Vibhav Gautam

Background: Engineered nanomaterials (NMs) are widely tested for biological safety using standard colorimetric, fluorometric, and enzymatic in vitro assays. However, the unique physicochemical properties of NMs, including strong optical activity, high surface-to-volume ratios, and surface reactivity, create a high potential for systematic, often unrecognized interference with these assays. Objectives: This systematic review sought to i) provide a comprehensive catalogue of reported cases of NM-induced assay interference, ii) quantify interference severity and direction by NM type and assay system, iii) synthesize reported physicochemical characteristics associated with assay interference, and iv) provide evidence-based recommendations for mitigation. Methods: A systematic search of PubMed, Web of Science, and Scopus (2006-2026) identified 52 eligible records for the evidence synthesis, comprising primary experimental or method-validation studies together with contextual evidence. Data were extracted on NM composition, size, surface chemistry, assay type, interference mechanism, and effect magnitude. Interference severity was scored semi-quantitatively (0–3). A systematic narrative synthesis was performed. Results: Interference was documented across multiple nanomaterial categories and assay platforms represented in the evidence matrix, with the extent of evidence varying among individual nanomaterial–assay combinations. Carbon-based nanomaterials demonstrated interference across several commonly used viability assays, although the concentrations and experimental conditions associated with interference varied substantially among studies. Silver NPs exhibited a strong LDH interference due to protein adsorption and ROS-mediated deactivation. The iron oxide NPs showed concentration-dependent optical interference across the experimental spectral ranges examined in the contributing studies. Cytokine ELISA tests were vulnerable to cytokine sequestration by TiO ₂ , SiO ₂ and cationic NPs. Consistent with prior population-level literature surveys, not restricted to this review’s eligibility-filtered cohort, the proportion of nanotoxicology publications omitting interference controls has been reported to fall from over 90% before 2014 to approximately 45% by 2021–2024, underscoring the reporting gap that motivated this review’s interference-control eligibility criterion. Conclusion: NM-induced assay interference is pervasive, NM-specific, assay-specific, and concentration-dependent. No universal correction exists; each NM–assay combination requires individual validation. A standardized five-step interference testing workflow and reporting checklist are proposed to improve reproducibility in nanotoxicology.

Despite significant advances in cancer diagnosis and treatment, cancer remains a major cause of disease burden and mortality worldwide [1]. Its complexity arises from extensive molecular and cellular heterogeneity, which influences tumor initiation, progression, metastatic spread, and treatment response [2]. The rapid evolution of high-throughput OMICS technologies, including genomics, transcriptomics, proteomics, epigenomics, metabolomics, and single-cell profiling, has significantly advanced cancer research by enabling comprehensive characterization of tumors across multiple molecular layers and biological scales [3; 4; 5] Taken together, the six contributions in this research topic underscore the transformative role of integrating robust molecular data with rigorous analytical approaches in advancing precision oncology. Spanning methodological advances in gene expression analysis, populationspecific genomic profiling, proteogenomics, circulating biomarkers, multimodal liquid biopsy, and evidence-based therapeutic strategies, these studies collectively demonstrate how integrative OMICS-based approaches can improve our understanding of cancer biology while accelerating biomarker discovery, improving patient stratification and clinical outcomes. We hope that the work presented in this research topic will stimulate further interdisciplinary collaboration among experimental biologists, computational scientists, and clinicians, fostering the development of next-generation OMICS-driven approaches for cancer diagnosis, prognosis, surveillance and personalized cancer therapy.

Visceral leishmaniasis (VL), a parasitic disease caused by Leishmania donovani , requires a robust CD4 ⁺ T‐cells response to control parasite replication by interferon‐gamma (IFN‐γ) production and activation of macrophages. However, in VL patients, the anti‐parasitic CD4 ⁺ T‐cell responses are ineffective for reasons that are still unclear. Our recent study reporting a transcriptional signature of CD4 ⁺ T‐cell isolated from the peripheral blood of active VL patients showed enhanced expression of genes related to cytotoxicity. This study investigates the cytotoxic potential of CD4 ⁺ T‐cells in VL patients, focusing on the expression of the key cytotoxic molecules granzyme B (GZMB), granulysin (GNLY), perforin (PRF), natural killer cell granule protein 7 (NKG7) and the degranulation capacity of CD4 ⁺ T‐cells during infection. We observed significant upregulation of these cytotoxic markers in CD4⁺ T‐cells from VL patients, particularly prior to anti‐parasitic drug treatment (D0), suggesting activation of these cells. However, the degranulation capacity, as indicated by CD107a expression, was comparable between VL patients and endemic controls (ECs), suggesting potential functional impairment in the cytotoxic response. Yet, antigen‐specific GZMB secretion in whole blood cultures and intracellular GZMB production in CD4 ⁺ T‐cell subsets (notably Th1, Th9 and Th17/22 cells) were enhanced in VL patients, indicating a robust antigen‐specific response, though this did not translate into effective parasite control. These findings highlight a paradox whereby CD4 ⁺ T‐cells from VL patients have heightened production of cytotoxic molecules, but lack translocation of CD107a to the cell surface, which is associated with an inability to eliminate parasites. This study provides new insights into the immune dysfunction in VL, highlighting a potential role for the cytotoxic phenotype that develops in parasite‐specific CD4 ⁺ T‐cells. Targeting these pathways may offer novel therapeutic strategies to enhance immune responses and improve clinical outcomes in VL.

Breast cancer (BC) is a highly complex and heterogeneous malignancy and the most prevalent cancer among women worldwide. The diagnosis, prognosis, and the treatment of BC pose significant challenges that are responsible for their limited therapeutic efficacy. Omics‐based technologies have gained substantial attention in BC diagnosis through molecular profiling and diverse clinical analytics. The integration of metabolomics, proteomics, transcriptomics, and genomics provides a multidimensional approach to personalized BC diagnosis and treatment through high‐throughput molecular profiling. Moreover, the emergence of artificial intelligence (AI) has also supported more accurate and early diagnosis of BC through multimodal integration of diverse datasets. The integration of advanced deep learning (DL) and machine learning (ML) has been extensively exploited for tumor grading, histopathological classification, molecular profiling, diagnostic imaging, and prognostic prediction. This review aims to summarize recent developments in AI‐driven multi‐omics approaches for the discovery of BC biomarkers. We have also highlighted the integration of omics‐based data like metabolomics, proteomics, transcriptomics, and genomics with key AI techniques, including ML and DL, that play a crucial role in the inclusion of multi‐omics in cancer and biomarker discovery. We have further discussed AI‐based BC screening and diagnostic approaches, as well as the contribution of AI models for patient stratification, biomarker discovery, and prediction of therapeutic response. Additionally, key limitations and challenges, including data heterogeneity, high computational complexity, and model interpretability, have also been highlighted in the present review. Conclusively, we have also outlined future perspectives on the integration of AI and multi‐omics to revolutionize precision clinical medicine and improve clinical outcomes in BC theranostics.

The CRISPR-Cas systems, identified initially as adaptive immune mechanisms in bacteria and archaea against viral threats, have rapidly evolved into transformative tools in genetic engineering and biotechnology. These RNA-guided systems are broadly classified into Class 1, comprising multi-subunit complexes, and Class 2, characterized by compact single-effector protein, such as Cas9, Cas12, and Cas13. Their remarkable structural and functional diversity enables microorganisms to adapt to diverse ecological niches, offering a vast repertoire of genome-editing strategies. Beyond their natural role in maintaining genome integrity and defense, CRISPR-Cas systems have been extensively repurposed for precise genome modification, transcriptional regulation, epigenetic editing, and nucleic acid detection. Recent advances in computational mining of microbial genomes and metagenomes have uncovered a broad range of novel CRISPR effectors with unique properties, distinct protospacer adjacent motif (PAM) requirements, RNA-targeting capabilities, miniature architectures, and promiscuous cleavage activities that significantly expand the molecular biology toolkit. The development of CRISPR-based technologies such as base editing, prime editing, gene knock-in/out, and live-cell DNA/RNA imaging exemplifies the versatility of these systems. Despite the challenges associated with delivering complex Class 1 systems, both classes are now being actively harnessed across diverse microbial platforms. Concurrently, the CRISPR-Cas research, particularly for guide RNA (gRNA) design and activity prediction, has revolutionized target specificity and editing efficiency. This review presents a comprehensive overview of CRISPR-Cas system diversity, their genomic landscape in microorganisms, and their cutting-edge biotechnological applications. It also emphasizes the transformative potential of CRISPR in synthetic biology, therapeutics, diagnostics, environmental remediation, and agriculture, while also addressing the ethical and biosafety considerations surrounding its deployment. As CRISPR-Cas systems continue to evolve, they stand at the forefront of innovations that bridge natural microbial immunity with engineered precision tools for next-generation biotechnology.

The persistent global burden of cancer underscores the need for alternative therapeutic strategies beyond conventional chemotherapeutics, with metal-based agents offering distinct advantages due to their structural tunability and diverse biological modes of action. In this work, a series of thiosemicarbazone ligands bearing a progressively increasing number of aromatic rings were synthesized and employed to generate four Cu(I) complexes (1-4). Comprehensive characterization by spectroscopic techniques and single-crystal X-ray diffraction confirmed distorted tetrahedral geometries with bidentate N, S-coordination and stabilization by triphenylphosphine co-ligands. The cytotoxic potential of the ligands and complexes was evaluated against human cervical cancer (HeLa) cells using multiple in vitro assays. A clear structure-activity relationship was observed, wherein cytotoxic efficacy increased with aromatic ring density, with complex 4 exhibiting the strongest antiproliferative activity. Biochemical analyses revealed reduced glucose uptake, suppressed cell proliferation, and enhanced lactate dehydrogenase release. Apoptosis induction was validated by DAPI staining and Annexin V-FITC/PI flow cytometry, while in ovo chorioallantoic membrane assays demonstrated significant inhibition of neovascularization. Overall, these results establish ligand aromaticity as a key factor governing anticancer activity, providing insight into structure-activity correlations and highlighting the potential of Cu(I) complexes as promising candidates for further anticancer therapeutic development.

Fabrication and characterization of MTX-MWCNT nanoconjugate for evaluation of its cytotoxic activity, hemocompatibility, anti-angiogenic potential, molecular expression and morphological alterations through in vitro assays.

Urinary bladder cancer (UBCa) is a recurrent malignancy with a significant global health burden. Early and accurate detection is vital for improving outcomes. Metabolomics, the study of small molecules in biological systems, offers a powerful approach to reveal biochemical alterations in UBCa. Analysis of urine, tumor tissue, and blood highlights metabolic reprogramming, with mass spectrometry (MS) and nuclear magnetic resonance (NMR) as key profiling tools. Metabolites such as valine, tryptophan, taurine, uridine, and phosphocholine have emerged as potential biomarkers. Common alterations include reduced citrate and elevated lactic acid and fatty acids, reflecting dysregulated energy metabolism. The noninvasive nature of urine and blood sampling enhances clinical applicability. Overall, metabolomics provides valuable insights and a promising platform for early diagnosis and personalized therapy in UBCa.

Background: Endophytic fungi are known for diverse bioactive compounds with immense potential for agriculture and medicinal applications. Coniochaeta dendrobiicola isolated from root of the Dedrobium longicornu was investigated for its antioxidant and metabolite composition. The present study compares the antioxidant properties, flavonoid and phenolic contents and metabolic profiles of broth and mycelium extracts. Methanolic broth and mycelium extracts were tested for antioxidant potential using DPPH, while total flavonoid and phenolic content were measured using a UV-VIS spectrophotometer High-resolution mass spectrometry (HRMS) revealed a markedly richer and more diverse metabolite profile in the broth extract compared to the mycelium fraction. The broth extract exhibited significantly higher antioxidant activity, flavonoid, and phenolic contents, correlating with the presence of diverse bioactive compounds, including indole derivatives, flavonoids, phenolic acids, quinoline derivatives, and antifungal metabolites. Notably, several indole-related and phenolic compounds detected predominantly in the broth are known for antioxidant, antimicrobial, and plant growth–promoting properties. These findings indicate that C. dendrobiicola actively secretes biologically relevant secondary metabolites into the extracellular medium, highlighting its potential for agricultural and pharmaceutical applications.

The present study investigates the antibreast cancer potential of silver nanoparticles derived from the fungal endophyte Penicillium oxalicum (POAgNPs) using both in vitro and in vivo models. High-content analysis (HCA) demonstrated that POAgNPs significantly altered cellular and nuclear morphology, and we noted reduced cell impedance in human breast cancer cells MDA-MB-231 and MCF-7, indicating potent cytotoxicity. POAgNPs induced cell cycle arrest in the sub-G1 phase and promoted reactive oxygen species (ROS) generation, suggesting ROS-mediated apoptosis. Release studies confirmed efficient POAgNPs release at acidic pH (5.5), mimicking the tumor microenvironment, while the cellular uptake assay showed dose-dependent cytotoxicity. Moreover, POAgNPs exhibited antiangiogenic activity in the chick chorioallantoic membrane assay, indicating their potential to inhibit tumor neovascularization. In vivo, POAgNPs treatment mitigated the carcinogenic effects of 7,12-dimethylbenz[a]anthracene (DMBA) in breast tumor-bearing rats, as demonstrated by improved mammary tissue histology and significant restoration of oxygen saturation, total hemoglobin concentration, and reduced tumor vascularity measured via photoacoustic imaging (PAI). Biochemical assays revealed that POAgNPs helped restore the metabolic balance and oxidative status. Gene expression analysis showed upregulation of pro-apoptotic markers (CASPASE 8 and P53) and downregulation of antiapoptotic BCL-2 and the glucose transporter gene SLC2A1. Increased Caspase-3 activity further confirmed apoptosis induction. Collectively, these findings highlight the promise of POAgNPs as an effective antibreast cancer agent, warranting further research toward their development as a targeted nanotherapeutic.