation.

Abstract Cytokines are potent stimulators of the immune system and have long been investigated and used as therapeutics to treat cancer. However, free cytokines have short half-life and the lack of tumor targeting often results in systemic toxicity. IL-15 is a stimulatory cytokine that shares the common beta and gamma receptors with IL-2. In contrast to IL-2, IL-15 is more selective for NK and effector memory T cells, with less proliferative effect on Tregs, making it a more preferred therapeutic candidate for cancer. QL415 is an anti-PD-L1 x IL-15 fusion protein designed to enhance tumor accumulation and prolong blood circulation, thereby widening the therapeutic window. IL-15 and IL-15 receptor alpha sushi domain are covalently linked to the C-terminus of our proprietary PD-L1 antibody. Fc-mediated effector functions were ablated using mutations in the CH2 domain, to prevent depletion of effector immune cells. In vitro studies showed QL415 to be highly potent in promoting proliferation and pSTAT5 signaling of IL-15 -responding NK92 and M07e cells. QL415 selectively induced proliferation of NK and CD8+ T cells from human PBMC in vitro and mouse CD8+ and NK1.1 cells in vivo. In a MC38 mouse colon cancer model expressing human PD-L1, QL401 was highly effective in suppressing tumor growth, in contrast to PD-L1 monoclonal antibody or a non-targeted IL-15 control. Follow up tumor rechallenge study showed protective memory against cognate MC38 but not B16F10. In a separate MC38 tumor model using a mouse surrogate molecule of QL415, immunophenotyping of antigen presenting cells offered preliminary evidence of enrichment of conventional dendritic cells 1 in the tumor draining lymph nodes. Therefore, QL415 not only directly stimulates effector immune cells, but also indirectly through antigen presenting cells, promotes robust anti-tumor response. In non-human primates, Q415 was tolerated up to 1.5 mg/kg, well above the effective therapeutic dose. In light of these favorable preclinical efficacy and safety results, a phase 1 dose escalation and expansion study has been initiated to evaluate the safety, tolerability, and early efficacy of QL415. Citation Format: Irene Tang, Lauren Schwimmer, Monica Jin, Shenda Gu, Wei Wei Prior, Arianne Capacio, Hieu V. Tran, Allan Chan, Anna McClain, Shihao Chen, Chuanzeng Cao, Xiaoran Wu, Yanling Xu, Dongmei Guan, Xi Chen, Xianli Su. QL415, a tumor targeted IL-15 fusion protein stimulating both lymphoid and myeloid immune cells for optimal anti-tumor immune response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3504.

The action potential (AP) is a fundamental feature of excitable cells that serves as the basis for long-distance signaling in the nervous system. There is considerable diversity in the appearance of APs and the underlying repolarization mechanisms in different neuronal types (reviewed in Bean BP. Nat Rev Neurosci 8: 451–465, 2007), including among pyramidal cell subtypes. In the present work, we used specific pharmacological blockers to test for contributions of Kv1, Kv2, or Kv4 channels to repolarization of single APs in two genetically defined subpopulations of pyramidal cells in layer 5 of mouse somatosensory cortex ( etv1 and glt) as well as pyramidal cells from layer 2/3. These three subtypes differ in AP properties (Groh A, Meyer HS, Schmidt EF, Heintz N, Sakmann B, Krieger P. Cereb Cortex 20: 826–836, 2010; Guan D, Armstrong WE, Foehring RC. J Neurophysiol 113: 2014–2032, 2015) as well as laminar position, morphology, and projection targets. We asked what the roles of Kv1, Kv2, and Kv4 channels are in AP repolarization and whether the underlying mechanisms are pyramidal cell subtype dependent. We found that Kv4 channels are critically involved in repolarizing neocortical pyramidal cells. There are also pyramidal cell subtype-specific differences in the role for Kv1 channels. Only Kv4 channels were involved in repolarizing the narrow APs of glt cells. In contrast, in etv1 cells and layer 2/3 cells, the broader APs are partially repolarized by Kv1 channels in addition to Kv4 channels. Consistent with their activation in the subthreshold range, Kv1 channels also regulate AP voltage threshold in all pyramidal cell subtypes.

Abstract Background: The microenvironment of nasopharyngeal carcinoma (NPC) has an intense and heterogenous immune infiltration, due to chronic EBV infection and locoregional lymphoid structures. Phase II clinical trials have shown that the response rate of NPC patients to PD-1 inhibitors is only 20%, indicating the current immunotherapies are not optimal due to insufficient understanding of the tumor-immune interaction. In this study, we have applied an integrated transcriptome sequencing analysis to decipher immune dynamics, and have characterized a new target that can inhibit Treg-mediated suppression in NPC, and provided critical translational evidence to the precision immunotherapy. Methods: We established an integrated NPC single-cell sequencing cohort containing 189,750 T cells from 50 patient samples. Spatial transcriptome sequencing was performed on frozen samples from eight NPC patients. The whole blood was collected from healthy donors and PBMCs were isolated using density gradient centrifugation. T cells were magnetically isolated from PBMCs. The humanized mouse model was established by tail-vein injection of CD3/CD28 activated PBMCs. Results: Single-cell sequencing revealed the enrichment of suppressive Tregs in the NPC microenvironment. We found that the C666-1 cell line facilitated Treg differentiation and activation. Spatial sequencing identified CD70-CD27 interaction was upregulated between NPC cells and suppressive Tregs, indicating its role in regulating Treg function. CD70 was predominantly expressed on NPC cells and correlated to worse prognosis. CD70-knockout (KO) in C666-1 inhibited Treg differentiation. Decreased Treg activities further enhanced the cytotoxicity in CD8+ effector T cells, leading to higher tumor apoptosis. Cusatuzumab also generated a comparable inhibitory effect in Treg-mediated suppression. In the humanized mouse model, CD70-KO and inhibition shrank the size of subcutaneous tumors by enhancing T cell immunity. Conclusion: The enrichment and activation Tregs makes the NPC microenvironment highly immunosuppressive, which might jeopardize the clinical efficacy of anti-PD1 therapies in NPC patients. In this study, we revealed that CD70+ NPC cells were highly resilient to immune surveillance since they were capable of modulating Treg differentiation and activation via the CD70-CD27 interaction to counteract CD8+ T cells. Blocking CD70 has been shown to effectively induce tumor apoptosis by alleviating Treg-mediated suppression. The therapeutic efficacy and safety of the anti-CD70 therapy were tested in the humanized mouse model. These findings suggested that the anti-CD70 therapy exhibited a promising efficacy to kill NPC cells by deteriorating the immunosuppressive microenvironment, and it might synergistically enhance the efficacy of anti-PD1 therapies. Citation Format: Lanqi Gong, Jie Luo, Yuma Yang, Yu Zhang, Xin-Yuan Guan. Single-cell spatial transcriptome sequencing characterizes CD70+ immune-resilient nasopharyngeal carcinoma cells as a vital mediator of Treg-induced suppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6127.

Abstract Prostate cancer is the second leading cause of cancer associated mortality in men, and for reasons that remain unclear, immunotherapy treatments have largely failed in patients with advanced prostate cancer. Standard of care therapy for these patients relies on androgen deprivation therapies (ADT) to block testosterone production combined with small molecules that inhibit androgen synthesis and/or the function of the androgen receptor (AR). Importantly, androgens are reported to be immunosuppressive; therefore, we postulated that androgen axis blockade might promote response to immunotherapy. In the recent years, the co-inhibitory receptor TIGIT has emerged as a promising immunotherapy target, and we observed that TIGIT is highly expressed on tumor infiltrating T cells and NK cells in metastatic castration resistant prostate cancer (CRPC) patients treated with the AR antagonist enzalutamide. Likewise, in a mouse model of CRPC, we show that tumor infiltrating T cells express TIGIT and that treatment with enzalutamide increases its expression. In this model, neither treatment with anti-TIGIT alone nor with chemical castration and enzalutamide affects tumor growth. Remarkably, the combination of anti-TIGIT with chemical castration and enzalutamide leads to delayed tumor growth and long-term cures in about 60% of the animals. Taken together, our data suggest that androgen receptor blockade sensitizes immune cells in the tumor to respond to anti-TIGIT immunotherapy. Citation Format: Fanny Polesso, Xiangnan Guan, Zheng Xia, Amy E. Moran. Androgen deprivation therapy allows for effective anti-TIGIT immunotherapy in murine model of castration resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 661.

Recent developments in Scanning Electron Microscopy have been technology driven. Major improvements have been made in information exchange, image processing technology, mechanical precision devices such as specimen stages, detector design, field emision guns, etc. In this process of automation and refining , the SEM became a rather complicated instrument to handle up to its full capabilities. When designing a user interface for such an instrument, considerable care must be taken that the instrument can be operated in an easy, straightforward way without losing functionality due to oversimplified automation.In the Philips XL SEM series, the user interface provides functional control over the microscope, where a function is defined by what the user wants to achieve, rather than by the individual hardware parts that have to be addressed in order to achieve the result. Such a function-based user interface can be realised by emploing a three-layered machine structure.The first layer consists of a mix of hardware parts such as an electron gun, lenses, a scan generator, an image processor, etc. In a second layer automation and compensation routines are performed by dedicated computers linked within a distributed intelligence network. Thus any of the primary parameters can be changed and the effect can be observed immediately as the related parameters are automatically readjusted. The third, outer layer is the actual user interface, implemented as customer-defined application programs running in an MS-DOS (MICROSOFT™) Windows environment on a personal computer. Simple mouse control provides fast access to a variety of application programs such as microscope setup, imaging, specimen survey, EDX analysis, linewidth measurement, image analysis and image database management. The window applications are overlaid on the microscope image, since the image is the essence of microscopy.

Literary images and functions of death in sword and sorcery and mythopoeic fantasyThis article presents and compares methods of description of death in two primary variants of fantasy literature: sword and sorcery and mythopoeic fantasy. The focus is on works of the precursors of fantasy literature — Robert E. Howard Conan the Barbarian series and John R.R. Tolkien The Lord of the Rings, Silmarillion, and texts of authors who creatively developed two primary types of fantasy literature — Fritz Leiber Fafhrd and the Gray Mouser cycle and Ursula K. Le Guin Earthsea cycle.The analysis is divided into two parts. The first one describes methods of presentation of death and their functions in sword and sorcery literature. In this variant of fantasy many literary images of death can be found, which focus particularly on its biological aspects. The next part shows analogical elements in mythopoeic fantasy, where the descriptions of death are inspired by the medieval chansons de geste.The article shows important differences between methods of presenting of death in sword and sorcery and mythopoeic fantasy and between functions of death in this two primary types of fantasy literature. In sword and sorcery the descriptions of death have great importance in the adventure plot structure, because they are connected with activities and adventures of the main character. In mythopoeic fantasy the kind of a character’s death often shows moral condition of this character. Moreover, death in mythopoeic fantasy is important for the balance and stability of created world.

Abstract There are few effective therapeutic options for metastatic lung adenocarcinomas (LUADs) that lack actionable mutations in receptor tyrosine kinases, and there is an urgent need to develop a better understanding of the biological basis of LUAD growth and metastasis. Our group has developed mouse models of LUAD metastasis and found that LUAD metastasis is epithelial-to-mesenchymal transition (EMT)-dependent. EMT causes cancer cells to switch their axis of polarity from apical-basal to front-rear, which orients organelles and actinbased cytoskeletal structures in ways that facilitate purposeful cancer cell motility and metastasis. The EMT-activating transcription factor ZEB1 silences microRNAs that target key inhibitors of the polarity axis switch. These data support the general belief that EMT-dependent metastasis is a cell-autonomous process. However, our new findings here show that ZEB1 coordinates the components of secretory trafficking machinery, Rab6A and Rab8A, via silencing miR-148a that target these effectors, to drive polarized secretory trafficking toward the leading edge of LUAD cells, to enhance surface exposure of MMP14 so that triggers ECM degradation, accelerates focal adhesion turnover and promotes cancer cell migration. Moreover, the ZEB1-driven secretory trafficking increases the secretions of cytokines and immunomodulation factors, to generate an immunosuppressive tumor microenvironment, and to promote LUAD metastasis. These findings advance a paradigm in which EMT drives LUAD metastasis through a cell non-autonomous mechanism. The novelty rests in our results that demonstrate a transcriptional governance of polarized vesicular transport, providing the potential to target ZEB1-driven secretory pathways for the purpose of blocking metastasis in LUAD. Citation Format: Guan-Yu Xiao, Xiaochao Tan, Leticia Rodriguez, Xin Liu, Jiang Yu, Mayra Vasquez, Hai Tran, William Russell, Don Gibbons, Jonathan Kurie. The EMT activator ZEB1 initiates polarized secretion of pro-tumorigenic effector proteins to drive lung adenocarcinoma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 673.

Sejarah media penyiaran dunia dimulai ketika ahli fisika Jerman bernama Heinrich Hertz pada tahun 1887 berhasil mengirim dan menerima gelombang radio. Upaya Hertz itu kemudian dilanjutkan oleh Guglielmo Marconi (1874-1937) dari Italia yang sukses mengirimkan sinyal morse berupa titik dan garis dari sebuah pemancar kepada suatu alat penerima. Sinyal yang dikirimkan Marconi berhasil menyeberangi Samudra Atlantik pada tahun 1901 dengan menggunakan gelombang elektromagnetik. Stasiun radio pertama muncul ketika seorang tehnik bernama Frank Conrad di Pittsburgh AS, pada tahun 1920 secara iseng-iseng sebagai bagian dari hobi, membangun sebuah pemancar radio di garasi rumahnya. Conrad menyiarkan lagu-lagu, mengumumkan hasil pertandingan olahraga dan menyiarkan instrumen musik yang dimainkan putranya sendiri. Dalam waktu singkat, Conrad berhasil mendapatkan banyak pendengar seiring dengan meningkatnya penjualan pesawat radio ketika itu. Stasiun radio yang dibangun Conrad itu kemudian diberi nama KDKA dan masih tetap mengudara hingga saat ini, menjadikannya sebagai stasiun tertua di Amerika dan mungkin juga didunia. Di Provinsi Aceh perkembangan siaran radio saat ini berkembang dengan pesat mengikuti perubahan zaman dan aturan yang berlaku. Sampai saat ini program-program pada siaran Radio Swasta masih banyak diminati oleh pendengarnya disetiap lapisan masyarakat mulai dari kalangan anak-anak, remaja, hingga dewasa. Program-program siaran Radio Swasta di Aceh khususnya kabupaten Aceh Tamiang banyak menyiarkan siaran-siaran yang berbau Islami hingga siaran-siaran umum dan Pemerintahan, sehingga berbagai program radio menjadi lebih baik dan sesuai. Program Penyiaran tentang Ke-Islaman tersebut disiarkan pada Radio Birama Indah 103,7 FM untuk disiarkan di wilayah Aceh Tamiang guna mendukung terciptanya ketentraman dan kesejahteraan berdasarkan ajaran Islam yang diajarkan oleh baginda Rasulullah SAW sebagai suri tauladan bagi kita semua khususnya umat Islam diseluruh penjuru dunia.

BackgroundOsteoarthritis (OA) is the most common chronic joint disease and is characterized by damage to the articular cartilage. To date, no cure exists (1). Previously, an association between polymorphisms in the acidic leucine-rich nuclear phosphoprotein-32A (ANP32A) gene and OA was described (2). Our group showed that ANP32A plays a key protective role in OA by preventing oxidative stress (3). In OA cartilage, ANP32A is downregulated as compared to non-OA cartilage (3).ObjectivesMaintaining levels of ANP32A seems crucial to preserve joint health, yet the factors that regulate ANP32A in the joint remain unknown. In this study, we aim to investigate factors that regulate ANP32A expression since such knowledge may lead to the identification of specific novel targets for OA therapy.MethodsA bioinformatic analysis pipeline was applied to identify potential regulatory factors of ANP32A expression. We then investigated two selected pathways in vitro, using pharmacological compounds and incubation in a hypoxia chamber in the human articular chondrocyte cell line C28/I2 and in primary human articular chondrocytes (hACs). The effects on ANP32A expression were evaluated using RT-qPCR and Western blot analysis. For in vivo validation, we performed immunohistochemistry to determine ANP32A protein levels in several mouse models.ResultsThe bioinformatic analysis pipeline identified the Wnt signaling pathway and the hypoxia pathway as likely regulatory pathways of ANP32A expression. Wnt hyper-activation is known to contribute to OA (4). We found that Wnt hyper-activation with CHIR99021 decreased ANP32A expression at the mRNA level and at the protein level in hACs. In vivo, ANP32A protein expression was decreased in Frzb knock-out mice and wild-type mice that were treated with CHIR99021 intra-articularly (two Wnt gain-of-function mouse models). ANP32A protein levels increased in wild-type mice upon intra-articular injection of the Wnt inhibitor XAV939. In healthy conditions, cartilage is in hypoxic status but in OA, hypoxia is lost (5). Treatment with the hypoxia mimetic IOX2 and incubation in a hypoxia chamber increased ANP32A expression in vitro in both the C28/I2 cell line and in hACs. In vivo, ANP32A expression was increased in wild-type mice treated with the hypoxia mimetic IOX2 intra-articularly after induction of OA by destabilization of the medial meniscus (DMM).ConclusionLimiting activation of Wnt signaling and maintaining the homeostatic hypoxic environment within the joint are strategies to sustain ANP32A levels and its protective effect against the development of osteoarthritis.References[1]OARSI White Paper - Osteoarthritis as a serious disease 2016 [Available from: https://www.oarsi.org/sites/default/files/docs/2016/oarsi_white_paper_oa-serious-disease.pdf].[2]Valdes AM, Lories RJ, van Meurs JB, Kerkhof H, Doherty S, Hofman A, et al. Variation at the ANP32A gene is associated with risk of hip osteoarthritis in women. Arthritis Rheum. 2009;60(7):2046-54.[3]Cornelis FMF, Monteagudo S, Guns LKA, den Hollander W, Nelissen R, Storms L, et al. ANP32A regulates ATM expression and prevents oxidative stress in cartilage, brain, and bone. Sci Transl Med. 2018;10(458).[4]Lories RJ, Monteagudo S. Review Article: Is Wnt Signaling an Attractive Target for the Treatment of Osteoarthritis? Rheumatol Ther. 2020;7(2):259-70.[5]Bouaziz W, Sigaux J, Modrowski D, Devignes CS, Funck-Brentano T, Richette P, et al. Interaction of HIF1alpha and beta-catenin inhibits matrix metalloproteinase 13 expression and prevents cartilage damage in mice. Proc Natl Acad Sci U S A. 2016;113(19):5453-8.AcknowledgementsFellowship FWO (The Research Foundation – Flanders)Disclosure of InterestsJolien Quintiens: None declared, Frederique M.F. Cornelis: None declared, Astrid De Roover: None declared, Ana Escribano Núñez: None declared, Silvia Monteagudo: None declared, Rik Lories Speakers bureau: consultancy & speaker fees from BioSplice (formerly Samumed), Consultant of: consultancy & speaker fees from BioSplice (formerly Samumed)

BackgroundTumor heterogeneity assessment may help predict response to immunotherapy. In melanoma mouse models, tumor heterogeneity impaired immune response.1 In addition, among lung cancer patients receiving immunotherapy, the high clonal neoantigen group had favorable survival and outcomes.2 Ideal methods of quantifying tumor heterogeneity are multiple biopsies or autopsy. However, these are not feasible in routine clinical practice. Circulating tumor DNA (ctDNA) is emerging as an alternative. Here, we reviewed the current state of tumor heterogeneity quantification from ctDNA. Furthermore, we propose a new tumor heterogeneity index(THI) based on our own scoring system, utilizing both ctDNA and tissue DNA.MethodsSystematic literature search on Pubmed was conducted up to August 18, 2020. A scoring system and THI were theoretically derived.ResultsTwo studies suggested their own methods of assessing tumor heterogeneity. One suggested clustering mutations with Pyclone,3 and the other suggested using the ratio of allele frequency (AF) to the maximum somatic allele frequency (MSAF).4 According to the former, the mutations in the highest cellular prevalence cluster can be defined as clonal mutations. According to the latter, the mutations with AF/MSAF<10% can be defined as subclonal mutations. To date, there have been no studies on utilizing both ctDNA and tissue DNA simultaneously to quantify tumor heterogeneity. We hypothesize that a mutation found in only one of either ctDNA or tissue DNA has a higher chance of being subclonal.We suggest a scoring system based on the previously mentioned methods to estimate the probability for a mutant allele to be subclonal. Adding up the points that correspond to the conditions results in a subclonality score (table 1). In a given ctDNA, the number of alleles with a subclonality score greater than or equal to 2 divided by the total number of alleles is defined as blood THI (bTHI) (figure 1). We can repeat the same calculation in a given tissue DNA for tissue THI (tTHI) (figure 2). Finally, we define composite THI (cTHI) as the mean of bTHI and tTHI.Abstract 18 Table 1Subclonality scoreAbstract 18 Figure 1Hypothetical distribution of all alleles found in ctDNA bTHI = the number of alleles with a subclonality score greater than or equal to 2/the total number of alleles found in ctDNA = 10/20 =50%Abstract 18 Figure 2Hypothetical distribution of all alleles found in tissue DNA tTHI= the number of alleles with a subclonality score greater than or equal to 2/the total number of alleles found in tissue DNA = 16/40 = 40% cTHI= (bTHI + tTHI)/2 = 45%ConclusionsTumor heterogeneity is becoming an important biomarker for predicting response to immunotherapy. Because autopsy and multiple biopsies are not feasible, utilizing both ctDNA and tissue DNA is the most comprehensive and practical approach. Therefore, we propose cTHI, for the first time, as a quantification measure of tumor heterogeneity.ReferencesWolf Y, Bartok O. UVB-Induced Tumor Heterogeneity Diminishes Immune Response in Melanoma. Cell 2019;179:219–235.McGranahan N, Swanton C. Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade. Science 2016;351:1463–1469.Ma F, Guan Y. Assessing tumor heterogeneity using ctDNA to predict and monitor therapeutic response in metastatic breast cancer. Int J Cancer 2020;146:1359–1368.Liu Z, Xie Z. Presence of allele frequency heterogeneity defined by ctDNA profiling predicts unfavorable overall survival of NSCLC. Transl Lung Cancer Res 2019;8:1045–1050.

The pandemic of COVID-19 has afflicted every individual and has initiated a cascade of directly or indirectly involved events in precipitating mental health issues. The human species is a wanderer and hunter-gatherer by nature, and physical social distancing and nationwide lockdown have confined an individual to physical isolation. The present review article was conceived to address psychosocial and other issues and their aetiology related to the current pandemic of COVID-19. The elderly age group has most suffered the wrath of SARS-CoV-2, and social isolation as a preventive measure may further induce mental health issues. Animal model studies have demonstrated an inappropriate interacting endogenous neurotransmitter milieu of dopamine, serotonin, glutamate, and opioids, induced by social isolation that could probably lead to observable phenomena of deviant psychosocial behavior. Conflicting and manipulated information related to COVID-19 on social media has also been recognized as a global threat. Psychological stress during the current pandemic in frontline health care workers, migrant workers, children, and adolescents is also a serious concern. Mental health issues in the current situation could also be induced by being quarantined, uncertainty in business, jobs, economy, hampered academic activities, increased screen time on social media, and domestic violence incidences. The gravity of mental health issues associated with the pandemic of COVID-19 should be identified at the earliest. Mental health organization dedicated to current and future pandemics should be established along with Government policies addressing psychological issues to prevent and treat mental health issues need to be developed. References World Health Organization (WHO) Coronavirus Disease (COVID-19) Dashboard. 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Abstract Background Traumatic brain injury, one of the leading causes of death in adults under 40 years of age in the world, is frequently caused by mechanical shock, resulting in diffuse neuronal damage and long-term cognitive dysfunction. Many existing TBI animal models revival with expensive equipment or special room are needed or the processes of operations are complex and not easy to be widely used. Therefore, a simpler TBI model needs to be designed. Methods Our TBI model is an innovation of the modeling method through air guns shutting rubber bullets. A core facet is the application of our designed rubber bullet impact device. It could focus the hitting power to the fixed site of the brain, thus triggering a mild closed head injury. Moreover, the degree of damage can be adjusted by the times of shots. Results Our model induced blood-brain barrier leakage and diffused neuronal damage. Besides, it led to an increased level of Tau phosphorylation and resulted in cognitive dysfunction within several weeks post-injury. Conclusion Our TBI model is not only simple and time-saving but also can simulate mild brain injuries in clinical. It is suitable for exploring pathobiological mechanisms as well as a screening of potential therapies for TBI.

Abstract As the performance of current fall risk assessment tools is limited, clinicians face significant challenges in identifying patients at risk of falling. This study proposes an automatic fall risk prediction model based on eXtreme gradient boosting (XGB), using a data-driven approach to the standardized medical records. This study analyzed a cohort of 639 participants (297 fall patients and 342 controls) from Chang Gung Memorial Hospital, Chiayi Branch, Taiwan. A derivation cohort of 507 participants (257 fall patients and 250 controls) was collected for constructing the prediction model using the XGB algorithm. A comparative validation of XGB and the Morse Fall Scale (MFS) was conducted with a prospective cohort of 132 participants (40 fall patients and 92 controls). The areas under the curves (AUCs) of the receiver operating characteristic (ROC) curves were used to compare the prediction models. This machine learning method provided a higher sensitivity than the standard method for fall risk stratification. In addition, the most important predictors found (Department of Neuro-Rehabilitation, Department of Surgery, cardiovascular medication use, admission from the Emergency Department, and bed rest) provided new information on in-hospital fall event prediction and the identification of patients with a high fall risk.

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IntroductionI stumble to my feet breathing heavily and, over the roar of a tank, a nearby soldier yells right into my face: “We’re surrounded! We have to hold this line!” I follow him, moving past burning debris and wounded men being helped walk back in the opposite direction. Shells explode around me, a whistle sounds, and then the Hun attack; shadowy figures that I fire upon as they approach through the battlefield fog and smoke. I shoot some. I take cover behind walls as others fire back. I reload the weapon. I am hit by incoming fire, and a red damage indicator appears onscreen, so I move to a better cover position. As I am hit again and again, the image becomes blurry and appears as if in slow-motion, the sound also becoming muffled. As an enemy wielding a flame-thrower appears and blasts me with thick fire, my avatar gasps and collapses. The screen fades to black.So far, so very normal in the World War One themed first-person shooter Battlefield 1 (Electronic Arts 2016). But then the game does something unanticipated. I expect to reappear—or respawn—in the same scenario to play better, to stay in the fight longer. Instead, the camera view switches to an external position, craning upwards cinematically from my character’s dying body. Text superimposed over the view indicates the minimalist epitaph: “Harvey Nottoway 1889-1918.” The camera view then races backwards, high over the battlefield and finally settles into position behind a mounted machine-gun further back from the frontline as the enemy advances closer. Immediately I commence shooting, mowing down German troops as they enter our trenches. Soon I am hit and knocked away from the machine-gun. Picking up a shotgun I start shooting the enemy at close-quarters, until I am once again overrun and my character collapses. Now the onscreen text states I was playing as “Dean Stevenson 1899-1918.”I have attempted this prologue to the Battlefield 1 campaign a number of times. No matter how skilfully I play, or how effectively I simply run away and hide from the combat, this pattern continues: the structure of the game forces the player’s avatar to be repeatedly killed in order for the narrative to progress. Over a series of player deaths, respawning as an entirely new character each time, the combat grows in ferocity and the music also becomes increasingly frenetic. The fighting turns to hand-to-hand combat, or shovel-to-head combat to be more precise, and eventually an artillery barrage wipes everybody out (Figure 1). At this point, the prologue is complete and the gamer may continue in a variety of single-player episodes in different theatres of WW1, each of which is structured according to the normal rules of combat games: when your avatar is killed, you respawn at the most recent checkpoint for a follow-up attempt.What are we to make of this alternative narrative structure deployed by the opening episode of Battlefield 1? In contrast to the normal video-game affordances of re-playability until completion, this narrative necessitation of death is in some ways motivated by the onscreen text that introduces the prologue: “What follows is frontline combat. You are not expected to survive.” Certainly it is true that the rest of the game (either single-player or in its online multiplayer deathmatch mode) follows the predictable pattern of dying, replaying, completing. And also we would not expect Battlefield 1 to be motivated primarily by a kind of historical fidelity given that an earlier instalment in the series, Battlefield 1942 (2002) was described by one reviewer as:a comic book version of WWII. The fact that any player can casually hop into a tank, drive around, hop out and pick off an enemy soldier with a sniper rifle, hop into a plane, parachute out, and then call in artillery fire (within the span of a few minutes) should tell you a lot about the game. (Osborne)However what is happening in this will-to-die structure of the game’s prologue represents an alternative and affectively unsettling game experience both in its ludological structure as well as its affective impact. Defamiliarization and Humanization Drawing upon a phenomenology of game-play, whereby the scholar examines the game “as played” (see Atkins and Kryzwinska; Keogh; Wilson) to consider how the text reveals itself to the player, I argue that the introductory single-player episode of Battlefield 1 functions to create a defamiliarizing effect on the player. Defamiliarization, the Russian Formalist term for the effect created by art when some unusual aspect of a text challenges accepted perceptions and/or representations (Schklovski; Thompson), is a remarkably common effect created by the techniques used in combat cinema and video-games. This is unsurprising. After all, warfare is one of the very examples Schklovski uses as something that audiences have developed habituated responses to and which artworks must defamiliarize. The effect may be created by many techniques in a text, and in certain cases a work may defamiliarize even its own form. For instance, recent work on the violence in Saving Private Ryan shows that during the lengthy Omaha Beach sequence, the most vivid instances of violence—including the famous shot of a soldier picking up his dismembered arm—occur well after the audience has potentially become inured to the onslaught of the earlier frequent, but less graphic, carnage (Bender Film Style and WW2). To make these moments stand out with equivalent horrific impact against the background of the Normandy beach bloodbath Spielberg also treats them with a stuttered frame effect and accompanying audio distortion, motivated (to use a related Formalist term) by the character’s apparent concussion and temporary disorientation. Effectively a sequence of point of view shots then, this moment in Private Ryan has become a model for many other war texts, and indeed the player’s death in the opening sequence of Battlefield 1 is portrayed using a very similar (though not identical) audio-visual treatment (Figure 2).Although the Formalists never played videogames, recent scholarship has approached the medium from a similar perspective. For example, Brendan Keogh has focused on the challenges to traditional videogame pleasure generated by the 2012 dystopian shooter Spec Ops: The Line. Keogh notes that the game developers intended to create displeasure and “[forcing] the player to consider what is obscured in the pixilation of war” by, for instance, having them kill fellow American troops in order for the game narrative to continue (Keogh 9). In addition, the game openly taunts the player’s expectations of entertainment based, uncritical run-and-gun gameplay with onscreen text during level loading periods such as “Do you feel like a hero yet?” (8).These kinds of challenges to the expectations of entertainment in combat shooters are found also in one sequence from the 2009 game Call of Duty: Modern Warfare 2 in which the player—as an undercover operative—is forced to participate in a terrorist attack in which civilians are killed (Figure 3). While playing that level, titled “No Russian,” Timothy Welsh argues: “The player may shoot the unarmed civilians or not; the level still creeps slowly forward regardless” (Welsh 409). In Welsh’s analysis, this level emerges as an unusual attempt by a popular video game to “humanize” the non-playing characters that are ordinarily gunned down without any critical and self-reflective thought by the player in most shooter games. The player is forced into a scenario in which they must make a highly difficult ethical choice, but the game will show civilians being killed either way.In contrast to the usual criticisms of violent video games—eg., that they may be held responsible for school shootings, increased adolescent aggression and so on —the “No Russian” sequence drew dramatic complaints of being a “terrorist simulator” (Welsh 389). But for Welsh this ethical choice facing the player, to shoot or not to shoot civilians, raises the game to a textual experience offering self-inspection. As in the fictional theme park of Westworld (HBO 2016), it does not really matter to the digital victim if a player kills them, but it should—and does—matter to the player. There are no external consequences to killing a computer game character composed only of pixels, or killing/raping a robot in the Westworld theme park, however there are internal consequences: it makes you a killer, or a rapist (see Harris and Bloom).Thus, from the perspective of defamiliarization, the game can be regarded as creating the effect that Matthew Payne has labelled “critical displeasure.” Writing about the way this is created by Spec Ops, Payne argues that:the result is a game that wields its affective distance as a critique of the necessary illusion that all military shooters trade in, but one that so few acknowledge. In particular, the game’s brutal mise-en-scène, its intertextual references to other war media, and its real and imagined opportunities for player choice, create a discordant feeling that lays bare the ease with which most video war games indulge in their power fantasies. (Payne 270)There is then, a minor tradition of alternative military-themed video game works that attempt to invite or enable the player to conduct a kind of ethical self-examination around their engagement with interactive representations of war via particular incursions of realism. The critical displeasure invoked by texts such as Spec Ops and the “No Russian” level of Call of Duty is particularly interesting in light of another military game that was ultimately cancelled by the publisher after it received public criticism. Titled Six Days in Fallujah, the game was developed with the participation of Marines who had fought in that real life battle and aimed to depict the events as they unfolded in 2004 during the campaign in Iraq. As Justin Rashid argues:the controversy that arose around Six Days in Fallujah was, of course, a result of the view that commercial video games can only ever be pure entertainment; games do not have the authority or credibility to be part of a serious debate. (Rashid 17)On this basis, perhaps a criterial attribute of an acceptable alternative military game is that there is enough familiarity to evoke some critical distance, but not too much familiarity that the player must think about legitimately real-life consequences and impact. After all, Call of Duty was a successful release, even amid the controversy of “No Russian.” This makes sense as the level does not really challenge the overall enjoyment of the game. The novelty of the level, on the one hand, is that it is merely one part of the general narrative and cannot be regarded as representative of the whole game experience. On the other hand, because none of the events and scenarios have a clear indexical relationship to real-world terrorist attacks (at least prior to the Brussels attack in 2016) it is easy to play the ethical choice of shooting or not shooting civilians as a mental exercise rather than a reflection on something that really happened. This is the same lesson learned by the developers of the 2010 game Medal of Honor who ultimately changed the name of the enemy soldiers from “The Taliban” to “OPFOR” (standing in for a generic “Opposing Forces”) after facing pressure from the US and UK Military who claimed that the multiplayer capacities of the game enabled players to play as the Taliban (see Rashid). Conclusion: Affective Friction in Battlefield 1In important ways then, these game experiences are precursors to Battlefield 1’s single player prologue. However, the latter does not attempt a wholesale deconstruction of the genre—as does Spec Ops—or represent an attempt to humanise (or perhaps re-humanise) the non-playable victim characters as Welsh suggests “No Russian” attempts to do. Battlefield 1’s opening structure of death-and-respawn-as-different-character can be read as humanizing the player’s avatar. But most importantly, I take Battlefield’s initially unusual gameplay as an aesthetic attempt to set a particular tone to the game. Motivated by the general cultural attitude of deferential respect for the Great War, Battlefield 1 takes an almost austere stance toward the violence depicted, paradoxically even as this impact is muted in the later gameplay structured according to normal multiplayer deathmatch rules of run-and-gun killing. The futility implied by the player’s constant dying is clearly motivated by an attempt at realism as one of the cultural memories of World War One is the sheer likelihood of being killed, whether as a frontline soldier or a citizen of a country engaged in combat (see Kramer). For Battlefield 1, the repeated dying is really part of the text’s aesthetic engagement. For this reason I prefer the term affective friction rather than critical displeasure. The austere tone of the game is indicated early, just prior to the prologue gameplay with onscreen text that reads:Battlefield 1 is based on events that unfolded over 100 years agoMore than 60 million soldiers fought in “The War to End All Wars”It ended nothing.Yet it changed the world forever. At a simple level, the player’s experience of being killed in order for the next part of the narrative to progress evokes this sense of futility. There have been real responses indicating this, for instance one reviewer argues that the structure is “a powerful treatment” (Howley). But there is potential for increased engagement with the game itself as the structure breaks the replay-cycle of usual games. For instance, another reviewer responds to the overall single-player campaign by suggesting “It is not something you can sit down and play through and not experience on a higher level than just clicking a mouse and tapping a keyboard” (Simpson). This affective friction amplifies, and draws attention to, the other advances in violent stylistics presented in the game. For instance, although the standard onscreen visual distortions are used to show character damage and the direction from which the attack came, the game does use slow-motion to draw out the character’s death. In addition, the game features incidental battlefield details of shell-shock, such as soldiers simply holding the head in their hands, frozen as the battle rages around them (Figure 4). The presence of flame-thrower troops, and subsequently the depictions of characters running as they burn to death are also significant developments in violent aesthetics from earlier games. These elements of violence are constitutive of the affective friction. We may marvel at the technical achievement of such real-time rendering of dynamic fire and the artistic care given to animate deaths and shell-shock depictions. But simultaneously, these “violent delights”—to borrow from Westworld’s citation of Shakespeare—are innovations upon the depictions of earlier games, even contemporary, combat games. Indeed, one critic has almost ashamedly noted: “For a game about one of the most horrific wars in human history, it sure is pretty” (Kain).These violent depictions show a continuation in the tradition of increased detail which has been linked to a model of “reported realism” as a means of understanding audience’s claims of realism in combat films and modern videogames as a result primarily of their hypersaturated audio-visual texture (Bender "Blood Splats"). Here, saturation refers not to the specific technical quality of colour saturation but to the densely layered audio-visual structure often found in contemporary films and videogames. For example, thick mixing of soundtracks, details of gore, and nuanced movements (particularly of dying characters) all contribute to a hypersaturated aesthetic which tends to prompt audiences to make claims of realism for a combat text regardless of whether or not these viewers/players have any real world referent for comparison. Of course, there are likely to be players who will simply blast through any shooter game, giving no regard to the critical displeasure offered by Spec Ops narrative choices or the ethical dilemma of “No Russian.” There are also likely to be players who bypass the single-player campaign altogether and only bother with the multiplayer deathmatch experience, which functions in the same way as it does in other shooter games, including the previous Battlefield games. But perhaps the value of this game’s attempt at alternative storytelling, with its emphasis on tone and affect, is that even the “kill-em-all” player may experience a momentary impact from the violence depicted. This is particularly important given that, to borrow from Stephanie Fisher’s argument in regard to WW2 games, many young people encounter the history of warfare through such popular videogames (Fisher). In the centenary period of World War One, especially in Australia amid the present Anzac commemorative moment, the opportunity for young audiences to engage with the significance of the events. As a side-note, the later part of the single-player campaign even has a Gallipoli sequence, though the narrative of this component is designed as an action-hero adventure. Indeed, this is one example of how the alternative dying-to-continue structure of the prologue creates an affective friction against the normal gameplay and narratives that feature in the rest of the text. The ambivalent ways in which this unsettling opening scenario impacts on the remainder of the game-play, including for instance its depiction of PTSD, is illustrated by some industry reviewers. As one reviewer argues, the game does generate the feeling that “war isn’t fun — except when it is” (Plante). From this view, the cognitive challenge created by the will to die in the prologue creates an affective friction with the normalised entertainment inherent in the game’s multiplayer run-and-gun components that dominate the rest of Battlefield 1’s experience. Therefore, although Battlefield 1 ultimately proves to be an entertainment-oriented combat shooter, it is significant that the developers of this major commercial production decided to include an experimental structure to the prologue as a way of generating tone and affect in a fresh way. ReferencesAtkins, Barry, and Tanya Kryzwinska. "Introduction: Videogame, Player, Text." Videogame, Player, Text. Eds. Atkins, Barry and Tanya Kryzwinska. Manchester: Manchester University Press, 2007.Bender, Stuart Marshall. "Blood Splats and Bodily Collapse: Reported Realism and the Perception of Violence in Combat Films and Videogames." Projections 8.2 (2014): 1-25.Bender, Stuart Marshall. Film Style and the World War II Combat Film. 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