Acute kidney injury (AKI) is a severe clinical syndrome, with ischemia/reperfusion (I/R) being one of its most common causes. Although D‑pinitol (DP), an inositol‑like bioactive molecule, is known to confer renal protection, its efficacy against I/R‑induced AKI remains unknown. A mouse kidney I/R model was employed to evaluate the renoprotective effect of DP. Relevant targets associated with DP and AKI were retrieved from publicly available databases. Subsequently, network pharmacology analysis was conducted to identify the potential targets and signaling pathways. Molecular docking was then performed to predict the binding affinity of DP to core targets identified. Furthermore, in vivo and in vitro experiments were performed to validate these findings. Systemic toxicity was assessed by serological and histopathological examinations. The results show that DP significantly attenuated I/R-induced kidney dysfunction and apoptosis. Network pharmacology analysis identified 108 overlapping targets, with AKT1, HSP90AA1, SRC, CASP3, and MMP9 identified as core targets. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed PI3K/AKT signaling pathway as the primary mechanism. Consistent with these predictions, DP enhanced PI3K and AKT phosphorylation in kidney tissue. Molecular docking indicated that DP exhibited the strongest binding affinity to SRC, suggesting it as a potential target. In a hypoxia/reoxygenation (H/R)-induced human renal proximal tubular epithelial (HK-2) cell model, DP significantly increased the phosphorylation of SRC, PI3K, and AKT, and these effects were abrogated by the SRC specific inhibitor PP2. Collectively, DP alleviated I/R‑induced injury and apoptosis, potentially through activation of the SRC/PI3K/AKT signaling pathway. Acute kidney injury (AKI) is a serious and potentially life-threatening condition that currently lacks specific and effective treatments. This study investigated whether D-pinitol (DP)—a natural compound found in soybeans and other legumes—can protect the kidney from damage caused by interrupted blood flow (a condition known as ischemia/reperfusion injury, I/R injury). Using computer-based prediction tools (network pharmacology and molecular docking), we first identified the potential molecular targets and protective pathways of DP. We then confirmed these predictions through laboratory and animal experiments. Our results showed that DP significantly improved renal function and reduced tissue damage after I/R injury. Mechanistically, DP activated the SRC/PI3K/AKT cell survival pathway, which effectively reduced the death of renal tubular cells. As a safe and naturally occurring compound, DP represents a promising new preventive strategy for patients at high risk of developing AKI.
This study aimed to detect the prevalence, antimicrobial susceptibility, and molecular characterization of some virulence determinants in Salmonella recovered from 100 diseased turkeys in Dakahlia Governorate, Egypt. Twelve Salmonella isolates were recovered from the diseased turkeys with an overall prevalence of 12%. The recovered isolates were serotyped into 4 serovars: (S. enterica serovar Enteritidis, S. enterica serovar Typhimurium, S. enterica serovar Infantis and S. enterica serovar Heidelberg). Additionally, a 21-d trial investigated the effects of dietary Mannan-oligosaccharides (MOS) and β-glucans as feed additives on growth performance parameters, intestinal morphology, and blood biochemical, immunological parameters, and fecal shedding of S. enterica serovar Enteritidis-challenged turkey. A total of 300 One-day-old turkey poults were allocated into five treatment groups (100 poults for each) in a completely randomized design: group 1: a negative control group (non-challenged, non-supplemented), group 2: a positive control group (S. enterica serovar Enteritidis-challenged at 7th day, non-supplemented), group 3 and 4: two challenged groups supplemented with a mixture of MOS and β-glucan at 1 & 2 g/kg diet, respectively, and T5: a challenged group and treated with Florfenicol antibiotic on the 8th day of the experiment at a dose of 1 ml /liter drinking water for five consecutive days. The in vivo experimental results showed that birds in the positive control group showed reduced growth performance compared with the negative control group. Supplementation with MOS and β-glucan, particularly at 2 g/kg ration, significantly improved body weight gain, and feed conversion ratio, in addition to reduced clinical signs and mortality compared with the positive control group. Overall, MOS and β-glucan at 2 g/kg restored growth performance close to negative control and showed better feed efficiency than the antibiotic-treated group. MOS and β-glucan supplementation, particularly at 2 g/kg, significantly increased globulin, thyroid hormones, and immunoglobulins (IgA, IgG, and IgM), and IL1β, and interferon-gamma (IFN-γ) compared with the control groups. Histopathological examination revealed normal liver and spleen structures in the negative control group, while the positive control group showed marked degenerative and inflammatory changes in the liver and lymphoid depletion in the spleen. Supplementation with MOS and β-glucan, particularly at 2 g/kg, markedly improved hepatic and splenic histological structures with mild inflammatory changes, approaching the normal architecture. The antibiotic-treated group also exhibited nearly normal histological features. Bacteriological examinations revealed lower Salmonella shedding in the MOS and β-glucans 2 g/kg group in comparison with the positive control and the MOS and β-glucans 1 g/kg groups. In conclusion, dietary addition of MOS and β-glucans 2 g/kg in turkey diets improved growth performance, immune status, and hepatic and splenic histological structures in addition to lowering Salmonella fecal shedding.
Diabetic hepatic injury is a common complication of diabetes, yet it is often neglected due to its subtle early symptoms. This study explored the effect and mechanism of Anaphalis virgata extract (AVE) in treating diabetic hepatic injury. Serum-exposed components of AVE were identified by LC-MS and used for network pharmacology to predict therapeutic targets and pathways. AVE was evaluated in diabetic rats treated intragastrically at 50, 100, and 200 mg/kg for 10 weeks. Hepatic biochemical parameters were measured, and mechanisms were verified by molecular dynamics (MD) simulation, ELISA, and Western blot. Thirty-one serum-exposed compounds (8 prototypes and 23 metabolites) were identified, primarily flavonoids, phenolic acids, terpenoids, and phenylpropanoids. Network pharmacology revealed pathways including lipid and atherosclerosis, IL-17, and AGE-RAGE signaling. Apigenin, kaempferol, and caffeic acid were identified as core compounds, and IL-6, TNF, and IL-1β as core targets. In vivo, AVE significantly decreased hepatic IL-6, TNF, IL-1β, TC, TG, FFA, AST, and ALT levels in diabetic rats. Western blot showed that AVE up-regulated AKT and AMPK while down-regulating NF-κB and GSK-3β pathways. AVE exerts a therapeutic effect on diabetic hepatic injury through anti-inflammatory mechanisms, providing a foundation for its further development and clinical application.
Cancer development arises from dynamic interactions between inherited susceptibility and modifiable environmental exposures, among which diet plays a central role. Carotenoids, lipophilic plant-derived pigments including lycopene, α-carotene, and β-carotene, and retinoids, the vitamin A derivatives that regulate gene transcription via retinoic acid receptors (RARs) and retinoid X receptors (RXRs), have been extensively investigated for their chemopreventive and therapeutic potential. This review aims to provide an integrated, mechanism-based synthesis of the roles of lycopene, α- and β-carotene, and retinoids in cancer chemoprevention and to clarify the conditions under which they are most likely to be effective. Beyond summarizing established antioxidant and nuclear-receptor mechanisms, we highlight as a novel emphasis the epigenetic actions of these compounds, including effects on DNA methylation, histone modification, and microRNA regulation, and we integrate these with the well-recognized divergence between dietary and high-dose supplement outcomes. Experimental evidence demonstrates that carotenoids modulate oxidative stress, inflammation, proliferation, apoptosis, angiogenesis, and metastasis through pathways such as Nrf2/ARE, NF-κB, STAT3, Akt/mTOR, MAPK, and Wnt/β-catenin. Lycopene, in particular, exhibits strong antioxidant capacity and multi-target signaling effects, while provitamin A carotenoids additionally influence retinoid-mediated transcriptional programs. Retinoids exert broader differentiation-inducing and antiproliferative effects through direct nuclear receptor signaling and represent one of the few successful differentiation therapies in oncology, most notably in acute promyelocytic leukemia. Epidemiologic studies generally associate higher dietary carotenoid intake with reduced risk of several malignancies, including prostate, breast, lung, colorectal, and gastric cancers. However, randomized trials of isolated high-dose supplementation, particularly β-carotene in smokers, have demonstrated null or harmful effects, highlighting a critical divergence between whole-food dietary patterns and pharmacologic supplementation. In conclusion, carotenoids and retinoids possess biologically plausible anticancer properties, yet their clinical utility remains context dependent. Future research should prioritize biomarker-guided, precision-based strategies, standardized formulations, and whole-food dietary approaches to clarify their role in cancer prevention and treatment.
Elizabethkingia species have emerged as important nosocomial pathogens associated with multidrug resistance and persistent infections. This study aimed to characterize clinical Elizabethkingia isolates from Northern Thailand regarding antimicrobial susceptibility, virulence-associated phenotypes, and biofilm formation, and to evaluate the antimicrobial and anti-biofilm activity of dialdehyde cellulose (DAC) film. A total of 49 clinical isolates were identified by MALDI-TOF mass spectrometry, with species identification confirmed by 16S rRNA gene sequencing. Antimicrobial susceptibility was determined against 12 agents. Virulence traits (protease, lipase, lecithinase, and hemolysin production) and biofilm formation were assessed using standard phenotypic assays. DAC films were evaluated against selected resistant isolates. Elizabethkingia anophelis predominated, and most isolates exhibited multidrug or extensive drug resistance, with high resistance to carbapenems and cephalosporins. Piperacillin-tazobactam, levofloxacin, and trimethoprim-sulfamethoxazole showed the greatest activity. All isolates demonstrated protease production and time-dependent hemolysis, while lipase and lecithinase activities were absent. Biofilm formation varied among isolates, while DAC films inhibited bacterial growth and prevented detectable biofilm formation in the tested isolates. No significant difference was observed between DAC and DAC supplemented with meropenem in inhibition zone diameters (p = 0.555). Clinical Elizabethkingia isolates demonstrated extensive antimicrobial resistance with conserved virulence traits and heterogeneous biofilm formation. DAC films demonstrated antimicrobial activity and prevented detectable biofilm formation under the experimental conditions. Further studies are warranted to evaluate their mechanism of action and potential applications.
The combination of bevacizumab and TAS-102 has emerged as a treatment option for metastatic colorectal cancer (CRC). Bevacizumab is an anti-VEGF-A antibody, and TAS-102 is a cytotoxic nucleotide analog. Recently, fruquintinib, an oral, potent and highly selective inhibitor of all three vascular endothelial growth factor receptors (VEGFRs), has been approved for the treatment of chemorefractory CRC. The purpose of this study was to investigate the combined antitumor activity of fruquintinib and TAS-102 in CRC and gastric cancer (GC) models and to reveal possible mechanisms to explain the combined effects. In vitro angiogenic and lymphangiogenic activities were studied by tube formation assays using HUVECs and HDLECs, respectively. Protein expression was measured by Western blot analysis. Intratumor drug concentrations were determined by liquid chromatography-tandem mass spectrometry analysis. Antitumor activity was studied in xenograft models. Fruquintinib inhibited tube formation in both HUVECs and HDLECs. Trifluridine, an anti-neoplastic component of TAS-102, potentiated the inhibition of tube formation by fruquintinib in HUVECs but not in HDLECs. Suppression of VEGFR downstream signaling, together with induction of DNA damage response, was observed in HUVECs treated with the fruquintinib plus trifluridine combination. In an HT-29 CRC xenograft model, the intratumor levels of trifluridine and its active metabolites were higher following the fruquintinib plus TAS-102 combination treatment than following TAS-102 monotherapy. Antitumor studies demonstrated that the combination of fruquintinib and TAS-102 suppressed tumor growth more potently than individual agents in HT-29 and COLO-205 CRC xenograft models. The combined effect was comparable to that of bevacizumab plus TAS-102 in CRC models and was also observed in SC-2-JCK and MKN45 GC xenograft models. The addition of fruquintinib did not increase body weight loss caused by TAS-102 alone. The combination of fruquintinib and TAS-102 improved antitumor activity in CRC and GC xenograft models, with in vitro evidence of enhanced anti-angiogenic activity of fruquintinib by trifluridine and ex vivo evidence of an increased amount of intratumor trifluridine with fruquintinib. Our data support testing the combination of fruquintinib and TAS-102 in clinical studies.
Hypothyroidism has been associated with cognitive disorders and high risk of neurodegenerative disorders like Alzheimer's disease (AD), which is caused by amyloid-β (Aβ) accumulation in brain. The aim of the study is to examine the therapeutic ability of triiodothyronine (T3) and curcumin in regulating two key extracellular matrix proteins (ECM), collagen IV and collagen VI in brain since both drugs have neuroprotective and anti-fibrotic effects. The study further explored the possible role of signaling molecules such as transforming growth factor β1 (TGFβ1) and specificity protein 1 (Sp1) in regulating the effects of T3 and curcumin on Aβ accumulation and ECM protein expression. Experiment was carried on Balb/c mouse model by inducing hypothyroidism with 0.05% PTU followed by treatment of T3, curcumin and a combination of both. The study revealed a significant increase in the expression of collagen IV and VI, along with elevated levels of TGFβ1 and Sp1 signalling proteins, in hypothyroid conditions in response to increased Aβ expression in both the cortex and hippocampus. Western blot and RT-PCR analysis showed that T3 and curcumin treatment downregulate the levels of ECM proteins, the APP gene, Aβ protein, and the signaling factors. The study found that T3 and curcumin treatment downregulates the collagen IV and VI expression by negatively regulating TGFβ1 and Sp1 when expression of APP and Aβ are low. T3 and coadministration of T3 with curcumin more effectively regulate thyroid function and ECM protein expression in both the cortex and hippocampus. Whereas curcumin alone exhibits limited impacts under the experimental conditions in our study, this may due to bioavailability, dosage or treatment duration which need to further investigation. Our results indicate a novel therapeutic approach for understanding Aβ pathology associated with hypothyroidism, emphasizing the potential association between thyroid hormone and nature-based compounds, such as curcumin, in modulating ECM dynamics and neuroprotection.
The gut microbiota comprises a diverse and dynamic community of microorganisms that collectively enhance host metabolism, physiology, and overall functionality. In this context, the swine archaeome remains largely underexplored despite growing evidence that archaea may greatly influence host health. Advances in high-throughput approaches provide new opportunities to reveal the dynamics and composition of archaea. Herein, we uncover the taxonomic and functional landscape of the piglet archaeome during the weaning transition under multiple experimental conditions, integrating shotgun metagenomic and metatranscriptomic analyses to elucidate its contribution to gut microbial ecology. The seven experimental conditions included four antibiotic treatments for post-weaning diarrhoea (trimethoprim/sulfamethoxazole, colistin, gentamicin, amoxicillin), an oral vaccine, acidifiers in drinking water, and a no-intervention group. A total of 280 faecal samples were collected longitudinally one day before weaning (ST1), three days (ST2), two weeks (ST3), and four weeks (ST4) after the start of the treatment. Treatment was initiated eleven days after arrival at the experimental farm following the onset of clinical signs. Shotgun metagenomics was used to assess archaeal taxonomic diversity and recover archaeal metagenome-assembled genomes (aMAGs), while metatranscriptomics was integrated to assess differentially expressed genes at ST1, ST2, and ST4. The results revealed archaea as the second most abundant microorganism, exhibiting a longitudinal increase in diversity over the experimental time. The most predominant genus was Methanobrevibacter, including Methanobrevibacter smithii. Eleven high-quality aMAGs were recovered, belonging to the Methanobacteriota and Thermoplasmatota phyla. Genome-inferred functional analyses revealed that the predominant metabolic processes included the biosynthesis of nucleic acids, amino acids, organic anions, and vitamins. Additional functional traits suggested potential roles in the degradation of sugars, amino acids, and antibiotics were also observed. Moreover, significant differences were detected on the archaeal metatranscriptome between the experimental groups treated with antibiotics and the rest of the groups, underscoring their response to changes in microbial interactions, substrate availability and, in some cases, direct effect of the antimicrobials on metabolic pathways. Altogether, this study highlights the biological significance of archaeal dynamics during initial life stages and demonstrates how combining metagenomics and metatranscriptomics uncovers their functional potential and the pathways actively expressed in the piglets' gut.
Background and Objectives: Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease in which persistent synovial inflammation and joint damage are influenced not only by immune dysregulation but also by environmental, genetic, and epigenetic factors. Vitamin D is a secosteroid hormone and has emerged as a key immunomodulatory hormone, with reported effects on innate and adaptive immune responses, with potential relevance to RA clinical activity and treatment. This narrative review synthesizes mechanistic and clinical evidence enlightening vitamin D's immunomodulatory role in RA pathogenesis and management. Methods: A comprehensive literature search was carried out on PubMed and MEDLINE databases using Medical Subject Headings (MeSH) terms: "Vitamin D", "Cholecalciferol", "Arthritis, Rheumatoid", "Seasons", "Epigenomics", "DNA Methylation", and "Therapy". The narrative review highlights evidence published mainly in the last 5 years on the link between vitamin D and RA, focusing on epigenetic interactions, circannual rhythms, and therapeutic implications. Results: Emerging data suggest that vitamin D-related epigenetic mechanisms (e.g., DNA methylation, histone acetylation, and microRNA regulation) and genetic polymorphisms have been associated with disease susceptibility and treatment outcomes. Latitude and seasonal fluctuations in serum 25-hydroxyvitamin D levels correlate with variations in RA disease activity, although results remain heterogeneous across studies. Overall, the available evidence supports an association between vitamin D deficiency and greater RA disease activity, while its adequate supplementation has been associated with improvements in inflammatory markers and selected clinical outcomes, especially when tailored to baseline status and individual risk factors, such as limited dietary intake and sunlight exposure. Conclusions: Current evidence emphasizes the need for further studies using standardized methods and larger, geographically diverse cohorts to define how best to leverage seasonal vitamin D variations in RA management, considering also the range of concomitant epigenetic modifiers that may influence the effects of vitamin D on the management of RA patients.
Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by persistent pain and a significant decline in quality of life. Tetrandrine (Tet), a bisbenzylisoquinoline alkaloid, exhibits significant antipyretic, analgesic, and anti-inflammatory effects. However, its low solubility in water limits its clinical applications. Objective: This study focuses on the preparation of tetrandrine nanocrystals (Tet-NCs) using ultrasound technology and evaluates their therapeutic effects and potential mechanisms in a rat model of adjuvant arthritis (AA). The AA model was induced using Freund's complete adjuvant (FCA). Tet-NCs were prepared via ultrasound technology and administered at varying doses. The efficacy of Tet-NCs was compared with that of Tet tablets and indomethacin tablets. The assessed outcomes include joint swelling, arthritis scores, pathological changes in the joints, levels of inflammatory factors (interleukin-6, interleukin-1β, tumor necrosis factor-α, prostaglandin E2, cyclooxygenase-2, vascular endothelial growth factor A, and matrix metalloproteinase-3) in joint cavities and serum, as well as the angiogenesis in synovial tissues and the expression of key proteins-Janus kinase 1 (JAK1) and signal transducer and activator of transcription 3 (STAT3). Experimental data indicate that the average particle size of the optimized Tet-NCs is 124.30 nm, with a polydispersity index (PDI) of 0.27. The therapeutic effects of low-dose Tet-NCs were comparable to those of Tet tablets, while medium- and high-dose Tet-NCs were significantly more effective than Tet tablets in reducing ankle joint swelling, lowering inflammatory factors levels, and protecting joint structure, and showed no statistically significant difference from indomethacin under the tested conditions. Furthermore, the expression levels of JAK1 and STAT3 in the AA model were significantly reduced by both medium- and high-dose Tet-NCs (all P < .001). This study demonstrates that Tet-NCs provide superior therapeutic effects in RA compared to conventional Tet tablets, and their mechanism of action may involve the regulation of inflammation through inhibition of the JAK1/STAT3 signaling pathway. Consequently, Tet-NCs show potential for further development as a novel anti-arthritic agent, although their clinical applicability remains to be validated in future studies. Clinical trial number: not applicable.
Background/Objectives: Extra virgin olive oil (EVOO), a key component of the Mediterranean diet, has attracted research interest because olive-derived phenolics demonstrate potential anticancer activity in experimental models. This review summarizes evidence concerning whole EVOO, phenolic-enriched EVOO, olive phenolic extracts, and the isolated compounds hydroxytyrosol, oleuropein, oleocanthal, and oleacein. Methods: A structured narrative search of PubMed, Web of Science, ScienceDirect, and Google Scholar was conducted for literature published between 2015 and 2025. Evidence was reviewed for breast, prostate, colorectal, pancreatic, bone, oral, liver, gastric, hematological, and brain cancers. Comparatively limited evidence concerning cervical, endometrial, ovarian, melanoma, non-melanoma skin, and thyroid cancers was summarized separately. Results: The molecular evidence was derived primarily from cell culture and animal studies using isolated phenolics and concentrated extracts. Preclinical studies indicate that EVOO phenolics may demonstrate anticancer activity through multiple mechanisms, including antioxidant activity, anti-inflammatory effects, cell cycle arrest, induction of apoptosis, inhibition of metastasis, anti-angiogenic activity, and modulation of key signaling pathways, such as PI3K/AKT/mTOR, MAPK/ERK, NF-κB, JAK/STAT, Wnt/β-catenin, p53, and epithelial-mesenchymal transition-related pathways. Most molecular and pathway-level evidence was obtained using isolated phenolic compounds in cell culture or animal models, whereas evidence directly examining whole EVOO consumption was largely observational and substantially more limited. Experimental studies also reported that oleocanthal induced lysosomal membrane permeabilization, whereas hydroxytyrosol and oleuropein promoted mitochondria-mediated apoptosis. Furthermore, preclinical combination studies suggested enhanced tumor-cell sensitivity to selected chemotherapeutic, targeted, and immunotherapeutic agents. However, these effects have not been established in patients. Human evidence remains limited mainly to observational dietary associations and small exploratory interventions, with no conclusive demonstration of cancer prevention or therapeutic efficacy. Conclusions: Isolated EVOO-derived phenolic compounds demonstrated promising anticancer mechanisms in preclinical models. However, these results should not be directly extrapolated to dietary EVOO because experimentally administered doses, bioavailability, metabolism, and food-matrix interactions differ substantially from human dietary exposure. Therefore, well-designed studies using chemically characterized EVOO, pharmacokinetic investigations, and controlled human trials are required before dietary or clinical recommendations can be made.
Healing a skin wound is a complex biological process that proceeds through three stages: inflammation, proliferation, and remodeling. Hyaluronic acid (HA) and vitamin C may help tissues heal in different but complementary ways. This study aimed to examine and compare the effects of HA and vitamin C, alone and in combination, on collagen-related histological and immunohistochemical repair indices at day 22 in a rat sutured incisional wound model. Twenty adult male albino rats were divided into four equal groups (n = 5/group) at random: the control, vitamin C-treated, HA-treated, and combined HA + vitamin C-treated groups. Each animal received a standardized sutured full-thickness cutaneous incision to model primary-intention wound healing. Treatments were administered daily for 21 days using different administration routes: topical HA, oral vitamin C, or their combined regimen; wound tissue was collected on day 22 to evaluate late histological and immunohistochemical repair outcomes after completion of the treatment period. Hematoxylin and eosin, Masson's trichrome, and collagen type I immunohistochemistry were used to evaluate collagen-related histological and immunohistochemical repair features. Collagen type I expression, collagen deposition, and fibroblast count were quantified by morphometry. The H&E-based fibroblast-like cell count showed an exploratory between-group difference in the primary ANOVA analysis (p = 0.039), with the HA + vitamin C group having the highest mean; however, pairwise significance was limited to the comparison with the water-treated group, and the non-parametric sensitivity analysis was more conservative. Collagen deposition was higher in the combined-treatment group than in the water, HA-alone, and vitamin C-alone groups, and the difference was significant (p < 0.001). Additionally, collagen type I expression varied significantly (p = 0.003), with the HA + vitamin C group having the highest mean area percentage and significant pairwise differences compared to the water and vitamin C groups. At day 22, the combined HA + vitamin C regimen showed the highest collagen deposition and collagen type I immunopositive area among the tested groups, whereas the fibroblast-like cell-count finding was less robust. These preliminary findings are limited to late histological and immunohistochemical outcomes in a small sutured-incision model and do not establish functional benefit or an HA-vitamin C interaction.
Periodontal disease (PD) is a chronic, biofilm-driven inflammatory condition in which connective tissue destruction and alveolar bone loss are driven largely by dysregulated host immunity. Prior experimental and clinical studies have linked IL-17 pathway activity to periodontal inflammation and treatment response, supporting its relevance as a therapeutic target in PD. Rather than targeting individual IL-17 isoforms, IL-17R blockade offers an alternative strategy because it serves as a shared downstream signaling node for multiple IL-17 family cytokines, potentially enabling broader modulation of inflammatory signaling. However, the therapeutic potential of IL-17R blockade in PD remains largely unexplored. Gingival and salivary immune profiles and inflammatory markers were assessed in periodontally healthy, diseased, and post-therapy subjects. For mechanistic studies, mice subjected to ligature-induced periodontitis (LIP) received local gingival injection of the IL-17 receptor inhibitor brodalumab at 5 or 20 mg/kg, or vehicle control every 48 h. Animals were euthanized on day 4 or day 8 post-treatment, and gingival inflammatory responses were quantified by qPCR and flow cytometry. Alveolar bone outcomes were assessed by micro-CT, including CEJ-ABC distance as a measure of bone loss, and bone volume and bone mineral density as measures of bone quality. Gingival and salivary CD4⁺IL-17⁺ T cells were increased in PD and were markedly reduced after therapy, supporting the association of IL-17-driven immune responses with disease activity and their resolution following treatment. This observation was validated in human gingival biopsies, where IL-17 expression was elevated in PD and markedly reduced after nonsurgical periodontal therapy, accompanied by lower inflammatory cytokine expression and a shift toward a healthy control profile. In LIP mice, local injection of brodalumab reduced gingival inflammatory transcripts, with IL-17A suppressed by day 4 and sustained through day 8, alongside reduced IL-17B, indicating broader dampening of IL-17 signaling. These changes were associated with reduced gingival neutrophil infiltration and M2-like macrophage skewing, indicating that brodalumab promotes the resolution of inflammation. Micro-CT demonstrated lower alveolar bone loss in a dose-dependent manner in brodalumab treated animals. Brodalumab also preserved bone quality, increasing bone volume and bone mineral density, most prominently at the higher dose, which approached no-ligature values by day 8. Local gingival administration of brodalumab attenuates IL-17-associated inflammatory responses and protects against alveolar bone loss in experimental periodontitis. Our findings support IL-17R blockade as a promising host-modulatory strategy that could serve as an adjunct to conventional periodontal care to limit periodontal tissue destruction.
Jianpi Bushen Qingchang Huashi Formula (JBQHF) is a traditional Chinese medicine compound of nine herbal ingredients clinically used to treat chronic diarrhea and dysentery syndromes, corresponding to ulcerative colitis (UC). Although clinical evidence supports its efficacy, the underlying mechanism remains unclear. This study sought to define the therapeutic benefit of JBQHF in a murine colitis model and to dissect the underlying mode of action. Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) was applied to profile the chemical constituents of JBQHF. Therapeutic efficacy was evaluated in a DSS-induced murine colitis model. Gut bacterial composition and metabolite output were analyzed by 16S rRNA gene sequencing and Gas chromatography-mass spectrometry (GC-MS), respectively. Network pharmacology, deep learning-based virtual screening, and molecular docking were employed to predict candidate active compounds and their potential targets. Immunological assays focused on Histone deacetylase 1 (HDAC1) expression, HDAC enzymatic activity, histone acetylation status and the T helper 17/regulatory T (Th17/Treg) differentiation axis. JBQHF attenuated colitis as reflected by lower disease activity indices (DAI) and reduced histopathologic injury. Pro-inflammatory mediators were decreased while intestinal tight junction protein levels were elevated. JBQHF was associated with increased gut microbiota-derived butyrate production, reduced colonic HDAC1 expression and HDAC enzymatic activity, elevated Acetyl-H3K9 and Acetyl-H3K27, and a rebalancing of the Th17/Treg ratio. The protective effects of JBQHF were largely abrogated by the HDAC activator ITSA-1. Deep learning-based virtual screening further identified flavonoids within JBQHF as candidate HDAC1 ligands with predicted target selectivity. JBQHF ameliorates experimental colitis via a dual inhibition of HDAC1, providing an integrated mechanism for its clinical application.
Background/Objectives: Anthracycline-induced cardiotoxicity remains a major challenge in cancer treatment, and researchers are showing interest in artificial intelligence (AI) to improve the prediction and detection of cancer therapy-related cardiac dysfunction (CTRCD). Current surveillance strategies rely mainly on left ventricular ejection fraction and, more recently, global longitudinal strain. Methods: The present study was designed to evaluate cardiac performance in a rat model of doxorubicin-induced cardiotoxicity and empagliflozin-mediated cardioprotection using a machine learning-based analytical framework. Eighteen adult male Sprague-Dawley rats were assigned to five experimental groups. We aimed to quantify ventricular wall dynamics and contractility using an advanced image-processing and object-detection model that has not been previously used to distinguish normal from impaired cardiac kinetics. During real-time recording, simultaneous electrocardiogram monitoring was performed, enabling direct correlation between deep learning-based ventricular wall motion metrics and cardiac electrical activity. The cardioprotective effects of empagliflozin were further validated by immunofluorescence staining (cTnI, vimentin, α-SMA, and Cx43) of rat cardiomyocytes and paraffin-embedded cardiac tissue, demonstrating attenuation of cellular injury and structural remodeling. Results: The integrated analysis of cardiac kinetic patterns derived via machine learning distinguishes not only extreme cardiotoxicity, but also tracks a graded pattern consistent with ECG-derived severity and treatment-related functional preservation. These findings indicate that the algorithm captures the gradient of empagliflozin's cardioprotective effect within this internally validated preclinical setting. Additionally, immunofluorescence results validated the benefits of SGLT2 inhibition on myocardial integrity. Conclusions: The novelty of the present work lies at the intersection of advanced cardiac kinetic analysis using AI, preclinical modeling, and SGLT2-mediated cardioprotection in cardio-oncology.
Alzheimer's disease (AD) is the most common neurodegenerative disorder and a leading cause of dementia worldwide, characterized by progressive cognitive decline, memory impairment, and neuronal loss. The pathological hallmarks of AD include extracellular accumulation of amyloid-β (Aβ) plaques, intracellular neurofibrillary tangles composed of hyperphosphorylated tau protein, chronic neuroinflammation, oxidative stress, mitochondrial dysfunction, and synaptic degeneration. Current symptomatic therapies provide modest clinical benefits, while recently approved amyloid-targeting monoclonal antibodies, such as lecanemab and donanemab, can slow decline in selected early-stage AD patients but do not cure the disease and are associated with safety, access, and cost concerns. This narrative review summarizes mechanistic evidence from in vitro and in vivo studies and distinguishes preclinical promise from validated clinical utility. Phytochemicals, including polyphenols, flavonoids, alkaloids, terpenoids, and carotenoids, demonstrate neuroprotective effects through antioxidant activity, anti-inflammatory modulation, inhibition of amyloid aggregation, regulation of tau phosphorylation, and support of mitochondria and synapses. Evidence from experimental models suggests that several phytochemicals may help slow AD pathology and improve cognitive function, but clinical translation remains limited due to poor bioavailability, inadequate blood-brain barrier (BBB) penetration, and a lack of large-scale clinical trials. This review highlights critical research gaps and emerging strategies to facilitate phytochemical-based preventive and therapeutic approaches in AD.
The elevated mortality associated with ovarian cancer arises from delayed detection, recurrent disease, and the rapid emergence of chemoresistance. This study assesses the anticancer efficacy of two synthetic curcumin analogues, B-143 and B-155, in comparison to natural curcumin, employing SKOV3 ovarian cancer cells as the experimental model. The aim of this study was to determine whether structural alterations to the analogues enhanced their functional performance. Through assays assessing cytotoxicity, cell-cycle distribution, apoptosis, and migration, the findings revealed that B-155 exhibited significantly greater cytotoxicity compared to curcumin and B-143, which was associated with G2/M cell-cycle arrest and increased apoptosis. Furthermore, both B-155 and curcumin effectively suppressed SKOV3 cell migration, whereas B-143 displayed minimal effects. Network pharmacology analyses predicted that B-143 and B-155 interact with overlapping yet distinct angiogenesis- and metastasis-associated signaling networks, suggesting potential associations with multiple signaling pathways relevant to ovarian cancer. Moreover, RT-qPCR reveals curcumin consistently downregulated the expression of genes linked to epithelial-mesenchymal transition (EMT) and cellular stress, whereas B-143 and B-155 displayed only partial adaptive responses in several angiogenesis and metastasis transcription markers. To further explore the molecular basis of B-155 activity, molecular docking and molecular dynamics simulations were performed using MAPK14 (p38α MAPK) as a candidate target. Computational analyses demonstrated favorable binding of B-155 within the MAPK14 ATP-binding pocket and stable protein-ligand complex formation throughout a 100 ns simulation. B-155 demonstrated more significant anticancer activity than B-143, suggesting superior structural change-enhanced effectiveness. This research highlights the influence of structural modifications on curcumin's biological activity, supports B-155 as a promising curcumin analogue for further investigation with anti-proliferative and migration suppressive properties, and emphasizes the need to incorporate both phenotypic and network-level assessments together with structural interaction analyses in curcumin-based drug development for ovarian cancer.
The selective inhibition of Bruton's tyrosine kinase (BTK) has emerged as a promising therapeutic strategy for autoimmune diseases by targeting key signaling pathways in both adaptive and innate immune cells. Initially developed for B cell malignancies, BTK inhibitors are now being evaluated across a range of immune-mediated disorders, including multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus. Beyond B cell receptor signaling, BTK inhibition modulates Fc receptor- and toll-like receptor-dependent activation of myeloid cells, thereby extending its therapeutic relevance. BTK inhibitors comprise a pharmacologically diverse class, including covalent (irreversible) and non-covalent (reversible) inhibitors, which differ in binding mode, selectivity, and pharmacokinetic properties. These differences are increasingly recognized as critical determinants of clinical performance. While early studies demonstrated robust target engagement and anti-inflammatory activity, clinical outcomes have been inconsistent across diseases. In multiple sclerosis, BTK inhibitors have shown promising effects on inflammatory activity and emerging signals on disability progression. In contrast, in rheumatoid arthritis and systemic lupus erythematosus, several compounds failed to meet primary clinical endpoints despite clear pharmacodynamic effects. These divergent outcomes highlight that the efficacy of BTK inhibition depends on disease-specific immune architecture and the relative contribution of BTK-dependent pathways. Safety considerations, including hepatotoxicity signals observed in late-stage trials, and differences between individual compounds further complicate clinical development. In this review, we summarize the mechanistic rationale of BTK inhibition, compare key pharmacological properties across BTK inhibitors, and critically assess clinical trial outcomes. We further discuss key challenges, including patient heterogeneity, trial design constraints, the lack of direct comparative studies, and outline future directions such as biomarker-driven patient selection and the development of BTK degraders.
Gadolinium [Gd (III)] released from gadolinium-based contrast agents, commonly used in magnetic resonance imaging, may accumulate in organs, altering metal homeostasis. We aimed to clarify Gd (III), iron and other metal accumulation in the kidney and brain, in the short (2 days) and long term (20 weeks), following exposure to gadoteric acid (Gd-DOTA) or free Gd (III). Wistar rats received one dose of Gd (III), Gd-DOTA, or saline. Gd (III) and metal levels in the blood (whole blood, and serum in the case of iron), kidney, and brain, and iron metabolism biomarkers, were assessed. Gd (III) was detectable in the blood, kidney, and brain, at both time points, for both Gd (III) and Gd-DOTA-treated groups. Exposure to free Gd (III) showed a significant Gd (III) accumulation in the kidney, brain and blood in the short term and long term; Gd-DOTA presented significant accumulation only in the short term. Gd-DOTA appears to have faster elimination and minimal brain deposition. Exposure to free Gd (III), but not to Gd-DOTA, led to disturbances in metal homeostasis and in iron metabolism markers (serum ferritin and transferrin saturation), but did not alter tissue iron levels. In summary, the kidney appears as the primary site of accumulation and retention. Despite the safer profile shown by Gd-DOTA in the long term, our data highlight the importance of clarifying the pathophysiological implications of Gd (III) retention and/or accumulation, particularly in vulnerable conditions and repeated exposures.
Pseudomonas aeruginosa infection remains a major challenge in donor lung management and post-transplant respiratory care, particularly in the setting of antimicrobial resistance. Ex vivo lung perfusion (EVLP) provides an opportunity for localized antimicrobial intervention before transplantation and allows detailed assessment of infection-associated lung injury under controlled conditions. In this study, we evaluated inhaled sphingosine in a porcine EVLP model of acute P. aeruginosa airway contamination using three experimental groups: uninfected controls, infected lungs treated with 0.9% NaCl, and infected lungs treated with sphingosine. This design allowed us to assess both the impact of P. aeruginosa infection on EVLP physiology and the effects of sphingosine on bacterial burden and infection-associated lung changes. Infected porcine lungs showed greater declines in dynamic and static compliance than uninfected controls, whereas pulmonary artery pressure, peak airway pressure, oxygen-exchange capacity, lactate accumulation, lung weight gain, and histological injury scores were not significantly worsened under the present experimental conditions. Inhaled sphingosine reduced bronchial P. aeruginosa CFU counts, whereas 0.9% NaCl did not. Sphingosine treatment did not further impair lung physiology, oxygenation, lactate accumulation, histological injury, or lung weight gain. Mechanistic analyses showed association of sphingosine with bacterial cardiolipin and increased colocalization of sphingosine with P. aeruginosa, consistent with a membrane-associated antibacterial mechanism described in previous studies. In exploratory experiments using four explanted human recipient lungs, sphingosine inhalation increased sphingosine levels in bronchial and parenchymal tissue and altered related sphingolipid metabolites, including sphingosine-1-phosphate, ceramide, and sphingomyelin, without apparent acute histological damage in assessed airway samples. One of the four human specimens showed bacterial colonization, in which a marked reduction in detectable bacterial growth after sphingosine inhalation was observed as a hypothesis-generating finding. Together, these data suggest that inhaled sphingosine can reduce acutely accessible bronchial P. aeruginosa burden during porcine EVLP without detectable short-term adverse effects and provide a basis for further investigation of sphingosine-related lipid metabolism and local antimicrobial strategies during EVLP.