Search Results (184)

Simulator-based digital twins are widely used in robotics education and industrial development to accelerate prototyping and enable safe experimentation. However, they often hide implementation details that are essential for understanding, diagnosing, and correcting system failures. This paper introduces a technology-independent model-based design framework that provides students with full visibility of the computational mechanisms underlying robotic controllers while remaining feasible within a 150-h undergraduate course. The approach relies on representing controller behavior using networks of Extended Finite State Machines (EFSMs) and their stacked extension (EFS2M), which unify all abstraction levels of the control architecture—from low-level reactive behaviors to high-level deliberation—under a single formal model. A structured programming template ensures traceable, optimization-free software synthesis, facilitating debugging and enabling self-diagnosis of design flaws. The framework includes real-time synchronized simulation, transparent switching between virtual and physical robots, and a smart data logger that captures meaningful events for model updating and error detection. Integrated into the Intelligent Robots course, the system supports topics such as kinematics, control, perception, and simultaneous localization and mapping (SLAM) while avoiding dependency on specific middleware such as Robot Operating System (ROS) 2. Over three academic years, students reported positive hands-on experiences, strong adaptability to diverse modeling approaches, and consistently high survey ratings reflecting the course’s overall quality. The proposed environment thus offers an effective methodology for teaching end-to-end robot controller design through transparent, simulation-driven digital twins. Full article

Background: Early childhood represents a key stage for the development of movement behaviors (MB), motor skills (MS), and executive functions (EF). Body Mass Index (BMI), defined according to World Health Organization (WHO) references, may influence these domains early in life. In this context, this cross-sectional observational study aimed to examine the associations between BMI and 24-h MB, MS, and EF in Tunisian preschool children aged 4 to 5 years. Methods: This cross-sectional observational study included 112 Tunisian children aged 4 to 5 years (50 boys, 62 girls), recruited from kindergartens in urban and rural areas. Anthropometric measurements were used to calculate age-specific BMI z-scores and classify children into three BMI categories: below normal, normal, and above normal. Twenty-four-hour MB physical activity (PA), sedentary behavior (SB), and sleep were objectively assessed using accelerometry over five consecutive days. EF (inhibition and working memory) were assessed using standardized cognitive tests, gross MS were evaluated using the Supine Timed Up and Go test (functional mobility), One-Leg Standing Balance test (postural steadiness), Hand Grip Dynamometer (upper body strength), and Standing Long Jump (lower body strength), and fine MS were assessed using the 9-Hole Pegboard Test (dexterity). All tools are validated and standardized for children. Results: Significant differences between BMI categories were observed for anthropometric variables (p < 0.05). In contrast, no significant differences were found for 24-h MB, adherence to recommendations, EF, and MS (p > 0.05). Only Sleep duration showed a difference significantly between BMI < normal and BMI > normal (p = 0.022). Conclusions: In Tunisian preschool children, weight status is primarily associated with differences in physical growth, with no marked relationship to MB, EF, or MS. These findings highlight the importance of universal preventive interventions, particularly focusing on growth monitoring, starting in early childhood. These results should be interpreted with caution and highlight the need for further studies on larger populations to better understand the relationships between BMI, PA, and development in young children. Full article

Background: Sarcopenia is defined by decreased muscle strength along with low muscle quantity or quality. The assessment of muscle strength may be performed by grip strength test or chair stand test (CST) and both of these tests are treated as equivalent tools for assessing muscle strength. Heart failure with preserved ejection fraction (HFpEF) contributes to the progression of sarcopenia, and it is left ventricular diastolic dysfunction (LVDd) which primarily leads to the development of HFpEF. The aim of this study was to examine the relationship of muscle strength with echocardiographic parameters of LVDd in patients with CKD and eGFR ≤ 29 mL/min/1.73 m² not treated with dialysis. Methods: The study samples consisted of 46 men with CKD stages G4–G5 not treated with dialysis: 23 participants with HGS < 27 kg and 23 individuals with HGS ≥ 27 kg. The assessment of muscle strength was provided by the hand grip strength (HGS) test and the five-times sit-to-stand test (FTSST). Transthoracic echocardiography was performed with the use of a convex probe in conjunction with a Logiq P6 ultrasound system. Results: In G4–G5 CKD patients, upper limb muscle strength did not correspond to lower limb muscle strength. Participants with prolonged FTSST had a lower mean value of septal e’ and higher mean E/e’ compared to individuals with correct both HGS and FTSST. Participants with correct HGS and prolonged FTSST had the lowest mean left ventricular ejection fraction (LVEF), as well as the lowest mean tricuspid annular plane systolic excursion (TAPSE). Conclusions: In G4–G5 CKD patients not treated with dialysis, HGS and FTSST are not equivalent and should not be used interchangeably. In this population, decreased muscle strength is associated with LVDd and FTSST is more sensitive than HGS in the prediction of LVDd. Low muscle strength is also associated with systolic function of the left and right ventricle in G4–G5 CKD patients not treated with dialysis. Full article

The unicellular ciliate Paramecium caudatum undergoes a developmental transition from asexual binary fission to sexual reproduction during its mature stage. This transition is triggered by mating interactions between cells of complementary mating types, leading to aggregate formation, mating pairs, and the meiotic division of micronuclei. Although calcium-driven EF-hand kinases have been implicated as mating type proteins, the spatiotemporal dynamics of calcium signaling during conjugation have not been comprehensively characterized. In this study, we established a behavioral assay to isolate committed cells from aggregates immediately after mating onset, and developed an experimental system to monitor intracellular calcium fluctuations specifically expressed in these cells. By combining Ca²⁺/EGTA buffering and microinjection approaches, we manipulated extracellular and intracellular calcium levels and confirmed the continuous requirement of calcium ions for conjugation-specific functions. Two significant findings emerged. First, we identified, for the first time, a calcium atlas covering the entire cell, with ascending centers localized in the anterior, oral apparatus, and posterior regions. The calcium/Indo-1-AM fluorescence peaked at 6 h after mating initiation and declined gradually, but persisted until conjugation was completed at ~48 h. Second, we demonstrated that distinct intracellular calcium thresholds are required for each stage of mating, including maintenance of mating activity, commitment of micronuclei to meiosis, and two-stepwise formation of mating pairs. These thresholds function as regulatory checkpoints that coordinate subcellular localization and stage synchronization. Collectively, our findings highlight calcium ions as pivotal regulators of conjugation in Paramecium and propose a novel framework, the Paramecium calcium atlas, for understanding the cellular and molecular mechanisms underlying sexual reproduction in ciliates. Full article

Background/Objectives: Heart failure with reduced ejection fraction (HFrEF) is associated with significant neuropsychological burden, including cognitive impairment and mood disturbances. While sacubitril/valsartan has demonstrated cardiovascular benefits, its effects on cognitive and emotional functioning remain underexplored, particularly in Middle Eastern populations. We aimed to evaluate the impact of sacubitril/valsartan on intellectual capacity, cognitive function and mood in patients with HFrEF using an idiographic study design. Methods: This study was conducted in adult patients with HFrEF selected to take sacubitril/valsartan to improve their clinical status. Participants were assessed at baseline and 3 months after treatment initiation using Al Khoudh Cognitive Test, PHQ-9 and Raven’s Progressive Colored Matrices. Results: Following three months of treatment, participants showed a statistically significant improvement in left ventricular ejection fraction (LVEF) (p = 0.043), depression severity (p = 0.025) and a non-significant trend toward improvement in abstract reasoning scores (p = 0.051). On the other hand, participants did not demonstrate significant improvements in the cognitive subdomains assessed by the Al Khoudh Test. Among these subdomains, the largest improvement was observed with verbal fluency (p = 0.057). Improvements in LVEF were not significantly associated with the changes in mood (p = 0.93), cognitive function (p = 0.34) or verbal fluency (p = 0.46). Conclusions: This study provides preliminary, hypothesis-generating evidence of potential short-term improvement in mood and reasoning scores in HFrEF patients treated with sacubitril/valsartan. Notably, these changes were not attributed to the observed improvements in cardiac function. These findings underscore the need for further investigation into the neurocognitive benefits of sacubitril/valsartan in larger and more diverse populations. Full article

Calmodulins (CaMs), which are important calcium-binding proteins, play critical roles in plant stress responses. However, limited information is available regarding the biological functions of CaMs under drought stress. In this study, we identified and isolated a CaM gene, ZmCaM2, from maize (Zea mays L.) in length and encodes a 184-amino acid protein containing four EF-hand domains capable of specifically binding calcium ions (Ca²⁺). Subcellular localization analysis revealed that ZmCaM2 is localized to the nucleus and membrane. Functional characterization indicated that ZmCaM2 negatively regulates drought tolerance in maize by increasing malondialdehyde (MDA) and reactive oxygen species (ROS) content while decreasing antioxidant enzyme activity, proline (Pro) content, abscisic acid (ABA) content and relative water content (RWC). Moreover, ZmCaM2 reduced maize sensitivity to ABA treatment, suggesting that ZmCaM2 negatively regulates the drought tolerance of maize by relying on the ABA pathway. These findings provide new insights into the functional role of ZmCaM2 and may facilitate the development of drought-resistant maize cultivars. Full article

Various intersection structures are utilized in city-wide traffic network infrastructure by local transportation authorities to handle the exponentially increasing traffic loads in developing countries. In this regard, numerous studies have considered the notable positive contribution of the modern roundabouts in intersection performance as a prominent method utilized widely in our contemporary world. Properly designed roundabouts are vital components of sustainable transportation planning, as they significantly influence traffic efficiency, safety, and environmental performance. Accurate estimation of roundabout capacity is essential to ensure that they can accommodate anticipated traffic volumes without causing congestion, thereby contributing to energy efficiency and reducing emissions. Moreover, sustainable roundabout design supports the development of safer and more inclusive transportation networks by improving accessibility for all road users, thus strengthening the overall sustainability of urban mobility. The SIDRA (version 8.0), a traffic simulation software, is frequently employed in performance analysis and determining the effects of possible outcomes of different scenarios of roundabouts in today’s world. On the other hand, driver behaviors are found to play a significant role in software performance during the analysis process of roundabout capacity and performance. Therefore, in order to optimize the environmental factor (EF) representing driver behaviors in the SIDRA software, a Differential Evolution Algorithm-Based Bi-Level Calibration Model (DEBCAM) was introduced. Observation data collected from eight different modern-structured roundabouts through drones were run into the SIDRA simulation software; the average delays obtained were employed to estimate optimum EF values through DEBCAM. Observed average delay values were taken into consideration with respect to the delay values obtained as a result of the SIDRA calibration by using the GEH statistics. GEH values indicate the consistency of vehicle delay data obtained via the DEBCAM with observed data. Acquired results clearly suggest that the SIDRA software needs to be calibrated so that it can represent drivers’ behaviors. After determination of the optimum values of the EF parameter for calibration of the SIDRA software, the regression analysis was conducted through the Partial Least Squares (PLS) method. As a result of the analysis, a capacity estimation model was developed, which displayed a significant conformity with the SIDRA capacity estimation results. Our findings suggested that the parameter requirement for the roundabout capacity estimation can be decreased by employing the appropriate EF value for the roundabout that needs to be analyzed. Full article

Background/Objectives: Spinal epidural lipomatosis (SEL) is increasingly recognized as a possible radiological indicator of Metabolic Syndrome (MS) and visceral adiposity. However, the precise relationship between visceral adiposity and the accumulation of epidural fat (EF) remains unclear. This study aimed to investigate the association between visceral adipose tissue (VAT) and EF thickness using quantitative MRI analyses. Methods: We retrospectively reviewed all MRI scans performed at our institution over a 7-month period, from May to November 2024. Two radiologists measured and recorded the VAT maximum antero-posterior diameter at the L3 level, EF maximum diameter at the L5-S1 level, spinal canal antero-posterior diameter at the L5-S1 level, and subcutaneous fat (SF) when included in the MRI images (at the L3 level) in all the MRI scans. Results: A cohort of 516 patients was collected (320 women and 196 men; mean age 57.31 ± 18.45 years old). In 508 patients (98.4%) SF and VAT were both measurable, while in 8 patients VAT only was assessable on MRI scans. Pearson correlation identified significant associations between EF and VAT thickness (correlation coefficient > 20%; p < 0.05). A linear regression model confirmed a significant, albeit modest, positive relationship between VAT and EF (R² = 5.4%). A multivariate regression model incorporating age, sex, spinal canal size, VAT, and SF improved the explanatory power (adjusted R² = 16.7%), with VAT, spinal canal diameter, and age emerging as significant predictors of EF (p < 0.001). Conclusions: Our study revealed in a large cohort of patients that EF and VAT are directly associated. On the other hand, SF resulted in not being associated with EF. These findings support the emerging concept that SEL can be a radiological phenotype of visceral obesity and, by extension, of MS. Integrating EF measurement into standard MRI interpretation may facilitate the early detection of SEL and offer additional insights into patients’ underlying metabolic profile. Full article

Background: The ice plant (Mesembryanthemum crystallinum L.) is a typical halophyte with remarkable stress resistance traits, including salinity and alkalinity tolerance. As a crucial signaling transduction pathway for plant responses to environmental stress, the CBL-CIPK signaling system plays a key role in regulating plant stress resistance. Methods: This study systematically analyzed the composition characteristics of the CBL and CIPK gene families across 24 plant species, including the ice plant, using comparative genomics approaches. Results: A total of 297 CBL and 561 CIPK gene family members were identified across the 24 species. Within the ice plant genome, 9 CBL and 18 CIPK genes were identified. Compared to model plants like Arabidopsis thaliana, the ice plant possesses a relatively higher number of CIPK genes, which may be related to its specific adaptation to saline–alkaline environments. Phylogenetic analysis indicated that the ice plant CBL and CIPK genes could be classified into three and four subfamilies, respectively. Expression analysis revealed that several genes (e.g., McCBL1, McCBL4, McCIPK1, McCIPK2) were significantly upregulated under salt stress, suggesting their important roles in the salt stress response. Notably, ice plant CBL and CIPK genes exhibit significant structural diversity. For instance, McCBL3 contains significantly more CDS regions than other members, while CIPK genes can be divided into two types: single-CDS type and multi-CDS type. This structural variation may be associated with functional divergence during the evolution of the gene family. Furthermore, three-dimensional (3D) structure prediction showed that CBL proteins primarily consist of EF-hand domains and α-helices, whereas CIPK proteins additionally contain β-sheet domains, implying that this structural difference may be related to their distinct regulatory mechanisms. Conclusions: This study provides an important theoretical basis for a deeper understanding of the molecular mechanisms underlying the CBL-CIPK signaling pathway in the saline–alkaline stress response of the ice plant. Full article

Genes involved in disease resistance are crucial for plant immune systems, yet their transcriptional regulatory mechanisms remain poorly understood. SRC4, a key member of the soybean mosaic virus resistance cluster (SRC), encodes a Ca²⁺-binding EF-hand domain and possesses antiviral activity, but its expression regulation is unclear. Here, we systematically analyzed 4085 soybean (Glycine max) transcriptome datasets and conducted SMV inoculation experiments to characterize SRC4 expression patterns. Cis-acting element analysis identified 12 regulatory elements in the SRC4 promoter, including salicylic acid (SA)-responsive elements. Furthermore, a Pro_SRC4::GUS reporter vector was constructed and functional analysis was performed in tobacco (Nicotiana benthamiana) and transgenic Arabidopsis thaliana. SRC4 exhibited significantly higher basal expression than typical resistance genes (R genes) and was induced by SMV infection, SA treatment, and Ca²⁺ supplementation, with peak expression at 2–5 h post-treatment (hpi). In transgenic tobacco overexpressing NahG, neither SMV nor Ca²⁺ could induce Pro_SRC4::GUS expression, demonstrating that SRC4 transcriptional regulation is mediated through SA signaling pathways. SRC4 showed predominant expression in roots and leaves and responded to temperature stress. Transgenic plants overexpressing SRC4 exhibited enhanced tolerance to both 12 °C and 37 °C temperature stress. This study elucidates the molecular mechanisms underlying SRC4 transcriptional regulation through Ca²⁺ and SA signaling pathways, revealing its dual role in both biotic and abiotic stress responses, especially in temperature stress. Full article

This study examined the spatial and seasonal variations of trihalomethanes (THMs) and estimated the health risks associated with THM exposure in drinking water through various pathways. Water samples were collected from 14 distribution districts connected to the Ulutan Distribution System (UDS) and the Süleyman Bey Distribution System (SDS), which supply drinking water to Zonguldak Province, Türkiye. THMs were measured using the USEPA 551 method. The median total trihalomethanes (TTHMs) ranged from 41 μg/L to 71 μg/L, which is below the Turkish drinking water standard of 100 μg/L. Chloroform (TCM) was the most common trihalomethane in all distribution networks in UDS and SDS. On the other hand, pre-ozonation oxidation after chlorination in SDS disinfection caused the contribution of brominated THMs (62%) to THM formation to be higher than that of TCM (38%). The study on cancer risk reveals that ingestion (96%) poses the greatest risk of the investigated pathways, followed by dermal contact (3.95%), while inhalation has been found to have a negligible effect. The highest and lowest median TTHMs occurred during winter and summer. The findings of the study show that the distribution areas of Kozlu, Ömerli, Topçalı, and Uzunçayır, for both genders, exhibit an unacceptable cancer risk level according to the criteria established by the USEPA (>10⁻⁴). Bromodichloromethane (BDCM) and chlorodibromomethane (DBCM) are the main contributors to cancer risk for males and females in UDS and SDS. The hazard index (HI) data indicated that the HI value remained below one for both UDS and SDS. Sensitivity analysis of THMs demonstrated that exposure frequency (EF) was the primary parameter contributing to the maximum potential impact on the total cancer risk exposure frequency (EF), followed by body weight (BW) and exposure duration (ED). Further, the results provide valuable information for health departments and water management authorities, enabling the formulation of more specific and efficient policies to minimise THM levels in drinking water distribution networks. Full article

Fasciola hepatica remains a global health and economic concern, and treatment still relies heavily on triclabendazole. At the parasite–host interface, F. hepatica calcium-binding proteins (FhCaBPs) have a unique EF-hand/DLC-like domain fusion found only in trematodes. This makes it a parasite-specific target for small compounds and vaccinations. To enable novel therapeutic strategies, we report the first elevated-resolution structure of a full-length FhCaBP4. The apo structure was determined at 1.93 Å resolution, revealing a homodimer architecture that integrates an N-terminal, calmodulin-like, EF-hand pair with a C-terminal dynein light chain (DLC)-like domain. Structure-guided in silico mutagenesis identified a flexible, 16-residue β4–β5 loop (LTGSYWMKFSHEPFMS) with an FSHEPF core that demonstrates greater energetic variability than its FhCaBP2 counterpart, likely explaining the distinct ligand-binding profiles of these paralogs. Molecular dynamics simulations and AlphaFold3 modeling suggest that EF-hand 2 acts as the primary calcium-binding site, with calcium coordination inducing partial rigidification and modest expansion of the protein structure. Microscale thermophoresis confirmed calcium as the major ligand, while calmodulin antagonists bound with lower affinity and praziquantel demonstrated no interaction. Thermal shift assays revealed calcium-dependent stabilization and a merger of biphasic unfolding transitions. These results suggest that FhCaBP4 functions as a calcium-responsive signaling hub, with an allosterically coupled EF-hand–DLC interface that could serve as a structurally tractable platform for drug targeting in trematodes. Full article

Skeletal muscle development is a critical economic trait in livestock, governed by myogenic satellite cell regulation. Integrins mediate mechanical anchorage to the ECM and enable ECM–intracellular signaling. CIB2, as an EF-hand-domain protein involved in mechanotransduction, shows significant developmental regulation in goat muscle. Although the role of CIB2 in skeletal muscle growth is poorly characterized, we observed pronounced developmental upregulation of IB2 in postnatal goat muscle. CIB2 expression increased >20-fold by postnatal day 90 (P90) compared to P1, sustaining elevation through P180 (p < 0.05). Functional investigations indicated that siRNA-mediated knockdown of CIB2 could inhibit myoblast proliferation by inducing S-phase arrest (p < 0.05) and downregulating the expression of CDK4/Cyclin D/E. Simultaneously, CIB2 interference treatment was found to decrease the proliferative activity of goat myogenic satellite cells, yet it significantly promoted differentiation by upregulating the expression of MyoD/MyoG/MyHC (p < 0.01). Mechanistically, CTCF was identified as a transcriptional repressor binding to an intragenic region of the CIB2 gene locus (ChIP enrichment: 2.3-fold, p < 0.05). Knockdown of CTCF induced upregulation of CIB2 (p < 0.05). RNA-seq analysis established CIB2 as a calcium signaling hub: its interference activated IL-17/TNF and complement cascades, while overexpression suppressed focal adhesion/ECM–receptor interactions and enriched neuroendocrine pathways. Collectively, this study identifies the CTCF-CIB2–integrin α7β1–PI3K/AKT axis as a novel molecular mechanism that regulates the balance of myogenic fate in goats. These findings offer promising targets for genomic selection and precision breeding strategies aimed at enhancing muscle productivity in ruminants. Full article

S100 proteins, a family of Ca²⁺-binding proteins, play numerous roles in cellular processes such as proliferation, differentiation, and apoptosis. Recent evidence has highlighted their critical involvement in neuroinflammation, a pathological hallmark of various neurodegenerative disorders including Alzheimer’s disease, multiple sclerosis, and Parkinson’s disease. Among these proteins, S100B and S100A8/A9 are particularly implicated in modulating inflammatory responses in the CNS. Acting as DAMPs, they interact with pattern recognition receptors like RAGE and TLRs, triggering pro-inflammatory signaling cascades and glial activation. While low concentrations of S100 proteins may support neuroprotective functions, increased levels are often associated with exacerbated inflammation and neuronal damage. This review explores the dualistic nature of S100 proteins in neuroinflammatory processes, their molecular interactions, and their potential as biomarkers and therapeutic targets in neurodegenerative disease management. Full article

Low-temperature stress severely limits plant growth and reduces agricultural productivity. Calmodulin-like (CML) proteins are crucial calcium sensors in plant cold responses. Transcriptome analysis of cold-stressed Saussurea involucrata identified seven differentially expressed CML genes. qRT-PCR confirmed that SiCML6 was strongly induced at 4 °C and −2 °C. Bioinformatics analysis showed that SiCML6 encodes a transmembrane protein containing an EF-hand domain. This protein carries a signal peptide and shows the closest phylogenetic relationship to Helianthus annuus CML3. Its promoter contains ABA, methyl jasmonate (MeJA), and cold-response elements. Arabidopsis plants overexpressing SiCML6 showed significantly higher survival rates at −2 °C than wild-type plants. Under freezing stress, SiCML6-overexpressing lines exhibited reduced malondialdehyde content, relative electrolyte leakage, and ROS accumulation (H₂O₂ and O₂⁻), along with increased proline, soluble sugars, soluble proteins, and total antioxidant capacity (T-AOC). SiCML6 elevated the expression of cold-responsive genes CBF3 and COR15a under normal conditions and further upregulated CBF1/2/3 and COR15a at 4 °C. Thus, low temperatures induced SiCML6 expression, which was potentially regulated by ABA/MeJA. SiCML6 enhances freezing tolerance by mitigating oxidative damage through boosted T-AOC and osmoprotectant accumulation while activating the CBF-COR signaling pathway. This gene is a novel target for improving crop cold resistance. Full article