Dysregulation of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors underlies a variety of neurological conditions, including epilepsy and neurodegenerative disorders. Pharmacological approaches that selectively modulate AMPA receptor subunits offer significant promise in mitigating glutamate-mediated excitotoxicity. We investigated eight newly synthesized coumarin–benzodiazepine (CD) hybrid derivatives designed as negative allosteric modulators of AMPA receptors, seeking both potency and selectivity. Using whole-cell patch-clamp electrophysiology in HEK293 cells expressing specific AMPA receptor subunits (GluA1–4), we evaluated the ability of each compound to alter receptor kinetics, particularly desensitization and deactivation. Among the tested derivatives, one molecule (CD8) exhibited a notably high affinity for AMPA receptors, with potent inhibition observed across multiple subunit combinations. Crucially, CD8 displayed minimal off-target activity against kainate and NMDA receptors, underscoring its selectivity for AMPA receptor subtypes. Structurally, CD8’s carboxyl substituent at the para position of the phenyl ring emerged as pivotal for enhanced receptor binding and negative modulation. Furthermore, kinetics analyses revealed that certain derivatives, including CD8, not only attenuated peak current amplitude but also accelerated deactivation rates and reduced desensitization—indicative of a robust negative allosteric mechanism distinct from the orthosteric glutamate-binding site. Taken together, these findings highlight the potential of coumarin–benzodiazepine hybrids as selective modulators of AMPA receptors. By dampening hyperexcitatory neuronal signaling through targeted receptor inhibition, these derivatives hold promise as a therapeutic avenue for a range of central nervous system pathologies. Future research will probe how these compounds interact with auxiliary proteins (e.g., TARPs) to further optimize AMPA receptor gating in native neuronal environments, thereby refining their translational potential for neurological conditions associated with abnormal glutamatergic transmission.

Introduction

Wiskott--Aldrich Syndrome patients feature defects in T cell activation and proliferation, but also in megakaryopoiesis and platelets. We discovered that diacylglycerol kinase alpha (DGKα) constrains T-cell activation, and its activity is limited by a pathway comprising SAP (an adaptor mutated in Duncan’s disease) and the WAS protein (WASp). We propose that, in the context of SAP or WASp deficiency, there is excessive DGKα activity contributing to diseases phenotypes.

Methods

In T cells, we triggered immune synapse (IS) formation among Jurkat triple reporter and superantigen-loaded Raji cells. Jurkat triple reporter cells allow for the quantification of NFAT, NFκB, and AP-1 activity induction using flow cytometry.

Similarly, we silenced WAS in the erythroleukemia cell line (HEL) and induced differentiation through PMA treatment measuring cell elongation, spread, and CD41 induction.

Results

IS analysis confirmed that SAP- and WASp-deficient cells have a reduced capacity to form IS and, surprisingly, DGKα inhibition further reduces it, underling an important role of DGKα in shaping IS. SAP- and WASp-deficient cells also show a defect in NFAT and AP-1 signaling pathways, whereas NFκB activity is unaffected. DGKα inhibition rescues both NFAT and AP-1 defects but does not interfere with NFκB.

In megakaryocytes, DGKα dampers thrombopoiesis and platelet aggregation. DGKα inhibition rescued the elongation defect of WASp-deficient HEL cells, but was not able to restore CD41 induction.

Conclusions

Thus, in the absence of SAP or WASp, excessive DGKα activity consumes diacylglycerol, perturbing signaling. Indeed, DGKα inhibition is required for NFAT and AP-1 activation, and this can be compensated by DGK inhibitors.

Similarly, excessive megakaryocyte DGKα activity in WAS reduces cytoskeletal remodelling, and this is also restored by DGK inhibitors.

All of these observations suggest that inhibiting DGKα activity could represents a novel therapeutic approach for WAS.

Over time, advancements in cancer diagnostics and therapies have been remarkable. This study compares normal (CCD-18Co) and cancerous (CaCo-2) cell lines from the human gastrointestinal tract, analyzing their nanomechanical and biochemical properties to identify potential cancer biomarkers that could enhance oncological diagnostics. The research examines the effects of 24 and 48 hours of 10uM supplementation with statins—mevastatin, lovastatin, and simvastatin—on the biochemical and nanomechanical properties of these cells, employing Raman imaging and AFM techniques. The innovative integration of these techniques provides a comprehensive biochemical and nanomechanical characterization of normal, cancerous, and statin-treated cancer cells, highlighting how different statins influence cellular properties.

A primary objective of this study was to identify statistically significant differences between normal and cancerous colon cells, including cancer cells treated with mevastatin, simvastatin, and lovastatin, based on their vibrational features. We systematically explored how Raman imaging and spectroscopy respond to normal and cancerous human cells, both untreated and treated with statins. Our approach demonstrates a fast, cost-effective method for visualizing cellular structures and performing virtual staining. By incorporating Raman intensity into pseudo-color images, these high-resolution, label-free images allow direct analysis of cellular substructures, facilitating the detection of biochemical changes characteristic of cancer progression and aiding in the assessment of anti-cancer therapies.

Using Raman data, we analyzed and compared the vibrational features of normal, cancerous, and statin-treated cancer cells. Average spectra and Raman band intensity ratios for key biological components—proteins, nucleic acids, and lipids—enabled detailed insights into the molecular changes associated with cancer and its treatment.

Our findings provide evidence supporting the hypothesis that long-term use of lipophilic statins may influence cancer incidence, including specific cancer types. Additionally, statins may enhance chemotherapy sensitivity and improve clinical outcomes for patients already diagnosed with cancer.

Colorectal cancer is the second most prevalent cancer worldwide, while gastric cancer shows an increasing frequency, especially in young people, accounting for over one million new patients in 2022. This study investigates the genetic associations between colorectal and gastric cancer.

We conducted a systematic review focusing on the genetic pathways associated with colorectal and gastric cancer. Articles were retrieved from the PubMed database. The inclusion criteria encompassed the keywords “genetics”, “colorectal cancer” and “gastric cancer” in papers published between 2023 and 2024. The exclusion criteria were case report papers.

A total of 56 articles met the eligibility criteria. Of these, eight articles explored Li–Fraumeni and Lynch syndromes, which are linked to a high risk of developing colorectal and gastric tumors due to mutations in the TP53 and MMR genes, respectively. Furthermore, 11 studies emphasized the elevated risk of malignancy due to gastrointestinal polyps in familial adenomatous polyposis (APC mutations); juvenile polyposis syndrome (SMAD4/BMPR1A mutations); and Peutz–Jeghers syndrome (STK11 mutations). We identified five studies that linked inflammatory bowel disease to colitis-associated colorectal carcinoma, driven by p53 mutations. Furthermore, three papers explored gastrointestinal stromal tumors, rare neoplasms that occur due to PDGFRA D842V mutations. Additionally, 29 articles showed that genetic mutations in the Wnt Signalling Pathway and KRAS gene, polymorphisms in nucleic acids and chemokine ligands promote carcinogenesis, tumor progression and invasion, indicating a higher risk of developing metastasis. One of the most relevant targets for developing new therapies is the Wnt receptor ROR1, which is overexpressed in several cancers and promotes Wnt Signalling Pathway hyperactivation, facilitating carcinogenesis.

Our findings suggest that genetic exploration serves as a predictive factor for the co-occurrence of colorectal and gastric cancer. Mutations in genes linked to genetic syndromes and signalling pathways are associated with an increased risk of gastrointestinal tumorigenesis, particularly involving the interplay between colorectal and gastric tumors.

Objective: Human papillomavirus (HPV) infection is a significant risk factor for head and neck squamous-cell carcinoma (HNSCC). This study aims to identify specific signaling pathways, biomarkers, and potential targeted drugs for HPV-positive HNSCC through competitive endogenous RNA (ceRNA) network analysis.

Methods: High-throughput transcriptomic sequencing was performed on UPCI:SCC154 (HPV-positive HNSCC), Cal27 (HPV-negative HNSCC), and normal human oral keratinocytes to identify differentially expressed mRNAs and miRNAs associated with HPV-positive HNSCC. The interactions of competitive endogenous RNAs were predicted using TargetScan and miRanda, and protein–protein interactions were predicted using the String database to construct a ceRNA network. Network topology analysis was employed to identify hub genes in HPV-positive HNSCC. Further, upstream miRNAs of the hub genes were screened by integrating multiple databases, including encori, mircode, miRDB, miRTarBase, miRWalk, and picTAR, to construct miRNA-mRNA signaling axes. Drug repositioning analysis was conducted using the Connectivity Map database to predict small-molecule drugs targeting the hub genes. Molecular docking and dynamics simulations were used to evaluate the binding affinity and stability of the small molecules with the target proteins. The efficacy of the drugs was validated through cellular functional assays.

Results: A total of 3,253 differentially expressed mRNAs and 391 miRNAs were identified in HPV-positive HNSCC, and RPAP2 was identified as a hub gene. Integrated multi-database analysis revealed that hsa-miR-181b-3p potentially targets and regulates RPAP2. Drug repositioning screening identified five potential drugs, among which molecular docking and dynamics simulations indicated that Enzastaurin exhibited excellent binding affinity and stability with RPAP2. Cellular experiments demonstrated that, compared to cisplatin, Enzastaurin significantly reduced the viability of HPV-positive HNSCC cells and suppressed the expression levels of RPAP2 and hsa-miR-181b-3p.

Conclusion: The miR-181b-3p/RPAP2 axis plays a key regulatory role in HPV-positive HNSCC. Enzastaurin demonstrates promising therapeutic potential, offering a foundation for targeted drug development.

In skin cancer treatment, the often severe side effects and limited efficacy of immunotherapies have prompted the development of novel therapeutic strategies. Hence, innovative formulations using nanosystems have emerged, which, due to having a nanometric size, being able to encapsulate and protect loaded drug molecules, leading to controlled drug release, providing increased permeation, and having tunable properties, have proven to improve therapeutic outcomes. Among the many types of nanoplatforms, nanoemulsions and nanoemugels have been some of the most successfully developed for topical immunomodulatory skin cancer treatment, including nanoemulgels, combining the immunomodulatory effects of imiquimod and the anti-inflammatory properties of curcumin, and nanoemulsions containing plant-derived anticarcinogenic and immunosuppressive molecule triptolide, which have led to increased skin drug permeation and retention and low toxicity in tissue models. On the other hand, drug repurposing, making use of already marketed drug molecules and repositioning them for antitumor effects, has also been described for cancer immunotherapy, with doxycycline, a broad-spectrum antibiotic with recognized immunosuppressive and anticancer properties, having been incorporated into multifunctional hybrid electrospun nanofibrous scaffolds containing hydroxyapatite nanoparticles, yielding synergistic anticancer effects in in vitro skin cancer models. Furthermore, dual drug-loaded nanosystems have also proven to be beneficial in tumor immunotherapy, including immunoliposomes, simultaneously encapsulating the chemotherapeutic agent 5-fluorouracil and the antibody cetuximab, or immunoliposomes co-loaded with the immunomodulating compound bufalin and the anti-CD40 antibody, revealing increased and synergistic anticancer effects in in vitro and in vivo models of skin cancer, with or without the added use of iontophoresis, resulting in increased cellular uptake via endocytosis mediated by antigen–antibody interaction, simultaneous and long-lasting antigen delivery, with the induction of tumor apoptosis, and decreased tumor volume and tumor weight. Therefore, the use of nanosystems, drug repurposing, and synergistic dual drug-loading might be key to ensuring the success of future skin cancer immunotherapies.

Introduction: Type 2 diabetes mellitus (T2DM) affects 462 million people globally and is linked to complications like retinopathy, nephropathy, neuropathy, and coronary artery disease (CAD). T2DM increases CAD risk, contributing to 75% of related mortality. Genetic predispositions and distinct mechanisms differentiate DM-CAD from other forms, such as CAD, caused by hypertension or degenerative changes, requiring gene expression profiling and transcriptome and reactome analyses to identify and establish molecular markers and improve its diagnosis, prognosis, and treatment strategies. Methods: The Gene Expression Omnibus (GEO) datasets GSE250283 and GSE90074 were used to identify the differentially expressed genes involved in DM-CAD. Biological targets were identified that coincided with the identified differentially expressed genes. The biological pathways were analyzed using KEGG, and hub genes were identified for a further functional and signaling pathway analysis.

Results: The differentially expressed gene biomarkers were identified between the control and DM and DM-CAD. NLRP3, TLR4, STAT3, IL6, TNF-α, and NF-κB were found to be upregulated, while PPARG, SIRT1, and ADIPOR1 were downregulated, indicating significant pathways with involvement, including the oxidative stress response, JAK-STAT signaling, and insulin resistance and mitochondrial dysfunction.

Conclusion: Oxidative stress emerges as a critical driver of T2DM-CAD pathogenesis, influencing inflammatory pathways and metabolic dysfunction. Genes such as SOD2, CAT, and GPX1 highlight disruptions in antioxidant defense mechanisms, aligning with mitochondrial dysfunction. Elevated STAT3 expression in the JAK-STAT pathway and NLRP3 activation further exacerbate oxidative damage and inflammation. Meanwhile, the downregulation of SIRT1 and ADIPOQ underscores impaired glucose regulation and insulin sensitivity. These findings position oxidative stress as a key therapeutic target, alongside inflammasome and immune signaling pathways like the JAK-STAT pathway as novel therapeutic targets for mitigating T2DM-CAD's severity.

Peroxisome proliferator-activated receptor gamma (PPARγ) is a crucial nuclear receptor in humans, playing a central role in regulating glucose homeostasis and lipid metabolism. Its activation holds significant potential not only for managing Type 2 diabetes but also for preventing its onset by addressing insulin resistance, a key precursor to the disease. This study explores the potential of polyphenols as natural modulators of PPARγ, aiming to enhance strategies for preventing the progression of insulin resistance to Type 2 diabetes. Polyphenols, plant-derived bioactive compounds, possess strong bioactivity in modulating various cellular pathways, including those linked to PPARγ activation. Since insulin resistance can develop years before the onset of Type 2 diabetes, identifying compounds that enhance insulin sensitivity early is critical for disease prevention. Molecular docking studies were conducted on a library of 12,000 polyphenols to identify compounds with the strongest binding affinities for PPARγ. Among the candidates, Resveratrol and Epigallocatechin gallate (EGCG) emerged as top ligands, with docking scores of -9.38 kcal/mol and -8.77 kcal/mol, respectively, along with lower RMSD values (0.55 and 0.72). These compounds demonstrated strong binding to key receptor sites of PPARγ, potentially enhancing its activity and improving insulin sensitivity, thereby reducing the risk of Type 2 diabetes development. Moreover, the activation of the adiponectin signaling pathway was identified as a crucial mechanism underlying the beneficial effects of PPARγ in mitigating insulin resistance. Molecular dynamics simulations (MDSs) further validated the stability and efficacy of polyphenol–PPARγ complexes, confirming their sustained interactions and enhanced receptor activation. The pharmacokinetic profiles of these polyphenols revealed highly favorable characteristics, such as excellent bioavailability and non-toxicity, reinforcing their potential as preventive agents. Collectively, this study provides new insights into the modulation of PPARγ by polyphenols, offering a promising approach for delaying or preventing Type 2 diabetes through natural compounds that target early metabolic dysfunction.

Ovarian cancer is the deadliest gynecological malignancy, with around 310,000 women worldwide being diagnosed with the cancer in 2020. Resistance to chemotherapeutics is inevitable in most ovarian cancer patients, but little is known about the precise mechanisms of chemoresistance in ovarian cancer. This study aimed to uncover novel chemoresistance mechanisms using the bioinformatics tools GEO2R and DAVID by compiling a list of differentially expressed genes and then clustering them into specific expression patterns. Differential gene expression data of in vitro studies comparing paclitaxel- and cisplatin-resistant subtypes to wild-type counterparts were obtained from GSE73935 and GSE26465 and analyzed in GEO2R. A common list of 8,697 differentially expressed genes between both datasets was extracted using the dplyr package in R. The list of common genes was input into the DAVID functional annotation clustering tool. Results were considered statistically significant at the threshold of (p<0.05) and a false discovery rate under .05. The protein–protein interactions network was used in order to ascertain the molecular mechanisms of specific differentially expressed proteins in relation to the clusters they relate to. Genes that were differentially expressed fell into established patterns of chemoresistance well established in ovarian cancer, such as epithelial–mesenchymal transition and autophagy. However, expression profiles consistent with chemoresistance pathways in other cancers, but not well established in ovarian cancer, were also identified, namely pathways of transcription regulation, ubiquitin-binding, protein transport, the TGF-beta signaling pathway, cell projection, and cell polarity. These findings suggest novel chemoresistance pathways that could be targeted to resensitize chemoresistant cells to chemotherapeutics or for early prediction of chemoresistance in patients.

Introduction:

Kalanchoe pinnata has long been recognised for its medicinal properties, especially in the treatment of metabolic diseases like obesity. This study examines the phytochemical makeup of K. pinnata root–stem methanol extract and looks at its potential as a treatment for weight management.

Method:

Using a thorough phytochemical analysis that combined qualitative and quantitative techniques, the bioactive components of the root–stem powder were identified and measured. Functional groups were determined using FTIR and UV–Vis spectroscopies. GC-MS analysis was used to profile volatile chemicals. An in vitro pancreatic lipase inhibition experiment was used to evaluate the methanol extract's anti-obesity efficacy. To find out how the extract affected lipid profiles and histopathological alterations in the pancreas and other organs, an in vivo investigation employing a high-fat diet was also carried out.

Results:

The findings of the phytochemical screening revealed the presence of compounds such as flavonoids, polyphenols, saponins, and glycosaponins, which are known to enhance metabolic health. At the same time, the GC/Mass detected essential compounds, such as different fatty acids and phytol octadecanoic acid. The K. pinnata extract in methanol markedly inhibited pancreatic lipase activity. After the extract treatment and a high-fat diet, the rats' lipid profiles improved and displayed decreased LDL and total cholesterol levels. Restoring the normal pancreatic liver and kidney architecture and reducing inflammation were found by histopathological examinations, and these effects were comparable to those of the orlistat.

Conclusion:

The study's findings demonstrate that K. pinnata has two key anti-obesity benefits: enhanced metabolic parameters and preventive effects on organ health. The results necessitate more research into the precise mechanisms of action and possible therapeutic uses of this plant extract in the treatment of obesity.