Functional Approach Research Articles

Abstract B027: ISG15 modulate cancer cell mechanics and metastasis in breast cancer

Abstract Metastasis in breast cancer is a major challenge for prognosis and effective treatment, primarily because cancer cells can create an immune-suppressive microenvironment that supports their progression and spread. The physical properties of cancer cells significantly influence their metastatic potential and their interaction with the environment. Identifying targets that impact both the physical characteristics of cancer cells, and their microenvironment is crucial for tackling metastatic disease. Interferon-stimulated gene 15 (ISG15), a ubiquitin-like protein, is often overexpressed in breast cancer tissues and is associated with poor prognosis, higher tumor grade, and increased invasiveness. Despite these associations, the precise role of ISG15 and its post-translational modification, ISGylation, in cancer cell behavior and interactions with their microenvironment remains unclear. To explore this, we interrogated the biophysical role of ISG15 by using mouse modeling and in vitro loss of function approaches. We utilized whole- body ISG15-deficient mice and conducted a series of experiments to assess the impact of ISG15 on cancer cell properties and their surrounding environment. Our preliminary results indicate that cells lacking ISG15 exhibit altered physical traits, such as increased stiffness and size, which may influence their metastatic behavior. Additionally, targeting ISG15 or disrupting ISGylation enhances the susceptibility of cancer cells to immune cell-mediated destruction. In ISG15- deficient mice, we observed significantly reduced tumor growth and metastasis compared to wild-type controls. These findings suggest that ISG15 modulates critical aspects of cancer cell behavior and their interactions with the microenvironment, contributing to immune evasion and metastatic spread. Our research highlights ISG15 as a promising therapeutic target and provides a foundation for developing new strategies to prevent and treat metastatic cancer Citation Format: Mohamed Haloul, Priya Shah, Vinay Pai, Faruk Hossen, James James Lee, Ekrem Emrah Er. ISG15 modulate cancer cell mechanics and metastasis in breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr B027.

Differential functional change in olfactory bulb and olfactory eloquent areas in Parkinson’s disease

Abstract Olfactory dysfunction, or hyposmia, frequently occurs as a prodromal symptom and ongoing sign of Parkinson’s disease. Functional MRI is a powerful tool for studying functional changes in the olfactory brain regions in patients with Parkinson’s disease. However, existing studies show inconsistent results and no study has measured olfactory functional MRI abnormalities in the human olfactory bulb directly. This is mainly due to the well-known susceptibility artifacts in conventional functional MRI images that affect several key olfactory-eloquent brain regions, and especially the olfactory bulb. In this study, olfactory functional MRI was performed using a recently developed functional MRI approach that can minimize susceptibility artifacts and measure robust functional MRI signals in the human olfactory bulb during olfactory stimulation. Experiments were performed on high magnetic field (7T) in 24 early (< 5 years of parkinsonian symptoms) Parkinson’s disease patients and 31 matched healthy controls. Our data showed increased functional MRI signal changes (ΔS/S) in the olfactory bulb in patients with early Parkinson’s disease, which correlated with behavioral olfactory measures. Temporally, functional MRI signals in the olfactory bulb returned to the pre-stimulus state earlier after reaching peak amplitude in patients with early Parkinson’s disease, implicating a faster olfactory habituation effect. The piriform cortex showed reduced numbers of activated voxels in patients with early Parkinson’s disease, which correlated with behavioral olfactory assessment. Several secondary olfactory regions including the orbitofrontal cortex, temporal pole, and amygdala exhibited reduced numbers of activated voxels and increased functional MRI signal changes in patients with early Parkinson’s disease. Our data also showed that functional MRI results are highly dependent on voxel selection in the functional analysis. In summary, we demonstrate differential spatial and temporal characteristics of olfactory functional MRI signals between the primary and secondary olfactory regions in patients with early Parkinson’s disease. These results may assist the development of novel quantitative biomarkers (especially in the early stages of Parkinson’s disease) to track and predict disease progression, as well as potential treatment targets for early intervention.

High-grade B-cell lymphoma not otherwise specified, with diffuse large B-cell lymphoma gene expression signatures: Genomic analysis and potential therapeutics.

High-grade B-cell lymphoma not otherwise specified (HGBCL, NOS) has overlapping morphological and genetic features with diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma (BL), leading to uncertainty in its diagnosis and clinical management. Using functional genomic approaches, we previously characterized HGBCL and NOS, that demonstrate gene expression profiling (GEP), and genetic signatures similar to BL. Herein, we characterize distinct HGBCL, NOS, cohort (n = 55) in adults (n = 45) and in children (n = 10), and compared the GEP, genomic DNA copy number (CN), and mutational spectrum with de novo DLBCL (n = 85) and BL (n = 52). This subgroup, representing ~60% of HGBCL, NOS, lack gene-expression signature of BL and double hit/dark zone lymphoma, but express DLBCL like signatures and are characterized by either GCB- or ABC-like mRNA signatures and exhibit higher genomic complexity, similar to de novo DLBCL, and show alteration in genes regulating B-cell activation (CD79B, MYD88, PRDM1, TBLIXR1, CARD11), epigenome (KMT2D, TET2) and cell cycle transition (TP53, ASPM). However, recurrent mutations in genes often mutated in BL (DDX3X, GNA13, CCND3), but rare in DLBCL, are also present in HGBCL-NOS, highlighting genetic heterogeneity. Consistent with mutation spectrum, frequent genomic CN alterations in genes regulating B-cell activation (del-PRDM1, gain-BCL6, -REL, -STAT3) and cell cycle regulators (del-TP53, del-CDKN2A, del-RB1, gain-CCND3) were observed. Pediatric cases showed GCB-DLBCL-like mRNA signatures, but also featured hallmark mutations of pediatric BL. Frequent oncogenic PIM1 mutations were present in adult HGBCL, NOS. In vitro analyses with pharmacologic or genetic inhibition of PIM1 expression triggered B-cell activation and NF-κB-induced apoptosis, suggesting that PIM1 is a rational therapeutic target.

Distributed adaptive finite-time fault-tolerant cooperative control of heterogeneous multi-agent systems with saturation and disturbances

In this paper, the issue of distributed adaptive finite-time fault-tolerant cooperative control (FT-FTCC) problem is investigated for multiple unmanned aerial vehicles (UAVs) and unmanned ground vehicles (UGVs) with unknown parameter uncertainties, actuator faults, input saturation and external disturbances. Starting from the dynamic models of the UAVs and UGVs, an unified control model is presented. Then, a sliding-mode estimator is presented to estimate the position of the leader for the followers which only uses the information from neighbours. Next, a distributed adaptive FT-FTCC scheme, which can also deal with the uncertainties, actuator faults, input saturation and disturbances, is proposed by utilising disturbance observers and neural networks. Based on Lyapunov function approach, the tracking errors of all followers subject to the pre-defined desired positions are uniformly ultimately bounded. Finally, simulations are given to validate the efficiency of the developed FT-FTCC scheme.

A strategy for scaling the hardening behavior in finite element modelling of geometrically exact beams

AbstractA yield function in the stress resultant space of geometrically exact beams based on the elastoplastic cross-sectional warping problem has been proposed by Herrnböck et al. (Comput Mech, 67(3):723–742, 2021). This plasticity framework has been extended with a hardening tensor to model the kinematic hardening effects in Herrnböck et al. (Comput Mech, 71(1):1–24, 2022). While this framework provides scaling for the yield surface in ideal plasticity, scaling in hardening plasticity has not yet been explored. This paper focuses on the numeric modelling of hardening beams and beam assemblies at different geometric scales. Discretization effects from the introduction of plasticity into the geometrically exact beam model are demonstrated. Furthermore, the effects of scaling are explored, and a method to mitigate undesirable effects in order to achieve a size-agnostic formulation is proposed. Consistent geometric scaling is demonstrated for two alternative scaling approaches of the yield function.

A close look at children’s and adolescents’ arguments: combining a developmental, educational, and philosophical perspective

AbstractIt is broadly admitted that social contexts of reasoning may prompt children and adolescents to improve the quality of their reasoning. However, it is not clear how this quality may be assessed when it comes to arguments expressed within oral interactions in diverse settings (whole-class or small-group discussions) by students of different ages and cultural backgrounds. This study aims to offer a methodological contribution to the issue of oral argument assessment of children and adolescents, by looking at a large, annotated corpus of dialogic discussions during 111 lessons taking place in five countries. Our analysis combines a structural (Toulmin Argument Pattern) and functional (Walton’s argumentation schemes) approach to argumentative reasoning. Our findings show significant variations across age groups and social settings. The discussion points out the importance of sociocultural framing of argument reasoning development and the continuation of research in argument assessment methods able to grasp important developmental and cultural variations.

Creative music therapy in preterm infants effects cerebrovascular oxygenation and perfusion.

Creative music therapy (CMT) has been shown to promote the development of brain function and structure in preterm infants. We aimed to investigate the effect of CMT on cerebral oxygenation and perfusion to examine how the brain reacts to CMT. Absolute levels of cerebrovascular oxygen saturation (StO2) were measured in clinically stable preterm-born neonates (n = 20, gestational age: ≥30 weeks and < 37 weeks) using two near-infrared spectroscopy (NIRS)-based tissue oximeters over the right prefrontal cortex and left auditory cortex. We applied the systemic physiology augmented functional NIRS approach. Each CMT session lasted 55min and involved 9 intervals, including two 10-minute intervals during which the music therapist hummed and held the neonate. We found that CMT-induced changes in cerebrovascular StO2, perfusion and systemic physiology (i) could be classified into two groups (group 1: increase in StO2 during the first singing interval, group 2: decrease in StO2), (ii) differed in female neonates compared to male neonates, and (iii) correlated with individual blood haematocrit levels. Our exploratory study (i) demonstrates the impact of CMT on the neonate's physiology and (ii) highlights the need to analyze functional NIRS measurements in neonates separately according to their response pattern to avoid erroneous conclusions, e.g. when only the group average of the signal change is determined.

Africanized honeybee venom (Apis mellifera) promotes human complement activation split products storm

IntroductionComplement activation split products are signatures of many immunopathological disorders. Among the laboratory findings observed in these diseases, a reduction in the level of circulating intact complement components can be mentioned, and this change has also been detected in envenomation by multiple Africanized honeybee (Apis mellifera) stings. Although envenomation by these animals elicits diverse life-threatening reactions, the capacity of bee venom (AmV) to activate the human complement system remains elusive.Methods and findingsBy coupling immunochemical and functional approaches, it was observed that AmV strongly consumes components of the alternative pathway (AP) of the complement system in normal human serum (NHS). Additionally, AmV interfered with classical (CP) and lectin pathways (LP) activities. In parallel, a high increase in Ba fragment levels was detected, suggesting that the changes in AP activity were due to its activation. Furthermore, an increase in the level of the C1s-C1INH complex and a decrease in the physiological level of MASP1-C1INH suggested that CP and LP were also activated in the presence of AmV. Strikingly, NHS exposed to increasing AmV concentrations varying from 5 to 1000 µg/mL presented a high generation of C3a, C4a and C5a anaphylatoxins, and sC5b-9 complexes assembly, thus reinforcing that AmV triggers complement activation.ConclusionThese results show that AmV is a strong complement activator. This activation presents a mixed profile, with a predominance of AP activation. This suggests that complement split products can play important roles in the envenomation by Africanized honeybee, as they could induce diverse immunopathological events observed in patients and may also dictate patient clinical prognosis.

Read-write mechanisms of H2A ubiquitination by Polycomb repressive complex 1.

Epigenetic inheritance of silent chromatin domains is fundamental to cellular memory during embryogenesis, but it must overcome the dilution of repressive histone modifications during DNA replication1. One such modification, histone H2A lysine 119 monoubiquitination (H2AK119Ub), needs to be re-established by the Polycomb repressive complex 1 (PRC1) E3 ligase to restore the silent Polycomb domain2,3. However, the exact mechanism behind this restoration remains unknown. Here, combining cryo-electron microscopy (cryo-EM) and functional approaches, we characterize the read-write mechanism of the non-canonical PRC1-containing RYBP (ncPRC1RYBP). This mechanism, which functions as a positive-feedback loop in epigenetic regulation4,5, emphasizes the pivotal role of ncPRC1RYBP in restoring H2AK119Ub. We observe an asymmetrical binding of ncPRC1RYBP to H2AK119Ub nucleosomes, guided in part by the N-terminal zinc-finger domain of RYBP binding to residual H2AK119Ub on nascent chromatin. This recognition positions the RING domains of RING1B and BMI1 on the unmodified nucleosome side, enabling recruitment of the E2 enzyme to ubiquitinate H2AK119 within the same nucleosome (intra-nucleosome read-write) or across nucleosomes (inter-nucleosome read-write). Collectively, our findings provide key structural and mechanistic insights into the dynamic interplay of epigenetic regulation, highlighting the significance of ncPRC1RYBP in H2AK119Ub restoration to sustain repressive chromatin domains.

Semantic and distributional patterns of Spanish negation with nouns and adjectives: A Lexical-Realizational Functional Grammar approach

This study examines the semantic and distributional characteristics of the Spanish negation no when it occurs before nouns and adjectives, specifically, whether these instances diverge from sentential negation, but also from negation via prefixes such as des- ‘dis-’ or in- ‘in-, un-’. Using data from the Spanish Web 2011 corpus, it is shown that the use of no before adjectives aligns with other forms of constituent negation, often resulting in a contradictory interpretation, that is similar to that of sentential negation. However, the interpretation of no preceding nouns exhibits a broader range of interpretations, depending on whether the noun refers to an eventuality, quality, or entity. In such instances, no shows parallels to negative prefixes, frequently indicating privation rather than contradiction. Consequently, no can be analyzed as a building block of phrases, expressing syntactic negation, behaving similarly to other adverbs, as well as a building block of words, expressing lexical negation, resembling Spanish prefix-like elements like cuasi ‘almost’ or ex ‘ex-’. The analysis is couched within the framework of Lexical-Realizational Functional Grammar (LRFG), integrating aspects of Lexical Functional Grammar and Distributed Morphology. This approach does not treat morphology and syntax as separate modules, thus allowing us to account for the variable distribution and interpretation of no by means of a single representation in vocabulary structure, drawing on constituent structure representations which are needed on independent grounds for adverbs and for prefix-like elements.

Estimation of Cation Exchange Capacity for Low-Activity Clay Soil Fractions Using Experimental Data from South China

The cation exchange capacity (CEC) of the clay fraction (&lt;2 μm), denoted as CECclay, serves as a crucial indicator for identifying low-activity clay (LAC) soils and is an essential criterion in soil classification. Traditional methods of estimating CECclay, such as dividing the whole-soil CEC (CECsoil) by the clay content, can be problematic due to biases introduced by soil organic matter and different types of clay minerals. To address this issue, we introduced a soil pedotransfer functions (PTFs) approach to predict CECclay from CECsoil using experimental soil data. We conducted a study on 122 pedons in South China, focusing on highly weathered and strongly leached soils. Samples from the B horizon were used, and eight models and PTFs (four machine learning methods, multiple linear regression (MLR) and three PTFs from publication) were evaluated for their predictive performance. Four covariate datasets were combined based on available soil data and environmental variables and various parameters for machine learning techniques including an artificial neural network, a deep belief network, support vector regression and random forest were optimized. The results, based on 10-fold cross-validation, showed that the simple division of CECsoil by clay content led to significant overestimation of CECclay, with a mean error of 14.42 cmol(+) kg−1. MLR produced the most accurate predictions, with an R2 of 0.63–0.71 and root mean squared errors (RMSE) of 3.21–3.64 cmol(+) kg−1. The incorporation of environmental variables improved the accuracy by 2–10%. A linear model was fitted to enhance the current calculation method, resulting in the equation: CECclay = 15.31 + 15.90 × (CECsoil/Clay), with an R2 of 0.41 and RMSE of 4.48 cmol(+) kg−1. Therefore, given limited soil data, the MLR PTFs with explicit equations were recommended for predicting the CECclay of B horizons in humid subtropical regions.

Abstract A006: Development of a circulating tumor cell-derived cancer organoid platform for drug screening against metastatic tumors and longitudinal monitoring of drug resistance

Abstract Metastatic disease remains the primary cause of cancer-related deaths, presenting a significant challenge in oncology drug development. Current drug screening efforts often rely on cell lines or primary tumor-derived models, which may not adequately mimic the behavior of metastatic tumors. To address this, we aimed to develop a system that more accurately reflects metastatic tumors for improved drug screening. We leveraged a specialized culture platform featuring patented coated plates and proprietary media developed by AcroCyte (Taiwan) to cultivate circulating tumor cells (CTCs) into cancer organoids without pre-selection or enrichment. Building on our previous work, where we successfully cultivated CTCs from fresh blood and cryopreserved peripheral blood mononuclear cells (PBMCs) from renal, colon, and lung cancer patients into cancer organoids, we have now demonstrated the ability to freeze these CTC- derived organoids and thaw them for new culture when needed. We also present new data extending this success to breast and prostate cancers. CTCs from five frozen PBMC samples from breast cancer patients and three from prostate cancer patients formed visible colonies, with positive immunostaining for EpCAM, cytokeratin, and CD44. The immune cells in the PBMC samples gradually died off within three weeks of culture, as confirmed by the absence of CD45- positive cells. Interestingly, CTC-derived organoids from breast and prostate cancers did not form the spheroids observed in renal, colon, and lung cancer models. Instead, they appeared as colonies or clusters of CTCs. After 6-8 weeks of active growth, these clusters exhibited a slower growth rate, although they remained viable in the proprietary culture system. We are actively evaluating various culture media supplements, tailored to breast and prostate cancers, to extend the active growth period of these CTC-derived organoids for longer durations. Importantly, these CTC-derived colonies retained drug sensitivity, demonstrating the system's potential for effective drug screening against metastatic tumors. Our platform now supports organoid development from renal, colon, lung, breast, and prostate cancers, and we are actively expanding to include all major solid tumors. Since this technology uses blood, which can be obtained non-invasively, it enables longitudinal monitoring of CTCs as a functional liquid biopsy. Additionally, frozen PBMCs can be used as a source material for CTCs, and the resulting organoids can be utilized for drug sensitivity testing. This platform could be instrumental in oncology drug development and holds promise for monitoring emerging drug resistance in clinical trials by analyzing PBMC samples collected before and after treatment, offering a functional liquid biopsy-based approach to aid in developing therapies targeting metastatic disease. Citation Format: Yi-chun Han, David Hsieh, Ruobo Zhang, Emily Miyashita-Lin, Shian-Jiun Shih. Development of a circulating tumor cell-derived cancer organoid platform for drug screening against metastatic tumors and longitudinal monitoring of drug resistance [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A006.

The Functional Response of Estuarine Fish Communities to Hydrologic Change in a Semi-Arid Ecosystem

Functional assessment approaches were used to identify the responses of fish to environmental change in the San Antonio Bay System (Texas, USA). Using a 26-year coastal fisheries dataset (1993–2018), multivariate analyses revealed relationships between functional group abundance and freshwater inflows in the upper segments (Hynes Bay and Guadalupe Bay), but the patterns were decoupled from inflows in the lower bay segments (San Antonio Bay, Ayres Bay and Espiritu Santo Bay). In Hynes and Guadalupe Bays, freshwater migrant carnivores accounted for a significant fraction of the community irrespective of the gear, year or flow. Freshwater stragglers (omnivores and carnivores) were often present in the upper reaches of the bay. In the lower reaches, marine migrant omnivores were present during high and low flows in Espiritu Santos Bay, but only during low flows in Ayres Bay. Marine migrant carnivores were more important in gill nets irrespective of the flow conditions. The five most abundant fish were estuarine resident carnivores and omnivores, accounting for 53.5% of the community. Declines in the abundance of functional groups occurred during the 2011–2014 drought, with rebounds in 2015–2018. Functional methodologies provide insights into estuarine ecosystems and can serve as management tools to assess changes in fish assemblages.

Effects of regional network economies on industrial productivity in Japan: dynamic total factor productivity function approach

AbstractImproving productivity performance is essential for achieving regional economic sustainability. With the advent of the network society, regional economic agents have benefited not only from local within-region interactions but also from external out-of-region economies. However, it remains unclear whether localized external economies or spillovers of externalities through inter-regional networks are more significant in improving productivity performance. Therefore, this study examined the relationships between productivity performance and geographical externalities from an industry perspective using regional economic data for Japan and employed a dynamic total factor productivity function approach. It also assessed the impact of high-quality transportation infrastructure improvements on productivity catch-up. The results revealed the long-run effects of inter-regional networks on the productivity performance of industries. Furthermore, it revealed that, as a geographical externality, the “borrowed size” effect contributed significantly to productivity improvements in both manufacturing and non-manufacturing industries. These findings suggest that enhancing both high-quality transportation infrastructure and inter-regional transportation networks is likely to improve industry productivity performance.

Functional approaches in tendinopathy rehabilitation: exploring the role of tendon neuroplastic training

Functional approaches in tendinopathy rehabilitation: exploring the role of tendon neuroplastic training

Brain Injury and Neurodegeneration: Molecular, Functional, and Translational Approach 2.0

The brain is composed of different cells, such as neurons, glia, endothelial cells, etc [...]

In or outside the box? Citizen action between institutional and extra-institutional channels in Chinese environmental governance

ABSTRACT Authoritarian leaders around the world have recognized that citizen engagement is a core element of successful environmental governance. Faced with a massive environmental crisis, also authoritarian China is mobilizing its citizens to address its ecological problems. However, taking a functional approach to public participation, the Chinese party-state like other authoritarian regimes attempts to channel citizen action into institutional pathways and away from more contentious extra-institutional ones. Little research has investigated citizens’ reactions to such attempts. This paper analyses citizen participation between institutional and extra-institutional venues at the case of Chinese environmental governance. It finds that Chinese citizens use a mix of both channel types, but are overall more drawn to extra-institutional channels despite the risks. Moreover, our findings point to different motivations for the use of venues: personal grievances are more linked with the use of institutional channels, broader environmental concerns with extra-institutional ones. This shows, first, that channeling attempts have not yet been successful in crowding out noninstitutional channels. And second, that citizens seem to attach different meanings to the different channel types. This will make it hard for the Chinese and other authoritarian regimes to phase out extra-institutional action without severe losses for the effectiveness of environmental governance.

Features of the Use of the Indefinite Pronoun “one” in Russian and Kyrgyz Languages

Analyzes the functioning of the lexeme “one” in the sense of indefiniteness based on materials presented on the website of the Russian National Corpus and on literary works in Russian and Kyrgyz languages. The connection between the indefinite pronoun “one” and the indefinite articles of some Indo-European languages is established, as well as the conditions and reasons for the use of this pronoun in speech are identified. The relevance of the article is due to the fact that the category of definiteness/indefiniteness in different languages is currently being actively studied from a functional approach. This article differs from others in that it examines the indefinite pronoun “one” in the Russian language in comparison with the Kyrgyz language.

The Discourse Markers 'Olha' And 'Viu' And Their Pragmatic Functions In Language Use

Objective: The objective of this study is to investigate and describe the pragmatic contexts of the discourse markers "olha" and "viu" in vernacular language, identifying patterns of functioning through a functionalist analysis of these phenomena. Theoretical Framework: The main theoretical concepts guiding this research are rooted in a functional approach to language, based on a dynamic conception of language (Bybee, 2016), where use, discourse, and grammar interrelate in linguistic structuring. Additionally, we mapped what key authors (Risso, Silva, and Urbano, 2006; Brinton, 2017; and Traugott, 2022) in the field say about discourse markers, concluding that these elements are multifunctional; although optional from a syntactic perspective, they play an essential discourse-pragmatic role, considering their thetic nature of monitoring and regulating interaction. Method: We adopted a mixed methodology (Cunha Lacerda, 2016), which includes a qualitative and quantitative approach. The study design involves analyzing the phenomenon based on data samples drawn from three corpora: the Corpus of Cultured Portuguese of Vitória da Conquista (PCVC), the Corpus of Popular Portuguese of Vitória da Conquista (PPVC), and the Linguistic Corpus of Ibicoara. These corpora contain a total of 591,425 words from oral interviews following Labovian standards. For data analysis, we used the freeware AntConc, a corpus analysis tool required for linguistic studies, allowing for detailed searches and analyses. After data collection and quantification, we proceeded to an interpretative analysis of each occurrence found. Results and Discussion: Data collection revealed a total of 284 occurrences, of which 147 were for the marker "olha" and 137 for "viu." Pragmatically, these markers demonstrated multifunctionality through six distinct functions, namely: prefacing: delaying or deviating from the expected initial response by the interlocutor; argumentation: marking the speaker's position in presenting or defending a viewpoint and inviting the interlocutor to share it; reported speech: introducing a reported statement in another communicative context; interjection: a phatic function indicating the speaker's feelings; seeking discursive approval: the speaker's request for approval from the interlocutor to continue the discourse; and feedback: complementing the quest for discursive approval and expressing a clear orientation of the listener toward the speaker through attentive engagement with the interlocutor's speech. These data allow us to affirm that the markers are indeed multifunctional and, from a discursive perspective, essential for constructing meaning. Research Implications: We aim to significantly impact the field of linguistic studies from a functionalist perspective based on usage, as well as its discussion in the context of regular education. Teachers can use this study to enhance classroom discussions on issues related to the oral modality of the language, reflecting on aspects of linguistic (in)adequacy as well as paradigmatic analysis. This study also suggests that linguistic discussions should be based on a broad approach that considers extralinguistic aspects. Originality/Value: This study contributes to linguistic discussions that extend beyond syntactic structure relations. By considering discourse markers as thetic elements, we shed light on the discursive aspect of language and how discourse markers function multifunctionally to structure meaning. In this regard, we understand markers as essential elements in the oral modality of language based on their pragmatic functioning, while also hoping to contribute to reducing the stigma surrounding these elements due to a lack of a more in-depth approach in the prescriptive compendiums present in classrooms.

Crowd Flow Prediction: An Integrated Approach Using Dynamic Spatial–Temporal Adaptive Modeling for Pattern Flow Relationships

ABSTRACTPredicting crowd flows in smart cities poses a significant challenge for the intelligent transportation system (ITS). Traffic management and behavioral analysis are crucial and have garnered considerable attention from researchers. However, accurately and timely predicting crowd flow is difficult due to various complex factors, including dependencies on recent crowd flow and neighboring regions. Existing studies often focus on spatial–temporal dependencies but neglect to model the relationship between crowd flow in distant areas. In our study, we observe that the daily flow of each region remains relatively consistent, and certain regions, despite being far apart, exhibit similar flow patterns, indicating a strong correlation between them. In this paper, we proposed a novel Multiscale Adaptive Graph‐Gated Network (MSAGGN) model. The main components of MSAGGN can be divided into three major parts: (1) To capture the parallel periodic learning architecture through a layer‐wise gated mechanism, a layer‐wise functional approach is employed to modify gated mechanism, establishing parallel skip periodic connections to effectively manage temporal and external factor information at each time interval; (2) a graph convolutional‐based adaptive mechanism that effectively captures crowd flow traffic data by considering dynamic spatial–temporal correlations; and (3) we proposed a novel intelligent channel encoder (ICE). The task of this block is to capture citywide spatial–temporal correlation along external factors to preserve correlation for distant regions with external elements. To integrate spatio‐temporal flexibility, we introduce the adaptive transformation module. We assessed our model's performance by comparing it with previous state‐of‐the‐art models and conducting experiments using two real‐world datasets for evaluation.