Search Results (22)

Cellular senescence is a state of the durable cell cycle arrest of dysfunctional cells, which has been associated with the promotion of tumor cell reprogramming into a stem cell state. We previously reported that the mixed lineage kinase (MLK) inhibitor CEP-1347 promotes the differentiation of glioma stem cells (GSCs)—key contributors to glioblastoma recurrence and therapy resistance—into non-stem tumor cells. However, we also noted that CEP-1347–treated GSCs exhibited a morphological change suggestive of senescence. Therefore, we herein investigated whether CEP-1347 induces senescence in GSCs and, consequently, if senescent GSCs may be eliminated using senolytics. Cell death induced by CEP-1347 in combination with senolytic agents or with the knockdown of anti-apoptotic BCL2 family genes, as well as the effects of CEP-1347 on the expression of senescence markers and anti-apoptotic Bcl-2 family proteins, were examined. The results obtained showed that CEP-1347 induced senescence in GSCs accompanied by the increased expression of Bcl-xL. Among the panel of senolytic agents tested, navitoclax, a BH3 mimetic, efficiently induced cell death in GSCs when combined with CEP-1347 at concentrations clinically achievable in the brain. The knockdown of Bcl-xL resulted in more pronounced GSC death in combination with CEP-1347 than that of Bcl-2. These results suggest that combining CEP-1347 with the targeting of Bcl-xL, the expression of which increases with CEP-1347-induced senescence, is a rational approach to ensure the elimination of GSCs, thereby improving the outcomes of glioblastoma treatment. Full article

Bladder dysfunction is among the most drastic and quality-of-life-reducing conditions after spinal cord injury (SCI). Neuroinflammation in the lower urinary tract (LUT) after SCI could be a key driver of neurogenic bladder dysfunction and tissue fibrosis. Leukotrienes, a group of highly active lipid mediators, are potent inflammatory mediators. Here, we explored the potential of early montelukast (MLK) therapy, a cysteinyl leukotriene receptor 1 antagonist, on LUT function and structure four weeks after severe SCI in rats. Rats (strain Lewis, female, n = 50) received a permanent bladder catheter, followed by a complete T9 spinal cord transection. MLK was given daily, starting on day one post-injury. Bladder and locomotor function were regularly assessed. Bladder tissue was histologically and immunhistochemically analyzed. Post-SCI, MLK concentrations in plasma and cerebrospinal fluid were clinically relevant. MLK improved bladder functionality. MLK had no impact on smooth muscle alignment and uroepithelial integrity at this early SCI time point. This pilot study gave first insights into early, continuous oral MLK treatment with the first promising results of preserved LUT function and possible subsequent improved tissue integrity. Full article

In separate traditions in the HB, the Rephaim are presented either as a living group of gigantic warriors or as shadowy figures of the underworld of Sheol. They are referred to as the rp’um in earlier Amorite Ugaritic texts, in which their role and status are much debated. This paper offers a hypothesis that, first, tracks rp’um/Rephaim antecedent traditions from the Sumerian heroic and funerary practices adopted by the Amorites to the tradition of the rp’um of the Ugaritic literature, and then tracks them on to the HB, through the Amorite connection to Mlk/Molech, in two different regional traditions found in the HB. Literary analysis and cross-cultural evidence regarding the Amorites are used to demonstrate the plausibility of this hypothesis. This paper also puts forth that: the name Hammurapi is a reference to a funerary practice and is a titular name; rpi is employed in its more basic sense of meaning “to restore/mend”; rp’um, following Good, is the passive participle, “restored/healed ones”; and Deut 2:10–11 and the biblical King Og texts do not support the Israelites having encountered living Rephaim warriors. Tracking the heroic and death-culture traditions shows that the antecedents to the biblical Rephaim were likely originally heroic-age warriors who, upon death, were cared for and were appealed to through funerary rituals for some benefit. However, these Amorite traditions were not fully understood by the Israelites when they encountered them and appropriated aspects in their representation of the Rephaim. Full article

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a persistent inflammatory disorder of the upper airways, severely impacting quality of life. Dupilumab, targeting type 2 inflammatory pathways, is effective in managing severe, uncontrolled CRSwNP. However, the comparative accuracy of endoscopic scoring systems in monitoring therapeutic response to dupilumab remains unclear. This study compared the accuracy of the nasal polyp score (NPS) and the modified Lund–Kennedy endoscopic score (M-LKS) in assessing dupilumab response. Methods: A retrospective cohort analysis included 66 severe CRSwNP patients treated with dupilumab at Padua University. Endoscopic scores (NPS and M-LKS), patient-reported outcome mesures (PROMs), and clinician-reported outcome measures (CROMs), including peak nasal inspiratory flow (PNIF) and the Sniffin’ Sticks test, were evaluated at baseline and over 24 months. Results: Both NPS and M-LKS showed significant reductions over time (p < 0.001), significantly correlating with PNIF (p < 0.001). Given time and patient, PNIF emerged to be the only covariate related to endoscopic scores. No significant differences were observed between NPS and M-LKS regarding clinical outcome associations, suggesting equivalent accuracy. PNIF was identified as a critical predictor of endoscopic improvement, highlighting its clinical utility. These findings reinforce the role of standardized endoscopic metrics in assessing the efficacy of biologic therapies for CRSwNP. Full article

Laccase production was evaluated in 108 fungal isolates recovered from the eastern coast of Saudi Arabia, a critical element in environmental biodegradation and biotransformation. The most active isolate was identified as Curvularia lunata MLK46 (GenBank accession no. PQ100161). It exhibited maximal productivity at pH 6.5, 30 °C, and incubation for 5 d, with 1% sodium nitrate and 1% galactose as the preferred nitrogen and carbon sources, respectively. Productivity was enhanced by NaCl, CuSO₄, and FeCl₃ supplementation, with a maximum at 0.3 mM, 0.2 mM, and 61.7 mM concentrations, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the purified enzyme through diethylaminoethyl (DEAE)-Sepharose chromatography revealed a prominent band at 71.1 kDa with maximum activity at pH 6 and stability at pH 6–9. Furthermore, it was optimally active at 50 °C and thermally stable at 50–80 °C with a half-life time (T_1/2) of 333.7 min to 80.6 min, respectively. Its activity was also enhanced by many metallic ions, especially Fe³⁺ ions; however, it was inhibited by Hg²⁺ and Ag⁺ ions. The enzyme demonstrated significant degradation of specific substrates such as 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), guaiacol, o-dianisidine, and 2,6-dichlorophenol, with a kinetic efficiency constant which ranged from 40.95 mM⁻¹ s⁻¹ to 238.20 mM⁻¹ s⁻¹. UV spectrophotometry confirmed efficient oxidation peaks by electron transition against guaiacol (at 300 nm), o-dianisidine (at 480 nm), ABTS (at 420 nm), and 2,6-dichlorophenol (at 600 nm). The results collectively demonstrate the potential of laccase from C. lunata MLK46 as a promising agent for the effective biodegradation of several industrial pollutants under extreme conditions. Full article

Background/Objectives: Montelukast (MLK), a leukotriene receptor antagonist, has been associated with neuropsychiatric side effects. This study aimed to rationally modify MLK’s structure to reduce these risks by optimizing its interactions with dopamine D2 (DRD2) and serotonin 5-HT1A receptors using computational molecular simulation techniques. Methods: A library of MLK derivatives was designed and screened using structural similarity analysis, molecular docking, molecular dynamics (MD) simulations, MM/PBSA binding free energy calculations, and ADME-Tox predictions. Structural similarity analysis, based on Tanimoto coefficient fingerprinting, compared MLK derivatives to known neuropsychiatric drugs. Docking was performed to assess initial receptor binding, followed by 100 ns MD simulations to evaluate binding stability. MM/PBSA calculations quantified binding affinities, while ADME-Tox profiling predicted pharmacokinetic and toxicity risks. Results: Several MLK derivatives showed enhanced DRD2 and 5-HT1A binding. MLK_MOD-42 and MLK_MOD-43 emerged as the most promising candidates, exhibiting MM/PBSA binding free energies of −31.92 ± 2.54 kcal/mol and −27.37 ± 2.22 kcal/mol for DRD2 and −30.22 ± 2.29 kcal/mol and −28.19 ± 2.14 kcal/mol for 5-HT1A, respectively. Structural similarity analysis confirmed that these derivatives share key pharmacophoric features with atypical antipsychotics and anxiolytics. However, off-target interactions were not assessed, which may influence their overall safety profile. ADME-Tox analysis predicted improved oral bioavailability and lower neurotoxicity risks. Conclusions: MLK_MOD-42 and MLK_MOD-43 exhibit optimized receptor interactions and enhanced pharmacokinetics, suggesting potential neuropsychiatric applications. However, their safety and efficacy remain to be validated through in vitro and in vivo studies. Until such validation is performed, these derivatives should be considered as promising candidates with optimized receptor binding rather than confirmed safer alternatives. Full article

RAC3 encodes a small GTPase of the Rho family that plays a critical role in actin cytoskeleton remodeling and intracellular signaling regulation. Pathogenic variants in RAC3, all of which reported thus far affect conserved residues within its functional domains, have been linked to neurodevelopmental disorders characterized by diverse phenotypic features, including structural brain anomalies and facial dysmorphism (NEDBAF). Recently, a novel de novo RAC3 variant (NM_005052.3): c.196C>T, p.R66W was identified in a prenatal case with fetal akinesia deformation sequence (a spectrum of conditions that interfere with the fetus’s ability to move), and complex brain malformations featuring corpus callosum agenesis, diencephalosynapsis, kinked brainstem, and vermian hypoplasia. To investigate the mechanisms underlying the association between RAC3 deficiency and this unique, distinct clinical phenotype, we explored the pathophysiological significance of the p.R66W variant in brain development. Biochemical assays revealed a modest enhancement in intrinsic GDP/GTP exchange activity and an inhibitory effect on GTP hydrolysis. Transient expression studies in COS7 cells demonstrated that RAC3-R66W interacts with the downstream effectors PAK1, MLK2, and N-WASP but fails to activate SRF-, AP1-, and NFkB-mediated transcription. Additionally, overexpression of RAC3-R66W significantly impaired differentiation in primary cultured hippocampal neurons. Acute expression of RAC3-R66W in vivo by in utero electroporation resulted in impairments in cortical neuron migration and axonal elongation during corticogenesis. Collectively, these findings suggest that the p.R66W variant may function as an activated version in specific signaling pathways, leading to a distinctive and severe prenatal phenotype through variant-specific mechanisms. Full article

Machine learning (ML) methods, particularly Reinforcement Learning (RL), have gained widespread attention for optimizing traffic signal control in intelligent transportation systems. However, existing ML approaches often exhibit limitations in scalability and adaptability, particularly within large traffic networks. This paper introduces an innovative solution by integrating decentralized graph-based multi-agent reinforcement learning (DGMARL) with a Digital Twin to enhance traffic signal optimization, targeting the reduction of traffic congestion and network-wide fuel consumption associated with vehicle stops and stop delays. In this approach, DGMARL agents are employed to learn traffic state patterns and make informed decisions regarding traffic signal control. The integration with a Digital Twin module further facilitates this process by simulating and replicating the real-time asymmetric traffic behaviors of a complex traffic network. The evaluation of this proposed methodology utilized PTV-Vissim, a traffic simulation software, which also serves as the simulation engine for the Digital Twin. The study focused on the Martin Luther King (MLK) Smart Corridor in Chattanooga, Tennessee, USA, by considering symmetric and asymmetric road layouts and traffic conditions. Comparative analysis against an actuated signal control baseline approach revealed significant improvements. Experiment results demonstrate a remarkable 55.38% reduction in Eco_PI, a developed performance measure capturing the cumulative impact of stops and penalized stop delays on fuel consumption, over a 24 h scenario. In a PM-peak-hour scenario, the average reduction in Eco_PI reached 38.94%, indicating the substantial improvement achieved in optimizing traffic flow and reducing fuel consumption during high-demand periods. These findings underscore the effectiveness of the integrated DGMARL and Digital Twin approach in optimizing traffic signals, contributing to a more sustainable and efficient traffic management system. Full article

This study compared flavored kefir (KFR) and flavored milk (MLK) as a recovery drink in endurance master athletes. Using a randomized, placebo-controlled, non-blinded crossover design, 11 males and females completed three testing visits whilst acutely ingesting either KFR, MLK, or water as a placebo (PLA). KFR supplementation occurred for 14 days before the KFR-testing day, followed by a 3-week washout period. Testing visits consisted of an exhausting-exercise (EE) bout, a 4-h rest period where additional carbohydrate feeding was provided, and a treadmill 5 km time trial (TT). The Gastrointestinal Symptom Rating Scale (GSRS) survey was assessed at four timepoints. Blood was collected at baseline and after the TT and was analyzed for I-FABP levels. No significant difference (PLA: 33:39.1 ± 6:29.0 min, KFR: 33:41.1 ± 5:44.4 min, and MLK: 33:36.2 ± 6:40.5 min, p = 0.99) was found between the groups in TT performance. The KFR GSRS total score was significantly lower than the PLA after EE (p = 0.005). No differences in I-FABP were observed between conditions. In conclusion, acute KFR supplementation did not impact TT performance or I-FABP levels but may have reduced subjective GI symptoms surrounding exercise when compared to MLK or PLA. Full article

Inflammation is a critically important barrier found in innate immunity. However, severe and sustained inflammatory conditions are regarded as causes of many different serious diseases, such as cancer, atherosclerosis, and diabetes. Although numerous studies have addressed how inflammatory responses proceed and what kinds of proteins and cells are involved, the exact mechanism and protein components regulating inflammatory reactions are not fully understood. In this paper, to determine the regulatory role of mixed lineage kinase 3 (MLK3), which functions as mitogen-activated protein kinase kinase kinase (MAP3K) in cancer cells in inflammatory response to macrophages, we employed an overexpression strategy with MLK3 in HEK293 cells and used its inhibitor URMC-099 in lipopolysaccharide (LPS)-treated RAW264.7 cells. It was found that overexpressed MLK3 increased the mRNA expression of inflammatory genes (COX-2, IL-6, and TNF-α) via the activation of AP-1, according to a luciferase assay carried out with AP-1-Luc. Overexpression of MLK3 also induced phosphorylation of MAPKK (MEK1/2, MKK3/6, and MKK4/7), MAPK (ERK, p38, and JNK), and AP-1 subunits (c-Jun, c-Fos, and FRA-1). Phosphorylation of MLK3 was also observed in RAW264.7 cells stimulated by LPS, Pam3CSK, and poly(I:C). Finally, inhibition of MLK3 by URMC-099 reduced the expression of COX-2 and CCL-12, phosphorylation of c-Jun, luciferase activity mediated by AP-1, and phosphorylation of MAPK in LPS-treated RAW264.7 cells. Taken together, our findings strongly suggest that MLK3 plays a central role in controlling AP-1-mediated inflammatory responses in macrophages and that this enzyme can serve as a target molecule for treating AP-1-mediated inflammatory diseases. Full article

Previous studies on consumer yogurt preferences have mainly focused on added sugar, nutrient content, and health claims, leaving several knowledge gaps that should be filled through in-depth research. In this study, a more complete multi-attribute preference model was developed using the number of probiotic types, type of milk source, presence of edible gels (GEL), and usage of health food labels as the main yogurt attributes. A choice experiment (CE) was then conducted to investigate the relationship between multiple attribute preferences and willingness-to-pay (WTP). A total of 435 valid questionnaires were collected by the convenience sampling method. The results show that (1) respondents highly value the health food label (HEA), followed by the number of probiotic types (PRO); (2) the highest WTP in the conditional logit (CL) model was New Taiwan Dollar (NTD) (USD 10.5 for HEA, and the lowest was NTD 1.0 for 100% milk powder (MLK₂); (3) in the random-parameter logit (RPL) model, the highest WTP was NTD 14.6 for HEA, and the lowest was NTD 2.8 for GEL; (4) the most preferred attribute combination of yogurt was “8 or more probiotic types”, “a blend of raw milk and milk powder”, “the absence of edible gels”, “the presence of a health food label”, and “a price premium of NTD 6–10”; (5) married respondents with children were more willing to pay extra for yogurt products with a higher number of probiotic types and a health food label. The results may help the food industry understand and pay attention to consumer needs, which will, in turn, provide a reference for future product development and marketing strategies. Full article

Powdery mildew on barley, caused by the pathogen Blumeria graminis f. sp. hordei, occurs worldwide and can result in severe yield loss. Germplasm of barley, including landraces, commercial cultivars, wild relatives and breeding lines are stored in more than 200 institutions. There is a need for characterization of this germplasm in terms of resistance to biotic and abiotic stresses. This is necessary in order to use specific accessions in breeding programs. In the present study, 129 barley landraces originated from Turkey and provided by the ICARDA genebank were tested for resistance to powdery mildew. Seedling resistance tests after inoculation with 19 differentiated isolates of B. graminis f. sp. hordei were used to postulate the presence of resistance genes. From the 129 landraces studied, plants of 19 (14.7%) of them showed resistance to infection with powdery mildew. Based on preliminary tests from these 19 landraces, 25 resistant single plant lines were selected for testing with differential powdery mildew isolates. Seven lines were resistant to all 19 isolates used. However, only one line (5583-1-4) showed resistance scores of zero against all isolates used. It is likely that this line possesses unknown, but highly effective genes for resistance. In five resistant lines it was not possible to postulate the presence of specific resistance genes. In 19 lines the presence of the genes Mlp, Mlk, Mlh, Mlg, Ml(CP), Mlat, Mla3, Mla6, Mla7 and Mla22 were postulated. These new sources of highly effective powdery mildew resistance in barley landraces from Turkey could be successfully used in breeding programs. Full article

It was shown that mixed lineage kinase 1 (MLK1) regulates pancreatic cancer growth; however, its role in prostate cancer remains unclear. We showed that MLK1 is a tumor marker in prostate cancer by analyzing clinical gene expression data and identified a novel MLK1 inhibitor (NSC14465) from the compound library of the National Cancer Institute (NCI) using a MLK1 protein structure. The inhibitory effects of MLK1 were validated by an in vitro kinase assay and by monitoring phosphorylation signaling, and the anti-proliferation function was shown in several prostate and pancreatic cancer cell lines. We also demonstrated anti-tumor ability and prevention of cancer-related weight loss in a syngeneic orthotopic mouse model of pancreatic cancer that mimicked the tumor growth environment in the pancreas. Our results demonstrate that the MLK1 inhibitor is an anti-tumor agent for malignant prostate and pancreatic cancers. Full article

Background. SARS-CoV-2 infected patients are suggested to have a higher incidence of thrombotic events such as acute ischemic strokes (AIS). This study aimed at exploring vascular comorbidity patterns among SARS-CoV-2 infected patients with subsequent stroke. We also investigated whether the comorbidities and their frequencies under each subclass of TOAST criteria were similar to the AIS population studies prior to the pandemic. Methods. This is a report from the Multinational COVID-19 Stroke Study Group. We present an original dataset of SASR-CoV-2 infected patients who had a subsequent stroke recorded through our multicenter prospective study. In addition, we built a dataset of previously reported patients by conducting a systematic literature review. We demonstrated distinct subgroups by clinical risk scoring models and unsupervised machine learning algorithms, including hierarchical K-Means (ML-K) and Spectral clustering (ML-S). Results. This study included 323 AIS patients from 71 centers in 17 countries from the original dataset and 145 patients reported in the literature. The unsupervised clustering methods suggest a distinct cohort of patients (ML-K: 36% and ML-S: 42%) with no or few comorbidities. These patients were more than 6 years younger than other subgroups and more likely were men (ML-K: 59% and ML-S: 60%). The majority of patients in this subgroup suffered from an embolic-appearing stroke on imaging (ML-K: 83% and ML-S: 85%) and had about 50% risk of large vessel occlusions (ML-K: 50% and ML-S: 53%). In addition, there were two cohorts of patients with large-artery atherosclerosis (ML-K: 30% and ML-S: 43% of patients) and cardioembolic strokes (ML-K: 34% and ML-S: 15%) with consistent comorbidity and imaging patterns. Binominal logistic regression demonstrated that ischemic heart disease (odds ratio (OR), 4.9; 95% confidence interval (CI), 1.6–14.7), atrial fibrillation (OR, 14.0; 95% CI, 4.8–40.8), and active neoplasm (OR, 7.1; 95% CI, 1.4–36.2) were associated with cardioembolic stroke. Conclusions. Although a cohort of young and healthy men with cardioembolic and large vessel occlusions can be distinguished using both clinical sub-grouping and unsupervised clustering, stroke in other patients may be explained based on the existing comorbidities. Full article

Background: To investigate the acute effects of a capsaicin analogue supplement on 10 km time-trial performance and physiological responses in amateur athletes. Methods: Twenty-one participants (age = 29.3 ± 5.5 years, weight 74.2 ± 11.3 kg, height 176.0 ± 0.0 cm, fat mass 12.7 ± 3.8%, $\dot{\mathrm{V}}$O_2max 62.7 ± 8.4 mL·k⁻¹·min⁻¹), completed two randomized, double-blind trials: capsaicin analogue condition (Capsiate (CAP) = 24 mg) or a placebo (PLA) condition. The participants consumed two doses of 12 mg of CAP or PLA capsule 45 min before and immediately at the start of each trial. The time required to complete 10 km, lactate concentration, maximum heart rate (HR_peak), and rating of perceived exertion (RPE) were recorded. Results: The 10 km time-trial performance (CAP = 45.07 ± 6.41 min vs. PLA = 45.13 ± 6.73, p = 0.828) was not statistically significantly different between conditions. No statistically significant differences between conditions were detected for lactate concentration (p = 0.507), HR_peak (p = 0.897) and RPE (p = 0.517). Conclusion: Two doses of a 12 mg Capsaicin analogue supplement did not improve performance and physiological responses in a 10 km running time-trial in amateur athletes. Full article