bluuebunny/crossref_metadata_2025 huggingface dataset:
Agenda para regularizar las crypto divisas en Costa Rica y sus posibilidades ante el mercado financiero Costarricense
Authors: Cordero Céspedes Alexander , Cordero Ulate Andrés
Published: August 2023

En la presente tesis se desarrolló el tema de la posible agenda para regularizar las Cryto Divisas en Costa Rica y sus posibilidades ante el Mercado Financiero Costarricense, en el año 2023, para el cual, se procedió a describir el problema, su importancia y objetivos específicos; así como los aspectos de antecedentes del problema, para poder delimitar el desarrollo de la posible ruta de la implementación de una regularización de las Cryto divisas  en contexto del presente  trabajo de investigación.
Strategi Komunikasi Publik Humas Pemerintah Kabupaten Sidoarjo dalam Meningkatkan Reputasi Di Masyarakat
Authors: Ismojo Herdono  , Dzaki Makarim
Published: August 2023

Bupati Sidoarjo, Muhdlor Ali, tentu agar reputasi Kabupaten Sidoarjo dipandang baik oleh masyarakat maka membutuhkan peranan humas. Humas memiliki peran antara lain sebagai jembatan atau penghubung antara pihak internal dengan eksternal. Penelitian ini bertujuan untuk mengetahui strategi komunikasi humas Pemerintah Kabupaten Sidoarjo dalam meningkatkan reputasi di masyarakat. Teori yang digunakan adalah strategi komunikasi, komunikasi publik, humas, dan reputasi. Menurut Rogers sebagaimana yang dikutip oleh Hafied Cangara, strategi komunikasi dapat diartikan sebagai suatu rencana yang dibangun dengan tujuan untuk mengubah perilaku manusia dalam jumlah yang besar melalui perpindahan gagasan-gagasan yang baru. Komunikasi publik dapat diartikan sebagai saling bertukar informasi atau pesan dengan beberapa orang yang berada dalam maupun di luar sebuah organisasi, baik secara langsung atau tidak langsung. Metode yang digunakan dalam penelitian ini yaitu metode penelitian kuasi-kualitatif, yakni pendekatan yang tidak murni kualitatif atau belum kualitatif secara hakikat dikarenakan secara bentuk pendekatan ini masih terpengaruhi oleh kebiasaan-kebiasaan pendekatan kuantitatif. Teknik pengumpulan data serta metode penelitian dalam penelitian ini dibagi menjadi dua, yakni menyangkut wawancara, dan data-data dari dokumentasi. Sementara keabsahan data dilakukan melalui triangulasi sumber. Hasil penelitian ini menampakkan strategi komunikasi Humas Pemkab Sidoarjo ialah melalui komunikasi eksternal, internal, vertikal, horizontal, dan diagonal. Kegiatan-kegiatan yang dilakukan oleh Humas Pemkab Sidoarjo melalui pemanfaaatan dan penggunaan media penyebaran informasi ke publik, kualitas sumber daya manusia, dan pembentukan reputasi yang kurang maksimal. Guna merealisasikan tujuan Humas Pemkab Sidoarjo dalam meningkatkan reputasi di masyarakat maka dibutuhkan keaktifan pemanfaatan media baik massa maupun sosial, perbaikan kualitas SDM, dan koordinasi antar dinas.
All taxa biodiversity inventory of the Bois de Bouis estate (Var, France): a 10-year public-private partnership
Authors: Lacoeuilhe Aurélie , Percevault Louise , Ichter Jean , Gourdain Philippe , Herard Katia , Michaud Henri , Poncet Laurent , Ramage Thibault , Roquinarc'h Océane , Withers Phil
Published: August 2023

This data paper describes the results of a 10-year scientific investigation of a biodiversity-rich private golf estate in south-eastern France in partnership with PatriNat (Office français de la biodiversité/Centre national de la recherche scientifique/Muséum national d'Histoire naturelle, Institut de Recherche pour le Développement). In total, 3,160 species and subspecies, including 1,796 arthropods and 1,049 flora, were inventoried and 65 habitat types were surveyed and mapped. This project is the first All taxa biodiversity inventory (ATBI) in a private property in France with all information available in open data.

The 20 datasets of fauna, flora, lichens and habitat types from the Bois de Bouis estate are now publicly available. Between 2012 and 2022, more than 22,000 occurrences were recorded, checked and published in the INPN information system. All this information is available in open access in the French portal OpenObs, operated by PatriNat and in the Global Biodiversity Information Facility (GBIF). This data paper provides an overview of the project, its main results and its contribution to the French National Inventory of Natural Heritage (INPN).

This data paper presents a list eight species never previously recorded to France; three Hymenoptera: Charitopes leucobasis Townes, 1983 (Ichneumonidae), Dryinus tussaci Olmi, 1989 (Dryinidae) and Sparasion munitus Kozlov & Kononova, 1990 (Sparasionidae) and five Diptera: Clusiodes apicalis (Zetterstedt, 1848) (Clusiidae), Dicraeus vagans (Meigen, 1838) (Chloropidae), Stilpon intermedius Raffone, 1994, Stilpon subnubilus Chvala, 1988 and Tachydromia undulata (Strobl, 1906) (Hybotidae).

It also includes a table comparing the project to 18 All-taxa biodiversity inventories in France and Belgium and published for the first time.
The Effect of Timing Between Traumatic Flexor Tendon Injury of the Hand and Surgical Intervention on Repair Failure Rates
Authors: Reddy Rajiv P. , Gao Timothy , Chen Joseph , Goitz Robert , Kaufmann Robert A. , Fowler John R.
Published: August 2023

Flexor tendon injuries of the hand, especially in zone II, pose a challenge for hand surgeons because of the region's intricate pulley mechanism and local avascularity, and post-surgical complications such as repair failure are not uncommon. One proposed predictor of outcomes following flexor tendon repair has been timing of surgery from initial injury. However, the effect of the timing of flexor tendon repair on failure rates remains controversial and understudied. The purpose of this study was to compare the failure rates of zone II flexor tendon repairs in patients at various time intervals from onset of injury. A retrospective chart review was conducted using data from hand surgery specialists at our level 1 trauma center from January 1, 2010, through May 31, 2020. This retrospective review included 407 zone II flexor tendon repairs. The primary outcome was failure of repair. Among 407 flexor tendon repairs, there were 12 reported repair failures. The failure rate was 2.9%. In the non-failure group, the mean number of days between the date of injury and the date of surgery was 7±13 days. For the failure group, this value was 14±17 days. Repairs occurring within 14 days had a failure rate of 2.3%, while repairs occurring beyond 14 days had a failure rate of 7.7%. This study demonstrates that there is a benefit to repairing the tendon within a 14-day window, as evidenced by a lower failure rate. More research is required to determine if other complications and overall health of the hand are also improved when a repair is performed in a more expedient manner. [
Orthopedics
. 2024;47(2):113–117.]
Evaluation and Management of Blunt Head Trauma in the Pediatric Clinic
Authors: Diaz Kane Molly M.
Published: August 2023

Head injuries, and specifically blunt head trauma, are common among pediatric patients of all ages. Patients may present to their primary care provider, to urgent care, or to the emergency department after head trauma. Such injuries may occur as a result of a variety of mechanisms, including falls, motor vehicle collisions, or sports injuries. Clinical decision rules exist to help guide the clinician in the initial evaluation of head injury and in determining when head imaging may be indicated. One such guideline that is widely used in the United States is known as the PECARN (Pediatric Emergency Care Applied Research Network) criteria. Pediatricians should also evaluate for the presence of symptoms consistent with concussion that may occur as a result of blunt head trauma and be familiar with the management and sequelae of concussion and head injuries.

[
Pediatr Ann
. 2023;52(8):e279–e281.]
Gradient estimates in generalized Orlicz spaces for quasilinear elliptic equations via extrapolation
Authors: Wu Ruimin , Jiang Yinsheng , Wang Liyuan
Published: August 2023

The gradient estimates in the generalized Orlicz space for weak solutions of a class of quasi-linear elliptic boundary value problems are obtained using the modern technique of extrapolation. The coefficients are assumed to have small BMO seminorms, and the boundary of the domain is sufficiently flat in the sense of Reifenberg. As a corollary, we apply our results to the variable Lebesgue spaces.
Virus: Neoliberalni detektiv v imuni slovenski družbi
Authors: Mlačnik Primož
Published: August 2023

Članek s pomočjo imunitarne paradigme Jacquesa Derridaja in Roberta Esposita analizira slovenski detektivski roman Virus. V prvem delu raziskujemo vezi med neoliberalizmom in vzponom slovenske avtoritarne države v času prvega vala pandemije covid-19. V drugem delu pokažemo, da je neoliberalni etos reprezentiran v liku sebičnega in samodiscipliniranega detektiva, v značaju nedružbenega in privatiziranega zločina ter v značaju samopravičniškega zločinca. Dejanja detektiva in zločinca so avtoimunitarna. Čeprav protislovno oba delujeta kot zavetnika skupnega dobrega, oba sledita zasebnim interesom. V zadnjem delu dekonstruiramo binarna nasprotja, na katerih temelji dvojni status umorjenih žrtev, ki so zločinsko odgovorne in posedujejo nenavadno (neheteronormativno) ali patološko spolno identiteto.
Gverilska multituda
Authors: Liponik Vesna
Published: August 2023

Gverilska multituda
Foucault’s Adventure in Iran and His Last “Turn”
Authors: Reihanzadeh Arsalan
Published: August 2023

In light of the Iranian uprising of 1978–79, Foucault preferred to use the term revolt instead of revolution. In the first part of the article, we attempt to go beyond the limits of the nonspecific distinction he made between revolt and revolution by drawing on Furio Jesi’s phenomenology of revolt and showing in what sense the Iranian uprising can be considered a revolt. In the second part, the article highlights the connection between the Iranian uprising and Foucault’s later works by arguing that his adventure in Iran effectively shaped his final intellectual trajectory. By examining the tension between the transcendental and the historical in Foucault’s works, we propose to read his Iranian experience against the backdrop of such tension to better understand his different approach to the question of the transcendental in his final years.
Mathematical Science of Being
Authors: Germek Magdalena
Published: August 2023

In the present article, we have demonstrated that it is important to understand the equating of mathematics with ontology in Badiou’s philosophy, taking into account the necessary connection between rational materialism and ontological realism. Only in this way can we truly understand Badiou’s fundamental thesis that thinking and being are the same. Philosophy is not ontology and it is not a true procedure, but a thought that arises by being conditioned with the generic thoughts of all four truth procedures (art, politics, science, and love), which in turn does not mean that it cannot speak in the name of truth and ontology.
World at the Border: The Cosmopolitan Ideal between Loss and Multiplication
Authors: Benčin Rok
Published: August 2023

The article examines the transformations of the philosophical concept of world as it appears in the cosmopolitan tradition of political thought and its relation to the problem of the border. It focuses particularly on how world is understood as either lost or multiplied in the contexts of modernity, globalisation, and migration. The article discusses postcolonial conceptions of cosmopolitics and the political philosophy of Hannah Arendt to show how the universal ideal of the world is replaced by singular constructions of worlds in terms of the experience of migrants and refugees or the phenomenological horizon of political action. I conclude by suggesting that Jacques Rancière’s understanding of politics as a conflict of worlds can take us beyond the traps of both cosmopolitan universalism and the phenomenological singularity of being-in-the-world.
Disorientation in a Time of the Absence of Limits
Authors: Šumič Riha Jelica
Published: August 2023

Seen from the perspective of the inconsistency of the Other, the post-truth era can be considered to be an era emerging from a crisis in belief in the existence of the Other, which is to be taken in a twofold sense: as a belief in the Other of the Other, that is, the Other of Law, and a belief in the Other considered as the subject supposed to know. Insofar as the contemporary subject does not want to know anything about this “condition of belief” without which no knowledge, and therefore no truth, are possible, the crisis of belief affect both, the Other and the subject. This can be seen in the fact that the failing belief in the Other and knowledge, considered as a distinctive feature of our profoundly unbelieving times, is accompanied by an unprecedented rise in anxiety at the social level, as contemporary subjects who do not believe in the (existence of the) Other are singularly defenceless before the irruption of the real. With truth losing its power to name the real, the subject itself as a singular response to the real is becoming ever more precarious. Which is why, when faced with the erratic irruption of the real, contemporary subjects are condemned to a desperate search for certainties.
Artificial Intelligence for Information Security
Authors: Isaoglu Lubana , Yiltas-Kaplan Derya
Published: August 2023

The internet is becoming a widely used source of both online services and information. The primary function of the internet is to move data between nodes. It is a vast collection of interconnected devices and computers, and it has become the target of different cybercrimes. Due to the rapid development of the internet and the increasing number of cyber-attacks, the detection of these attacks has become more important. The traditional security methods are not enough to face these threats; thus, researchers have been developing smarter techniques such as deep learning and machine learning to address these problems. This chapter explores the various aspects of deep learning and machine learning on information security. It also focuses on the challenges and motivations of this new technology. This chapter reviewed previously conducted surveys and papers in the last two years in the field; thus, it can be used by researchers who are interested in learning more about the various aspects and trends of deep learning and machine learning in information security.
Zero Day Vulnerabilities Assessments, Exploits Detection, and Various Design Patterns in Cyber Software
Authors: Thapa Vidhanth Maan , Srivastava Sudhanshu , Garg Shelly
Published: August 2023

In this technology-driven era, software development and maintenance is a rapidly growing domain and is predestined to thrive over the coming decade. But the growing demand for software solutions also brings its own implications. Software vulnerabilities are the most crucial of these. Software Vulnerabilities can be referred to as weaknesses or shortcomings of the software solutions which increase the risks of exploitation of resources and information. In the past few years, the number of exploits has been increasing rapidly, reaching an all-time high in 2021 affecting more than 100 million people worldwide. Although, even with the presence of existing vulnerability management models and highly secure tools and frameworks, software vulnerabilities are harder to identify and resolve as they may not be independent, and resolving them may cause other vulnerabilities. Moreover, a majority of the exploit are caused due to known vulnerabilities and zero-day vulnerabilities..
Anti-Money Laundering Regulations and Entrepreneurship
Authors: Ajide Folorunsho
Published: August 2023

The implications of money laundering activities in developing economies have yielded mixed findings. However, existing studies have not specifically examined the role of anti-money laundering (AML) regulations in relation to entrepreneurial development. This study aims to fill this gap by analyzing the impact of AML regulations on entrepreneurship in Africa. The analysis utilizes data from 21 African nations for 2012-2018. The empirical analysis employs panel-corrected standard errors (PCSEs) and instrumental variable (IV) regression. The results demonstrate a significant positive impact of AML regulations on entrepreneurship. It is also revealed that AML regulations bolster institutional quality, thereby fostering entrepreneurship. Moreover, the study finds that AML measures facilitate financial development, further promoting entrepreneurial growth in Africa. Importantly, these results remain robust even after conducting a thorough robustness check. Consequently, the study advocates for the implementation of effective AML regulations to enhance the African business environment.
Corruption, Government Effectiveness, and Financial Development
Authors: Alshubiri Faris Nasif
Published: August 2023

Government effectiveness has emerged as an important factor in the relationship between corruption and the process of financial development. This chapter explores the relationship between corruption control, government effectiveness and financial development in 14 low- and high-income countries over the period 2005–2020 to determine the applicability of the grease the wheels and the sand the wheels hypotheses. Panel data for the two country groups were analysed after overcoming heterogeneity and endogeneity issues. Diagnostic tests were used to increase robustness. The analysis revealed that an increase in corruption control led to a decrease in financial development due to highly effective governments in high-income countries. In contrast, an increase in corruption control led to an increase in financial development due to inefficient governments in low-income countries. The main conclusions of the study are that the grease the wheels hypothesis is applicable to high-income countries and the sand the wheels one is applicable to low-income countries.
Inconsistencies in the Disclosures of Discount Rates
Authors: Assunção Miguel , Albuquerque Fábio
Published: August 2023

The use of the discount rate within financial reporting deserves particular attention from international standard-setting bodies because it has achieved prominence over time. This chapter uses archival research as a method and content analysis as a technique to assess mandatory and voluntary disclosure levels related to discount rates. The consolidated accounts for the year 2020 of listed entities in Euronext Lisbon are the sources of information. The results show differences in mandatory and disclosure levels regarding IAS/IFRS under analysis. Disclosures related to discount rates have been assessed either as an incidental topic or from a limited-scope analysis. Therefore, understanding the proper set of items to be disclosed and the factors that explain its disclosure, aligned with the efforts to ensure its compliance by legal authorities, may reduce asymmetries and enhance transparency in entities' financial reporting.
Occupational Fraud in the Highly Regulated Banking Industry
Authors: Beemamol M.
Published: August 2023

Banking has witnessed a significant surge in occupational fraud globally, establishing it as the most impacted industry. Similarly, the banking industry in India has also experienced a rise in fraudulent activities perpetrated by bank officials. This chapter aims to investigate five notable cases of high-value occupational fraud committed by top management personnel in Indian banks. Multiple contributing factors have been identified, including greed, social comparison, corrupt ideology, lack of ethics and morality, a sense of entitlement and ego, hubris, and a hyper-competitive culture. These factors played a pivotal role in enticing top management to exploit fraud opportunities and collaborate with other bank employees and corporate borrowers for personal gain. The prevalence of collusive fraud within the banking sector is an alarming trend that jeopardizes the industry's integrity, potentially foreshadowing its decline. The findings of this study establish a robust foundation for future empirical research and provide valuable insights to enrich the existing body of literature.
Corporate Climate Change-Related Accounting and Taxation
Authors: Monteiro Sónia , Lemos Kátia , Martin Jose Jurado , Martín-Zamora María-Pilar
Published: August 2023

This chapter aims to provide an overview of corporate climate change-related accounting and taxation practices in Portugal and Spain. The Portuguese Accounting and Financial Reporting Standard No. 26: Environmental Matters and the Spanish Royal Decree 602/2016 establish the current accounting treatment for greenhouse (GHG) emissions. While the initial measurement basis is the same in both countries (cost when licenses are acquired or fair value when obtained for free), the accounting regulations prescribe different accounting treatments for the initial recognition. In Spain, allowances are recognized as inventories, while in Portugal, they are recognized as intangible assets. Interestingly, the Portuguese regulation uses the FIFO inventory method to calculate the amortization of the intangible asset. Regarding corporate income tax, there is no specific treatment for emission allowances in either country's tax legislation. Additionally, a reverse-charge mechanism on CO2 emissions trading has been introduced in Europe, resulting in no differences between countries concerning VAT.
Green Taxation for SMEs' Digital Transformation of Accounting
Authors: Lemos Kátia , Dinis Ana , Serra Sara
Published: August 2023

The dematerialization of documents and the resulting contribution to greener and more sustainable practices should be on the agenda of businesses. Companies, especially SMEs, need to be particularly aware of the relevance of digitalization for environmental sustainability, and the role of governments in promoting the digitalization of companies is very important for its success. The present research focuses on the study of tax incentives for the digital transformation of accounting granted to Portuguese micro and SMEs, specifically for the implementation of SAF-T (PT) for accounting, the QR Code®, and the ATCUD. An exploratory data analysis methodology was adopted, considering all Portuguese micro and SMEs that participated in the tax incentive for digital transformation in 2020 and 2021. The authors conclude that the tax incentives studied have not been a practice adopted by Portuguese micro and SMEs, which may reveal a lack of disclosure by the government and knowledge by companies.
Tourist Taxes and Sustainability
Authors: Miranda Eva Maria , Gonçalves Soraia , Vareiro Laurentina
Published: August 2023

As the global tourism industry continues to grow, concerns about its impact on the environment and local communities have increased. In response to these concerns, many destinations have implemented tourist taxes as a policy tool to manage the negative impacts of tourism on destinations and promote sustainable tourism. This chapter presents a systematic literature review on the relationship between tourist taxes and sustainability, using the Bibliometrix R software and a content analysis. The aim of this review is to provide a comprehensive analysis of the existing research, identifying gaps in the literature and suggesting areas for future research. The results demonstrate two main categories, concerning demand and supply dimensions. The issue of sustainability is transversal to the articles analyzed, highlighting its evolution in a holistic way and encompassing the economic, social, and environmental pillars.
Testing of a novel liposomal anti-Pseudomonas vaccine based on outer-membrane protein F (OprF) of Pseudomonas aeruginosa
Authors: Dissanayake Ravindika , Nada Shadia , Thanvi Radhika , Sebilleau Chloe O , Prestwich Erin G , Sucheck Steven J , Wall Katherine A
Published: August 2023

Pseudomonas aeruginosa is an opportunistic pathogen associated with an increased risk of morbidity and mortality among cystic fibrosis patients and immunocompromised individuals. Current antibiotics in the market are not successful against P. aeruginosa infections due to the pathogen’s ability to cause antibiotic resistance. Therefore, our prime objective is to formulate a liposomal-based anti-Pseudomonas vaccine that can produce long-lasting IgG antibodies and T cell memory. The vaccine we introduced accommodates several B cell epitopes of the outer-membrane protein F (OprF) of P. aeruginosa. The recombinant OprF antigen was conjugated with a Toll-like receptor 2/1 (TH1/TH2) agonist, Pam3CysSK4, and formulated into dipalmitoylphosphatidylcholine (DPPC)/cholesterol (Chol) liposomes. Our results show that the resulting vaccine conjugate can successfully produce high anti-OprF antibody titers in mice. These antibodies can also recognize intact P. aeruginosa bacteria and kill them with the support of rabbit complement and murine macrophage RAW264.7 cells. To further enhance the vaccine efficacy, the liposome was optimized with 2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycerol-3-phospho-(1′-rac-glycerol) (DMPG), Chol, Pam3CysSK4 and Quillaja Saponaria-derived saponin adjuvant QS21. The mice immunized with the modified vaccine elicit higher antibody titers and increased IgG2a antibodies. Overall, our liposomal vaccines were found to be highly effective at generating a balanced and robust TH1/TH2 response.

Supported by grants from NIH (R01AI148570)
Central nervous system IL-17R expression influences weight gain by altering food-intake and fat-mass accumulation
Authors: Fathi Payam , Ayala Julio E
Published: August 2023

Cytokines influence innate behaviors in mice and IL-17a may function within the hypothalamus to reduce food-intake. We examine contributions of nervous system IL-17R expression on mouse feeding behavior following obesogenic diet exposure.

We crossed IL-17Raf/f and Synapsin1-Cre or Nav1.8-Cre mice to generate pan-neuronal (Syn-CreIL-17Ra f/f) or peripheral neuronal- (Na-v1.8-CreIL17Ra f/f) IL-17R knockout mice, respectively. Body weights of male and female mice were measured over 10-weeks of 60% high-fat diet (HFD) feeding. 60% HFD fed male Syn-CreIL-17Ra f/f and IL-17Raf/f mice underwent metabolic phenotyping and body composition analyses. Syn-CreIL-17Ra f/f and IL-17Raf/f mice received CNS penetrant Nrf2 activator Dimethyl-Fumarate (DMF) to assess whether pharmacological inhibition of IL-17a secretion alters HFD induced weight gain.

Male and female Syn-CreIL-17Ra f/f but not Na-v1.8-CreIL17Ra f/f mice displayed a 15–20% reduction in bodyweight after 10-weeks of HFD feeding (male p=0.00046, female p=0.0061). DMF treatment did not impact body weight. Syn-CreIL-17Ra f/f trended towards a reduction in food-intake and meal numbers when compared to IL-17Raf/f mice. Syn-CreIL-17Ra f/f had a significant reduction in fat mass (total p=0.01, %total p=0.04) and lean mass (total p=0.03 and %total p=0.04).

In sum, loss of central but not peripheral nervous system expression of IL-17R robustly and significantly blunts weight gain following chronic exposure to an obesogenic diet. This decrease in body weight was associated with a decrease in total fat mass. Our data suggest that a population of IL-17R expressing neurons within the brain influence food-intake by modulating feeding behaviors associated with meal frequency.

NIH funding sources include F31-DK127728 and T32-DK07563
Cyanobacterial toxins and components induce expression of pro-inflammatory cytokines/chemokines and secretion of soluble cell receptors in a 3D skin cell model
Authors: Augustine Swinburne A J , Eason Tarsha N , Robillard Michelle C
Published: August 2023

Exposure to cyanobacterial toxins has been linked to several diseases and disorders including skin rashes and damage to the liver and kidneys. Previous research has shown that Microcystin-LR, the most common and toxic microcystin in fresh water, induces the NLRP3 inflammasome in response to administration to mice by oral gavage. In this study, we used a Reconstructed Human Epidermis to investigate immune system responses to cyanobacterial exposure. This 3D tissue model consisting of normal, human-derived epidermal keratinocytes was exposed to Spirulina, anatoxin-contaminated lake water, and phycocyanin. The cell supernatant was probed with a multiplex of 38 cytokines/chemokines and 14 soluble receptors to elucidate the immunological response. Our results revealed that: while most proteins were down-regulated, a number were up-regulated; the expression of many proteins were highly correlated and clustered together; and across the case study datasets, multiple cytokines/chemokines (G-CSF, GRO, IL-6, IL-8, and MCP-1) and soluble cell receptors (sCD30, sEGFR, sTNFRII and sVEGFR1, sVEGFR2 and sVEGFR3) were highly expressed when exposed to the toxins. Our findings correspond with preliminarily histological results showing tissue damage when the skin cells were exposed to the cyanobacterial components. This work shows great promise in the ability to examine the immune response to toxic exposures and aids in the development of techniques useful for studying the impact of harmful algal blooms on public health. Future studies will focus on mapping the immune pathway in response to harmful algal bloom exposure and studying the roles of each of the expressed cytokines and soluble cell receptors in the inflammatory response.

This work is supported by the United States Environmental Protection Agency.
A combination of TLR9 and STING agonists induces potent neopeptide-specific T cell immunity and improves ICB efficacy in tumor models
Authors: Hioki Kou , Castro Eiro Melisa D , Li Ling , Wilmsen Merel E P , Kiernan Caoimhe H , Brouwers-Haspels Inge , van Meurs Marjan , de Vos Kevin , Wildemans Marlous , Zhao Manzhi , de Wit Harm , Grashof Dwin G B , van de Werken Harmen J G , Mueller Yvonne M , Schliehe Christopher , Ishii Ken J , Katsikis Peter D
Published: August 2023

Immunotherapies such as immune checkpoint blockade (ICB) therapies have been developed in the last years as a promising strategy for the treatment of cancer, however there remains a need to improve their efficacy. The most critical factor that affects the efficacy of ICB therapies, is the frequency of tumor mutations, the associated neoantigens generated and the T-cell response against them. Therefore, it is expected that neoantigen vaccinations would improve ICB therapy efficacy by boosting the neoantigen-specific T cell response. The aim of this study is to develop an effective vaccine using a combination of TLR9 and STING agonists together with synthetic long peptides to induce a potent neoantigen-specific cytotoxic CD8+ T cell response. We first analyzed the efficacy of the vaccine formulation containing 20-mer OVA peptide and found that mice immunized with OVA peptide together with the adjuvant combination induced potent antigen-specific T cell responses and are capable of controlling tumor growth and improving survival in B16-F10-OVA tumor bearing mice. Moreover, we found that the vaccine together with OVA peptide is able to synergize with anti-PD-1 treatment in controlling tumor growth. Finally, we examined the immunogenicity of the vaccine formulation with neopeptides identified from two different tumor models. The vaccine formulation induced potent neoantigen-specific T cell responses in vivo. Our findings demonstrate that the vaccine formulation using the combination of TLR9 and STING agonists induces potent T cell immunity against synthetic long peptides and can serve as a promising immunogenic neoantigen vaccine platform.

Supported by the Department of Immunology of the Erasmus MC the Dutch Cancer Society (KWF grant 12837) and in part by a grant from International Joint Usage/Research Center, the Institute of Medical Science, the University of Tokyo, and Japan Agency for Medical Research and Development (AMED).
Effects of targeted epigenetic modifications on T – cell reprogramming
Authors: Norman Allison R , Ward Grace Anne , Zebley Caitlin C , Youngblood Ben A
Published: August 2023

Over the last decade there have been huge advances in DNA sequencing technology, allowing cancer patients to undergo genomic diagnosis within days of admission. Building upon genomic sequencing, epigenomic profiling approaches of tumor and infiltrating lymphocytes inform on the stage of the disease and immune response, but interpretation of these data remains hindered by issues of heterogeneity. Epigenetic programs, such as DNA methylation, play a vital role in the regulation of CD8+ T – cell development and differentiation. Here, we will investigate the effects of targeted epigenetic mechanisms on the function of effector memory T – cells. Hypomethylating agents, such as Decitabine, have been extensively used in myeloid malignancies including Acute Myeloid Leukemia and Myelodysplastic Syndromes; however, the impacts on T – cells have not been well studied. We hypothesize that the utilization of these epigenetic modifications will alter the function and phenotypic characteristics of the effector memory T – cells. These results will lead to new approaches for reprograming effector memory T – cells via removal of repressive DNA methylation. The outcome of this study will provide novel insight into our understanding of the epigenetic regulation of T – cells function; especially, how DNA methylation programs instilled during differentiation could be modified to improve T – cells function.
Molecular mechanisms of PBX1 regulation of CD4+ T cell inflammatory phenotypes
Authors: Park Yuk Pheel , Roach Tracoyia , So Sujung , Choi Seung-Chul , Ostrov David A , Yang Young , Morel Laurence
Published: August 2023

The pre-B cell leukemia homeobox1 (Pbx1) controls chromatin accessibility to a large number of genes in various cell types. Its dominant negative splice isoform, Pbx1-d that lacks the DNA and Hox-binding domains, is expressed more frequently in the CD4+ T cells from lupus-prone mice and SLE patients than healthy controls. We have shown that overexpression of Pbx1-d in CD4+ T cells impaired Treg cell homeostasis and expanded inflammatory CD4+ T cells. Here, we investigated the molecular mechanisms by which PBX1 regulates T cells. We showed that PBX1 is preferentially expressed in Treg cells over other CD4+ T cell subsets, and that it is negatively regulated by TCR activation. Furthermore, PBX1-D protein was less stable than the normal isoform, PBX1-B, in a pulse chase experiment. PBX1-B but not PBX1-D has two predictive ubiquitin binding sites, K292 and K293, located in the DNA binding domain. Mutation of these two sites reduced PBX1-B stability to a level similar to PBX1-D, suggesting that a loss of function in the lupus associated allele through the ubiquitin-proteasome dependent pathway. In addition, contrary to PBX1-B, PBX1-D is retained in the cytoplasm without the help of the MEIS or PREP1 co-factors, indicating different requirements for nuclear translocation. Overall, these findings suggest that multiple transcriptional and post-transcriptional mechanisms are responsible for PBX1-D loss of function and induction of CD4+ T cell inflammatory phenotypes in SLE.

This work was supported by National Institutes of Health Grants AI045050
ARYL HYDROCARBON RECEPTOR REGULATES GENES IN COLONIC EPITHELIAL CELLS TO MITIGATE COLITIS SEVERITY IN THE DSS-INDUCED MURINE MODEL
Authors: Saxena Archana , Mitchell Chandani , Staley Shanieka , Bogdon Raymond , Roark Kasie , Hailey Michele , Nagarkatti Mitzi , Nagarkatti Prakash , Busbee Brandon
Published: August 2023

Current treatment of ulcerative colitis (UC) leaves a clear unmet need to target the colonic epithelial cells (CECs). In our previous study we found that aryl hydrocarbon receptor (AhR) expression on CECs has a vital role during indole-3-carbinol (I3C)-mediated protection against colitis. However, the precise mechanism behind this remains unclear. We investigated how intestinal regulatory components were altered in the absence or presence of AhR in CECs during I3C treatment under colitis (mouse model) or colitis-like conditions (colonic organoid). We generated conditional AhR-CEC knockout mice (AV) and induced colitis using the dextran sodium sulfate (DSS) model. Our results showed that the AV mice lost the protective effects of I3C against colitis. AV mice had higher disease scores and increased inflammation compared to controls. I3C treated AV mice that had DSS induced colitis were not able to regulate the colitis-associated parameters, even though flow cytometry analysis revealed AV mice were still capable of increasing protective interleukin-22 (IL-22) production in the colon. These results prompted us to study the CEC-specific mechanism involved in I3C-mediated protection against colitis. Whole transcriptome analysis of RNA isolated from enriched CECs of experimental mice was performed and results showed significant altered expression of several microRNAs, mucins (muc2 and muc13), and tight junction proteins (Cld3 and Cld4) in AV mice when compared to controls. RT-PCR data and direct effects of I3C on CECs (immunostaining) using organoids validated these results. This data concluded that AhR-dependent signaling orchestrates I3C-mediated protection of colitis.

NIH 5P20GM103641
IFN-γ is Necessary but not Sufficient to mediate Activated CD4+ T cell-induced epithelial cell damage and death
Authors: Cruz Michelle A , Llyons Renae , Malhotra Riya , Meng Shanshan , Fuiji Satoru , Stappenbeck Thaddeus , Levine Alan D
Published: August 2023

Bidirectional communication between the intestinal epithelial barrier and the immune cells in the underlying lamina propria is critical to human health. This study investigates the impact of human T cell subtypes on intestinal epithelial cell phenotype and survival. Blood-derived CD4+ or CD8+ T cells were isolated from healthy human participants using negative magnetic separation and co-cultured with healthy human colonic epithelial stem cell spheroids suspended in Matrigel. A subset of T cells was activated by anti-CD3/CD28 cross-linking for 24 h prior to co-culture. Only activated CD4+ T cells (ACD4) caused significant epithelial cell damage and death as measured by histological scoring of brightfield microscopic images and zombie violet+ epithelial cells detected by flow cytometry.

The mechanism of cell damage and death was determined to be via soluble mediators, in that conditioned media from ACD4 but not activated CD8+ T cells (ACD8) significantly induced spheroid cell damage and death. A Luminex cytokine multiplex assay revealed that high levels of IFN-γ, IL-17, and TNF-α were produced by ACD4 as compared with ACD8. Neutralizing IFN-γ antibody completely protected spheroids from death induced by ACD4. In contrast, the addition of only IFN-γ protein did not induce spheroid cell death. This finding conflicts with currently published literature that indicates IFN-γ protein is able to induce spheroid cell death for both humans and mice. In addition, ACD8 also secrete IFN-γ, although to a lesser extent than ACD4, yet do not initiate spheroid cell death. These results indicate that there is a unique mixture of immune mediator released by ACD4s that mediate spheroid damage and death.

DP1 DA-037997

T32 GM007250
The roles of epidermal Blimp-1 in atopic dermatitis
Authors: Yang Hao-Wen , Wu Yen-Fei , Yang Shii-Yi , Lin Kuo-I
Published: August 2023

Keratinocytes are the main cell type constructing the epidermis of animals, which can further differentiate into corneocytes and express multiple innate immune receptors. B lymphocyte-induced maturation protein-1 (Blimp-1) plays a crucial rule in directing cornification of keratinocytes. We previously reported that keratinocyte-specific deletion of Prdm1, the gene encoding Blimp-1, induced spontaneous inflammation in the skin of adult mice. We further found that deletion of Prdm1 in keratinocytes in Balb/c genetic background caused hyper-production of type 2 cytokines and thymic stromal lymphopoietin (TSLP) in the skin, and staphylococci colonization, which are characteristics of atopic dermatitis (AD). Since K5-CreER+Prdm1f/f (CKO) mice in Balb/c background exhibited AD-like symptoms, we wondered whether they are more susceptible to the disease progression in a murine AD model induced by MC903, a vitamin D3 analog enhancing TSLP production by keratinocytes. We found that CKO mice showed more immune cell infiltration in ear skin, developed more severe epidermal hyperplasia, and induced more TSLP secretion in the skin following topical application of MC903. Recent studies showed that staphylococci colonization activated the transcription of endogenous retroviruses (ERVs) in murine skin, while excess ERV expression led to skin inflammation. As staphylococci colonized on our CKO mouse skin, we hypothesized that enhanced ERV expression may also link to skin inflammation of CKO mice. Indeed, both in vivo and in vitro deletion of Prdm1 in keratinocytes resulted in elevated levels of ERV transcripts. We are studying the contribution of elevated ERV transcripts in Blimp-1-reduction-mediated AD-like skin inflammation.
IgE production and stability regulated by Glycosylation in vivo
Authors: Roy Avishek , Meregini Steve , Chen Zhenglan , Nair-Gill Evan , Ludwig Sara , Russell Jamie , Quan Jiexia , Hildebrand Sara , Moresco Eva , Yue Tao , Beutler Bruce , SoRelle Jeffrey
Published: August 2023

Rationale

Essential functions of IgE have been attributed to specific glycosylations such as oligomannose (Fcer1a binding) and sialylation (pathogenicity). However, no study has examined how glycosylation affects IgE stability in vivo.

Methods

Forward genetic screening for IgE specific phenotypes was performed using N-ethyl-N-nitrosourea (ENU) mutagenized mice. One low IgE phenotype, named benadryl, linked to a mutation in Mpi, which is essential for N-linked glycosylation. Benadryl was validated by CRISPR knock-in . Immunological studies were performed to determine the causative mechanism (passive/ active anaphylaxis, mast cell analysis, conditional knockout in T, B and dendritic cells, antibody production in vivo and in vitro, using IgE-Venus, IgE-KO, and IL-4 GFP mice).

Results

Benadryl mice had low total and papain-specific IgE responses with resistance to active anaphylaxis. In vitro and in vivo mast cell activation studies resulted in no difference in response to passive sensitization anaphylaxis. Both, in vivo and in vitro cellular staining in the conditional knockout showed no decrease in IgE B cells. IgE from Mpi deficient mice was produced and secreted from B cell culture. When de-glycosylated IgE was injected to IgE KO mice, it was cleared 10-fold faster than native IgE.

Conclusions

Glycosylation status determines not only IgE functionality but in vivo stability. Mpi deficient mice produce IgE hypoglycosylated in nature and therefore it is cleared faster escaping anaphylaxis. Modulating IgE glycosylation represents a novel way to reduce IgE below pathogenic levels.

NA
AMBRA1 control immune-specific gene expression in T lymphocytes
Authors: Yao Yikun , Lenardo Michael
Published: August 2023

Autoimmune lymphoproliferative syndrome (ALPS) is a disease characterized by the dysregulation of cell death in lymphocytes, resulting in symptoms such as hepatomegaly, splenomegaly, and anemia. While most patients with ALPS have a mutation in the FAS gene or within the FAS pathway, which helps mediate cell death, some patients who present with ALPS-like symptoms do not have known causative gene mutations. To identify other genes that confer resistance to cell death, and thus may play a role in diseases such as ALPS, we performed a whole genome CRISPR screen of Fas-mediated cell death and identified AMBRA1, a protein previously studied in autophagy and E3 ubiquitin ligase activity. We verified the screen results and indeed found that AMBRA1 KO cells resist Fas-mediated cell death. To identify the mechanism of how AMBRA1 contributes to cell death resistance, we performed cycloheximide chase assays, qPCR, and proteomic analysis and confirmed that AMBRA1 regulates FAS expression. Furthermore, we found that AMBRA1 itself can be upregulated by TCR stimulation through the CD28-PI3K-mTORC1 pathway. Mass spectrometry analysis and subsequent experiments also confirmed that AMBRA1 regulates other immune genes. Together, these data establish a novel function for AMBRA1 as a critical regulator in T cells and unveil new insights into how AMBRA1 may contribute to ALPS and dysregulated cell death in other immune diseases. This research was supported by the Intramural Research Program of the NIH.
NK-92-CD16A cell externalization of CD107a for detection of antigen-specific antibodies able to support ADCC: sensitive discrimination of antibody a-fucosylation
Authors: Amaya Judith B Cruz , Walcheck Bruce , Smith-Gagen Julie , Lombardi Vincent , Hudig Dorothy
Published: August 2023

Antibody-dependent cell-mediated cytotoxicity (ADCC) by natural killer (NK) lymphocytes kills cells infected with enveloped viruses. Antiviral ADCC has three components: antigen-specific IgG1 or IgG3; effector cells with CD16A that recognize the Fc of IgG; and antigenic proteins in the plasma membranes of infected cells. Afucosylated antibodies support ADCC better than fucosylated antibodies; afucosylated fractions vary among individuals. Current assays that detect afucosylated antibodies to specific viruses involve antigen-specific antibody purification and mass spectrometry. We are developing clinical bioassays to detect antibodies that will support ADCC and assess their extents of afucosylation. Our pilot assay uses externalization of CD107a by NK-92-CD16A cells, this’ occurs when NK cells recognize cell-bound antibodies. We used two humanized monoclonal antibodies on CD20, GA101 WT or glycoengineered GE, 10% or ~50% afucosylated, with Raji cells. Cellular CD107a externalization was more sensitive than AF-647 donkey anti-IgG to detect total cell-bound antibody. The effective concentrations (EC50s) of the two antibodies for CD107a externalization differed by >15-fold. The more afucosylated GA101-GE was more detectable, at 0.2 ng/ml (=/< 700 Abs/Raji cell). CD107a externalization increased with excess antibody-bearing cells. Short 37 °C incubation (40 minutes) optimized detection of ADCC over intrinsic NK activity. Controls of GA101-GE alone and 56 °C heated human serum lacked effects on CD107a externalization. The assay appears compatible with formaldehyde treatment before NK cell staining. This sensitive bioassay appears suitable for assess serum samples for antiviral antibodies that will support ADCC.

UNR Foundation C.E. Hudig Fund.