CSIR-Indian Institute of Integrative Medicine

Cancer is a formidable adversary in contemporary healthcare. Routine screening and early diagnosis are crucial for favourable therapeutic outcomes. Publications, clinical trials, and patent landscape analysis suggest miRNA as promising biomarkers for diagnosis and prognosis of various cancers. This review intends to shed a holistic view of the current and futuristic methods for electrochemical biosensing platforms, using miRNA as biomarkers, coupled with microfluidics, machine learning techniques, and portable electronic devices. Electrochemical biosensors are thoroughly reviewed as they are promising candidate in the design and development of such devices where there is an in-depth exploration of the existing molecular techniques and sophisticated electrochemical biosensing strategies developed for the detection of miRNAs. Additionally, the review will critically analyze diverse signal enhancement strategies and microfluidic platforms specifically tailored for the detection of miRNA. Practical examples of such integrated electrochemical microfluidic biosensors are thoroughly cited along with the prospect of integration of these techniques with portable electronics, highlighting the future potential of highly integrated and accessible diagnostic solutions. Furthermore, the review will also encompass an assessment of the ongoing clinical trials investigating the utility of miRNA as cancer biomarker in diagnostic settings. Moreover, by assessing existing patents, the review shall provide a nuanced understanding of the intellectual property landscape, identifying key players, emerging technologies, and potential future directions. Our review with a 360-degree updated view on molecular biology components, electrochemical biosensors, engineering device design, clinical trials and patent landscape would appeal to researchers, engineers and clinicians working in the area of cancer molecular diagnosis.

Withania somnifera, commonly known as Ashwagandha, is widely utilized for treating neurological disorders in both traditional and modern medicine. While its neuroprotective effects are linked to several bioactive compounds, the individual safety profiles and potential cytotoxic effects of these compounds remain poorly understood. Previous studies from our research group have identified Withanone and Withanolide-A as neuroprotective agents. This study specifically evaluated the neurotoxic potential of Withaferin-A (WFA), a key steroidal lactone found in Withania somnifera, on differentiated SH-SY5Y cells. For six days, the SH-SY5Y cells were differentiated using 10 µM retinoic acid (RA). The neuronal phenotype was confirmed through morphological changes and increased expression of NeuN. Cytotoxicity assays demonstrated that WFA induces potent, dose-dependent cytotoxicity, resulting in approximately 50 %, 80 %, and 90 % cell death at concentrations of 0.6 µM, 1.2 µM, and 2.4 µM, respectively (p < 0.001). Treatment with WFA at 1.2 µM significantly increased both intracellular and mitochondrial reactive oxygen species (ROS), as shown by fluorescence imaging (p < 0.001). The loss of mitochondrial membrane potential was confirmed by JC-1 staining, indicating mitochondrial dysfunction. Western blot analysis indicated a dose-dependent increase in pro-apoptotic proteins Bax and Bid (p < 0.05 at 1.2 µM), an elevated Bax/Bcl-2 ratio (p < 0.001 at 1.2 µM), and enhanced activation of caspase-3, caspase-9, and cleavage of PARP-1 (p < 0.001 at 1.2 µM), indicating activation of the intrinsic apoptotic pathway. Molecular docking analysis revealed a strong binding affinity between WFA and PARP-1 (-8.2 kcal/mol), involving key residues Gly863, Ser904, and Tyr907, thereby supporting its role in PARP-1-mediated apoptosis. The computational findings were consistent with experimental observations of increased PARP-1 cleavage. This study concludes that WFA induces ROS-mediated mitochondrial dysfunction and caspase-dependent apoptosis in neuron-like cells. Consequently, future research should focus on critically assessing the safety and mechanistic effects of individual bioactive constituents of Withania somnifera, despite the well-established therapeutic potential of the plant.

The analogue of piperine 5-(3,4-methylenedioxyphenyl)-4-ethyl-2E,4E-pentadienoic acid piperidide (IIIM-1133), was evaluated in this in combination with pseudomonic acid A, commonly known as mupirocin against of S. aureus including Methicillin Resistant S. aureus (MRSA). Initial screening of compound revealed an increase in the potency of mupirocin, with a reduction in the minimum inhibitory concentration (MIC) values from 0.25 μg/mL to 0.06 μg/mL in case of wild type and MRSA strains of S. aureus and from 256 μg/mL to 32 μg/mL in mup^r-1 strain. The combination decreased the mutation frequency against mupirocin in wild-type S. aureus. The demonstrated ability of IIIM-1133 to suppress ethidium bromide efflux in MdeA-overexpressing S. aureus Mup^r-1 substantiated its effectiveness as an efflux pump inhibitor. Docking studies demonstrated that IIIM-1133 binds strongly in the substrate pocket of the MdeA efflux pump, highlighting its potential as an efflux pump inhibitor. IIIM-1133 alone and in combination with mupirocin was found to be nontoxic when tested on RAW 264.7 cells lines. A topical formulation of 2 % mupirocin with 0.5 % IIIM-1133 showed better efficacy in reducing the bacterial load in the infected site and sterilizing the infected patch. Furthermore, the formulation revealed better tissue healing potential with an anti-inflammatory activity by suppressing Il-6 and TNF-α. This formulation exhibited better efficacy than markedly available 2 % mupirocin.