Search Results (117)

Main roads are usually equipped with traffic flow monitoring devices in the road network to record the traffic flow data of the main roads in real time. Three complex scenarios, i.e., Y-junctions, multi-lane merging, and signalized intersections, are considered in this paper by developing a novel modeling system that leverages only historical main-road data to reconstruct branch-road volumes and identify pivotal time points where instantaneous observations enable robust inference of period-aggregate traffic volumes. Four mathematical models (I–IV) are built using the data given in appendix, with performance quantified via error metrics (RMSE, MAE, MAPE) and stability indices (perturbation sensitivity index, structure similarity score). Finally, the significant traffic flow change points are further identified by the PELT algorithm. Full article

Background: The blood–brain barrier (BBB) plays a critical role in protecting the central nervous system (CNS) but also limits drug delivery. Insufficient knowledge of how the CNS promotes the onset and maintenance of peripheral neuropathic pain limits therapeutic methods for the treatment of persistent neuropathic pain. Thus, this study aimed to evaluate the ability of a novel combination of Palmitoylethanolamide (PEA) and Equisetum arvense L. (Equisetum A.L.) to cross the BBB and modulate nociceptive pathways. Methods: Using a humanised in vitro BBB tri-culture model, the permeability, cytotoxicity, and integrity of the barrier were assessed after exposure to two different PEA forms, PEA ultramicronized (PEA-um) and PEA80mesh, Equisetum A.L., and a combination of the last two samples. The samples exhibited no cytotoxicity, maintained tight junction integrity, and efficiently crossed the blood–brain barrier (BBB), with the combination displaying the highest permeability. The eluate from the BBB model was then used to stimulate the co-culture of CCF-STTG1 astrocytes and SH-SY5Y neurons pre-treated with H₂O₂ 200 µM. Results: Treatment with the combination significantly increased cell viability (1.8-fold, p < 0.05), reduced oxidative stress (2.5-fold, p < 0.05), and decreased pro-inflammatory cytokines (TNFα, IL-1β) compared to single agents. Mechanistic analysis revealed modulation of key targets involved in pain pathways, including decreased FAAH and NAAA activity, increased levels of endocannabinoids (AEA and 2-AG), upregulation of CB2 receptor expression, enhanced PPARα activity, and reduced phosphorylation of PKA and TRPV1. Conclusions: These findings suggest that the combination of PEA and Equisetum A.L. effectively crosses the BBB and exerts combined anti-inflammatory and analgesic effects at the CNS level, suggesting a possible role in modulating neuroinflammatory and nociception responses. Full article

This study aimed to comprehensively investigate the impact of soybean β-conglycinin on broiler growth performance, nutrient utilization, allergic and inflammatory responses, intestinal barrier integrity, and cecal microbiota. A total of 168 newly hatched (1-day-old) Arbor Acres broilers with similar body weights were randomly divided into 6 treatment groups with 4 replicates of 7 broilers per replicate: the control group received a soybean-free basal diet, while the remaining five experimental groups were provided with diets supplemented with purified soybean β-conglycinin at the doses of 1%, 2%, 3%, 4%, and 5%. The results demonstrated that dietary 1–5% β-conglycinin supplementation significantly reduced the growth performance, nutrient utilization, and content of digestive enzymes in broilers (p < 0.05). Dietary 1–5% β-conglycinin supplementation also significantly increased the serum levels of histamine, β-conglycinin-specific IgY and IgM, TNF-α, and IL-6 and decreased IL-10 levels; the 3% group had the highest levels of histamine, TNF-α, and IL-6 and the lowest levels of IL-10 (p < 0.05). β-conglycinin supplementation significantly down-regulated the mRNA expression of tight junction proteins, MUC2 and IL-10, and up-regulated the expression of TNF-α and IL-6 in the small intestinal mucosa (p < 0.05). Furthermore, the Shannon and Simpson indices were significantly reduced by dietary 1–5% β-conglycinin administration (p < 0.05). The relative abundance of beneficial bacteria (Blautia, Lactobacillus, and Butyricoccus) was significantly decreased in all treatments (p < 0.05). Taken together, these findings suggest that β-conglycinin induces allergic and inflammatory responses, impairs intestinal barrier integrity, and alters the intestinal microbial balance, ultimately leading to reduced nutrient utilization and inhibited growth in broilers. Notably, our study demonstrated that dietary supplementation with 1% β-conglycinin already had various negative effects on broilers, and particularly supplemental 3% β-conglycinin induced serious allergic and inflammatory reactions. Therefore, in the present study, it is recommended that the inclusion level of β-conglycinin in broiler formula feed should not exceed 1%, i.e., the β-conglycinin content in the diet should not exceed 0.6% (converted from β-conglycinin purification purity). Full article

Extracellular vesicles (EVs) contain bioactive substances and mediate a multitude of physiological functions. EVs can be found in most body fluids and are particularly abundant in semen. EVs have the potential to become a biomarker for the quality of boar semen. In this study, EVs were isolated from the semen of relatively young (10 months of age, Y-EVs) and old (30 months of age, O-EVs) duroc boars using ultracentrifugation. The isolated EVs were characterized using a transmission electron microscope, nanoparticle tracking analysis, and Western blotting. MicroRNA (miRNA) profiles and metabolomes were analyzed using high-throughput sequencing and liquid chromatography–mass spectrometry, respectively. The median particle sizes of Y-EVs and O-EVs were 151.3 nm and 162.1 nm, respectively. miR-148a-3p, miR-10b, miR-21-5p, miR-10a-5p, let-7a, etc., were identified as highly enriched miRNAs in seminal EVs of boars. Comparative analysis revealed 41 differentially expressed miRNAs and 132 differential metabolites between Y-EVs and O-EVs. Notably, 18 miRNAs were upregulated in O-EVs, such as miR-339-5p, miR-125a, miR-423-3p, and miR-29c, which were mainly enriched in endocytosis, focal adhesion, and adherens junction. KEGG pathway analysis further indicated that differential metabolites were enriched in glycerophospholipid metabolism. These results provide an insight into the functional roles of seminal EVs. Full article

Purpose: Forme fruste choledochal cyst (FFCC) is a choledochal cyst with minimal or no dilatation of the extrahepatic bile duct (EHBD) and is usually associated with an anomalous pancreaticobiliary junction (APBJ). While sharing similar symptoms, inflammation, and malignant potential with classic biliary cysts, FFCC is often overlooked on ultrasound. This paper aims to present the experience of two tertiary pediatric centers in managing FFCC. Methods: In this retrospective study, the clinical data of pediatric patients treated for FFCC at two tertiary pediatric surgical centers between 1 January 2008 and 31 December 2023 were analyzed. The primary outcome was the clinical success of the surgical procedure, defined by the resolution of symptoms and the absence of major complications. Secondary outcomes included postoperative complications, type and duration of surgical procedures, and length of hospital stay. All patients underwent biliary reconstruction via either Roux-en-Y hepatico-jejunostomy or hepatico-duodenostomy. Clinical outcomes, including postoperative complications and patient follow-up, were evaluated. Results: Fourteen children (9 girls, 5 boys; aged 18 months to 12 years) underwent surgical treatment of FFCC. The mean age at surgery was 5.3 ± 3.8 years, and the mean diameter of the common bile duct was 7.9 ± 1.2 mm. Thirteen patients underwent Roux-en-Y hepatico-jejunostomy, and one underwent hepatico-duodenostomy. Over a mean follow-up period of 6.2 ± 3.6 years, no cholangitis or anastomotic stricture cases were observed. Two patients (14.3%) experienced minor wound infections managed conservatively. Conclusions: FFCC remains a diagnostic challenge due to its subtle imaging findings and non-specific clinical presentation. However, once identified, surgical excision with biliary reconstruction, most commonly via Roux-en-Y hepatico-jejunostomy, is a safe and effective treatment with excellent long-term outcomes. Given the potential for serious complications if left untreated, FFCC should be actively considered in pediatric patients with unexplained pancreatitis or biliary symptoms, even in the absence of overt ductal dilatation. Full article

Objective: Falciparum malaria is a major global health concern, affecting more than half of the world’s population and causing over half a million deaths annually. Red cell invasion is a crucial step in the parasite’s life cycle, where the parasite invade human erythrocytes to sustain infection and ensure survival. Two parasite proteins, Apical Membrane Antigen 1 (AMA-1) and Rhoptry Neck Protein 2 (RON2), are involved in tight junction formation, which is an essential step in parasite invasion of the red blood cell. Targeting the AMA-1 and RON2 interaction with inhibitors halts the formation of the tight junction, thereby preventing parasite invasion, which is detrimental to parasite survival. This study leverages machine learning (ML) to predict potential small molecule inhibitors of the AMA-1–RON2 interaction, providing putative antimalaria compounds for further chemotherapeutic exploration. Method: Data was retrieved from the PubChem database (AID 720542), comprising 364,447 inhibitors and non-inhibitors of the AMA-1–RON2 interaction. The data was processed by computing Morgan fingerprints and divided into training and testing with an 80:20 ratio, and the classes in the training data were balanced using the Synthetic Minority Oversampling Technique. Five ML models developed comprised Random Forest (RF), Gradient Boost Machines (GBMs), CatBoost (CB), AdaBoost (AB) and Support Vector Machine (SVM). The performances of the models were evaluated using accuracy, F1 score, and receiver operating characteristic—area under the curve (ROC-AUC) and validated using held-out data and a y-randomization test. An applicability domain analysis was carried out using the Tanimoto distance with a threshold set at 0.04 to ascertain the sample space where the models predict with confidence. Results: The GBMs model emerged as the best, achieving 89% accuracy and a ROC-AUC of 92%. CB and RF had accuracies of 88% and 87%, and ROC-AUC scores of 93% and 91%, respectively. Conclusions: Experimentally validated inhibitors of the AMA-1–RON2 interaction could serve as starting blocks for the next-generation antimalarial drugs. The models were deployed as a web-based application, known as PLASMOpred. Full article

Classical Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the MECP2 gene, resulting in a devastating phenotype associated with a lack of gene expression control. Mouse models lacking Mecp2 expression with an RTT-like phenotype have been developed to advance therapeutic alternatives. Environmental enrichment (EE) attenuates RTT symptoms in patients and mouse models. However, the mechanisms underlying the effects of EE on RTT have not been fully elucidated. We housed male hemizygous Mecp2-null (Mecp2^-/y) and wild-type mice in specially conditioned cages to enhance sensory, cognitive, social, and motor stimulation. EE attenuated the progression of the RTT phenotype by preserving neuronal cytoarchitecture and neural plasticity markers. Furthermore, EE ameliorated defects in neuromuscular junction organization and restored the motor deficit of Mecp2^-/y mice. Treatment with plasma from young WT mice was used to assess whether the increased activity could modify plasma components, mimicking the benefits of EE in Mecp2^-/y. Plasma treatment attenuated the RTT phenotype by improving neurological markers, suggesting that peripheral signals of mice with normal motor function have the potential to reactivate dormant neurodevelopment in RTT mice. These findings demonstrate how EE and treatment with young plasma ameliorate RTT-like phenotype in mice, opening new therapeutical approaches for RTT patients. Full article

The tumor microenvironment and healing wounds both contain extremely high concentrations of adenosine triphosphate (ATP) compared to normal tissue. The P2Y2 receptor, an ATP-activated purinergic receptor, is typically associated with pulmonary, endothelial, and neurological cell signaling. Here, we examine ATP-dependent signaling in breast epithelial cells and how it is altered in metastatic breast cancer. Using rapid imaging techniques, we show how ATP-activated P2Y2 signaling causes an increase in intracellular Ca²⁺ in non-tumorigenic breast epithelial cells, approximately 3-fold higher than their tumorigenic and metastatic counterparts. The non-tumorigenic cells respond to increased Ca²⁺ with actin polymerization and localization to the cell edges after phalloidin staining, while the metastatic cells remain unaffected. The increase in intracellular Ca²⁺ after ATP stimulation was blunted to control levels using a P2Y2 antagonist, which also prevented actin mobilization and significantly increased cell dissemination from spheroids in non-tumorigenic cells. Furthermore, the lack of Ca²⁺ changes and actin mobilization in metastatic breast cancer cells could be due to the reduced P2Y2 expression, which correlates with poorer overall survival in breast cancer patients. This study elucidates the rapid changes that occur after elevated intracellular Ca²⁺ in breast epithelial cells and how metastatic cancer cells have adapted to evade this cellular response. Full article

The objective of this study was to determine whether hydroxy-selenomethionine (OH-SeMet) exerts better protective effects on sows against heat stress than sodium selenite (SeNa) or seleno-yeast (SeY). A total of 60 sows (Landrace × Yorkshire) were randomly allocated into the three groups and fed a base diet supplemented with SeNa, SeY, or OH-SeMet at 0.3 mg Se/kg under a heat stress condition for a reproductive cycle. Compared to SeNa or SeY, OH-SeMet could more effectively sustain offspring growth performance, as evidenced by an increased number of live-born piglets, higher litter weight at day 21, and greater litter body weight gain from days 1 to 21. OH-SeMet was more effective in supporting endogenous redox systems, as shown by enhanced levels of TXNRD and GSH and reduced levels of GSSG in the serum of sows, improved T-AOC, TXNRD, and GSH alongside decreased MDA and GSSG in the serum of piglets, and heightened T-AOC in the jejunum of piglets. Furthermore, among the two tested organic Se sources, OH-SeMet was more effective than SeY in regulating immune responses compared to SeNa. OH-SeMet reduced inflammation-related markers CRP, HP, MAP, LPS, IL-1β, IL-6, and TNF-α, some or all of which were reduced in the serum of sows and their offspring. In addition, OH-SeMet also showed reduced glucose, TG, and NEFA levels, along with elevated insulin levels in the serum of sows. Correspondingly, among the two organic forms of Se, particularly those sows fed OH-SeMet showed better gut protection for the sows’ offspring, as indicated by a reduced crypt depth and increased villus height/crypt depth ratio in the duodenum, jejunum, and ileum than those fed SeNa. Specifically, compared to SeNa or SeY, OH-SeMet upregulated the expression of selenoproteins (GPX6, TXNRD3, GPX4, and SELENON), the tight junction protein (ZO-1), and host defense peptide gene (pBD1, pBD2, pBD3, NPG3, NPG4), along with downregulating levels of inflammation factor (IL-1β, IL-6 and TNF-α) and pro-apoptotic factor (P53) in the jejunum of piglets. Taken together, OH-SeMet more effectively mitigated the adverse effects induced by heat stress in sows and their offspring. Full article

The demand for faster and more efficient optical communication systems has driven significant advancements in integrated photonic technologies, with optical switches playing a pivotal role in high-speed, low-latency data transmission. In this work, we introduce a novel design for an adiabatic optical switch based on the thermo-optic effect using silicon-on-insulator (SOI) technology. The approach relies on slow optical signal evolution, minimizing power dissipation and addressing challenges of traditional optical switches. Machine learning (ML) techniques were employed to optimize waveguide designs, ensuring polarization-independent (PI) and single-mode (SM) conditions. The proposed design achieves low-loss and high-performance operation across a broad wavelength range (1500–1600 nm). We demonstrate the effectiveness of a Y-junction adiabatic switch, with a tapered waveguide structure, and further enhance its performance by employing thermo-optic effects in hydrogenated amorphous silicon (a-Si:H). Our simulations reveal high extinction ratios (ERs) exceeding 30 dB for TE mode and 15 dB for TM mode, alongside significant improvements in coupling efficiency and reduced insertion loss. This design offers a promising solution for integrating efficient, low-energy optical switches into large-scale photonic circuits, making it suitable for next-generation communication and high-performance computing systems. Full article

Methamphetamine (METH) is a powerful addictive psychostimulant that gives rise to severe abusers worldwide. While many studies have reported on the neurotoxicity of METH, blood–brain barrier (BBB) dysfunction has recently attracted attention as an essential target in METH-induced pathological changes in the brain. However, its mechanism has not been fully understood. We found that METH increased paracellular permeability and decreased vascular integrity through FITC–dextran and trans-endothelial electrical resistance (TEER) assay in primary human brain endothelial cells (HBMECs). Also, redistribution of tight junction proteins (zonula occluden-1 and claudin-5) and reorganization of F-actin cytoskeleton were observed in METH-exposed HBMECs. To determine the mechanism of METH-induced BBB disruption, the RhoA/ROCK signaling pathway was examined in METH-treated HBMECs. METH-activated RhoA, followed by an increase in the phosphorylation of downstream effectors, myosin light chain (MLC) and cofilin, occurs in HBMECs. Pretreatment with ROCK inhibitors Y-27632 and fasudil reduced the METH-induced increase in phosphorylation of MLC and cofilin, preventing METH-induced redistribution of junction proteins and F-actin cytoskeletal reorganization. Moreover, METH-induced BBB leakage was alleviated by ROCK inhibitors in vitro and in vivo. Taken together, these results suggest that METH induces BBB dysfunction by activating the RhoA/ROCK signaling pathway, which results in the redistribution of junction proteins via F-actin cytoskeletal reorganization. Full article

Cognitive impairment induced by chronic methamphetamine (METH) exposure exhibits similarities to neurodegenerative disorders and is associated with blood–brain barrier (BBB) dysfunction. However, the potential involvement of β-catenin in maintaining BBB integrity during METH exposure remains unexplored. In this study, Y-maze and novel object recognition tests were conducted to assess cognitive impairment in mice exposed chronically to methamphetamine for 2 and 4 weeks. Gd-DTPA and Evans blue leakage tests revealed disruption of the BBB in the hippocampus, while chronic METH exposure for 2 and 4 weeks significantly decreased β-catenin levels along with its transcriptionally regulated protein, claudin5. Additionally, various neural injury-related proteins, such as APP, Aβ1–42, p-tau (Thr181) and p-tau (Ser396), as well as neuroinflammation-related proteins, such as IL-6, IL-1β, and TNF-α, exhibited increased levels following chronic METH exposure. Furthermore, plasma analysis indicated elevated levels of p-Tau (total), neurofilament light chain, and GFAP. In vitro experiments demonstrated that exposure to METH resulted in dose-dependent and time-dependent reductions in cellular activity and connectivity of bEnd.3 and hcmec/D3 cells. Furthermore, β-catenin exhibited decreased levels and altered subcellular localization, transitioning from the cell membrane to the cytoplasm and nucleus upon METH exposure. Overexpression of β-catenin was found to alleviate endothelial toxicity and attenuate junctional weakening induced by METH. The aforementioned findings underscore the crucial involvement of β-catenin in endothelial cells during chronic METH exposure-induced disruption of the BBB, thereby presenting a potential novel target for addressing METH-associated cerebrovascular dysfunction and cognitive impairment. Full article

Background: Patients with proximal junctional kyphosis (PJK) or failure (PJF) may demonstrate disparate outcomes and recovery when fused to the upper (UT) versus lower (LT) thoracic spine. Few studies have distinguished the reoperation and recovery abilities of patients with PJK or PJF when fused to the upper (UT) versus lower (LT) thoracic spine. Methods: Adult spine deformity patients ≥ 18 yrs with preoperative and 5-year (5Y) data fused to the sacrum/pelvis were included. The rates of PJK, PJK revision, and radiographic PJF were compared between patients with upper instrumented vertebra (UIV) in the upper thoracic spine (UT; T1-T7) and lower thoracic spine (LT; T8-L1). Mean differences were assessed via analyses of covariance, factoring in any differences between cohorts at baseline and any use of PJF prophylaxis. Backstep logistic regressions assessed predictors of achieving Smith et al.’s Best Clinical Outcomes (BCOs) and complications, controlling for similar covariates. Results: A total of 232 ASD patients were included (64.2 ± 10.2 years, 78% female); 36.3% were UT and 63.7% were LT. Postoperatively, the rates of PJK for UT were lower than LT at 1Y (34.6 vs. 50.4%, p = 0.024), 2Y (29.5 vs. 49.6% (p = 0.003), and 5Y (48.7 vs. 62.8%, p = 0.048), with comparable rates of PJF. In total, 4.0% of UT patients underwent subsequent reoperation, compared to 13.0% of LT patients (p = 0.025). A total of 6.0% of patients had recurrent PJK, and 3.9% had recurrent PJF (both p > 0.05). After reoperation, UT patients reported higher rates of improvement in the minimum clinically important difference for ODI by 2Y (p = 0.007) and last follow-up (p < 0.001). While adjusted regression revealed that, for UT patients, the minimization of construct extension was predictive of achieving BCOs by last follow-up (model p < 0.001), no such relationship was identified in LT patients. Conclusions: Patients initially fused to the lower thoracic spine demonstrate an increased incidence of PJK and lower rates of disability improvement, but are at a lessened risk of neurologic complications if reoperation is required. Full article

Pyroptosis is a gasdermin-mediated pro-inflammatory programmed cell death that, during microbial infections, aims to restrict the spreading of bacteria. Nevertheless, excessive pyroptosis activation leads to inflammation levels that are detrimental to the host. Pathogen-associated molecular patterns (PAMPs) present in bacteria and outer membrane vesicles (OMVs) can trigger pyroptosis pathways in different cell types with different outcomes. Moreover, some pathogens have evolved virulence factors that directly interfere with pyroptosis pathways, like Yersinia pestis YopM and Shigella flexneri IpaH7.8. Other virulence factors, such as those of Neisseria meningitidis, Neisseria gonorrhoeae, Salmonella enterica, and Helicobacter pylori affect pyroptosis pathways indirectly with important differences between pathogenic and commensal species of the same family. These pathogens deserve special attention because of the increasing antimicrobial resistance of S. flexneri and N. gonorrhoeae, the high prevalence of S. enterica and H. pylori, and the life-threatening diseases caused by N. meningitidis and Y. pestis. While inflammation due to macrophage pyroptosis has been extensively addressed, the effects of activation of pyroptosis pathways on modulation of cell cytoskeleton and cell–cell junctions in epithelia and endothelia and on the bacterial crossing of epithelial and endothelial barriers have only been partly investigated. Another important point is the diverse consequences of pyroptosis pathways on calcium influx, like activation of calcium-dependent enzymes and mitochondria dysregulation. This review will discuss the pyroptotic pathways activated by Gram-negative bacteria and their OMVs, analyzing the differences between pathogens and commensal bacteria. Particular attention will also be paid to the experimental models adopted and the main results obtained in the different models. Finally, strategies adopted by pathogens to modulate these pathways will be discussed with a perspective on the use of pyroptosis inhibitors as adjuvants in the treatment of infections. Full article

Due to its turbulent demographic history, marked by extensive settlement and gene flow from diverse regions of Eurasia, Southeastern Europe (SEE) has consistently served as a genetic crossroads between East and West and a junction for the migrations that reshaped Europe’s population. SEE, including modern Croatian territory, was a crucial passage from the Near East and even more distant regions and human populations in this region, as almost any other European population represents a remarkable genetic mixture. Modern humans have continuously occupied this region since the Upper Paleolithic era, and different (pre)historical events have left a distinctive genetic signature on the historical narrative of this region. Our views of its history have been mostly renewed in the last few decades by extraordinary data obtained from Y-chromosome studies. In recent times, the international research community, bringing together geneticists and archaeologists, has steadily released a growing number of ancient genomes from this region, shedding more light on its complex past population dynamics and shaping the genetic pool in Croatia and this part of Europe. Full article