Bioorthogonal reactions offer a powerful tool for site-specific labeling of biomolecules in living systems. Among them, the recently reported hydrazonyl sultone (HS)-bicyclo[6.1.0]non-4-yne (BCN) ligation reaction stands out for its fast reaction kinetics and tunable aqueous stability. Herein, we describe the experimental protocols of using HS-BCN ligation for site-specific modification of a recombinant nanobody in vitro and a G protein-coupled receptor (GPCR) on a live mammalian cell surface. These protocols include the genetic encoding of BCN-lysine (BCNK) into the target protein, bioorthogonal modification of the BCNK-encoded proteins, and characterization of the reaction rate and selectivity. Together with the robust genetic encoding of the strained alkyne BCN in any protein structure, the HS-BCN ligation reaction promises to expand the capabilities of bioorthogonal chemistry to enable facile modifications of domain antibodies in vitro for diagnostic applications and selective fluorescent labeling of GPCRs for biophysical studies of receptor dynamics in live cells.
Anaphylaxis attributable to non-immunoglobulin E (IgE)-mediated mechanisms represents an increasingly recognized and clinically challenging subset of severe hypersensitivity reactions. This review synthesizes recent advances in the understanding of IgE-independent pathways, with emphasis on literature published in the last 18 months, and highlights their implications for precision diagnostics and therapeutic targeting. Major IgE-independent mechanisms have attracted substantial recent attention. First, the Mas-related G protein-coupled receptor X2 (MRGPRX2) has been consolidated as a central mediator of IgE-independent drug reactions, with new humanized knock-in mouse models revealing its capacity to amplify both IgE-dependent and IgE-independent systemic anaphylaxis. Second, clonal mast cell disorders-including systemic mastocytosis and monoclonal mast cell activation syndrome-are now recognized as major risk amplifiers for severe and fatal anaphylaxis, particularly following Hymenoptera venom exposure, reinforcing the role of the KIT D816V mutation in lowering the mast cell activation threshold. Third, a comprehensive biomarker meta-analysis confirms that tryptase, platelet-activating factor (PAF), and urinary prostaglandin D2 each contribute differentially to non-IgE reactions, with PAF and PAF-acetylhydrolase emerging as particularly relevant in IgE-independent severity pathways. Non-IgE-mediated anaphylaxis encompasses mechanistically distinct entities that demand tailored diagnostic algorithms. Recognizing MRGPRX2 activation, complement and IgG-dependent pathways, and clonal mast cell disease in the clinical work-up of unexplained or recurrent anaphylaxis is relevant. Emerging biomarkers and therapeutic targets - including MRGPRX2 antagonists - hold promise for transforming the management of this underdiagnosed condition.
Osteosarcoma (OS) is a malignant neoplasm arising from bone tissue. The OS exhibited a significant degree of heterogeneity, with a high incidence of local invasion and a high rate of metastasis. Approximately 25% of those diagnosed with OS present with metastases, with the lungs being the most prevalent site, followed by the bones or lymph nodes. The 5-year survival rate for patients with OS with lung metastasis is only 10% to 30%. Adjuvant chemotherapy regimens significantly enhanced the 5-year survival rate in patients diagnosed with non-metastatic OS. However, the same regimens did not provide a substantial improvement in survival for patients with OS who had acquired lung metastasis or had recurrence. Moreover, the extensive use of chemotherapy medications often results in the development of resistance to chemotherapy in patients with OS, which frequently culminates in treatment ineffectiveness. Natural products (NPs) are significant reservoirs of anti-cancer medications. Preclinical investigations have shown that certain NPs had substantial promise for treating OS. However, there is currently a lack of a comprehensive overview of the use of NPs in OS treatment. In this review, we provide a comprehensive overview of the NPs, which include polyphenols, alkaloids, and terpenoids, that have the potential to be effective in the treatment of OS.
Proportional-integral-derivative (PID) controllers remain the cornerstone of industrial automation owing to their robustness and operational simplicity. However, tuning PID parameters for nonlinear processes presents persistent challenges, often requiring system linearization and repeated recalibration under varying operating conditions. Although both traditional and reinforcement learning (RL)-based auto-tuning methods have shown considerable promise, their dependence on direct trial-and-error interactions with live processes raises substantial safety and feasibility concerns in industrial environments. This study presents a novel PID generator framework that leverages virtual environments to enable safe and efficient offline training. Simplified first-order plus dead-time systems are constructed to emulate the slow nonlinear dynamics of target processes, serving as interactive surrogate environments for RL agent training. To address performance discrepancies under varying operating conditions, a reward normalization mechanism-a critical yet previously underexplored component for reliable RL-based PID tuning-is proposed. Furthermore, an enhanced actor-critic architecture is adopted to further improve learning efficiency and policy convergence. Once training is complete, the agent autonomously generates optimal PID parameters for target processes without requiring online retraining or direct process interaction. The effectiveness and robustness of the proposed framework are validated through comprehensive numerical simulations and real-world industrial experiments. The results demonstrate its capacity to adaptively and reliably determine optimal PID parameters for nonlinear systems, thereby bridging the gap between advanced RL methodologies and practical industrial control applications.
Here, we describe the covalent modification of cell surfaces with antibodies through bioconjugation to the self-labeling SNAP-tag enzyme. We apply this approach to the reprogramming of T cell signaling through the universal chimeric antigen receptor (CAR), "SNAP-CAR." Universal CARs are a highly programmable class of engineered receptors that interact with co-administered "adaptor" antibodies to recognize one or more antigens of interest to trigger receptor signaling. Universal CARs have promise for use in cell therapeutics and as research tools. SNAP-CAR covalently attaches to adaptor antibodies containing a benzyl guanine (BG) motif. The protocols presented here include methods for SNAP-CAR T cell and antibody adaptor generation using gamma retroviral transduction and BG-NHS ester conjugation, respectively. The chapter also describes methods to evaluate cell surface bioconjugate formation, including quantification of BG motifs on antibodies and co-incubation experiments to assay for antigen-directed SNAP-CAR T cell functions of target cell killing and SNAP-CAR T cell activation.
High ventilation breathwork (HVB) is increasingly used in psychotherapeutic and contemplative contexts to induce altered states of consciousness (ASC), yet its neurobiological mechanisms remain poorly understood. Thirty experienced HVB practitioners (mean age 44.97 ± 11.80 years) underwent perfusion and resting-state fMRI to assess acute cerebrovascular and post-acute network changes. Cerebral blood flow (CBF) was measured using arterial spin labelling during baseline and the interval of maximal subjective effects. Acute psychotropic effects were rated using an online Likert scale, ASC intensity was assessed immediately after breathwork with the 5-Dimensional Altered States of Consciousness (5D-ASC) scale. HVB induced a 45.5% reduction in global grey matter cerebral blood flow (CBF; 29.81 ± 7.58 vs. 16.24 ± 4.35 ml/100 g/min; t(29) = -10.69, p < 0.001), accompanied by increased heart and respiratory rate. Default mode network (DMN) functional connectivity (FC) did not change significantly at the group level. In further exploratory analyses, greater decreases in DMN FC (pre - post breathwork) predicted higher 5D-ASC rating (β = -4.88, p < 0.05) in analyses using global signal regression, though this association was not significant without global signal regression (p = 0.75). CBF reductions were not associated with ΔDMN-FC or 5D-ASC ratings. HVB produces marked global cerebral hypoperfusion without systematic DMN FC changes, suggesting that hypoperfusion and network dynamics may represent partially dissociable mechanisms underlying breathwork-induced ASC. As a controllable, non-pharmacological means of inducing ASC, HVB may hold promise as a psychotherapeutic and psychedelic-adjacent intervention.
To evaluate which imaging biomarkers in uveitis have approached clinical utility, which remain incompletely validated, and how consensus frameworks and artificial intelligence are reshaping the path toward standardized, decision-useful assessment. Quantitative optical coherence tomography (OCT) and optical coherence tomography angiography (OCTA) metrics have gained disease-specific precision. Anterior segment OCT particle-size analysis differentiates inflammatory cells from pigment with high discriminative performance. OCTA vessel density tracks chronic microvascular loss in birdshot chorioretinopathy more reliably than it captures real-time inflammatory activity. Choroidal vascularity index shows promise as a subclinical choroidal marker, though cross-platform standardization remains unresolved. The multimodal uveitis (MUV) international taskforce has established consensus minimal imaging sets for five white dot syndromes, providing the first systematic effort to define which modality is essential for diagnosis, activity monitoring, and complication detection in specific entities. Artificial intelligence applications have demonstrated proof-of-concept performance, but translation is constrained by small datasets and absent prospective validation. Most proposed imaging biomarkers remain disease-specific and incompletely validated. Recent quantitative approaches and international consensus frameworks may narrow the gap between descriptive imaging and actionable, trial-ready biomarkers. The bottleneck has shifted from image acquisition to validation, harmonization, and integration with clinical outcomes.
The growing availability of public genomic repositories led to increased use of integrative and meta-analytic approaches to combine multi-omics datasets for biomarker discovery. Building on this framework, our study applies a molecular discovery approach to identify diagnostic biomarkers for cervical pre-cancer in high-risk HPV-positive women using aggregated genomic evidence across populations. This study performed a secondary analysis of publicly available DNA-methylation datasets identified through a systematic search of the Gene Expression Omnibus database to enable integrative analysis. After employing standard preprocessing methods across the selected DNA-methylation datasets, statistical analyses (specifically limma and meta-analysis with a random-effects model) were used to calculate the cumulative standardized effect sizes for all 9570 genes across the 4 selected studies. This cross-dataset analysis led to the identification of 4 promising, non-population-specific genes, that show potential for detecting precancerous cervical lesions in high-risk HPV-positive women. Collectively, these genes reflect a molecular profile associated with Cervical Intraepithelial Neoplasia lesions undergoing malignant progression. By leveraging aggregated data, our findings enhance the potential of generalization of biomarker signals and illustrate the strength of multi-data sets integration in advancing cervical pre-cancer triage screening strategies. Given their diagnostic promise, further experimental validation in large and diverse cohorts is warranted to evaluate their clinical applicability.
Introduction In recent years, artificial intelligence (AI)-generated prostate whole-gland segmentation has shown promise for clinical use. This study compares AI- and human-derived prostate segmentation on magnetic resonance imaging (MRI) and evaluates its practical application. Methods A retrospective study from December 2020 to December 2022 evaluated 31 randomly selected patients who had previously undergone MRI-ultrasound (US)-guided fusion biopsies. AI-generated auto-contours of the whole-gland prostate, seminal vesicle, and urethra were produced using the ProtégéAI feature of the MIM Software from MRIs obtained on GE Signa HDxt 3.0T and Siemens Altea Magnetom 1.5T scanners. The same MRIs were independently contoured manually by a board-certified urologist (U) and a board-certified radiologist (R) using MIM Software contouring tools. Volumetric conformity among the three contour sets was assessed using the Dice similarity coefficient, Hausdorff distance (HD), and mean distance to agreement (MDA). Contouring time was recorded for each method, with AI timed from command execution to file generation and physicians timed from file opening to save. Pairwise Wilcoxon signed-rank tests (JMP Pro 15) compared contouring times and similarities across AI to urologist (AI-U), AI to radiologist (AI-R), and urologist to radiologist (U-R) contours. Results The average volumetric Dice similarity, HD, and MDA for the AI-U contours were 0.875±0.039, 9.186±4.004 mm, and 1.410±0.511 mm for the whole-gland prostate, 0.283±0.185, 18.117±13.265 mm, and 4.907±4.992 mm for the urethra, and 0.377±0.244, 14.708±9.489 mm, and 4.260±4.092 mm for the seminal vesicle. For the AI-R contours, the values were 0.757±0.072, 17.562±7.240 mm, and 3.050±1.339 mm for the prostate, 0.162±0.105, 19.956±11.962 mm, and 4.798±3.879 mm for the urethra, and 0.451±0.219, 16.069±9.245 mm, and 4.075±3.734 mm for the seminal vesicle. For the U-R contours, the values were 0.769±0.070, 15.109±6.177 mm, and 2.733±1.265 mm for the prostate, 0.144±0.091, 13.560±7.087 mm, and 3.406±1.985 mm for the urethra, and 0.471±0.216, 12.981±6.756 mm, and 3.264±2.308 mm for the seminal vesicle. Using the Wilcoxon signed-rank test with statistical significance defined as p<0.01, the AI-U Dice similarity for the prostate differed significantly from both the AI-R and U-R comparisons. Similarly, AI-U demonstrated significantly different HD values than both AI-R and U-R, whereas the AI-R and U-R HD comparison was statistically insignificant. All pairwise MDA comparisons for the prostate were statistically significant. For the urethra, the AI-U Dice similarity differed from the other two comparisons, while for HD, the AI-R and U-R comparisons differed significantly from each other. No significant differences were observed among the three comparisons for the seminal vesicle across any metric. The average times to produce contours for the AI, urologist, and radiologist were 96.5 seconds, 285.8 seconds, and 217.9 seconds, respectively (p<0.01 for each time comparison). Conclusion This study suggests that AI may be a useful tool for prostate segmentation workflows in streamlining MRI-US prostate cancer diagnostics by producing similar contouring results in less time. Additional investigation should be conducted regarding the differences between the pathological outcomes of AI and non-AI contours, and the accuracy, cost analysis, and efficiency of AI technology should be elucidated.
Stroke rehabilitation faces challenges such as limited accessibility and engagement, prompting interest in virtual reality-based interventions. While virtual reality shows promise, its effectiveness in improving quality of life (QoL) and upper extremity (UE) functions in stroke survivors remains unclear. To evaluate (1) the effectiveness of virtual reality-based interventions in QoL and UE functions among stroke survivors and (2) the feasibility and safety indicators across trials. A comprehensive search was conducted across six electronic databases: PubMed, EMBASE, Cochrane, Scopus, PsycINFO, and CINAHL to identify relevant articles published between Jan 2014 and December 2024. The search was updated on 9 May 2026. The Cochrane Handbook for Systematic Reviews of Interventions and the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were employed to ensure methodological rigor and standardized reporting. Two independent reviewers assessed the eligibility of the articles, and the quality of the studies were appraised using the Revised Cochrane Risk of Bias Assessment Tool for Randomized Trials (RoB2). Feasibility indicators (participation, adherence, attrition) and adverse events were also extracted. Thirty-four randomized controlled trials involving 1834 participants were included. Two studies had a low risk of bias, 19 had some concerns, and 13 had a high risk. Meta-analyses revealed non-significant effects of virtual reality-based interventions on QoL (SMD = -0.09, 95% CI -0.30 to 0.11, p = .36; low certainty) and UE functions assessed by upper extremity Fugl-Meyer (SMD = 0.56, 95% CI -0.02 to 1.15, p = .06; very low certainty) and Action Research Arm Test (SMD = 0.15, 95% CI -0.22 to 0,52, p = .41; low certainty). Narrative synthesis yielded mixed findings. Feasibility outcomes were generally favorable: participation among eligible patients was typically >80%, attrition <20%, and adherence exceeded 80% in the few trials reporting it. Adverse events were mainly mild and comparable to those in control groups. Current evidence does not demonstrate the superiority of virtual reality-based interventions over conventional rehabilitation for improving QoL or UE functions in stroke survivors, despite generally favorable feasibility and safety profiles. The low-to-very low certainty of evidence highlights the need for stronger methodological rigor, theoretical frameworks, and long-term follow-up. Virtual reality may serve as a complementary option when access to traditional rehabilitation is limited. PROSPERO (CRD42024619427).
Objectives: This study evaluated the effects of 660 and 940nm lasers on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). Materials and Methods: SHED were cultured and divided into six groups for laser irradiation: 660nm laser at 25mW with energy densities of 1 and 2J/cm2 for 8 and 16 seconds, and 940nm laser at 100mW with the same energy densities and durations. Two additional groups served as controls with no laser exposure. Cell viability was evaluated by the methyl thiazolyl tetrazolium (MTT) assay, and cell proliferation was measured by cell counting with a Neubauer chamber and qualitatively evaluated by 4',6-diamidino-2-phenylindole (DAPI) staining at 24 and 72 hours post- irradiation. Data were analyzed using one-way ANOVA, followed by Tamhane's and Tukey's tests. Results: Irradiation with 940nm laser enhanced cell proliferation compared to both 660nm laser and control groups at 24 and 72 hours (P<0.05), as confirmed by DAPI staining. At 24 hours, all laser-treated groups showed increased cell viability relative to the control, although the increase in the 660nm/2J/cm² group was not statistically significant (P>0.05). At 72 hours, only 940nm laser group showed significantly higher cell viability and proliferation than the control group (P<0.05). Conclusion: These findings suggest low-level laser therapy, particularly at the wavelength of 940nm enhances the viability and proliferation of stem cell, supporting its potential as a promising tool in regenerative medicine and tissue engineering.
Inflammatory bowel disease (IBD) is a chronic and progressive disorder of the digestive tract, including Crohn's disease and ulcerative colitis. Over the last decade, IBD has been extensively studied due to its association with dysregulated intestinal microbiota, increased mucosal permeability, and immune imbalance. Despite available pharmacological treatments, there is no definitive cure. Mucosal healing (MH) has emerged as a crucial therapeutic target, as it aims to restore the integrity of inflamed mucosae and is associated with improved clinical outcomes. However, most clinical studies rely primarily on clinical indices rather than direct endoscopic or histologic assessment of MH, and the optimal vitamin D (VD) dosing required to achieve true MH remains unclear. This review aims to examine the diverse functions of VD in modulating pathways relevant to MH within the context of IBD, highlighting its role in immune modulation, intestinal barrier integrity, and gut microbiota composition, and to discuss its potential therapeutic value in IBD management. Relevant experimental, translational, and human clinical studies focusing on VD status, supplementation, and mechanisms related to MH, immune regulation, and intestinal barrier function in IBD were considered. MH involves epithelial restitution, cell proliferation, and differentiation, and its achievement has been associated with reduced symptoms, lower relapse rates, and decreased need for surgical interventions. Beyond its traditional role in bone health, VD plays a critical role in preserving intestinal barrier integrity and regulating immune homeostasis. While biological evidence indicates that VD modulates immune responses, enhances tight junction integrity, and influences gut microbiota composition, clinical data primarily support its role in improving clinical and biochemical disease activity rather than definitively inducing mucosal repair. This review highlights VD as a relevant immunomodulatory and barrier-protective factor with potential complementary value in the clinical management of patients with IBD. Vitamin D and intestinal healing in inflammatory bowel disease Inflammatory bowel disease (IBD), which includes Crohn’s disease and ulcerative colitis, is a chronic condition that causes inflammation in the digestive tract. People living with IBD often experience abdominal pain, diarrhea, fatigue, and reduced quality of life. Although several treatments exist, there is currently no cure. One important treatment goal is “mucosal healing,” which means repairing the lining of the intestine and reducing inflammation to prevent relapses and complications. Vitamin D is best known for its role in bone health, but growing evidence shows that it also plays an important role in immune function and gut health. Many patients with IBD have low vitamin D levels, and this deficiency has been linked to more severe disease, increased hospitalizations, and higher relapse rates. This review summarizes current research on how vitamin D may help promote mucosal healing in IBD. Studies suggest that vitamin D helps strengthen the intestinal barrier, reduce harmful inflammation, and support a balanced gut microbiota. It also regulates immune responses and promotes the production of protective molecules that help repair intestinal tissue. Clinical studies indicate that vitamin D supplementation may reduce disease activity and improve quality of life in patients with IBD. However, optimal dosing remains unclear, as vitamin D levels can vary depending on factors such as diet, sun exposure, disease severity, and individual metabolism. Overall, maintaining adequate vitamin D levels appears to be a safe and promising complementary strategy for improving outcomes in patients with IBD. Future research is needed to determine the best supplementation strategies and to better understand how vitamin D contributes to long-term intestinal healing.
For decades, infections with the parasite Schistosoma mansoni have been treated with praziquantel, the only available drug, which highlights the urgent need of treatment alternatives. Natural compounds including rocaglates from plants of the genus Aglaia and pateamines isolated from the marine sponge Mycale hentscheli have exhibited promising anti-viral, anti-cancer, and anti-pathogenic properties. These compounds target the eukaryotic translation initiation factor 4A (eIF4A), which is a crucial factor for unwinding RNA structures in the 5'-UTR of selected mRNAs, and prevent the unwinding activity of eIF4A, which interferes with protein synthesis. Preliminary data indicated anthelminthic activity of the rocaglate silvestrol against S. mansoni in vitro. In this study, we confirmed the interaction of rocaglates and pateamines with recombinant S. mansoni eIF4A and analyzed the activity of these compounds against adult S. mansoni in vitro. Our findings showed reduced parasite vitality and stem-cell proliferation as well as impaired embryogenesis after rocaglate or pateamine treatment in the nanomolar range. Furthermore, we observed that rocaglate-treated S. mansoni recovered during washout experiments, while pateamine treatment displayed irreversible effects finally killing most of the worms. Although some properties of the used compounds impede their suitability as anti-schistosomal candidates, eIF4A remains a promising drug target in this parasite.
Critical-size bone defects remain a clinical challenge due to limited intrinsic bone regeneration. This study developed a composite scaffold composed of osteoinductive biphasic calcium phosphate (BCP) ceramic particles and gelatin methacryloyl (GelMA) hydrogel, loaded with calvarial osteoblasts (OBs) pretreated with the glycogen synthase kinase-3β (GSK-3β) inhibitor Tideglusib on surface of BCP particles, and evaluated its bone regenerative efficacy. BCP particles were characterized by X-ray diffraction and scanning electron microscopy. GelMA was synthesized and characterized by FTIR and 1H-NMR spectroscopy. Mechanical properties were assessed by compression testing. OBs were treated with Tideglusib to determine optimal concentration via CCK-8, scratch assays, qPCR, ALP and Alizarin Red S staining. Tideglusib-pretreated OBs seeded on BCP particles were evaluated for viability, proliferation, and osteogenic gene expression (COL-I, ALP). In vivo, 3.5 mm rat femoral defects were used to assess the bone regeneration potential of T-OB/BCP particle/GelMA (GPCT) composites in comparison with other groups by micro-CT and histology at 4 and 8 weeks. BCP (200-300 μm, HA/β-TCP ~30:70) incorporation enhanced GelMA compressive modulus ~4-fold. In vitro, 1 μM Tideglusib optimally activated Wnt/β-catenin signaling and promoted OB proliferation, migration, and osteogenic differentiation, with sustained effects after drug removal. OBs on BCP particles showed time-dependent upregulation of osteogenic genes, and Tideglusib pretreatment further elevated this expression. In vivo, micro-CT revealed significantly higher BV/TV, Tb.N, Tb.Th, and BMD in the GPCT group versus others. Histology showed more mature bone with numerous osteocytes, partial degradation of BCP particles, and new bone area fraction >70% in GPCT. The composite BCP particle/GelMA scaffold with Tideglusib-pretreated OBs provides mechanical reinforcement and sustained osteogenic stimulation, effectively accelerating critical-size bone defect regeneration. This ex vivo pharmacological priming strategy represents a promising translational approach for bone tissue engineering. Why was the study done? Repairing large bone defects remains a major clinic challenge. Current treatments, such as taking bone from another body part, cause additional injury and are limited in supply. A leading direction involves designing combined three elements: a supportive scaffold, bone-forming cells, and molecular signals that instruct cells to build new tissue. This study explored whether a scaffold that physically supports new bone growth—while delivering bone-forming cells temporarily “primed” with a bone-stimulating drug—could effectively repair large bone defects. What did the researchers do and find? We created a composite made of a gel reinforced with ceramic particles that naturally encourage bone growth. Bone-forming cells were briefly treated outside the body with a drug called Tideglusib, which activates a key bone-building pathway. The drug was then removed, leaving cells in a primed state without long-term drug exposure. This composite was implanted into a critical-sized leg bone defect in rats—a wound too large to heal on its own. After eight weeks, over 70% of the defect was filled with mature bone in treated animals, while untreated defects remained largely hollow. What do these results mean? This approach—combining a reinforced scaffold, bone-friendly particles, and temporarily boosted cells—can robustly regenerate bone in severe injuries. By briefly priming cells before implantation, we capture the benefits of a powerful bone-building drug while avoiding risks of long-term exposure. This strategy provides a foundation for developing next-generation bone graft for patients with severe bone loss.
Oral lichen planus (OLP) is a chronic autoimmune disease that negatively affects patients' quality of life through symptoms such as pain, irritation, bleeding, and ulceration. Despite numerous therapeutic approaches, there is no definitive cure, and conventional corticosteroid therapy is often associated with prolonged use and adverse effects. Photobiomodulation therapy (PBMT) has emerged as a promising non-invasive alternative with no reported side effects. This systematic review aimed to evaluate the efficacy of PBMT in the treatment of OLP. Electronic searches were conducted in PubMed, LILACS, Livivo, Scopus, Embase, Web of Science, and EBSCO up to March 1, 2025. Randomized and non-randomized clinical trials involving human participants were included, comparing PBMT with conventional pharmacological therapy. Risk of bias was assessed using the Cochrane tools, and the certainty of the evidence was evaluated using the GRADE approach. A total of 4,118 studies were identified, and 15 met the inclusion criteria after screening. Meta-analysis showed that PBMT slightly reduced pain levels compared with conventional therapy but did not significantly affect disease severity or anxiety scores. However, PBMT was associated with higher rates of complete mucosal healing and lower recurrence rates in patients with erosive OLP. PBMT appears to be a safe and effective therapeutic alternative for the management of OLP, particularly by reducing pain, promoting mucosal healing, and decreasing lesion recurrence. These findings support its potential role in clinical practice, although further high-quality randomized clinical trials are warranted to strengthen the current evidence.
To develop remineralizing dental composites incorporating prepolymerized TMPTMA-HEMA particles charged with CaF₂ (THCaF₂). Blends of TMPTMA-HEMA (50/50 wt%), charged with CaF₂ (5, 10, and 15 wt%), were heat-polymerized and crushed to a micrometric scale (THCaF₂). Four dental composites were produced using the same organic matrix (30 wt%) of Bis-GMA/TEGDMA (70/30 wt%). The control composite (C) contained 70 wt% barium borosilicate (BaBSi) fillers, whereas, in CaF5, CaF10, and CaF15, BaBSi fillers were partially replaced by 20 wt% of THCaF₂ particles. The remineralizing potential was evaluated through F⁻ release and the remineralization of caries-like enamel lesions induced by S. mutans biofilm using micro-CT. Physicomechanical properties included degree of conversion (DC%), flexural strength (FS), flexural modulus (FM), water sorption (Wsp), and water solubility (Wsl). Data were analyzed using one-way ANOVA and Tukey's HSD test (α = 0.05). All composites containing THCaF₂ released F⁻ ions, whereas only CaF10 and CaF15 induced enamel remineralization. The DC% did not differ among the dental composites (p > 0.05). C showed the highest FS and FM (p < 0.05), whereas the FS and FM of the other composites did not differ from one another (p > 0.05). The lowest Wsp was observed for C (p < 0.05), followed by CaF5, CaF10, and CaF15, which did not differ statistically from one another (p > 0.05). No statistical difference in Wsl was found among the dental composites (p > 0.05). The incorporation of prepolymerized particles charged with CaF₂ (THCaF₂) conferred remineralizing potential to experimental dental composites and represents a promising strategy for developing dental composites capable of preventing enamel demineralization caused by the caries process.
Ozonated water is increasingly being explored as a gentler alternative to chlorhexidine for chemical plaque control because of its antimicrobial and anti-inflammatory properties and the absence of common side effects such as staining or altered taste. However, the evidence on its effectiveness-especially in healthy individuals-remains unclear. This systematic review and meta-analysis aimed to determine whether ozonated water is more effective than placebo or chlorhexidine mouthwash in reducing dental plaque. An extensive literature search was carried out across PubMed, Cochrane Library, Scopus, Web of Science, LILACS, ScienceDirect, EBSCOhost, and Google Scholar up to August 5, 2025. Randomized controlled trials involving healthy participants and comparing ozonated water with placebo or chlorhexidine were included. In total, five studies were selected for qualitative synthesis, of which four were included in the meta-analysis. Risk of bias was assessed using the RoB 2 tool, and the certainty of evidence was evaluated using the GRADE approach. A random-effects model was applied to calculate standardized mean differences (SMDs) with 95% confidence intervals. The meta-analysis suggested that ozonated water may provide slightly better short-term plaque reduction than placebo or chlorhexidine (SMD = -1.35; 95% CI: -2.77 to 0.06). However, as the confidence interval crossed the line of no effect, this difference was not statistically significant. Risk of bias was high in one study, unclear in three studies, and low in one study. The overall certainty for plaque reduction was rated as "low". While ozonated water appears to be a promising short-term option for plaque control-especially for individuals unable to tolerate chlorhexidine-the current evidence is not strong enough to confirm that it works better than standard rinses or even plain water. Well-designed, longer-term clinical trials are needed before ozonated water can be recommended for routine daily use.
Ovarian cancer remains one of the most lethal gynecological malignancies due to late diagnosis, limited treatment options, and high recurrence rates. Recent evidence highlights the ubiquitin-proteasome system as a key contributor to cancer progression, with ubiquitin-conjugating enzyme E2 J1 (UBE2J1) emerging as a potential oncogenic target. In this study, a comprehensive in silico drug discovery approach was applied to identify natural phytochemicals capable of inhibiting UBE2J1. Sequence and structural modeling confirmed the reliability of the predicted protein conformation, while protein-protein interaction analysis underscored its central role in ubiquitination pathways. Five plant-derived compounds were screened through molecular docking, among which Withaferin A exhibited the strongest binding affinity (- 8.4 kcal/mol), forming stable hydrogen bonds and hydrophobic interactions with key active site residues. Molecular dynamics simulations further demonstrated the stability of the protein-ligand complex, with favorable RMSD, RMSF, and radius of gyration profiles. ADMET and drug-likeness evaluations revealed that Withaferin A complies with Lipinski's rule of five, possesses good intestinal absorption, and shows non-carcinogenic and non-mutagenic properties. Binding free energy calculations using MM/GBSA supported its strong affinity toward UBE2J1. These findings highlight Withaferin A as a promising natural inhibitor of UBE2J1 and provide a foundation for future experimental validation aimed at developing targeted therapies against ovarian cancer.
Oncogene SETDB1, an H3K9 methyltransferase, drives tumorigenesis in various cancers. Using endometrial cancer (EC) as a model, we discovered SETDB1's dual mechanisms in driving EC tumorigenesis and mediating immune evasion. SETDB1 knockout (SETDB1-/-) tumor-bearing mice exhibited prolonged survival up to 100 days. Transcriptomic profiling of SETDB1-/- EC cells revealed decreased oncogene expression and increased tumor suppressor gene expression, which indicates that SETDB1 intrinsically promotes EC proliferation by regulating these downstream genes. SETDB1 repressed repeat elements and the interferon pathway, mediating immune evasion extrinsically by inhibiting anti-tumor macrophage infiltration. ChIP-seq analysis showed SETDB1 binding at pericentromeric regions on many chromosomes and numerous ZNFs. Loss of SETDB1 resulted in abnormal cell division. SETDB1-/- tumors displayed reduced proliferation markers (Ki67, pHH3) and increased macrophage infiltration. Mechanistically, SETDB1 promotes CD47 (a don't-eat-me signal) and represses CCL5 and CXCL9 (macrophage and T-cell recruiting chemokines), contributing to immune evasion. M1-like macrophages killed more SETDB1-/- cells in co-culture. Additionally, SETDB1 knockout in mouse EC cells reduced tumor growth in C57BL/6 mice, with increased macrophage and CD4 + T-cell infiltration. Our results indicate that elevated SETDB1 and its targets can predict higher tumor grade and worse survival, suggesting that targeting SETDB1 could be a promising therapeutic strategy for EC.
Introduction Diabetes is among the many known chronic diseases resulting from the inability of the body to produce insulin. The use of traditional drugs such as sulfonylurea, biguanide, and thiazolidinediones has been associated with some side effects, hence making researchers turn their attention toward herbal medicine as a treatment or prevention option for diabetes. Hence, the present study was designed to evaluate and compare the metabolic effects of allantoin (5 mg/kg and 10 mg/kg), metformin (500 mg/kg), and their combination on blood glucose and body weight in low-dose streptozotocin (STZ)-induced diabetic Wistar rats. Methods An in vivo study employed a randomized, controlled, parallel-group, preclinical experimental design consisting of a total of 36 young adult (8-10 weeks old) male Wistar rats (Rattus norvegicus) weighing 200-260 g, which were randomly assigned to six experimental groups, namely, G1 to G6. G1 and G2 served as non-diabetic and diabetic controls administered intraperitoneally (ip) with 2 ml of normal saline (NS) and STZ 45 mg/kg + 2 ml NS, respectively. G3, G4, G5, and G6 groups served as treatment groups and received STZ 45 mg/kg, concurrently administered intraperitoneally with allantoin (ALN) 5 mg/kg, ALN 10 mg/kg, metformin (MET) 500 mg/kg (oral), and ALN 5 mg/kg (ip) + MET 500 mg/kg (oral), respectively. Fasting blood sugar (FBS) estimation was done for nine days daily after STZ injection, after diagnosis of diabetes mellitus on the 9th/10th day, and on the 28th day of treatment. Body weight of experimental animals was recorded once weekly on days 0, 7, 14, 21, and 28. Results FBS levels differed significantly among groups at baseline and day 28 (p < 0.0001). Diabetic controls maintained persistently elevated glucose levels, whereas ALN treatment at 5 mg/kg (301.17 ± 11.18 vs. 218.33 ± 10.11; -82.84; p < 0.001) and ALN 10 mg/kg (302.83 ± 5.08 vs. 171.17 ± 7.88; -131.66; p < 0.001) significantly reduced FBS in a dose-dependent manner. Metformin monotherapy markedly improved glycemic control (297.00 ± 17.73 vs. 126.00 ± 10.10; -171.00; p < 0.001), while combination therapy with ALN and metformin produced the greatest reduction in glucose levels (302.50 ± 11.41 vs. 120.00 ± 9.57; -182.50; p < 0.001), suggesting additive antihyperglycemic effects. ALN 10 mg/kg significantly increased body weight (p = 0.002). Conclusion Allantoin, particularly along with metformin, offers promising therapeutic potential for the management of diabetes and associated metabolic alterations.