IntroductionThe prevention of colorectal cancer (CRC) and its possible relapse remains a daunting challenge due to the unclear mechanisms governing gut dysbiosis and its interaction with the gut–immune axis. The present study aimed to investigate the preventive role of a polyherbal immunostimulatory formulation to restoring microbial diversity in gut and preventive management of colon cancer.MethodsWe developed a novel polyherbal and probiotic-based immunostimulatory formulation with four variants (F3A, F3B, F3C, and F3D), incorporating botanical extracts and probiotics. Initially, plant-derived biological lead compounds were identified using LC/MS/MS. Next, the identified biological leads were subjected to in-silico molecular docking and molecular dynamics simulations with apoptotic biomarker, oncogenic marker, and immunomodulatory biomarkers (i.e., BCL2/BAX, KRAS, BRAF, IL-6, and IL-10), respectively. Followed by, the cellular cytotoxicity of the formulations were analyzed in murine colon cancer cell line (CT26), human colorectal adenocarcinoma cell line (HT29), and normal human embryonic kidney cells (HEK-293). All four formulations were tested for their immunomodulatory potential by western blot analysis in in CT26 cell line. Furthermore, E. coli growth inhibition, and T-cell and B-cell proliferation were examined by ex-vivo conditions.ResultsFormulation F3B showed maximal cytotoxicity in CT26 cell line however it considered for animal experiments. Thereafter, in-vivo experiments were conducted using adult female BALB/c mice to investigate the modulation of gut dysbiosis. In the present study, animals were divided in three groups (normal control, diseases control, treatment control) while each group consisted of n = 6 animals.DiscussionOur findings demonstrate that the proposed formulations exhibit anti-proliferative, mild pro-apoptotic, immunomodulatory, and antimicrobial effect. Based on 16S rRNA sequencing results, F3B demonstrated better outcomes toward managing gut dysbiosis in female BALB/C mice. However, we can conclude that formula 3FB can be used as preventive approach to control gut dysbiosis and possible colon cancer relapse.

The present study aims to identify skeletal muscle-specific (α2β2γ1) AMP-activated protein kinase (AMPK) activators by utilizing bioinformatic tools. This study introduces a sensible computational strategy to systematically identify skeletal muscle-specific AMPK activators, thus providing a platform for developing novel therapeutics for Type-2 diabetes mellitus. Owing to the concerning systemic complications and off-target toxicity of the existing anti-diabetic drugs, our preliminary attempt involves structure-based virtual screening of a library of small molecules against the target using a combination of computational tools to reliably pinpoint molecules that could selectively interact with skeletal muscle-specific AMPK complex. The computationally screened library of selective small molecules was retrieved from the PubChem database against the crystal structure of full-length human α2β2γ1 AMPK complex (PDB ID: 6B2E with a resolution of 3.80 Å) from protein databank. The screening comprised a series of approaches, including molecular docking, molecular dynamics simulations, molecular mechanics/generalised born surface area, density functional theory calculations, pharmacokinetics, and toxicity prediction performed using a combination of relevant computational tools. In conclusion, this study has unveiled some interesting in silico hits that seem to have the potential to facilitate the development of a variety of plausible candidate skeletal muscle-specific (α2β2γ1) AMPK activators.

Oral squamous cell carcinoma (OSCC) is the most common aggressive malignancy of the head and neck squamous cell carcinoma (HNSCC) subtype. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression by target degradation or inhibition of translation. miR-21-5p is a multifaceted miRNA found to be overexpressed and acts as a potent oncogene, regulating various cellular pathophysiology, such as cell proliferation, invasion, migration, and apoptosis, in many cancers, including OSCC. Despite its established involvement in OSCC progression, the upstream regulators of miR-21-5p and its downstream targets are still not completely elucidated. Transforming growth factor-beta (TGF-β) is a cytokine that shows a paradoxical role in diverse diseases, including OSCC. TGF-β exerts oncogenic effects in OSCC, shown by the induction of mesenchymal markers, which may account for the metastatic potential of OSCC. Through small RNA sequencing of SCC-25 cells treated with TGF-β, we identified numerous miRNAs that were induced in OSCC. Further, this study confirms the positive correlation between the TGF-β pathway and miR-21-5p induction, in which miR-21-5p is transcriptionally induced by the SMAD-mediated TGF-β signaling pathway. Mechanistically, TGF-β-induced miR-21-5p exerts its oncogenic effects in OSCC by post-transcriptionally suppressing its target, the transcription factor KLF5 (Kruppel-like factor 5). Loss-of-function studies on KLF5 confirmed its tumor suppressive role in SCC-25 and SCC-9 OSCC cells. Taken together, our study reports for the first time the existence of a TGF-β/miR-21-5p/KLF5 regulatory axis in OSCC, which could potentially be of therapeutic value. This article aligns with SDG 3 (Good Health and Well-Being) of the UN Agenda for Sustainable Development.

Recent advancements in nanotherapeutics have revolutionized cancer treatment through the integration of diagnostic and therapeutic modalities, known as theranostics. This critical review examines the current landscape of nanotherapeutics for various cancers, such as bladder and head and neck squamous cell carcinoma, highlighting current advancements in nanotherapeutics and challenges. Key approaches discussed include biomimetic smart nanocarriers, polymeric smart nanocarriers, inorganic-based smart nanocarriers, and nanorobots. Furthermore, diverse nanomaterials have been explored in theranostics, including liposomes, polymeric nanoparticles, and inorganic nanoparticles such as quantum dots and mesoporous silica nanoparticles. Furthermore, the integration of imaging techniques such as surface-enhanced Raman scattering (SERS) and positron emission tomography (PET) with therapeutic nanoparticles has been analyzed for potential clinical applications.

Gut microbiota dysbiosis is increasingly viewed as a disruption of microbial metabolic functions rather than only a shift in microbial composition. Microbiota-derived metabolites not only shape microbial ecology but also directly influence surrounding host tissues by modulating epithelial cell signaling, inflammation, and tumor-associated processes. Among dysbiosis-associated microbes, Fusobacterium is consistently enriched in colorectal cancer (CRC) and contributes to tumor progression, yet the ecological factors regulating its expansion and interaction with host tissues remain unclear. Here, we investigated how microbiome-derived metabolite environments associated with healthy and dysbiotic microbial communities influence Fusobacterium fitness and colorectal epithelial cell behavior. CRC-associated dysbiosis was generated using an orthotopic murine CRC model combined with antibiotic-induced microbiota perturbation. Gut microbial communities were profiled using 16 S rRNA gene sequencing. Metabolite-enriched supernatants derived from healthy gut microbiota, oral microbiota, dysbiotic microbiota and probiotic cultures were evaluated for their effects on CRC-associated bacteria and Fusobacterium sp. growth, adhesion and invasion. These metabolite supernatants were applied to colorectal cancer cells and their effects on viability (MTT assay), migration (scratch assay), apoptosis (Annexin V-FITC flow cytometry), and inflammatory signaling (Western blot analysis of inflammatory markers) were evaluated. CRC-associated dysbiosis showed reduced microbial diversity with enrichment of opportunistic taxa including Fusobacterium and depletion of beneficial commensals such as Lactobacillus and Bifidobacterium. Metabolite-enriched supernatants from healthy gut and oral microbiota suppressed Fusobacterium growth by 55–65

Aggressive Oral squamous cell carcinoma (OSCC) remains a major therapeutic challenge due to its high metastatic potential. OSCC with mutated TP53 often exhibits non-functional or gain-of-function p53 variants, elevated anti-apoptotic protein expression and a strongly anti-inflammatory tumor microenvironment. Although tumor regression using folate receptor (FR)-targeted doxorubicin (DOX) conjugated carbon nanospheres has been reported in syngeneic mouse models, the underlying mechanism remained unexplored. In this study, we introduced a novel dual-targeting approach that simultaneously directs therapy toward the tumor and tumor-associated macrophages (TAMs) via FR-mediated delivery. Compared to pristine DOX, FR-targeted delivery resulted in tumor regression and enhanced drug uptake, particularly within the TAM population. Flow cytometry revealed increased CD80 expression with minimal change in CD163, indicating a modest TAM polarization shift toward an anti-tumoral state. A higher CD80/CD163 ratio in FR-targeted groups suggested potential to overcome TAM-linked immune evasion. Collectively, these findings highlight the novel FR-mediated dual-targeting strategy of CSPs as a promising approach to modulate both the tumor and tumor-associated macrophages for improved chemotherapeutic efficacy in aggressive OSCC. This article aligns with SDG-3 (Good Health and Well-Being) of the UN Agenda for Sustainable Development.

Breast cancer recurrence and therapeutic resistance are primarily driven by breast cancer stem cells (bCSCs), which are poorly eliminated by conventional chemotherapy. To address this limitation, we developed an in situ-forming injectable silk hydrogel for sustained dual delivery of doxorubicin (DOX) and salinomycin (SAL), enabling concurrent targeting of bulk tumor cells and bCSCs. The hydrogel forms under physiological conditions without the need for UV irradiation or toxic initiators and exhibits tunable mechanical and degradation properties. An optimized 8% hydrogel exhibited controlled release behavior of DOX and SAL, characterized by a limited initial burst followed by sustained release for up to 30 days. In vitro viability assays demonstrated that DOX or SAL alone reduced cancer cell viability by approximately 45-55%, whereas the dual-drug loaded hydrogel (SilkVS-DOX + SAL) achieved a cytotoxicity of roughly 90%. Mammosphere formation assays demonstrated a marked reduction in both mammosphere number and size, indicating potent inhibition of bCSC self-renewal. Consistently, expression of bCSC-associated stemness markers was reduced by approximately three-fold relative to free-drug treatments. Flow cytometric analysis further confirmed enhanced induction of apoptosis within the bCSC-enriched population following treatment with SilkVS-DOX + SAL. In vivo studies using 4T1 tumor-bearing mice demonstrated that the localized, sustained release of DOX and SAL from the injectable silk hydrogel synergistically suppressed tumor growth, while significantly reducing systemic toxicity compared to individual drug administration. Overall, the in situ silk injectable hydrogel with sustained dual-drug release effectively eliminates both bulk tumor cells and bCSCs, making this injectable silk hydrogel a promising strategy for reducing breast cancer stem cells, which may reduce the chances of recurrence and enhance therapeutic outcomes.

MicroRNAs (miRNAs) are non-coding regulatory RNAs that are small, single-stranded, and regulate gene expression post-transcriptionally. Mechanistically, miRNAs regulate crucial biological processes, and their dysregulation often contributes to a plethora of diseases, including cancer. miR-21 is one of the major oncomiRs with well-documented aberrant expression in several cancers, including head and neck squamous cell carcinoma (HNSCC). Oncogenic effects of miR-21 are reflected in its regulation of various tumor-suppressor genes and key signaling pathways. In HNSCC, overexpression of miR-21 promotes tumor proliferation, invasion, migration, and resistance to therapy, resulting in recurrence and poor prognosis. Recent studies have revealed role of long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) in regulating miR-21, thereby expanding our understanding of its biological functions. With advances in RNA-based targeted therapeutics, miR-21 has emerged as a potential biomarker, particularly in HNSCC, where early detection and effective intervention remain major challenges. However, there is a lack of focused reviews on the miR-21-lncRNA/circRNA regulatory axis and multiple downstream targets of miR-21 summarized specifically in the context of HNSCC. This review explicitly explores the oncogenic role of miR-21 in HNSCC, its regulation by the lncRNAs and circRNAs axis, its downstream targets, and its potential clinical applications in the diagnosis and treatment of HNSCC.

In this study, we present a hybrid analytical method for the quantification of glimepiride (GLP) from blood plasma and pharmaceutical formulations. Shimadzu HPLC (Model SIL 20 AC HT), a photodiode array detector, and a Phenomenex C18 column (150 × 4.6 mm, 4 μm) were used for method development. A further quality by design (QbD) surface optimization model was applied, and the best-optimized method with chromatographic conditions, such as an injection volume of 15 μl, a column oven temperature of 40°C, a sample cooler temperature of 15°C ± 1°C, a run time of 5 minutes, and a flow rate of 0.8 ml/minute with a mobile phase composition (acetate buffer: acetonitrile, 40:60 ratio), was identified. Finally, the method was validated, and GLP was quantified from various pharmaceutical dosage forms and blood plasma. The results were observed in subsequent laboratories in diluent media and mouse plasma with limits of detection of 0.066 μg/ml and 0.193 μg/ml, respectively, followed by limits of quantification of 0.199 μg/ml and 0.583 μg/ml, respectively. Subsequently, linearity (r2) was observed at 0.999 for both samples. The AUC was 228 ± 2 nm, and the retention time (RT) was 2.8 ± 0.28 minutes. The plasma matrix effect was calculated to be 81.9%. The research findings revealed that the proposed analytical method could be used for both analytical and bioanalytical applications at the industrial scale.

Aim: This study aims to enhance Glimepiride's (GMP) solubility and transdermal permeability by developing a matrix-type transdermal patch to improve systemic bioavailability, circumvent first-pass metabolism and decrease dosing frequency. Materials and Methods: Matrix-type transdermal patches were developed using the solvent-casting method. Initially, formulations were prepared with varying concentrations of polymers and GMP. Formulations were optimized using a quality-by-design approach using response surface methodology (Box-Behnken Design) via Design of Expert (DoE) software, version 8.0.4. Final formulations included Glimepiride in two forms: (a) solid dispersions of GMP (F4) and (b) pure GMP (F7). These formulations were characterized using various analytical techniques. Quantification of Glimepiride from the Transdermal Drug Delivery System (TDS) patches was conducted using the HPLC technique. Results: In vivo experiments such as hypoglycaemic effect, Skin sensitization and irritations test were performed on adult C57BL6/J mice. Other hand, the in vitro drug release to be fond 99.7 +/- 0.99 % and 93.7 +/- 1.2 % respectively. Similarly, permeability rates for patch (F4 and F7) of 0.141 +/- 0.02 and 0.120 +/- 0.04 mg/cm2/hr were recorded respectively.The results demonstrated that the solid dispersion formulation of GMP (F4) exhibited superior permeation and physicochemical properties compared to the pure GMP formulation (F7). Conclusion: In conclusion, the proposed transdermal formulation may serve as an alternative to solid oral formulations, effectively bypassing first-pass metabolism and minimizing the frequency of dosage administration.

Amyotrophic Lateral Sclerosis (ALS) is a progressive neurodegenerative disorder affecting nerve cells in the brain and spinal cord. With a global incidence of 1.9 to 6 per 100,000 people, ALS is slightly more common in men and prevalent in individuals over 60. However, this review provides a concise update on the regulatory landscape and therapeutic advancements in managing ALS, focusing on the recent approval of Tofersen, the first gene therapy specifically targeting SOD1 mutation-related ALS. It highlights Tofersen unique role as an orphan drug approved by the US FDA, emphasizing its mechanism of action, gene silencing and its impact on reducing neurodegeneration. Additionally, the review synthesizes data from ongoing clinical trials, pharmacovigilance reports, and case studies to comprehensively understand Tofersen’s safety, efficacy and market exclusivity. Beyond this, it explores the emerging potential of nanotherapeutic approaches to ALS treatment, identifying critical research gaps and future directions. Integrating regulatory updates, clinical evidence, and innovative therapeutic strategies, the review uniquely contributes to the ALS literature by bridging current treatment realities with potential future therapies, aiming to inform researchers, clinicians, and policymakers on optimizing ALS management.

Targeting glucocorticoid Receptors (GR) induces gluconeogenesis in cancer cells, potentially disrupting their glycolytic dependency and acidic tumor microenvironment (TME), thereby creating an energetically unfavourable state and reducing drug resistance by impairing the acid reflux mechanism. Based on this rationale, we developed a GR-mediated liposomal co-delivery system, D1XP-p53, carrying the tumor suppressor gene, p53, and the chemotherapeutic drug, paclitaxel, to overcome the limitations of conventional anti-cancer therapies and to assess whether wild-type p53 enhances the anti-cancer activity of paclitaxel against Oral Squamous Cell Carcinoma (OSCC).In vitrostudies demonstrated that D1XP-p53 selectively decreased the viability of OSCC cells and significantly inhibited their migration, invasion, and proliferation. Mechanistic investigations revealed an upregulation of the BAX/BCL2 ratio when oral cancer cells were treated with D1XP-p53, indicating the activation of intrinsic apoptotic pathways. The efficacy of D1XP-p53 was further validated in 3D spheroid models using MOC2 and FaDu cell lines, where it significantly reduced spheroid-forming ability and upregulated E-cadherin expression, indicating its potential role in enhancing anti-cancer activity and mitigating cellular migration.In vivoexperiments using a murine model of OSCC with MOC2 cells showed a marked reduction in tumor volume in mice treated with D1XP-p53, with minimal systemic toxicity as assessed by H&E staining and biodistribution analysis. Considering the crucial role of TME components such as tumor-associated macrophages, cancer stem cells, and growth factors in tumor progression and metastasis, we further evaluated the impact of our delivery system, D1XP-p53, on these elements. We observed that D1XP-p53 treatment in mice significantly upregulated the M1/M2 ratios and decreased thec-mycandSOX2expression, indicating the potential role of the delivery system in modulating the TME components. These findings collectively demonstrate that the GR-targeted co-delivery system, D1XP-p53, enhances anti-cancer activity and modulates the TME, offering a promising multi-modal treatment against aggressive oral cancer.

This chapter aim to explore transformative role of artificial intelligence (AI) in detecting and diagnosis of cancer, addressing critical gaps in early detection, diagnosis, personalized treatment, and improved patient outcomes. AI based algorithms, including machine learning and deep learning, are increasingly utilized to analysed complex microbiome data, identify cancer-associated signature, and predict therapeutics response. Moreover, we elucidate how AI facilitates the integration of multi-omics data (genomics, transcriptomics and metabolomics) with microbiome profiles, enhancing our understanding of cancer pathogenesis and progression. It also examines the application of AI in developing non-invasive diagnostic tools using microbiome biomarkers for various cancer, such as colorectal, lung, and breast cancer. Furthermore, the chapter addresses the challenges and opportunities in leveraging AI for microbiome-based caner immunotherapy. Conclusively, by highlighting future directions and the potential of AI revolutionize cancer care through microbiome-based precision medicine.

Cancer stem cells (CSCs) are a crucial subpopulation in gynecological tumors, defined by their self-renewal, differentiation potential, and resistance to conventional therapies. These cells are central to tumor initiation, progression, metastasis, and recurrence, making them key targets for innovative therapeutic strategies. This chapter will explore the molecular mechanisms that regulate CSCs, focusing on signaling pathways such as Wnt, Notch, and Hedgehog, which are critical for CSC maintenance and survival. It will also examine emerging therapeutic approaches aimed at eradicating CSCs, including pathway inhibitors, immune-based strategies, and combinatorial treatments. By targeting CSCs, these approaches hold the promise of overcoming resistance and achieving more reliable clinical responses in gynecological cancers. Further, this chapter delves into the challenges and future directions of translating CSC-targeted therapies into clinical practice.

This review explores the impacts of synthetic pesticides and biopesticides on human health, aiming to provide a comprehensive understanding of their benefits and risks. Currently, farmers are using synthetic pesticides to increase the yield of crop production, but they pose significant health risks, such as acute poisoning, cancer, endocrine disruption, anaphylactic shock, and other severe health issues. On the other hand, biopesticides, derived from natural organisms or plant-derived secondary metabolites, are considered safer alternatives, offering effective pest management with reduced risk to human health. This review draws attention to plant-derived materials in pest control management. Further, deciphering plant diseases with phytogenic bacteria and their control by organic bio-pesticides. Conclusively, this review suggests that future research should focus on integrated pest management approaches that combine the strengths of both synthetic and biopesticide applications while mitigating health risks. The findings underscore the imperative for ongoing evaluation of pesticide usage and provide a framework for informed decision-making regarding human exposure to these substances.

Gelatin hydrogels have drawn attention for their diverse biomedical applications due to their flexible physiochemical properties. However, such gelatin hydrogels are made of toxic crosslinkers and photoinitiators, restricting their non-invasive deep tissue application. The in-situ forming chemical crosslinked without such toxic crosslinker and UV light has not been explored under physiological conditions. This study establishes a simple method to fabricate an injectable click-chemistry-based in-situ forming gelatin hydrogel in a physiological environment (without toxic UV or photoinitiator) with tunable physiochemical properties to modulate cellular response. Using Divinyl Sulfone (DVS) modification, gelatin hydrogel (GelVS) is optimized with tunable degradation properties, moduli (100 Pa -1000 Pa), gelation time, swelling, degradation, and viscoelastic behaviour. The in-vitro results using fibroblast and stem cells show that the hydrogel and its precursors were cytocompatible with diverging feedback of cells as the modulus varies. The in-vivo analysis for injectability, degradation, and biocompatibility of the GelVS hydrogel displays their biocompatible nature and lasts up to 30 days at the injecting site. Overall results indicate that DVS-modified GelVS hydrogel will be a great system with tunable physicochemical properties to modulate favorable cellular response for tissue regeneration and noninvasive deep tissue application.

STAT3 is an important protein responsible for cellular proliferation, motility, and immune tolerance and is hyperactive in colorectal cancer, instigating metastasis, cellular proliferation, migration, as well as inhibition. It helps in proliferation of myeloid-derived suppressor cells (MDSCs), which within the tumor microenvironment (TME) suppress T cells to encourage tumor growth, metastasis, and resistance to immunotherapy, besides playing dynamic role in regulating macrophages within the tumor. Thus, MDSC is a potential target to augment immune surveillance within the TME. Herein, we report targeting both colorectal cancer and MDSCs using a glucocorticoid receptor (GR)-targeted nanoliposomal formulation carrying GR-ligand, dexamethasone (Dex), and a STAT3 inhibitor, niclosamide (N). Our main objective was to selectively inhibit STAT3, the key immunomodulatory factor in most TME-associated cells including MDSCs, and also repurpose the use of this antihelminthic, low-cost drug N for cancer treatment. The resultant formulation D1XN exhibited better tumor regression and survivability compared to GR nontargeted formulation. Further, bone marrow cell-derived MDSCs were engineered by D1XN treatment ex vivo and were inoculated back to tumor-bearing mice. Significant tumor growth inhibition with enhanced antiproliferative immune cell signatures, such as T cell infiltration, decrease in Treg cells, and increased M1/M2 macrophage ratio within the TME were observed. This reveals the effectiveness of engineered MDSCs to modulate tumor surveillance besides reversing the aggressiveness of the tumor. Therefore, D1XN and D1XN-mediated engineered MDSCs alone or in combination can be considered as potent selective chemo-immunotherapeutic nanoliposomal agent(s) against colorectal cancer.

Glucocorticoid and Mineralocorticoid receptors are principally ligand-dependent intracellular transcription factors that are known to influence the development and growth of many human cancers. Our study investigates the potential of these receptors to act as a target for oral cancer treatment since findings in this regard are sparse till date. Leveraging the aberrant behavior of steroid hormone receptors (SHRs) in cancer, we have targeted oral cancer cells in 2D-culture using liposomes containing both synthetic as well as crude, natural SHR ligands isolated from an aqueous Indian medicinal plant. Lipoplexes thus formulated demonstrated targeted transfectability as indicated by expression of green fluorescent protein. Transfection of oral squamous cell carcinoma cells with exogenous, anticancer gene p53 lipoplexed with crude saponin-based liposome induced apoptosis of cancer cells via regulation of BAX and B-cell leukemia/lymphoma-2 (BCL2) protein levels at levels comparable with pre-established delivery systems based on synthetic SHR ligands. Our findings strongly indicate a possibility of developing plant saponin-based inexpensive delivery systems which would target cancer cells selectively with reduced risks of off target delivery and its side effects.

N6-methyladenosine (m6A) plays a pivotal role in regulating epitranscriptomic mechanisms and is closely linked to the normal functioning of diverse classes of RNAs, both coding as well as noncoding. Recent research highlights the role of m6A RNA methylation in the onset and progression of several cancers, including head and neck squamous cell carcinoma (HNSCC). HNSCC ranks as the seventh most common cancer globally, with a five-year patient survival rate of just 50