Active Complex Research Articles

Time-dependent complexity characterisation of activity patterns in patients with Chronic Fatigue Syndrome

BackgroundChronic Fatigue Syndrome patients suffer from symptoms that cannot be explained by a single underlying biological cause. It is sometimes claimed that these symptoms are a manifestation of a disrupted autonomic nervous system. Prior works studying this claim from the complex adaptive systems perspective, have observed a lower average complexity of physical activity patterns in chronic fatigue syndrome patients compared to healthy controls. To further study the robustness of such methods, we investigate the within-patient changes in complexity of activity over time. Furthermore, we explore how these changes might be related to changes in patient functioning.MethodsWe propose an extension of the allometric aggregation method, which characterises the complexity of a physiological signal by quantifying the evolution of its fractal dimension. We use it to investigate the temporal variations in within-patient complexity. To this end, physical activity patterns of 7 patients diagnosed with chronic fatigue syndrome were recorded over a period of 3 weeks. These recordings are accompanied by physicians’ judgements in terms of the patients’ weekly functioning.ResultsWe report significant within-patient variations in complexity over time. The obtained metrics are shown to depend on the range of timescales for which these are evaluated. We were unable to establish a consistent link between complexity and functioning on a week-by-week basis for the majority of the patients.ConclusionsThe considerable within-patient variations of the fractal dimension across scales and time force us to question the utility of previous studies that characterise long-term activity signals using a single static complexity metric. The complexity of a Chronic Fatigue Syndrome patient’s physical activity signal does not suffice to characterise their high-level functioning over time and has limited potential as an objective monitoring metric by itself.

Characterization of the transcriptionally active form of dephosphorylated DctD complexed with dephospho-IIAGlc.

Bacterial enhancer-binding proteins (bEBPs) acquire a transcriptionally active state via phosphorylation. However, transcriptional activation by the dephosphorylated form of bEBP has been observed in DctD, which belongs to Group I bEBP. The formation of a complex between dephosphorylated DctD (d-DctD) and dephosphorylated IIAGlc (d-IIAGlc) is a prerequisite for the transcriptional activity of d-DctD. In the present study, characteristics of the transcriptionally active complex composed of d-IIAGlc and phosphorylation-deficient DctD (DctDD57Q) of Vibrio vulnificus were investigated in its multimeric conformation and DNA-binding ability. DctDD57Q formed a homodimer that could not bind to the DNA. In contrast, when DctDD57Q formed a complex with d-IIAGlc in a 1:1 molar ratio, it produced two conformations: dimer and dodecamer of the complex. Only the dodecameric complex exhibited ATP-hydrolyzing activity and DNA-binding affinity. For successful DNA-binding and transcriptional activation by the dodecameric d-IIAGlc/DctDD57Q complex, extended upstream activator sequences were required, which encompass the nucleotide sequences homologous to the known DctD-binding site and additional nucleotides downstream. This is the first report to demonstrate the molecular characteristics of a dephosphorylated bEBP complexed with another protein to form a transcriptionally active dodecameric complex, which has an affinity for a specific DNA-binding sequence.IMPORTANCEResponse regulators belonging to the bacterial two-component regulatory system activate the transcription initiation of their regulons when they are phosphorylated by cognate sensor kinases and oligomerized to the appropriate multimeric states. Recently, it has been shown that a dephosphorylated response regulator, DctD, could activate transcription in a phosphorylation-independent manner in Vibrio vulnificus. The dephosphorylated DctD activated transcription as efficiently as phosphorylated DctD when it formed a complex with dephosphorylated form of IIAGlc, a component of the glucose-phosphotransferase system. Functional mimicry of this complex with the typical form of transcriptionally active phosphorylated DctD led us to study the molecular characteristics of this heterodimeric complex. Through systematic analyses, it was surprisingly determined that a multimer constituted with 12 complexes gained the ability to hydrolyze ATP and recognize specific upstream activator sequences containing a typical inverted-repeat sequence flanked by distinct nucleotides.

‘You certainly don’t get promoted for just teaching’: experiences of education-focused academics in research-intensive universities

ABSTRACT Changes in drivers of academic roles within higher education institutions globally have resulted in increased proportions of academics in education focused (EF) posts. International and UK research suggests that EF academics can experience dissatisfaction with career progression and the perceived value of their work, including those in research-intensive universities. Previous UK research was conducted prior to the introduction of the TEF which has altered the landscape. Therefore, it was timely to examine the current experience of EF academics in research-intensive universities through a theoretical lens to understand barriers and facilitators to career progression. This interview-based study used two theoretical frameworks, Feldman and Ng’s Framework for Career Mobility, Embeddedness, and Success and Kanter’s theory of Power within organisations to explore the experiences of 43 EF academics across 12 research-intensive UK universities. Four contract types were identified, some of which allowed promotion. Three broad themes were derived from the data, including (1) Lack of agreement on the definition of education-focused academic roles, (2) Level of value and appreciation of educational expertise and the impact on education-focused academics, (3) Career development opportunities for education-focused academics. Recommendations to further enhance the experience and career progression for EF academics in research-intensive universities further include; ensuring transparency in recruitment into EF posts as to whether career development is possible within that post, the need to continue the sector-wide discussion on the definition of EF roles that recognises the complexity and diversity of activity and continued work to value and recognise appropriately educational expertise.

Diversity, spatial distribution and evolution of inactive and weakly active hydrothermal deposits in the TAG hydrothermal field

Introduction: Although, there is an increasing focus on inactive or extinct seafloor massive sulfide (SMS) deposits driven by the possibility of marine mining, only few studies have been devoted to them so far. The Trans-Atlantic Geotraverse (TAG) hydrothermal field is probably one of the best-studied hydrothermal systems even if the relict SMS deposits known since the mid-1980s have not been thoroughly explored.Objectives: The main objective of this study was to describe the characteristics of these so-called inactive sites.Methods: During four different expeditions, we acquired high-resolution acoustic data and performed numerous human occupied vehicle (HOV) dive operations including extensive rock sampling and in-situ temperature measurements.Results and Discussion: We discovered thirteen new hydrothermal mounds including six large (i.e. > 5,000 m2) deposits making the TAG hydrothermal field one of the largest accumulation of hydrothermal materials (21.1 Mt) known on the seafloor. However, copper and zinc grades of the largest SMS deposits remain low (i.e. < 1.4 wt%) even compared to on-land volcanogenic massive sulfide deposits. Additionally, eight areas of diffuse hydrothermal fluid flow were identified challenging the presumed inactivity of these SMS deposits and, for the first time, emphasizing the importance of low temperature (LT) hydrothermal activity in whole the TAG field. Inactive and weakly active SMS deposits exhibit a large diversity of surface mineralization (e.g. sulfides, Fe-Mn mineralization, jasper) illustrating complexity of hydrothermal activities but also different ageing history. Several mounds no longer have visible sulfide chimneys and are covered by a widespread layer of manganese and iron oxyhydroxides attesting the longevity of diffuse fluid flow at specific locations even long after last high-temperature (HT) hydrothermal activity has ceased. This contrasts with SMS deposits that are devoid of extensive LT precipitates but characterized by standing or topped sulfide chimney indicating a relatively abrupt cessation of HT hydrothermal activity.Conclusion: Together these results allow us to propose evolution models to explain the diversity of active, weakly active and inactive SMS deposits in the TAG hydrothermal field.

ZMIZ1 enhances ERα-dependent expression of E2F2 in breast cancer.

The estrogen receptor-α (ER) drives 75% of breast cancers. On activation, the ER recruits and assembles a 1-2 MDa transcriptionally active complex. These complexes can modulate tumour growth, and understanding the roles of individual proteins within these complexes can help identify new therapeutic targets. Here, we present the discovery of ER and ZMIZ1 within the same multi-protein assembly by quantitative proteomics, and validated by proximity ligation assay. We characterise ZMIZ1 function by demonstrating a significant decrease in the proliferation of ER-positive cancer cell lines. To establish a role for the ER-ZMIZ1 interaction, we measured the transcriptional changes in the estrogen response post-ZMIZ1 knockdown using an RNA-seq time-course over 24 h. Gene set enrichment analysis of the ZMIZ1-knockdown data identified a specific delay in the response of estradiol-induced cell cycle genes. Integration of ENCODE data with our RNA-seq results identified that ER and ZMIZ1 both bind the promoter of E2F2. We therefore propose that ER and ZMIZ1 interact to enable the efficient estrogenic response at subset of cell cycle genes via a novel ZMIZ1-ER-E2F2 signalling axis. Finally, we show that high ZMIZ1 expression is predictive of worse patient outcome, ER and ZMIZ1 are co-expressed in breast cancer patients in TCGA and METABRIC, and the proteins are co-localised within the nuclei of tumour cell in patient biopsies. In conclusion, we establish that ZMIZ1 is a regulator of the estrogenic cell cycle response and provide evidence of the biological importance of the ER-ZMIZ1 interaction in ER-positive patient tumours, supporting potential clinical relevance.

Integration of electronic project management software in an investigational drug service pharmacy within an academic medical center.

An investigational drug services (IDS) pharmacy plays a vital role in supporting clinical trial research by ensuring the safe and efficient management of investigational products. This article describes the implementation of an electronic project management software to improve an IDS pharmacy's study protocol work. The article describes the implementation of the software and how this approach addressed specific challenges, including project oversight, process standardization, documentation, reporting, accountability, and intrateam communication. We describe an electronic project management software system used to streamline and standardize the work associated with study protocols. This software provides an organized and customizable workspace to manage tasks associated with each study protocol. The software automates task creation, tracks progress, and ensures comprehensive record keeping. Additionally, the software fosters effective communication within the team and offers real-time reporting to assess team productivity and progress. We have observed improved consistency, enhanced revenue, including approximately $18,000 in additional fee capture, and increased collaboration among pharmacy team members. Implementing an electronic project management software has proven highly beneficial in the IDS pharmacy. The software has significantly improved workflow efficiency by addressing challenges in study protocol management. While initial setup and training required time and resources, the long-term benefits in project oversight, collaboration, and revenue capture justify the investment. An electronic project management software is a valuable tool in managing the complexity of study protocol activities and supports the pharmacy's crucial role in advancing clinical research.

Age-dependent loss of Crls1 causes myopathy and skeletal muscle regeneration failure

Skeletal muscle aging results in the gradual suppression of myogenesis, leading to muscle mass loss. However, the specific role of cardiolipin in myogenesis has not been determined. This study investigated the crucial role of mitochondrial cardiolipin and cardiolipin synthase 1 (Crls1) in age-related muscle deterioration and myogenesis. Our findings demonstrated that cardiolipin and Crls1 are downregulated in aged skeletal muscle. Moreover, the knockdown of Crls1 in myoblasts reduced mitochondrial mass, activity, and OXPHOS complex IV expression and disrupted the structure of the mitochondrial cristae. AAV9-shCrls1-mediated downregulation of Crls1 impaired muscle regeneration in a mouse model of cardiotoxin (CTX)-induced muscle damage, whereas AAV9-mCrls1-mediated Crls1 overexpression improved regeneration. Overall, our results highlight that the age-dependent decrease in CRLS1 expression contributes to muscle loss by diminishing mitochondrial quality in skeletal muscle myoblasts. Hence, modulating CRLS1 expression is a promising therapeutic strategy for mitigating muscle deterioration associated with aging, suggesting potential avenues for developing interventions to improve overall muscle health and quality of life in elderly individuals.

АНКЕТИРОВАНИЕ СРЕДИ СТУДЕНТОВ ОШСКОГО ГОСУДАРСТВЕННОГО УНИВЕРСИТЕТА О УПОТРЕБЛЕНИИ ЭНЕРГЕТИЧЕСКИХ НАПИТКОВ

. In modern society there is a problem associated with constant evolution and acceleration of the pace of life. The increasing complexity of human activity is accompanied by increasing demands on the results of his work. This leads to a decrease in the attention paid by individuals to their lifestyle, which in turn manifests itself in a decrease in activity, partly due to the proliferation of computers, reduced leisure time, changes in sleep patterns and disruption of healthy eating principles. In response to such changes, people are turning to the consumption of energy drinks, aimed at increasing alertness and stimulating physical or mental activity, temporarily restoring energy and activating the nervous system. In this regard, the use of tonic drinks is popularized among the population of Kyrgyzstan. A particular increase in consumption is observed among young people. However, excessive consumption of such drinks can pose serious dangers. A survey was conducted among students of Osh State University to study the practice of consuming energy drinks. Students were studied in different categories: by age, by gender and by university faculties. The results showed that awareness of the dangers of these drinks varied among students of different faculties, with medical students being more likely to recognize the potential risks. By age category, young people drink much more often than other respondents; it was also noted that women rarely resort to drinking energy drinks than men. In addition, links have been found between the consumption of energy drinks and dental status: students who drink energy drinks are more likely to experience problems with caries and the need for tooth extraction, which may indicate a negative impact of these drinks on health.

INFORMATION TECHNOLOGY PLATFORM FOR ARMS TRADE

Based on the postulate that the arms trade is generally a complex, specific and multifaceted field of activity, which includes the production, distribution, sale and use of various types of weapons and military equipment. In this context, the authors of the article proposed the development of an innovative information technology platform for accounting and analysis of arms trade processes, which takes into account the peculiarities of domestic legislation. It was noted that the use of blockchain technology on the information and technological platform of weapons sales is aimed at increasing security and tracking the full set of transactions, reducing risks and ensuring maximum transparency of processes. It is assumed that each stage of the life cycle of weapons, starting from design, production and ending with sale, will be recorded by blockchain methods and means. Information about the manufacturer, the date of manufacture, transportation, purchase and acceptance of the weapon by the customer will be recorded in the blocks. With the use of an information technology platform, the validity of licenses will be checked with each transaction, ensuring the legality of the sale and taking into account the unique characteristics of each weapon unit. The information technology platform will store information about transactions, including price, date and participants, smart contracts, etc. Development of an information technology platform that is conveniently integrated into the system of international standards and regulators, as well as meets numerous specific requirements and restrictions on arms trade. The authors of the article claim that the use of cryptography in blockchain technology provides reliable encryption and protection of confidential information, which is critical in a sensitive subject area such as the arms trade, and also contributes to a significant increase in the level of security, transparency and trust among participants in this specific industry. The conducted analysis confirms the expediency of using the potential of blockchain technology to optimize and increase the efficiency of the accounting of arms trade processes, as well as to ensure the reliability and safety of the functioning of the innovative information technology platform.

Examination of Primary and Secondary Metabolites Associated with a Plant-Based Diet and Their Impact on Human Health.

The consumption of plant-based diets has become a burgeoning trend, and they are increasingly consumed globally owing to their substantial energy intensity and dietetic advantages. Plants possess numerous bioactive components that have been recognized to exhibit manifold health-promoting assets. Comprehension of the synthesis of these primary and secondary metabolites by plants and their method of action against several chronic illnesses is a significant requirement for understanding their benefits to human health and disease prevention. Furthermore, the association of biologically active complexes with plants, humans, disease, medicine, and the underlying mechanisms is unexplored. Therefore, this review portrays various bioactive components derived from plant sources associated with health-promoting traits and their action mechanisms. This review paper predominantly assembles proposed plant-derived bioactive compounds, postulating valuable evidence aimed at perceiving forthcoming approaches, including the selection of potent bioactive components for formulating functional diets that are effective against several human disorders. This meticulous evidence could perhaps provide the basis for the advanced preemptive and therapeutic potential promoting human health. Hence, delivery opens possibilities for purchasers to approach the lucrative practice of plants as a remedy, produce novel products, and access new marketplaces.

Application of Machine Learning-Based K-means Clustering for Financial Fraud Detection

In today's increasingly digital financial landscape, the frequency and complexity of fraudulent activities are on the rise, posing significant risks and losses for both financial institutions and consumers. To effectively tackle this challenge, this paper proposes a machine learning-based K-means clustering method to enhance the accuracy and efficiency of financial fraud detection. By clustering vast amounts of financial transaction data, we can identify anomalous patterns and behaviors in a timely manner, thereby detecting potential fraud. Compared to traditional rule-based detection methods, machine learning-based approaches better adapt to ever-evolving fraud techniques and patterns while improving flexibility and precision in detection. Moreover, K-means clustering also aids in optimizing resource allocation within financial institutions by enabling focused monitoring and prevention efforts in high-risk areas, thus effectively mitigating the impact of fraud on the overall financial system. In summary, the machine learning-based K-means clustering method holds promising prospects for application in the field of financial fraud detection as it strives to establish a more secure and reliable transaction environment for the finance industry.

Identification of Repurposed FDA Drugs by Targeting Sclerostin via the Wnt Pathway for Alveolar Bone Formation

Objective Natural wingless-related integration site (Wnt) pathway antagonist sclerostin (SOST) has attracted much attention because unusual bone illnesses characterized by the increased bone mass result from its absence of action. The Wnt ligand is prevented from attaching to the Frizzled family receptor when SOST is present. In the active destruction complex, -catenin is phosphorylated. -Catenin molecules do not enter the nucleus and are broken down by a proteasome. As a result, Wnt-responsive genes are not activated, which lowers bone formation and raises bone resorption. A humanized monoclonal antibody called romosozumab binds to and inhibits SOST with significant cardiac side effects. As a result, the current study's objective is to find and screen Food and Drug Administration (FDA) medications that target SOST. Materials and Methods SOST's structure was retrieved from Protein Data Bank (PDB) (ID: 6l6r). Pharmacophore modeling and molecular operating environment-based virtual testing of FDA-approved medicines. Using the Desmond program, docking and molecular dynamics simulations were performed. Results Our findings revealed medications with FDA approval (ZINC000253387843) Amphotericin B. The stability and receptor–ligand interactions are pretty substantial, as demonstrated by the findings of docking and Molecular dynamics simulations, which have a docking score of −7.3 k/mol and root mean square deviation stability at 40 nanoseconds, respectively. Conclusion The suggested pharmacological therapy shows promise since it uses the Wnt pathway to target the primary bone formation mechanism. However, additional prospective studies are required to apply the available data to clinical practice.

Insight into the influence of natural deep eutectic solvents on the extraction of phenolic compounds from poplar type propolis: Composition and in vitro biological activity

Natural deep eutectic solvents (NADESs) have been considered promising to replace traditional volatile and toxic organic solvents for the extraction of biologically active substances from natural sources. This work applied an efficient and ethanol-exclusion strategy for extraction of phenolic compounds from poplar type propolis using five known NADESs (lactic acid:1,2-propanediol 1:1, lactic acid:fructose 5:1, choline chloride:1,2-propanediol 1:3, choline chloride:1,2-propanediol:water 1:1:1 and betaine:malic acid:water 1:1:6). The selected NADESs' extractability was evaluated by measuring the concentrations of total phenolics and total flavones and flavonols in the propolis extracts obtained, which qualitative chemical composition was further determined in detail by gas chromatography-mass spectrometry (GC-MS) analysis. It demonstrated that the chemical profiles of NADES and 70% ethanolic propolis extracts are similar. To expand the knowledge about the role of the applied solvents in the poplar propolis extraction process, the in vitro antimicrobial, cytotoxic and genotoxic activity of both NADESs and liquid NADES extracts were evaluated. The results revealed that the use of the selected NADESs as an extraction media for phenolic compounds from poplar propolis not only delivered a good extraction yield in some cases, but generally led to the preservation of propolis extracts' biological activity and even to the enhancement of their antimicrobial effect in comparison with the hydroethanolic one. Besides, the tested NADESs except for lactic acid:fructose and betaine:malic acid:water exerted low to negligible toxicity against normal cells treated and apart from lactic acid:fructose the remaining solvents demonstrated concentration-dependent moderate to subtle genotoxicity. There is a probability that not the supramolecular structure of the NADESs, but their components, played a key role for the observed biological effects. The present study has demonstrated an alternative approach for extracting the biologically active complex from poplar type propolis using NADESs, which could be useful for further pharmaceutical and cosmeceutical applications.

The adaptor protein ECAP, the corepressor LEUNIG, and the transcription factor BEH3 interact and regulate microsporocyte generation in Arabidopsis.

Histospecification and morphogenesis of anthers during development in Arabidopsis (Arabidopsis thaliana) are well understood. However, the regulatory mechanism of microsporocyte generation at the pre-meiotic stage remains unclear, especially how archesporial cells are specified and differentiate into 2 cell lineages with distinct developmental fates. SPOROCYTELESS (SPL) is a key reproductive gene that is activated during early anther development and remains active. In this study, we demonstrated that the EAR motif-containing adaptor protein (ECAP) interacts with the Gro/Tup1 family corepressor LEUNIG (LUG) and the BES1/BZR1 HOMOLOG3 (BEH3) transcription factor to form a transcription activator complex, epigenetically regulating SPL transcription. SPL participates in microsporocyte generation by modulating the specification of archesporial cells and the archesporial cell-derived differentiation of somatic and reproductive cell layers. This study illustrates the regulation of SPL expression by the ECAP-LUG-BEH3 complex, which is essential for the generation of microsporocytes. Moreover, our findings identified ECAP as a key transcription regulator that can combine with different partners to regulate gene expression in distinct ways, thereby facilitating diverse processes in various aspects of plant development.

Evaluation of wound healing potential of lawsone from Lawsonia innermis and its organometallic complexes in rats

Aim of the studyTo study the utility of Lawsone along with its zinc and copper complexes for wound healing activity using excision and incision wound models in rats. MethodsThe complex of plant nathoquinone from Lawsonia inermis called Lawsone viz Law and the metals viz. copper and zinc were synthesized. Molecular Docking (with GSK 3β protein) and Molecular Dynamic studies were carried out. The wound healing activity was evaluated using excision and incision wound models in Albino Wistar rats. The six groups comprised disease control, vehicle control, and standard, groups treated with Lawsone (Law) and its Zn Complex (ZnLaw) and copper complex (CuLaw). The progress of excision wound healing was monitored through wound contraction and biochemical and antioxidant parameters along with histopathology of granulation tissue, while for incision wounds, breaking and tensile strength were determined. ResultsThe molecular docking studies revealed that ZnLaw has the highest affinity with GSK 3β protein. The Molecular Dynamic study determined the stability of Law and its metal complexes with GSK 3β protein and was found to be stable in the environment. The test compounds Law, ZnLaw, and CuLaw displayed significant (p<0.05) wound contraction. There was a significant (P< 0.01) decrease in the period of epithelisation as compared to vehicle control. ZnLaw showed maximum reduction in the period of epithelisation (16.03±0.942). There was a significant improvement in the antioxidant parameters from the granulation tissues of wounds treated with Law, ZnLaw, and CuLaw. The metal complexes of Law viz. ZnLaw and CuLaw showed a remarkable increase in hydroxyproline and hexosamine content as compared to the control. Histopathological findings indicated re-epithelisation, neovascularization, and increased collagen deposition as compared to the disease control. The incision model also revealed a significant (P< 0.001) increase in the tensile strength and breaking strength of the healed tissue as compared to the control. ConclusionLaw and its Zinc and copper complexes exhibited good potential in healing of both incision and excision wounds.

Molecular mechanism of muscarinic acetylcholine receptor M3 interaction with Gq

Muscarinic acetylcholine receptor M3 (M3) and its downstream effector Gq/11 are critical drug development targets due to their involvement in physiopathological processes. Although the structure of the M3-miniGq complex was recently published, the lack of information on the intracellular loop 3 (ICL3) of M3 and extensive modification of Gαq impedes the elucidation of the molecular mechanism of M3-Gq coupling under more physiological condition. Here, we describe the molecular mechanism underlying the dynamic interactions between full-length wild-type M3 and Gq using hydrogen-deuterium exchange mass spectrometry and NanoLuc Binary Technology-based cell systems. We propose a detailed analysis of M3-Gq coupling through examination of previously well-defined binding interfaces and neglected regions. Our findings suggest potential binding interfaces between M3 and Gq in pre-assembled and functionally active complexes. Furthermore, M3 ICL3 negatively affected M3-Gq coupling, and the Gαq AHD underwent unique conformational changes during M3-Gq coupling.

Fluorescent reporters give new insights into antibiotics-induced nonsense and frameshift mistranslation

We developed a reporter system based on simultaneous expression of two fluorescent proteins: GFP as a reporter of the capacity of protein synthesis and mutated mScarlet-I as a reporter of translational errors. Because of the unique stop codons or frameshift mutations introduced into the mScarlet-I gene, red fluorescence was produced only after a mistranslation event. These reporters allowed us to estimate mistranslation at a single cell level using either flow cytometry or fluorescence microscopy. We found that laboratory strains of Escherichia coli are more prone to mistranslation compared to the clinical isolates. As relevant for uropathogenic E. coli, growth in human urine elevated translational frameshifting compared to standard laboratory media, whereas different standard media had a small effect on translational fidelity. Antibiotic-induced mistranslation was studied by using amikacin (aminoglycoside family) and azithromycin (macrolide family). Bactericidal amikacin induced preferably stop-codon readthrough at a moderate level. Bacteriostatic azithromycin on the other hand induced both frameshifting and stop-codon readthrough at much higher level. Single cell analysis revealed that fluorescent reporter-protein signal can be lost due to leakage from a fraction of bacteria in the presence of antibiotics, demonstrating the complexity of the antimicrobial activity.

Abstract 3011: The loss of an orphan nuclear receptor NR2E3 augments Wnt/β-Catenin signaling via epigenetic dysregulation that links to the Sp1-β catenin-p300 interactions in hepatocellular carcinoma

Abstract Hepatocellular carcinoma (HCC) stands as the fourth leading cause of global cancer-related mortality due to late-stage detection, tumor heterogeneity, and drug resistance. The pivotal role of epigenetic factors in HCC development and progression necessitates the identification of critical epigenetic regulators. The orphan nuclear receptor NR2E3 (Nuclear receptor subfamily 2 group E, Member 3), primarily associated with retinal development, has emerged as a potential tumor suppressor. Earlier studies showed that the loss of NR2E3 leads to epigenetic repression of the estrogen receptor α (ER) through the involvement of Lysine Specific histone Demethylase 1 (LSD1). NR2E3 also forms a transcriptional complex with Sp1, governing the aryl hydrocarbon receptor (AHR) expression, a crucial factor in liver xenobiotic metabolism. In vivo, NR2E3 ablation results in p53 inactivation and severe liver injuries when exposed to liver toxicants, highlighting its role in activating p53. Nonetheless, the precise tumor suppressive and epigenetic role of NR2E3 in HCC remains unclear. HCC patients expressing low NR2E3 exhibit unfavorable clinical outcomes, aligning with heightened activation of the Wnt/β-catenin signaling pathway. The murine HCC models utilizing NR2E3 knockout mice show accelerated liver tumor formation and progression accompanied by enhanced activation of the Wnt/β-catenin signaling pathway and inactivation of the p53 signaling pathway. At a cellular level, losing NR2E3 increases the acquisition of aggressive cancer cell phenotype and tumorigenicity and upregulates vital genes in the Wnt/β-catenin pathway with enhanced chromatin accessibility. This event is mediated through the increased formation of active transcription complex involving Sp1, β-catenin, and p300, a histone acetyltransferase, on the promoters of target genes. Correspondingly, treatment of p300 inhibitor C646 decreased the expression of critical genes in the Wnt/β-catenin signaling pathway. These findings demonstrate that the loss of NR2E3 promotes Wnt/β-catenin signaling activation at cellular, organismal, and clinical levels. In summary, NR2E3 is a novel tumor suppressor that maintains epigenetic homeostasis, thereby preventing activation of Wnt/β-catenin signaling that promotes HCC formation. Citation Format: Yuet-Kin Leung, Sung-Gwon Lee, Jiang Wang, Shuk-Mei Ho, Nancy Rusch, Chungoo Park, Kyounghyun Kim. The loss of an orphan nuclear receptor NR2E3 augments Wnt/β-Catenin signaling via epigenetic dysregulation that links to the Sp1-β catenin-p300 interactions in hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3011.

Abstract 3239: Targeting the GAS41 YEATS domain in non-small cell lung cancer with novel small molecules

Abstract Background: Glioma amplified sequence 41 (GAS41) belongs to the Yaf9, ENL, AF9, Taf14, and Sas5 (YEATS) domain family of epigenetic reader proteins. This domain preferentially recognizes acetylated lysine residues on chromatin to facilitate gene expression by recruiting chromatin remodeling and transcription activating complexes. The YEATS4 gene encoding for GAS41 protein is frequently amplified in NSCLC patient samples relative to normal lung tissue, and GAS41 protein is overexpressed in NSCLC cells. In knockdown experiments using NSCLC xenograft mouse models, GAS41 has been shown to be essential for NSCLC tumor growth. In mutagenesis studies, the YEATS domain residues responsible for acetylation reader activity were found to be essential for NSCLC cellular proliferation and tumor progression. Thus, the YEATS domain of GAS41 represents an attractive target for the development of small-molecule inhibitors as potential therapeutics for NSCLC. Results: We developed the first-in-class small-molecule inhibitors for the GAS41 YEATS domain. Using an NMR-based fragment screen, we identified a hit with low millimolar binding affinity. Structure-based drug design followed by extensive medicinal chemistry optimization yielded the potent and selective inhibitor DLG-369 with sub-micromolar inhibition of the GAS41 YEATS domain interaction with acylated histones. Using the NanoBRET platform, we showed that DLG-369 selectively disrupts the association of GAS41 YEATS with chromatin in mammalian cells. Furthermore, DLG-369 inhibits proliferation of NSCLC cell lines without affecting normal lung fibroblasts. In transcriptomic experiments, DLG-369 treatment regulates expression of multiple genes in NSCLC cells, including upregulation of the critical tumor suppressor genes CDKN1A and CYR61 and downregulation of chemokine genes CXCL5 and CXCR4 associated with NSCLC progression. Conclusions: We developed DLG-369, a potent inhibitor of the GAS41 YEATS domain with activity in NSCLC cellular models. DLG-369 potently binds to the YEATS domain acetylation reader pocket and disrupts GAS41 recognition of acetylation in cells. DLG-369 potently reduces proliferation and drives differential expression of key genes associated with NSCLC. DLG-369 can serve as a chemical probe to further interrogate the biological role of the GAS41 YEATS domain in NSCLC and other cancers for potential therapeutic implications. Citation Format: Alyssa Winkler, Dymytrii Listunov, Brian Linhares, Joshua Ray, Sidney Weaver, Sergey Zolov, Venkat Keshamouni, Jolanta Grembecka, Tomaszc Cierpicki. Targeting the GAS41 YEATS domain in non-small cell lung cancer with novel small molecules [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3239.

Human activities recognition from video images by using convolutional neural network

Nowadays, automatic human activity recognition from video images is necessary for monitoring applications and caring for disabled people. The use of surveillance cameras and the processing of the obtained images leads to the achievement of a smart, accurate system for the recognition of human behavior. Since human detection in different scenes is associated with many challenges, several approaches have been implemented to detect human activity from video image processing. Due to the complexity of human activities, background noises and other factors affect the detection. For the solution of these problems, two deep learning-based algorithms have been described in the current article. According to the convolutional neural networks, the LSTM + CNN method and the 3D CNN method have been used to recognize the human activities in the images of the video. Each algorithm is explained and analyzed in detail. The experiments designed in this paper are performed by two datasets: the HMDB-51 dataset and the UCF101 dataset. In the HMDB-51 dataset, the highest obtained accuracy for CNN + LSTM method was equal to 70.2 and for method 3D CNN equal to 54.4. In the UCF101 dataset, the highest obtained accuracy for CNN + LSTM method was equal to 95.1 and for method 3D CNN equal to 90.8.